WO1996017624A1 - Sunscreen-wound healing composition - Google Patents

Sunscreen-wound healing composition Download PDF

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Publication number
WO1996017624A1
WO1996017624A1 PCT/US1995/012848 US9512848W WO9617624A1 WO 1996017624 A1 WO1996017624 A1 WO 1996017624A1 US 9512848 W US9512848 W US 9512848W WO 9617624 A1 WO9617624 A1 WO 9617624A1
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WO
WIPO (PCT)
Prior art keywords
wound healing
acid
fatty acids
composition
sunscreen
Prior art date
Application number
PCT/US1995/012848
Other languages
French (fr)
Inventor
Alain Martin
Original Assignee
Warner-Lambert Company
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Warner-Lambert Company filed Critical Warner-Lambert Company
Priority to AU38596/95A priority Critical patent/AU690366B2/en
Priority to NZ295303A priority patent/NZ295303A/en
Priority to MX9703556A priority patent/MX9703556A/en
Priority to EP95936858A priority patent/EP0796107B1/en
Priority to DE69529345T priority patent/DE69529345T2/en
Publication of WO1996017624A1 publication Critical patent/WO1996017624A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/36—Carboxylic acids; Salts or anhydrides thereof
    • A61K8/361—Carboxylic acids having more than seven carbon atoms in an unbroken chain; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
    • A61K31/355—Tocopherols, e.g. vitamin E
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/35—Fat tissue; Adipocytes; Stromal cells; Connective tissues
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/36—Carboxylic acids; Salts or anhydrides thereof
    • A61K8/365—Hydroxycarboxylic acids; Ketocarboxylic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/16—Emollients or protectives, e.g. against radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/004—Aftersun preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/52—Stabilizers
    • A61K2800/522—Antioxidants; Radical scavengers
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S424/00—Drug, bio-affecting and body treating compositions
    • Y10S424/13—Burn treatment

Definitions

  • This invention pertains to therapeutic sunscreen-wound healing compositions useful to minimize and treat sunburn damage. More particularly, the sunscreen-wound healing compositions comprise a sunscreen agent, an anti-inflammatory agent, and a therapeutic wound healing composition and/or its metabolites. This invention also pertains to methods for preparing and using the therapeutic sunscreen-wound healing compositions and the pharmaceutical products in which the therapeutic compositions may be used.
  • a preferred embodiment of the therapeutic wound healing composition of this invention comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • Wounds are internal or external bodily injuries or lesions caused by physical means, such as mechanical, chemical viral, bacterial, or thermal means, which disrupt the normal continuity of structures.
  • Such bodily injuries include contusions, wounds in which the skin is unbroken, incisions, wounds in which the skin is broken by a cutting instrument, and lacerations, wounds in which the skin is broken by a dull or blunt instrument.
  • Wounds may be caused by accidents or by surgical procedures. Patients who suffer major wounds could benefit from an enhancement in the wound healing process.
  • Wound healing consists of a series of processes whereby injured tissue is repaired, specialized tissue is regenerated, and new tissue is reorganized. Wound healing consists of three major phases: a) an inflammation phase (0-3 days), b) a cellular proliferation phase (3-12 days), and (c) a remodeling phase (3 days-6 months).
  • platelet aggregation and clotting form a matrix which traps plasma proteins and blood cells to induce the influx of various types of ceils.
  • a wound healing step is desirable to resuscitate the injured cells and produce new cells to replace the dead cells.
  • the healing process requires the reversal of cytotoxicity, the suppression of inflammation, and the stimulation of cellular viability and proliferation.
  • Wounds require low levels of oxygen in the initial stages of healing to suppress oxidative damage and higher levels of oxygen in the later stages of healing to promote collagen formation by fibroblasts.
  • Mammalian cells are continuously exposed to activated oxygen species such as superoxide (O 2 -), hydrogen peroxide (H2O2), hydroxyl radical (OH ), and singlet oxygen ( O2).
  • these reactive oxygen intermediates are generated by cells in response to aerobic metabolism, catabolism of drugs and other xenobiotics, ultraviolet and x-ray radiation, and the respiratory burst of phagocytic cells (such as white blood cells) to kill invading bacteria such as those introduced through wounds.
  • Hydrogen peroxide for example, is produced during respiration of most living organisms especially by stressed and injured cells.
  • lipid peroxidation which involves the oxidative degradation of unsaturated lipids.
  • Lipid peroxidation is highly detrimental to membrane structure and function and can cause numerous cytopathological effects.
  • Cells defend against lipid peroxidation by producing radical scavengers such as superoxide dismutase, catalase, and peroxidase. Injured cells have a decreased ability to produce radical scavengers.
  • Hydrogen peroxide can react with DNA to cause backbone breakage, produce mutations, and alter and liberate bases. Hydrogen peroxide can also react with pyrimidines to open the 5, 6-double bond, which reaction inhibits the ability of pyrimidines to hydrogen bond to complementary bases, Hallaender et al. (1971). Such oxidative biochemical injury can result in the loss of cellular membrane integrity, reduced enzyme activity, changes in transport kinetics, changes in membrane lipid content, and leakage of potassium ions, amino acids, and other cellular material.
  • Antioxidants have been shown to inhibit damage associated with active oxygen species. For example, pyruvate and other ---//. ⁇ -ketoacids have been reported to react rapidly and stoichiometrically with hydrogen peroxide to protect cells from cytolytic effects, O'Donnell-Tormey et al., J. Exp. Med., 165, pp. 500-514 (1987).
  • United States Patents Nos. 3,920,835, 3,984,556, and 3,988,470 all issued to Van Scott et al., disclose methods for treating acne, dandruff, and palmar keratosis, respectively, which consist of applying to the affected area a topical composition comprising from about 1% to about 20% of a lower aliphatic compound containing from two to six carbon atoms selected from the group consisting of Alpha- hydroxyacids, -*-/ . ⁇ -ketoacids and esters thereof, and 3-hydroxybutryic acid in a pha ⁇ naceutically acceptable carrier.
  • the aliphatic compounds include pyruvic acid and lactic acid.
  • United States Patents Nos. 4,105,783 and 4,197,316 both issued to Yu et al., disclose a method and composition, respectively, for treating dry skin which consists of applying to the affected area a topical composition comprising from about 1 % to about 20% of a compound selected from the group consisting of amides and ammonium salts of Alp A ⁇ -hydroxyacids, jS-hydroxyacids, and -4//> ⁇ -ketoacids in a pharmaceutically acceptable carrier.
  • the compounds include the amides and ammonium salts of pyruvic acid and lactic acid.
  • United States Patent No. 4,234,599 issued to Van Scott et al., discloses a method for treating actinic and nonactinic skin keratoses which consists of applying to the affected area a topical composition comprising an effective amount of a compound selected from the group consisting of -4//. ⁇ -hydroxyacids, jS-hydroxyacids, and -4//. ⁇ -.-ketoacids in a pharmaceutically acceptable carrier.
  • the acidic compounds include pyruvic acid and lactic acid.
  • United States Patent No. 4,294,852 issued to Wildnauer et al., discloses a composition for treating skin which comprises the Alpha-hydroxyacids. ⁇ -hydroxyacids, and /p ⁇ -ketoacids disclosed above by Van Scott et al. in combination with C ⁇ -Cg aliphatic alcohols.
  • United States Patent No. 4,663,166 issued to Veech, discloses an electrolyte solution which comprises a mixture of L-lactate and pyruvate in a ratio from 20: 1 to 1:1, respectively, or a mixture of D- ⁇ -hydroxybutyrate and acetoacetate, in a ratio from 6:1 to 0.5:1, respectively.
  • Sodium pyruvate has been reported to reduce the number of erosions, ulcers, and hemorrhages on the gastric mucosa in guinea pigs and rats caused by acetylsalicylic acid.
  • the analgesic and antipyretic properties of acetylsalicylic acid were not impaired by sodium pyruvate, Puschmann, Arzneistoffforschung, 33, pp. 410-415 and 415-416 (1983).
  • Pyruvate has been reported to produce a relative stabilization of left ventricular pressure and work parameter and to reduce the size of infarctions. Pyruvate improves resumption of spontaneous beating of the heart and restoration of normal rates and pressure development, Bunger et al, J. Mol. Cell. Cardiol., 18, pp. 423*438 (1986), Mochizuki et al, J. Physiol. (Paris), 76, pp. 805-812 (1980), Regitz et al, Cardiovasc. Res., 15, pp. 652-658 (1981), Giannelli et al, Ann. Thorac. Surg., 21, pp. 386-396 (1976).
  • Sodium pyruvate has been reported to act as an antagonist to cyanide intoxication (presumably through the formation of a cyanohydrin) and to protect against the lethal effects of sodium sulfide and to retard the onset and development of functional, morphological, and biochemical measures of acrylamide neuropathy of axons, Schwartz et ⁇ /., Toxicol. Appl. Pharmacol., 50, pp. 437-442 (1979), Sabri et al, Brain Res., 483, pp. 1-11 (1989).
  • United States Patents Nos. 4,158,057, 4,351,835, 4,415,576, and 4,645,764, all issued to Stanko disclose methods for preventing the accumulation of fat in the liver of a mammal due to the ingestion of alcohol, for controlling weight in a mammal, for inhibiting body fat while increasing protein concentration in a mammal, and for controlling the deposition of body fat in a living being, respectively.
  • the methods comprise administering to the mammal a therapeutic mixture of pyruvate and dihydroxyacetone, and optionally riboflavin.
  • 4,548,937 discloses a method for controlling the weight gain of a mammal which comprises administering to the mammal a therapeutically effective amount of pyruvate, and optionally riboflavin.
  • United States Patent No. 4,812,479, issued to Stanko discloses a method for controlling the weight gain of a mammal which comprises administering to the mammal a therapeutically effective amount of dihydroxyacetone, and optionally riboflavin and pyruvate.
  • Rats fed a calcium-oxalate lithogenic diet including sodium pyruvate were reported to develop fewer urinary calculi (stones) than control rats not given sodium pyruvate, Ogawa et al, Hinyokika Kiyo, 32, pp. 1341-1347 (1986).
  • United States Patent No. 4,521,375 discloses a method for sterilizing surfaces which come into contact with living tissue. The method comprises sterilizing the surface with aqueous hydrogen peroxide and then neutralizing the surface with pyruvic acid.
  • United States Patent No. 4,416,982, issued to Tauda et al discloses a method for decomposing hydrogen peroxide by reacting the hydrogen peroxide with a phenol or aniline derivative in the presence of peroxidase.
  • United States Patent No. 4,696,917 discloses an eye irrigation solution which comprises Eagle's Minimum Essential Medium with Earle's salts, chondroitin sulfate, a buffer solution, 2-mercaptoethanol, and a pyruvate.
  • the irrigation solution may optionally contain ascorbic acid and ---/p ⁇ -tocopherol.
  • United States Patent No. 4,725,586, issued to Lindstrom et al discloses an irrigation solution which comprises a balanced salt solution, chondroitin sulfate, a buffer solution,
  • the irrigation solution may optionally contain ascorbic acid mXnd mma-xocc herol.
  • United States Patent No. 4,847,069 discloses a photoprotective composition comprising (a) a sorbohydroxamic acid, (b) an anti- inflammatory agent selected from steroidal anti-inflammatory agents and a natural anti- inflammatory agent, and (c) a topical carrier. Fatty acids may be present as an emollient.
  • United States Patent No. 4,847,071 issued to Bissett et al, discloses a photoprotective composition comprising (a) a tocopherol or tocopherol ester radical scavenger, (b) an anti-inflammatory agent selected from steroidal anti-inflammatory agents and a natural anti-inflammatory agent, and (c) a topical carrier.
  • United States Patent No. 4,847,072, issued to Bissett et al discloses a topical composition comprising not more than 25% tocopherol sorbate in a topical carrier.
  • United States Patent No. 4,533,637 issued to Yamane et al, discloses a culture medium which comprises a carbon source, a nucleic acid source precursor, amino acids, vitamins, minerals, a lipophilic nutrient, and serum albumin, and cyclodextrins.
  • the lipophilic substances include unsaturated fatty acids and lipophilic vitamins such as Vitamin A, D, and E. Ascorbic acid may also be present.
  • United Kingdom patent application no. 2,196,348A discloses a synthetic culture medium which comprises inorganic salts, monosaccharides, amino acids, vitamins, buffering agents, and optionally sodium pyruvate adding magnesium hydroxide or magnesium oxide to the emulsion.
  • the oil phase may include chicken fat.
  • United States Patent No. 4,284,630 issued to Yu et al, discloses a method for stabilizing a water-in-oil emulsion which comprises adding magnesium hydroxide or magnesium oxide to the emulsion.
  • the oil phase may include chicken fat.
  • Preparation H has been reported to increase the rate of wound healing in artificially created rectal ulcers.
  • the active ingredients in Preparation H are skin respiratory factor and shark liver oil, Subramanyam et al, Digestive Diseases and Sciences, 29, pp. 829-832 (1984).
  • European patent application no. 0410696A1 discloses a mucoadhesive delivery system comprising a treating agent and a polyacrylic acid cross- linked with from about 1% to about 20% by weight of a polyhydroxy compound such as a sugar, cyclitol, or lower polyhydric alcohol.
  • Inflammation is a nonspecific response caused by several kinds of injury, including penetration of the host by infectious agents.
  • the distinguishing feature of inflammation is dilation and increased permeability of minute blood vessels.
  • Direct injury such as that caused by toxins elaborated by microorganisms, leads to destruction of vascular endothelium and increased permeability to plasma proteins, especially in the venules and venular capillaries.
  • Mediators of secondary injury are liberated from the site of direct injury. As a result, gaps form between vascular endothelial cells through which plasma proteins escape. Granulocytes, monocytes, and erythrocytes may also leave vascular channels.
  • Mediators of secondary injury include unknown substances and histamine, peptides (kinins), kinin-forming enzymes (kininogenases), and a globulin permeability factor. These mediators are blocked from action by antihistamines and sympathoamines, and are most pronounced in effect on venules, although lymphvascular endothelium also becomes more porous as a part of secondary injury.
  • the beneficial effect of the inflammatory response is the production of: (1) leukocytes in great numbers; (2) plasma proteins, nonspecific and specific humoral agents, fibrinogen that on conversion to fibrin aids in localization of the infectious process while acting as a matrix for phagocytosis; and (3) increased blood and lymph flow that dilutes and flushes toxic materials while causing a local increase in temperature.
  • the exudate In the early stages of inflammation, the exudate is alkaline and neutrophilic polymorphonuclear leukocytes predominate. As lactic acid accumulates, presumably from glycolysis, the Ph drops and macrophages become the predominant cell type.
  • Lactic acid and antibodies in the inflammatory exudate may inhibit parasites, but the major anti-infectious effect of the inflammatory response is attributable to phagocytic cells.
  • the inflammatory response consists of three successive phases: (a) increased vascular permeability with resulting edema and swelling, (b) cellular infiltration and phagocytoses, and (c) proliferation of the fibroblasts, synthesizing new connective tissue to repair the injury.
  • mediators of inflammation have been implicated in the inflammatory process primarily in terms of their capacity to induce vasodilation and increase permeability.
  • the initial increase in capillary permeability and vasodilation in an inflamed joint is followed by an increase in metabolism of the joint tissues.
  • Leakage of fibrinogen into the wound, where proteolytic enzymes convert it into fibrin establishes a capillary and lymphatic blockade.
  • the concentrations of components of the ground substance of connective tissue collagen, mucopolysaccharides, glycoproteins, and nonfibrous proteins are greatly increased during this process.
  • the fibroblast is found to be the dominant cell in the wounded zone. It first proliferates, then synthesizes extracellular material, including new collagen fibers and acid mucopolysaccharides, which are laid down to form the new tissue matrix.
  • the inflammatory phenomenon includes fenestration of the microvasculature, leakage of the elements of blood into the interstitial spaces, and migration of leukocytes into the inflamed tissue. On a macroscopic level, this is usually accompanied by the familiar clinical signs of erythema, edema tenderness
  • Sunburn is an acute reaction by the skin to excessive exposure to sunlight. Ordinary sunburn results from overexposure of the skin to ultraviolet waves of about 3000 Angstroms. Sunburn symptoms appear in 1 to 24 hours and, except in severe reactions, pass their peak in 72 hours. Following exposure to sunlight, the epidermis thickens and the melanocytes produce melanin at an increased rate, providing some natural protection against further exposure. Skin changes range from a mild erythema with subsequent evanescent scaling, to pain, swelling, skin tenderness, and blisters from more prolonged exposure. Fever, chills, weakness, and shock may appear if a large portion of the body surface is affected. Secondary infection, particularly furunculosis, is the most common late complication. The skin may remain hypervulnerable to sunlight for one to several weeks when pronounced exfoliation has occurred.
  • Sunscreen agents are very effective for preventing sunburn.
  • a useful sunscreen agent formulation is 5% . ⁇ r -aminobenzoic acid (PABA) in ethyl alcohol or in a gel.
  • PABA 5% . ⁇ r -aminobenzoic acid
  • the esters of ⁇ r ⁇ -aminobenzoic acid in an alcohol base are only slightly less effective. Patients who do not tolerate ⁇ ra-aminobenzoic acid or its esters may use a benzophenone or titanium dioxide sunscreen agent.
  • Early treatment of extensive and severe sunburn with a topical or systemic corticosteroid decreases discomfort considerably.
  • compositions of this invention comprise a therapeutically effective amount of (1) a sunscreen agent; (2) an anti- inflammatory agent; and, (3) a wound healing composition.
  • a preferred embodiment of the wound healing composition of this invention comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the therapeutic sunscreen-wound healing compositions of this invention may be utilized in a wide variety of pharmaceutical products. This invention also relates to methods for preparing and using the therapeutic sunscreen-wound healing compositions and the pharmaceutical products in which the therapeutic compositions may be used.
  • This invention further comprises augmented therapeutic sunscreen-wound healing compositions comprising (1) a sunscreen agent; (2) an anti-inflammatory agent; and, (3) a therapeutic wound healing composition in combination with one or more additional medicaments.
  • This invention also relates to methods for preparing and using the augmented therapeutic antikeratolytic-wound healing compositions and the pharmaceutical products in which the augmented compositions may be used.
  • Figure 1 depicts in bar graph format the viability of U937 monocytic cells following exposure of the cells to various antioxidants (Examples 1-5).
  • Figure 2 depicts in bar graph format the viability of U937 monocytic cells following exposure of the cells to various combinations of antioxidants (Examples 6- 13).
  • Figure 3 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells following exposure of the cells to various antioxidants (Examples 14-18).
  • Figure 4 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells following exposure of the cells to various combinations of antioxidants (Examples 19-26).
  • Figure 5 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells following exposure of the cells to various combinations of antioxidants with and without a mixture of saturated and unsaturated fatty acids (Examples 27-32).
  • Figure 6 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal keratinocytes following exposure of the cells to various antioxidants with and without a mixture of saturated and unsaturated fatty acids (Examples 33-42).
  • Figure 7 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal keratinocytes following exposure of the cells to various combinations of antioxidants with and without a mixture of saturated and unsaturated fatty acids (Examples 43-52).
  • Figure 8 depicts in bar graph format a summary analysis of the levels of hydrogen peroxide produced by epidermal keratinocytes following exposure of the cells to the individual components of the wound healing composition, to various combinations of the wound healing composition, and to the wound healing composition.
  • Figure 9 is a photograph of wounded mice after 4 days of treatment with:
  • Preparation H (Example A); a petrolatum base formulation containing live yeast cell derivative, shark oil, and a mixture of sodium pyruvate, vitamin E, and chicken fat (Example B); a petrolatum base formulation containing live yeast cell derivative and shark oil (Example C); and no composition (Example E, control).
  • Figure 10 is a photograph of a wounded mouse after 4 days of treatment with a petrolatum base formulation only (Example D).
  • This invention pertains to therapeutic sunscreen-wound healing compositions which comprise (1) a sunscreen agent; (2) an anti-inflammatory; and, (3) a wound healing composition and/or its metabolites.
  • the wound healing composition comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • Applicant has discovered therapeutic wound healing compositions for preventing and reducing injury to mammalian cells and increasing the resuscitation rate of injured mammalian cells.
  • Cells treated with the therapeutic wound healing compositions of the present invention show decreased levels of hydrogen peroxide production, increased resistance to cytotoxic agents, increased rates of proliferation, and increased viability.
  • Cellular cultures containing the therapeutic wound healing compositions showed enhanced differentiation and proliferation over control cultures and rapidly formed attachments or tight junctions between the cells to form an epidermal sheet.
  • Wounded mammals treated with the therapeutic wound healing compositions show significantly improved wound closing and healing over untreated mammals and mammals treated with conventional healing compositions.
  • the wound healing compositions may be used alone or in combination with other medicaments.
  • the therapeutic wound healing compositions of this invention are described as Embodiment One.
  • the therapeutic wound healing composition comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the therapeutic wound healing composition comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the therapeutic wound healing composition comprises (a) an antioxidant and (b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the therapeutic wound healing composition comprises (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the therapeutic wound healing compositions of this invention are further combined with a therapeutically effective amount of (1) a sunscreen agent and (2) an anti-inflammatory (collectively referred to as "X") to form therapeutic sunscreen-wound healing compositions (I.A-D + X).
  • the therapeutic sunscreen-wound healing compositions may be used alone or in combination with other medicaments.
  • This invention also pertains to methods for preparing and using the sunscreen- wound healing compositions and the pharmaceutical products in which the therapeutic compositions may be used.
  • the therapeutic sunscreen-wound healing compositions of this invention may be further combined with one or more additional medicaments for treating wounds to form augmented sunscreen-wound healing compositions.
  • This invention also relates to methods for preparing and using the augmented therapeutic sunscreen-wound healing compositions and the pharmaceutical products in which the augmented compositions may be used.
  • an injured cell means a cell that has any activity disrupted for any reason.
  • an injured cell may be a cell that has injured membranes or damaged DNA, RNA, and ribosomes, for example, a cell which has (a) injured membranes so that transport through the membranes is diminished resulting in an increase in toxins and normal cellular wastes inside the cell and a decrease in nutrients and other components necessary for cellular repair inside the cell, (b) an increase in concentration of oxygen radicals inside the cell because of the decreased ability of the cell to produce antioxidants and enzymes, or (c) damaged DNA, RNA, and ribosomes which must be repaired or replaced before normal cellular functions can be resumed.
  • resuscitation of injured mammalian cells means the reversal of cytotoxicity, the stabilization of the cellular membrane, an increase in the proliferation rate of the cell, and or the normalization of cellular functions such as the secretion of growth factors, hormones, and the like.
  • cytotoxicity as used herein means a condition caused by a cytotoxic agent that injures the cell. Injured cells do not proliferate because injured cells expend all energy on cellular repair. Aiding cellular repair promotes cellular proliferation.
  • prodrug refers to compounds which undergo biotransformation prior to exhibiting their pharmacological effects.
  • drug latentiation is the chemical modification of a biologically active compound to form a new compound which upon in vivo enzymatic attack will liberate the parent compound.
  • the chemical alterations of the parent compound are such that the change in physicochemical properties will affect the absorption, distribution and enzymatic metabolism.
  • the definition of drug latentiation has also been extended to include nonenzymatic regeneration of the parent compound. Regeneration takes place as a consequence of hydrolytic, dissociative, and other reactions not necessarily enzyme mediated.
  • prodrugs latentiated drugs, and bioreversible derivatives are used interchangeably.
  • latentiation implies a time lag element or time component involved in regenerating the bioactive parent molecule in vivo.
  • prodrug is general in that it includes latentiated drug derivatives as well as those substances which are converted after administration to the actual substance which combines with receptors.
  • prodrug is a generic term for agents which undergo biotransformation prior to exhibiting their pharmacological actions.
  • the term "prodrug” also includes compounds which may not necessarily undergo biotransformation to the administered drug but may undergo biotransformation to the active agent which exhibits the desired pharmacological effect.
  • metabolic refers to any substance produced by metabolism or by a metabolic process.
  • Metabolism refers to the various chemical reactions involved in the transformation of molecules or chemical compounds occurring in tissue and the cells therein.
  • the cells which may be treated with the therapeutic wound healing compositions in the present invention are mammalian cells. Although applicant will describe the present therapeutic wound healing compositions as useful for treating mammalian epidermal keratinocytes and mammalian monocytes, applicant contemplates that the therapeutic wound healing compositions may be used to protect or resuscitate all mammalian cells. Keratinocytes are representative of normal mammalian cells and are the fastest proliferating cells in the body. The correlation between the reaction of keratinocytes to injury and therapy and that of mammalian cells in general is very high. Monocytes are representative of specialized mammalian cells such as the white blood cells in the immune system and the organ cells in liver, kidney, heart, and brain. The mammalian cells may be treated in vivo and in vitro.
  • Epidermal keratinocytes are the specialized epithelial cells of the epidermis which synthesize keratin, a scleroprotein which is the principal constituent of epidermis, hair, nails, homy tissue, and the organic matrix of the enamel of teeth.
  • Mammalian epidermal keratinocytes constitute about 95% of the epidermal cells and together with melanocytes form the binary system of the epidermis. In its various successive stages, epidermal keratinocytes are also known as basal cells, prickle cells, and granular cells.
  • Monocytes are mononuclear phagocytic leukocytes which undergo respiratory bursting and are involved in reactive oxygen mediated damage within the epidermis.
  • Leukocytes are white blood cells or corpuscles which may be classified into two main groups: granular leukocytes (granulocytes) which are leukocytes with abundant granules in the cytoplasm and nongranular leukocytes (nongranulocytes) which are leukocytes without specific granules in the cytoplasm and which include the lymphocytes and monocytes.
  • Phagocyte cells are cells which ingest microorganisms or other cells and foreign particles. Monocytes are also known as large mononuclear leukocytes, and hyaline or transitional leukocytes.
  • Epidermal keratinocytic cells and monocytic cells have multiple oxygen generating mechanisms and the degree to which each type of mechanism functions differs in each type of cell.
  • monocytes for example, the respiratory bursting process is more pronounced than in epidermal keratinocytes.
  • the components in the therapeutic wound healing compositions of the present invention may vary depending upon the types of cells involved in the condition being treated.
  • the therapeutic wound healing composition for treating mammalian cells preferably epidermal keratinocytes, comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the therapeutic wound healing composition for treating mammalian cells comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the therapeutic wound healing composition for treating mammalian cells comprises (a) an antioxidant and (b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the therapeutic wound healing composition for treating mammalian cells preferably monocytes, comprises (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • Pyruvic acid (2-oxopropanoic acid ⁇ /pA -ketopropionic acid, CH3COCOOH) or pyruvate is a fundamental intermediate in protein and carbohydrate metabolism and in the citric acid cycle.
  • the citric acid cycle (tricarboxylic acid cycle, Kreb's cycle) is the major reaction sequence which executes the reduction of oxygen to generate adenosine triphosphate (ATP) by oxidizing organic compounds in respiring tissues to provide electrons to the transport system.
  • Acetyl coenzyme A (“active acetyl”) is oxidized in this process and is thereafter utilized in a variety of biological processes and is a precursor in the biosynthesis of many fatty acids and sterols.
  • acetyl coenzyme A The two major sources of acetyl coenzyme A are derived from the metabolism of glucose and fatty acids. Glycolysis consists of a series of transformations wherein each glucose molecule is transformed in the cellular cytoplasm into two molecules of pyruvic acid. Pyruvic acid may then enter the mitochondria where it is oxidized by coenzyme A in the presence of enzymes and cofactors to acetyl coenzyme A. Acetyl coenzyme A can then enter the citric acid cycle.
  • pyruvic acid derived from glycogen
  • Lactic acid is reoxidized and partially retransformed to glycogen during rest.
  • Pyruvate can also act as an antioxidant to neutralize oxygen radicals in the cell and can be used in the multifunction oxidase system to reverse cytotoxicity.
  • the pyruvate in the present invention may be selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, prodrugs of pyruvic acid, and mixtures thereof.
  • the pharmaceutically acceptable salts of pyruvic acid may be alkali salts and alkaline earth salts.
  • the pyruvate is selected from the group consisting of pyruvic acid, lithium pyruvate, sodium pyruvate, potassium pyruvate, magnesium pyruvate, calcium pyruvate, zinc pyruvate, manganese pyruvate, methyl pyruvate, Alpha-ketoglutaric acid, and mixtures thereof.
  • the pyruvate is selected from the group of salts consisting of sodium pyruvate, potassium pyruvate, magnesium pyruvate, calcium pyruvate, zinc pyruvate, manganese pyruvate, and the like, and mixtures thereof. Most preferably, the pyruvate is sodium pyruvate.
  • the amount of pyruvate present in the therapeutic wound healing compositions of the present invention is a therapeutically effective amount.
  • a therapeutically effective amount of pyruvate is that amount of pyruvate necessary for the inventive composition to prevent and reduce injury to mammalian cells or increase the resuscitation rate of injured mammalian cells.
  • the exact amount of pyruvate is a matter of preference subject to such factors as the type of condition being treated as well as the other ingredients in the composition.
  • pyruvate is present in the therapeutic wound healing composition in an amount from about 10% to about 50%, preferably from about 20% to about 45%, and more preferably from about 25% to about 40%, by weight of the therapeutic wound healing composition.
  • lactate occurs in small quantities in the blood and muscle fluid of mammals. Lactic acid concentration increases in muscle and blood after vigorous activity. Lactate is a component in the cellular feedback mechanism and inhibits the natural respiratory bursting process of cells thereby suppressing the production of oxygen radicals.
  • the lactate in the present invention may be selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, prodrugs of lactic acid, and mixtures thereof.
  • the pharmaceutically acceptable salts of lactic acid may be alkali salts and alkaline earth salts.
  • the lactate is selected from the group consisting of lactic acid, lithium lactate, sodium lactate, potassium lactate, magnesium lactate, calcium lactate, zinc lactate, manganese lactate, and the like, and mixtures thereof. More preferably, the lactate is selected from the group consisting of lactic acid, sodium lactate, potassium lactate, magnesium lactate, calcium lactate, zinc lactate, manganese lactate, and mixtures thereof. Most preferably, the lactate is lactic acid.
  • the amount of lactate present in the therapeutic wound healing compositions of the present invention is a therapeutically effective amount.
  • a therapeutically effective amount of lactate is that amount of lactate necessary for the inventive composition to prevent and reduce injury to mammalian cells or increase the resuscitation rate of injured mammalian cells.
  • a therapeutically effective amount of lactate is that amount necessary to suppress the respiratory bursting process of white blood cells to protect and resuscitate the mammalian cells.
  • a therapeutically effective amount of lactate in an ingestible composition is from about 5 to about 10 times the amount of lactate normally found in serum. The exact amount of lactate is a matter of preference subject to such factors as the type of condition being treated as well as the other ingredients in the composition.
  • lactate is present in the therapeutic wound healing composition in an amount from about 10% to about 50%, preferably from about 20% to about 45%, and more preferably from about 25% to about 40%, by weight of the therapeutic wound healing composition.
  • Antioxidants are substances which inhibit oxidation or suppress reactions promoted by oxygen or peroxides. Antioxidants, especially lipid-soluble antioxidants, can be absorbed into the cellular membrane to neutralize oxygen radicals and thereby protect the membrane.
  • the antioxidants useful in the present invention may be selected from the group consisting of all forms of Vitamin A (retinol), all forms of
  • Vitamin2 (3, 4-didehydroretinol), all forms of carotene such as ---//. A ⁇ -carotene, ⁇ - carotene (beta, .-carotene), g ⁇ / ⁇ m ⁇ -carotene, de/t ⁇ -carotene, all forms of Vitamin C (D-ascorbic acid, L-ascorbic acid), all forms of tocopherol such as Vitamin E (Alpha- tocopherol, 3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltri-decyl)-2H-l- benzopyran-6-ol), /.-tocopherol, g ⁇ mm ⁇ -tocopherol, rfe/t ⁇ -tocopherol, tocoquinone, tocotrienol, and Vitamin E esters which readily undergo hydrolysis to Vitamin E such as Vitamin E acetate and Vitamin E succinate, and pharmaceutically acceptable Vitamin E salts such as Vitamin E phosphat
  • the antioxidant is selected from the group of lipid-soluble antioxidants consisting of Vitamin A, j ⁇ -carotene, Vitamin E, Vitamin E acetate, and mixtures thereof. More preferably, the antioxidant is Vitamin E or Vitamin E acetate. Most preferably, the antioxidant is Vitamin E acetate.
  • the amount of antioxidant present in the therapeutic wound healing compositions of the present invention is a therapeutically effective amount.
  • a therapeutically effective amount of antioxidant is that amount of antioxidant necessary for the inventive composition to prevent and reduce injury to mammalian cells or increase the resuscitation rate of injured mammalian cells.
  • the exact amount of antioxidant is a matter of preference subject to such factors as the type of condition being treated as well as the other ingredients in the composition.
  • the antioxidant is present in the therapeutic wound healing composition in an amount from about 0.1% to about 40%, preferably from about 0.2% to about 30%, and more preferably from about 0.5% to about 20%, by weight of the therapeutic wound healing composition.
  • the mixture of saturated and unsaturated fatty acids in the present invention are those fatty acids required for the repair of mammalian cellular membranes and the production of new cells.
  • Fatty acids are carboxylic acid compounds found in animal and vegetable fat and oil.
  • Fatty acids are classified as lipids and are composed of chains of alkyl groups containing from 4 to 22 carbon atoms and 0-3 double bonds and characterized by a terminal carboxyl group, -COOH.
  • Fatty acids may be saturated or unsaturated and may be solid, semisolid, or liquid.
  • the most common saturated fatty acids are butyric acid (C4), lauric acid (C ⁇ ), palmitic acid (C j g), and stearic acid (C j g).
  • Unsaturated fatty acids are usually derived from vegetables and consist of alkyl chains containing from 16 to 22 carbon atoms and 0-3 double bonds with the characteristic terminal carboxyl group.
  • the most common unsaturated fatty acids are oleic acid, linoleic acid, and linolenic acid (all C j o acids).
  • the mixture of saturated and unsaturated fatty acids required for the repair of mammalian cellular membranes in the present invention may be derived from animal and vegetable fats and waxes, prodrugs of saturated and unsaturated fatty acids useful in the present invention, and mixtures thereof.
  • the fatty acids in the therapeutic wound healing composition may be in the form of mono-, di-, or trigylcerides, or free fatty acids, or mixtures thereof, which are readily available for the repair of injured cells.
  • Cells produce the chemical components and the energy required for cellular viability and store excess energy in the form of fat. Fat is adipose tissue stored between organs of the body to furnish a reserve supply of energy.
  • the preferred animal fats and waxes have a fatty acid composition similar to that of human fat and the fat contained in human breast milk.
  • the prefe ⁇ ed animal fats and waxes may be selected from the group consisting of human fat, chicken fat, cow fat (defined herein as a bovine domestic animal regardless of sex or age), sheep fat, horse fat, pig fat, and whale fat.
  • the more prefe ⁇ ed animal fats and waxes may be selected from the group consisting of human fat and chicken fat. The most prefe ⁇ ed animal fat is human fat.
  • oils especially sunflower oil
  • marine oils especially shark liver oil
  • synthetic waxes and oils which have a fatty acid composition similar to that of animal fats and waxes, and preferably to that of human fats and waxes, may also be employed.
  • the mixture of saturated and unsaturated fatty acids has a composition similar to that of human fat and comprises the following fatty acids: butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, myristoleic acid, palmitic acid, paimitoleic acid, stearic, oleic acid, linoleic acid, linolenic acid, arachidic acid, and gadoleic acid.
  • butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, myristoleic acid, palmitic acid, paimitoleic acid, stearic, oleic acid, linoleic acid, linolenic acid, arachidic acid, and gadoleic acid are present in the mixture in about the following percentages by weight, respectively (carbon chain number and number of unsaturations are shown parenthetically, respectively): 0.2%-0.4% (C 4 ), 0.1% (C 6 ), 0.3%-0.8% (Co), 2.2%-3.5% (C 10 ), 0.9%-5.5% (C 12 ), 2.8%-8.5% (C 14 ), 0.1%-0.6% (C 14: 1 ), 23.2%-24.6% (C J 6 ), 1.8%-3.0% (C 16; 1 ), 6.9%-9.9% (C ] 8 ), 36.0%-36.5% (C l g: 1 ), 20%-20.6% (C 18:2
  • the mixture of saturated and unsaturated fatty acids is typically chicken fat comprising the following fatty acids: lauric acid, myristic acid, myristoleic acid, pentadecanoic acid, palmitic acid, paimitoleic acid, margaric acid, margaroleic acid, stearic, oleic acid, linoleic acid, linolenic acid, arachidic acid, and gadoleic acid.
  • lauric acid, myristic acid, myristoleic acid, pentadecanoic acid, palmitic acid, paimitoleic acid, margaric acid, margaroleic acid, stearic, oleic acid, linoleic acid, linolenic acid, arachidic acid, and gadoleic acid are present in the mixture in about the following percentages by weight, respectively: 0.1% (C 12 ), 0.8% (C 14 ), 0.2% (C 14: ] ), 0.1% (C 15 ), 25.3% (C 16 ), 7.2% (C 16: 1 ), 0.1% (C ), 0.1% (C 17 . j ), 6.5% (C l g ), 37.7% (C l g: 1 ), 20.6% (C 18:2 ), 0.8% (C 18;3 ), 0.2%
  • the mixture of saturated and unsaturated fatty acids comprises lecithin.
  • Lecithin phosphatidylcholine
  • Lecithin is a phosphatide found in all living organisms (plants and animals) and is a significant constituent of nervous tissue and brain substance.
  • Lecithin is a mixture of the diglycerides of stearic, palmitic, and oleic acids, linked to the choline ester of phosphoric acid.
  • the product of commerce is predominantly soybean lecithin obtained as a by-product in the manufacturing of soybean oil.
  • Soybean lecithin contains palmitic acid 11.7%, stearic 4.0%, paimitoleic 8.6%, oleic 9.8%, linoleic 55.0%, linolenic 4.0%, C 0 to C 2 acids (includes arachidonic) 5.5%.
  • Lecithin may be represented by the fo ⁇ nula:
  • R is selected from the group consisting of stearic, palmitic, and oleic acid.
  • fatty acids and percentages thereof present in the fatty acid mixture are given as an example.
  • the exact type of fatty acid present in the fatty acid mixture and the exact amount of fatty acid employed in the fatty acid mixture may be varied in order to obtain the result desired in the final product and such variations are now within the capabilities of those skilled in the art without the need for undue experimentation.
  • the amount of fatty acids present in the therapeutic wound healing compositions of the present invention is a therapeutically effective amount.
  • a therapeutically effective amount of fatty acids is that amount of fatty acids necessary for the inventive composition to prevent and reduce injury to mammalian cells or increase the resuscitation rate of injured mammalian cells.
  • the exact amount of fatty acids employed is subject to such factors as the type and distribution of fatty acids employed in the mixture, the type of condition being treated, and the other ingredients in the composition.
  • the fatty acids are present in the therapeutic wound healing composition in an amount from about 10% to about 50%, preferably from about 20% to about 45%, and more preferably from about 25% to about 40%, by weight of the therapeutic wound healing composition.
  • the therapeutic wound healing compositions of Embodiment One (I.A-D) for treating mammalian cells may be selected from the group consisting of:
  • (I.A)(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • (I.D) (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; (b) an antioxidant; and
  • fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the wound healing compositions of Embodiment One (I) for treating mammalian cells may be selected from the group consisting of:
  • LA (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • LB (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof;
  • (I) for treating mammalian cells may be selected from the group consisting of: (LA) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • the wound healing compositions of Embodiment One (I) for treating mammalian cells may be selected from the group consisting of:
  • LA (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • LB (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof;
  • the wourd healing compositions of Embodiment One (I) for treating mammalian cells preferably epidermal keratinocytes, comprise:
  • LA (a) pyruvate selected from the group consisting of pyruvic acid, pha ⁇ naceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the wound healing compositions of Embodiment One (I) for treating mammalian cells preferably monocytes, comprise:
  • LD lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof;
  • fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • pyruvate can be transported inside a cell where it can act as an antioxidant to neutralize oxygen radicals in the cell.
  • Pyruvate can also be used inside the cell in the citric acid cycle to provide energy to increase cellular viability, and as a precursor in the synthesis of important biomolecules to promote cellular proliferation.
  • pyruvate can be used in the multifunction oxidase system to reverse cytotoxicity.
  • Antioxidants, especially lipid-soluble antioxidants can be absorbed into the cell membrane to neutralize oxygen radicals and thereby protect the membrane.
  • the saturated and unsaturated fatty acids in the present invention are those fatty acids required for the resuscitation of mammalian cells and are readily available for the repair of injured cells and the proliferation of new cells.
  • Cells injured by oxygen radicals need to produce unsaturated fatty acids to repair cellular membranes.
  • the production of unsaturated fatty acids by cells requires oxygen.
  • the injured cell needs high levels of oxygen to produce unsaturated fatty acids and at the same time needs to reduce the level of oxygen within the cell to reduce oxidative injury.
  • the need of the cell for unsaturated fatty acids is reduced and the need for high oxygen levels is also reduced.
  • the combination of pyruvate inside the cell and an antioxidant in the cellular membrane functions in an unexpected synergistic manner to reduce hydrogen peroxide production in the cell to levels lower than can be achieved by use of either type of component alone.
  • the presence of mixtures of saturated and unsaturated fatty acids in the therapeutic wound healing composition significantly enhances the ability of pyruvate and the antioxidant to inhibit reactive oxygen production.
  • unsaturated fatty acids By stabilizing the cellular membrane, unsaturated fatty acids also improve membrane function and enhance pyruvate transport into the cell.
  • the three components in the therapeutic wound healing composition of the first aspect of Embodiment One (IA) function together in an unexpected synergistic manner to prevent and reduce injury to mammalian cells and increase the resuscitation rate of injured mammalian cells.
  • lactate is employed instead of an antioxidant. Antioxidants react with, and neutralize, oxygen radicals after the radicals are already formed. Lactate, on the other hand, is a component in the cellular feedback mechanism and inhibits the respiratory bursting process to suppress the production of active oxygen species.
  • the combination of pyruvate to neutralize active oxygen species and lactate to suppress the respiratory bursting process functions in a synergistic manner to reduce hydrogen peroxide production in the cell to levels lower than can be achieved by use of either type of component alone.
  • the presence of mixtures of saturated and unsaturated fatty acids in the therapeutic wound healing composition significantly enhances the ability of pyruvate and lactate to inhibit reactive oxygen production.
  • the three components in the therapeutic wound healing composition in the second aspect of Embodiment One (I.B) function together in a synergistic manner to protect and resuscitate mammalian cells.
  • the presence of mixtures of saturated and unsaturated fatty acids in the therapeutic wound healing composition in this embodiment significantly enhances the ability of the antioxidant to inhibit reactive oxygen production.
  • the combination of an antioxidant to neutralize active oxygen species and fatty acids to rebuild cellular membranes and reduce the need of the cell for oxygen functions in a synergistic manner to reduce hydrogen peroxide production in the cell to levels lower than can be achieved by either type of component alone.
  • the components in the therapeutic wound healing composition in the third aspect of Embodiment One (LC) function together in a synergistic manner to protect and resuscitate mammalian cells.
  • lactate is employed because the respiratory bursting process is more pronounced in monocytes than in epidermal keratinocytes.
  • lactate to suppress the respiratory bursting process and an antioxidant to neutralize active oxygen species functions in a synergistic manner to reduce hydrogen peroxide production in the cell to levels lower than can be achieved by either component alone.
  • the presence of mixtures of saturated and unsaturated fatty acids in the therapeutic wound healing composition in this embodiment significantly enhances the ability of lactate and the antioxidant to inhibit reactive oxygen production.
  • the three components in the therapeutic wound healing composition in the fourth aspect of Embodiment One (I.D) function together in an unexpected synergistic manner to protect and resuscitate mammalian cells.
  • the combination of ingredients set out in the above embodiments functions together in an enhanced manner to prevent and reduce injury to mammalian cells and increase the resuscitation rate of injured mammalian cells.
  • the therapeutic effect of the combination of the components in each of the above embodiments is markedly greater than that expected by the mere addition of the individual therapeutic components.
  • applicant's therapeutic wound healing compositions for treating mammalian cells have the ability to decrease intracellular levels of hydrogen peroxide production, increase cellular resistance to cytotoxic agents, increase rates of cellular proliferation, and increase cellular viability.
  • a therapeutic wound healing composition is made by forming an admixture of the components of the composition.
  • a therapeutic wound healing composition is made by forming an admixture of (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • a therapeutic wound healing composition is made by forming an admixture of (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • LC a therapeutic wound healing composition
  • a therapeutic wound healing composition is made by forming an admixture of (a) an antioxidant and (b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • a therapeutic wound healing composition is made by forming an admixture of (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
  • the admixture may be formed in a solvent such as water, and a surfactant may be added if required. If necessary, the pH of the solvent is adjusted to a range from about 3.5 to about 8.0, and preferably from about 4.5 to about 7.5, and more preferably about 6.0 to about 7.4. The admixture is then sterile filtered. Other ingredients may also be incorporated into the therapeutic wound healing composition as dictated by the nature of the desired composition as well known by those having ordinary skill in the art. The ultimate therapeutic wound healing compositions are readily prepared using methods generally known in the pharmaceutical arts.
  • the invention is directed to a method for preparing a therapeutic wound healing composition (IA) for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of admixing the following ingredients:
  • pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • an antioxidant an antioxidant
  • the present invention extends to methods for employing the therapeutic wound healing compositions of Embodiment One (I) in vivo and in vitro.
  • a therapeutic wound healing composition is employed by contacting the therapeutic composition with mammalian cells.
  • the invention is directed to a method for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of (A) providing a therapeutic wound healing composition which comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells, and (B) contacting the therapeutic wound healing composition with the mammalian cells.
  • a therapeutic wound healing composition which comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured ma
  • the invention is directed to a method for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of (A) providing a therapeutic wound healing composition which comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells, and (B) contacting the therapeutic wound healing composition with the mammalian cells.
  • a therapeutic wound healing composition which comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable
  • the invention is directed to a method for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of (A) providing a therapeutic wound healing composition which comprises (a) an antioxidant, and (b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells, and (B) contacting the therapeutic wound healing composition with the mammalian cells.
  • a therapeutic wound healing composition which comprises (a) an antioxidant, and (b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells
  • the invention is directed to a method for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of (A) providing a therapeutic wound healing composition which comprises (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells, and (B) contacting die therapeutic wound healing composition with the mammalian cells.
  • a therapeutic wound healing composition which comprises (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells, and (B) contacting die therapeutic wound
  • the invention is directed to a method for healing a wound in a mammal which comprises the steps of:
  • A providing a therapeutic wound healing composition (LA) which comprises: (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • the types of wounds which may be healed using the wound healing compositions of Embodiment One (LA-D) of the present invention are those which result from an injury which causes epidermal damage such as incisions, wounds in which the skin is broken by a cutting instrument, and lacerations, wounds in which the skin is broken by a dull or blunt instrument.
  • the therapeutic compositions may also be used to treat various dermatological disorders such as hyperkeratosis, photo-aging, bums, donor site wounds from skin transplants, ulcers (cutaneous, decubitus, venous stasis, and diabetic), psoriasis, skin rashes, and sunburn photoreactive processes.
  • the topical therapeutic compositions may also be used orally in the form of a mouth wash or spray to protect and accelerate the healing of injured oral tissue such as mouth sores and bums.
  • the topical therapeutic compositions may further be used in ophthalmological preparations to treat wounds such as those which result from comeal ulcers, radialkeratotomy, comeal transplants, epikeratophakia and other surgically induced wounds in the eye.
  • the topical therapeutic compositions may in addition be used in anorectal creams and suppositories to treat such conditions as pruritus and, proctitis, anal fissures, and hemorrhoids.
  • die therapeutic compositions are used to treat wounds such as incisions and lacerations.
  • the wound healing compositions of Embodiment One (LA-D) of the present invention may be utilized in topical products, ingestible products, and tissue culture medium to protect mammalian cells and increase the resuscitation rate of injured mammalian cells.
  • the therapeutic wound healing compositions may be used in topical skin care products to protect and increase the resuscitation rate of skin tissue such as in the treatment of various dermatological disorders such as hyperkeratosis, photo-aging, and sunburn photoreactive processes. Inju ⁇ y to skin can occur for a variety of reasons. Injury often occurs to individuals who wash their hands often, to individuals who are exposed to stressful environmental conditions (overexposure to sun or chemicals), or to the elderly or individuals with an underlining disease.
  • die wound healing compositions of the present invention provides a source of antioxidants to the skin which would protect die skin from the harmful effects of UV light, chemicals, and severe drying.
  • the wound healing compositions can be used for the following indications: a) Moisturizing and protecting; b) Healing dry cracked skin; c) Treating irritated skin such as diaper rash; d) Healing severe dry skin due to other diseases (venous dermatitis); e) Treating psoriasis and other hyperproliferative diseases; f) Protecting skin from UV light damage (antioxidant skin replacement); g) Treating sebo ⁇ heic conditions; and h) Treating shaving wounds in an after shave lotion.
  • the topical therapeutic wound healing compositions may also be used orally in the form of a mouth wash or spray to protect and accelerate the healing of injured oral tissue such as mouth sores and bu s.
  • the topical therapeutic wound healing compositions may further be used in ophthalmological preparations such as eye care products to neutralize hydrogen peroxide used in the cleaning of contact lenses.
  • the topical therapeutic wound healing compositions may in addition be used in anorectal creams and suppositories to treat such conditions as pruritus and, proctitis, anal fissures, and hemo ⁇ hoids.
  • white blood cells Initially as white blood cells enter a wound site, the cells release oxygen radicals, depleting die antioxidants at die wound site, thus impairing the healing process.
  • Incorporating the wound healing compositions of die present invention into a wound healing formulation would facilitate healing by providing die site with usable antioxidants, and a source of fatty acids needed for membrane repair.
  • the wound healing compositions can be used for the following indications: a) Healing of cuts and scrapes; b) Bu s (heals bums with less scaring and scabbing); c) Decubitus ulcers; d) Bed sores, pressure ulcers; e) Fissures, Hemo ⁇ hoids; f) Use in combination with immunostimulators (simulated healing in healing deficient people); g) Post surgical wounds; h) Bandages; i) Diabetic ulcers; j) Venous ulceration; and k) Use in combination with wound cleansing agents.
  • the therapeutic wound healing compositions may also be used in ingestible products to protect and increase the resuscitation rate of erosions, stomach ulcers, and hemo ⁇ hages in the gastric mucosa.
  • Other ingestible therapeutic products include: stroke medications; autoimmune disease medications; arthritis medications; ulcer medications; cancer medications (cytotoxic agents); heart medication to improve regional ventricular function and restore normal heart rate and pressure functions; lung medication to repair injured tissue; liver medication to suppress lipogenesis of alcoholic origin and prevent hepatic steatosis; kidney medication to suppress urinary calculi (kidney stones); detoxification medication to antagonize heavy metal poisoning, cyanide poisoning, sodium sulfide poisoning, other types of poisoning.
  • the therapeutic wound healing compositions may be used in ingestible products to treat inflammatory diseases such as hepatitis, gastritis, colitis, esophagitis, arthritis, and pancreatitis.
  • the therapeutic wound healing compositions of the present invention may also be used in tissue culture media and organ transplant media to prevent and reduce injury to mammalian cells and increase the resuscitation rate of injured mammalian cells.
  • Tissue cultures and transplant organs encounter reactive oxygen species generated in the culture media by die injured cells.
  • Organs particularly susceptible to oxidative damage during transport and transplantation due to reperfusion injury following ischemia are corneas, livers, hearts, and kidneys.
  • the tiierapeutic wound healing compositions may be useful to abrogate reperfusion injury to such transplant organs.
  • die invention is directed to a method for preserving mammalian cells in a culture medium which comprises the steps of:
  • (IA) (a) pyruvate selected from die group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures tiiereof;
  • LB (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; and (c) a mixture of saturated and unsaturated fatty acids wherein die fatty acids are tiiose fatty acids required for die repair of cellular membranes and resuscitation of mammalian cells;
  • LC (a) an antioxidant;
  • lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof;
  • step (C) contacting the therapeutic wound healing composition from step (A) witii die mammalian cells in the culture medium from step (B).
  • die inventive tiierapeutic wound healing compositions of Embodiment One may be stored for future use or may be formulated in effective amounts with pharmaceutically acceptable carriers to prepare a wide variety of pharmaceutical compositions.
  • pharmaceutically acceptable carriers are pharmaceutical appliances, topical vehicles (non-oral and oral), and ingestible vehicles.
  • Non-oral topical compositions employ non-oral topical vehicles, such as creams, gels formulations, foams, ointments and sprays, salves, and films, which are intended to be applied to die skin or body cavity and are not intended to be taken by mouth.
  • Oral topical compositions employ oral vehicles, such as mouthwashes, rinses, oral sprays, suspensions, and dental gels, which are intended to be taken by mouth but are not intended to be ingested.
  • Ingestible compositions employ ingestible or partly ingestible vehicles such as confectionery bulking agents which include hard and soft confectionery such as lozenges, tablets, toffees, nougats, suspensions, chewy candies, and chewing gums.
  • die therapeutic wound healing composition is incorporated into a pharmaceutical appliance which may be in the form of sutures, staples, gauze, bandages, bum dressings, artificial skins, liposome or micell formulations, microcapsules, aqueous vehicles for soaking gauze dressings, and the like, and mixtures thereof.
  • a pharmaceutical appliance which may be in the form of sutures, staples, gauze, bandages, bum dressings, artificial skins, liposome or micell formulations, microcapsules, aqueous vehicles for soaking gauze dressings, and the like, and mixtures thereof.
  • a variety of traditional ingredients may optionally be included in die pharmaceutical composition in effective amounts such as buffers, preservatives, tonicity adjusting agents, antioxidants, polymers for adjusting viscosity or for use as extenders, and excipients, and die like.
  • Such traditional ingredients include acetate and borate buffers; thimerosal, sorbic acid, methyl and propyl paraben and chlorobutanol preservatives; sodium chloride and sugars to adjust die tonicity; and excipients such as mannitol, lactose and sucrose.
  • Other conventional pharmaceutical additives known to those having ordinary skill in the pharmaceutical arts may also be used in the pharmaceutical composition.
  • tiierapeutically effective amounts of the therapeutic wound healing compositions of die present invention may be employed in die pharmaceutical appliance. These amounts are readily determined by tiiose skilled in the art widiout the need for undue experimentation.
  • the exact amount of the therapeutic wound healing composition employed is subject to such factors as the type and concentration of die therapeutic wound healing composition and d e type of pharmaceutical appliance employed.
  • the amount of tiierape ⁇ tic wound healing composition may be varied in order to obtain the result desired in the final product and such variations are within the capabilities of tiiose skilled in the art without die need for undue experimentation.
  • die pharmaceutical composition will comprise die tiierapeutic wound healing composition in an amount from about 0.1% to about 5%, by weight of the pharmaceutical composition. In a more preferred embodiment, the pharmaceutical composition will comprise die tiierapeutic wound healing composition in an amount from about 0.1% to about 3%, by weight of the pharmaceutical composition. In a most preferred embodiment, the pharmaceutical composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 1%, by weight of the pharmaceutical composition.
  • a pharmaceutical composition is made by contacting a therapeutically effective amount of a tiierapeutic wound healing composition with a pharmaceutical appliance and the other ingredients of die final desired pharmaceutical composition.
  • the therapeutic wound healing composition may be in a solvent and may be absorbed onto a pharmaceutical appliance.
  • compositions will usually be incorporated into the composition as dictated by the nature of the desired composition as well known by those having ordinary skill in the art.
  • the ultimate pharmaceutical compositions are readily prepared using methods generally known in the pharmaceutical arts.
  • die therapeutic wound healing composition is incorporated into a non-oral topical vehicle which may be in the form of a cream, gel, foam, ointment, spray, and die like.
  • Typical non-toxic non-oral topical vehicles known in the pharmaceutical arts may be used in die present invention.
  • the prefe ⁇ ed non-oral topical vehicles are water and pharmaceutically acceptable water- miscible organic solvents such as ethyl alcohol, isopropyl alcohol, propylene glycol, glycerin, and the like, and mixtures of these solvents.
  • Water-alcohol mixtures are particularly preferred and are generally employed in a weight ratio from about 1 : 1 to about 20:1, preferably from about 3:1 to about 20:1, and most preferably from about 3:1 to about 10:1, respectively.
  • the non-oral topical therapeutic wound healing compositions may also contain conventional additives employed in tiiose products.
  • Conventional additives include humectants, emollients, lubricants, stabilizers, dyes, and perfumes, providing the additives do not interfere with the therapeutic properties of the tiierapeutic wound healing composition.
  • Suitable humectants useful in the non-oral topical therapeutic wound healing compositions include glycerin, propylene glycol, polyethylene glycol, sorbitan, fructose, and die like, and mixtures thereof. Humectants, when employed, may be present in amounts from about 10% to about 20%, by weight of the topical therapeutic wound healing composition.
  • the coloring agents (colors, colorants) useful in the non-oral topical therapeutic wound healing composition are used in amounts effective to produce die desired color.
  • These coloring agents include pigments which may be incorporated in amounts up to about 6% by weight of the non-oral topical therapeutic wound healing composition.
  • a prefe ⁇ ed pigment, titanium dioxide may be incorporated in amounts up to about 2%, and preferably less than about 1 %, by weight of the non-oral topical therapeutic wound healing composition.
  • the coloring agents may also include natural food colors and dyes suitable for food, drug and cosmetic applications. These coloring agents are known as F.D.& C. dyes and lakes.
  • the materials acceptable for the foregoing uses are preferably water-soluble.
  • Illustrative nonlimiting examples include die indigoid dye known as F.D.& C.
  • F.D.& C. Green No.l comprises a triphenylmethane dye and is die monosodium salt of 4-[4-(N-ethyl- ⁇ >- sulfoniumbenzylamino) diphenylmethylene]-[ 1 -(N-ethyl-N-g-sulfoniumbenzyl)-delta- 2,5-cyclohexadieneimine].
  • tiierapeutically effective amounts of the therapeutic wound healing compositions of die present invention may be admixed witii a non-oral topical vehicle to form a topical therapeutic wound healing composition.
  • die non-oral topical therapeutic wound healing compositions will comprise die therapeutic wound healing composition in an amount from about 0.1% to about 10% and a non-oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the non-oral topical therapeutic wound healing composition.
  • the non-oral topical therapeutic wound healing compositions will comprise die therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most preferred embodiment, the non-oral topical therapeutic wound healing compositions will comprise the tiierapeutic wound healing composition in an amount from about 0.1% to about 2%, and a non-oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the non-oral topical tiierapeutic wound healing composition.
  • the present invention extends to metiiods for preparing the non-oral topical therapeutic wound healing compositions.
  • die non-oral topical tiierapeutic wound healing composition is prepared by admixing a tiierapeutically effective amount of the therapeutic wound healing composition of die present invention and a non-oral topical vehicle.
  • the final compositions are readily prepared using standard methods and apparatus generally known by those skilled in die pharmaceutical arts.
  • the apparatus useful in accordance witii the present invention comprises mixing apparatus well known in the pharmaceutical arts, and therefore the selection of the specific apparatus will be apparent to the artisan.
  • the therapeutic wound healing composition is incorporated into an oral topical vehicle which may be in the form of a mouthwash, rinse, oral spray, suspension, dental gel, and die like.
  • an oral topical vehicle which may be in the form of a mouthwash, rinse, oral spray, suspension, dental gel, and die like.
  • Typical non-toxic oral vehicles known in the pharmaceutical arts may be used in die present invention.
  • the preferred oral vehicles are water, etiianol, and water-etiianol mixtures.
  • the water- ethanol mixtures are generally employed in a weight ratio from about 1:1 to about 20:1, preferably from about 3:1 to about 20:1, and most preferably from about 3:1 to about 10:1, respectively.
  • the pH value of the oral vehicle is generally from about 4 to about 7, and preferably from about 5 to about 6.5.
  • An oral topical vehicle having a pH value below about 4 is generally irritating to the oral cavity and an oral vehicle having a pH value greater than about 7 generally results in an unpleasant mouth feel.
  • the oral topical therapeutic wound healing compositions may also contain conventional additives normally employed in tiiose products.
  • Conventional additives include a fluorine providing compound, a sweetening agent, a flavoring agent, a coloring agent, a humectant, a buffer, and an emulsifier, providing the additives do not interfere with the therapeutic properties of the therapeutic wound healing composition.
  • coloring agents and humectants, and die amounts of these additives to be employed, set out above as useful in the non-oral topical therapeutic wound healing composition may be used in the oral topical therapeutic wound healing composition.
  • Fluorine providing compounds may be fully or slightly water soluble and are characterized by their ability to release fluoride ions or fluoride containing ions in water and by tiieir lack of reaction with other components in the composition.
  • Typical fluorine providing compounds are inorganic fluoride salts such as water-soluble alkali metal, alkaline earth metal, and heavy metal salts, for example, sodium fluoride, potassium fluoride, ammonium fluoride, cuprous fluoride, zinc fluoride, stannic fluoride, stannous fluoride, barium fluoride, sodium fluorosilicate, ammonium fluorosilicate, sodium fluorozirconate, sodium monofluorophosphate, aluminum mono- and di-fluorophosphates and fluorinated sodium calcium pyrophosphate.
  • inorganic fluoride salts such as water-soluble alkali metal, alkaline earth metal, and heavy metal salts, for example, sodium fluoride, potassium fluoride, ammonium flu
  • Alkali metal fluorides, tin fluoride and monofluorophosphates, such as sodium and stannous fluoride, sodium monofluorophosphate and mixtures thereof, are prefe ⁇ ed.
  • the amount of fluorine providing compound present in d e present oral topical therapeutic wound healing composition is dependent upon the type of fluorine providing compound employed, die solubility of the fluorine compound, and die nature of the final oral therapeutic wound healing composition.
  • the amount of fluorine providing compound used must be a nontoxic amount. In general, the fluorine providing compound when used will be present in an amount up to about 1%, preferably from about 0.001% to about 0.1%, and most preferably from about 0.001% to about 0.05%, by weight of the oral topical therapeutic wound healing composition.
  • sweetening agents When sweetening agents (sweeteners) are used, tiiose sweeteners well known in the art, including both natural and artificial sweeteners, may be employed.
  • the sweetening agent used may be selected from a wide range of materials including water-soluble sweetening agents, water-soluble artificial sweetening agents, water- soluble sweetening agents derived from naturally occurring water-soluble sweetening agents, dipeptide based sweetening agents, and protein based sweetening agents, including mixtures thereof.
  • representative categories and examples include:
  • water-soluble sweetening agents such as monosaccharides, disaccharides and polysaccharides such as xylose, ribose, glucose (dextrose), mannose, galactose, fructose (levulose), sucrose (sugar), maltose, invert sugar (a mixture of fructose and glucose derived from sucrose), partially hydrolyzed starch, co syrup solids, dihydrochalcones, monellin, steviosides, and glycy ⁇ hizin, and mixtures thereof;
  • water-soluble sweetening agents such as monosaccharides, disaccharides and polysaccharides such as xylose, ribose, glucose (dextrose), mannose, galactose, fructose (levulose), sucrose (sugar), maltose, invert sugar (a mixture of fructose and glucose derived from sucrose), partially hydrolyzed starch, co syrup solids, dihydrochalcon
  • water-soluble artificial sweeteners such as soluble saccharin salts, i.e., sodium or calcium saccharin salts, cyclamate salts, the sodium, ammonium or calcium salt of 3 ,4-dihydro-6-methyl- 1 ,2,3-oxathiazine-4-one-2,2-dioxide, die potassium salt of 3 ,4-dihydro-6-med ⁇ yl- 1 ,2 ,3-oxathiazine-4-one-2,2-dioxide ( Acesulfame-K), the free acid form of saccharin, and the like;
  • soluble saccharin salts i.e., sodium or calcium saccharin salts, cyclamate salts, the sodium, ammonium or calcium salt of 3 ,4-dihydro-6-methyl- 1 ,2,3-oxathiazine-4-one-2,2-dioxide, die potassium salt of 3 ,4-dihydro-6-med ⁇ yl- 1
  • dipeptide based sweeteners such as L-aspartic acid derived sweeteners, such as L-aspartyl-L-phenylalanine methyl ester (Aspartame) and materials described in United States Patent No. 3,492,131 , L-Alpha-aspa ⁇ tyl-N-(2,2,4,4- tetramethyl-3-thietanyl)-D-alanin-amide hydrate (Alitame), methyl esters of L-aspartyl- L-phenylglycerineandL-aspartyl-L-2,5-dihydrophenyl-glycine, L-aspartyl-2,5-dihydro- L-phenylalanine; L-aspartyl-L-(l-cyclohexen)-alanine, and die like;
  • L-aspartic acid derived sweeteners such as L-aspartyl-L-phenylalanine methyl ester (Aspartame) and materials described
  • water-soluble sweeteners derived from naturally occurring water- soluble sweeteners such as chlorinated derivatives of ordinary sugar (sucrose), e.g., chlorodeoxysugar derivatives such as derivatives of chlorodeoxysucrose or chlorodeoxygalactosucrose, known, for example, under the product designation of Sucralose;
  • chlorodeoxysucrose and chlorodeoxygalacto-sucrose derivatives include but are not limited to: l-chloro-l'-deoxysucrose; 4-chloro-4-deoxy-Alpha-D- galacto-pyranosyl-Alpha-D-fructofuranoside, or 4-chloro-4-deoxygalactosucrose; 4- chloro-4-deoxy-Alpha-D-galacto-pyranosyl- 1 -chloro- 1 -deoxy-B-D-fructo-furanoside, or
  • an effective amount of sweetening agent is utilized to provide the level of sweetness desired in die particular oral topical therapeutic wound healing composition, and this amount will vary with die sweetener selected and die final oral therapeutic product desired.
  • the amount of sweetener normally present is in the range from about 0.0025% to about 90%, by weight of the oral topical tiierapeutic wound healing composition, depending upon the sweetener used.
  • the exact range of amounts for each type of sweetener is well known in the art and is not the subject of the present invention.
  • flavoring agents include tiiose flavors known to the skilled artisan, such as natural and artificial flavors.
  • Suitable flavoring agents include mints, such as peppermint, cirrus flavors such as orange and lemon, artificial vanilla, cinnamon, various fruit flavors, both individual and mixed, and die like.
  • the amount of flavoring agent employed in the oral topical therapeutic wound healing composition is normally a matter of preference subject to such factors as the type of final oral therapeutic wound healing composition, the individual flavor employed, and die strength of flavor desired. Thus, the amount of flavoring may be varied in order to obtain the result desired in the final product and such variations are within the capabilities of those skilled in the art without the need for undue experimentation.
  • the flavoring agents, when used, are generally utilized in amounts tiiat may, for example, range in amounts from about 0.05% to about 6%, by weight of the oral topical therapeutic wound healing composition.
  • Suitable buffer solutions useful in die non-oral topical tiierapeutic wound healing compositions include citric acid-sodium citrate solution, phosphoric acid- sodium phosphate solution, and acetic acid-sodium acetate solution in amounts up to about 1%, and preferably from about 0.05% to about 0.5% by weight of the oral topical therapeutic wound healing composition.
  • therapeutically effective amounts of the therapeutic wound healing compositions of the present invention may be admixed witii an oral topical vehicle to form a topical therapeutic wound healing composition.
  • these amounts are readily determined by tiiose skilled in the art without the need for undue experimentation.
  • die oral topical therapeutic wound healing compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 10% and a oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the oral topical therapeutic wound healing composition.
  • the oral topical therapeutic wound healing compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most preferred embodiment, the oral topical therapeutic wound healing compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and a oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the oral topical tiierapeutic wound healing composition.
  • the present invention extends to methods for preparing the oral topical therapeutic wound healing compositions.
  • die oral topical therapeutic wound healing composition is prepared by admixing a therapeutically effective amount of the therapeutic wound healing composition of the present invention and an oral topical vehicle.
  • the final compositions are readily prepared using standard methods and apparatus generally known by those skilled in the pharmaceutical arts.
  • the apparatus useful in accordance with the present invention comprises mixing apparatus well known in the pharmaceutical arts, and tiierefore the selection of the specific apparatus will be apparent to the artisan.
  • an oral topical therapeutic wound healing composition is made by first dissolving coloring agents, sweetening agents, and similar additives in water.
  • the therapeutic wound healing composition is then admixed witii the aqueous solution. Then sufficient water or ethanol, or mixtures of water and ethanol, are added to the solution with mixing until the final solution volume is reached.
  • the therapeutic wound healing composition is added to die solution as the final ingredient.
  • the final oral topical therapeutic wound healing compositions are readily prepared using metiiods generally known in the pharmaceutical arts.
  • the oral therapeutic wound healing composition may also be in the form of dental gel.
  • gel means a solid or semisolid colloid which contains considerable quantities of water.
  • the colloid particles in a gel are linked together in a coherent meshwork which immobilizes the water contained inside d e meshwork.
  • the dental gel compositions of the present invention may contain the conventional additives set out above for oral topical therapeutic wound healing compositions such as mouthwashes, rinses, oral sprays, and suspensions and, in addition, may contain additional additives such as a polishing agent, a desensitizing agent, and die like, providing die additional additives do not interfere with die tiierapeutic properties of the therapeutic wound healing composition.
  • the oral vehicle In a dental gel composition, the oral vehicle generally comprises water, typically in an amount from about 10% to about 90%, by weight of the dental gel composition.
  • Polyethylene glycol, propylene glycol, glycerin, and mixtures thereof may also be present in the vehicle as humectants or binders in amounts from about 18% to about 30%, by weight of the dental gel composition.
  • Particularly preferred oral vehicles comprise mixtures of water with polyethylene glycol or water with glycerin and polypropylene glycol.
  • the dental gels of die present invention include a gelling agent (diickening agent) such as a natural or synthetic gum or gelatin.
  • Gelling agents such as hydroxyethyl cellulose, methyl cellulose, glycerin, carboxypolymethylene, and gelatin and die like, and mixtures thereof may be used.
  • the preferred gelling agent is hydroxyediyl cellulose. Gelling agents may be used in amounts from about 0.5% to about 5%, and preferably from about 0.5% to about 2%, by weight of the dental gel composition.
  • the dental gel compositions of the present invention may also include a polishing agent.
  • a polishing agent of colloidal silica and/or alkali metal aluminosilicate complexes is prefe ⁇ ed since these materials have refractive indices close to the refractive indices of the gelling systems commonly used in dental gels.
  • a polishing agent of calcium carbonate or calcium dihydrate may be used. These polishing agents may be used in amounts up to about 75%, and preferably in amounts up to about 50%, by weight of the dental gel composition.
  • the dental gel may also contain a desensitizing agent such as a combination of citric acid and sodium citrate.
  • Citric acid may be used in an amount from about 0.1% to about 3%, and preferably from about 0.2% to about 1%, by weight, and sodium citrate may be used in an amount from about 0.3% to about 9%, and preferably from about 0.6% to about 3%, by weight of the dental gel composition.
  • therapeutically effective amounts of the therapeutic wound healing compositions of the present invention may be admixed into the dental gel compositions. These amounts are readily determined by tiiose skilled in the art without the need for undue experimentation.
  • die dental gel compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 10% and an oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the dental gel composition.
  • die dental gel compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most prefe ⁇ ed embodiment, the dental gel compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and an oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the dental gel composition.
  • the present invention extends to methods for preparing the therapeutic dental gel compositions.
  • the dental gel composition is prepared by admixing a tiierapeutically effective amount of the therapeutic wound healing composition of die present invention and an oral topical vehicle.
  • the final compositions are readily prepared using methods generally known by those skilled in the dental and pharmaceutical arts.
  • the apparatus useful in accordance witii the present invention comprises mixing apparatus well known in the pharmaceutical arts, and therefore the selection of the specific apparatus will be apparent to the artisan.
  • a therapeutic dental gel composition is made by first dispersing a gelling agent in a humectant or water, or a mixture of both, then admixing to the dispersion an aqueous solution of the water-soluble additives such as the fluorine providing compound, sweeteners and die like, then adding die polishing agent, and lastly admixing the flavoring agent and the therapeutic wound healing composition.
  • the final gel mixture is then tubed or otherwise packaged.
  • the liquids and solids in a gel product are proportioned to form a creamy or gelled mass which is extrudable from a pressurized container or from a collapsible tube.
  • the final therapeutic wound healing compositions are readily prepared using metiiods generally known in the pharmaceutical arts.
  • the therapeutic wound healing composition is incorporated into an ingestible vehicle.
  • the ingestible vehicle may be a confectionery bulking agent in the form of lozenges, tablets, toffees, nougats, suspensions, chewy candies, chewing gums, and die like.
  • the pharmaceutically acceptable carriers may be prepared from a wide range of materials including, but not limited to, diluents, binders and adhesives, lubricants, disintegrants, coloring agents, bulking agents, flavoring agents, sweetening agents and miscellaneous materials such as buffers and adsorbents tiiat may be needed in order to prepare a particular therapeutic confection.
  • confectionery formulations are historically well known and has changed little through the years. Confectionery items have been classified as eitiier "hard” confectionery or "soft” confectionery.
  • the therapeutic wound healing compositions of the present invention can be incorporated into confectionery compositions by admixing the inventive composition into conventional hard and soft confections.
  • die term confectionery material means a product containing a bulking agent selected from a wide variety of materials such as sugar, com syrup, and in the case of sugarless bulking agents, sugar alcohols such as sorbitol and mannitol and mixtures thereof.
  • Confectionery material may include such exemplary substances as lozenges, tablets, toffee, nougat, suspensions, chewy candy, chewing gum and the like.
  • the bulking agent is present in a quantity sufficient to bring the total amount of composition to 100%. In general, the bulking agent will be present in amounts up to about 99.98%, preferably in amounts up to about 99.9%, and more preferably in amounts up to about 99%, by weight of the ingestible therapeutic wound healing composition.
  • Lozenges are flavored medicated dosage forms intended to be sucked and held in the mouth. Lozenges may be in the form of various shapes such as flat, circular, octagonal and biconvex forms.
  • the lozenge bases are generally in two forms: hard boiled candy lozenges and compressed tablet lozenges.
  • Hard boiled candy lozenges may be processed and formulated by conventional means.
  • a hard boiled candy lozenge has a base composed of a mixture of sugar and other carbohydrate bulking agents kept in an amorphous or glassy condition.
  • This amorphous or glassy form is considered a solid syrup of sugars generally having from about 0.5% to about 1.5% moisture.
  • Such materials normally contain up to about 92% com syrup, up to about 55% sugar and from about 0.1% to about 5% water, by weight of the final composition.
  • the syrup component is generally prepared from com syrups high in fructose, but may include other materials. Further ingredients such as flavoring agents, sweetening agents, acidulants, coloring agents and die like may also be added.
  • Boiled candy lozenges may also be prepared from non-fermentable sugars such as sorbitol, mannitol, and hydrogenated com syrup.
  • Typical hydrogenated co syrups are Lycasin, a commercially available product manufactured by Roquette Corporation, and Hystar, a commercially available product manufactured by Lonza, Inc.
  • the candy lozenges may contain up to about 95% sorbitol, a mixture of sorbitol and mannitol in a ratio from about 9.5:0.5 up to about 7.5:2.5, and hydrogenated com syrup up to about 55%, by weight of the solid syrup component.
  • Boiled candy lozenges may be routinely prepared by conventional methods such as those involving fire cookers, vacuum cookers, and scraped-surface cookers also refe ⁇ ed to as high speed atmospheric cookers.
  • Fire cookers involve the traditional method of making a boiled candy lozenge base. In this method, the desired quantity of carbohydrate bulking agent is dissolved in water by heating d e agent in a kettle until the bulking agent dissolves. Additional bulking agent may then be added and cooking continued until a final temperature of 145°C. to 156°C. is achieved. The batch is then cooled and worked as a plastic-like mass to incorporate additives such as flavors, colorants and die like.
  • a high-speed atmospheric cooker uses a heat-exchanger surface which involves spreading a film of candy on a heat exchange surface, the candy is heated to
  • the candy is tiien rapidly cooled to 100°C. to 120°C. and worked as a plastic-like mass enabling incorporation of the additives, such as flavors, colorants and die like.
  • the carbohydrate bulking agent is boiled to 125°C. to 132°C, vacuum is applied and additional water is boiled off without extra heating.
  • the mass is a semi-solid and has a plastic-like consistency.
  • flavors, colorants, and other additives are admixed in the mass by routine mechanical mixing operations.
  • the optimum mixing required to uniformly mix the flavoring agents, coloring agents and other additives during conventional manufacturing of boiled candy lozenges is determined by the time needed to obtain a uniform distribution of the materials. Normally, mixing times of from 4 to 10 minutes have been found to be acceptable.
  • the boiled candy lozenge may be cut into workable portions or formed into desired shapes.
  • a variety of forming techniques may be utilized depending upon the shape and size of the final product desired.
  • a general discussion of the composition and preparation of hard confections may be found in HA. Lieberman, Pharmaceutical Dosage Forms: Tablets, Volume I (1 80), Marcel Dekker, Inc., New York, N.Y. at pages 339 to 469, which disclosure is incorporated herein by reference.
  • the apparatus useful in accordance witii the present invention comprises cooking and mixing apparatus well known in the confectionery manufacturing arts, and dierefore the selection of the specific apparatus will be apparent to the artisan.
  • compressed tablet confections contain particulate materials and are formed into structures under pressure.
  • These confections generally contain sugars in amounts up to about 95%, by weight of the composition, and typical tablet excipients such as binders and lubricants as well as flavoring agents, coloring agents and the like.
  • the lozenges of the present invention may be made of soft confectionery materials such as those contained in nougat.
  • the preparation of soft confections, such as nougat involves conventional methods, such as the combination of two primary components, namely (1) a high boiling syrup such as a co syrup, hydrogenated starch hydrolysate or the like, and (2) a relatively light textured frappe, generally prepared from egg albumin, gelatin, vegetable proteins, such as soy derived compounds, sugarless milk derived compounds such as milk proteins, and mixtures thereof.
  • the frappe is generally relatively light, and may, for example, range in density from about 0.5 to about 0.7 grams/cc.
  • the high boiling syrup, or "bob syrup” of the soft confectionery is relatively viscous and has a higher density than the frappe component, and frequently contains a substantial amount of carbohydrate bulking agent such as a hydrogenated starch hydrolysate.
  • carbohydrate bulking agent such as a hydrogenated starch hydrolysate.
  • the final nougat composition is prepared by the addition of the "bob syrup” to the frappe under agitation, to form the basic nougat mixture. Further ingredients such as flavoring agents, additional carbohydrate bulking agent, coloring agents, preservatives, medicaments, mixtures thereof and the like may be added thereafter also under agitation.
  • Further ingredients such as flavoring agents, additional carbohydrate bulking agent, coloring agents, preservatives, medicaments, mixtures thereof and the like may be added thereafter also under agitation.
  • a general discussion of the composition and preparation of nougat confections may be found in B.W. Minifie, Chocolate, Cocoa and Confectionery: Science and Technology. 2nd edition, AVI Publishing Co., Inc.
  • the procedure for preparing the soft confectionery involves known procedures.
  • the frappe component is prepared first and thereafter the syrup component is slowly added under agitation at a temperature of at least about 65°C, and preferably at least about 100°C.
  • the mixture of components is continued to be mixed to form a uniform mixture, after which the mixture is cooled to a temperature below
  • the flavoring agent may be added.
  • the mixture is further mixed for an additional period until it is ready to be removed and formed into suitable confectionery shapes.
  • the ingestible therapeutic wound healing compositions may also be in the form of a pharmaceutical suspension.
  • Pharmaceutical suspensions of this invention may be prepared by conventional methods long established in die art of pharmaceutical compounding. Suspensions may contain adjunct materials employed in formulating the suspensions of the art.
  • the suspensions of the present invention can comprise: (a) preservatives such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), benzoic acid, ascorbic acid, metiiyl paraben, propyl paraben, tocopherols, and die like, and mixtures thereof.
  • Preservatives are generally present in amounts up to about 1%, and preferably from about 0.05% to about 0.5%, by weight of the suspension; (b) buffers such as citric acid-sodium citrate, phosphoric acid-sodium phosphate, and acetic acid-sodium acetate in amounts up to about 1%, and preferably from about 0.05% to about 0.5%, by weight of the suspension;
  • suspending agents or thickeners such as cellulosics like methylcellulose, carrageenans like alginic acid and its derivatives, xanthan gums, gelatin, acacias, and microciystalline cellulose in amounts up to about 20%, and preferably from about 1% to about 15%, by weight of the suspension;
  • antifoaming agents such as dimethyl polysiloxane in amounts up to about 0.2%, and preferably from about 0.01% to about 0.1%, by weight of the suspension;
  • sweetening agents such as those sweeteners well known in the art, including both natural and artificial sweeteners.
  • Sweetening agents such as monosaccharides, disaccharides and polysaccharides such as xylose, ribose, glucose (dextrose), mannose, galactose, fructose (levulose), sucrose (sugar), maltose, invert sugar (a mixture of fructose and glucose derived from sucrose), partially hydrolyzed starch, com syrup solids, dihydrochalcones, monellin, steviosides, glycyrrhizin, and sugar alcohols such as sorbitol, mannitol, maltitol, hydrogenated starch hydrolysates and mixtures thereof may be utilized in amounts up to about 60%, and preferably from about 20% to about 50%, by weight of the suspension.
  • Water-soluble artificial sweeteners such as soluble saccharin salts, i.e., sodium or calcium saccharin salts, cyclamate salts, the sodium, ammonium or calcium salt of 3,4-dihydro-6-methyl-l,2,3- oxathiazine-4-one-2,2-dioxide, the potassium salt of 3,4-dihydro-6-methyl- 1,2,3 - oxathiazine-4-one-2,2-dioxide (Acesulfame-K), the free acid form of saccharin, and the like may be utilized in amounts from about 0.001% to about 5%, by weight of the suspension;
  • flavoring agents such as those flavors well known to the skilled artisan, such as natural and artificial flavors and mints, such as peppermint, menthol, citrus flavors such as orange and lemon, artificial vanilla, cinnamon, various fruit flavors, both individual and mixed and die like may be utilized in amounts from about 0.5% to about 5%, by weight of the suspension;
  • coloring agents such as pigments which may be incorporated in amounts up to about 6%, by weight of the suspension.
  • a prefe ⁇ ed pigment, titanium dioxide may be incorporated in amounts up to about 2%, and preferably less than about 1%, by weight of the suspension.
  • the coloring agents may also include natural food colors and dyes suitable for food, drug and cosmetic applications. These colorants are known as F.D.& C. dyes and lakes.
  • the materials acceptable for the foregoing uses are preferably water-soluble.
  • Such dyes are generally present in amounts up to about 0.25%, and preferably from about 0.05% to about 0.2%, by weight of the suspension;
  • decolorizing agents such as sodium metabisulfite, ascorbic acid and the like may be incorporated into the suspension to prevent color changes due to aging.
  • decolorizing agents may be used in amounts up to about 0.25%, and preferably from about 0.05% to about 0.2%, by weight of the suspension;
  • solubilizers such as alcohol, propylene glycol, polyethylene glycol, and die like may be used to solubilize the flavoring agents.
  • solubilizing agents may be used in amounts up to about 10%, and preferably from about 2% to about 5%, by weight of the suspension.
  • the pharmaceutical suspensions of the present invention may be prepared as follows:
  • (A) admix the thickener with water heated from about 40°C. to about 95°C, preferably from about 40°C. to about 70°C, to form a dispersion if the thickener is not water soluble or a solution if the thickener is water soluble;
  • step (D) combine the sweetener solution with the thickener-therapeutic wound healing composition and mix until uniform; and (E) admix the optional adjunct materials such as coloring agents, flavoring agents, decolorants, solubilizers, antifoaming agents, buffers and additional water with the mixture of step (D) to form the suspension.
  • optional adjunct materials such as coloring agents, flavoring agents, decolorants, solubilizers, antifoaming agents, buffers and additional water
  • the ingestible therapeutic wound healing compositions of this invention may also be in chewable form.
  • these considerations include die amount of active substance per tablet, the flavoring agent employed, the degree of compressibility of the tablet and die organoleptic properties of the composition.
  • Chewable therapeutic candy is prepared by procedures similar to those used to make soft confectionery.
  • a boiled sugar-corn syrup blend is formed to which is added a frappe mixture.
  • the boiled sugar-corn syrup blend may be prepared from sugar and com syrup blended in parts by weight ratio of about 90:10 to about 10:90.
  • the sugar-corn syrup blend is heated to temperatures above about 120°C. to remove water and to form a molten mass.
  • the frappe is generally prepared from gelatin, egg albumin, milk proteins such as casein, and vegetable proteins such as soy protein, and the like, which is added to a gelatin solution and rapidly mixed at ambient temperature to form an aerated sponge like mass.
  • the frappe is then added to the molten candy mass and mixed until homogeneous at temperatures between about 65°C. and about 120°C.
  • the ingestible therapeutic wound healing composition of the instant invention can then be added to the homogeneous mixture as the temperature is lowered to about 65°C.-95°C. whereupon additional ingredients can then be added such as flavoring agents and coloring agents.
  • additional ingredients can then be added such as flavoring agents and coloring agents.
  • the formulation is further cooled and formed into pieces of desired dimensions.
  • therapeutically effective amounts of the therapeutic wound healing compositions of the present invention may be admixed into the hard and soft confectionery products. These amounts are readily determined by those skilled in the art without the need for undue experimentation.
  • die ingestible therapeutic wound healing composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 10% and an ingestible vehicle, that is a pharmaceutically acceptable carrier, in a quantity sufficient to bring the total amount of composition to 100%, by weight the ingestible therapeutic wound healing composition.
  • the ingestible composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most prefe ⁇ ed embodiment, the ingestible composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and an ingestible vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight the ingestible therapeutic wound healing composition.
  • the present invention extends to metiiods of making the ingestible therapeutic wound healing compositions.
  • an ingestible therapeutic wound healing composition is prepared by admixing a therapeutically effective amount of the therapeutic wound healing composition with a pharmaceutically-acceptable carrier.
  • the apparatus useful in accordance with the present invention comprises mixing and heating apparatus well known in the confectionery arts, and therefore the selection of the specific apparatus will be apparent to the artisan.
  • the final ingestible therapeutic wound healing compositions are readily prepared using methods generally known in the confectionery arts.
  • the therapeutic wound healing compositions may also be incorporated into chewing gums.
  • the chewing gum composition contains a gum base, a bulking agent, the inventive therapeutic wound healing composition, and various additives.
  • the gum base employed will vary greatly depending upon various factors such as the type of base desired, die consistency of gum desired and die odier components used in die composition to make the final chewing gum product.
  • the gum base may be any water-insoluble gum base known in the art, and includes tiiose gum bases utilized for chewing gums and bubble gums.
  • suitable polymers in gum bases include botii natural and synthetic elastomers and rubbers.
  • those polymers which are suitable as gum bases include, without limitation, substances of vegetable origin such as chicle, crown gum, nispero, rosadinha, jelutong, perillo, niger gutta, tunu, balata, gutta-percha, lechi-capsi, sorva, gutta kay, mixtures thereof and die like.
  • Synthetic elastomers such as butadiene-styrene copolymers, polyisobutylene, isobutylene-isoprene copolymers, polyethylene, mixtures thereof and the like are particularly useful.
  • the gum base may include a non-toxic vinyl polymer, such as polyvinyl acetate and its partial hydrolysate, polyvinyl alcohol, and mixtures thereof. When utilized, die molecular weight of the vinyl polymer may range from about 2,000 up to and including about 94,000.
  • the amount of gum base employed will vary greatly depending upon various factors such as the type of base used, die consistency of the gum desired and the other components used in the composition to make the final chewing gum product. In general, the gum base will be present in amounts from about 5% to about 94%, by weight of the final chewing gum composition, and preferably in amounts from about
  • the gum base composition may contain conventional elastomer solvents to aid in softening the elastomer base component.
  • elastomer solvents may comprise terpinene resins such as polymers of Alpha-pinene or ⁇ -pinene, methyl, glycerol or pentaerythritol esters of rosins or modified rosins and gums, such as hydrogenated, dimerized or polymerized rosins or mixtures thereof.
  • elastomer solvents suitable for use herein include die pentaerythritol ester of partially hydrogenated wood or gum rosin, the pentaerythritol ester of wood or gum rosin, the glycerol ester of wood rosin, the glycerol ester of partially dimerized wood or gum rosin, the glycerol ester of polymerized wood or gum rosin, the glycerol ester of tall oil rosin, the glycerol ester of wood or gum rosin and the partially hydrogenated wood or gum rosin and die partially hydrogenated methyl ester of wood or rosin, mixtures thereof, and the like.
  • the elastomer solvent may be employed in amounts from about 5% to about 75%, by weight of the gum base, and preferably from about 45% to about 70%, by weight of the gum base.
  • a variety of traditional ingredients may be included in the gum base in effective amounts such as plasticizers or softeners such as lanolin, palmitic acid, oleic acid, stearic acid, sodium stearate, potassium stearate, glyceryl triacetate, glyceryl lecithin, glyceryl monostearate, propylene glycol monostearate, acetylated monoglyceride, glycerine, mixtures thereof, and the like may also be incorporated into die gum base to obtain a variety of desirable textures and consistency properties.
  • plasticizers or softeners such as lanolin, palmitic acid, oleic acid, stearic acid, sodium stearate, potassium stearate, glyceryl triacetate, glyceryl lecithin, glyceryl monostearate, propylene glycol monostearate, acetylated monoglyceride, glycerine, mixtures thereof, and the like may also
  • Waxes for example, natural and synthetic waxes, hydrogenated vegetable oils, petroleum waxes such as polyurethane waxes, polyethylene waxes, paraffin waxes, microcrystalline waxes, fatty waxes, sorbitan monostearate, tallow, propylene glycol, mixtures thereof, and the like may also be incorporated into the gum base to obtain a variety of desirable textures and consistency properties.
  • These traditional additional materials are generally employed in amounts up to about 30%, by weight of the gum base, and preferably in amounts from about 3% to about 20%, by weight of die gum base.
  • the gum base may include effective amounts of mineral adjuvants such as calcium carbonate, magnesium carbonate, alumina, aluminum hydroxide, aluminum silicate, talc, tricalcium phosphate, dicalcium phosphate and die like as well as mixtures thereof. These mineral adjuvants may serve as fillers and textural agents. These fillers or adjuvants may be used in the gum base in various amounts. Preferably the amount of filler when used will be present in an amount up to about 60%, by weight of the chewing gum base.
  • mineral adjuvants such as calcium carbonate, magnesium carbonate, alumina, aluminum hydroxide, aluminum silicate, talc, tricalcium phosphate, dicalcium phosphate and die like as well as mixtures thereof. These mineral adjuvants may serve as fillers and textural agents. These fillers or adjuvants may be used in the gum base in various amounts. Preferably the amount of filler when used will be present in an amount up to about 60%, by weight of the chewing gum base.
  • the chewing gum base may additionally include the conventional additives of coloring agents, antioxidants, preservatives and die like.
  • coloring agents for example, titanium dioxide and other dyes suitable for food, drug and cosmetic applications, known as F.D. & C. dyes, may be utilized.
  • An antioxidant such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, and mixtures thereof, may also be included.
  • BHT butylated hydroxytoluene
  • BHA butylated hydroxyanisole
  • propyl gallate propyl gallate
  • Other conventional chewing gum additives known to one having ordinary skill in the chewing gum art may also be used in die chewing gum base.
  • the gum composition may include effective amounts of conventional additives selected from the group consisting of sweetening agents (sweeteners), plasticizers, softeners, emulsifiers, waxes, fillers, bulking agents, mineral adjuvants, flavoring agents (flavors, flavorings), coloring agents (colorants, colorings), antioxidants, acidulants, thickeners, mixtures thereof and the like.
  • sweeteners e.g., sorbitol or other sugar alcohol or mixtures thereof
  • the sugar sweetener can also function as a bulking agent.
  • the plasticizers, softeners, mineral adjuvants, colorants, waxes and antioxidants discussed above as being suitable for use in the gum base may also be used in the gum composition.
  • examples of other conventional additives which may be used include emulsifiers, such as lecithin and glyceryl monostearate, thickeners, used alone or in combination with other softeners, such as methyl cellulose, alginates, ca ⁇ ageenan, xanthan gum, gelatin, carob, tragacanth, locust bean, and carboxy methyl cellulose, acidulants such as malic acid, adipic acid, citric acid, tartaric acid, fumaric acid, and mixtures thereof, and fillers, such as those discussed above under die category of mineral adjuvants.
  • the fillers when used may be utilized in an amount up to about 60%, by weight of the gum composition.
  • Bulking agents (carriers, extenders) suitable for use in chewing gums include sweetening agents selected from the group consisting of monosaccharides, disaccharides, poly-saccharides, sugar alcohols, and mixtures thereof; polydextrose; maltodextrins; minerals, such as calcium carbonate, talc, titanium dioxide, dicalcium phosphate, and die like. Bulking agents may be used in amounts up to about 90%, by weight of the final gum composition, with amounts from about 40% to about 70%, by weight of the gum composition being prefe ⁇ ed , with from about 50% to about 65%, by weight, being more prefe ⁇ ed and from about 55% to about 60%, by weight of the chewing gum composition, being most preferred.
  • the sweetening agent used may be selected from a wide range of materials including water-soluble sweeteners, water-soluble artificial sweeteners, water- soluble sweeteners derived from naturally occurring water-soluble sweeteners, dipeptide based sweeteners, and protein based sweeteners, including mixtures thereof.
  • sweeteners representative categories and examples include: (a) water-soluble sweetening agents such as monosaccharides, disaccharides and polysaccharides such as xylose, ribulose, glucose (dextrose), mannose, galactose, fructose (levulose), sucrose (sugar), maltose, invert sugar (a mixture of fructose and glucose derived from sucrose), partially hydrolyzed starch, co syrup solids, dihydrochalcones, monellin, steviosides, glycyrrhizin, and sugar alcohols such as sorbitol, mannitol, maltitol, hydrogenated starch hydrolysates and mixtures thereof; (b) water-soluble artificial sweeteners such as soluble saccharin salts, i.e., sodium or calcium saccharin salts, cyclamate salts, the sodium, ammonium or calcium saltof3,4-dihydro-6-methyl-l;
  • water-soluble sweeteners derived from naturally occurring water- soluble sweeteners such as chlorinated derivatives of ordinary sugar (sucrose), known, for example, under die product designation of Sucralose; and
  • an effective amount of sweetener is utilized to provide die level of bulk and or sweetness desired, and this amount will vary with the sweetener selected.
  • This amount of sweetener will normally be present in amounts from about 0.0025% to about 90%, by weight of the gum composition, depending upon die sweetener used.
  • the exact range of amounts for each type of sweetener is well known in the art and is not the subject of the present invention.
  • the amount of sweetener ordinarily necessary to achieve the desired level of sweetness is independent from die flavor level achieved from flavor oils.
  • Prefe ⁇ ed sugar based-sweeteners are sugar (sucrose), com syrup and mixtures thereof.
  • Prefe ⁇ ed sugarless sweeteners are the sugar alcohols, artificial sweeteners, dipeptide based sweeteners and mixtures thereof.
  • sugar alcohols are used in die sugarless compositions because these sweeteners can be used in amounts which are sufficient to provide bulk as well as the desired level of sweetness.
  • Prefe ⁇ ed sugar alcohols are selected from the group consisting of sorbitol, xylitol, maltitol, mannitol, and mixtures thereof. More preferably, sorbitol or a mixture of sorbitol and mannitol is utilized. The gamma form of sorbitol is preferred.
  • An artificial sweetener or dipeptide based sweetener is preferably added to the gum compositions which contain sugar alcohols.
  • the coloring agents useful in the gum compositions are used in amounts effective to produce the desired color. These coloring agents include pigments which may be incorporated in amounts up to about 6% by weight of the gum composition. A prefe ⁇ ed pigment, titanium dioxide, may be incorporated in amounts up to about 2%, and preferably less than about 1% by weight of the composition.
  • the colorants may also include natural food colors and dyes suitable for food, drug and cosmetic applications. These colorants are known as F.D.& C. dyes and lakes.
  • the materials acceptable for the foregoing uses are preferably water-soluble. Illustrative nonlimiting examples include die indigoid dye known as F.D.& C. Blue No.2, which is the disodium salt of 5,5-indigotindisulfonic acid.
  • the dye known as F.D.& C. Green No.l comprises a triphenylmethane dye and is the monosodium salt of 4-[4-(N- ethyl-p-sulfoniumbe-izyl--mino)liphenylmethylene]-[l-(N-ethyl-N-p-sulfoniumbenzyl)- delta-2,5-cyclohexadieneimine].
  • a full recitation of all F.D.& C. colorants and their co ⁇ esponding chemical structures may be found in the Kirk-Othmer Encyclopedia of Chemical Technology. 3rd Edition, in volume 5 at pages 857-884, which text is incorporated herein by reference.
  • Suitable oils and fats usable in gum compositions include partially hydrogenated vegetable or animal fats, such as coconut oil, palm kernel oil, beef tallow, lard, and the like. These ingredients when used are generally present in amounts up to about 7%, by weight, and preferably up to about 3.5%, by weight of the gum composition.
  • therapeutically effective amounts of the therapeutic wound healing compositions of the present invention may be admixed into a chewing gum. These amounts are readily determined by tiiose skilled in the art without the need for undue experimentation.
  • die final chewing gum composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 10% and a chewing gum composition in a quantity sufficient to bring die total amount of composition to 100%, by weight of the chewing gum composition.
  • die final chewing gum composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most preferred embodiment, the final chewing gum composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and a chewing gum composition in a quantity sufficient to bring the total amount of composition to 100%, by weight of the chewing gum composition.
  • the present invention extends to metiiods of making the therapeutic chewing gum compositions.
  • the therapeutic wound healing compositions may be incorporated into an otiierwise conventional chewing gum composition using standard techniques and equipment known to those skilled in the art.
  • the apparatus useful in accordance with the present invention comprises mixing and heating apparatus well known in the chewing gum manufacturing arts, and therefore the selection of the specific apparatus will be apparent to the artisan.
  • a gum base is heated to a temperature sufficiently high enough to soften the base without adversely effecting the physical and chemical make up of the base.
  • the optimum temperatures utilized may vary depending upon die composition of the gum base used, but such temperatures are readily determined by tiiose skilled in the art without undue experimentation.
  • the gum base is conventionally melted at temperatures that range from about 60°C. to about 120°C. for a period of time sufficient to render the base molten.
  • the gum base may be heated under these conditions for a period of about thirty minutes just prior to being admixed incrementally with the remaining ingredients of the base such as the plasticizer, fillers, the bulking agent and/or sweeteners, die softener and coloring agents to plasticize the blend as well as to modulate die hardness, viscoelasticity and formability of the base.
  • the chewing gum base is then blended witii the therapeutic wound healing composition of the present invention which may have been previously blended witii other traditional ingredients. Mixing is continued until a uniform mixture of gum composition is obtained. Thereafter the gum composition mixture may be formed into desirable chewing gum shapes.
  • die invention is directed to a therapeutic pharmaceutical composition for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises:
  • Embodiment One selected from the group consisting of:
  • (IA) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • LB (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; and
  • fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells
  • LC a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells
  • LC a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells
  • LC (a) an antioxidant
  • LD lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof;
  • the pharmaceutically acceptable carrier may be selected from die group consisting of pharmaceutical appliances, topical vehicles, and ingestible vehicle.
  • the invention is directed to a method for preparing a therapeutic pharmaceutical composition for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of :
  • Embodiment One selected from the group consisting of:
  • LB (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; and
  • lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof;
  • step (C) admixing the therapeutic wound healing composition from step (A) and die pharmaceutically acceptable carrier from step (B) to form a therapeutic pharmaceutical composition.
  • This study demonstrates a comparison of the viability of U937 monocytic cells after exposure of the cells to various antioxidants and combinations of antioxidants. This study also demonstrate a comparison of the levels of hydrogen peroxide produced by U937 monocytic cells and mammalian epidermal keratinocytes after exposure of the cells to various antioxidants and combinations of antioxidants.
  • Mammalian epidermal keratinocytes and monocytes were employed to examine the ability of various antioxidants to reduce levels of hydrogen peroxide in tiiese cells. Hydrogen peroxide was measured after die cells were exposed to ultraviolet light in the wavelength range from 290 to 320 nm (UV-B) or to the inflammatory compound 12-0-tetradecanoyl-phorbol-13-acetate (TPA). Sodium pyruvate was tested at various concentrations to determine the effect of concentrations of this antioxidant on the hydrogen peroxide production by epidermal cells and monocytes.
  • Magnesium pyruvate, calcium pyruvate, zinc pyruvate, and combinations of sodium pyruvate with ascorbic acid, lactic acid, and Vitamin E were then tested to determine the effect of these salts and combinations of antioxidants on the hydrogen peroxide production by epidermal cells and monocytes.
  • Mammalian epidermal keratinocytes were isolated by trypsin-zation of epithelial sheets and grown in modified basal MCDB 153 medium supplemented witii epidermal growth factor, bovine pituitary extract, and hydrocortisone. Cells were maintained in a humidified incubator witii 5% carbon dioxide at 37°C. Keratinocytes were seeded in 60 mm culture dishes at a cell density of 3 x 10 cells per dish and die cultures were exposed to 1 M.E.D. dose of ultraviolet-B light (100 mJ/cm ) or treated with lOO ng/ml of TPA.
  • U937 monocytic cells are a cultured cell line grown in RPMI media with 10% fetal calf serum. Cells were maintained in a 60 mm culture dish at 5% carbon dioxide at 37°C. at a seeding density not exceeding 1 x 10 6 cells per dish.
  • Sodium pyruvate, lactic acid, ascorbic acid, and Vitamin E were dissolved in distilled water, with sufficient surfactant.
  • the concentrations of the sodium pyruvate solutions prepared were 1 mM, 10 mM, 50 mM, 100 mM, and 200 mM.
  • the concentrations of the lactic acid solutions prepared were 1.0%, 0.1%, and 0.05%.
  • the concentrations of the ascorbic acid solutions prepared were 1.0%, 0.1%, 0.05%, and 0.025%.
  • the concentrations of the Vitamin E solutions prepared were 1 U, 10 U, 50 U, and 100 U.
  • the test solutions were adjusted to a pH value of 7.4 with 1.0N sodium hydroxide solution and then sterile filtered.
  • test solution or combination of test solutions was added to die cells immediately prior to exposure of the cells to ultraviolet light-B or TPA [100ng/ml].
  • Stock solutions were prepared so tiiat the vehicle did not constitute more than 1% of the total volume of the culture media.
  • DCFH-DA dichlorofluorescein diacetate
  • DCFH-DA is a non-polar non- fluorescent compound that readily diffuses into cells where it is hydrolyzed to the polar non-fluorescent derivative DCFH which then becomes trapped within the cells.
  • DCFH is oxidized to die highly fluorescent compound DCF.
  • cellular fluorescence intensity is directly proportional to the level of intracellular hydrogen peroxide produced. Cellular fluorescence intensity can be monitored by fluorimetry and by flow cytometry.
  • Figure 2 depicts in bar graph format the viability of U937 monocytic cells after exposure of the cells to various combinations of antioxidants.
  • the viability of U937 monocytic cells was measured after exposure to no antioxidant (Example 6, control), to ascorbic acid and lactic acid (Example 7), to ascorbic acid and Vitamin E (Example 8), to sodium pyruvate and ascorbic acid (Example 9), to sodium pyruvate and lactic acid (Example 10), to sodium pyruvate and Vitamin E (Example 11), to lactic acid and Vitamin E (Example 12), and to sodium pyruvate, ascorbic acid, and lactic acid (Example 13).
  • Figure 1 shows that ascorbic acid is cytotoxic to monocytes at concentrations as low as 0.25%.
  • Figure 2 shows that the cytotoxicity of ascorbic acid was reversed by the addition of 10 mM of sodium pyruvate.
  • Figures 1 and 2 show that the viability rate of 15% to 20% of the cells when treated with ascorbic acid was increased to 95% to 98% upon addition of sodium pyruvate. Lactic acid and
  • Vitamin E did not reverse the cytotoxicity of ascorbic acid.
  • Sodium pyruvate was then tested at various concentrations to determine die effect of concentrations of this antioxidant on the hydrogen peroxide production by epidermal cells and monocytes.
  • Mammalian epidermal keratinocytes and monocytes were exposed to (a) 1 M.E.D. dose of ultraviolet light-B and (b) 100 ng/ml of 12-O- tetradecanoylphorbol-13-acetate (TPA) in the presence of sodium pyruvate at the following concentrations: 200 mM, 100 mM, 50 mM, 10 mM, 1 mM.
  • the optimum concentration of sodium pyruvate to reduce the hydrogen peroxide production by epidermal cells and monocytes was found to be 10 mM. Concentrations of sodium pyruvate of 50 mM and above were cytotoxic to both epidermal keratinocytes and monocytes.
  • Magnesium pyruvate, calcium pyruvate, zinc pyruvate, ascorbic acid, lactic acid, and Vitamin E, and combinations of sodium pyruvate with ascorbic acid, lactic acid, and Vitamin E were then tested to determine the effect of these salts and combinations of antioxidants on die hydrogen peroxide production by epidermal cells and monocytes.
  • the following test solutions were prepared.
  • the optimum concentration of lactic acid to reduce the hydrogen peroxide production by epidermal cells and monocytes was found to be 0.05%.
  • the optimum concentration of ascorbic acid was found to be 0.025%.
  • the higher concentrations of both of these compounds were found to be cytotoxic to both types of cells.
  • the optimum concentration of Vitamin E was found to be 50 U.
  • Figure 3 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells after exposure of the cells to no antioxidant
  • Example 14 control
  • sodium pyruvate Example 15
  • ascorbic acid Example 16
  • lactic acid Example 17
  • Vitamin E Example 18
  • Sodium pyruvate and Vitamin E significantly reduced die hydrogen peroxide production by monocytes.
  • Figure 4 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells after exposure of the cells to various combinations of antioxidants. Specifically, the levels of hydrogen peroxide produced by U937 monocytic cells were measured after exposure to no antioxidant (Example 19, control), to ascorbic acid and lactic acid (Example 20), to ascorbic acid and Vitamin E
  • Example 21 to sodium pyruvate and ascorbic acid (Example 22), to sodium pyruvate and lactic acid (Example 23), to sodium pyruvate and Vitamin E (Example 24), to lactic acid and Vitamin E (Example 25), and to sodium pyruvate, ascorbic acid, and lactic acid (Example 26).
  • the combination of lactic acid (0.05%) and Vitamin E (50 U) significantly reduced the hydrogen peroxide production by monocytes.
  • the morphological alterations in epidermal keratinocytes were observed in control cultures and in cultures exposed to ultraviolet-B. Cells in the layer closest to the dermis are basal keratinocytes.
  • the differentiation pattern results in cells enucleating and forming comified envelopes at the uppermost portion of the epidermis, the statum corneum.
  • the differentiation of keratinocytes is controlled by the levels of calcium, magnesium, and otiier elements in the medium. Cells in culture systems promoting differentiation appear as an epidermal sheet forming attachments or tight junctions with each other. Keratinocytes that become nonadherent or float in the media were considered responding to a cytotoxic event.
  • Ascorbic Acid Cells were floating in a cytotoxic response to ascorbic acid.
  • Lactic Acid Cells appeared dramatically altered as an epidermal sheet and as flat granular cells.
  • Vitamin E Cells appeared the same as the cells in the control culture.
  • 50 U Vitamin E and 10 mM Sodium Pyruvate Cells increased in number and changed in appearance resembling the cells in the sodium pyruvate culture.
  • Ascorbic Acid Cells were nonadherent and floating in a cytotoxic response to ascorbic acid greater than the cytotoxic response of the co ⁇ esponding cells without ultraviolet-B light exposure.
  • Lactic Acid Cells formed an epidermal sheet and were more granular than cells in the control culture without ultraviolet-B light exposure.
  • Vitamin E Cell growth was inhibited but cells appeared similar to cells in the control culture without ultraviolet-B light exposure.
  • Vitamin E 50 U Vitamin E and 10 mM Sodium Pvmvate: Cells appeared similar to cells in the control culture and proliferated to a greater extent than cells in the control cultures without ultraviolet-B light exposure.
  • Vitamin E at 50 U and 100 U.
  • Mammalian epidermal keratinocytes and U937 monocytic cells and the test solutions of sodium pyruvate, lactic acid, ascorbic acid, and Vitamin E were prepared as describe above for Examples 1-26. Intracellular hydrogen peroxide production by the mammalian epidermal keratinocytes and U937 monocytes was also measured as described above.
  • a mixture of fatty acids derived from chicken fat was prepared for addition to the cultured cells by mixing 0.1% of the chicken fat with the culture media. At the temperature of the culture media, 37°C, the chicken fat was miscible. This chicken fat mixture was added to cultures of cells prior to exposure of the cells to ultraviolet-B light or TPA treatment.
  • mammalian epidermal keratinocytes and monocytes were exposed to (a) 1 M.E.D. dose of ultraviolet light-B and (b) 100 ng/ml of 12-O-tetradecanoylphorbol-13-acetate in the presence of various antioxidants and combinations of antioxidants with and witiiout a mixture of saturated and unsaturated fatty acids [0.1%, 0.5%, and 1.0% chicken fat].
  • Figure 5 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells after exposure of the cells to various combinations of antioxidants with and without a mixture of saturated and unsaturated fatty acids. Specifically, die levels of hydrogen peroxide produced by U937 monocytic cells were measured after exposure to lactic acid and Vitamin E without fatty acids (Example 27) and with fatty acids (Example 28), to ascorbic acid and lactic acid without fatty acids (Example 29) and witii fatty acids (Example 30), and to ascorbic acid and Vitamin E witiiout fatty acids (Example 31) and witii fatty acids (Example 32).
  • Figure 6 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal keratinocytes after exposure of the cells to various antioxidants with and witiiout a mixture of saturated and unsaturated fatty acids.
  • the levels of hydrogen peroxide produced by epidermal keratinocytes were measured after exposure to no antioxidant witiiout fatty acids (Example 33, control) and witii fatty acids (Example 34), to sodium pyruvate without fatty acids (Example 35) and witii fatty acids (Example 36), to ascorbic acid witiiout fatty acids (Example 37) and witii fatty acids (Example 38), to lactic acid without fatty acids (Example 39) and witii fatty acids (Example 40), and to Vitamin E without fatty acids (Example 41) and witii fatty acids (Example 42).
  • FIG. 7 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal keratinocytes after exposure of the cells to various combinations of antioxidants with and witiiout a mixture of saturated and unsaturated fatty acids.
  • die levels of hydrogen peroxide produced by epidermal keratinocytes were measured after exposure to no antioxidant witiiout fatty acids (Example 43, control) and witii fatty acids (Example 44), to sodium pyruvate and ascorbic acid without fatty acids (Example 45) and witii fatty acids (Example 46), to sodium pyruvate and lactic acid witiiout fatty acids (Example 47) and witii fatty acids (Example 48), to sodium pyruvate and Vitamin E witiiout fatty acids (Example 49) and with fatty acids (Example 50), and to ascorbic acid and Vitamin E without fatty acids (Example 51) and witii fatty acids (Example 52).
  • Human epidermal keratinocytes were isolated by trypsinization of epithelial sheets and grown in modified base MCDB 153 medium supplemented witii epidermal growth factor and bovine pituitary extract. Cells were seeded in culture dishes at a density of 3 x 10 5 /dish. Prior to exposure to UV B light (100mJ/cm 2 ) or treatment with lOOng/ml TPA, die cultures were treated with the appropriate concentration of wound healing components. Intracellular production of hydrogen peroxide was measured using DCFH-DA, a nonpolar compound that readily diffuses into cells, hydrolyzed to a nonpolar derivative. In the presence of intracellular hydrogen peroxide, DCFH is oxidized to a highly fluorescent compound DCF. Thus, cellular fluorescence intensity is directly proportional to levels of hydrogen peroxide produced and can be monitored by flow cytometry. Hydrogen peroxide is cytotoxic, therefore lower levels of hydrogen peroxide production is desirable for cellular viability.
  • Column 1 shows the different treatment groups.
  • Column 2 shows the production of H 2 0 2 in control cells (fmol/cell).
  • Column 3 shows the production of H 0 after treatment with wound healing components.
  • Column 4 shows the difference in production of H 2 0 2 from control after the treatment.
  • the wound healing compositions of Examples A-D were prepared having the compositions set out in Table A.
  • Wound healing composition A was commercially available Preparation H .
  • Wound healing composition B was a petrolatum base formulation containing live yeast cell derivative, shark oil, and a mixture of sodium pyruvate, vitamin E, and chicken fat.
  • Wound healing composition C was a petrolatum base formulation containing live yeast cell derivative and shark oil.
  • Wound healing composition D was a petrolatum base formulation only.
  • the animals were sacrificed on day 3 and day 7 using cervical dislocation.
  • the dorsal skin including the incision was dissected without the subcutaneous tissue.
  • the skin was placed in neutral buffered formalin and subsequently sectioned and stained with hematoxylin and eosin.
  • the wounds were examined microscopically and representative tissue sections were photographed.
  • Figures 9 and 10 show tiiat Formulation B, which was a petrolatum base formulation containing live yeast cell derivative, shark oil, and a mixture of sodium pyruvate, vitamin E, and chicken fat, was a significantly better wound healing agent than the other formulations. These results are supported by die subjective grading of die wound closures and die speed of healing on each day (1-7) of the experiment as well as on the objective histological examination of tissue sections to measure the extent of inflammatory cell infiltrate within the wound and die extent of epithelialization at the wound edges. The final result was that less scar tissue was present at day 7 on the mice treated witii Formulation B.
  • Formulation D which was a white petrolatum formulation only, was judged to be significantly more effective to promote healing than either Formulation C, which was a petrolatum base formulation containing shark liver oil and live yeast cell derivative, or Formulation A, which was Preparation H .
  • the superior ability of Formulation D over Formulation C to improve healing may result from a delay in die healing process caused when the live yeast cell derivative is depleted and die cells shift to an alternative nutrient source.
  • the presence of the mixture of sodium pyruvate, vitamin E, and chicken fat in Formulation B apparently offsets the depletion of the live yeast cell derivative.
  • Formulation C which was a petrolatum base formulation containing live yeast cell derivative and shark oil, was judged comparable to the control (untreated wound) in speed of wound closure and extent of healing.
  • Formulation A which was Preparation HTM, appeared to be die least effective healing formulation by both subjective grading of wound healing and by objective examination of tissue sections.
  • the superior ability of Formulation D and Formulation C over Formulation A to improve healing may be due to their ability to act as an occlusive wound dressing tiiat prevents transepidermal water loss and thus promotes healing and wound closure.
  • the poor ability of Formulation A to improve healing may be due to the potential cytotoxicity of phenylmercuric nitrate present in Preparation H as a preservative.
  • the wound healing composition (LA) comprises (a) pyruvate, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids.
  • Sunscreen agents can help prevent sunburn by screening ultra violet light but do not heal injured mammalian cells.
  • Anti- inflammatory agents can reduce inflammation (erythema) in a patient but do not promote the wound healing process.
  • Wound healing compositions can increase the resuscitation rate of injured mammalian cells and die proliferation rate of new mammalian cells to replace dead cells. Wound healing compositions can also minimize oxygen radical damage from ultra violet light. Applicants have found tiiat the combination of a sunscreen agent, an anti-inflammatory agent, and a wound healing composition results in a therapeutic sunscreen-wound healing compositions useful for minimizing and treating sunburn damage.
  • the sunscreen- wound healing compositions may optionally contain a therapeutically effective amount of a topical anesthetic to further reduce the duration and severity of sunburn.
  • the combination of the sunscreen agent, the anti-inflammatory agent, and die wound healing compositions of the present invention provides a pharmaceutical composition useful for minimizing and treating sunburn damage and having an enhanced ability to prevent and reduce injury to mammalian cells and further increase the resuscitation rate of injured mammalian cells.
  • the tissue damage associated witii sunburn is believed to be caused by die production of cellular produced active oxygen species.
  • Combination of the sunscreen agent, anti-inflammatory agent, and die wound healing compositions helps suppress such reactive oxygen-linked tissue injury.
  • Sunscreen agents are compounds which provide broad spectrum protection from ultra violet A and ultra violet B light from die sun.
  • the sunscreen agents in the sunscreen-wound healing compositions of the present invention may be selected from a wide range of tiierapeutic agents and mixtures of therapeutic agents.
  • Nonlimiting illustrative specific examples of sunscreen agents include etiiylhexyl p- methoxycinnamate, octyl methoxycinnamate, octyl dimethyl »-aminobenzoic acid, 2- ethylhexyl salicylate, octyl salicylate, menthyl anthranilate, octocrylene, padimate o, titanium dioxide, urea, and oxybenzone.
  • the sunscreen agent is oxybenzone.
  • the amount of sunscreen agent used in die present invention is a therapeutically effective amount and may vary depending upon the therapeutic dosage recommended or permitted for the particular sunscreen agent. In general, the amount of sunscreen agent present is the ordinary dosage required to obtain the desired result. Such dosages are known to the skilled practitioner in the medical arts and are not a part of the present invention.
  • the sunscreen agent in the sunscreen-wound healing composition is present in an amount from about 1 % to about 30%, preferably from about 2% to about 25%, and more preferably from about 4% to about 20%, by weight.
  • Anti-inflammatory agents are compounds that counteract or suppress the inflammatory process.
  • the anti-inflammatory agents in the sunscreen-wound healing compositions of the present invention may be selected from a wide variety of steroidal, non-steroidal, and salicylate water-soluble and water-insoluble drugs and their acid addition or metallic salts. Both organic and inorganic salts may be used provided the anti-inflammatory agent maintains its medicament value.
  • the anti-inflammatory agents may be selected from a wide range of therapeutic agents and mixtures of therapeutic agents which may be administered in sustained release or prolonged action form.
  • Nonlimiting illustrative specific examples of non-steroidal anti-inflammatory agents include the following medicaments: ibuprofen, naproxen, sulindac, diflunisal, piroxicam, indomethacin, etodolac, meclofenamate sodium, fenoproben calcium, ketoprofen, mefenamic acid, nabumetone, ketorolac tromethamine, diclofenac, and evening primrose oil (containing about 72% linoleic acid and about 9% gamma- linolenic acid).
  • Nonlimiting illustrative specific examples of salicylate anti- inflammatory agents include die following medicaments: acetylsalicylic acid, mesalamine, salsalate, diflunisal, salicylsalicylic acid, and choline magnesium trisalicylate.
  • Nonlimiting illustrative specific examples of steroidal anti-inflammatory agents include die following medicaments: flunisolide, triamcinoline, triamcinoline acetonide, beclometiiasone diproprionate, betamethasone diproprionate, hydrocortisone, cortisone, dexamethasone, predinisone, methyl prednisolone, and prednisolone.
  • Prefe ⁇ ed anti-inflammatory agents to be employed may be selected from the group consisting of ibuprofen, naproxen, sulindac, diflunisal, piroxicam, indomethacin, etodolac, meclofenamate sodium, fenoproben calcium, ketoprofen, mefenamic acid, nabumetone, ketorolac tromethamine, diclofenac, evening primrose oil, acetylsalicylic acid, mesalamine, salsalate, diflunisal, salicylsalicylic acid, choline magnesium trisalicylate, flunisolide, triamcinoline, triamcinoline acetonide, beclometiiasone diproprionate, betamethasone diproprionate, hydrocortisone, cortisone, dexamethasone, predinisone, methyl prednisolone, and prednisolone.
  • die anti-inflammatory agent is selected from the group consisting of ibuprofen, naproxen, sulindac, diflunisal, piroxicam, indomethacin, etodolac, meclofenamate sodium, fenoproben calcium, ketoprofen, mefenamic acid, nabumetone, ketorolac tromethamine, diclofenac, and evening primrose oil.
  • the - ti-infl-ur-matory agent is evening primrose oil.
  • the anti-inflammatory agent of the present invention may be used in many distinct physical forms well known in the pharmaceutical art to provide an initial dosage of the anti-inflammatory agent and/or a further time-release form of the anti- inflammatory agent. Without being limited thereto, such physical forms include free forms and encapsulated forms, and mixtures thereof.
  • the amount of anti-inflammatory agent used in die present invention is a therapeutically effective amount and may vary depending upon the therapeutic dosage recommended or permitted for the particular anti-inflammatory agent. In general, the amount of anti-inflammatory agent present is the ordinary dosage required to obtain the desired result. Such dosages are known to the skilled practitioner in the medical arts and are not a part of the present invention.
  • the anti- inflammatory agent in the sunscreen-wound healing composition is present in an amount from about 0.01% to about 10%, preferably from about 0.1% to about 5%, and more preferably from about 1% to about 3%, by weight.
  • the therapeutic sunscreen-wound healing compositions of the present invention further comprise a topical anesthetic agent.
  • Anesthetic agents are compounds that induce the loss of tactile sensibility and die sensation of pain.
  • the anesthetic agents in the sunscreen-wound healing compositions of the present invention may be selected from a wide range of therapeutic agents and mixtures of therapeutic agents.
  • Nonlimiting illustrative specific examples of topical anesthetic agents include pramoxine hydrochloride, lidocaine, and benzocaine.
  • the amount of anesthetic agent used in the present invention is a therapeutically effective amount and may vary depending upon die tiierapeutic dosage recommended or permitted for the particular anesthetic agents. In general, the amount of anesthetic agents present is the ordinary dosage required to obtain the desired result. Such dosages are known to the skilled practitioner in the medical arts and are not a part of the present invention.
  • the anesthetic agent in the sunscreen-wound healing composition is present in an amount from about 1 % to about 30%, preferably from about 2% to about 25%, and more preferably from about 2.5% to about 20%, by weight.
  • a therapeutic sunscreen-wound healing composition is made by forming an admixture of the wound healing components of Embodiment One (LA-D), a sunscreen agent, and an anti- inflammatory agent.
  • a sunscreen- wound healing tiierapeutic composition is made by forming an admixture of a sunscreen agent, an anti-inflammatory agent, and a wound healing composition comprising (a) a pyruvate, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids.
  • a sunscreen-wound healing therapeutic composition is made by forming an admixture of a sunscreen agent, an anti-inflammatory agent, and a wound healing composition comprising (a) a pyruvate, (b) a lactate, and (c) a mixture of saturated and unsaturated fatty acids.
  • a sunscreen-wound healing therapeutic composition is made by forming an admixture of a sunscreen agent, an anti-inflammatory agent, and a wound healing composition comprising (a) an antioxidant, and (b) a mixture of saturated and unsaturated fatty acids.
  • a sunscreen-wound healing therapeutic composition is made by forming an admixture of admixture of a sunscreen agent, an anti-inflammatory agent, and a wound healing composition comprising (a) a lactate, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids.
  • die invention is directed to a method for preparing a therapeutic sunscreen-wound healing composition (IA + X) useful to minimize and treat sunburn damage which comprises the steps of admixing a therapeutically effective amount of the following ingredients:
  • pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • the present invention extends to metiiods for employing the therapeutic sunscreen-wound healing compositions (LA-D + X).
  • a therapeutic composition is employed by contacting the therapeutic composition with the skin to be exposed to the sun.
  • die invention is directed to a method for minimizing and treating sunburn in a human with a sunscreen-wound healing composition (LA + X) which comprises the steps of:
  • pyruvate selected from die group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • the therapeutic sunscreen- wound healing compositions (LA-D + X) of the present invention may be further combined with medicaments useful for treating wounds (M) to form augmented sunscreen-wound healing compositions (I.A-D + X + M).
  • augmented sunscreen-wound healing compositions I.A-D + X + M.
  • die combination of the sunscreen-wound healing composition of the present invention and the medicament useful for treating wounds provides an augmented sunscreen-wound healing composition having an enhanced ability to increase the proliferation and resuscitation rate of mammalian cells.
  • the therapeutic compositions of the present invention may be used in combination with medicaments useful for treating wounds such as immunostimulating agents (Betafectin ), antiviral agents, antikeratolytic agents, anti-inflammatory agents, antifungal agents, tretinoin, other sunscreen agents, dermatological agents, topical antihistamine agents, antibacterial agents, bioadhesive agents, respiratory bursting inhibitors (lactic acid, adenosine), inhibitors of prostaglandin synthesis (ibuprofen, aspirin, indomethacin, meclofenomic acid, retinoic acid, padimate O, meclomen, oxybenzone), steroidal anti-inflammatory agents (corticosteroids including synthetic analogs), antimicrobial agents (neosporin ointment, silvadine), antiseptic agents, anesthetic agents (pramoxine hydrochloride, lidocaine, benzocaine), cell nutrient media, bu relief medications, sun bum medications, acne preparations
  • the medicament useful for treating wounds is selected from the group consisting of immunostimulating agents, antiviral agents, antikeratolytic agents, anti-inflammatory agents, antifungal agents, tretinoin, sunscreen agents, dermatological agents, topical antihistamine agents, antibacterial agents, bioadhesive agents, respiratory bursting inhibitors, inhibitors of prostaglandin synthesis, antimicrobial agents, cell nutrient media, scar reducing agents, and mixtures thereof.
  • the medicament useful for treating wounds is selected from die group consisting of immunostimulating agents, antiviral agents, antikeratolytic agents, anti-inflammatory agents, antifungal agents, acne treating agents, sunscreen agents, dermatological agents, antihistamine agents, antibacterial agents, bioadhesive agents, and mixtures thereof.
  • the invention is directed to an augmented sunscreen-wound healing composition (IA + X + M) useful to minimize and treat sunburn damage which comprises:
  • composition which comprises a therapeutically effective amount of:
  • wound healing composition comprises:
  • pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • an antioxidant an antioxidant
  • the present invention extends to methods for making the augmented sunscreen-wound healing compositions.
  • the augmented compositions are made by admixing the therapeutic sunscreen-wound healing composition with the medicament useful for treating wounds to prepare the augmented sunscreen-wound healing composition.
  • the present invention also extends to metiiods for employing the augmented sunscreen-wound healing compositions.
  • an augmented sunscreen-wound healing composition is employed by contacting the composition with the skin to be exposed to the sun.
  • the invention is directed to a method for minimizing and treating sunburn in a human with an augmented sunscreen-wound healing composition (LA + X + M) which comprises the steps of: (A) providing a therapeutically effective amount of a sunscreen-wound healing composition which comprises:
  • wound healing composition comprises:
  • pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • the types of wounds which may be healed using the sunscreen-wound healing compositions and the augmented sunscreen-wound healing compositions of the present invention are inflammation wounds induced by sunburn.
  • the therapeutic compositions may be used topically to protect and accelerate the healing of injured tissue.
  • Methods for treating sunburn comprise topically administering the compositions of the present invention directly to the skin of the human prior to exposure to the sun.
  • the composition is maintained in contact with the skin for a period of time sufficient to increase the proliferation and resuscitation rate of the cells.
  • die inventive therapeutic sunscreen-wound healing compositions and augmented sunscreen-wound healing compositions may be stored for future use or may be formulated in effective amounts with pharmaceutically acceptable carriers such as pharmaceutical appliances and topical vehicles to prepare a wide variety of pharmaceutical compositions.
  • pharmaceutically acceptable carriers which may be employed and die metiiods used to prepare the pharmaceutical compositions have been described above in connection witii the formulations of the wound healing compositions of Embodiment One (LA-D).
  • die invention is directed to a sunscreen- wound healing pharmaceutical composition which comprises:
  • composition (A) a therapeutic sunscreen-wound healing composition (LA + X) which comprises:
  • wound healing composition comprises:
  • pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • a pharmaceutically acceptable carrier selected from the group consisting of pharmaceutical appliances, bioadhesives, and occlusive vehicles.
  • the invention is directed to a method for preparing a pharmaceutical composition for increasing the proliferation and resuscitation rate of mammalian cells, which comprises the steps of: (A) providing a therapeutically effective amount of a sunscreen-wound healing composition (LA + X) which comprises:
  • pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
  • step (C) admixing the sunscreen-wound healing composition from step (A) and die pharmaceutically acceptable carrier from step (B) to form a pharmaceutical composition.
  • both the cream and lotion test products provided low level sun protection (SPF values less then 2) and minor improvements in UV-induced erythema as compared witii i ⁇ adiated control sites not treated with test products.
  • test material(s) A sample of each test material(s) was reserved and stored for a period of five (5) years. At the conclusion of the clinical study, the remaining test material(s) was discarded. All information regarding the receipt, storage and disposition of the material(s) was also recorded on a Clinical Material Record form. All test materials were kept in a locked product storage room accessible to clinical staff members only.
  • Product (1) was LubridermTM Lotion containing 2% sodium pyruvate, 1% vitamin E, and 1% chicken fat.
  • Product (2) was LubridermTM Cream containing aquaphor/petrolatum cream with 10% sodium pyruvate, 5% vitamin E, and 5% chicken fat.
  • a Xenon Arc Solar Simulator 150W was used which has a continuous emission spectrum in the UVA and UVB range (290 to 400 nanometers). Less than 1 % of its total energy was composed of "non-solar" wavelengths below 290 nm. The output was monitored daily, using the Robert-Berge meter. Additional measurements of the energy output were taken and recorded as needed.
  • MED. Minimal Erythema Dose - The time of light exposure necessary to produce a minimal perceptible erythema (redness) on the skin, discernible sixteen (16) to twenty four (24) hours later.
  • SPF Sun Protection Factor - The ultraviolet energy required to produce an MED on protected skin, divided by die ultraviolet energy required to produce an MED on unprotected skin.
  • Minimal Sun Protection Product - Sunscreen products tiiat provide an SPF value of 2 to 4, and offer the least protection from sunbuming, but permit suntanning.
  • Moderate Sun Protection Product - Sunscreen products tiiat provide an SPF value of 4 to 6, and offer moderate protection from sunbuming, and permit some suntanning.
  • Extra Sun Protection Product - Sunscreen products that provide an SPF value of 6 to 8, and offer extra protection from sunbuming, and permit limited suntanning.
  • Maximal Sun Protection Product - Sunscreen products that provide an SPF value of 8 to under 15, and offer maximal protection from sunbuming, and permit little or no suntanning.
  • Ultra Sun Protection Product - Sunscreen products tiiat provide an SPF value of 15 or greater, and offer the most protection from sunbuming and permit no suntanning.
  • MED is defined as die time of light exposure required to produce a minimal perceptible erythema reaction discernible sixteen (16) to twenty-four (24) hours after irradiation using a standardized ultraviolet light source that emits UVB(290- 320 nm) as all or part of its emission spectrum.
  • Subjects who were candidates for this testing have skin types commonly referred to as Fitzpatrick skin types, of Category I, EL, or in according to the following definitions:
  • Category I always bu s easily, never tans.
  • Category II Always bums easily, tans minimally.
  • the anticipated MED was estimated from skin type (I, ⁇ , or HI) of the individual and die irradiation size calculated based on energy output of the Xenon lamp.
  • the (5) sites were irradiated on the basis of a geometric progression of 1.25, i.e., each site receiving 25% more exposure than the site to its left.
  • Sixteen (16) to twenty-four (24) hours later, the MED was determined by establishing the site which exhibited die least amount of perceptible erythema.
  • test material was applied as follows:
  • a 50 cm area was marked on the back of the subject. Then, a uniform application of 0.1 g or ml or product was applied to die area with careful spreading and gende but thorough rubbing over the entire marked area. The product was permitted to dry for fifteen (15) to thirty (30) minutes.
  • the irradiation for site exposure was again calculated on the basis of a geometric progression of 1.25 with the site to the right receiving more irradiation tiian the site to its left.
  • the sites were then exposed to ultraviolet light from the solar simulator.
  • the procedure was repeated using the control product, an 8% homosalate standard.
  • test sites All sites were covered, but not occluded, witii a gauze dressing. Application of die product to the same sites was repeated approximately six hours later and the following mornings and afternoons for four consecutive days. Evaluation of the test sites was made each afternoon prior to product application by a trained clinical evaluator who had no participation in any other aspect of the study procedure. A final evaluation was made on die morning of Day 8, the last product application having occurred on die previous Friday (Day 4). Evaluations of each test site were made according to die following scale:
  • the cream product showed improvement over the control site in two subjects for all four comparison intervals, in one subject at two of the four intervals, and two other subjects at one of the four intervals.
  • the lotion product showed improvement in one subject at all four intervals, in one subject at three intervals, in two subjects at two intervals, and, in one subject at one interval.
  • eight of thirteen subjects showed no differences at any of die four intervals in comparisons between the test product sites and the control sites for both the cream and die lotion products. While the effects noted here must again be stressed as minimal, it is, nonetheless, noteworthy that improvements, where noted, favored effects of the test products over the control and not the opposite: twenty-four instances of possible positive effects from the two test products compared to one such possible instance at the control site.

Abstract

The present invention pertains to therapeutic sunscreen-wound healing compositions useful to minimize and treat sunburn damage. The compositions comprise a therapeutically effective amount of (1) a sunscreen agent; (2) an anti-inflammatory; and, (3) a wound healing composition. In one embodiment the wound healing composition comprises (a) pyruvate; (b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids. The therapeutic sunscreen-wound healing compositions may be utilized in a wide variety of pharmaceutical products. This invention also relates to methods for preparing and using the therapeutic sunscreen-wound healing compositions and the pharmaceutical products in which the therapeutic compositions may be used.

Description

SUNSCREEN-WOUND HEALING COMPOSITION
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention pertains to therapeutic sunscreen-wound healing compositions useful to minimize and treat sunburn damage. More particularly, the sunscreen-wound healing compositions comprise a sunscreen agent, an anti-inflammatory agent, and a therapeutic wound healing composition and/or its metabolites. This invention also pertains to methods for preparing and using the therapeutic sunscreen-wound healing compositions and the pharmaceutical products in which the therapeutic compositions may be used.
A preferred embodiment of the therapeutic wound healing composition of this invention comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. 2. Description of the Background Wound Healing
Wounds are internal or external bodily injuries or lesions caused by physical means, such as mechanical, chemical viral, bacterial, or thermal means, which disrupt the normal continuity of structures. Such bodily injuries include contusions, wounds in which the skin is unbroken, incisions, wounds in which the skin is broken by a cutting instrument, and lacerations, wounds in which the skin is broken by a dull or blunt instrument. Wounds may be caused by accidents or by surgical procedures. Patients who suffer major wounds could benefit from an enhancement in the wound healing process.
Wound healing consists of a series of processes whereby injured tissue is repaired, specialized tissue is regenerated, and new tissue is reorganized. Wound healing consists of three major phases: a) an inflammation phase (0-3 days), b) a cellular proliferation phase (3-12 days), and (c) a remodeling phase (3 days-6 months).
During the inflammation phase, platelet aggregation and clotting form a matrix which traps plasma proteins and blood cells to induce the influx of various types of ceils.
During the cellular proliferation phase, new connective or granulation tissue and blood vessels are formed. During the remodeling phase, granulation tissue is replaced by a network of collagen and elastin fibers leading to the formation of scar tissue.
When cells are injured or killed as a result of a wound, a wound healing step is desirable to resuscitate the injured cells and produce new cells to replace the dead cells. The healing process requires the reversal of cytotoxicity, the suppression of inflammation, and the stimulation of cellular viability and proliferation. Wounds require low levels of oxygen in the initial stages of healing to suppress oxidative damage and higher levels of oxygen in the later stages of healing to promote collagen formation by fibroblasts. Mammalian cells are continuously exposed to activated oxygen species such as superoxide (O2-), hydrogen peroxide (H2O2), hydroxyl radical (OH ), and singlet oxygen ( O2). In vivo, these reactive oxygen intermediates are generated by cells in response to aerobic metabolism, catabolism of drugs and other xenobiotics, ultraviolet and x-ray radiation, and the respiratory burst of phagocytic cells (such as white blood cells) to kill invading bacteria such as those introduced through wounds. Hydrogen peroxide, for example, is produced during respiration of most living organisms especially by stressed and injured cells.
These active oxygen species can injure cells. An important example of such damage is lipid peroxidation which involves the oxidative degradation of unsaturated lipids. Lipid peroxidation is highly detrimental to membrane structure and function and can cause numerous cytopathological effects. Cells defend against lipid peroxidation by producing radical scavengers such as superoxide dismutase, catalase, and peroxidase. Injured cells have a decreased ability to produce radical scavengers.
Excess hydrogen peroxide can react with DNA to cause backbone breakage, produce mutations, and alter and liberate bases. Hydrogen peroxide can also react with pyrimidines to open the 5, 6-double bond, which reaction inhibits the ability of pyrimidines to hydrogen bond to complementary bases, Hallaender et al. (1971). Such oxidative biochemical injury can result in the loss of cellular membrane integrity, reduced enzyme activity, changes in transport kinetics, changes in membrane lipid content, and leakage of potassium ions, amino acids, and other cellular material.
Antioxidants have been shown to inhibit damage associated with active oxygen species. For example, pyruvate and other ---//.λα-ketoacids have been reported to react rapidly and stoichiometrically with hydrogen peroxide to protect cells from cytolytic effects, O'Donnell-Tormey et al., J. Exp. Med., 165, pp. 500-514 (1987).
United States Patents Nos. 3,920,835, 3,984,556, and 3,988,470, all issued to Van Scott et al., disclose methods for treating acne, dandruff, and palmar keratosis, respectively, which consist of applying to the affected area a topical composition comprising from about 1% to about 20% of a lower aliphatic compound containing from two to six carbon atoms selected from the group consisting of Alpha- hydroxyacids, -*-/ . Λα-ketoacids and esters thereof, and 3-hydroxybutryic acid in a phaπnaceutically acceptable carrier. The aliphatic compounds include pyruvic acid and lactic acid.
United States Patents Nos. 4,105,783 and 4,197,316, both issued to Yu et al., disclose a method and composition, respectively, for treating dry skin which consists of applying to the affected area a topical composition comprising from about 1 % to about 20% of a compound selected from the group consisting of amides and ammonium salts of Alp Aα-hydroxyacids, jS-hydroxyacids, and -4//>Λα-ketoacids in a pharmaceutically acceptable carrier. The compounds include the amides and ammonium salts of pyruvic acid and lactic acid.
United States Patent No. 4,234,599, issued to Van Scott et al., discloses a method for treating actinic and nonactinic skin keratoses which consists of applying to the affected area a topical composition comprising an effective amount of a compound selected from the group consisting of -4//.Λα-hydroxyacids, jS-hydroxyacids, and -4//.Λ-.-ketoacids in a pharmaceutically acceptable carrier. The acidic compounds include pyruvic acid and lactic acid.
United States Patent No. 4,294,852, issued to Wildnauer et al., discloses a composition for treating skin which comprises the Alpha-hydroxyacids.^-hydroxyacids, and /pΛα-ketoacids disclosed above by Van Scott et al. in combination with C^-Cg aliphatic alcohols.
United States Patent No. 4,663,166, issued to Veech, discloses an electrolyte solution which comprises a mixture of L-lactate and pyruvate in a ratio from 20: 1 to 1:1, respectively, or a mixture of D-^-hydroxybutyrate and acetoacetate, in a ratio from 6:1 to 0.5:1, respectively.
Sodium pyruvate has been reported to reduce the number of erosions, ulcers, and hemorrhages on the gastric mucosa in guinea pigs and rats caused by acetylsalicylic acid. The analgesic and antipyretic properties of acetylsalicylic acid were not impaired by sodium pyruvate, Puschmann, Arzneimittelforschung, 33, pp. 410-415 and 415-416 (1983).
Pyruvate has been reported to exert a positive inotropic effect in stunned myocardium, which is a prolonged ventricular dysfunction following brief periods of coronary artery occlusions which does not produce irreversible damage, Mentzer et al., Ann. Surg., 209, pp. 629-633 (1989).
Pyruvate has been reported to produce a relative stabilization of left ventricular pressure and work parameter and to reduce the size of infarctions. Pyruvate improves resumption of spontaneous beating of the heart and restoration of normal rates and pressure development, Bunger et al, J. Mol. Cell. Cardiol., 18, pp. 423*438 (1986), Mochizuki et al, J. Physiol. (Paris), 76, pp. 805-812 (1980), Regitz et al, Cardiovasc. Res., 15, pp. 652-658 (1981), Giannelli et al, Ann. Thorac. Surg., 21, pp. 386-396 (1976).
Sodium pyruvate has been reported to act as an antagonist to cyanide intoxication (presumably through the formation of a cyanohydrin) and to protect against the lethal effects of sodium sulfide and to retard the onset and development of functional, morphological, and biochemical measures of acrylamide neuropathy of axons, Schwartz et α/., Toxicol. Appl. Pharmacol., 50, pp. 437-442 (1979), Sabri et al, Brain Res., 483, pp. 1-11 (1989).
A chemotherapeutic cure of advanced L1210 leukemia has been reported using sodium pyruvate to restore abnormally deformed red blood cells to normal. The deformed red blood cells prevented adequate drug delivery to tumor cells, Cohen, Cancer Chemother. Pharmacol., 5, pp. 175-179 (1981).
Primary cultures of heterotopic tracheal transplant exposed in vivo to 7, 12- dimethyl-benz(a)anthracene were reported to be successfully maintained in enrichment medium supplemented with sodium pyruvate along with cultures of interleukin-2 stimulated peripheral blood lymphocytes, and plasmacytomas and hybridomas, pig embryos, and human blastocysts, Shacter, J. Immunol. Methods, 99, pp. 259-270 (1987), Marchok et al, Cancer Res., 37, pp. 1811-1821 (1977), Davis, J. Reprod. Fertil. Suppl., 33, pp. 115-124 (1985), Okamoto et al, No To Shinkei, 38, pp. 593-598 (1986), Cohen et al, J. In Vitro Fert. Embryo Transfer, 2, pp. 59-64
(1985).
United States Patents Nos. 4,158,057, 4,351,835, 4,415,576, and 4,645,764, all issued to Stanko, disclose methods for preventing the accumulation of fat in the liver of a mammal due to the ingestion of alcohol, for controlling weight in a mammal, for inhibiting body fat while increasing protein concentration in a mammal, and for controlling the deposition of body fat in a living being, respectively. The methods comprise administering to the mammal a therapeutic mixture of pyruvate and dihydroxyacetone, and optionally riboflavin. United States Patent No. 4,548,937, issued to Stanko, discloses a method for controlling the weight gain of a mammal which comprises administering to the mammal a therapeutically effective amount of pyruvate, and optionally riboflavin. United States Patent No. 4,812,479, issued to Stanko, discloses a method for controlling the weight gain of a mammal which comprises administering to the mammal a therapeutically effective amount of dihydroxyacetone, and optionally riboflavin and pyruvate.
Rats fed a calcium-oxalate lithogenic diet including sodium pyruvate were reported to develop fewer urinary calculi (stones) than control rats not given sodium pyruvate, Ogawa et al, Hinyokika Kiyo, 32, pp. 1341-1347 (1986).
United States Patent No. 4,521,375, issued to Houlsby, discloses a method for sterilizing surfaces which come into contact with living tissue. The method comprises sterilizing the surface with aqueous hydrogen peroxide and then neutralizing the surface with pyruvic acid. United States Patent No. 4,416,982, issued to Tauda et al, discloses a method for decomposing hydrogen peroxide by reacting the hydrogen peroxide with a phenol or aniline derivative in the presence of peroxidase.
United States Patent No. 4,696,917, issued to Lindstrom et al, discloses an eye irrigation solution which comprises Eagle's Minimum Essential Medium with Earle's salts, chondroitin sulfate, a buffer solution, 2-mercaptoethanol, and a pyruvate. The irrigation solution may optionally contain ascorbic acid and ---/pΛα-tocopherol. United States Patent No. 4,725,586, issued to Lindstrom et al, discloses an irrigation solution which comprises a balanced salt solution, chondroitin sulfate, a buffer solution,
2-mercaptoethanol. sodium bicarbonate or dextrose, a pyruvate, a sodium phosphate buffer system, and cystine. The irrigation solution may optionally contain ascorbic acid mXnd mma-xocc herol.
lilted States Patent No. 3,887,702 issued to Baldwin, discloses a composition for treating fingernails and toenails which consists essentially of soybean oil or sunflower oil in combination with Vitamin E.
United States Patent No. 4,847,069, issued to Bissett et al, discloses a photoprotective composition comprising (a) a sorbohydroxamic acid, (b) an anti- inflammatory agent selected from steroidal anti-inflammatory agents and a natural anti- inflammatory agent, and (c) a topical carrier. Fatty acids may be present as an emollient. United States Patent No. 4,847,071, issued to Bissett et al, discloses a photoprotective composition comprising (a) a tocopherol or tocopherol ester radical scavenger, (b) an anti-inflammatory agent selected from steroidal anti-inflammatory agents and a natural anti-inflammatory agent, and (c) a topical carrier. United States Patent No. 4,847,072, issued to Bissett et al, discloses a topical composition comprising not more than 25% tocopherol sorbate in a topical carrier.
United States Patent No. 4,533,637, issued to Yamane et al, discloses a culture medium which comprises a carbon source, a nucleic acid source precursor, amino acids, vitamins, minerals, a lipophilic nutrient, and serum albumin, and cyclodextrins. The lipophilic substances include unsaturated fatty acids and lipophilic vitamins such as Vitamin A, D, and E. Ascorbic acid may also be present.
United Kingdom patent application no. 2,196,348A, to Kovar et al, discloses a synthetic culture medium which comprises inorganic salts, monosaccharides, amino acids, vitamins, buffering agents, and optionally sodium pyruvate adding magnesium hydroxide or magnesium oxide to the emulsion. The oil phase may include chicken fat.
United States Patent No. 4,284,630, issued to Yu et al, discloses a method for stabilizing a water-in-oil emulsion which comprises adding magnesium hydroxide or magnesium oxide to the emulsion. The oil phase may include chicken fat.
Preparation H has been reported to increase the rate of wound healing in artificially created rectal ulcers. The active ingredients in Preparation H are skin respiratory factor and shark liver oil, Subramanyam et al, Digestive Diseases and Sciences, 29, pp. 829-832 (1984).
The addition of sodium pyruvate to bacterial and yeast systems has been reported to inhibit hydrogen peroxide production, enhance growth, and protect the systems against the toxicity of reactive oxygen intermediates. The unsaturated fatty acids and saturated fatty acids contained within chicken fat enhanced membrane repair and reduced cytotoxicity. The antioxidants glutathione and thioglycollate reduced the injury induced by oxygen radical species, Martin, Ph.D. thesis, (1987-89).
United States Patent No. 4,615,697, issued to Robinson, discloses a controlled release treatment composition comprising a treating agent and a bioadhesive agent comprising a water-swellable but water-insoluble, fibrous cross-linked carboxy- fiinctional polymer.
European patent application no. 0410696A1, to Kellaway et al, discloses a mucoadhesive delivery system comprising a treating agent and a polyacrylic acid cross- linked with from about 1% to about 20% by weight of a polyhydroxy compound such as a sugar, cyclitol, or lower polyhydric alcohol.
Inflammation
Inflammation is a nonspecific response caused by several kinds of injury, including penetration of the host by infectious agents. The distinguishing feature of inflammation is dilation and increased permeability of minute blood vessels. Direct injury, such as that caused by toxins elaborated by microorganisms, leads to destruction of vascular endothelium and increased permeability to plasma proteins, especially in the venules and venular capillaries. Mediators of secondary injury are liberated from the site of direct injury. As a result, gaps form between vascular endothelial cells through which plasma proteins escape. Granulocytes, monocytes, and erythrocytes may also leave vascular channels. Mediators of secondary injury include unknown substances and histamine, peptides (kinins), kinin-forming enzymes (kininogenases), and a globulin permeability factor. These mediators are blocked from action by antihistamines and sympathoamines, and are most pronounced in effect on venules, although lymphvascular endothelium also becomes more porous as a part of secondary injury.
The beneficial effect of the inflammatory response is the production of: (1) leukocytes in great numbers; (2) plasma proteins, nonspecific and specific humoral agents, fibrinogen that on conversion to fibrin aids in localization of the infectious process while acting as a matrix for phagocytosis; and (3) increased blood and lymph flow that dilutes and flushes toxic materials while causing a local increase in temperature.
In the early stages of inflammation, the exudate is alkaline and neutrophilic polymorphonuclear leukocytes predominate. As lactic acid accumulates, presumably from glycolysis, the Ph drops and macrophages become the predominant cell type.
Lactic acid and antibodies in the inflammatory exudate may inhibit parasites, but the major anti-infectious effect of the inflammatory response is attributable to phagocytic cells.
The inflammatory response consists of three successive phases: (a) increased vascular permeability with resulting edema and swelling, (b) cellular infiltration and phagocytoses, and (c) proliferation of the fibroblasts, synthesizing new connective tissue to repair the injury. A large number of so-called mediators of inflammation have been implicated in the inflammatory process primarily in terms of their capacity to induce vasodilation and increase permeability.
The initial increase in capillary permeability and vasodilation in an inflamed joint is followed by an increase in metabolism of the joint tissues. Leakage of fibrinogen into the wound, where proteolytic enzymes convert it into fibrin, establishes a capillary and lymphatic blockade. The concentrations of components of the ground substance of connective tissue collagen, mucopolysaccharides, glycoproteins, and nonfibrous proteins are greatly increased during this process. As the exudative phase of the inflammation subsides, the fibroblast is found to be the dominant cell in the wounded zone. It first proliferates, then synthesizes extracellular material, including new collagen fibers and acid mucopolysaccharides, which are laid down to form the new tissue matrix.
The inflammatory phenomenon includes fenestration of the microvasculature, leakage of the elements of blood into the interstitial spaces, and migration of leukocytes into the inflamed tissue. On a macroscopic level, this is usually accompanied by the familiar clinical signs of erythema, edema tenderness
(hyperalgesia), and pain. During this complex response, chemical mediators such as histamine, 5-hydroxytryptamine (5-HT), slow-reacting substance of anaphylaxis (SRS-A), various chemotactic factors, bradykinin, and prostaglandins are liberated locally. Phagocytic cells migrate into the area, and cellular lysosomal membranes may be ruptured, releasing lytic enzymes. All these events may contribute to the inflammatory response. Sunburn
Sunburn is an acute reaction by the skin to excessive exposure to sunlight. Ordinary sunburn results from overexposure of the skin to ultraviolet waves of about 3000 Angstroms. Sunburn symptoms appear in 1 to 24 hours and, except in severe reactions, pass their peak in 72 hours. Following exposure to sunlight, the epidermis thickens and the melanocytes produce melanin at an increased rate, providing some natural protection against further exposure. Skin changes range from a mild erythema with subsequent evanescent scaling, to pain, swelling, skin tenderness, and blisters from more prolonged exposure. Fever, chills, weakness, and shock may appear if a large portion of the body surface is affected. Secondary infection, particularly furunculosis, is the most common late complication. The skin may remain hypervulnerable to sunlight for one to several weeks when pronounced exfoliation has occurred.
Sunscreen agents are very effective for preventing sunburn. A useful sunscreen agent formulation is 5% .αr -aminobenzoic acid (PABA) in ethyl alcohol or in a gel. The esters of αrα-aminobenzoic acid in an alcohol base are only slightly less effective. Patients who do not tolerate αra-aminobenzoic acid or its esters may use a benzophenone or titanium dioxide sunscreen agent. Early treatment of extensive and severe sunburn with a topical or systemic corticosteroid decreases discomfort considerably.
Chronic exposure to sunlight has an aging effect on the skin. Wrinkling and elastosis (yellow discoloration with small yellow nodules) are the most common consequences of long-term exposure. Precancerous keratotic lesions (actinic keratoses) are a frequent, disturbing consequence of years of overexposure. Squamous and basal cell carcinoma of the skin are especially common in people who work outdoors. SUMMARY OF THE INVENTION
This invention pertains to therapeutic sunscreen-wound healing compositions useful to minimize and treat sunburn damage. The compositions of this invention comprise a therapeutically effective amount of (1) a sunscreen agent; (2) an anti- inflammatory agent; and, (3) a wound healing composition. A preferred embodiment of the wound healing composition of this invention comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. The therapeutic sunscreen-wound healing compositions of this invention may be utilized in a wide variety of pharmaceutical products. This invention also relates to methods for preparing and using the therapeutic sunscreen-wound healing compositions and the pharmaceutical products in which the therapeutic compositions may be used.
This invention further comprises augmented therapeutic sunscreen-wound healing compositions comprising (1) a sunscreen agent; (2) an anti-inflammatory agent; and, (3) a therapeutic wound healing composition in combination with one or more additional medicaments. This invention also relates to methods for preparing and using the augmented therapeutic antikeratolytic-wound healing compositions and the pharmaceutical products in which the augmented compositions may be used.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 depicts in bar graph format the viability of U937 monocytic cells following exposure of the cells to various antioxidants (Examples 1-5).
Figure 2 depicts in bar graph format the viability of U937 monocytic cells following exposure of the cells to various combinations of antioxidants (Examples 6- 13). Figure 3 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells following exposure of the cells to various antioxidants (Examples 14-18).
Figure 4 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells following exposure of the cells to various combinations of antioxidants (Examples 19-26).
Figure 5 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells following exposure of the cells to various combinations of antioxidants with and without a mixture of saturated and unsaturated fatty acids (Examples 27-32).
Figure 6 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal keratinocytes following exposure of the cells to various antioxidants with and without a mixture of saturated and unsaturated fatty acids (Examples 33-42).
Figure 7 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal keratinocytes following exposure of the cells to various combinations of antioxidants with and without a mixture of saturated and unsaturated fatty acids (Examples 43-52).
Figure 8 depicts in bar graph format a summary analysis of the levels of hydrogen peroxide produced by epidermal keratinocytes following exposure of the cells to the individual components of the wound healing composition, to various combinations of the wound healing composition, and to the wound healing composition.
Figure 9 is a photograph of wounded mice after 4 days of treatment with:
Preparation H (Example A); a petrolatum base formulation containing live yeast cell derivative, shark oil, and a mixture of sodium pyruvate, vitamin E, and chicken fat (Example B); a petrolatum base formulation containing live yeast cell derivative and shark oil (Example C); and no composition (Example E, control).
Figure 10 is a photograph of a wounded mouse after 4 days of treatment with a petrolatum base formulation only (Example D).
DETAILED DESCRIPTION OF THE INVENTION
This invention pertains to therapeutic sunscreen-wound healing compositions which comprise (1) a sunscreen agent; (2) an anti-inflammatory; and, (3) a wound healing composition and/or its metabolites. In one embodiment the wound healing composition comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
Applicant has discovered therapeutic wound healing compositions for preventing and reducing injury to mammalian cells and increasing the resuscitation rate of injured mammalian cells. Cells treated with the therapeutic wound healing compositions of the present invention show decreased levels of hydrogen peroxide production, increased resistance to cytotoxic agents, increased rates of proliferation, and increased viability. Cellular cultures containing the therapeutic wound healing compositions showed enhanced differentiation and proliferation over control cultures and rapidly formed attachments or tight junctions between the cells to form an epidermal sheet. Wounded mammals treated with the therapeutic wound healing compositions show significantly improved wound closing and healing over untreated mammals and mammals treated with conventional healing compositions. The wound healing compositions may be used alone or in combination with other medicaments. The therapeutic wound healing compositions of this invention are described as Embodiment One. There are several aspects of Embodiment One. In a first aspect, (LA), the therapeutic wound healing composition comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. In a second aspect of Embodiment One (LB), the therapeutic wound healing composition comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. In a third aspect of Embodiment One (LC), the therapeutic wound healing composition comprises (a) an antioxidant and (b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. In a fourth aspect of Embodiment One (I.D), the therapeutic wound healing composition comprises (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
The therapeutic wound healing compositions of this invention are further combined with a therapeutically effective amount of (1) a sunscreen agent and (2) an anti-inflammatory (collectively referred to as "X") to form therapeutic sunscreen-wound healing compositions (I.A-D + X). The therapeutic sunscreen-wound healing compositions may be used alone or in combination with other medicaments. This invention also pertains to methods for preparing and using the sunscreen- wound healing compositions and the pharmaceutical products in which the therapeutic compositions may be used. The therapeutic sunscreen-wound healing compositions of this invention may be further combined with one or more additional medicaments for treating wounds to form augmented sunscreen-wound healing compositions. This invention also relates to methods for preparing and using the augmented therapeutic sunscreen-wound healing compositions and the pharmaceutical products in which the augmented compositions may be used.
The term "injured cell" as used herein means a cell that has any activity disrupted for any reason. For example, an injured cell may be a cell that has injured membranes or damaged DNA, RNA, and ribosomes, for example, a cell which has (a) injured membranes so that transport through the membranes is diminished resulting in an increase in toxins and normal cellular wastes inside the cell and a decrease in nutrients and other components necessary for cellular repair inside the cell, (b) an increase in concentration of oxygen radicals inside the cell because of the decreased ability of the cell to produce antioxidants and enzymes, or (c) damaged DNA, RNA, and ribosomes which must be repaired or replaced before normal cellular functions can be resumed. The term "resuscitation" of injured mammalian cells as used herein means the reversal of cytotoxicity, the stabilization of the cellular membrane, an increase in the proliferation rate of the cell, and or the normalization of cellular functions such as the secretion of growth factors, hormones, and the like. The term "cytotoxicity" as used herein means a condition caused by a cytotoxic agent that injures the cell. Injured cells do not proliferate because injured cells expend all energy on cellular repair. Aiding cellular repair promotes cellular proliferation.
The term "prodrug", as used herein, refers to compounds which undergo biotransformation prior to exhibiting their pharmacological effects. The chemical modification of drugs to overcome pharmaceutical problems has also been termed "drug latentiation." Drug latentiation is the chemical modification of a biologically active compound to form a new compound which upon in vivo enzymatic attack will liberate the parent compound. The chemical alterations of the parent compound are such that the change in physicochemical properties will affect the absorption, distribution and enzymatic metabolism. The definition of drug latentiation has also been extended to include nonenzymatic regeneration of the parent compound. Regeneration takes place as a consequence of hydrolytic, dissociative, and other reactions not necessarily enzyme mediated. The terms prodrugs, latentiated drugs, and bioreversible derivatives are used interchangeably. By inference, latentiation implies a time lag element or time component involved in regenerating the bioactive parent molecule in vivo. The term prodrug is general in that it includes latentiated drug derivatives as well as those substances which are converted after administration to the actual substance which combines with receptors. The term prodrug is a generic term for agents which undergo biotransformation prior to exhibiting their pharmacological actions. In the case where the administered drug is not the active agent, but rather is biotransfoimed to the active agent, the term "prodrug" also includes compounds which may not necessarily undergo biotransformation to the administered drug but may undergo biotransformation to the active agent which exhibits the desired pharmacological effect.
The term "metabolite", as used herein, refers to any substance produced by metabolism or by a metabolic process. "Metabolism", as used herein, refers to the various chemical reactions involved in the transformation of molecules or chemical compounds occurring in tissue and the cells therein.
I. Wound Healing Compositions A. Embodiment One (I.A-D) The cells which may be treated with the therapeutic wound healing compositions in the present invention are mammalian cells. Although applicant will describe the present therapeutic wound healing compositions as useful for treating mammalian epidermal keratinocytes and mammalian monocytes, applicant contemplates that the therapeutic wound healing compositions may be used to protect or resuscitate all mammalian cells. Keratinocytes are representative of normal mammalian cells and are the fastest proliferating cells in the body. The correlation between the reaction of keratinocytes to injury and therapy and that of mammalian cells in general is very high. Monocytes are representative of specialized mammalian cells such as the white blood cells in the immune system and the organ cells in liver, kidney, heart, and brain. The mammalian cells may be treated in vivo and in vitro.
Epidermal keratinocytes are the specialized epithelial cells of the epidermis which synthesize keratin, a scleroprotein which is the principal constituent of epidermis, hair, nails, homy tissue, and the organic matrix of the enamel of teeth. Mammalian epidermal keratinocytes constitute about 95% of the epidermal cells and together with melanocytes form the binary system of the epidermis. In its various successive stages, epidermal keratinocytes are also known as basal cells, prickle cells, and granular cells.
Monocytes are mononuclear phagocytic leukocytes which undergo respiratory bursting and are involved in reactive oxygen mediated damage within the epidermis. Leukocytes are white blood cells or corpuscles which may be classified into two main groups: granular leukocytes (granulocytes) which are leukocytes with abundant granules in the cytoplasm and nongranular leukocytes (nongranulocytes) which are leukocytes without specific granules in the cytoplasm and which include the lymphocytes and monocytes. Phagocyte cells are cells which ingest microorganisms or other cells and foreign particles. Monocytes are also known as large mononuclear leukocytes, and hyaline or transitional leukocytes.
Epidermal keratinocytic cells and monocytic cells have multiple oxygen generating mechanisms and the degree to which each type of mechanism functions differs in each type of cell. In monocytes, for example, the respiratory bursting process is more pronounced than in epidermal keratinocytes. Hence, the components in the therapeutic wound healing compositions of the present invention may vary depending upon the types of cells involved in the condition being treated.
As set out above, in a first aspect of Embodiment One (LA), the therapeutic wound healing composition for treating mammalian cells, preferably epidermal keratinocytes, comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. In a second aspect of Embodiment One (LB), the therapeutic wound healing composition for treating mammalian cells, preferably epidermal keratinocytes, comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. In a third aspect of Embodiment One (LC), the therapeutic wound healing composition for treating mammalian cells, preferably epidermal keratinocytes, comprises (a) an antioxidant and (b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. In a fourth aspect of Embodiment One (I.D), the therapeutic wound healing composition for treating mammalian cells, preferably monocytes, comprises (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
Pyruvic acid (2-oxopropanoic acid^/pA -ketopropionic acid, CH3COCOOH) or pyruvate is a fundamental intermediate in protein and carbohydrate metabolism and in the citric acid cycle. The citric acid cycle (tricarboxylic acid cycle, Kreb's cycle) is the major reaction sequence which executes the reduction of oxygen to generate adenosine triphosphate (ATP) by oxidizing organic compounds in respiring tissues to provide electrons to the transport system. Acetyl coenzyme A ("active acetyl") is oxidized in this process and is thereafter utilized in a variety of biological processes and is a precursor in the biosynthesis of many fatty acids and sterols. The two major sources of acetyl coenzyme A are derived from the metabolism of glucose and fatty acids. Glycolysis consists of a series of transformations wherein each glucose molecule is transformed in the cellular cytoplasm into two molecules of pyruvic acid. Pyruvic acid may then enter the mitochondria where it is oxidized by coenzyme A in the presence of enzymes and cofactors to acetyl coenzyme A. Acetyl coenzyme A can then enter the citric acid cycle.
In muscle, pyruvic acid (derived from glycogen) can be reduced to lactic acid during anerobic metabolism which can occur during exercise. Lactic acid is reoxidized and partially retransformed to glycogen during rest. Pyruvate can also act as an antioxidant to neutralize oxygen radicals in the cell and can be used in the multifunction oxidase system to reverse cytotoxicity.
The pyruvate in the present invention may be selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, prodrugs of pyruvic acid, and mixtures thereof. In general, the pharmaceutically acceptable salts of pyruvic acid may be alkali salts and alkaline earth salts. Preferably, the pyruvate is selected from the group consisting of pyruvic acid, lithium pyruvate, sodium pyruvate, potassium pyruvate, magnesium pyruvate, calcium pyruvate, zinc pyruvate, manganese pyruvate, methyl pyruvate, Alpha-ketoglutaric acid, and mixtures thereof. More preferably, the pyruvate is selected from the group of salts consisting of sodium pyruvate, potassium pyruvate, magnesium pyruvate, calcium pyruvate, zinc pyruvate, manganese pyruvate, and the like, and mixtures thereof. Most preferably, the pyruvate is sodium pyruvate.
The amount of pyruvate present in the therapeutic wound healing compositions of the present invention is a therapeutically effective amount. A therapeutically effective amount of pyruvate is that amount of pyruvate necessary for the inventive composition to prevent and reduce injury to mammalian cells or increase the resuscitation rate of injured mammalian cells. The exact amount of pyruvate is a matter of preference subject to such factors as the type of condition being treated as well as the other ingredients in the composition. In a preferred embodiment, pyruvate is present in the therapeutic wound healing composition in an amount from about 10% to about 50%, preferably from about 20% to about 45%, and more preferably from about 25% to about 40%, by weight of the therapeutic wound healing composition.
/--Lactic acid ((S)-2-hydroxypropanoic acid, (+) -- λα-hydroxypropionic acid, CH3CHOHCOOH) or lactate occurs in small quantities in the blood and muscle fluid of mammals. Lactic acid concentration increases in muscle and blood after vigorous activity. Lactate is a component in the cellular feedback mechanism and inhibits the natural respiratory bursting process of cells thereby suppressing the production of oxygen radicals.
The lactate in the present invention may be selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, prodrugs of lactic acid, and mixtures thereof. In general, the pharmaceutically acceptable salts of lactic acid may be alkali salts and alkaline earth salts. Preferably, the lactate is selected from the group consisting of lactic acid, lithium lactate, sodium lactate, potassium lactate, magnesium lactate, calcium lactate, zinc lactate, manganese lactate, and the like, and mixtures thereof. More preferably, the lactate is selected from the group consisting of lactic acid, sodium lactate, potassium lactate, magnesium lactate, calcium lactate, zinc lactate, manganese lactate, and mixtures thereof. Most preferably, the lactate is lactic acid.
The amount of lactate present in the therapeutic wound healing compositions of the present invention is a therapeutically effective amount. A therapeutically effective amount of lactate is that amount of lactate necessary for the inventive composition to prevent and reduce injury to mammalian cells or increase the resuscitation rate of injured mammalian cells. For an ingestible composition, a therapeutically effective amount of lactate is that amount necessary to suppress the respiratory bursting process of white blood cells to protect and resuscitate the mammalian cells. In general, a therapeutically effective amount of lactate in an ingestible composition is from about 5 to about 10 times the amount of lactate normally found in serum. The exact amount of lactate is a matter of preference subject to such factors as the type of condition being treated as well as the other ingredients in the composition. In a preferred embodiment, lactate is present in the therapeutic wound healing composition in an amount from about 10% to about 50%, preferably from about 20% to about 45%, and more preferably from about 25% to about 40%, by weight of the therapeutic wound healing composition.
Antioxidants are substances which inhibit oxidation or suppress reactions promoted by oxygen or peroxides. Antioxidants, especially lipid-soluble antioxidants, can be absorbed into the cellular membrane to neutralize oxygen radicals and thereby protect the membrane. The antioxidants useful in the present invention may be selected from the group consisting of all forms of Vitamin A (retinol), all forms of
Vitamin2 (3, 4-didehydroretinol), all forms of carotene such as ---//. Aα-carotene, β- carotene (beta, .-carotene), gα/πmα-carotene, de/tø-carotene, all forms of Vitamin C (D-ascorbic acid, L-ascorbic acid), all forms of tocopherol such as Vitamin E (Alpha- tocopherol, 3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltri-decyl)-2H-l- benzopyran-6-ol), /.-tocopherol, gαmmα-tocopherol, rfe/tα-tocopherol, tocoquinone, tocotrienol, and Vitamin E esters which readily undergo hydrolysis to Vitamin E such as Vitamin E acetate and Vitamin E succinate, and pharmaceutically acceptable Vitamin E salts such as Vitamin E phosphate, prodrugs of Vitamin A, carotene, Vitamin C, and Vitamin E, pharmaceutically acceptable salts of Vitamin A, carotene, Vitamin C, and Vitamin E, and the like, and mixtures thereof. Preferably, the antioxidant is selected from the group of lipid-soluble antioxidants consisting of Vitamin A, jβ-carotene, Vitamin E, Vitamin E acetate, and mixtures thereof. More preferably, the antioxidant is Vitamin E or Vitamin E acetate. Most preferably, the antioxidant is Vitamin E acetate.
The amount of antioxidant present in the therapeutic wound healing compositions of the present invention is a therapeutically effective amount. A therapeutically effective amount of antioxidant is that amount of antioxidant necessary for the inventive composition to prevent and reduce injury to mammalian cells or increase the resuscitation rate of injured mammalian cells. The exact amount of antioxidant is a matter of preference subject to such factors as the type of condition being treated as well as the other ingredients in the composition. In a prefeπed embodiment, the antioxidant is present in the therapeutic wound healing composition in an amount from about 0.1% to about 40%, preferably from about 0.2% to about 30%, and more preferably from about 0.5% to about 20%, by weight of the therapeutic wound healing composition.
The mixture of saturated and unsaturated fatty acids in the present invention are those fatty acids required for the repair of mammalian cellular membranes and the production of new cells. Fatty acids are carboxylic acid compounds found in animal and vegetable fat and oil. Fatty acids are classified as lipids and are composed of chains of alkyl groups containing from 4 to 22 carbon atoms and 0-3 double bonds and characterized by a terminal carboxyl group, -COOH. Fatty acids may be saturated or unsaturated and may be solid, semisolid, or liquid. The most common saturated fatty acids are butyric acid (C4), lauric acid (C^), palmitic acid (Cjg), and stearic acid (Cjg). Unsaturated fatty acids are usually derived from vegetables and consist of alkyl chains containing from 16 to 22 carbon atoms and 0-3 double bonds with the characteristic terminal carboxyl group. The most common unsaturated fatty acids are oleic acid, linoleic acid, and linolenic acid (all Cjo acids).
In general, the mixture of saturated and unsaturated fatty acids required for the repair of mammalian cellular membranes in the present invention may be derived from animal and vegetable fats and waxes, prodrugs of saturated and unsaturated fatty acids useful in the present invention, and mixtures thereof. For example, the fatty acids in the therapeutic wound healing composition may be in the form of mono-, di-, or trigylcerides, or free fatty acids, or mixtures thereof, which are readily available for the repair of injured cells. Cells produce the chemical components and the energy required for cellular viability and store excess energy in the form of fat. Fat is adipose tissue stored between organs of the body to furnish a reserve supply of energy. The preferred animal fats and waxes have a fatty acid composition similar to that of human fat and the fat contained in human breast milk. The prefeπed animal fats and waxes may be selected from the group consisting of human fat, chicken fat, cow fat (defined herein as a bovine domestic animal regardless of sex or age), sheep fat, horse fat, pig fat, and whale fat. The more prefeπed animal fats and waxes may be selected from the group consisting of human fat and chicken fat. The most prefeπed animal fat is human fat. Mixtures of other fats and waxes, such as vegetable waxes (especially sunflower oil), marine oils (especially shark liver oil), and synthetic waxes and oils, which have a fatty acid composition similar to that of animal fats and waxes, and preferably to that of human fats and waxes, may also be employed.
In a prefeπed embodiment, the mixture of saturated and unsaturated fatty acids has a composition similar to that of human fat and comprises the following fatty acids: butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, myristoleic acid, palmitic acid, paimitoleic acid, stearic, oleic acid, linoleic acid, linolenic acid, arachidic acid, and gadoleic acid. Preferably, butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, myristoleic acid, palmitic acid, paimitoleic acid, stearic, oleic acid, linoleic acid, linolenic acid, arachidic acid, and gadoleic acid are present in the mixture in about the following percentages by weight, respectively (carbon chain number and number of unsaturations are shown parenthetically, respectively): 0.2%-0.4% (C4), 0.1% (C6), 0.3%-0.8% (Co), 2.2%-3.5% (C10), 0.9%-5.5% (C12), 2.8%-8.5% (C14), 0.1%-0.6% (C14: 1), 23.2%-24.6% (CJ 6), 1.8%-3.0% (C16; 1), 6.9%-9.9% (C] 8), 36.0%-36.5% (Cl g: 1), 20%-20.6% (C18:2), 7.5- 7.8% (C18.3), l.l%-4.9% (C20), and 3.3%-6.4% (C2( ).
In another prefeπed embodiment, the mixture of saturated and unsaturated fatty acids is typically chicken fat comprising the following fatty acids: lauric acid, myristic acid, myristoleic acid, pentadecanoic acid, palmitic acid, paimitoleic acid, margaric acid, margaroleic acid, stearic, oleic acid, linoleic acid, linolenic acid, arachidic acid, and gadoleic acid. Preferably, lauric acid, myristic acid, myristoleic acid, pentadecanoic acid, palmitic acid, paimitoleic acid, margaric acid, margaroleic acid, stearic, oleic acid, linoleic acid, linolenic acid, arachidic acid, and gadoleic acid are present in the mixture in about the following percentages by weight, respectively: 0.1% (C12), 0.8% (C14), 0.2% (C14: ]), 0.1% (C15), 25.3% (C16), 7.2% (C16: 1), 0.1% (C ), 0.1% (C17. j), 6.5% (Cl g), 37.7% (Cl g: 1), 20.6% (C18:2), 0.8% (C18;3), 0.2%
(C20), and 0.3% (C2Q.j), all percentages +/- 10%. In another preferred embodiment, the mixture of saturated and unsaturated fatty acids comprises lecithin. Lecithin (phosphatidylcholine) is a phosphatide found in all living organisms (plants and animals) and is a significant constituent of nervous tissue and brain substance. Lecithin is a mixture of the diglycerides of stearic, palmitic, and oleic acids, linked to the choline ester of phosphoric acid. The product of commerce is predominantly soybean lecithin obtained as a by-product in the manufacturing of soybean oil. Soybean lecithin contains palmitic acid 11.7%, stearic 4.0%, paimitoleic 8.6%, oleic 9.8%, linoleic 55.0%, linolenic 4.0%, C 0 to C2 acids (includes arachidonic) 5.5%. Lecithin may be represented by the foπnula:
CH2OCOR
I CHOCOR
I CH2O-P(O)2-OCH2CH2N+(CH3)3
wherein R is selected from the group consisting of stearic, palmitic, and oleic acid.
The above fatty acids and percentages thereof present in the fatty acid mixture are given as an example. The exact type of fatty acid present in the fatty acid mixture and the exact amount of fatty acid employed in the fatty acid mixture may be varied in order to obtain the result desired in the final product and such variations are now within the capabilities of those skilled in the art without the need for undue experimentation.
The amount of fatty acids present in the therapeutic wound healing compositions of the present invention is a therapeutically effective amount. A therapeutically effective amount of fatty acids is that amount of fatty acids necessary for the inventive composition to prevent and reduce injury to mammalian cells or increase the resuscitation rate of injured mammalian cells. The exact amount of fatty acids employed is subject to such factors as the type and distribution of fatty acids employed in the mixture, the type of condition being treated, and the other ingredients in the composition. In a prefeπed embodiment, the fatty acids are present in the therapeutic wound healing composition in an amount from about 10% to about 50%, preferably from about 20% to about 45%, and more preferably from about 25% to about 40%, by weight of the therapeutic wound healing composition.
In accord with the present invention, the therapeutic wound healing compositions of Embodiment One (I.A-D) for treating mammalian cells may be selected from the group consisting of:
(I.A)(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
(I.B)(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
( C) (a) an antioxidant; and
(b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
(I.D) (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; (b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
Preferably, the wound healing compositions of Embodiment One (I) for treating mammalian cells, preferably epidermal keratinocytes, may be selected from the group consisting of:
(LA) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
(LB) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
( C) (a) an antioxidant; and
(b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
More preferably, the wound healing compositions of Embodiment One
(I) for treating mammalian cells, preferably epidermal keratinocytes, may be selected from the group consisting of: (LA) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
( C) (a) an antioxidant; and
(b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
More preferably, the wound healing compositions of Embodiment One (I) for treating mammalian cells, preferably epidermal keratinocytes, may be selected from the group consisting of:
(LA) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(LB) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. Most preferably, the wourd healing compositions of Embodiment One (I) for treating mammalian cells, preferably epidermal keratinocytes, comprise:
(LA) (a) pyruvate selected from the group consisting of pyruvic acid, phaπnaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
Most preferably, the wound healing compositions of Embodiment One (I) for treating mammalian cells, preferably monocytes, comprise:
(LD) (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
Throughout this disclosure, applicant will suggest various theories or mechanisms by which applicant believes the components in the therapeutic wound healing compositions and the antiviral agent function together in an unexpected synergistic manner to prevent and reduce injury to mammalian cells, increase the resuscitation rate of injured mammalian cells, and reduce viral titers. While applicant may offer various mechanisms to explain the present invention, applicant does not wish to be bound by theory. These theories are suggested to better understand the present invention but are not intended to limit the effective scope of the claims.
In the first aspect of Embodiment One (IA), applicant believes that pyruvate can be transported inside a cell where it can act as an antioxidant to neutralize oxygen radicals in the cell. Pyruvate can also be used inside the cell in the citric acid cycle to provide energy to increase cellular viability, and as a precursor in the synthesis of important biomolecules to promote cellular proliferation. In addition, pyruvate can be used in the multifunction oxidase system to reverse cytotoxicity. Antioxidants, especially lipid-soluble antioxidants, can be absorbed into the cell membrane to neutralize oxygen radicals and thereby protect the membrane. The saturated and unsaturated fatty acids in the present invention are those fatty acids required for the resuscitation of mammalian cells and are readily available for the repair of injured cells and the proliferation of new cells. Cells injured by oxygen radicals need to produce unsaturated fatty acids to repair cellular membranes. However, the production of unsaturated fatty acids by cells requires oxygen. Thus, the injured cell needs high levels of oxygen to produce unsaturated fatty acids and at the same time needs to reduce the level of oxygen within the cell to reduce oxidative injury. By providing the cell with the unsaturated fatty acids needed for repair, the need of the cell for unsaturated fatty acids is reduced and the need for high oxygen levels is also reduced.
The combination of pyruvate inside the cell and an antioxidant in the cellular membrane functions in an unexpected synergistic manner to reduce hydrogen peroxide production in the cell to levels lower than can be achieved by use of either type of component alone. The presence of mixtures of saturated and unsaturated fatty acids in the therapeutic wound healing composition significantly enhances the ability of pyruvate and the antioxidant to inhibit reactive oxygen production. By stabilizing the cellular membrane, unsaturated fatty acids also improve membrane function and enhance pyruvate transport into the cell. Hence, the three components in the therapeutic wound healing composition of the first aspect of Embodiment One (IA) function together in an unexpected synergistic manner to prevent and reduce injury to mammalian cells and increase the resuscitation rate of injured mammalian cells.
In the second aspect of Embodiment One (LB), lactate is employed instead of an antioxidant. Antioxidants react with, and neutralize, oxygen radicals after the radicals are already formed. Lactate, on the other hand, is a component in the cellular feedback mechanism and inhibits the respiratory bursting process to suppress the production of active oxygen species. The combination of pyruvate to neutralize active oxygen species and lactate to suppress the respiratory bursting process functions in a synergistic manner to reduce hydrogen peroxide production in the cell to levels lower than can be achieved by use of either type of component alone. The presence of mixtures of saturated and unsaturated fatty acids in the therapeutic wound healing composition significantly enhances the ability of pyruvate and lactate to inhibit reactive oxygen production. Hence, the three components in the therapeutic wound healing composition in the second aspect of Embodiment One (I.B) function together in a synergistic manner to protect and resuscitate mammalian cells.
In the third aspect of Embodiment One (LC), the presence of mixtures of saturated and unsaturated fatty acids in the therapeutic wound healing composition in this embodiment significantly enhances the ability of the antioxidant to inhibit reactive oxygen production. The combination of an antioxidant to neutralize active oxygen species and fatty acids to rebuild cellular membranes and reduce the need of the cell for oxygen functions in a synergistic manner to reduce hydrogen peroxide production in the cell to levels lower than can be achieved by either type of component alone. Hence, the components in the therapeutic wound healing composition in the third aspect of Embodiment One (LC) function together in a synergistic manner to protect and resuscitate mammalian cells.
In the fourth aspect of Embodiment One (I.D), lactate is employed because the respiratory bursting process is more pronounced in monocytes than in epidermal keratinocytes. The combination of lactate to suppress the respiratory bursting process and an antioxidant to neutralize active oxygen species functions in a synergistic manner to reduce hydrogen peroxide production in the cell to levels lower than can be achieved by either component alone. The presence of mixtures of saturated and unsaturated fatty acids in the therapeutic wound healing composition in this embodiment significantly enhances the ability of lactate and the antioxidant to inhibit reactive oxygen production. Hence, the three components in the therapeutic wound healing composition in the fourth aspect of Embodiment One (I.D) function together in an unexpected synergistic manner to protect and resuscitate mammalian cells. Accordingly, the combination of ingredients set out in the above embodiments functions together in an enhanced manner to prevent and reduce injury to mammalian cells and increase the resuscitation rate of injured mammalian cells. The therapeutic effect of the combination of the components in each of the above embodiments is markedly greater than that expected by the mere addition of the individual therapeutic components. Hence, applicant's therapeutic wound healing compositions for treating mammalian cells have the ability to decrease intracellular levels of hydrogen peroxide production, increase cellular resistance to cytotoxic agents, increase rates of cellular proliferation, and increase cellular viability.
B. Methods For Making
The Therapeutic Wound Healing Compositions
Of Embodiment One (LA-D)
The present invention extends to methods for making the therapeutic wound healing compositions of Embodiment One (IA-D). In general, a therapeutic wound healing composition is made by forming an admixture of the components of the composition. In a first aspect of Embodiment One (IA), a therapeutic wound healing composition is made by forming an admixture of (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. In a second aspect of Embodiment One (I.B), a therapeutic wound healing composition is made by forming an admixture of (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. In a third aspect of Embodiment
One (LC), a therapeutic wound healing composition is made by forming an admixture of (a) an antioxidant and (b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells. In a fourth aspect of Embodiment One (I.D), a therapeutic wound healing composition is made by forming an admixture of (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
For some applications, the admixture may be formed in a solvent such as water, and a surfactant may be added if required. If necessary, the pH of the solvent is adjusted to a range from about 3.5 to about 8.0, and preferably from about 4.5 to about 7.5, and more preferably about 6.0 to about 7.4. The admixture is then sterile filtered. Other ingredients may also be incorporated into the therapeutic wound healing composition as dictated by the nature of the desired composition as well known by those having ordinary skill in the art. The ultimate therapeutic wound healing compositions are readily prepared using methods generally known in the pharmaceutical arts.
In a prefeπed embodiment, the invention is directed to a method for preparing a therapeutic wound healing composition (IA) for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of admixing the following ingredients:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for die resuscitation of injured mammalian cells. C. Methods For Employing
The Therapeutic Wound Healing Compositions
Of Embodiment One (LA-D)
The present invention extends to methods for employing the therapeutic wound healing compositions of Embodiment One (I) in vivo and in vitro. In general, a therapeutic wound healing composition is employed by contacting the therapeutic composition with mammalian cells.
In a first aspect of Embodiment One (I. A), the invention is directed to a method for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of (A) providing a therapeutic wound healing composition which comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells, and (B) contacting the therapeutic wound healing composition with the mammalian cells.
In a second aspect of Embodiment One (LB), the invention is directed to a method for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of (A) providing a therapeutic wound healing composition which comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells, and (B) contacting the therapeutic wound healing composition with the mammalian cells.
In a third aspect of Embodiment One (LC), the invention is directed to a method for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of (A) providing a therapeutic wound healing composition which comprises (a) an antioxidant, and (b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells, and (B) contacting the therapeutic wound healing composition with the mammalian cells.
In a fourth aspect of Embodiment One (I.D), the invention is directed to a method for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of (A) providing a therapeutic wound healing composition which comprises (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells, and (B) contacting die therapeutic wound healing composition with the mammalian cells.
In a preferred embodiment, the invention is directed to a method for healing a wound in a mammal which comprises the steps of:
(A) providing a therapeutic wound healing composition (LA) which comprises: (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells; and
(B) contacting the therapeutic wound healing composition with the wound.
The types of wounds which may be healed using the wound healing compositions of Embodiment One (LA-D) of the present invention are those which result from an injury which causes epidermal damage such as incisions, wounds in which the skin is broken by a cutting instrument, and lacerations, wounds in which the skin is broken by a dull or blunt instrument. The therapeutic compositions may also be used to treat various dermatological disorders such as hyperkeratosis, photo-aging, bums, donor site wounds from skin transplants, ulcers (cutaneous, decubitus, venous stasis, and diabetic), psoriasis, skin rashes, and sunburn photoreactive processes. The topical therapeutic compositions may also be used orally in the form of a mouth wash or spray to protect and accelerate the healing of injured oral tissue such as mouth sores and bums. The topical therapeutic compositions may further be used in ophthalmological preparations to treat wounds such as those which result from comeal ulcers, radialkeratotomy, comeal transplants, epikeratophakia and other surgically induced wounds in the eye. The topical therapeutic compositions may in addition be used in anorectal creams and suppositories to treat such conditions as pruritus and, proctitis, anal fissures, and hemorrhoids. In a prefeπed embodiment, die therapeutic compositions are used to treat wounds such as incisions and lacerations.
The wound healing compositions of Embodiment One (LA-D) of the present invention may be utilized in topical products, ingestible products, and tissue culture medium to protect mammalian cells and increase the resuscitation rate of injured mammalian cells. For example, the therapeutic wound healing compositions may be used in topical skin care products to protect and increase the resuscitation rate of skin tissue such as in the treatment of various dermatological disorders such as hyperkeratosis, photo-aging, and sunburn photoreactive processes. Injuτy to skin can occur for a variety of reasons. Injury often occurs to individuals who wash their hands often, to individuals who are exposed to stressful environmental conditions (overexposure to sun or chemicals), or to the elderly or individuals with an underlining disease. The addition of die wound healing compositions of the present invention to a lotion provides a source of antioxidants to the skin which would protect die skin from the harmful effects of UV light, chemicals, and severe drying. The wound healing compositions can be used for the following indications: a) Moisturizing and protecting; b) Healing dry cracked skin; c) Treating irritated skin such as diaper rash; d) Healing severe dry skin due to other diseases (venous dermatitis); e) Treating psoriasis and other hyperproliferative diseases; f) Protecting skin from UV light damage (antioxidant skin replacement); g) Treating seboπheic conditions; and h) Treating shaving wounds in an after shave lotion. The topical therapeutic wound healing compositions may also be used orally in the form of a mouth wash or spray to protect and accelerate the healing of injured oral tissue such as mouth sores and bu s. The topical therapeutic wound healing compositions may further be used in ophthalmological preparations such as eye care products to neutralize hydrogen peroxide used in the cleaning of contact lenses.
The topical therapeutic wound healing compositions may in addition be used in anorectal creams and suppositories to treat such conditions as pruritus and, proctitis, anal fissures, and hemoπhoids. Initially as white blood cells enter a wound site, the cells release oxygen radicals, depleting die antioxidants at die wound site, thus impairing the healing process. Incorporating the wound healing compositions of die present invention into a wound healing formulation would facilitate healing by providing die site with usable antioxidants, and a source of fatty acids needed for membrane repair. The wound healing compositions can be used for the following indications: a) Healing of cuts and scrapes; b) Bu s (heals bums with less scaring and scabbing); c) Decubitus ulcers; d) Bed sores, pressure ulcers; e) Fissures, Hemoπhoids; f) Use in combination with immunostimulators (simulated healing in healing deficient people); g) Post surgical wounds; h) Bandages; i) Diabetic ulcers; j) Venous ulceration; and k) Use in combination with wound cleansing agents.
The therapeutic wound healing compositions may also be used in ingestible products to protect and increase the resuscitation rate of erosions, stomach ulcers, and hemoπhages in the gastric mucosa. Other ingestible therapeutic products include: stroke medications; autoimmune disease medications; arthritis medications; ulcer medications; cancer medications (cytotoxic agents); heart medication to improve regional ventricular function and restore normal heart rate and pressure functions; lung medication to repair injured tissue; liver medication to suppress lipogenesis of alcoholic origin and prevent hepatic steatosis; kidney medication to suppress urinary calculi (kidney stones); detoxification medication to antagonize heavy metal poisoning, cyanide poisoning, sodium sulfide poisoning, other types of poisoning. ; and reduce and neutralize the production of oxygen radicals which produces injury to tissue, to protect and further enhance the resuscitation rate of the injured mammalian cells. The therapeutic wound healing compositions may be used in ingestible products to treat inflammatory diseases such as hepatitis, gastritis, colitis, esophagitis, arthritis, and pancreatitis.
The therapeutic wound healing compositions of the present invention may also be used in tissue culture media and organ transplant media to prevent and reduce injury to mammalian cells and increase the resuscitation rate of injured mammalian cells. Tissue cultures and transplant organs encounter reactive oxygen species generated in the culture media by die injured cells. Organs particularly susceptible to oxidative damage during transport and transplantation due to reperfusion injury following ischemia are corneas, livers, hearts, and kidneys. The tiierapeutic wound healing compositions may be useful to abrogate reperfusion injury to such transplant organs.
In a specific embodiment, die invention is directed to a method for preserving mammalian cells in a culture medium which comprises the steps of:
(A) providing a tiierapeutic wound healing composition selected from the group of consisting of:
(IA) (a) pyruvate selected from die group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures tiiereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
(LB) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; and (c) a mixture of saturated and unsaturated fatty acids wherein die fatty acids are tiiose fatty acids required for die repair of cellular membranes and resuscitation of mammalian cells; (LC) (a) an antioxidant; and
(b) a mixture of saturated and unsaturated fatty acids wherein die fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
(I.D) (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are tiiose fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein die fatty acids are tiiose fatty acids required for the resuscitation of injured mammalian cells;
(B) providing mammalian cells in a culture medium; and
(C) contacting the therapeutic wound healing composition from step (A) witii die mammalian cells in the culture medium from step (B).
D. Formulations Of
The Therapeutic Wound Healing Compositions Of Embodiment One (LA-D)
Once prepared, die inventive tiierapeutic wound healing compositions of Embodiment One (LA-D) may be stored for future use or may be formulated in effective amounts with pharmaceutically acceptable carriers to prepare a wide variety of pharmaceutical compositions. Examples of pharmaceutically acceptable carriers are pharmaceutical appliances, topical vehicles (non-oral and oral), and ingestible vehicles.
Examples of pharmaceutical appliances are sutures, staples, gauze, bandages, bum dressings, artificial skins, liposome or micell formulations, microcapsules, aqueous vehicles for soaking gauze dressings, and the like, and mixtures thereof. Non-oral topical compositions employ non-oral topical vehicles, such as creams, gels formulations, foams, ointments and sprays, salves, and films, which are intended to be applied to die skin or body cavity and are not intended to be taken by mouth. Oral topical compositions employ oral vehicles, such as mouthwashes, rinses, oral sprays, suspensions, and dental gels, which are intended to be taken by mouth but are not intended to be ingested. Ingestible compositions employ ingestible or partly ingestible vehicles such as confectionery bulking agents which include hard and soft confectionery such as lozenges, tablets, toffees, nougats, suspensions, chewy candies, and chewing gums.
In one form of the invention, die therapeutic wound healing composition is incorporated into a pharmaceutical appliance which may be in the form of sutures, staples, gauze, bandages, bum dressings, artificial skins, liposome or micell formulations, microcapsules, aqueous vehicles for soaking gauze dressings, and the like, and mixtures thereof. A variety of traditional ingredients may optionally be included in die pharmaceutical composition in effective amounts such as buffers, preservatives, tonicity adjusting agents, antioxidants, polymers for adjusting viscosity or for use as extenders, and excipients, and die like. Specific illustrative examples of such traditional ingredients include acetate and borate buffers; thimerosal, sorbic acid, methyl and propyl paraben and chlorobutanol preservatives; sodium chloride and sugars to adjust die tonicity; and excipients such as mannitol, lactose and sucrose. Other conventional pharmaceutical additives known to those having ordinary skill in the pharmaceutical arts may also be used in the pharmaceutical composition.
In accordance with this invention, tiierapeutically effective amounts of the therapeutic wound healing compositions of die present invention may be employed in die pharmaceutical appliance. These amounts are readily determined by tiiose skilled in the art widiout the need for undue experimentation. The exact amount of the therapeutic wound healing composition employed is subject to such factors as the type and concentration of die therapeutic wound healing composition and d e type of pharmaceutical appliance employed. Thus, the amount of tiierapeμtic wound healing composition may be varied in order to obtain the result desired in the final product and such variations are within the capabilities of tiiose skilled in the art without die need for undue experimentation. In a prefeπed embodiment, die pharmaceutical composition will comprise die tiierapeutic wound healing composition in an amount from about 0.1% to about 5%, by weight of the pharmaceutical composition. In a more preferred embodiment, the pharmaceutical composition will comprise die tiierapeutic wound healing composition in an amount from about 0.1% to about 3%, by weight of the pharmaceutical composition. In a most preferred embodiment, the pharmaceutical composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 1%, by weight of the pharmaceutical composition.
The present invention extends to methods for making the pharmaceutical compositions. In general, a pharmaceutical composition is made by contacting a therapeutically effective amount of a tiierapeutic wound healing composition with a pharmaceutical appliance and the other ingredients of die final desired pharmaceutical composition. The therapeutic wound healing composition may be in a solvent and may be absorbed onto a pharmaceutical appliance.
Other ingredients will usually be incorporated into the composition as dictated by the nature of the desired composition as well known by those having ordinary skill in the art. The ultimate pharmaceutical compositions are readily prepared using methods generally known in the pharmaceutical arts.
In another form of the invention, die therapeutic wound healing composition is incorporated into a non-oral topical vehicle which may be in the form of a cream, gel, foam, ointment, spray, and die like. Typical non-toxic non-oral topical vehicles known in the pharmaceutical arts may be used in die present invention. The prefeπed non-oral topical vehicles are water and pharmaceutically acceptable water- miscible organic solvents such as ethyl alcohol, isopropyl alcohol, propylene glycol, glycerin, and the like, and mixtures of these solvents. Water-alcohol mixtures are particularly preferred and are generally employed in a weight ratio from about 1 : 1 to about 20:1, preferably from about 3:1 to about 20:1, and most preferably from about 3:1 to about 10:1, respectively. The non-oral topical therapeutic wound healing compositions may also contain conventional additives employed in tiiose products. Conventional additives include humectants, emollients, lubricants, stabilizers, dyes, and perfumes, providing the additives do not interfere with the therapeutic properties of the tiierapeutic wound healing composition.
Suitable humectants useful in the non-oral topical therapeutic wound healing compositions include glycerin, propylene glycol, polyethylene glycol, sorbitan, fructose, and die like, and mixtures thereof. Humectants, when employed, may be present in amounts from about 10% to about 20%, by weight of the topical therapeutic wound healing composition.
The coloring agents (colors, colorants) useful in the non-oral topical therapeutic wound healing composition are used in amounts effective to produce die desired color. These coloring agents include pigments which may be incorporated in amounts up to about 6% by weight of the non-oral topical therapeutic wound healing composition. A prefeπed pigment, titanium dioxide, may be incorporated in amounts up to about 2%, and preferably less than about 1 %, by weight of the non-oral topical therapeutic wound healing composition. The coloring agents may also include natural food colors and dyes suitable for food, drug and cosmetic applications. These coloring agents are known as F.D.& C. dyes and lakes. The materials acceptable for the foregoing uses are preferably water-soluble. Illustrative nonlimiting examples include die indigoid dye known as F.D.& C. Blue No.2, which is the disodium salt of 5,5-indigotindisulfonic acid. Similarly, the dye known as F.D.& C. Green No.l comprises a triphenylmethane dye and is die monosodium salt of 4-[4-(N-ethyl-ι>- sulfoniumbenzylamino) diphenylmethylene]-[ 1 -(N-ethyl-N-g-sulfoniumbenzyl)-delta- 2,5-cyclohexadieneimine]. A full recitation of all F.D.& C. coloring agents and dieir coπesponding chemical structures may be found in the Kirk-Othmer Encyclopedia of Chemical Technology, 3rd Edition, in volume 5 at pages 857-884, which text is incorporated herein by reference. In accordance witii this invention, tiierapeutically effective amounts of the therapeutic wound healing compositions of die present invention may be admixed witii a non-oral topical vehicle to form a topical therapeutic wound healing composition. These amounts are readily determined by tiiose skilled in the art without the need for undue experimentation. In a preferred embodiment, die non-oral topical therapeutic wound healing compositions will comprise die therapeutic wound healing composition in an amount from about 0.1% to about 10% and a non-oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the non-oral topical therapeutic wound healing composition. In a more preferred embodiment, the non-oral topical therapeutic wound healing compositions will comprise die therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most preferred embodiment, the non-oral topical therapeutic wound healing compositions will comprise the tiierapeutic wound healing composition in an amount from about 0.1% to about 2%, and a non-oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the non-oral topical tiierapeutic wound healing composition.
The present invention extends to metiiods for preparing the non-oral topical therapeutic wound healing compositions. In such a method, die non-oral topical tiierapeutic wound healing composition is prepared by admixing a tiierapeutically effective amount of the therapeutic wound healing composition of die present invention and a non-oral topical vehicle. The final compositions are readily prepared using standard methods and apparatus generally known by those skilled in die pharmaceutical arts. The apparatus useful in accordance witii the present invention comprises mixing apparatus well known in the pharmaceutical arts, and therefore the selection of the specific apparatus will be apparent to the artisan.
In another form of the invention, the therapeutic wound healing composition is incorporated into an oral topical vehicle which may be in the form of a mouthwash, rinse, oral spray, suspension, dental gel, and die like. Typical non-toxic oral vehicles known in the pharmaceutical arts may be used in die present invention.
The preferred oral vehicles are water, etiianol, and water-etiianol mixtures. The water- ethanol mixtures are generally employed in a weight ratio from about 1:1 to about 20:1, preferably from about 3:1 to about 20:1, and most preferably from about 3:1 to about 10:1, respectively. The pH value of the oral vehicle is generally from about 4 to about 7, and preferably from about 5 to about 6.5. An oral topical vehicle having a pH value below about 4 is generally irritating to the oral cavity and an oral vehicle having a pH value greater than about 7 generally results in an unpleasant mouth feel.
The oral topical therapeutic wound healing compositions may also contain conventional additives normally employed in tiiose products. Conventional additives include a fluorine providing compound, a sweetening agent, a flavoring agent, a coloring agent, a humectant, a buffer, and an emulsifier, providing the additives do not interfere with the therapeutic properties of the therapeutic wound healing composition.
The coloring agents and humectants, and die amounts of these additives to be employed, set out above as useful in the non-oral topical therapeutic wound healing composition may be used in the oral topical therapeutic wound healing composition.
Fluorine providing compounds may be fully or slightly water soluble and are characterized by their ability to release fluoride ions or fluoride containing ions in water and by tiieir lack of reaction with other components in the composition. Typical fluorine providing compounds are inorganic fluoride salts such as water-soluble alkali metal, alkaline earth metal, and heavy metal salts, for example, sodium fluoride, potassium fluoride, ammonium fluoride, cuprous fluoride, zinc fluoride, stannic fluoride, stannous fluoride, barium fluoride, sodium fluorosilicate, ammonium fluorosilicate, sodium fluorozirconate, sodium monofluorophosphate, aluminum mono- and di-fluorophosphates and fluorinated sodium calcium pyrophosphate. Alkali metal fluorides, tin fluoride and monofluorophosphates, such as sodium and stannous fluoride, sodium monofluorophosphate and mixtures thereof, are prefeπed. The amount of fluorine providing compound present in d e present oral topical therapeutic wound healing composition is dependent upon the type of fluorine providing compound employed, die solubility of the fluorine compound, and die nature of the final oral therapeutic wound healing composition. The amount of fluorine providing compound used must be a nontoxic amount. In general, the fluorine providing compound when used will be present in an amount up to about 1%, preferably from about 0.001% to about 0.1%, and most preferably from about 0.001% to about 0.05%, by weight of the oral topical therapeutic wound healing composition.
When sweetening agents (sweeteners) are used, tiiose sweeteners well known in the art, including both natural and artificial sweeteners, may be employed. The sweetening agent used may be selected from a wide range of materials including water-soluble sweetening agents, water-soluble artificial sweetening agents, water- soluble sweetening agents derived from naturally occurring water-soluble sweetening agents, dipeptide based sweetening agents, and protein based sweetening agents, including mixtures thereof. Without being limited to particular sweetening agents, representative categories and examples include:
(a) water-soluble sweetening agents such as monosaccharides, disaccharides and polysaccharides such as xylose, ribose, glucose (dextrose), mannose, galactose, fructose (levulose), sucrose (sugar), maltose, invert sugar (a mixture of fructose and glucose derived from sucrose), partially hydrolyzed starch, co syrup solids, dihydrochalcones, monellin, steviosides, and glycyπhizin, and mixtures thereof;
(b) water-soluble artificial sweeteners such as soluble saccharin salts, i.e., sodium or calcium saccharin salts, cyclamate salts, the sodium, ammonium or calcium salt of 3 ,4-dihydro-6-methyl- 1 ,2,3-oxathiazine-4-one-2,2-dioxide, die potassium salt of 3 ,4-dihydro-6-medιyl- 1 ,2 ,3-oxathiazine-4-one-2,2-dioxide ( Acesulfame-K), the free acid form of saccharin, and the like;
(c) dipeptide based sweeteners, such as L-aspartic acid derived sweeteners, such as L-aspartyl-L-phenylalanine methyl ester (Aspartame) and materials described in United States Patent No. 3,492,131 , L-Alpha-aspaτtyl-N-(2,2,4,4- tetramethyl-3-thietanyl)-D-alanin-amide hydrate (Alitame), methyl esters of L-aspartyl- L-phenylglycerineandL-aspartyl-L-2,5-dihydrophenyl-glycine, L-aspartyl-2,5-dihydro- L-phenylalanine; L-aspartyl-L-(l-cyclohexen)-alanine, and die like;
(d) water-soluble sweeteners derived from naturally occurring water- soluble sweeteners, such as chlorinated derivatives of ordinary sugar (sucrose), e.g., chlorodeoxysugar derivatives such as derivatives of chlorodeoxysucrose or chlorodeoxygalactosucrose, known, for example, under the product designation of Sucralose; examples of chlorodeoxysucrose and chlorodeoxygalacto-sucrose derivatives include but are not limited to: l-chloro-l'-deoxysucrose; 4-chloro-4-deoxy-Alpha-D- galacto-pyranosyl-Alpha-D-fructofuranoside, or 4-chloro-4-deoxygalactosucrose; 4- chloro-4-deoxy-Alpha-D-galacto-pyranosyl- 1 -chloro- 1 -deoxy-B-D-fructo-furanoside, or
4, 1 ' -dichloro-4, 1 ' -dideoxygalactosucrose; 1 ' ,6' -dichloro- 1 ' ,6' -dideoxysucrose; 4-chloro- 4-deoxy-Alpha-D-galacto-pyranosyl-l ,6-dichloro-l ,6-dideoxy-β-D-fructo-furanoside, or 4,1 ',6'-trichloro-4, 1 ',6'-trideoxygalacto-sucrose; 4,6-dichloro-4,6-dideoxy-Alpha-D- galacto-pyranosyl-6-chloro-6-deoxy-β-D-fructofuranoside, or 4,6,6'-trichloro-4,6,6'- trideoxygalactosucrose; 6, ,6'-trichloro-6, ,6'-trideoxysucrose; 4,6-dichloro-4,6- dideoxy-Alpha-D-galacto-pyranosyl-1 ,6-dichloro-l ,6-di-deoxy-β-D-fructofuranoside, or 4,6, 1 ',6'-tetrachloro-4,6, 1 ',6'-tetradeoxygalacto-sucrose; and 4,6, 1 ',6'-tetrachloro- 4,6, 1 ',6'-tetradeoxy-sucrose; and
(e) protein based sweeteners such as thaumaoccous danielli (Thaumatin I and II).
In general, an effective amount of sweetening agent is utilized to provide the level of sweetness desired in die particular oral topical therapeutic wound healing composition, and this amount will vary with die sweetener selected and die final oral therapeutic product desired. The amount of sweetener normally present is in the range from about 0.0025% to about 90%, by weight of the oral topical tiierapeutic wound healing composition, depending upon the sweetener used. The exact range of amounts for each type of sweetener is well known in the art and is not the subject of the present invention.
The flavoring agents (flavors, flavorants) which may be used include tiiose flavors known to the skilled artisan, such as natural and artificial flavors. Suitable flavoring agents include mints, such as peppermint, cirrus flavors such as orange and lemon, artificial vanilla, cinnamon, various fruit flavors, both individual and mixed, and die like.
The amount of flavoring agent employed in the oral topical therapeutic wound healing composition is normally a matter of preference subject to such factors as the type of final oral therapeutic wound healing composition, the individual flavor employed, and die strength of flavor desired. Thus, the amount of flavoring may be varied in order to obtain the result desired in the final product and such variations are within the capabilities of those skilled in the art without the need for undue experimentation. The flavoring agents, when used, are generally utilized in amounts tiiat may, for example, range in amounts from about 0.05% to about 6%, by weight of the oral topical therapeutic wound healing composition.
Suitable buffer solutions useful in die non-oral topical tiierapeutic wound healing compositions include citric acid-sodium citrate solution, phosphoric acid- sodium phosphate solution, and acetic acid-sodium acetate solution in amounts up to about 1%, and preferably from about 0.05% to about 0.5% by weight of the oral topical therapeutic wound healing composition.
In accordance witii this invention, therapeutically effective amounts of the therapeutic wound healing compositions of the present invention may be admixed witii an oral topical vehicle to form a topical therapeutic wound healing composition. These amounts are readily determined by tiiose skilled in the art without the need for undue experimentation. In a prefeπed embodiment, die oral topical therapeutic wound healing compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 10% and a oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the oral topical therapeutic wound healing composition. In a more prefeπed embodiment, the oral topical therapeutic wound healing compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most preferred embodiment, the oral topical therapeutic wound healing compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and a oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the oral topical tiierapeutic wound healing composition.
The present invention extends to methods for preparing the oral topical therapeutic wound healing compositions. In such a method, die oral topical therapeutic wound healing composition is prepared by admixing a therapeutically effective amount of the therapeutic wound healing composition of the present invention and an oral topical vehicle. The final compositions are readily prepared using standard methods and apparatus generally known by those skilled in the pharmaceutical arts. The apparatus useful in accordance with the present invention comprises mixing apparatus well known in the pharmaceutical arts, and tiierefore the selection of the specific apparatus will be apparent to the artisan.
In a prefeπed embodiment, an oral topical therapeutic wound healing composition is made by first dissolving coloring agents, sweetening agents, and similar additives in water. The therapeutic wound healing composition is then admixed witii the aqueous solution. Then sufficient water or ethanol, or mixtures of water and ethanol, are added to the solution with mixing until the final solution volume is reached. In a more prefeπed embodiment, the therapeutic wound healing composition is added to die solution as the final ingredient. The final oral topical therapeutic wound healing compositions are readily prepared using metiiods generally known in the pharmaceutical arts.
The oral therapeutic wound healing composition may also be in the form of dental gel. As used herein, the term "gel" means a solid or semisolid colloid which contains considerable quantities of water. The colloid particles in a gel are linked together in a coherent meshwork which immobilizes the water contained inside d e meshwork. The dental gel compositions of the present invention may contain the conventional additives set out above for oral topical therapeutic wound healing compositions such as mouthwashes, rinses, oral sprays, and suspensions and, in addition, may contain additional additives such as a polishing agent, a desensitizing agent, and die like, providing die additional additives do not interfere with die tiierapeutic properties of the therapeutic wound healing composition.
In a dental gel composition, the oral vehicle generally comprises water, typically in an amount from about 10% to about 90%, by weight of the dental gel composition. Polyethylene glycol, propylene glycol, glycerin, and mixtures thereof may also be present in the vehicle as humectants or binders in amounts from about 18% to about 30%, by weight of the dental gel composition. Particularly preferred oral vehicles comprise mixtures of water with polyethylene glycol or water with glycerin and polypropylene glycol.
The dental gels of die present invention include a gelling agent (diickening agent) such as a natural or synthetic gum or gelatin. Gelling agents such as hydroxyethyl cellulose, methyl cellulose, glycerin, carboxypolymethylene, and gelatin and die like, and mixtures thereof may be used. The preferred gelling agent is hydroxyediyl cellulose. Gelling agents may be used in amounts from about 0.5% to about 5%, and preferably from about 0.5% to about 2%, by weight of the dental gel composition.
The dental gel compositions of the present invention may also include a polishing agent. In clear gels, a polishing agent of colloidal silica and/or alkali metal aluminosilicate complexes is prefeπed since these materials have refractive indices close to the refractive indices of the gelling systems commonly used in dental gels. In non-clear gels, a polishing agent of calcium carbonate or calcium dihydrate may be used. These polishing agents may be used in amounts up to about 75%, and preferably in amounts up to about 50%, by weight of the dental gel composition. The dental gel may also contain a desensitizing agent such as a combination of citric acid and sodium citrate. Citric acid may be used in an amount from about 0.1% to about 3%, and preferably from about 0.2% to about 1%, by weight, and sodium citrate may be used in an amount from about 0.3% to about 9%, and preferably from about 0.6% to about 3%, by weight of the dental gel composition.
In accordance with this invention, therapeutically effective amounts of the therapeutic wound healing compositions of the present invention may be admixed into the dental gel compositions. These amounts are readily determined by tiiose skilled in the art without the need for undue experimentation. In a prefeπed embodiment, die dental gel compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 10% and an oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the dental gel composition. In a more preferred embodiment, die dental gel compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most prefeπed embodiment, the dental gel compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and an oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the dental gel composition.
The present invention extends to methods for preparing the therapeutic dental gel compositions. In such a method, the dental gel composition is prepared by admixing a tiierapeutically effective amount of the therapeutic wound healing composition of die present invention and an oral topical vehicle. The final compositions are readily prepared using methods generally known by those skilled in the dental and pharmaceutical arts. The apparatus useful in accordance witii the present invention comprises mixing apparatus well known in the pharmaceutical arts, and therefore the selection of the specific apparatus will be apparent to the artisan.
In a prefeπed embodiment, a therapeutic dental gel composition is made by first dispersing a gelling agent in a humectant or water, or a mixture of both, then admixing to the dispersion an aqueous solution of the water-soluble additives such as the fluorine providing compound, sweeteners and die like, then adding die polishing agent, and lastly admixing the flavoring agent and the therapeutic wound healing composition. The final gel mixture is then tubed or otherwise packaged. The liquids and solids in a gel product are proportioned to form a creamy or gelled mass which is extrudable from a pressurized container or from a collapsible tube. The final therapeutic wound healing compositions are readily prepared using metiiods generally known in the pharmaceutical arts.
In yet another form of the invention, the therapeutic wound healing composition is incorporated into an ingestible vehicle. The ingestible vehicle may be a confectionery bulking agent in the form of lozenges, tablets, toffees, nougats, suspensions, chewy candies, chewing gums, and die like. The pharmaceutically acceptable carriers may be prepared from a wide range of materials including, but not limited to, diluents, binders and adhesives, lubricants, disintegrants, coloring agents, bulking agents, flavoring agents, sweetening agents and miscellaneous materials such as buffers and adsorbents tiiat may be needed in order to prepare a particular therapeutic confection.
The preparation of confectionery formulations is historically well known and has changed little through the years. Confectionery items have been classified as eitiier "hard" confectionery or "soft" confectionery. The therapeutic wound healing compositions of the present invention can be incorporated into confectionery compositions by admixing the inventive composition into conventional hard and soft confections.
As used herein, die term confectionery material means a product containing a bulking agent selected from a wide variety of materials such as sugar, com syrup, and in the case of sugarless bulking agents, sugar alcohols such as sorbitol and mannitol and mixtures thereof. Confectionery material may include such exemplary substances as lozenges, tablets, toffee, nougat, suspensions, chewy candy, chewing gum and the like. The bulking agent is present in a quantity sufficient to bring the total amount of composition to 100%. In general, the bulking agent will be present in amounts up to about 99.98%, preferably in amounts up to about 99.9%, and more preferably in amounts up to about 99%, by weight of the ingestible therapeutic wound healing composition.
Lozenges are flavored medicated dosage forms intended to be sucked and held in the mouth. Lozenges may be in the form of various shapes such as flat, circular, octagonal and biconvex forms. The lozenge bases are generally in two forms: hard boiled candy lozenges and compressed tablet lozenges.
Hard boiled candy lozenges may be processed and formulated by conventional means. In general, a hard boiled candy lozenge has a base composed of a mixture of sugar and other carbohydrate bulking agents kept in an amorphous or glassy condition. This amorphous or glassy form is considered a solid syrup of sugars generally having from about 0.5% to about 1.5% moisture. Such materials normally contain up to about 92% com syrup, up to about 55% sugar and from about 0.1% to about 5% water, by weight of the final composition. The syrup component is generally prepared from com syrups high in fructose, but may include other materials. Further ingredients such as flavoring agents, sweetening agents, acidulants, coloring agents and die like may also be added.
Boiled candy lozenges may also be prepared from non-fermentable sugars such as sorbitol, mannitol, and hydrogenated com syrup. Typical hydrogenated co syrups are Lycasin, a commercially available product manufactured by Roquette Corporation, and Hystar, a commercially available product manufactured by Lonza, Inc.
The candy lozenges may contain up to about 95% sorbitol, a mixture of sorbitol and mannitol in a ratio from about 9.5:0.5 up to about 7.5:2.5, and hydrogenated com syrup up to about 55%, by weight of the solid syrup component.
Boiled candy lozenges may be routinely prepared by conventional methods such as those involving fire cookers, vacuum cookers, and scraped-surface cookers also refeπed to as high speed atmospheric cookers. Fire cookers involve the traditional method of making a boiled candy lozenge base. In this method, the desired quantity of carbohydrate bulking agent is dissolved in water by heating d e agent in a kettle until the bulking agent dissolves. Additional bulking agent may then be added and cooking continued until a final temperature of 145°C. to 156°C. is achieved. The batch is then cooled and worked as a plastic-like mass to incorporate additives such as flavors, colorants and die like.
A high-speed atmospheric cooker uses a heat-exchanger surface which involves spreading a film of candy on a heat exchange surface, the candy is heated to
165°C. to 170°C. in a few minutes. The candy is tiien rapidly cooled to 100°C. to 120°C. and worked as a plastic-like mass enabling incorporation of the additives, such as flavors, colorants and die like.
In vacuum cookers, the carbohydrate bulking agent is boiled to 125°C. to 132°C, vacuum is applied and additional water is boiled off without extra heating. When cooking is complete, the mass is a semi-solid and has a plastic-like consistency.
At this point, flavors, colorants, and other additives are admixed in the mass by routine mechanical mixing operations.
The optimum mixing required to uniformly mix the flavoring agents, coloring agents and other additives during conventional manufacturing of boiled candy lozenges is determined by the time needed to obtain a uniform distribution of the materials. Normally, mixing times of from 4 to 10 minutes have been found to be acceptable.
Once the boiled candy lozenge has been properly tempered, it may be cut into workable portions or formed into desired shapes. A variety of forming techniques may be utilized depending upon the shape and size of the final product desired. A general discussion of the composition and preparation of hard confections may be found in HA. Lieberman, Pharmaceutical Dosage Forms: Tablets, Volume I (1 80), Marcel Dekker, Inc., New York, N.Y. at pages 339 to 469, which disclosure is incorporated herein by reference. The apparatus useful in accordance witii the present invention comprises cooking and mixing apparatus well known in the confectionery manufacturing arts, and dierefore the selection of the specific apparatus will be apparent to the artisan.
In contrast, compressed tablet confections contain particulate materials and are formed into structures under pressure. These confections generally contain sugars in amounts up to about 95%, by weight of the composition, and typical tablet excipients such as binders and lubricants as well as flavoring agents, coloring agents and the like.
In addition to hard confectionery materials, the lozenges of the present invention may be made of soft confectionery materials such as those contained in nougat. The preparation of soft confections, such as nougat, involves conventional methods, such as the combination of two primary components, namely (1) a high boiling syrup such as a co syrup, hydrogenated starch hydrolysate or the like, and (2) a relatively light textured frappe, generally prepared from egg albumin, gelatin, vegetable proteins, such as soy derived compounds, sugarless milk derived compounds such as milk proteins, and mixtures thereof. The frappe is generally relatively light, and may, for example, range in density from about 0.5 to about 0.7 grams/cc.
The high boiling syrup, or "bob syrup" of the soft confectionery is relatively viscous and has a higher density than the frappe component, and frequently contains a substantial amount of carbohydrate bulking agent such as a hydrogenated starch hydrolysate. Conventionally, the final nougat composition is prepared by the addition of the "bob syrup" to the frappe under agitation, to form the basic nougat mixture. Further ingredients such as flavoring agents, additional carbohydrate bulking agent, coloring agents, preservatives, medicaments, mixtures thereof and the like may be added thereafter also under agitation. A general discussion of the composition and preparation of nougat confections may be found in B.W. Minifie, Chocolate, Cocoa and Confectionery: Science and Technology. 2nd edition, AVI Publishing Co., Inc.,
Westport, Conn. (1980), at pages 424-425, which disclosure is incorporated herein by reference. The procedure for preparing the soft confectionery involves known procedures. In general, the frappe component is prepared first and thereafter the syrup component is slowly added under agitation at a temperature of at least about 65°C, and preferably at least about 100°C. The mixture of components is continued to be mixed to form a uniform mixture, after which the mixture is cooled to a temperature below
80°C, at which point, the flavoring agent may be added. The mixture is further mixed for an additional period until it is ready to be removed and formed into suitable confectionery shapes.
The ingestible therapeutic wound healing compositions may also be in the form of a pharmaceutical suspension. Pharmaceutical suspensions of this invention may be prepared by conventional methods long established in die art of pharmaceutical compounding. Suspensions may contain adjunct materials employed in formulating the suspensions of the art. The suspensions of the present invention can comprise: (a) preservatives such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), benzoic acid, ascorbic acid, metiiyl paraben, propyl paraben, tocopherols, and die like, and mixtures thereof. Preservatives are generally present in amounts up to about 1%, and preferably from about 0.05% to about 0.5%, by weight of the suspension; (b) buffers such as citric acid-sodium citrate, phosphoric acid-sodium phosphate, and acetic acid-sodium acetate in amounts up to about 1%, and preferably from about 0.05% to about 0.5%, by weight of the suspension;
(c) suspending agents or thickeners such as cellulosics like methylcellulose, carrageenans like alginic acid and its derivatives, xanthan gums, gelatin, acacias, and microciystalline cellulose in amounts up to about 20%, and preferably from about 1% to about 15%, by weight of the suspension;
(d) antifoaming agents such as dimethyl polysiloxane in amounts up to about 0.2%, and preferably from about 0.01% to about 0.1%, by weight of the suspension; (e) sweetening agents such as those sweeteners well known in the art, including both natural and artificial sweeteners. Sweetening agents such as monosaccharides, disaccharides and polysaccharides such as xylose, ribose, glucose (dextrose), mannose, galactose, fructose (levulose), sucrose (sugar), maltose, invert sugar (a mixture of fructose and glucose derived from sucrose), partially hydrolyzed starch, com syrup solids, dihydrochalcones, monellin, steviosides, glycyrrhizin, and sugar alcohols such as sorbitol, mannitol, maltitol, hydrogenated starch hydrolysates and mixtures thereof may be utilized in amounts up to about 60%, and preferably from about 20% to about 50%, by weight of the suspension. Water-soluble artificial sweeteners such as soluble saccharin salts, i.e., sodium or calcium saccharin salts, cyclamate salts, the sodium, ammonium or calcium salt of 3,4-dihydro-6-methyl-l,2,3- oxathiazine-4-one-2,2-dioxide, the potassium salt of 3,4-dihydro-6-methyl- 1,2,3 - oxathiazine-4-one-2,2-dioxide (Acesulfame-K), the free acid form of saccharin, and the like may be utilized in amounts from about 0.001% to about 5%, by weight of the suspension;
(f flavoring agents such as those flavors well known to the skilled artisan, such as natural and artificial flavors and mints, such as peppermint, menthol, citrus flavors such as orange and lemon, artificial vanilla, cinnamon, various fruit flavors, both individual and mixed and die like may be utilized in amounts from about 0.5% to about 5%, by weight of the suspension;
(g) coloring agents such as pigments which may be incorporated in amounts up to about 6%, by weight of the suspension. A prefeπed pigment, titanium dioxide, may be incorporated in amounts up to about 2%, and preferably less than about 1%, by weight of the suspension. The coloring agents may also include natural food colors and dyes suitable for food, drug and cosmetic applications. These colorants are known as F.D.& C. dyes and lakes. The materials acceptable for the foregoing uses are preferably water-soluble. Such dyes are generally present in amounts up to about 0.25%, and preferably from about 0.05% to about 0.2%, by weight of the suspension;
(h) decolorizing agents such as sodium metabisulfite, ascorbic acid and the like may be incorporated into the suspension to prevent color changes due to aging. In general, decolorizing agents may be used in amounts up to about 0.25%, and preferably from about 0.05% to about 0.2%, by weight of the suspension; and
(i) solubilizers such as alcohol, propylene glycol, polyethylene glycol, and die like may be used to solubilize the flavoring agents. In general, solubilizing agents may be used in amounts up to about 10%, and preferably from about 2% to about 5%, by weight of the suspension.
The pharmaceutical suspensions of the present invention may be prepared as follows:
(A) admix the thickener with water heated from about 40°C. to about 95°C, preferably from about 40°C. to about 70°C, to form a dispersion if the thickener is not water soluble or a solution if the thickener is water soluble;
(B) admix die sweetening agent witii water to form a solution; (C) admix the therapeutic wound healing composition with the diickener-water admixture to form a uniform thickener-therapeutic wound healing composition;
(D) combine the sweetener solution with the thickener-therapeutic wound healing composition and mix until uniform; and (E) admix the optional adjunct materials such as coloring agents, flavoring agents, decolorants, solubilizers, antifoaming agents, buffers and additional water with the mixture of step (D) to form the suspension.
The ingestible therapeutic wound healing compositions of this invention may also be in chewable form. To achieve acceptable stability and quality as well as good taste and mouth feel in a chewable formulation several considerations are important. These considerations include die amount of active substance per tablet, the flavoring agent employed, the degree of compressibility of the tablet and die organoleptic properties of the composition.
Chewable therapeutic candy is prepared by procedures similar to those used to make soft confectionery. In a typical procedure, a boiled sugar-corn syrup blend is formed to which is added a frappe mixture. The boiled sugar-corn syrup blend may be prepared from sugar and com syrup blended in parts by weight ratio of about 90:10 to about 10:90. The sugar-corn syrup blend is heated to temperatures above about 120°C. to remove water and to form a molten mass. The frappe is generally prepared from gelatin, egg albumin, milk proteins such as casein, and vegetable proteins such as soy protein, and the like, which is added to a gelatin solution and rapidly mixed at ambient temperature to form an aerated sponge like mass. The frappe is then added to the molten candy mass and mixed until homogeneous at temperatures between about 65°C. and about 120°C.
The ingestible therapeutic wound healing composition of the instant invention can then be added to the homogeneous mixture as the temperature is lowered to about 65°C.-95°C. whereupon additional ingredients can then be added such as flavoring agents and coloring agents. The formulation is further cooled and formed into pieces of desired dimensions.
A general discussion of the lozenge and chewable tablet forms of confectionery may be found in HA. Lieberman and L. Lachman, Pharmaceutical Dosage Forms: Tablets Volume 1. Marcel Dekker, Inc., New York, N.Y. at pages 289 to 466, which disclosure is incorporated herein by reference.
In accordance witii tiiis invention, therapeutically effective amounts of the therapeutic wound healing compositions of the present invention may be admixed into the hard and soft confectionery products. These amounts are readily determined by those skilled in the art without the need for undue experimentation. In a prefeπed embodiment, die ingestible therapeutic wound healing composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 10% and an ingestible vehicle, that is a pharmaceutically acceptable carrier, in a quantity sufficient to bring the total amount of composition to 100%, by weight the ingestible therapeutic wound healing composition. In a more prefeπed embodiment, the ingestible composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most prefeπed embodiment, the ingestible composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and an ingestible vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight the ingestible therapeutic wound healing composition. The present invention extends to metiiods of making the ingestible therapeutic wound healing compositions. In such methods, an ingestible therapeutic wound healing composition is prepared by admixing a therapeutically effective amount of the therapeutic wound healing composition with a pharmaceutically-acceptable carrier. The apparatus useful in accordance with the present invention comprises mixing and heating apparatus well known in the confectionery arts, and therefore the selection of the specific apparatus will be apparent to the artisan. The final ingestible therapeutic wound healing compositions are readily prepared using methods generally known in the confectionery arts.
The therapeutic wound healing compositions may also be incorporated into chewing gums. In this form of the invention, the chewing gum composition contains a gum base, a bulking agent, the inventive therapeutic wound healing composition, and various additives.
The gum base employed will vary greatly depending upon various factors such as the type of base desired, die consistency of gum desired and die odier components used in die composition to make the final chewing gum product. The gum base may be any water-insoluble gum base known in the art, and includes tiiose gum bases utilized for chewing gums and bubble gums. Illustrative examples of suitable polymers in gum bases include botii natural and synthetic elastomers and rubbers. For example, those polymers which are suitable as gum bases include, without limitation, substances of vegetable origin such as chicle, crown gum, nispero, rosadinha, jelutong, perillo, niger gutta, tunu, balata, gutta-percha, lechi-capsi, sorva, gutta kay, mixtures thereof and die like. Synthetic elastomers such as butadiene-styrene copolymers, polyisobutylene, isobutylene-isoprene copolymers, polyethylene, mixtures thereof and the like are particularly useful.
The gum base may include a non-toxic vinyl polymer, such as polyvinyl acetate and its partial hydrolysate, polyvinyl alcohol, and mixtures thereof. When utilized, die molecular weight of the vinyl polymer may range from about 2,000 up to and including about 94,000. The amount of gum base employed will vary greatly depending upon various factors such as the type of base used, die consistency of the gum desired and the other components used in the composition to make the final chewing gum product. In general, the gum base will be present in amounts from about 5% to about 94%, by weight of the final chewing gum composition, and preferably in amounts from about
15% to about 45%, and more preferably in amounts from about 15% to about 35%, and most preferably in amounts from about 20% to about 30%, by weight of the final chewing gum composition.
The gum base composition may contain conventional elastomer solvents to aid in softening the elastomer base component. Such elastomer solvents may comprise terpinene resins such as polymers of Alpha-pinene or β-pinene, methyl, glycerol or pentaerythritol esters of rosins or modified rosins and gums, such as hydrogenated, dimerized or polymerized rosins or mixtures thereof. Examples of elastomer solvents suitable for use herein include die pentaerythritol ester of partially hydrogenated wood or gum rosin, the pentaerythritol ester of wood or gum rosin, the glycerol ester of wood rosin, the glycerol ester of partially dimerized wood or gum rosin, the glycerol ester of polymerized wood or gum rosin, the glycerol ester of tall oil rosin, the glycerol ester of wood or gum rosin and the partially hydrogenated wood or gum rosin and die partially hydrogenated methyl ester of wood or rosin, mixtures thereof, and the like. The elastomer solvent may be employed in amounts from about 5% to about 75%, by weight of the gum base, and preferably from about 45% to about 70%, by weight of the gum base.
A variety of traditional ingredients may be included in the gum base in effective amounts such as plasticizers or softeners such as lanolin, palmitic acid, oleic acid, stearic acid, sodium stearate, potassium stearate, glyceryl triacetate, glyceryl lecithin, glyceryl monostearate, propylene glycol monostearate, acetylated monoglyceride, glycerine, mixtures thereof, and the like may also be incorporated into die gum base to obtain a variety of desirable textures and consistency properties.
Waxes, for example, natural and synthetic waxes, hydrogenated vegetable oils, petroleum waxes such as polyurethane waxes, polyethylene waxes, paraffin waxes, microcrystalline waxes, fatty waxes, sorbitan monostearate, tallow, propylene glycol, mixtures thereof, and the like may also be incorporated into the gum base to obtain a variety of desirable textures and consistency properties. These traditional additional materials are generally employed in amounts up to about 30%, by weight of the gum base, and preferably in amounts from about 3% to about 20%, by weight of die gum base.
The gum base may include effective amounts of mineral adjuvants such as calcium carbonate, magnesium carbonate, alumina, aluminum hydroxide, aluminum silicate, talc, tricalcium phosphate, dicalcium phosphate and die like as well as mixtures thereof. These mineral adjuvants may serve as fillers and textural agents. These fillers or adjuvants may be used in the gum base in various amounts. Preferably the amount of filler when used will be present in an amount up to about 60%, by weight of the chewing gum base.
The chewing gum base may additionally include the conventional additives of coloring agents, antioxidants, preservatives and die like. For example, titanium dioxide and other dyes suitable for food, drug and cosmetic applications, known as F.D. & C. dyes, may be utilized. An antioxidant such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, and mixtures thereof, may also be included. Other conventional chewing gum additives known to one having ordinary skill in the chewing gum art may also be used in die chewing gum base.
The gum composition may include effective amounts of conventional additives selected from the group consisting of sweetening agents (sweeteners), plasticizers, softeners, emulsifiers, waxes, fillers, bulking agents, mineral adjuvants, flavoring agents (flavors, flavorings), coloring agents (colorants, colorings), antioxidants, acidulants, thickeners, mixtures thereof and the like. Some of these additives may serve more than one purpose. For example, in sugarless gum compositions, the sweetener, e.g., sorbitol or other sugar alcohol or mixtures thereof, may also function as a bulking agent. Similarly, in sugar containing gum compositions, the sugar sweetener can also function as a bulking agent.
The plasticizers, softeners, mineral adjuvants, colorants, waxes and antioxidants discussed above as being suitable for use in the gum base may also be used in the gum composition. Examples of other conventional additives which may be used include emulsifiers, such as lecithin and glyceryl monostearate, thickeners, used alone or in combination with other softeners, such as methyl cellulose, alginates, caπageenan, xanthan gum, gelatin, carob, tragacanth, locust bean, and carboxy methyl cellulose, acidulants such as malic acid, adipic acid, citric acid, tartaric acid, fumaric acid, and mixtures thereof, and fillers, such as those discussed above under die category of mineral adjuvants. The fillers when used may be utilized in an amount up to about 60%, by weight of the gum composition.
Bulking agents (carriers, extenders) suitable for use in chewing gums include sweetening agents selected from the group consisting of monosaccharides, disaccharides, poly-saccharides, sugar alcohols, and mixtures thereof; polydextrose; maltodextrins; minerals, such as calcium carbonate, talc, titanium dioxide, dicalcium phosphate, and die like. Bulking agents may be used in amounts up to about 90%, by weight of the final gum composition, with amounts from about 40% to about 70%, by weight of the gum composition being prefeπed , with from about 50% to about 65%, by weight, being more prefeπed and from about 55% to about 60%, by weight of the chewing gum composition, being most preferred.
The sweetening agent used may be selected from a wide range of materials including water-soluble sweeteners, water-soluble artificial sweeteners, water- soluble sweeteners derived from naturally occurring water-soluble sweeteners, dipeptide based sweeteners, and protein based sweeteners, including mixtures thereof. Without being limited to particular sweeteners, representative categories and examples include: (a) water-soluble sweetening agents such as monosaccharides, disaccharides and polysaccharides such as xylose, ribulose, glucose (dextrose), mannose, galactose, fructose (levulose), sucrose (sugar), maltose, invert sugar (a mixture of fructose and glucose derived from sucrose), partially hydrolyzed starch, co syrup solids, dihydrochalcones, monellin, steviosides, glycyrrhizin, and sugar alcohols such as sorbitol, mannitol, maltitol, hydrogenated starch hydrolysates and mixtures thereof; (b) water-soluble artificial sweeteners such as soluble saccharin salts, i.e., sodium or calcium saccharin salts, cyclamate salts, the sodium, ammonium or calcium saltof3,4-dihydro-6-methyl-l ,2,3-oxathiazine-4-one-2,2-dioxide, the potassium salt of 3,4-dihydro-6-methyl-l,2,3-oxathiazine-4-one-2,2-dioxide (Acesulfame-K), die free acid form of saccharin, and die like; (c) dipeptide based sweeteners, such as L-aspartic acid derived sweeteners, such as L-aspartyl-L-phenylalanine methyl ester (Aspartame) and materials described in United States Patent No. 3,492,131, L-Alpha-aspartyl-N-(2,2,4,4- tetramethyl-3-tiιietanyl)-D-alanin-amide hydrate (Alitame), methyl esters of L-aspartyl- L-phenylglycerineandL-aspartyl-L-2,5-dihydrophenyl-glycine, L-aspartyl-2,5-dihydro- L-phenylalanine; L-aspartyl-L-(l-cyclohexen)-alanine, and the like;
(d) water-soluble sweeteners derived from naturally occurring water- soluble sweeteners, such as chlorinated derivatives of ordinary sugar (sucrose), known, for example, under die product designation of Sucralose; and
(e) protein based sweeteners such as tiiaumaoccous danielli (Thaumatin I and II).
In general, an effective amount of sweetener is utilized to provide die level of bulk and or sweetness desired, and this amount will vary with the sweetener selected. This amount of sweetener will normally be present in amounts from about 0.0025% to about 90%, by weight of the gum composition, depending upon die sweetener used. The exact range of amounts for each type of sweetener is well known in the art and is not the subject of the present invention. The amount of sweetener ordinarily necessary to achieve the desired level of sweetness is independent from die flavor level achieved from flavor oils.
Prefeπed sugar based-sweeteners are sugar (sucrose), com syrup and mixtures thereof. Prefeπed sugarless sweeteners are the sugar alcohols, artificial sweeteners, dipeptide based sweeteners and mixtures thereof. Preferably, sugar alcohols are used in die sugarless compositions because these sweeteners can be used in amounts which are sufficient to provide bulk as well as the desired level of sweetness. Prefeπed sugar alcohols are selected from the group consisting of sorbitol, xylitol, maltitol, mannitol, and mixtures thereof. More preferably, sorbitol or a mixture of sorbitol and mannitol is utilized. The gamma form of sorbitol is preferred. An artificial sweetener or dipeptide based sweetener is preferably added to the gum compositions which contain sugar alcohols.
The coloring agents useful in the gum compositions are used in amounts effective to produce the desired color. These coloring agents include pigments which may be incorporated in amounts up to about 6% by weight of the gum composition. A prefeπed pigment, titanium dioxide, may be incorporated in amounts up to about 2%, and preferably less than about 1% by weight of the composition. The colorants may also include natural food colors and dyes suitable for food, drug and cosmetic applications. These colorants are known as F.D.& C. dyes and lakes. The materials acceptable for the foregoing uses are preferably water-soluble. Illustrative nonlimiting examples include die indigoid dye known as F.D.& C. Blue No.2, which is the disodium salt of 5,5-indigotindisulfonic acid. Similarly, the dye known as F.D.& C. Green No.l comprises a triphenylmethane dye and is the monosodium salt of 4-[4-(N- ethyl-p-sulfoniumbe-izyl--mino)liphenylmethylene]-[l-(N-ethyl-N-p-sulfoniumbenzyl)- delta-2,5-cyclohexadieneimine]. A full recitation of all F.D.& C. colorants and their coπesponding chemical structures may be found in the Kirk-Othmer Encyclopedia of Chemical Technology. 3rd Edition, in volume 5 at pages 857-884, which text is incorporated herein by reference.
Suitable oils and fats usable in gum compositions include partially hydrogenated vegetable or animal fats, such as coconut oil, palm kernel oil, beef tallow, lard, and the like. These ingredients when used are generally present in amounts up to about 7%, by weight, and preferably up to about 3.5%, by weight of the gum composition. In accordance witii this invention, therapeutically effective amounts of the therapeutic wound healing compositions of the present invention may be admixed into a chewing gum. These amounts are readily determined by tiiose skilled in the art without the need for undue experimentation. In a prefeπed embodiment, die final chewing gum composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 10% and a chewing gum composition in a quantity sufficient to bring die total amount of composition to 100%, by weight of the chewing gum composition. In a more prefeπed embodiment, die final chewing gum composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most preferred embodiment, the final chewing gum composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and a chewing gum composition in a quantity sufficient to bring the total amount of composition to 100%, by weight of the chewing gum composition.
The present invention extends to metiiods of making the therapeutic chewing gum compositions. The therapeutic wound healing compositions may be incorporated into an otiierwise conventional chewing gum composition using standard techniques and equipment known to those skilled in the art. The apparatus useful in accordance with the present invention comprises mixing and heating apparatus well known in the chewing gum manufacturing arts, and therefore the selection of the specific apparatus will be apparent to the artisan.
For example, a gum base is heated to a temperature sufficiently high enough to soften the base without adversely effecting the physical and chemical make up of the base. The optimum temperatures utilized may vary depending upon die composition of the gum base used, but such temperatures are readily determined by tiiose skilled in the art without undue experimentation.
The gum base is conventionally melted at temperatures that range from about 60°C. to about 120°C. for a period of time sufficient to render the base molten. For example, the gum base may be heated under these conditions for a period of about thirty minutes just prior to being admixed incrementally with the remaining ingredients of the base such as the plasticizer, fillers, the bulking agent and/or sweeteners, die softener and coloring agents to plasticize the blend as well as to modulate die hardness, viscoelasticity and formability of the base. The chewing gum base is then blended witii the therapeutic wound healing composition of the present invention which may have been previously blended witii other traditional ingredients. Mixing is continued until a uniform mixture of gum composition is obtained. Thereafter the gum composition mixture may be formed into desirable chewing gum shapes.
In a specific embodiment, die invention is directed to a therapeutic pharmaceutical composition for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises:
(A) a therapeutically effective amount of a therapeutic wound healing composition of Embodiment One (I) selected from the group consisting of:
(IA) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein die fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
(LB) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; (LC) (a) an antioxidant; and
(b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are tiiose fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
(LD) (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(B) a pharmaceutically acceptable carrier.
The pharmaceutically acceptable carrier may be selected from die group consisting of pharmaceutical appliances, topical vehicles, and ingestible vehicle.
In another specific embodiment, the invention is directed to a method for preparing a therapeutic pharmaceutical composition for preventing and reducing injury to mammalian cells, and increasing the resuscitation rate of injured mammalian cells, which comprises the steps of :
(A) providing a therapeutically effective amount of a therapeutic wound healing composition of Embodiment One (LA-D) selected from the group consisting of:
(I A) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
(LB) (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
( C) (a) an antioxidant; and
(b) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are tiiose fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
(I.D) (a) lactate selected from the group consisting of lactic acid, pharmaceutically acceptable salts of lactic acid, and mixtures thereof;
(b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are tiiose fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(B) providing a pharmaceutically acceptable carrier; and
(C) admixing the therapeutic wound healing composition from step (A) and die pharmaceutically acceptable carrier from step (B) to form a therapeutic pharmaceutical composition.
Throughout this application, various publications have been referenced. The disclosures in these publications are incorporated herein by reference in order to more fully describe the state of the art.
The present invention is further illustrated by d e following examples which are not intended to limit the effective scope of the claims. All parts and percentages in the examples and throughout the specification and claims are by weight of die final composition unless otherwise specified. E. Examples Of
The Therapeutic Wound Healing Compositions
Of Embodiment One (LA-D)
Study 1
This study demonstrates a comparison of the viability of U937 monocytic cells after exposure of the cells to various antioxidants and combinations of antioxidants. This study also demonstrate a comparison of the levels of hydrogen peroxide produced by U937 monocytic cells and mammalian epidermal keratinocytes after exposure of the cells to various antioxidants and combinations of antioxidants.
The results of this study are illustrated in Figures 1-4 and examples 1-26 below.
Mammalian epidermal keratinocytes and monocytes were employed to examine the ability of various antioxidants to reduce levels of hydrogen peroxide in tiiese cells. Hydrogen peroxide was measured after die cells were exposed to ultraviolet light in the wavelength range from 290 to 320 nm (UV-B) or to the inflammatory compound 12-0-tetradecanoyl-phorbol-13-acetate (TPA). Sodium pyruvate was tested at various concentrations to determine the effect of concentrations of this antioxidant on the hydrogen peroxide production by epidermal cells and monocytes. Magnesium pyruvate, calcium pyruvate, zinc pyruvate, and combinations of sodium pyruvate with ascorbic acid, lactic acid, and Vitamin E were then tested to determine the effect of these salts and combinations of antioxidants on the hydrogen peroxide production by epidermal cells and monocytes.
Mammalian epidermal keratinocytes were isolated by trypsin-zation of epithelial sheets and grown in modified basal MCDB 153 medium supplemented witii epidermal growth factor, bovine pituitary extract, and hydrocortisone. Cells were maintained in a humidified incubator witii 5% carbon dioxide at 37°C. Keratinocytes were seeded in 60 mm culture dishes at a cell density of 3 x 10 cells per dish and die cultures were exposed to 1 M.E.D. dose of ultraviolet-B light (100 mJ/cm ) or treated with lOO ng/ml of TPA. U937 monocytic cells are a cultured cell line grown in RPMI media with 10% fetal calf serum. Cells were maintained in a 60 mm culture dish at 5% carbon dioxide at 37°C. at a seeding density not exceeding 1 x 106 cells per dish.
Sodium pyruvate, lactic acid, ascorbic acid, and Vitamin E were dissolved in distilled water, with sufficient surfactant. The concentrations of the sodium pyruvate solutions prepared were 1 mM, 10 mM, 50 mM, 100 mM, and 200 mM. The concentrations of the lactic acid solutions prepared were 1.0%, 0.1%, and 0.05%. The concentrations of the ascorbic acid solutions prepared were 1.0%, 0.1%, 0.05%, and 0.025%. The concentrations of the Vitamin E solutions prepared were 1 U, 10 U, 50 U, and 100 U. The test solutions were adjusted to a pH value of 7.4 with 1.0N sodium hydroxide solution and then sterile filtered. The appropriate concentration of test solution or combination of test solutions was added to die cells immediately prior to exposure of the cells to ultraviolet light-B or TPA [100ng/ml]. Stock solutions were prepared so tiiat the vehicle did not constitute more than 1% of the total volume of the culture media.
Intracellular hydrogen peroxide production by mammalian epidermal keratinocytes and U937 monocytes was measured using dichlorofluorescein diacetate (DCFH-DA, Molecular Probes, Eugene, Ore.). DCFH-DA is a non-polar non- fluorescent compound that readily diffuses into cells where it is hydrolyzed to the polar non-fluorescent derivative DCFH which then becomes trapped within the cells. In the presence of intracellular hydrogen peroxide, DCFH is oxidized to die highly fluorescent compound DCF. Hence, cellular fluorescence intensity is directly proportional to the level of intracellular hydrogen peroxide produced. Cellular fluorescence intensity can be monitored by fluorimetry and by flow cytometry.
Mammalian epidermal keratinocytes and U937 cultured monocytes (1 x 106 per dish) were incubated at 37°C. with 5 uM of DCFH-DA. Production of hydrogen peroxide was measured using a Coulter Profile analytical flow cytometer.
Linear and log intensity of green fluorescence data was collected. For each analysis, a quantity of 10,000 to 20,000 events was accumulated. Optical alignment for the instrument was performed daily. Coefficients of variation for forward angle light scatter and integrated green fluorescence were generally less than two. Each analysis was repeated three times and the quantitation of fluorescence was expressed in terms of femtomoles (fmol, 10"1-5 moles) of DCF oxidized per cell, which is a direct measure of the intracellular hydrogen peroxide produced. Alternatively, in the saturated and unsaturated fatty acid examples in examples 27-52, fluorimetry was used to assess die DCF oxidation per cell.
The viability of the U937 monocytic cells after exposure of the cells to various antioxidants for 24 hours was measured. The viability of the cells was determined by exposing the cells to the dye propidium iodide. Permeable cell membranes which absorbed die dye were not considered viable. The viability of the cells was represented as die percentage of cells that excluded propidium iodide. Figure 1 depicts in bar graph format the viability of U937 monocytic cells after exposure of the cells to no antioxidant (Example 1, control), to sodium pyruvate
(Example 2), to ascorbic acid (Example 3), to lactic acid (Example 4), and to Vitamin E (Example 5). Figure 2 depicts in bar graph format the viability of U937 monocytic cells after exposure of the cells to various combinations of antioxidants. Specifically, the viability of U937 monocytic cells was measured after exposure to no antioxidant (Example 6, control), to ascorbic acid and lactic acid (Example 7), to ascorbic acid and Vitamin E (Example 8), to sodium pyruvate and ascorbic acid (Example 9), to sodium pyruvate and lactic acid (Example 10), to sodium pyruvate and Vitamin E (Example 11), to lactic acid and Vitamin E (Example 12), and to sodium pyruvate, ascorbic acid, and lactic acid (Example 13).
Figure 1 shows that ascorbic acid is cytotoxic to monocytes at concentrations as low as 0.25%. Figure 2 shows that the cytotoxicity of ascorbic acid was reversed by the addition of 10 mM of sodium pyruvate. Figures 1 and 2 show that the viability rate of 15% to 20% of the cells when treated with ascorbic acid was increased to 95% to 98% upon addition of sodium pyruvate. Lactic acid and
Vitamin E did not reverse the cytotoxicity of ascorbic acid. Sodium pyruvate was then tested at various concentrations to determine die effect of concentrations of this antioxidant on the hydrogen peroxide production by epidermal cells and monocytes. Mammalian epidermal keratinocytes and monocytes were exposed to (a) 1 M.E.D. dose of ultraviolet light-B and (b) 100 ng/ml of 12-O- tetradecanoylphorbol-13-acetate (TPA) in the presence of sodium pyruvate at the following concentrations: 200 mM, 100 mM, 50 mM, 10 mM, 1 mM.
The optimum concentration of sodium pyruvate to reduce the hydrogen peroxide production by epidermal cells and monocytes was found to be 10 mM. Concentrations of sodium pyruvate of 50 mM and above were cytotoxic to both epidermal keratinocytes and monocytes.
Magnesium pyruvate, calcium pyruvate, zinc pyruvate, ascorbic acid, lactic acid, and Vitamin E, and combinations of sodium pyruvate with ascorbic acid, lactic acid, and Vitamin E were then tested to determine the effect of these salts and combinations of antioxidants on die hydrogen peroxide production by epidermal cells and monocytes. The following test solutions were prepared.
(a) sodium pyruvate [10 mM]; (b) zinc salt [10 mM];
(c) magnesium salt [10 mM];
(d) calcium salt [10 mM];
(e) sodium pyruvate [10 mM] and ascorbic acid [0.025%]; (f) sodium pyruvate [10 mM] and lactic acid
[0.05%];
(g) sodium pyruvate [10 mM], lactic acid, [0.05%], and ascorbic acid [0.025%];
(h) lactic acid [1.0%, 0.1%, and 0.05%]; (i) ascorbic acid [1.0%, 0.1%, 0.05%, and
0.025%];
(j) Vitamin E [1 U, 10 U, 50 U, and 100 U]; and (k) vehicle solvent controls.
There was no significant difference among the zinc, magnesium, and calcium salts of pyruvic acid on die hydrogen peroxide production by epidermal cells and monocytes. The zinc and calcium salts of pyruvic acid induced differentiation of keratinocytes. For convenience, die sodium salt was used in subsequent tests.
The optimum concentration of lactic acid to reduce the hydrogen peroxide production by epidermal cells and monocytes was found to be 0.05%. The optimum concentration of ascorbic acid was found to be 0.025%. The higher concentrations of both of these compounds were found to be cytotoxic to both types of cells. The optimum concentration of Vitamin E was found to be 50 U.
Figure 3 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells after exposure of the cells to no antioxidant
(Example 14, control), to sodium pyruvate (Example 15), to ascorbic acid (Example 16), to lactic acid (Example 17), and to Vitamin E (Example 18). Sodium pyruvate and Vitamin E significantly reduced die hydrogen peroxide production by monocytes.
Figure 4 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells after exposure of the cells to various combinations of antioxidants. Specifically, the levels of hydrogen peroxide produced by U937 monocytic cells were measured after exposure to no antioxidant (Example 19, control), to ascorbic acid and lactic acid (Example 20), to ascorbic acid and Vitamin E
(Example 21), to sodium pyruvate and ascorbic acid (Example 22), to sodium pyruvate and lactic acid (Example 23), to sodium pyruvate and Vitamin E (Example 24), to lactic acid and Vitamin E (Example 25), and to sodium pyruvate, ascorbic acid, and lactic acid (Example 26). The combination of lactic acid (0.05%) and Vitamin E (50 U) significantly reduced the hydrogen peroxide production by monocytes. The morphological alterations in epidermal keratinocytes were observed in control cultures and in cultures exposed to ultraviolet-B. Cells in the layer closest to the dermis are basal keratinocytes. These cells proliferate and migrate into the spinous and granular layers of the epidermis where die cells begin to differentiate. The differentiation pattern results in cells enucleating and forming comified envelopes at the uppermost portion of the epidermis, the statum corneum. The differentiation of keratinocytes is controlled by the levels of calcium, magnesium, and otiier elements in the medium. Cells in culture systems promoting differentiation appear as an epidermal sheet forming attachments or tight junctions with each other. Keratinocytes that become nonadherent or float in the media were considered responding to a cytotoxic event.
The following morphological alterations in the mammalian epidermal keratinocytes were observed for the following control cultures:
10 mM Sodium Pyruvate: Tight junctions of cells were formed and die proliferation rate of the cells was higher than the rate of the control cells.
0.025% Ascorbic Acid: Cells were floating in a cytotoxic response to ascorbic acid.
0.025% Ascorbic acid and 10 mM Sodium Pyruvate: Few tight junctions of cells were observed and cells appeared similar to the cells in the sodium pyruvate culture.
0.05% Lactic Acid: Cells appeared dramatically altered as an epidermal sheet and as flat granular cells.
0.05% Lactic Acid and 10 mM Sodium Pyruvate: Cells formed an epidermal sheet but appeared smaller than the cell in the lactic acid culture.
50 U Vitamin E: Cells appeared the same as the cells in the control culture. 50 U Vitamin E and 10 mM Sodium Pyruvate: Cells increased in number and changed in appearance resembling the cells in the sodium pyruvate culture.
The following morphological alterations in the mammalian epidermal keratinocytes were observed for the coπesponding cultures exposed to ultraviolet light- B, 100 mJoules, for 24 hours:
10 mM Sodium Pyruvate: Cells proliferated more rapidly than the cells in the control culture.
0.025% Ascorbic Acid: Cells were nonadherent and floating in a cytotoxic response to ascorbic acid greater than the cytotoxic response of the coπesponding cells without ultraviolet-B light exposure.
0.05% Lactic Acid: Cells formed an epidermal sheet and were more granular than cells in the control culture without ultraviolet-B light exposure.
50 U Vitamin E: Cell growth was inhibited but cells appeared similar to cells in the control culture without ultraviolet-B light exposure.
50 U Vitamin E and 10 mM Sodium Pvmvate: Cells appeared similar to cells in the control culture and proliferated to a greater extent than cells in the control cultures without ultraviolet-B light exposure.
Morphological alterations in the U937 monocytic cell line were also observed for control cultures and cultures exposed to ultraviolet light-B, 100 mJoules, for 24 hours. The following compounds and combination of compounds, at die concentrations set out below, significantly inhibited the levels of hydrogen peroxide produced by U937 monocytic cells Sodium pyruvate at 10 mM and 50 mM;
Vitamin E at 50 U and 100 U; and
Lactic acid at 0.05% and Vitamin E at 50 U.
Examples Of The Therapeutic Wound Healing Compositions Of Embodiment One (I.A-D) Study 2
This study demonstrates a comparison of the levels of hydrogen peroxide produced by U937 monocytic cells and epidermal keratinocytes after exposure of the cells to various combinations of antioxidants witii and without a mixture of saturated and unsaturated fatty acids. The results of this study are illustrated in Figures 5-7 and examples 27-52 below.
Mammalian epidermal keratinocytes and U937 monocytic cells and the test solutions of sodium pyruvate, lactic acid, ascorbic acid, and Vitamin E were prepared as describe above for Examples 1-26. Intracellular hydrogen peroxide production by the mammalian epidermal keratinocytes and U937 monocytes was also measured as described above.
A mixture of fatty acids derived from chicken fat was prepared for addition to the cultured cells by mixing 0.1% of the chicken fat with the culture media. At the temperature of the culture media, 37°C, the chicken fat was miscible. This chicken fat mixture was added to cultures of cells prior to exposure of the cells to ultraviolet-B light or TPA treatment.
As set out in examples 1-26, mammalian epidermal keratinocytes and monocytes were exposed to (a) 1 M.E.D. dose of ultraviolet light-B and (b) 100 ng/ml of 12-O-tetradecanoylphorbol-13-acetate in the presence of various antioxidants and combinations of antioxidants with and witiiout a mixture of saturated and unsaturated fatty acids [0.1%, 0.5%, and 1.0% chicken fat].
Figure 5 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells after exposure of the cells to various combinations of antioxidants with and without a mixture of saturated and unsaturated fatty acids. Specifically, die levels of hydrogen peroxide produced by U937 monocytic cells were measured after exposure to lactic acid and Vitamin E without fatty acids (Example 27) and with fatty acids (Example 28), to ascorbic acid and lactic acid without fatty acids (Example 29) and witii fatty acids (Example 30), and to ascorbic acid and Vitamin E witiiout fatty acids (Example 31) and witii fatty acids (Example 32). The ability of the combinations of lactic acid and Vitamin E, ascorbic acid and lactic acid, and ascorbic acid and Vitamin E to reduce the hydrogen peroxide production by monocytes was increased in the presence of fatty acids. The most effective combination to reduce the hydrogen peroxide production of monocytes was lactic acid (0.05%) and Vitamin E
(50 E) in the presence of a mixture of saturated and unsaturated fatty acids (0.5%).
Figure 6 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal keratinocytes after exposure of the cells to various antioxidants with and witiiout a mixture of saturated and unsaturated fatty acids. Specifically, the levels of hydrogen peroxide produced by epidermal keratinocytes were measured after exposure to no antioxidant witiiout fatty acids (Example 33, control) and witii fatty acids (Example 34), to sodium pyruvate without fatty acids (Example 35) and witii fatty acids (Example 36), to ascorbic acid witiiout fatty acids (Example 37) and witii fatty acids (Example 38), to lactic acid without fatty acids (Example 39) and witii fatty acids (Example 40), and to Vitamin E without fatty acids (Example 41) and witii fatty acids (Example 42). The ability of sodium pyruvate and Vitamin E to reduce the hydrogen peroxide production by epidermal keratinocytes was increased in the presence of fatty acids. The most effective combinations to reduce die hydrogen peroxide production of epidermal keratinocytes were sodium pyruvate in combination with a mixture saturated and unsaturated fatty acids and Vitamin E in combination with a mixture of saturated and unsaturated fatty acids. Figure 7 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal keratinocytes after exposure of the cells to various combinations of antioxidants with and witiiout a mixture of saturated and unsaturated fatty acids. Specifically, die levels of hydrogen peroxide produced by epidermal keratinocytes were measured after exposure to no antioxidant witiiout fatty acids (Example 43, control) and witii fatty acids (Example 44), to sodium pyruvate and ascorbic acid without fatty acids (Example 45) and witii fatty acids (Example 46), to sodium pyruvate and lactic acid witiiout fatty acids (Example 47) and witii fatty acids (Example 48), to sodium pyruvate and Vitamin E witiiout fatty acids (Example 49) and with fatty acids (Example 50), and to ascorbic acid and Vitamin E without fatty acids (Example 51) and witii fatty acids (Example 52). The ability of all combinations of antioxidants to reduce the hydrogen peroxide production by epidermal keratinocytes was increased in the presence of fatty acids. In order of potency, the most effective combinations to reduce die hydrogen peroxide production of epidermal keratinocytes were sodium pyruvate and Vitamin E, sodium pyruvate and lactic acid, and Vitamin E, each in combination with a mixture of saturated and unsaturated fatty acids (0.5%).
Because of die cytotoxicity of cells towards ascorbic acid described above, die ascorbic acid combinations without sodium pyruvate were not considered significantly different from the control test solution.
Summary Analysis Of The Data From Studies 1 and 2
Human epidermal keratinocytes were isolated by trypsinization of epithelial sheets and grown in modified base MCDB 153 medium supplemented witii epidermal growth factor and bovine pituitary extract. Cells were seeded in culture dishes at a density of 3 x 105/dish. Prior to exposure to UV B light (100mJ/cm2) or treatment with lOOng/ml TPA, die cultures were treated with the appropriate concentration of wound healing components. Intracellular production of hydrogen peroxide was measured using DCFH-DA, a nonpolar compound that readily diffuses into cells, hydrolyzed to a nonpolar derivative. In the presence of intracellular hydrogen peroxide, DCFH is oxidized to a highly fluorescent compound DCF. Thus, cellular fluorescence intensity is directly proportional to levels of hydrogen peroxide produced and can be monitored by flow cytometry. Hydrogen peroxide is cytotoxic, therefore lower levels of hydrogen peroxide production is desirable for cellular viability.
In all cases, the three component wound healing composition surpassed die predicted outcomes, clearly demonstrating unpredicted synergy.
Results 1 2 3 4
1 - Control 250 250 0
2 - Fatty Acids 250 230 -20 (0.5%) 3 3 - - SSooddiiuumm PPyyrruuvvaattee 2 25500 490 +240 (lOmM)
4 - Vitamin E 250 400 +150 (50 units)
5 - Pyruvate & 250 430 +180 Fatty Acids
6 - Vitamin E & 250 200 -50 Fatty Acids
7 - Pyruvate & 250 290 +40 Vitamin E 8 8 - - P Pyyrruuvvaattee && 2 25500 120 -130
Vitamin E & Fatty Acids
Column 1 shows the different treatment groups. Column 2 shows the production of H202 in control cells (fmol/cell). Column 3 shows the production of H 0 after treatment with wound healing components. Column 4 shows the difference in production of H202 from control after the treatment.
All comparisons were assessed against the controls, which produced 250 H202 fmol/cell. The positive numbers represent H202 production in excess of the control and the negative numbers represent H202 production below the control. These results are set out in Figure 8.
Combination of Single Ingredient Effects
Fatty Acids (-20) & Vitamin E (+150) & Pyruvate (+240) +370 Is The Predicted Three Component Effect
-130 Is The Wound healing composition Actual Effect 500 Is The Difference Between Predicted Effect minus Actual effect
(Synergy)
Combination of Paired and Single Ingredients
Pyruvate & Fatty Acids (+ 180) & vitamin E (+ 150)
+330 Is The Predicted Three Component Effect
-130 Is The Wound healing composition Actual Effect
460 Is The Difference between Predicted Effect minus Actual Effect (Synergy)
Vitamin E & Fatty Acids (-50) & Pyruvate (+240)
+ 190 Is The Predicted Three Component Effect -130 Is The Wound healing composition Actual Effect 320 Is The Difference between Predicted Effect minus Actual Effect
(Synergy)
Pyruvate & Vitamin E (+40) & Fatty Acids (-20)
+20 Is The Predicted Three Component Effect -130 Is The Wound healing composition Actual Effect
150 Is The Difference between Predicted Effect minus Actual Effect
(Synergy) In all cases, the three component wound healing composition surpassed die predicted outcomes clearly demonstrating unpredicted synergy.
Examples Of
The Therapeutic Wound Healing Compositions
Of Embodiment One (I.A-D)
Study 3
This study demonstrates a comparison of the wound healing abilities of the therapeutic wound healing compositions of the present invention versus conventional wound healing compositions. The results of this study are illustrated in examples A-D.
The wound healing compositions of Examples A-D were prepared having the compositions set out in Table A.
Examples
Ingredient A B C D Prep-H™ sodium pyrubate — 2% — — vitamin E — 1% — — chicken fat — 2% — —
LYCD 2000 U* 2400 U 2400 U — shark liver oil 3%* 3% 3% — petrolatum in 64% 66.5% 68% mineral oil amounts 22.53% 25.03% 26.8% paraffin totaling 5% 5% 5% emulsifier 100% 0.2% 0.2% 0.2%
These components are present in Preparation H Wound healing composition A was commercially available Preparation H . Wound healing composition B was a petrolatum base formulation containing live yeast cell derivative, shark oil, and a mixture of sodium pyruvate, vitamin E, and chicken fat. Wound healing composition C was a petrolatum base formulation containing live yeast cell derivative and shark oil. Wound healing composition D was a petrolatum base formulation only.
Wound healing studies were carried out using hairless mice (SKR-1, Charles River) 6-8 weeks in age. One group of mice were untreated as a control group and were refeπed to as Example E. In each group there were 6 mice for evaluation at either day 3 or day 7 for a total number of 60 animals in the study. The mice were anesthetized with ether and a midline 3 cm full thickness longitudinal incision was made witii a number 10 scalpel blade. Incisions were closed using steel clips at 1 cm intervals. Formulations A-D set out above were applied in a randomized blinded study to die wounds on day 0 at 2 hours following wounding and reapplied at 24 hour intervals during the 7 days of die study. The wounds were examined daily and scored on a basis of 0-5 for closure on each day of die study, with a score of 5 representing the wound best healed.
The animals were sacrificed on day 3 and day 7 using cervical dislocation. The dorsal skin including the incision was dissected without the subcutaneous tissue. The skin was placed in neutral buffered formalin and subsequently sectioned and stained with hematoxylin and eosin. The wounds were examined microscopically and representative tissue sections were photographed.
On each day of die experiment, the score and rank order of die formulations for closure of wounds and speed of healing were as follows:
B (5) » D (4) » C (2) >/= E, Control (2) > A (1)
Photographs of the wounded mice on day 4 are set out in Figures 9 and 10. Figures 9 and 10 show tiiat Formulation B, which was a petrolatum base formulation containing live yeast cell derivative, shark oil, and a mixture of sodium pyruvate, vitamin E, and chicken fat, was a significantly better wound healing agent than the other formulations. These results are supported by die subjective grading of die wound closures and die speed of healing on each day (1-7) of the experiment as well as on the objective histological examination of tissue sections to measure the extent of inflammatory cell infiltrate within the wound and die extent of epithelialization at the wound edges. The final result was that less scar tissue was present at day 7 on the mice treated witii Formulation B.
Formulation D, which was a white petrolatum formulation only, was judged to be significantly more effective to promote healing than either Formulation C, which was a petrolatum base formulation containing shark liver oil and live yeast cell derivative, or Formulation A, which was Preparation H . The superior ability of Formulation D over Formulation C to improve healing may result from a delay in die healing process caused when the live yeast cell derivative is depleted and die cells shift to an alternative nutrient source. The presence of the mixture of sodium pyruvate, vitamin E, and chicken fat in Formulation B apparently offsets the depletion of the live yeast cell derivative.
Formulation C, which was a petrolatum base formulation containing live yeast cell derivative and shark oil, was judged comparable to the control (untreated wound) in speed of wound closure and extent of healing. Formulation A, which was Preparation H™, appeared to be die least effective healing formulation by both subjective grading of wound healing and by objective examination of tissue sections.
The superior ability of Formulation D and Formulation C over Formulation A to improve healing may be due to their ability to act as an occlusive wound dressing tiiat prevents transepidermal water loss and thus promotes healing and wound closure. The poor ability of Formulation A to improve healing may be due to the potential cytotoxicity of phenylmercuric nitrate present in Preparation H as a preservative. These results show that the wound healing compositions of the present invention which comprise a mixture of sodium pyruvate, vitamin E, and chicken fat increase the proliferation and resuscitation rate of mammalian cells. The wound healing compositions mediate low levels of oxygen in the initial stages of healing to suppress oxidative damage and higher levels of oxygen in die later stages of healing to promote collagen formation.
H. Sunscreen Wound Healing Compositions A. Embodiment Two (I.A-D + X)
Applicant has discovered tiierapeutic sunscreen-wound healing compositions (I.A-D + X) which comprise a therapeutically effective amount of a sunscreen agent and an anti-inflammatory agent (collectively refeπed to as X) and the wound healing compositions of Embodiment One (I A-D). Preferably, the wound healing composition (LA) comprises (a) pyruvate, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids. Sunscreen agents can help prevent sunburn by screening ultra violet light but do not heal injured mammalian cells. Anti- inflammatory agents can reduce inflammation (erythema) in a patient but do not promote the wound healing process. Wound healing compositions can increase the resuscitation rate of injured mammalian cells and die proliferation rate of new mammalian cells to replace dead cells. Wound healing compositions can also minimize oxygen radical damage from ultra violet light. Applicants have found tiiat the combination of a sunscreen agent, an anti-inflammatory agent, and a wound healing composition results in a therapeutic sunscreen-wound healing compositions useful for minimizing and treating sunburn damage. The sunscreen- wound healing compositions may optionally contain a therapeutically effective amount of a topical anesthetic to further reduce the duration and severity of sunburn.
The combination of the sunscreen agent, the anti-inflammatory agent, and die wound healing compositions of the present invention provides a pharmaceutical composition useful for minimizing and treating sunburn damage and having an enhanced ability to prevent and reduce injury to mammalian cells and further increase the resuscitation rate of injured mammalian cells. The tissue damage associated witii sunburn is believed to be caused by die production of cellular produced active oxygen species. Combination of the sunscreen agent, anti-inflammatory agent, and die wound healing compositions helps suppress such reactive oxygen-linked tissue injury.
Sunscreen agents are compounds which provide broad spectrum protection from ultra violet A and ultra violet B light from die sun. The sunscreen agents in the sunscreen-wound healing compositions of the present invention may be selected from a wide range of tiierapeutic agents and mixtures of therapeutic agents. Nonlimiting illustrative specific examples of sunscreen agents include etiiylhexyl p- methoxycinnamate, octyl methoxycinnamate, octyl dimethyl »-aminobenzoic acid, 2- ethylhexyl salicylate, octyl salicylate, menthyl anthranilate, octocrylene, padimate o, titanium dioxide, urea, and oxybenzone. Preferably, the sunscreen agent is oxybenzone.
The amount of sunscreen agent used in die present invention is a therapeutically effective amount and may vary depending upon the therapeutic dosage recommended or permitted for the particular sunscreen agent. In general, the amount of sunscreen agent present is the ordinary dosage required to obtain the desired result. Such dosages are known to the skilled practitioner in the medical arts and are not a part of the present invention. In a prefeπed embodiment, the sunscreen agent in the sunscreen-wound healing composition is present in an amount from about 1 % to about 30%, preferably from about 2% to about 25%, and more preferably from about 4% to about 20%, by weight.
Anti-inflammatory agents are compounds that counteract or suppress the inflammatory process. The anti-inflammatory agents in the sunscreen-wound healing compositions of the present invention may be selected from a wide variety of steroidal, non-steroidal, and salicylate water-soluble and water-insoluble drugs and their acid addition or metallic salts. Both organic and inorganic salts may be used provided the anti-inflammatory agent maintains its medicament value. The anti-inflammatory agents may be selected from a wide range of therapeutic agents and mixtures of therapeutic agents which may be administered in sustained release or prolonged action form. Nonlimiting illustrative specific examples of non-steroidal anti-inflammatory agents include the following medicaments: ibuprofen, naproxen, sulindac, diflunisal, piroxicam, indomethacin, etodolac, meclofenamate sodium, fenoproben calcium, ketoprofen, mefenamic acid, nabumetone, ketorolac tromethamine, diclofenac, and evening primrose oil (containing about 72% linoleic acid and about 9% gamma- linolenic acid). Nonlimiting illustrative specific examples of salicylate anti- inflammatory agents include die following medicaments: acetylsalicylic acid, mesalamine, salsalate, diflunisal, salicylsalicylic acid, and choline magnesium trisalicylate. Nonlimiting illustrative specific examples of steroidal anti-inflammatory agents include die following medicaments: flunisolide, triamcinoline, triamcinoline acetonide, beclometiiasone diproprionate, betamethasone diproprionate, hydrocortisone, cortisone, dexamethasone, predinisone, methyl prednisolone, and prednisolone.
Prefeπed anti-inflammatory agents to be employed may be selected from the group consisting of ibuprofen, naproxen, sulindac, diflunisal, piroxicam, indomethacin, etodolac, meclofenamate sodium, fenoproben calcium, ketoprofen, mefenamic acid, nabumetone, ketorolac tromethamine, diclofenac, evening primrose oil, acetylsalicylic acid, mesalamine, salsalate, diflunisal, salicylsalicylic acid, choline magnesium trisalicylate, flunisolide, triamcinoline, triamcinoline acetonide, beclometiiasone diproprionate, betamethasone diproprionate, hydrocortisone, cortisone, dexamethasone, predinisone, methyl prednisolone, and prednisolone. In a preferred embodiment, die anti-inflammatory agent is selected from the group consisting of ibuprofen, naproxen, sulindac, diflunisal, piroxicam, indomethacin, etodolac, meclofenamate sodium, fenoproben calcium, ketoprofen, mefenamic acid, nabumetone, ketorolac tromethamine, diclofenac, and evening primrose oil. In a more preferred embodiment, the - ti-infl-ur-matory agent is evening primrose oil.
The anti-inflammatory agent of the present invention may be used in many distinct physical forms well known in the pharmaceutical art to provide an initial dosage of the anti-inflammatory agent and/or a further time-release form of the anti- inflammatory agent. Without being limited thereto, such physical forms include free forms and encapsulated forms, and mixtures thereof.
The amount of anti-inflammatory agent used in die present invention is a therapeutically effective amount and may vary depending upon the therapeutic dosage recommended or permitted for the particular anti-inflammatory agent. In general, the amount of anti-inflammatory agent present is the ordinary dosage required to obtain the desired result. Such dosages are known to the skilled practitioner in the medical arts and are not a part of the present invention. In a prefeπed embodiment, the anti- inflammatory agent in the sunscreen-wound healing composition is present in an amount from about 0.01% to about 10%, preferably from about 0.1% to about 5%, and more preferably from about 1% to about 3%, by weight.
In yet another preferred embodiment, the therapeutic sunscreen-wound healing compositions of the present invention further comprise a topical anesthetic agent. Anesthetic agents are compounds that induce the loss of tactile sensibility and die sensation of pain. The anesthetic agents in the sunscreen-wound healing compositions of the present invention may be selected from a wide range of therapeutic agents and mixtures of therapeutic agents. Nonlimiting illustrative specific examples of topical anesthetic agents include pramoxine hydrochloride, lidocaine, and benzocaine.
The amount of anesthetic agent used in the present invention is a therapeutically effective amount and may vary depending upon die tiierapeutic dosage recommended or permitted for the particular anesthetic agents. In general, the amount of anesthetic agents present is the ordinary dosage required to obtain the desired result. Such dosages are known to the skilled practitioner in the medical arts and are not a part of the present invention. In a prefeπed embodiment, the anesthetic agent in the sunscreen-wound healing composition is present in an amount from about 1 % to about 30%, preferably from about 2% to about 25%, and more preferably from about 2.5% to about 20%, by weight. B. Methods For Making
The Sunscreen-Wound Healing Compositions
Of Embodiment Two (I.A-D + X)
The present invention extends to methods for making the therapeutic sunscreen-wound healing compositions (LA-D + X). In general, a therapeutic sunscreen-wound healing composition is made by forming an admixture of the wound healing components of Embodiment One (LA-D), a sunscreen agent, and an anti- inflammatory agent. In a first aspect of Embodiment Two (IA + X), a sunscreen- wound healing tiierapeutic composition is made by forming an admixture of a sunscreen agent, an anti-inflammatory agent, and a wound healing composition comprising (a) a pyruvate, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids. In a second aspect of Embodiment Two (I.B + X), a sunscreen-wound healing therapeutic composition is made by forming an admixture of a sunscreen agent, an anti-inflammatory agent, and a wound healing composition comprising (a) a pyruvate, (b) a lactate, and (c) a mixture of saturated and unsaturated fatty acids. In a third aspect of Embodiment Two (LC + X), a sunscreen-wound healing therapeutic composition is made by forming an admixture of a sunscreen agent, an anti-inflammatory agent, and a wound healing composition comprising (a) an antioxidant, and (b) a mixture of saturated and unsaturated fatty acids. In a fourth aspect of Embodiment Two (I.D + X), a sunscreen-wound healing therapeutic composition is made by forming an admixture of admixture of a sunscreen agent, an anti-inflammatory agent, and a wound healing composition comprising (a) a lactate, (b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids.
In a preferred embodiment, die invention is directed to a method for preparing a therapeutic sunscreen-wound healing composition (IA + X) useful to minimize and treat sunburn damage which comprises the steps of admixing a therapeutically effective amount of the following ingredients:
(1) a sunscreen agent;
(2) an anti-inflammatory agent; and (3) a wound healing composition comprising:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are tiiose fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
C. Methods For Employing The Sunscreen-Wound Healing Compositions
Of Embodiment Two (LA-D + X)
The present invention extends to metiiods for employing the therapeutic sunscreen-wound healing compositions (LA-D + X). In general, a therapeutic composition is employed by contacting the therapeutic composition with the skin to be exposed to the sun. In a prefeπed embodiment, die invention is directed to a method for minimizing and treating sunburn in a human with a sunscreen-wound healing composition (LA + X) which comprises the steps of:
(A) providing a tiierapeutically effective amount of a sunscreen-wound healing composition which comprises:
(1) a sunscreen agent;
(2) an anti-inflammatory agent; and
(3) a wound healing composition comprising:
(a) pyruvate selected from die group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and (B) contacting the sunscreen-wound healing composition with the skin of the human prior to exposure to the sun. D. Augmented Sunscreen-Wound Healing Compositions Of Embodiment Two (I.A-D + X + M)
In another aspect of Embodiment Two, the therapeutic sunscreen- wound healing compositions (LA-D + X) of the present invention may be further combined with medicaments useful for treating wounds (M) to form augmented sunscreen-wound healing compositions (I.A-D + X + M). In this embodiment, die combination of the sunscreen-wound healing composition of the present invention and the medicament useful for treating wounds provides an augmented sunscreen-wound healing composition having an enhanced ability to increase the proliferation and resuscitation rate of mammalian cells. For example, the therapeutic compositions of the present invention may be used in combination with medicaments useful for treating wounds such as immunostimulating agents (Betafectin ), antiviral agents, antikeratolytic agents, anti-inflammatory agents, antifungal agents, tretinoin, other sunscreen agents, dermatological agents, topical antihistamine agents, antibacterial agents, bioadhesive agents, respiratory bursting inhibitors (lactic acid, adenosine), inhibitors of prostaglandin synthesis (ibuprofen, aspirin, indomethacin, meclofenomic acid, retinoic acid, padimate O, meclomen, oxybenzone), steroidal anti-inflammatory agents (corticosteroids including synthetic analogs), antimicrobial agents (neosporin ointment, silvadine), antiseptic agents, anesthetic agents (pramoxine hydrochloride, lidocaine, benzocaine), cell nutrient media, bu relief medications, sun bum medications, acne preparations, insect bite and sting medications, wound cleansers, wound dressings, scar reducing agents (vitamin E), and the like, and mixtures thereof, to further enhance the proliferation and resuscitation rate of rnammalian cells. Preferably, the medicament useful for treating wounds is selected from the group consisting of immunostimulating agents, antiviral agents, antikeratolytic agents, anti-inflammatory agents, antifungal agents, tretinoin, sunscreen agents, dermatological agents, topical antihistamine agents, antibacterial agents, bioadhesive agents, respiratory bursting inhibitors, inhibitors of prostaglandin synthesis, antimicrobial agents, cell nutrient media, scar reducing agents, and mixtures thereof. More preferably, the medicament useful for treating wounds is selected from die group consisting of immunostimulating agents, antiviral agents, antikeratolytic agents, anti-inflammatory agents, antifungal agents, acne treating agents, sunscreen agents, dermatological agents, antihistamine agents, antibacterial agents, bioadhesive agents, and mixtures thereof.
In a preferred embodiment, the invention is directed to an augmented sunscreen-wound healing composition (IA + X + M) useful to minimize and treat sunburn damage which comprises:
(A) a therapeutic sunscreen-wound healing composition which comprises a therapeutically effective amount of:
(1) a sunscreen agent; (2) an anti-inflammatory agent; and
(3) a wound healing composition, wherein the wound healing composition comprises:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof; (b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein die fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(B) a medicament useful for treating wounds.
The present invention extends to methods for making the augmented sunscreen-wound healing compositions. In general, the augmented compositions are made by admixing the therapeutic sunscreen-wound healing composition with the medicament useful for treating wounds to prepare the augmented sunscreen-wound healing composition.
The present invention also extends to metiiods for employing the augmented sunscreen-wound healing compositions. In general, an augmented sunscreen-wound healing composition is employed by contacting the composition with the skin to be exposed to the sun. In a specific embodiment, the invention is directed to a method for minimizing and treating sunburn in a human with an augmented sunscreen-wound healing composition (LA + X + M) which comprises the steps of: (A) providing a therapeutically effective amount of a sunscreen-wound healing composition which comprises:
(1) a sunscreen agent;
(2) an anti-inflammato-y agent; and (3) a wound healing composition, wherein the wound healing composition comprises:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(B) a medicament useful for treating wounds.
(C) contacting the augmented sunscreen-wound healing composition with the skin of the human prior to exposure to the sun.
The types of wounds which may be healed using the sunscreen-wound healing compositions and the augmented sunscreen-wound healing compositions of the present invention are inflammation wounds induced by sunburn. The therapeutic compositions may be used topically to protect and accelerate the healing of injured tissue.
Methods for treating sunburn comprise topically administering the compositions of the present invention directly to the skin of the human prior to exposure to the sun. The composition is maintained in contact with the skin for a period of time sufficient to increase the proliferation and resuscitation rate of the cells. E. Formulations Of
The Sunscreen-Wound Healing Compositions
Of Embodiment Two (LA-D + X) and (LA-D + X +M)
Once prepared, die inventive therapeutic sunscreen-wound healing compositions and augmented sunscreen-wound healing compositions may be stored for future use or may be formulated in effective amounts with pharmaceutically acceptable carriers such as pharmaceutical appliances and topical vehicles to prepare a wide variety of pharmaceutical compositions. The pharmaceutically acceptable carriers which may be employed and die metiiods used to prepare the pharmaceutical compositions have been described above in connection witii the formulations of the wound healing compositions of Embodiment One (LA-D).
In a preferred embodiment, die invention is directed to a sunscreen- wound healing pharmaceutical composition which comprises:
(A) a therapeutic sunscreen-wound healing composition (LA + X) which comprises:
(1) a sunscreen agent;
(2) an anti-inflammatory agent; and
(3) a wound healing composition, wherein the wound healing composition comprises:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(B) a pharmaceutically acceptable carrier selected from the group consisting of pharmaceutical appliances, bioadhesives, and occlusive vehicles.
In another prefeπed embodiment, the invention is directed to a method for preparing a pharmaceutical composition for increasing the proliferation and resuscitation rate of mammalian cells, which comprises the steps of: (A) providing a therapeutically effective amount of a sunscreen-wound healing composition (LA + X) which comprises:
(1) a sunscreen agent;
(2) an anti-inflammatory agent; and (3) a wound healing composition comprising:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are tiiose fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells;
(B) providing a pharmaceutically acceptable carrier; and
(C) admixing the sunscreen-wound healing composition from step (A) and die pharmaceutically acceptable carrier from step (B) to form a pharmaceutical composition.
F. Examples Of The Sunscreen-Wound Healing Compositions Of Embodiment Two (LA-D + X) Study 1
SUMMARY
This study was conducted to determine the potential of formulations with moisturizers and antioxidants to effectively prevent or reduce solar simulator derived erythema in normal volunteers, and whether the erythema once manifested would be ameliorated with the test product. The concept was to develop a product tiiat could protect and heal skin from UV damage without the use of UV sunscreen blockers, i.e., PABA, oxybenzone, etc. Such protection would be extremely beneficial for cold sores and psoriatic lesions since UV exposure is known to exacerbate these skin diseases. Two products, a lotion and a cream, were evaluated for their sun protection potential (Phase I) and for product efficacy in the treatment of solar simulator induced erythema (Phase II). Ten subjects completed Phase I of the study. Thirteen subjects completed Phase II of the study. An 8% Homosalate solution served as a control in Phase I.
Under the conditions employed in this study, both the cream and lotion test products provided low level sun protection (SPF values less then 2) and minor improvements in UV-induced erythema as compared witii iπadiated control sites not treated with test products.
INTRODUCTION
Objective
This study was conducted to determine the potential for two wound healing compositions containing formulations for preventing solar simulator derived erythema in normal volunteers and whether, the erythema once manifested, would be ameliorated witii the test wound healing composition product.
Rationale
The demands for sun screen and skin care products make up the fastest growing cosmetics sector in the United States. The growth in sun care products is continuing because overexposure to the sun is believed to produce many ill-effects on the skin. It is known that even high sun protectant factor (SPF- 15) products do not fully protect human skin against cumulative skin damage. The nature of this damage is believed to be associated witii the presence of various "reactive" oxygen species. A product to reduce die levels of photogenerated reactive oxygen species should provide meaningful long term skin benefits. The purpose of this experiment was to determine whether a formula which contains moisturizers and antioxidants could effectively eliminate or reduce the damage produced from overexposure to UV light. The objective was also to determine if applying the product before and/or after UV damage would provide beneficial effects to the damaged area.
Background
Two (2) products were submitted for testing in this two phase study.
Each study group consisted of ten (10) volunteer subjects. The products were considered reasonably safe for testing on human subjects. The protocol followed for Phase I was that established by die OTC panel and published in the Federal Register,
August 25, 1978 (Vol 43, No. 166).
MATERIALS AND METHODS
A sample of each test material(s) was reserved and stored for a period of five (5) years. At the conclusion of the clinical study, the remaining test material(s) was discarded. All information regarding the receipt, storage and disposition of the material(s) was also recorded on a Clinical Material Record form. All test materials were kept in a locked product storage room accessible to clinical staff members only.
Test Materials
Product (1) Identification: Lotion
Description: White Lotion Quantity Provided: 1 x 6oz. Amount applied: 0.1g/50cm
Product (2) Identification: Cream
Description: Yellow Cream Quantity Provided: 1 x 4oz.
Amount applied: 0.1g/50cπr Product (1) was Lubriderm™ Lotion containing 2% sodium pyruvate, 1% vitamin E, and 1% chicken fat. Product (2) was Lubriderm™ Cream containing aquaphor/petrolatum cream with 10% sodium pyruvate, 5% vitamin E, and 5% chicken fat.
EXPERIMENTAL DESIGN
Inclusion Data
Individuals included in the study were eighteen (18) years of age or older; were fair and had uniformly-colored skin on the lower thoracic area of the back which would allow a discernible erythema; were free of any systemic or dermatologic disorder which would interfere with the test results; had completed a phototesting Medical Screening form and a Medical/Personal History form; and had read, understood, and signed an informed consent agreement.
Exclusion Data
Individuals excluded in die study were individuals who had any visible skin disease at the test site which would have interfered with the test results; were receiving systemic or topical drugs or medication such as antihistamines or anti-inflammatories which might have interfered with the test results; were taking medication suspected of causing photobiological reactions (i. e., tetracyclins, thiazides, etc.); had active atopic dermatitis or eczema; had psoriasis; were currently under treatment for asthma; had cataracts; had a history of skin cancer; were pregnant or planning a pregnancy or nursing a child; or has a known sensitivity to cosmetics, skin care products, or topical drugs as related to die products being tested. Light Source
A Xenon Arc Solar Simulator (150W) was used which has a continuous emission spectrum in the UVA and UVB range (290 to 400 nanometers). Less than 1 % of its total energy was composed of "non-solar" wavelengths below 290 nm. The output was monitored daily, using the Robert-Berge meter. Additional measurements of the energy output were taken and recorded as needed.
Definitions of Terms in Clinical Evaluation
MED. Minimal Erythema Dose - The time of light exposure necessary to produce a minimal perceptible erythema (redness) on the skin, discernible sixteen (16) to twenty four (24) hours later.
SPF, Sun Protection Factor - The ultraviolet energy required to produce an MED on protected skin, divided by die ultraviolet energy required to produce an MED on unprotected skin.
PCD, Product Category Designation
1. Minimal Sun Protection Product - Sunscreen products tiiat provide an SPF value of 2 to 4, and offer the least protection from sunbuming, but permit suntanning.
2. Moderate Sun Protection Product - Sunscreen products tiiat provide an SPF value of 4 to 6, and offer moderate protection from sunbuming, and permit some suntanning.
3. Extra Sun Protection Product - Sunscreen products that provide an SPF value of 6 to 8, and offer extra protection from sunbuming, and permit limited suntanning.
4. Maximal Sun Protection Product - Sunscreen products that provide an SPF value of 8 to under 15, and offer maximal protection from sunbuming, and permit little or no suntanning. 5. Ultra Sun Protection Product - Sunscreen products tiiat provide an SPF value of 15 or greater, and offer the most protection from sunbuming and permit no suntanning.
Phase I - Determination of Minimal Erythemal Dose (MED)
MED is defined as die time of light exposure required to produce a minimal perceptible erythema reaction discernible sixteen (16) to twenty-four (24) hours after irradiation using a standardized ultraviolet light source that emits UVB(290- 320 nm) as all or part of its emission spectrum. Subjects who were candidates for this testing have skin types commonly referred to as Fitzpatrick skin types, of Category I, EL, or in according to the following definitions:
Category I Always bu s easily, never tans. Category II Always bums easily, tans minimally.
Category HI Burns moderately, tans gradually (light brown).
Category IV Bums minimally, always tans well (moderate brown).
Category V Rarely burns, tans very well (moderate brown).
Category VI Never bums, deeply pigmented.
An area, other than the test site, was drawn with a marking pen on the subject's back and divided into five (5) equal sites. The anticipated MED was estimated from skin type (I, π, or HI) of the individual and die irradiation size calculated based on energy output of the Xenon lamp. The (5) sites were irradiated on the basis of a geometric progression of 1.25, i.e., each site receiving 25% more exposure than the site to its left. Sixteen (16) to twenty-four (24) hours later, the MED was determined by establishing the site which exhibited die least amount of perceptible erythema. SPF Determination
The test material was applied as follows:
A 50 cm area was marked on the back of the subject. Then, a uniform application of 0.1 g or ml or product was applied to die area with careful spreading and gende but thorough rubbing over the entire marked area. The product was permitted to dry for fifteen (15) to thirty (30) minutes.
The MED previously determined on untreated skin was multiplied by the expected SPF of die product. For example, for an MED of twenty-five (25) seconds, and a test product whose SPF is estimated at two (2), the median exposure time for this product would be 2 x 25 seconds = 50 seconds.
The irradiation for site exposure was again calculated on the basis of a geometric progression of 1.25 with the site to the right receiving more irradiation tiian the site to its left. The sites were then exposed to ultraviolet light from the solar simulator. The procedure was repeated using the control product, an 8% homosalate standard.
In addition, an area of untreated skin was exposed to ultraviolet light from the solar simulator to again determine the MED of unprotected skin. Sixteen (16) to twenty-four (24) hours later, the MED for the treated and untreated skin was determined and the SPF calculated.
Following exposure, all immediate responses were noted for reddening, vesiculation, tanning, darkening of skin, etc. Study Flow Chart Day
1 Obtained informed consent; completed medical screening form; conducted UV irradiation for MED determination.
2 Determined MED; calculated UV exposure times; irradiated test product site, control product site, and untreated sites for second MED determination.
3 Evaluated all sites; calculated SPF.
To assure the uniform evaluation of sunscreen products, a standard sunscreen was used concomitantiy in the test procedure, specifically in step 2, irradiated control product site. This control product was an eight percent (8%) homosalate preparation.
Rejection of Test Data
There were three (3) reasons for rejection of test data.
1. Sometimes the exposure either failed to elicit an MED response on the treated or unprotected skin sites or elicited responses at all five (5) irradiated sites. In either event, that test was considered a technical failure and had to be discarded. If die subject reacted to one or more exposures on the unprotected control site, but not on the treated site, then a minimal estimate could be obtained. However, this estimate would not be used in assessing the Mean of the SPF values.
2. The response on the treated sites was randomly absent, which indicated die product was not spread evenly. Therefore, no assessment of protection was possible. 3. If the SPF values obtained experimentally were well outside die expected range, the values were discarded.
Phase H - Treatment of Erythema
Ten subjects whose MEDs were determined by die previous method (Phase I) had twelve test site areas delineated on the back: five sites were exposed to one times the MED; five sites were exposed to two times the MED, and two sites were not exposed to UV light. The cream test product was applied (0.1g/50cm ) and rubbed into two of the 1 x MED sites, two of the 2 x MED sites, and to one of die unexposed sites. The lotion test product was applied to two separate 1 x MED sites, two separate 1 x MED sites, and one separate unexposed site. The two remaining exposed areas (one 1 x MED and one 2 x MED) served as controls and did not have product applied. All sites were covered, but not occluded, witii a gauze dressing. Application of die product to the same sites was repeated approximately six hours later and the following mornings and afternoons for four consecutive days. Evaluation of the test sites was made each afternoon prior to product application by a trained clinical evaluator who had no participation in any other aspect of the study procedure. A final evaluation was made on die morning of Day 8, the last product application having occurred on die previous Friday (Day 4). Evaluations of each test site were made according to die following scale:
0.0 - minimal or doubtful erythema 0.5 - faint erythema
1.0 - distinct, but mild erythema 2.0 - moderate erythema Results and Discussion
Phase I
Two (2) products, a lotion and a cream, were submitted for evaluation of their sun protective potential. A total of ten (10) subjects between the ages of 25 and 54 were enrolled and completed die evaluation of the test products and die homosalate control.
A low level of sun protection was observed; an SPF value of 1.9 ± 0.2 (N=9) was obtained for the cream and a value of 1.7 ± 0.3 (N=8) was obtained for the lotion.
Phase π
The same two (2) products were also evaluated for their product efficacy in the treatment of solar-simulator induced erythema. A total of fifteen (15) subjects between the ages of 20 and 66 were enrolled and tiiirteen (13) subjects completed die study. Two (2) subjects discontinued for personal reasons.
Product treated sites exposed to two MEDs of irradiation exhibited minor improvements in resolution of erythema as compared to die iπadiated non-product treated sites. Comparisons between erythema levels at Day 2 and subsequent evaluation days was used as die basis of analysis. Day 2 was selected as the baseline day since increases in erythema from Day 1 to Day 2 occurred frequently but appeared to have stabilized by Day 2. Comparison of erythema results on Days 3, 4, 5 and 8 to Day 2 suggest a minor acceleration in the resolution of erythema responses. Specifically, in 12 instances among the 52 comparisons (23%), thirteen (13) subjects with 4 comparisons each, comparing Day 2 to Days 3, 4, 5, and 8, reductions in erythema associated witii both the cream and die lotion were manifested. While it is noted with caution that both the proportion and degree of improvement associated with the product test sites were minor, it is also noteworthy that there was only one instance out of the total of one hundred four (104) comparisons for the two products in which the control site showed greater degree of improvement tiian the test site. Furthermore, this one improvement occurred only for the Day 8/Day
2 comparison.
The cream product showed improvement over the control site in two subjects for all four comparison intervals, in one subject at two of the four intervals, and two other subjects at one of the four intervals. Similarly, the lotion product showed improvement in one subject at all four intervals, in one subject at three intervals, in two subjects at two intervals, and, in one subject at one interval. Conversely, eight of thirteen subjects showed no differences at any of die four intervals in comparisons between the test product sites and the control sites for both the cream and die lotion products. While the effects noted here must again be stressed as minimal, it is, nonetheless, noteworthy that improvements, where noted, favored effects of the test products over the control and not the opposite: twenty-four instances of possible positive effects from the two test products compared to one such possible instance at the control site.
A similar analysis of data associated with the 1 x MED sites did not provide the same results due to the low level and variable erythematous responses observed. This is believed to be due to the fact that minor perturbations of ultra violet lamp output are likely to be sufficient to result in sub-erythema and mildly erythema responses even on the same subject at adjoining sites.
In conclusion, under die conditions employed in this clinical study, both the cream and lotion test products provided low-level sun protection and indications of minor improvement in UV -induced erythema as compared with iπadiated control sites not treated witii test products . However, it is clear that these effects are, at best, minor and that verification of efficacy would require a larger proportion of study subjects. Formulation changes (vehicle, level of wound healing composition) or product application (e.g., multiple applications or occlusive conditions) may also be worth exploring.

Claims

I claim:
1. A therapeutic sunscreen-wound healing composition useful to minimize and treat sunburn damage which comprises a tiierapeutically effective amount of:
(1) a sunscreen agent;
(2) an anti-inflammatory agent; and
(3) a wound healing composition, wherein the wound healing composition comprises:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
2. The composition according to claim 1, wherein the sunscreen agent is selected from the group consisting of ethylhexyl p-methoxycinnamate, octyl methoxycinnamate, octyl dimethyl /?-aminobenzoic acid, 2-ethylhexyl salicylate, octyl salicylate, menthyl anthranilate, octocrylene, padimate o, titanium dioxide, urea, and oxybenzone.
3. The composition according to claim 2, wherein the sunscreen agent is oxybenzone.
4. The composition according to claim 1 , wherein the anti-inflammatory agent is selected from the group consisting of ibuprofen, naproxen, sulindac, diflunisal, piroxicam, indomethacin, etodolac, meclofenamate sodium, fenoproben calcium, ketoprofen, mefenamic acid, nabumetone, ketorolac tromethamine, diclofenac, evening primrose oil, acetylsalicylic acid, mesalamine, salsalate, diflunisal, salicylsalicylic acid, choline magnesium trisalicylate, flunisolide, triamcinoline, triamcinoline acetonide, beclometiiasone diproprionate, betamethasone diproprionate, hydrocortisone, cortisone, dexamethasone, predinisone, methyl prednisolone, and prednisolone.
5. The composition according to claim 4, wherein the anti-inflammatory agent is evening primrose oil.
6. The composition according to claim 1, wherein the pyruvate is selected from the group consisting of pyruvic acid, litiiium pymvate, sodium pymvate, potassium pymvate, magnesium pymvate, calcium pymvate, zinc pymvate, manganese pymvate, methyl pymvate, σ-ketoglutaric acid, pharmaceutically acceptable salts of pymvic acid, prodrugs of pymvic acid, and mixtures thereof.
7. The composition according to claim 6, wherein the pymvate is sodium pymvate.
8. The composition according to claim 1, wherein the antioxidant is selected from the group consisting of all forms of Vitamin A including retinol and 3,4- didehydoretinol, all forms of carotene including α-carotene, ^-carotene, gamma- carotene, and -te/ta-carotene, all forms of Vitamin C including D-ascorbic acid and L- ascorbic acid, all forms of Vitamin E including -tocopherol, yS- tocopherol, gamma- tocopherol, -fe/tα-tocopherol, tocoquinone, tocotrienol, Vitamin E esters which readily undergo hydrolysis to Vitamin E including Vitamin E acetate and Vitamin E succinate, and pharmaceutically acceptable Vitamin E salts such as Vitamin E phosphate, prodrugs of Vitamin A, carotene, Vitamin C, and Vitamin E, pharmaceutically acceptable salts of Vitamin A, carotene, Vitamin C, and Vitamin E, and mixtures thereof.
9. The composition according to claim 8, wherein the antioxidant is Vitamin E acetate.
10. The composition according to claim 1, wherein the mixture of saturated and unsaturated fatty acids is selected from the group consisting of animal and vegetable fats and waxes.
11. The composition according to claim 10, wherein the mixture of saturated and unsaturated fatty acids is selected from the group consisting of human fat, chicken fat, cow fat, sheep fat, horse fat, pig fat, and whale fat.
12. The composition according to claim 11, wherein die mixture of saturated and unsaturated fatty acids comprises lauric acid, myristic acid, myristoleic acid, pentadecanoic acid, palmitic acid, paimitoleic acid, margaric acid, margaroleic acid, stearic, oleic acid, linoleic acid, linolenic acid, arachidic acid, and gadoleic acid.
13. The composition according to claim 1, wherein die sunscreen agent is present in the therapeutic wound healing composition in an amount from about 1% to about 30%, by weight of the therapeutic wound healing composition.
14. The composition according to claim 1, wherein the anti- inflammatory agent is present in the therapeutic wound healing composition in an amount from about 0.01% to about 10%, by weight of the therapeutic wound healing composition.
15. The composition according to claim 1, wherein pymvate is present in the therapeutic wound healing composition in an amount from about 10% to about
50%, by weight of the therapeutic wound healing composition.
16. The composition according to claim 1, wherein the antioxidant is present in the therapeutic wound healing composition in an amount from about 0.1% to about 40%, by weight of the therapeutic wound healing composition.
17. The composition according to claim 1, wherein the mixture of saturated and unsaturated fatty acids is present in the therapeutic wound healing composition in an amount from about 10% to about 50%, by weight of the therapeutic wound healing composition.
18. The composition according to claim 1, further comprising a therapeutically effective amount of a topical anesthetic.
19. A method for minimizing and treating sunburn in a human with a sunscreen-wound healing composition which comprises the steps of:
(A) providing a therapeutically effective amount of a sunscreen-wound healing composition which comprises:
(1) a sunscreen agent;
(2) an anti-inflammatory agent; and (3) a wound healing composition comprising:
(a) pymvate selected from the group consisting of pymvic acid, pharmaceutically acceptable salts of pymvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(B) contacting the sunscreen-wound healing composition with the skin of the human prior to exposure to the sun.
20. The method according to claim 19, further comprising a therapeutically effective amount of a topical anesthetic.
21. A method for preparing a therapeutic sunscreen-wound healing composition useful to minimize and treat sunburn damage which comprises the steps of admixing a tiierapeutically effective amount of the following ingredients:
(1) a sunscreen agent;
(2) an anti-inflammatory agent; and (3) a wound healing composition comprising:
(a) pymvate selected from the group consisting of pymvic acid, pharmaceutically acceptable salts of pymvic acid, and mixtures thereof;
(b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.
22. The method according to claim 21, further comprising a therapeutically effective amount of a topical anesthetic.
23. An augmented sunscreen-wound healing composition useful to minimize and treat sunburn damage which comprises:
(A) a therapeutic sunscreen-wound healing composition which comprises a therapeutically effective amount of:
(1) a sunscreen agent;
(2) an anti-inflammatory agent; and
(3) a wound healing composition, wherein the wound healing composition comprises: (a) pymvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pymvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are tiiose fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(B) a medicament useful for treating wounds.
24. The composition according to claim 23, further comprising a therapeutically effective amount of a topical anesthetic.
25. The augmented antiacne- wound healing composition according to claim 23, wherein the medicament useful for treating wounds is selected from the group consisting of immunostimulating agents, antiviral agents, antikeratolytic agents, anti-inflammatory agents, antifungal agents, acne treating agents, other sunscreen agents, dermatological agents, antihistamine agents, antibacterial agents, bioadhesive agents, respiratory bursting inhibitors, inhibitors of prostaglandin synthesis, antimicrobial agents, antiseptic agents, anesthetic agents, cell nutrient media, bum relief medications, sun bum medications, insect bite and sting medications, wound cleansers, wound dressings, scar reducing agents, and mixtures thereof.
26. A method for minimizing and treating sunburn in a human with an augmented sunscreen-wound healing composition which comprises the steps of:
(A) providing a therapeutically effective amount of a sunscreen-wound healing composition which comprises:
(1) a sunscreen agent;
(2) an anti-inflammatory agent; and (3) a wound healing composition, wherein the wound healing composition comprises:
(a) pymvate selected from the group consisting of pymvic acid, pharmaceutically acceptable salts of pymvic acid, and mixtures thereof;
(b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(B) a medicament useful for treating wounds.
(C) contacting the augmented sunscreen-wound healing composition with the skin of the human prior to exposure to the sun.
27. The method according to claim 26, further comprising a therapeutically effective amount of a topical anesthetic.
28. A sunscreen-wound healing pharmaceutical composition which comprises:
(A) a therapeutic sunscreen-wound healing composition which comprises: (1) a sunscreen agent;
(2) an anti-inflammatory agent; and
(3) a wound healing composition, wherein the wound healing composition comprises: (a) pymvate selected from the group consisting of pymvic acid, pharmaceutically acceptable salts of pymvic acid, and mixtures thereof;
(b) an antioxidant; and
(c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are tiiose fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and
(B) a pharmaceutically acceptable carrier selected from the group consisting of pharmaceutical appliances, bioadhesives, and occlusive vehicles.
29. The composition according to claim 28, further comprising a therapeutically effective amount of a topical anesthetic.
PCT/US1995/012848 1994-12-07 1995-10-05 Sunscreen-wound healing composition WO1996017624A1 (en)

Priority Applications (5)

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AU38596/95A AU690366B2 (en) 1994-12-07 1995-10-05 Sunscreen-wound healing composition
NZ295303A NZ295303A (en) 1994-12-07 1995-10-05 Sunscreen composition comprising a sunscreen agent, an anti-inflammatory agent and a composition for healing wounds
MX9703556A MX9703556A (en) 1994-12-07 1995-10-05 Sunscreen-wound healing composition.
EP95936858A EP0796107B1 (en) 1994-12-07 1995-10-05 Sunscreen-wound healing composition
DE69529345T DE69529345T2 (en) 1994-12-07 1995-10-05 Wound-healing sunscreen

Applications Claiming Priority (4)

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US35091894A 1994-12-07 1994-12-07
US08/350,918 1994-12-07
US08/446,979 US5674912A (en) 1991-03-01 1995-05-22 Sunscreen-wound healing compositions and methods for preparing and using same
US08/446,979 1995-05-22

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EP (1) EP0796107B1 (en)
AU (1) AU690366B2 (en)
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MX (1) MX9703556A (en)
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0901368A1 (en) * 1996-02-19 1999-03-17 Monash University Dermal penetration enhancers and drug delivery systems involving same
WO2002062744A1 (en) * 2001-02-06 2002-08-15 The Nisshin Oillio, Ltd. Utilization for improving stability of uv-absorbing substances, uv-absorbers and cosmetics
EP0784975B1 (en) * 1995-12-26 2005-10-19 Teikoku Seiyaku Kabushiki Kaisha Use of acetylsalicylic acid in the manufacture of a drug for the treatment of skin injuries
WO2007026240A1 (en) * 2005-09-01 2007-03-08 Medical Therapies Limited Sunscreen and cosmetic compositions for prophylaxis or treatment of skin cancers
DE10196483B4 (en) 2000-09-01 2023-08-24 GSK Consumer Healthcare S.A. treatment of sunburn

Families Citing this family (114)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6703009B1 (en) 1993-03-30 2004-03-09 Tendskin International Inc. Topical compositions and methods for treating pseudofolliculitis barbae and ingrown hair
JP3971454B2 (en) * 1993-10-29 2007-09-05 ザ トラスティーズ オブ ボストン ユニバーシティ Physiologically stable compositions of butyric acid, butyrate, and derivatives as anti-neoplastic agents
IT1276698B1 (en) * 1995-06-09 1997-11-03 Sa Fo Sa Srl NATURAL BASED SCREENING AND ANTIERITEMA MIXTURE
US5885593A (en) * 1995-09-28 1999-03-23 The Andrew Jergens Company Skin care composition including cyclodextrin materials and method for treating skin therewith
AUPO379596A0 (en) 1996-11-22 1996-12-19 Soltec Research Pty Ltd Percutaneous delivery system
WO1999040883A2 (en) * 1998-02-11 1999-08-19 Faller Douglas V Compositions and methods for the treatment of cystic fibrosis
US5980920A (en) * 1998-03-02 1999-11-09 Lindquist; Niels L. Antioxidant compositions
AUPP229598A0 (en) * 1998-03-11 1998-04-09 Oldfield Family Holdings Pty Limited Treatment of sunburn
US6372712B1 (en) 1998-05-22 2002-04-16 The Board Of Trustees Of The Leland Stanford Jr. University Synthetic bifunctional molecules containing a drug moiety and presenter protein ligand
WO1999066797A1 (en) 1998-06-22 1999-12-29 Worden Charles E Enriched platelet wound healant
AU768543B2 (en) * 1998-06-22 2003-12-18 Nuo Therapeutics, Inc. Improved enriched platelet wound healant
DE19830770C1 (en) * 1998-07-09 2000-05-18 Sueddeutsche Kalkstickstoff Water-soluble zinc pyruvates and their hydrates, process for their preparation and their use
US6503488B1 (en) * 1998-11-17 2003-01-07 Tend Skin International, Inc. Topical compositions including deodorant compositions
US6469066B1 (en) * 1998-12-18 2002-10-22 Palladin Healthcare International, Ltd. Composition for pain mediation and apparatus and method of use thereof
US6627178B1 (en) * 1999-07-30 2003-09-30 Garret D. Cawthon Methods, compositions and systems for the prevention and treatment of diaper rash
US6667045B2 (en) 1999-10-01 2003-12-23 Joseph Scott Dahle Topical applications for skin treatment
ATE285223T1 (en) * 1999-10-08 2005-01-15 Coty Bv COSMETIC ACTIVE INGREDIENT PREPARATION WITH SYNERGISTICALLY INCREASED RADICAL PROTECTION FACTOR
EP1642885B1 (en) 2000-08-29 2009-11-11 Biocon Limited Use of a pharmaceutical composition containing a para-aminophenyl acetic acid derivative for treating inflammatory conditions of the gastrointestinal tract
US20020082222A1 (en) * 2000-11-30 2002-06-27 Shapira Nathan Andrew Treatments for neurogenetic disorders, impulse control disorder, and wound healing
GB0030460D0 (en) * 2000-12-14 2001-01-24 Isis Innovation Drug delivery system
DE10100127A1 (en) * 2001-01-03 2002-10-02 Henkel Kgaa Procedure for determining the homeostasis of the skin
AU2002254525B2 (en) * 2001-04-04 2004-12-23 Critical Therapeutics, Inc. Method for preventing acute renal failure
US7238680B2 (en) * 2001-06-01 2007-07-03 Rosen Steven E Topical compositions for veterinary uses
ATE445838T1 (en) 2001-07-25 2009-10-15 Raptor Pharmaceutical Inc COMPOSITIONS AND METHODS FOR MODULATING TRANSPORT ACROSS THE BLOOD-BRAIN BARRIER
US6756059B2 (en) * 2001-08-20 2004-06-29 Skinvisible Pharmaceuticals, Inc. Topical composition, topical composition precursor, and methods for manufacturing and using
US8048924B2 (en) 2001-08-29 2011-11-01 Biocon Limited Methods and compositions employing 4-aminophenylacetic acid compounds
US8076373B2 (en) * 2001-09-11 2011-12-13 North Cell Pharmacetical Method for treating mammalian diseases and injuries caused by the over-expression of peroxynitrite
US7037513B1 (en) 2005-01-31 2006-05-02 Aquea Scientific Corporation Bodywash additives
US7025952B1 (en) 2005-01-31 2006-04-11 Aquea Scientific Corporation Methods of preparation and use of bodywashes containing additives
US6998113B1 (en) * 2005-01-31 2006-02-14 Aquea Scientific Corporation Bodywashes containing additives
US20060173709A1 (en) * 2005-01-31 2006-08-03 Traynor Daniel H Bodywash additive business methods
US20030143165A1 (en) * 2002-01-25 2003-07-31 Allan Evans NSAID-containing topical formulations that demonstrate chemopreventive activity
JP2005526832A (en) * 2002-04-17 2005-09-08 クリティカル セラピューティクス，インコーポレイテッド Pharmaceutical composition containing α-ketoalkanoic acid ester or -amide and lactic acid or lactate
US20040018237A1 (en) * 2002-05-31 2004-01-29 Perricone Nicholas V. Topical drug delivery using phosphatidylcholine
US8435942B2 (en) * 2002-05-31 2013-05-07 Transdermal Biotechnology, Inc. Methods for formulating stabilized insulin compositions
DE10226990A1 (en) * 2002-06-18 2004-03-18 Sanguibiotech Ag Topically applicable micro-emulsions with binary phase and active substance differentiation, their production and their use, in particular for supplying the skin with bioavailable oxygen
KR20120008081A (en) * 2002-10-25 2012-01-25 레반스 테라퓨틱스, 아이엔씨. Modulation of zinc levels to improve tissue properties
US20040186082A1 (en) * 2003-03-20 2004-09-23 Hartman Raymond A. Enhanced phototherapy for the treatment of cancer and autoimmune disease
US7714015B2 (en) * 2003-08-07 2010-05-11 Lil Brat Pharmaceuticals Of Marlette, Mi Method and composition for treating sunburned skin
CN1886128A (en) * 2003-09-30 2006-12-27 柯西有限公司 Compositions and methods for treating burns
DK1773767T3 (en) 2004-07-07 2016-03-21 Biocon Ltd Synthesis of azo bound in immune regulatory relations
US20100266709A1 (en) * 2004-12-16 2010-10-21 Hicks Terry Lee Compositions and Methods for Treating Burns
US7001592B1 (en) 2005-01-31 2006-02-21 Aquea Scientific Corporation Sunscreen compositions and methods of use
US20080112904A1 (en) * 2005-03-08 2008-05-15 Daniel Henry Traynor Sunscreen Compositions And Methods Of Use
CA2789262C (en) 2005-04-28 2016-10-04 Proteus Digital Health, Inc. Pharma-informatics system
US20070048233A1 (en) * 2005-08-26 2007-03-01 Spray Tanning, Inc. Artificial tanning composition containing medication
US20070048393A1 (en) * 2005-09-01 2007-03-01 Spray Tanning, Inc. Topical turmeric skin care products
US7763289B2 (en) * 2005-09-01 2010-07-27 JoAl's Products, LLC Topical turmeric skin care products
FR2896117A1 (en) * 2006-01-06 2007-07-13 France Telecom METHODS OF ENCODING AND DECODING AN IMAGE SEQUENCE, DEVICES, COMPUTER PROGRAMS, AND CORRESPONDING SIGNAL
US7252816B1 (en) 2006-03-29 2007-08-07 Dow Pharmaceutical Sciences Topical acne vulgairs medication with a sunscreen
US8604901B2 (en) * 2006-06-27 2013-12-10 Eyelock, Inc. Ensuring the provenance of passengers at a transportation facility
US20080057138A1 (en) * 2006-09-06 2008-03-06 Telford Holdings Ltd. Restorative skin cream
MX2009002893A (en) 2006-09-18 2009-07-10 Raptor Pharmaceutical Inc Treatment of liver disorders by administration of receptor-associated protein (rap)-conjugates.
WO2008049020A2 (en) 2006-10-17 2008-04-24 Nuvo Research Diclofenac gel
WO2008066618A2 (en) * 2006-10-20 2008-06-05 Skinvisible Pharmaceuticals, Inc. Antifungal composition and methods for using
CN101730518A (en) * 2007-05-21 2010-06-09 阿奎耶科技公司 Highly charged microcapsule
US20090041686A1 (en) * 2007-08-06 2009-02-12 Osborne David W Topical acne vulgaris composition with a sunscreen
US8802073B2 (en) * 2007-11-15 2014-08-12 Prelief Inc. Methods and compositions for wound healing
US8636988B2 (en) * 2008-01-31 2014-01-28 Doctor Essentials Composition for treatment of sunburned skin
US20100099766A1 (en) * 2008-10-16 2010-04-22 Novartis Ag Topical NSAID compositions having sensate component
NZ616673A (en) 2009-02-20 2014-08-29 To Bbb Holding B V Glutathione-based drug delivery system
WO2010105112A1 (en) * 2009-03-11 2010-09-16 Hemaquest Pharmaceuticals, Inc. Detection of short-chain fatty acids in biological samples
US8618164B2 (en) 2009-03-31 2013-12-31 Nuvo Research Inc. Treatment of pain with topical diclofenac compounds
IL295075A (en) 2009-05-06 2022-09-01 Laboratory Skin Care Inc Dermal delivery compositions comprising active agent-calcium phosphate particle complexes and methods of using the same
US20110086869A1 (en) 2009-09-24 2011-04-14 The Trustees Of Boston University Methods for treating viral disorders
EP2509418A4 (en) 2009-12-08 2013-03-20 Hemaquest Pharmaceuticals Inc Methods and low dose regimens for treating red blood cell disorders
WO2011113013A2 (en) 2010-03-11 2011-09-15 Hemaquest Pharmaceuticals, Inc. Methods and compositions for treating viral or virally-induced conditions
US20120077778A1 (en) 2010-09-29 2012-03-29 Andrea Bourdelais Ladder-Frame Polyether Conjugates
CA2719512A1 (en) 2010-11-01 2012-05-01 Stiefel Research Australia Pty Ltd Polymeric topical compositions
CN103547258B (en) 2011-03-17 2017-10-20 特兰斯德梅尔生物工艺股份有限公司 Local nitric oxide system and its application method
US8871261B2 (en) 2012-09-19 2014-10-28 Transdermal Biotechnology, Inc. Cancer treatments and compositions for use thereof
US8871258B2 (en) 2012-09-19 2014-10-28 Transdermal Biotechnology, Inc. Treatment and prevention of learning and memory disorders
US8871262B2 (en) 2012-09-19 2014-10-28 Transdermal Biotechnology, Inc. Compositions and methods for treatment of osteoporosis and other indications
US8871259B2 (en) 2012-09-19 2014-10-28 Transdermal Biotechnology, Inc. Techniques and systems for treatment of neuropathic pain and other indications
US8871254B2 (en) 2012-09-19 2014-10-28 Transdermal Biotechnology, Inc. Systems and methods for treatment of acne vulgaris and other conditions with a topical nitric oxide delivery system
US8871255B2 (en) 2012-09-19 2014-10-28 Transdermal Biotechnology, Inc. Treatment of skin and soft tissue infection with nitric oxide
US8871260B2 (en) 2012-09-19 2014-10-28 Transdermal Biotechnology, Inc. Methods and compositions for muscular and neuromuscular diseases
US8871256B2 (en) 2012-09-19 2014-10-28 Transdermal Biotechnology, Inc. Methods and systems for treatment of inflammatory diseases with nitric oxide
US8871257B2 (en) 2012-09-19 2014-10-28 Transdermal Biotechnology, Inc. Prevention and treatment of cardiovascular diseases using systems and methods for transdermal nitric oxide delivery
EP2916814B1 (en) 2012-11-06 2020-02-12 Colabs International Corporation Composition containing a cellulose derived capsule with a sunscreen
US11690793B2 (en) 2012-11-06 2023-07-04 Colabs Int'l Corp. Composition containing a cellulose derived capsule with a sunscreen
US11491088B2 (en) 2012-11-06 2022-11-08 CoLabs International Corporation Compositions containing a capsule with a moisturizing agent
US11724134B2 (en) 2012-11-06 2023-08-15 CoLabs International Corporation Compositions containing a cellulose derived capsule with a sunscreen active agent
US11707421B2 (en) 2012-11-06 2023-07-25 Colabs Int'l Corp. Compositions containing a flexible derived capsule with an active agent
US10322301B2 (en) 2012-11-06 2019-06-18 CoLabs International Corporation Compositions containing a cellulose derived capsule with a sunscreen active agent
US9717679B1 (en) 2012-12-17 2017-08-01 Touchless Designs LLC Methods for the prevention and treatment of animal skin conditions
US9849160B2 (en) 2013-03-13 2017-12-26 Transdermal Biotechnology, Inc. Methods and systems for treating or preventing cancer
US9314423B2 (en) 2013-03-13 2016-04-19 Transdermal Biotechnology, Inc. Hair treatment systems and methods using peptides and other compositions
US9295637B2 (en) 2013-03-13 2016-03-29 Transdermal Biotechnology, Inc. Compositions and methods for affecting mood states
US9724419B2 (en) 2013-03-13 2017-08-08 Transdermal Biotechnology, Inc. Peptide systems and methods for metabolic conditions
US9314417B2 (en) 2013-03-13 2016-04-19 Transdermal Biotechnology, Inc. Treatment of skin, including aging skin, to improve appearance
US9687520B2 (en) 2013-03-13 2017-06-27 Transdermal Biotechnology, Inc. Memory or learning improvement using peptide and other compositions
US9387159B2 (en) 2013-03-13 2016-07-12 Transdermal Biotechnology, Inc. Treatment of skin, including aging skin, to improve appearance
US20140271937A1 (en) 2013-03-13 2014-09-18 Transdermal Biotechnology, Inc. Brain and neural treatments comprising peptides and other compositions
US9295647B2 (en) 2013-03-13 2016-03-29 Transdermal Biotechnology, Inc. Systems and methods for delivery of peptides
US9320758B2 (en) 2013-03-13 2016-04-26 Transdermal Biotechnology, Inc. Brain and neural treatments comprising peptides and other compositions
US9314433B2 (en) 2013-03-13 2016-04-19 Transdermal Biotechnology, Inc. Methods and systems for treating or preventing cancer
US20140271731A1 (en) 2013-03-13 2014-09-18 Transdermal Biotechnology, Inc. Cardiovascular disease treatment and prevention
US9241899B2 (en) 2013-03-13 2016-01-26 Transdermal Biotechnology, Inc. Topical systems and methods for treating sexual dysfunction
US9314422B2 (en) 2013-03-13 2016-04-19 Transdermal Biotechnology, Inc. Peptide systems and methods for metabolic conditions
US9393265B2 (en) 2013-03-13 2016-07-19 Transdermal Biotechnology, Inc. Wound healing using topical systems and methods
US20140271938A1 (en) 2013-03-13 2014-09-18 Transdermal Biotechnology, Inc. Systems and methods for delivery of peptides
US9393264B2 (en) 2013-03-13 2016-07-19 Transdermal Biotechnology, Inc. Immune modulation using peptides and other compositions
US9339457B2 (en) 2013-03-13 2016-05-17 Transdermal Biotechnology, Inc. Cardiovascular disease treatment and prevention
US9320706B2 (en) 2013-03-13 2016-04-26 Transdermal Biotechnology, Inc. Immune modulation using peptides and other compositions
US9295636B2 (en) 2013-03-13 2016-03-29 Transdermal Biotechnology, Inc. Wound healing using topical systems and methods
US9750787B2 (en) 2013-03-13 2017-09-05 Transdermal Biotechnology, Inc. Memory or learning improvement using peptide and other compositions
CA2925539A1 (en) * 2013-10-16 2015-04-23 University Of South Alabama Formulations including silver nanoparticles and methods of using the same
US20150335572A1 (en) * 2014-05-26 2015-11-26 Michael Lee Martin Medicated Hard Candy Product For Treating Esophageal Inflammation And A Method Using The Same
US11135158B2 (en) 2014-05-26 2021-10-05 Michael Lee Martin Medicated hard candy product for treating esophageal inflammation and a method using the same
US20170175080A1 (en) * 2015-12-17 2017-06-22 San Diego Blood Bank Compositions and methods for cell culture
AU2019291885A1 (en) 2018-06-27 2021-02-04 CoLabs International Corporation Compositions comprising silicon dioxide-based particles including one or more agents
AU2020283590A1 (en) 2019-05-31 2022-01-20 Viracta Subsidiary, Inc. Methods of treating virally associated cancers with histone deacetylase inhibitors
CA3074150A1 (en) 2020-02-18 2021-08-18 Ovation Science, Inc. Composition and method for transdermal delivery of cannabidiol (cbd) and delta9-tetrahydrocannabinol (thc)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992015292A1 (en) * 1991-03-01 1992-09-17 Warner-Lambert Company Therapeutic compositions to protect and resuscitate mammalian cells and methods for preparing and using same
WO1993010776A1 (en) * 1991-11-26 1993-06-10 Warner-Lambert Company Wound healing compositions containing a pyruvate, an antioxidant and a mixture of fatty acids

Family Cites Families (40)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3887702A (en) * 1973-08-30 1975-06-03 Mildred Baldwin Composition and method for treating fingernails and toenails
US3879537A (en) * 1973-09-04 1975-04-22 Scott Eugene J Van Treatment of ichthyosiform dermatoses
US4294852A (en) * 1973-11-01 1981-10-13 Johnson & Johnson Skin treating compositions
US3984566A (en) * 1974-02-25 1976-10-05 Scott Eugene J Van Method of alleviating the symptoms of dandruff
US3988470A (en) * 1974-02-25 1976-10-26 Scott Eugene J Van Treatment of palmar and plant disturbed keratosis
US4158057A (en) * 1975-03-28 1979-06-12 Stanko Ronald T Prevention of the accumulation of fatty deposits in the liver
US4197316A (en) * 1975-07-23 1980-04-08 Scott Eugene J Van Treatment of dry skin
US4021572A (en) * 1975-07-23 1977-05-03 Scott Eugene J Van Prophylactic and therapeutic treatment of acne vulgaris utilizing lactamides and quaternary ammonium lactates
US4363815A (en) * 1975-07-23 1982-12-14 Yu Ruey J Alpha hydroxyacids, alpha ketoacids and their use in treating skin conditions
US4170821A (en) * 1977-12-02 1979-10-16 Warner-Lambert Company Razor cartridges
US4284630A (en) * 1978-03-22 1981-08-18 Yu Ruey J Stabilized water-in-oil emulsions
US4234599A (en) * 1978-10-04 1980-11-18 Scott Eugene J Van Treatment of skin keratoses with α-hydroxy acids and related compounds
US4246261A (en) * 1979-08-09 1981-01-20 Scott Eugene J Van Additives enhancing topical corticosteroid action
JPS5783287A (en) * 1980-11-14 1982-05-25 Kyowa Hakko Kogyo Co Ltd Elimination of hydrogen peroxide
US4812479A (en) * 1981-04-01 1989-03-14 The Montefiore Hospital Society Of Western Pennsylvania, Inc. Method for preventing body fat deposition in mammals
US4548937A (en) * 1981-04-01 1985-10-22 Montefiore Hospital Method for preventing body fat deposition in mammals
US4645764A (en) * 1981-04-01 1987-02-24 Montefiore Hospital Method for preventing body fat deposition in animals
US4415576A (en) * 1981-04-01 1983-11-15 Montefiore Hospital Method for preventing body fat deposition in mammals
US4351835A (en) * 1981-04-01 1982-09-28 Montefiore Hospital Method for preventing body fat deposition in mammals
JPS57194787A (en) * 1981-05-28 1982-11-30 Ajinomoto Co Inc Culture medium for animal cell
US4454159A (en) * 1981-12-28 1984-06-12 Albert Musher Dermatological treatment preparations
DE3201511A1 (en) * 1982-01-20 1983-07-28 Henkel Kgaa SEBOSUPPRESSIVE COSMETIC AGENTS WHICH CONTAIN LONG-CHAIN ALKANOLS AND ANTIOXIDANTS
US4451482A (en) * 1982-04-22 1984-05-29 Shell Oil Company Method of preventing or ameliorating pyrethroid skin sensory stimulation
US4521375A (en) * 1982-11-23 1985-06-04 Coopervision, Inc. Sterilizing treatment with hydrogen peroxide and neutralization of residual amounts thereof
WO1985002092A1 (en) * 1983-11-14 1985-05-23 Bio-Mimetics Inc. Bioadhesive compositions and methods of treatment therewith
WO1986000227A1 (en) * 1984-06-22 1986-01-16 Veech Richard L Electrolyte solutions and in vivo use thereof
DE3438455A1 (en) * 1984-10-19 1986-06-26 Institut Dr. Ziegler, Bettingen L-LYSINE AND L-HISTIDINE PYRUVATE
US4696917A (en) * 1985-08-01 1987-09-29 Lindstrom Richard L Irrigation solution
CS262822B1 (en) * 1986-10-03 1989-04-14 Kovar Jan Synthetic medium for the cultivation of myelomic cells
DE3719097C1 (en) * 1987-06-06 1988-06-09 Fratzer Uwe Medicament containing eicosapentaenoic acid and docosahexaenoic acid as unsaturated fatty acids as well as vitamin E.
JPS6432738A (en) * 1987-07-29 1989-02-02 Mitsubishi Electric Corp Synchronism detecting circuit
US4847072A (en) * 1987-10-22 1989-07-11 The Procter & Gamble Company Photoprotection compositions comprising tocopherol sorbate
US4847069A (en) * 1987-10-22 1989-07-11 The Procter & Gamble Company Photoprotection compositions comprising sorbohydroxamic acid and an anti-inflammatory agent
US4847071A (en) * 1987-10-22 1989-07-11 The Procter & Gamble Company Photoprotection compositions comprising tocopherol sorbate and an anti-inflammatory agent
TR22895A (en) * 1988-05-02 1988-10-11 Permatik Celik Ve Plastik San THE PLATFORM AND BASHK SECTIONS ARE OR A LOT OF BICAKH TIRAS URENITIES, WHICH ARE PRODUCED WITH A SUPPLEMENT OF AUXILIARY TIRAS MATERIAL.
KR910000101A (en) * 1988-06-02 1991-01-29 유다까 미시마 Enzyme inhibitor
GB8813766D0 (en) * 1988-06-10 1988-07-13 Efamol Holdings Essential fatty acid compositions
CA2018471A1 (en) * 1989-07-28 1991-01-28 Ian W. Kellaway Mucoadhesive hydrogels delivery system
US5084482A (en) * 1990-04-10 1992-01-28 The Lithox Corporation Methods for inhibiting inflammatory ischemic, thrombotic and cholesterolemic disease response with methionine compounds
US5296307A (en) * 1992-05-08 1994-03-22 Electric Power Research Institute, Inc. Laminated paper polyolefin paper composite

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992015292A1 (en) * 1991-03-01 1992-09-17 Warner-Lambert Company Therapeutic compositions to protect and resuscitate mammalian cells and methods for preparing and using same
WO1993010776A1 (en) * 1991-11-26 1993-06-10 Warner-Lambert Company Wound healing compositions containing a pyruvate, an antioxidant and a mixture of fatty acids

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0784975B1 (en) * 1995-12-26 2005-10-19 Teikoku Seiyaku Kabushiki Kaisha Use of acetylsalicylic acid in the manufacture of a drug for the treatment of skin injuries
EP0901368A1 (en) * 1996-02-19 1999-03-17 Monash University Dermal penetration enhancers and drug delivery systems involving same
EP0901368B1 (en) * 1996-02-19 2006-05-03 Acrux DDS Pty Ltd Dermal penetration enhancers and drug delivery systems involving same
US7387789B2 (en) 1996-02-19 2008-06-17 Acrux Dds Pty. Ltd. Transdermal delivery of non-steroidal anti-inflammatory drugs
US7438203B2 (en) 1996-02-19 2008-10-21 Acrux Dds Pty Ltd Dermal penetration enhancers and drug delivery systems involving same
DE10196483B4 (en) 2000-09-01 2023-08-24 GSK Consumer Healthcare S.A. treatment of sunburn
WO2002062744A1 (en) * 2001-02-06 2002-08-15 The Nisshin Oillio, Ltd. Utilization for improving stability of uv-absorbing substances, uv-absorbers and cosmetics
US7232560B2 (en) 2001-02-06 2007-06-19 The Nisshin Oillio Group, Ltd. Utilization absorber, cosmetic and method for enhancing stability of ultraviolet-absorbing substances
WO2007026240A1 (en) * 2005-09-01 2007-03-08 Medical Therapies Limited Sunscreen and cosmetic compositions for prophylaxis or treatment of skin cancers

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US5674912A (en) 1997-10-07
EP0796107A1 (en) 1997-09-24
DE69529345D1 (en) 2003-02-13
AU3859695A (en) 1996-06-26
AU690366B2 (en) 1998-04-23
EP0796107B1 (en) 2003-01-08
DE69529345T2 (en) 2003-10-30
NZ295303A (en) 1999-07-29
MX9703556A (en) 1997-08-30

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