US 3017325 A
Description (OCR text may contain errors)
3,017,325 THERAPEUTIC COMPOSITION FOR TOPICAL AP- PLICATION AND METHOD OF APPLYING THE SAME David A. Vogel, 3910 N. Janssen, Chicago, Ill. No Drawing. Filed Dec. 29, 1955, Ser. No. 556,083
23 Claims. (Cl. 167-58) This invention relates to therapeutic preparations and more particularly to therapeutic preparations including cytochrome as an active therapeutic agent.
It is an object of the present invention to produce an improved therapeutic composition and more particularly a therapeutic composition for topical application that is effective in the control, treatment and cure of lesions.
Another object of the invention is to provide a therapeutic composition which stimulates and particularly promotes formation of healthy granulation tissue in lesions.
Yet another object of the invention is to provide a therapeutic agent in the form of a viscous material which is substantially non-flowing at room temperature for topical application that has no deleterious effect upon the cells of the body and yet is effective in the control, treatment and cure of lesions.
In connection with the foregoing object, it is an object of the present invention to produce a therapeutic agent of the type set forth for topical application to wounds which is of isotonic nature.
Still another object of the invention is to provide a therapeutic preparation comprising in combination a therapeutic agent of the type set forth together with other materials which promote and enhance the activity of the therapeutic agent.
Yet another object of the present invention is to provide a therapeutic composition comprising cytochrome and an agent which cooperates with the cytochrome in promoting the control, treatment and cure of lesions.
A further object of the invention is to provide a sterile therapeutic composition which is effective in the control, treatment and cure of lesions.
These and other objects and advantages will appear hereinafter.
It has now been found that certain of the foregoing objects are accomplished by incorporating a cytochrome preparation in a viscous material which is substantially non-flowing at room temperatures.
It has also been found that others of the foregoing objects are accomplished by incorporating a promoter in cytochrome compositions. By a promoter is meant a material which cooperates directly with the cytochrome to provide enhanced therapeutic activity, i.e., activating the cytochrome or material which enhances the therapeutic activity of the cytochrome by combatting conditions which prevent the cytochrome from exerting its maximum therapeutic activity.
The term cytochrome is used in this application to include all of the known forms of this class of pigments and includes specifically cytochrome a, cytochrome a cytochrome a cytochrome a cytochrome b, cytochrome b cytochrome c and others. The chemistry of these compounds is very complex and has not been completely investigated. Cytochrome c has been most thoroughly investigated and is found according to the most recent authorities to consist of four pyrrole nuclei and four carbon atoms forming a closed ring, the pyrrole nuclei and carbon atoms alternating around the ring and having various constituent groups attached thereto. It is believed that all of the cytochromes are structurally similar or can actually be derived from or theoretically 3,017,325 Patented Jan. 16, 1962 could be derived or prepared from cytochrome c. The
structure of cytochrome c is believed to be as follows:
CH3 CH3 no a on CH3 NH2 This molecule can be characterized as a dicysteineporphrin having a long protein chain R and R attached to each of the cysteine radicals, respectively. Each of the proteitn chains has been calculated to contain 96:1 amino acids, of which 21 or 22 are lysine residues. There are nine or 10 free oc-amino groups and probably the molecules contain this many polypeptide chains. The other amino acids and the number thereof present in each protein chain are as follows: histidine, 3; arginine, 2; cysteine, 1; tyrosine, 5; tryptophan, 2; glutaric and aspartic acid, 19; leucine fraction 9; alamine+glycine+valine+hydroxyvaline fraction 33. The iron porphyrin has been illustrated and is the natural occurring product. However, suitable reactions can be utilized to change this nuclear metal to some other metal such as magnesium, iron, silver and the like.
Cytochrome in the reduced condition shows a characteristic absorption spectrum with four bands, the posi tions of which are approximately the same regardless of the source of the cytochrome. These bands appear at the following points: 6019 A., 5665 A., 5502 A., and 5210 A. It is believed that none of the individual pigments can produce all four of these bands but instead a mixture of the pigments results in this characteristic absorption spectrum of cytochrome.
The cytochromes are widely dispersed in both the animal and vegetable living organisms. Cytochrome has been found in striated muscles of animals and birds, in bakers yeast, in molluscs, crustacea, 40"species of insects, various tissues of higher animals, bacteria, various bulbs of plants, potatoes, grains of wheat, barley, oats and stamens of young flowers. The better sources'of cytochrome are pigeon breasts, beef heart, pig shoulder, beef tongue, beef shoulder and yeast. In general, the more active the tissue the greater the cytochrome concentration.
A preferred method of preparing cytochrome for use in the present invention is as follows: Ox heart is freed from fat and ligaments and is then very finely minced. 1,100 grams of the ox heart is mixed with 1,100 cc. of 0.15 N trichloroacetic acid and permitted to stand for two hours at room temperature with occasional stirring. The pH during this step is approximately pH 4. After standing for two hours, the liquid is removed by pressure, treated with sodium hydroxide until the reaction mass has a pH 7, and then centrifuged. The resulting clear liquid contains reduced cytochrome and a small amount of oxyhemoglobin. Ammonium sulfate is then added to precipitate the oxyhemoglobin, 50 grams of ammonium sulfate being added to each 100 cc. of solution. The solution is filtered, again treated with ammonium sulfate (5 grams being added for each 100 cc. of solution), and left over-night in an ice chest. The pH at this point is approximately 4.9. The liquid is then filtered and, while cool, mixed with its volume of 20 containing 0.182 grams of cytochrome with an iron content of 0.34 percent. The cytochrome prepared by this process has an equivalent weight of 16,500 and by osmotic pressure determination approximately a molecular weight of also 16,500.
A modification of this process from which a purer product is obtained is as follows. In this process 120 cc. of the product solution from the process set forth in the preceding paragraph is treated with a slight excess of ammonium sulfate. The pH is then adjusted to a pH of 10 with 20 cc. of ammonia having a speclfic gravity postulated that a portion of the protein is removed in the latter procedure.
prising cytochrome as an active therapeutic ingredient, the following examples are given. It is to be understood, however, that these examples are intended to be illustrative only and that they are not to be construed as imi ing the scope of the invention.
Example I ffCa-rbowax 4,000 "grams-.. 30 Carbowax 1,500 do 32 Propylene Glycol do 30 Water n1l 7 gm. 1 The Carbowaxes are non volatile polyethylene glyools soluble in both water and aromatic hydrocarbons.
The cytochrome used in this preparation is the product obtained by using the modified extraction procedure of Keilen and Har'tree set forth above. This procedure is described in Proceedings Royal Society (London) B, 122, 29s 1937).
The Carbowax 4,000 and the Carbowax 1,500 are melted together by applying heat. The cytochrome is soluble in water and is dissolved in the water which forms part of the composition. The aqueous solution of cytochrome is then mixed with the propylene glycol and the resultant mixture added to the melted Carbowaxes. After suitable stirring to obtain uniform distribution of the cytochrome throughout the base, the product is poured into containers and allowed to cool. Upon cooling, the preparation solidifies to form a viscous mass which is non-flowing at room temperatures.
Preferably, a fresh sample of cytochrome is utilized in making the preparation. It is also desirable to avoid subjecting the cytochrome to high temperatures in preparing the medicament base. Accordingly, the melted mixture of Carbowaxes is cooled to as low a temperature as possible while still maintaining fluidity. It has been found that this mDKtUIe of Carbowaxes can be successfully cooled to about 35 C. before adding the mixture containing the cytochrome and still obtain good distribution of the cytochrome throughout the medicament base. Heating the cytochrome to high temperatures tends to impair its wound healing properties.
The finished preparation is applied directly to the wound to be treated. This preparation is particularly useful in inducing and accelerating healing in ulcerous wounds. In fact, many wounds which would not heal at all before application of this preparation can be quickly and effectively healed by the therapeutic preparation of the present application.
Example II 1 gram of cytochrome employed in the preparation of Example I was thoroughly mixed into grams of lanolin which contained about 5 percent of water. The lanolin had previously been heated over a Water bath at 35 C. until it melted and Was maintained at this temperature during the addition of the cytochrome.
Example III 900 grams of anhydrous lanolin and the 225 grams: of U.S.P. White petrolatum were melted together in a double boiler. In a separate container a solution was made by mixing 10 grams of the cytochrome employed in the preparation of Example I with 50. ml. of water. The cytochrome solution was added to the melted material in the double boiler with vigorous stirring. The preparation was then poured into jars and cooled to room temperature.
Example IV The composition was prepared in accordance with the procedure and using the same ingredients as Example I except that 0.1 gram of cytochrome was used. This composition possesses definite healing accelerating properties.
Example V The composition was prepared in accordance with the procedure and using the same ingredients as Example I except that the following parts by weight were utilized:
This composition possesses definite healing accelerating properties.
Example VI Parts by weight D'ermabase 100 Water 2 Dermabase is a specialized emulsion of fatty acid esters and alcohols. In preparing the composition, the cytochrome is first dissolved in the water. The Dermabase is then melted and thereafter the aqueous solution of cytochrome is added thereto. After suitable mixing and stirring the composition while molten was poured into tubes and jars where it was cooled.
It is to be understood that the various cytochromes described herein as suitable for use in accordance with the present invention may be used alone or in mixtures. In fact, the material usually employed is a mixture of two or more cytochrome compounds.
tained by the modified method described above is cytochrome c.
The cytochromes obtained from this modified process are water soluble. Other derivatives of cytochrome are not water soluble. The present invention contemplates the use of water soluble and water insoluble derivatives as well as various salts and degradation products of these cytochromes. a
It is preferable to employ relatively highly purified forms of cytochromes inthe compositions of the present invention. Attention is particularly directed to this fact in view of the normal occurrence of cytochrome in combination with complex cell materials which tends to result in relatively impure extracts. By relatively highly purified forms of cytochrome is meant cytochromes having a purity of percent or greater. It is preferable that the cytochrome have eliminated or separated therefrom as much as possible of the cellular material and other therapeutically inactive substances which may tend to interfere with the therapeutic properties of the cytochrome. It is to be noted that the cytochrome derived by the methods above described has very high purity in .the order of 90 percent or more.
The water soluble derivatives of cytochrome are preferably incorporated in a composition wherein the base is hydrophilic. The Carbowax ointment base and the lanolin base described above are examples of such hydrophilic bases. If it is desired to use water insoluble derivatives, it is preferred to use non-hydrophil-ic bases such as the solid hydrocarbon bases.
The present invention is particularly directed to medicament bases that are viscous or non-flowing at ordinary room temperatures. The purpose of using such a base is to permit topical application for prolonged periods of time to wounds. Lanolin or wool fat, in a purified either hydrous or anhydrous condition is a preferred medicament base. Instead of the lanolin, however, many equivalent fats and unguents may be employed including for example, lard or Aquaphor (mainly eucerin, which comphises a mixture of esters of isoor oxycholesterol alcohols with principally oleic, carnaubic and myristic acids). The unctuous preparations may also comprise petrolatum, monolene and the like, as well as cholesterols, stearoles, palrnitols and other higher and lower fatty acids and alcohols and their esters, such as triacetin and particularly the more or less viscous alkyl alcohols corresponding to both the higher and lower fatty acids and the glycerol esters of the higher fatty acids, such as glycerol stearate, glycerol palmitate and glycerol oleate. It is preferable to employ substances that are bland and non-irritating. The foregoing materials are moreover normally neutral in the presence of water.
Amounts of cytochrome as small as 0.001 gram per 100 grams of medicament base have been found sufiicient to give a preparation having therapeutic properties. On the other hand, relatively high concentration of cytochrome on the order of 50 percent by weight based on the total weight of the composition has been found efiective and have been found to result in no harmful irritation of the tissues. Thus any desired amount of cytochrome may be present provided it is sufficient to give the composition therapeutic properties. A preferred range of cytochrome concentration is from 0.01 percent to 10 percent by weight of the composition. The most economical range of cytochrome concentration is from 0.01 percent to 3 percent by weight of the final composition.
The cytochrome compositions of the present invention are adapted for use generally in compositions for the control, treatment and cure of lesions, which compositions are viscous and nonflowing at room temperatures whereby to make them useful in topical application for effecting such control, treatment and cure. They are particularly adapted for use in the treatment of ulcerous wounds and to this end are applied locally to the affected amples are given.
area. it is preferable to employ the cytochrome compositions so as to cause the wound to heal from the base, particularly where the wound is deep.
Various promoters may be incorporated with the cytochrome compositions described above to produce a therapeutic agent exhibiting increased therapeutic activity. The promoters to be incorporated in cytochrome compositions in accordance with the present invention include the following groups of materials:
(A) Chemical agents such as the sulfonamides, other organic sulphur compounds, benzoates such as hydroxyand amino-benzoates, aldehydes and amino acids having bactericidal properties, namely, sulfanilamide, sulfathia- Zole, sulfaguanidine, carboxysulfathiazole, sulfadiazine, sulfamerazine, sulfapyridine and the following acids and esters: propyl para-hydroxy benzoate, methyl para-hydroxy benzoate, para-aminobenzoic acid, methyl paraamino benzoate, n-propyl para-amino benzoate, n-amyl para-amino 'benzoate, as well as urea and alloxan.
(13) Anti-biotics, namely penicillins and esters thereof, streptothrycin, bacitracin, sterpt-o-mycin, vivicillin, gramicidin, actinomycin, tyrothricin, subtlin, tyrocidine, kojic acid, terramycin and their salts.
(C) Hormones, including cortisone, gonadotripic hormones, androgenic hormones, stilboestrol.
(D) Pepsin, trypsin, lysozyme.
In general, the promoters may be incorporated in any of the wide variety of compositions in which cytochrome is used in the control, treatment and cure of lesions.
The promoters described above when included in the cytochrome compositions as set forth activate and pro mote the therapeutic functions of the cytochrome.
The invention contemplates the inclusion of a plurality of the foregoing promoters in cytochrome compositions as well as a single individual among these promoters. Improved properties result where a plurality of promoters are used because the various promoters described above exhibit different types of effects.
Referring now particularly to the sulfonamides, it is pointed out that the presence of sulfonamides having bactericidal propenties in the cytochrome compositions does not adversely affect the therapeutic properties of the cytochrome. When a sulfonamide is combined with cytochrome it serves to speed the action of destroying the bacteria and thus gives the cytochrome an opportunity to make its therapeutic functions more quickly apparent. Preferred sulfonamides in accordance with the present invention include sulfanilamide and sulfathiazole used either alone, or better yet, in combination. The foregoing remarks which relate particularly to the sulfonamides are intended to be illustrative with respect to the promoters having bactericidal properties and not as an indication thatthe other promoters of this type do not possess similar properties since such properties are possessed by these promoters,
In contrast to the bactericidal promoters the organic sulfur compounds other than the sulfonamides such as cysteine, glutathione and allantoin function to stimulate the growth of and maintain the tone of epithelial tissue. The stimulation of the growth of epithelial tissue is much improved where one or more of these latter materials are included in the cytochrome compositions. An outstanding result of such stimulation is avoidance of the healing of wounds in such a manner as to leave unsightly scars.
The other promoters described above including hormones and the enzymes act in various ways to improve the cytochrome compositions. For example, digestive enzymes such as pepsin aid in the sloughing of dead tissue, hormones serve as nutrients for growing fibroplast and epithelial cells during healing.
For the purpose of indicating preferred methods of preparing the compositions of the present invention which have promoters incorporated therein, the following ex- It is to be understood, however, that these examples are intended to be illustrative only and are not to be construed as limiting the scope of the invention.
Example VII grams of neutral soap powder (castile) and 2 grams of cytochrome were dissolved in 1000 cc. of physiological salt solution. After the solution was thoroughly mixed it was filtered through a Buchner funnel using a double layer of filter paper. 15 grams of sulfathiazole were uniformly dispersed in the solution. This composition also has powerful bactericidal and therapeutic properties.
Example IX 225 grams of U.S.P. white petrolatum and 900 grams of anhydrous lanolin Were melted together in a double boiler. In a separate vessel a paste was formed by mixing 50 milliliters of water with a quantity of 4 grams of cytochrome. After warming the paste to the temperature of the melted material in the double boiler the paste was added thereto with vigorous stirring. 50 grams of a mixture of equal parts of finely divided sulfanilamide and sulfathiazole was mixed with 50 milliliters of water in yet another vessel. This mixture, after warming to the temperature of the melted material in the double boiler, was added to the mixture in the double boiler with vigorous stirring. Stirring was continued until the mass had cooled to 40 C. It was then poured into tubes and jars. The resulting ointment exhibits powerful bactericidal and therapeutic properties.
Example X 454 grams of white beeswax were melted in a double boiler. 1630 grams of liquid petrolatum were then added. In a separate vessel 2.8 grams of sodium-borate and 6 grams of cytochrome were dissolved in 700 milliliters of water. This solution was then added to the melted mass in the double boiler with constant stirring. In yet another vessel 30 grams of' a mixture of equal parts of finely divided sulfanilamide and sulfathiazole were moistened with 50 milliliters of water and the resulting mixture was added to the mixture in the double boiler gradually with vigorous agitation. Stirring was then continued until the temperature reached 50 C. The composition Was then poured into tubes and jars and allowed to cool.
Example XI A quantity of very finely divided cytochrome comthe control, treatment and cure of such lesions.
Example XII 20 grams of S-carboxy sulfathiazole and 2 grams of cytochrome were dissolved in 1000 milliliters of a 5 percent solution of monosodium dihydrogen phosphate in distilled water.
Example XIII A composition containing 1 gram of cytochrome, 25,000 Oxford units of penicillin and 0.1 gram of vitamin B was mixed into 25 milliliters of diethylene glycol.
0.1 gram of polyoxyalkylene derivative of sorbitan monostearate sold under the name of Tween 60 was then added. The resulting mass was added with stirring to a melted mass consisting of 25 grams of anhydrous lanolin and 50 grams of white petrolatum. Stirring was continued until the mass had cooled to 40 C.
Example XIK Parts by weight Dermabase Water 2 Cytochrome 0.1 Hydrocortisone acetate 0.5
The cytochrome is first dissolved in the water. There after the Dermabase is melted and the aqueous solution of cytochrome and the hydrocortisone acetate added thereto. After thorough mixing and stirring the composition while molten was poured into jars and tubes where it was cooled. This preparation is particularly useful in the treatment of itchy lesions and neurodermatitis.
It has been indicated above that the promoters described herein may be added in general to any preparation including cytochrome in therapeutically effective amounts to achieve the improved results described herein. In this connection it is pointed out that if the cytochrome composition to which the promoter is to be added does not include a solvent for the promoter in sufiicient quantity to dissolve all that is added, then the promoter is dispersed in the cytochrome composition, or in the alternative, a solvent for the promoter may be added provided it is compatible with or readily dispensible in the cytochrome composition. In case the promoter exhibits a tendency to settle out or become concentrated in some part of the composition, it is preferable to add to the composition a small amount of an emulsifying or dispersing agent. Such agents serve to retain the promoter in the uniformly dispersed state in which it is preferably distributed at the time of incorporation. The agents also aid in bringing about such uniform distribution and to this end are preferably added prior to adding the promoter.
Not only may the promoters described above be added to the foregoing compositions in any desired manner proportions and the conditions and extent under which these proportions may be varied would unduly lengthen this disclosure and serve no useful purpose. For this riate amounts of such promoters.
Taking the sulfonamides as an example, it is pointed outthat any substantial portion of a sulfonamide having bactericidal properties will exert an appreciable promotmg effect upon the activity of the cytochrome composition, although the effectiveness varies somewhat depending upon the particular sulfonamide used. Amounts of sulfanilamide as small as 0.5 percent to 1.0 percent or less by weight, based on the weight of the composition, are sufficient to have an appreciable bactericidal effect when the composition is applied to wounds. When a stronger bactericidal action is desired, amounts of sulfanilamide up to 5 percent and in severe cases up to 20 percent or higher, by weight, based on the weight of the composition, are employed. The foregoing proportions,
given with respect to sulfanilamide, will indicate the appropriate proportions of various other sulfonamides to apply in any given case and more generally of various other promoters as well. The preferred proportions of the sulfonamides like the preferred proportions of the promoters generally vary with the nature of the compositions in which the promoter is included. Thus, in the more fluid compositions, particularly the aqueous compositions, the smaller proportions are generally employed, although the larger amounts may be used if desired. The less fluid compositions, such as the ointments and salves, generally contain intermediate amounts but in the case of these also the amounts may be varied as desired. The larger amounts are preferably employed in the powdered compositions but in these as well as in the other types of compositions, the quantity may vary. In determining the amount of a sulfonamide to incorporate in a cytochrome composition, it should be borne in mind that the amount is generally somewhat less than would be suitable were the cytochrome not present.
It is not only possible but preferable to employ lesser amounts of the sulfonamides along with the cytochrome because sulfonamides are toxic in nature and tend to destroy tissue in contrast to the cytochrome which is nontoxic and stimulates the cure of lesions. The primary function of the sulfonarnide is to initiate the destruction of the bacteria and give the cytochrome an opportunity to exert its effect. Thus, it is not only preferable to employ the minimum quantity of sulfonamides, but it is also preferable to avoid the use of cytochrome compositions containing sulfonamides as soon as the course of the infection is definitely arrested or the danger of infection is past.
There have been described above the various proportions of cytochrome which are generally used when promoters are not employed. Promoters tend to make smaller amounts of cytochrome more effective and it will be apparent that the inclusion of promoters will permit the use of smaller quantities of cytochrome without appreciable loss of effectiveness of the composition.
The cytochrome compositions including promoters are adapted for use generally in the control, treatment and cure of lesions. They are particularly adapted for use in the treatment of infections or infectious conditions and to this end are applied locally to the affected area, such as an area of inflamed tissue, or lesion. It is preferable to employ the cytochrome compositions so as to cause the infection to heal from the base, particularly where the infection is relatively deep-seated.
The ointments described above are useful in treating various scalp lesions and as a hair ointment or tonic. Alternatively, the cytochrome may be incorporated in a hair shampoo or hair dyes and bleaches for application in that manner. It further is contemplated that various aqueous solutions of cytochrome and alcoholic aqueous solutions of cytochrome can be used as a hair tonic and scalp lotion.
Aqueous solutions and particularly isotonic saline solutions of cytochrome can be used for treatment of the eye. For example, an eye wash or eye lotion would comprise cytochrome dissolved in a suitable aqueous solution.
Various facial preparations can be provided with cytochrome distributed therein in therapeutically effective amounts. Various aqueous and alcoholic aqueous solutions of cytochrome can be used as face lotions for use after shaving or for use as a cleansing medium. Alternatively, the cytochrome could be incorporated in the shaving soap or cream. Other examples of the use of cytochrome on the face would include cold creams and cleansing creams, as well as lotions, lipsticks, rouge, various cream masks and the like.
Various cytochrome preparations are also effective in treating lesions in various external openings, such as nasal passages and ear passages. For example, an aqueous solu- 10 tion of cytochrome could be used as a nose drops.
It further is contemplated that cytochrome be used as the active ingredient in a spray for use under the arm pits and in depilatories. In this connection, it is further contemplated that the cytochrome be incorporated in suppositories for the treatment of areas best handled in this manner.
In treating the feet it is contemplated that various antiseptics and fungicides can be incorporated therein to deal with various infections encountered on the feet.
Various powdered compositions including dusting powders can have cytochrome incorporated therein in therapeutically reactive amounts. Such powders are useful in direct application to wounds and may have incorporated therein other active ingredients such as the various promoters described above.
This application is a continuation-impart of my copending application Serial No. 252,419, filed October 20, 1951, now abandoned, for Composition For Treating Wounds.
It is apparent that nasal spray or as many widely different embodiments of the invention could be made without departing from the spirit and scope thereof. Accordingly, it is not intended that the invention be limited except as indicated in the appended claims.
1. A composition useful in treating ulcerous wounds, which composition comprises cytochrome distributed in a viscous ointment carrier.
2. A composition useful in treating ulcerous wounds, which composition comprises cytochrome c distributed in a viscous ointment carrier. I
3. A composition useful in treating ulcerous wounds, which composition comprises cytochrome prepared sub- 'stantially according to the extraction procedure of Kielen and Hartree distributed in a viscous ointment carrier.
4. A composition useful in treating ulcerous wounds. which composition comprises a cytochrome-active water soluble derivative of cytochrome distributed in a viscous ointment carrier.
5. A composition useful in treating ulcerous wounds, which composition comprises a cytochrome-active water soluble derivative of cytochrome c distributed in a viscous ointment carrier.
6. A composition useful in treating ulcerous wounds, which composition comprises cytochrome distributed in a substantially iso-tonic, viscous ointment carrier.
7. A composition useful in treating ulcerous wounds, which comprises cytochrome distributed in a viscous ointment carrier, the cytochrome comprising from about 0.001 percent to about 50 percent of the composition by weight.
8. A composition useful in treating ulcerous wounds, which comprises cytochrome distributed in a viscous ointment carrier, the cytochrome comprising from about 0.001 percent to about 3 percent of the composition by weight.
9. A composition useful in treating ulcerous wounds, which comprises cytochrome distributed in a viscous ointment carrier, the cytochrome comprising from about 0.01 percent to about 3 percent of the composition by weight.
10. A composition useful in treating ulcerous wounds, which composition comprises cytochrome c distributed in a viscous ointment carrier, the cytochrome 0 comprising from about 0.01 percent to about 3 percent of the composition by weight.
11. A composition useful in treating ulcerous wounds, which composition comprises a cytochrome-active water soluble derivative of cytochrome distributed in a hydrophilic, viscous ointment carrier.
12. A composition useful in treating ulcerous wounds, which comprises cytochrome distributed in a viscous, polyethylene glycol ointment carrier.
13. A composition useful in treating ulcerous wounds,
which composition comprises cytochrome distributed in a viscous, lanolin ointment carrier.
14. A composition useful in treating ulcerous wounds, which composition comprises a cytochrome-active water soluble derivative of cytochrome c distributed in a hydrophilic, isotonic, viscous ointment carrier, the cytochrome comprising from about 0.01 percent to about 3 percent of the composition by weight.
15. A composition useful in treating ulcerous wounds, Which composition comprises cytochrome together with means consisting essentially of a material which cooperates directly with the cytochrome to enhance the therapeutic activity thereof, said cytochrome and material being distributed in a viscous ointment carrier.
16. A composition useful in treating ulcerous wounds, which composition comprises cytochrome together with a bactericide distributed in a carrier.
17. A dusting powder comprising a quantity of powder having cytochrome distributed throughout the mass thereof of a material which cooperates directly with the cytochrome to enhance the therapeutic activity thereof.
22. A composition for use in treating ulcerous wounds, which comprises cytochrome distributed in an emulsion of fatty acid esters and alcohols.
23. A composition for use in treating ulcerous wounds, which comprises cytochrome c distributed in an emulsion of fatty acid esters and alcohols which is viscous and nonfiowing at ordinary room temperature, the cytochrome c comprising from about 0.01 percent to about 3 percent of the composition by weight.
References Cited in the file of this patent UNITED STATES PATENTS 2,320,479 Sperti June 1, 1943 FOREIGN PATENTS 129,555 Australia Oct. 20, 1948 OTHER REFERENCES Proger et al., Journal of Clinical Investigation, vol. 23 (1944), page 949'.
J.A.M.A., vol. 121, No. 15, April 10, 1943, pp. 1237-8.
Czetsch-Lindenwald et al., Salben, Puder, Externa, Springer-Verlag, Berlin, 3rd ed., 1950, pp. -185, 191- 194.
Carruthers, J. Biol. Chem, vol. 171, 1947, pp. 641-51.
Sulzberger, J.A.M.A., Feb. 7, 1953, pp. 468-72.
Modern Drug Encyclopedia, Drug. PubL, N .Y., 4th ed., 1949, p. 231.
Dorland, The American Illus. Medical Dict., W. B. Saunders Co, Phila., 19th ed., 1943, p. 791.
British J. of Surg., April 1945, pp. 518-522.