US20170071998A1 - Hepatoprotective traditional chinese medicine composition, preparation method and use thereof - Google Patents

Hepatoprotective traditional chinese medicine composition, preparation method and use thereof Download PDF

Info

Publication number
US20170071998A1
US20170071998A1 US15/223,131 US201615223131A US2017071998A1 US 20170071998 A1 US20170071998 A1 US 20170071998A1 US 201615223131 A US201615223131 A US 201615223131A US 2017071998 A1 US2017071998 A1 US 2017071998A1
Authority
US
United States
Prior art keywords
extract
parts
clam
chinese medicine
rhizoma curcumae
Prior art date
Legal status (The legal status 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 status listed.)
Abandoned
Application number
US15/223,131
Inventor
Yong Zhou
Hailong Li
Xiaolei Guo
Chung Wah MA
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Infinitus China Co Ltd
Original Assignee
Infinitus China Co Ltd
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 Infinitus China Co Ltd filed Critical Infinitus China Co Ltd
Assigned to INFINITUS (CHINA) COMPANY LTD. reassignment INFINITUS (CHINA) COMPANY LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: GUO, XIAOLEI, LI, HAILONG, MA, Chung Wah, ZHOU, YONG
Publication of US20170071998A1 publication Critical patent/US20170071998A1/en
Priority to US16/412,638 priority Critical patent/US10799549B2/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • 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)
    • A61K36/63—Oleaceae (Olive family), e.g. jasmine, lilac or ash tree
    • A61K36/638—Ligustrum, e.g. Chinese privet
    • 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
    • A61K35/56—Materials from animals other than mammals
    • A61K35/618—Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs
    • 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)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • 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)
    • A61K36/65—Paeoniaceae (Peony family), e.g. Chinese peony
    • 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)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/906—Zingiberaceae (Ginger family)
    • A61K36/9066—Curcuma, e.g. common turmeric, East Indian arrowroot or mango ginger
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

Definitions

  • the present invention relates to the technical field of traditional Chinese medicine, in particular to a hepatoprotective traditional Chinese medicine composition and a preparation method and use thereof.
  • Liver is an organ with a main function of metabolism in the body and plays a role in deoxidation, glycogen storage, synthesis of secreted proteins and the like inside the body. In the meantime, liver also secretes bile in the digestive system. It is mentioned in “Inner Canon of Huangdi-Plain Questions” that, the liver is to a human what a general officer out of which strategies come is to an army. According to traditional Chinese medicine, the liver, one of the five viscera, is in control of the angry mood, corresponds to wood in the five elements (“Five Elements Doctrine”, a major Chinese medical doctrine. The doctrine holds that the universe and nature are composed of five basic elements, i.e., metal, wood, water, fire and earth, so is the human body.
  • liver nourishment and protection has gradually become an important part of health care.
  • Chinese patent CN104740541A discloses a health-care Chinese medicine powder that protects the liver, detoxifies it and restores its function, which is prepared by water decoction and spray drying from 33 kinds of Chinese herbal medicines including Radix Paeoniae Alba, Fructus Schisandrae Chinensis, Fructus Ligustri Lucidi, Herba Ecliptae, Fructus Lycii, Fructus Mori, Radix Angelicae Sinensis, Fructus Mume, Fructus Citri Sarcodactylis, Fructus Hordei Germinatus, Flos Rosae Chinensis, Rhizoma Curcumae Longae and the like.
  • the Chinese medicine powder disclosed in the Patent CN104740541A comprises up to 33 kinds of Chinese herbal medicines, besides the potential problems about medication safety, its high cost, and complicated process limit large-scale production thereof
  • the object of the present invention is to provide a hepatoprotective traditional Chinese medicine composition, which achieves a significant hepatoprotective efficacy with fewer components, and to provide a process for preparing it and its use.
  • the present invention provides the following technical solutions:
  • a hepatoprotective Chinese medicine composition which is prepared by Fructus Ligustri Lucidi, Herba Ecliptae, Radix Paeoniae Alba, Rhizoma Curcumae Longae and clam.
  • the present invention selects three Chinese medicines, Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba, in combination with the extracts of Rhizoma Curcumae Longae and clam to prepare a Chinese medicine composition having a significant hepatoprotective efficacy through unique procedures.
  • the Chinese medicine composition pays attention to not only the efficacy, but also reasonable combination of raw materials in the composition, and thus an optimal formula is obtained.
  • Fructus Ligustri Lucidi is the dry ripe fruit of Ligustrum lucidum Ait , in the Oleaceae family. As recorded in “Chinese Pharmacopoeia”, it is sweet, bitter and cool in nature. It has the efficacies of nourishing the liver and the kidney, brightening eyes and blackening the hair with a dosage of 6-12 g. Clinically it is used to treat yin-deficiency of the liver and the kidney, dizziness, tinnitus, weak waist and knees, premature graying of the beard and the hair, dim eyes, internal heat, diabetes and hectic fever.
  • Herba Ecliptae is the dry aboveground part of Eclipta prostrata L. in the Asteraceae family. As recorded in “Chinese Pharmacopoeia”, it is sweet, sour and cold in nature. It has the efficacies of nourishing the liver and the kidney, cooling the blood and hemostasis with a dosage of 6-12 g. Clinically it is used to treat yin-deficiency of the liver and the kidney, loose teeth, premature graying of the beard and the hair, dizziness, tinnitus, and weak waist and knees.
  • Radix Paeoniae Alba is the dry root of Paeonia lactiflora Pall. in the Ranunculaceae family. As recorded in “Chinese Pharmacopoeia”, it is bitter, sour and slightly cold in nature, and goes to the liver and the spleen channels. It has the efficacies of nourishing the blood, regulating menstruation, astringing yin, suppressing sweating, nourishing the liver, relieving pain and stabilizing yang in the liver. It is used to treat blood deficiency, sallow complexion, irregular menstruation, spontaneous sweating, night sweating, hypochondriac pain, abdominal pain, limb spasm, headache and dizziness with a dosage of 6-15 g.
  • Rhizoma Curcumae Longae is the dry rhizome of Curcuma longa L. in the Zingiberaceae family. It is pungent, bitter and warm in nature, and goes to the spleen and the liver channels. It has the efficacies of invigorating blood and qi circulation and relieving dysmenorrhea with a dosage of 3-10 g. It is used to treat sharp pain in the chest and the lateral thorax, chest stuffiness, heartache, dysmenorrhea, amenorrhea, pelvic mass, shoulder and arm pain caused by rheumatism and swelling and pain due to trauma.
  • Clam is an ancient liver-nourishing tonic. As recorded in “Compendium of Materia Medica ”, “Claim mainly serves to stimulate appetite, inhibit toxicity of mineral medicines, eliminate sudden fever, brighten the eyes, induce diuresis, relieve alcoholism and treat yellowing of the eyes”.
  • the composition of the present invention is prepared by extracting Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba with ethanol and then mixing with Rhizoma Curcumae Longae extract and clam extract.
  • the Rhizoma Curcumae Longae extract is an alcohol extract and the clam extract is a water extract.
  • the parts by weight of the respective components are:
  • the parts by weight of the respective components are:
  • the parts by weight of the respective components are:
  • the parts by weight of the raw materials of the traditional Chinese medicine composition may also be selected as follows:
  • Rhizoma Curcumae Longae extract is obtained by the following process:
  • Rhizoma Curcumae Longae extract is obtained by the following process:
  • the clam extract is obtained by the following process:
  • the clam extract is obtained by the following process:
  • the spraying agent is selected from excipients conducive to the smooth spraying such as maltodextrin, isomalto oligosaccharide powder, ⁇ -cyclodextrin, starch and the like.
  • the Rhizoma Curcumae Longae extract can be purchased from Ningbo Traditional Chinese Medicine Pharmaceutical Co. Ltd.
  • the clam extract can be purchased from Guangzhou Jingfang Pharmaceutical Technology Development Co. Ltd./Guangzhou Mandy Biotechnology Co. Ltd. (produced by Lichuan Farm Co. Ltd.).
  • the Chinese medicine composition of the present invention can inhibit weight loss, significantly reduce ALT (alanine aminotransferase) and ALD (aldosterone), elevate ALB (albumin), increase SOD (superoxide dismutase), reduce MDA (malondialdehyde) and HYP (hydroxyproline), and meantime have a good effect in the evaluation of pathological changes.
  • ALT alanine aminotransferase
  • ALD aldosterone
  • ALB albumin
  • SOD superoxide dismutase
  • MDA malondialdehyde
  • HYP hydroxyproline
  • the health product may be of any dosage form in the current field of health product, and is preferably in the form of oral liquid, capsule, tablet, powder or granule.
  • Each dosage form may be prepared by selecting suitable and acceptable excipients according to practical requirements of the dosage form, which belongs to conventional preparation techniques of dosage forms in the art.
  • the present invention also provides a method of preparing a hepatoprotective health product, comprising providing the traditional Chinese medicine composition as mentioned above.
  • the hepatoprotective health product is in the form of oral liquid, capsule, tablet, powder or granule.
  • the present invention also provides a method for preparing the Chinese medicine composition of the present invention, comprising:
  • the preparation method comprises:
  • the present invention utilizes Fructus Ligustri Lucidi and Herba Ecliptae to nourish the liver blood, Radix Paeoniae Alba and Rhizoma Curcumae Longae to promote Qi flow of the liver, and clam to detoxify the liver. It combines several medicines to nourish the liver, smooth the liver qi and achieves a liver nourishing and protective efficacy, and is applicable to the population suffering from liver injury caused by an unhealthy lifestyle.
  • the Chinese herbal medicine composition of the present invention is characterized in using Fructus Ligustri Lucidi, Herba Ecliptae, Radix Paeoniae Alba, Rhizoma Curcumae Longae and clam as main raw materials. It combines various components reasonably, uses them in conjunction, and achieves a health-care function via multiple means and at multiple levels.
  • FIG. 1 shows an image of liver pathological changes, wherein the arrow indicates hepatocyte steatosis
  • FIG. 2 shows an image of liver pathological changes, wherein arrow 1 indicates pseudolobuli, arrow 2 indicates a small amount of lymphocyte infiltration, and arrow 3 indicates liver fibroplasias;
  • FIG. 3 shows an image of liver pathological changes, wherein the arrow indicates focal lymphocyte infiltration in the portal area;
  • FIG. 4 shows an image of liver pathological changes, wherein arrow 1 indicates bile duct hyperplasia in the portal area, and arrow 2 indicates fibroplasias and inflammatory infiltration in the portal area;
  • FIG. 5 shows an image of liver pathological changes, wherein arrow 1 indicates hemosiderosis and arrow 2 indicates focal lymphocyte infiltration;
  • FIG. 6 shows an image of liver pathological changes, wherein the arrow indicates partial hepatocyte necrosis.
  • the present invention discloses a hepatoprotective traditional Chinese medicine composition and a preparation method and use thereof.
  • Those skilled in the art can carry it out with reference to the contents herein by making suitable improvements to process parameters. It should be particularly noted that all similar replacements and modifications are apparent to those skilled in the art, and are regarded to be within the present invention.
  • the traditional Chinese medicine composition and the preparation method and use thereof in the present invention have been illustrated via preferred examples. Consequently, the method and use described herein can be modified or varied and combined properly by relevant personnel to implement and apply the inventive technique without departing from the contents, spirit and scope of the present invention.
  • Ethanol was added to Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba for extraction.
  • the extract solutions were combined and filtered.
  • the filtrate was concentrated in vacuum into an extractum, which was subjected to spray drying to obtain an alcohol extract of the mixed Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba to be used.
  • Rhizoma Curcumae Longae was ground, and then extracted with ethanol for two times.
  • the extract solutions were combined and concentrated under reduced pressure into an extractum, to which ethanol was added for dissolution.
  • the mixture was filtered.
  • the filtrate was concentrated and dried to obtain a Rhizoma Curcumae Longae extract to be used.
  • the fresh clam was cooked with added water.
  • An extract solution of the clam was obtained after concentration and filtration.
  • the extract solution of the clam was cooled to room temperature, and then mixed well with added maltodextrin and isomalto oligosaccharide powder. The mixture was sterilized and then spray dried to obtain a clam extract to be used.
  • the alcohol extract of the mixed Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba was mixed with the Rhizoma Curcumae Longae extract and clam extract to obtain the traditional Chinese medicine composition.
  • Example 1 The same as Example 1, wherein the Rhizoma Curcumae Longae extract and clam extract were commercially available products.
  • Feeding room animal room in sterile environment is on the second floor of the Experimental Animal Center (north campus), Zhongshan University. Experimental animal usage license No.: SYXK (Yue) 2007-0081.
  • Type Sterilized rat feed.
  • Production unit Guangdong Medical Laboratory Animal Center. Address: 119 Poyang Road, Huang Qi Town, Nanhai District, Foshan City.
  • Feeding method free feed intake.
  • Routine nutrient index of the feed is detected twice a year by Experimental Animal Monitoring Department, Guangdong (with reference to the national standard of China GB14924.3-2010).
  • Feed storage stored in a special feed room and kept ventilative, clean and dry.
  • Type of the drinking water high quality water sterilized at 121° C. (1.0 kg/cm2) for 30 min, which meets “Water Quality Standards for Fine Drinking Water” (CJ94-2005).
  • Water feeding method free intake through animal drinking bottles.
  • Bio-Tek microplate reader Bio-Tek company, U.S.
  • 5417R benchtop refrigerated high-speed centrifuge EPPDORFF company, Germany
  • PL303101 electronic balance Metter-Toled company
  • WFZ800-D3B UV-visible spectrophotometer Ruili Analytical Instrument company, Beijing
  • BIOSPEC handheld tissue homogenizer BIOSPEC company
  • BIO-RAD electrophoresis electroporator BIO-RAD Company, U.S.
  • ImageQuant LAS 4000mini chemiluminescent imager General Electric company, U.S.
  • Carbon tetrachloride (Tianjin Baishi Chemical Industry Co. Ltd., batch number 20130829), hydroxyproline (Hyp), superoxide dismutase (SOD), malondialdehyde (MDA), Coomassie brilliant blue protein kit (all purchased from Nanjing JianCheng Bioengineering Institute with batch numbers 20140711, 20140714, 20140710, 20140725, respectively), NOX2 antibody (Upstate, Millipore, USA), alanine aminotransferase (ALT), albumin (ALB), aldosterone (ALD) ELISA kit (all provided by Shanghai BlueGene Biotech Co. Ltd., batch number 20140813).
  • Wistar rats half male and half female.
  • the animal models were injected subcutaneously with a 40% peanut oil solution of CCl 4 twice a week at 3 ml/kg each time for 5 consecutive weeks. In the 6th week, they were injected once a week instead. In the 5th and 6th weeks, they were additionally intragastrically administrated with the above hyperthermia Chinese medicine at 15 ml/kg once a day for 2 weeks in total.
  • the normal group was injected subcutaneously with the solvent, peanut oil twice a week at 3 ml/kg each time for 5 consecutive weeks. From the 6th week, they were injected once a week instead.
  • control group solvent control
  • model group a 40% peanut oil solution of CCl 4
  • treatment groups A and B high dosage of the sample and low dosage of the sample
  • positive medicine group yiguan decoction group
  • treatment groups C, D and E treatment groups C, D and E.
  • Administration routes and the reason for the selection intragastric administration through the mouth, which was the same as the clinical administration routes.
  • Administration frequency once a day for 2 weeks in total.
  • the body coefficient of human Ka 0.1057
  • the body coefficient of rats Kb 0.086
  • the standard weight of human Wa was calculated as 70 kilograms
  • the weight of rats Wb was calculated as a weight of about 0.25 kilogram at our current stage
  • the dosage for human was D(a)
  • the dosage for rats was D(b).
  • the aconitum carmichaeli, cinnamon and dried ginger (1:1:1) were extracted with water. Specifically, the aconitum carmichaeli was firstly decocted for 1 h, then cinnamon and dried ginger were added to decoct together for 20 min. The decoction was concentrated to 1.2 g of crude medicines per milliliter. The obtained mixture was intragastrically administered once a day at an administration volume of 1.5 ml/100 g.
  • the dosage was 70 g/70 kg/day of the crude medicine, namely 100 mg/100 g/day for human, corresponding to 530 mg/100 g/day for rats.
  • Radix Angelicae Sinensis origin: Gansu, batch number: 140101; Adenophora stricta , origin: Shandong, batch number: YPA4B0001; MeLia toosendan, batch number: YPA2K0001; Ophiopogon japonicus , origin: Hubei, batch number: 140401; Lycium barbarum, batch number: YPA4A0007; Dried rehamnnia root, batch number: YPA3J0001. They were purchased from Caizhilin Pharmacy, Guangzhou Pharmaceutical Group.
  • the compound formula of MeLia toosendan, Radix Angelicae Sinensis, Adenophora stricta, Ophiopogon japonicus, Lycium barbarum and Dried rehamnnia root (1:1.8:1.8:1.8:4:3.6) was extracted with water. Specifically, they were soaked with cold water for 30 min, and then decocted twice for 45 min and 30 min, respectively. The combined decoctions were concentrated until 0.9 g of the crude medicines per milliliter was achieved. It was administered with 0.75 ml/100 g, corresponding to an administration dosage of the crude medicine of 0.675 g/100 g, approximately 1 time the clinical administration dosage.
  • A the dry extract powder obtained by alcohol extraction of the medicinal materials of Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba and drying was formulated into tablets together with the clam extract and Rhizoma Curcumae Longae extract (the traditional Chinese medicine composition of Example 4) after addition of 35% dextrin.
  • the recommended dosage for human is 3.6 g/day, which, after conversion, corresponds to an administration dosage for rats of 27.34 mg/100 g/day as the low dosage group.
  • the dosage in the high dosage group A was 3 times as much as that in the low dosage: 81 mg/100 g/d.
  • Group C the dry extract powder obtained by alcohol extraction of Fructus Ligustri Lucidi and Herba Ecliptae and vacuum drying. The administration dosage for rats was set to 18.11 mg/100 g/day.
  • Group D the dry extract powder obtained by vacuum drying of the alcohol extract of Radix Paeoniae Alba .
  • the administration dosage for rats was set to 4.53 mg/100 g/day.
  • Group E the clam extract and Rhizoma Curcumae Longae extract in Example 4 (8:5).
  • the administration dosage for rats was set to 10.53 mg/100 g/day.
  • the rats were intraperitoneally injected with 10% urethane to anesthetize them. Their abdominal cavities were dissected rapidly. Blood was taken from the abdominal veins with a sterile syringe and put in an ordinary blood tube. The blood was centrifuged at 3000r ⁇ min ⁇ 1 at a low temperature (2-8° C.) for 15 min. The plasma was stored at ⁇ 20° C. till detection. The left lobe of the liver was fixed in 10% formalin. In addition, liver tissue was taken and stored in a refrigerator at ⁇ 80° C. till the detection.
  • ALT Alanine aminotransferase
  • ALB albumin
  • ABD aldosterone
  • a liver tissue was taken, weighed, and then added with an appropriate amount of physiological saline.
  • a 10% homogenate was prepared with a homogenizer in an ice bath.
  • the total protein content in the homogenate of the liver tissue was determined by the Coomassie Brilliant Blue method.
  • Malondialdehyde (MDA) was determined by TBA method and superoxide dismutase (SOD) by xanthine oxidase method according to instructions of the kit.
  • hydroxyproline (HYP) was determined by alkaline hydrolysis of the sample according to instructions of the kit.
  • liver tissue was fixed with 10% formalin, dehydrated, paraffin embeded, sectioned and subjected to hematoxylin-eosin staining (HE) for the pathological morphology examination.
  • HE hematoxylin-eosin staining
  • the low and high dosage treatment groups A, the high dosage treatment group B and the positive control group had an inhibitory effect on the ALT increase with a significant difference.
  • the high dosage treatment group A, the high dosage treatment group B and the positive control group had an inhibitory effect on the SOD decrease with a significant difference.
  • the rest groups had an inhibitory effect on the SOD decrease.
  • the low and high dosage treatment groups A, the high dosage treatment group B, the treatment group C and the positive control group had an inhibitory effect on the MDA increase with a significant difference.
  • the rest groups had an inhibitory effect on the MDA increase.
  • HYP content in the liver tissue in the model group was increased.
  • the low and high dosage treatment groups A, the low and high dosage treatment groups B and the positive control group had an inhibitory effect on the increase of the HYP content with a significant difference.
  • the rest groups had an inhibitory effect on the increase of the HYP content.
  • Main pathologic changes include: bile duct hyperplasia, liver fibrosis, pseudolobuli formation, hepatocyte steatosis, hemosiderosis, lymphocyte infiltration in the portal area, partial hepatocyte necrosis and the like (as shown in FIGS. 1-6 ). The scores of each pathologic change were accumulated. The experimental results were seen in Table 5.
  • the model group had an average score of the pathologic changes differing greatly from that of the blank group, indicating a more serious liver injury of the model group.
  • the lower the score the better the efficacy.
  • the results demonstrate that, all the compositions A, B, C, D and E had a certain alleviating effect on the chemical liver injury.
  • the high dosage group B had a lower score compared to other groups, the score had the largest standard deviation.
  • the traditional Chinese medicine composition in this experiment comprised Fructus Ligustri Lucidi, Herba Ecliptae, Radix Paeoniae Alba , clam extract and Rhizoma Curcumae Longae extract.
  • Each medicine exhibited antioxidative and hepatoprotective effects. They constituted a reasonable composition to effectively alleviate liver injury, protect liver function and alleviate/postpone the occurrence of liver fibrosis.
  • the treatment group A and the treatment group B had a protective effect on liver injury caused by carbon tetrachloride in combination with hyperthermia traditional Chinese medicines and an inhibitory effect on liver fibrosis, and the treatment group A had a better effect.
  • the partial formulations B, C and D had a certain alleviating effect on liver injury caused by carbon tetrachloride in combination with hyperthermia traditional Chinese medicines.

Abstract

Disclosed is a hepatoprotective traditional Chinese medicine composition and a preparation method and use thereof. The composition is prepared by Fructus Ligustri Lucidi, Herba Ecliptae, Radix Paeoniae Alba, Rhizoma Curcumae Longae and clam. The present invention utilizes Fructus Ligustri Lucidi and Herba Ecliptae to nourish the liver blood, Radix Paeoniae Alba and Rhizoma Curcumae Longae to invigorate the liver qi circulation, and clam to detoxify the liver. It combines several medicines to nourish the liver, smooth the liver qi and achieves a liver nourishing and protective efficacy, and is applicable to the population suffering from liver injury caused by an unhealthy lifestyle. The composition is characterized in using Fructus Ligustri Lucidi, Herba Ecliptae, Radix Paeoniae Alba, Rhizoma Curcumae Longae and clam as main raw materials. It combines various components reasonably, uses them in conjunction, and achieves a health-care function via multiple means and at multiple levels.

Description

    CROSS REFERENCE OF RELATED APPLICATION
  • This application claims the benefit of priority to Chinese Patent Application No. 201510577856.X filed on Sep. 11, 2015. The entire content of the above-referenced disclosure is specifically incorporated herein by reference.
    • FIELD
  • The present invention relates to the technical field of traditional Chinese medicine, in particular to a hepatoprotective traditional Chinese medicine composition and a preparation method and use thereof.
    • BACKGROUND
  • Liver is an organ with a main function of metabolism in the body and plays a role in deoxidation, glycogen storage, synthesis of secreted proteins and the like inside the body. In the meantime, liver also secretes bile in the digestive system. It is mentioned in “Inner Canon of Huangdi-Plain Questions” that, the liver is to a human what a general officer out of which strategies come is to an army. According to traditional Chinese medicine, the liver, one of the five viscera, is in control of the angry mood, corresponds to wood in the five elements (“Five Elements Doctrine”, a major Chinese medical doctrine. The doctrine holds that the universe and nature are composed of five basic elements, i.e., metal, wood, water, fire and earth, so is the human body. Only when the five elements in body works in harmony as a whole can health be reached), its position in guts looks very much like spring in the four seasons, and it is closely related to the gallbladder in the six entrails. The liver is responsible for catharsis and is in control of the angry mood. It is also in control of storing blood and is vividly called “a sea of blood” of human body. It is closely linked to human spirits & emotions, digestion & absorption, running of blood & qi (a metaphysical concept in traditional Chinese medicine (TCR). Its amount in body is positively related to the health, according to TCR), fluid metabolism, sex and reproduction.
  • Among modern professionals pursuing career success, there are general phenomena of fast-paced life, high work pressure and frequent staying up late to work overtime and the like, which readily cause blood shortage in the liver, thus bringing people into a sub-health state such as “uncontrollable fatigue”, “no relief no matter how long you have a rest”. Meanwhile, the early-stage diseases or disorders concerning the liver, which are not taken seriously, will cause greater harm later, and may even develop into liver fibrosis, liver cirrhosis, liver cancer, etc. In China, about 58.18% of the population are in a sub-health state, wherein up to 130 million people suffer from liver injury, and nearly 500,000 people die of diseases associated with liver injury. With the growing concerns over health issues, people have realized that once a problem appears in the liver, various subsequent symptoms naturally appear in the body. Therefore, liver nourishment and protection has gradually become an important part of health care.
  • Currently, there have been various types of hepatoprotective products on the market. However, Chinese herbal medicines are highly recognized in the population and favored by the Chinese people due to their unique advantages in liver nourishment and protection. Chinese patent CN104740541A, for example, discloses a health-care Chinese medicine powder that protects the liver, detoxifies it and restores its function, which is prepared by water decoction and spray drying from 33 kinds of Chinese herbal medicines including Radix Paeoniae Alba, Fructus Schisandrae Chinensis, Fructus Ligustri Lucidi, Herba Ecliptae, Fructus Lycii, Fructus Mori, Radix Angelicae Sinensis, Fructus Mume, Fructus Citri Sarcodactylis, Fructus Hordei Germinatus, Flos Rosae Chinensis, Rhizoma Curcumae Longae and the like. Compared to western medicines, Chinese medicines have lower side effects, but the side effects will correspondingly increase with the increase of the Chinese medicine components. Meanwhile, the medication safety cannot be guaranteed as the Chinese medicine components increase. The Chinese medicine powder disclosed in the Patent CN104740541A comprises up to 33 kinds of Chinese herbal medicines, besides the potential problems about medication safety, its high cost, and complicated process limit large-scale production thereof
    • SUMMARY
  • In view of the above mentioned example, the object of the present invention is to provide a hepatoprotective traditional Chinese medicine composition, which achieves a significant hepatoprotective efficacy with fewer components, and to provide a process for preparing it and its use.
  • To achieve the above object, the present invention provides the following technical solutions:
  • A hepatoprotective Chinese medicine composition, which is prepared by Fructus Ligustri Lucidi, Herba Ecliptae, Radix Paeoniae Alba, Rhizoma Curcumae Longae and clam.
  • According to Chinese medicine theories, the present invention selects three Chinese medicines, Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba, in combination with the extracts of Rhizoma Curcumae Longae and clam to prepare a Chinese medicine composition having a significant hepatoprotective efficacy through unique procedures. The Chinese medicine composition pays attention to not only the efficacy, but also reasonable combination of raw materials in the composition, and thus an optimal formula is obtained.
  • Among the selected Chinese medicine components, Fructus Ligustri Lucidi is the dry ripe fruit of Ligustrum lucidum Ait, in the Oleaceae family. As recorded in “Chinese Pharmacopoeia”, it is sweet, bitter and cool in nature. It has the efficacies of nourishing the liver and the kidney, brightening eyes and blackening the hair with a dosage of 6-12 g. Clinically it is used to treat yin-deficiency of the liver and the kidney, dizziness, tinnitus, weak waist and knees, premature graying of the beard and the hair, dim eyes, internal heat, diabetes and hectic fever.
  • Herba Ecliptae is the dry aboveground part of Eclipta prostrata L. in the Asteraceae family. As recorded in “Chinese Pharmacopoeia”, it is sweet, sour and cold in nature. It has the efficacies of nourishing the liver and the kidney, cooling the blood and hemostasis with a dosage of 6-12 g. Clinically it is used to treat yin-deficiency of the liver and the kidney, loose teeth, premature graying of the beard and the hair, dizziness, tinnitus, and weak waist and knees.
  • Radix Paeoniae Alba is the dry root of Paeonia lactiflora Pall. in the Ranunculaceae family. As recorded in “Chinese Pharmacopoeia”, it is bitter, sour and slightly cold in nature, and goes to the liver and the spleen channels. It has the efficacies of nourishing the blood, regulating menstruation, astringing yin, suppressing sweating, nourishing the liver, relieving pain and stabilizing yang in the liver. It is used to treat blood deficiency, sallow complexion, irregular menstruation, spontaneous sweating, night sweating, hypochondriac pain, abdominal pain, limb spasm, headache and dizziness with a dosage of 6-15 g.
  • Rhizoma Curcumae Longae is the dry rhizome of Curcuma longa L. in the Zingiberaceae family. It is pungent, bitter and warm in nature, and goes to the spleen and the liver channels. It has the efficacies of invigorating blood and qi circulation and relieving dysmenorrhea with a dosage of 3-10 g. It is used to treat sharp pain in the chest and the lateral thorax, chest stuffiness, heartache, dysmenorrhea, amenorrhea, pelvic mass, shoulder and arm pain caused by rheumatism and swelling and pain due to trauma.
  • Clam is an ancient liver-nourishing tonic. As recorded in “Compendium of Materia Medica”, “Claim mainly serves to stimulate appetite, inhibit toxicity of mineral medicines, eliminate sudden fever, brighten the eyes, induce diuresis, relieve alcoholism and treat yellowing of the eyes”.
  • Preferably, the composition of the present invention is prepared by extracting Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba with ethanol and then mixing with Rhizoma Curcumae Longae extract and clam extract. The Rhizoma Curcumae Longae extract is an alcohol extract and the clam extract is a water extract.
  • Preferably, the parts by weight of the respective components are:
  • 4-30 parts of Fructus Ligustri Lucidi, 2-15 parts of Herba Ecliptae, 2-15 parts of Radix Paeoniae Alba, 2-20 parts of the Rhizoma Curcumae Longae extract, and 2-10 parts of the clam extract.
  • Further preferably, the parts by weight of the respective components are:
  • 10-25 parts of Fructus Ligustri Lucidi, 5-15 parts of Herba Ecliptae, 3-12 parts of Radix Paeoniae Alba, 6-18 parts of the Rhizoma Curcumae Longae extract, and 4-8 parts of the clam extract.
  • More preferably, the parts by weight of the respective components are:
  • 20 parts of Fructus Ligustri Lucidi, 15 parts of Herba Ecliptae, 8 parts of Radix Paeoniae Alba, 12 parts of the Rhizoma Curcumae Longae extract, and 8 parts of the clam extract.
  • Meantime, in the specific embodiments of the present invention, the parts by weight of the raw materials of the traditional Chinese medicine composition may also be selected as follows:
  • 20 parts of Fructus Ligustri Lucidi, 15 parts of Herba Ecliptae, 10 parts of Radix Paeoniae Alba, 10 parts of the Rhizoma Curcumae Longae extract, and 6 parts of the clam extract.
  • 30 parts of Fructus Ligustri Lucidi, 5 parts of Herba Ecliptae, 5 parts of Radix Paeoniae Alba, 20 parts of the Rhizoma Curcumae Longae extract, and 5 parts of the clam extract.
  • 5 parts of Fructus Ligustri Lucidi, 15 parts of Herba Ecliptae, 15 parts of Radix Paeoniae Alba, 2 parts of the Rhizoma Curcumae Longae extract, and 10 parts of the clam extract.
  • 10 parts of Fructus Ligustri Lucidi, 5 parts of Herba Ecliptae, 15 parts of Radix Paeoniae Alba, 5 parts of the Rhizoma Curcumae Longae extract, and 5 parts of clam extract.
  • 10 parts of Fructus Ligustri Lucidi, 15 parts of Herba Ecliptae, 10 parts of Radix Paeoniae Alba, 5 parts of Rhizoma Curcumae Longae extract, and 10 parts of clam extract.
  • Preferably, the Rhizoma Curcumae Longae extract is obtained by the following process:
  • Grinding the Rhizoma Curcumae Longae, then extracting it with ethanol, concentrating the extract solution underreduced pressure into an extractum, adding ethanol to dissolve the extractum, filtering, concentrating the filtrate, and drying to obtain the Rhizoma Curcumae Longae extract.
  • More preferably, the Rhizoma Curcumae Longae extract is obtained by the following process:
  • Grinding the Rhizoma Curcumae Longae, then extracting it with ethanol twice, combining the two extract solutions, concentrating it under reduced pressure into an extractum, adding ethanol to dissolve it, filtering, concentrating the filtrate, and drying to obtain the Rhizoma Curcumae Longae extract.
  • Preferably, the clam extract is obtained by the following process:
  • Washing the fresh clam, then boiling it with water, concentrating and filtering to obtain an extract solution of the clam, sterilizing, and then drying to obtain the clam extract.
  • More preferably, the clam extract is obtained by the following process:
  • Washing the fresh clam, then boiling it with water, concentrating and filtering to obtain an extract solution of the clam, cooling it to room temperature, then mixing it well with a spraying agent, sterilizing, and then spray drying to obtain the clam extract. Wherein, the spraying agent is selected from excipients conducive to the smooth spraying such as maltodextrin, isomalto oligosaccharide powder, β-cyclodextrin, starch and the like.
  • In addition to the corresponding extracts obtained by the processes for extracting Rhizoma Curcumae Longae and clam defined in the present invention, commercially available corresponding extracts can also be directly used in the present invention. For example, the Rhizoma Curcumae Longae extract can be purchased from Ningbo Traditional Chinese Medicine Pharmaceutical Co. Ltd., and the clam extract can be purchased from Guangzhou Jingfang Pharmaceutical Technology Development Co. Ltd./Guangzhou Mandy Biotechnology Co. Ltd. (produced by Lichuan Farm Co. Ltd.).
  • In the hepatoprotective experiment of rat models of liver injury caused by carbon tetrachloride in combination with a hyperthermia Chinese medicine, compared to the model group, the Chinese medicine composition of the present invention can inhibit weight loss, significantly reduce ALT (alanine aminotransferase) and ALD (aldosterone), elevate ALB (albumin), increase SOD (superoxide dismutase), reduce MDA (malondialdehyde) and HYP (hydroxyproline), and meantime have a good effect in the evaluation of pathological changes. Based on the above technical effects, the present invention provides use of the Chinese medicine composition in the preparation of a hepatoprotective health product.
  • The health product may be of any dosage form in the current field of health product, and is preferably in the form of oral liquid, capsule, tablet, powder or granule. Each dosage form may be prepared by selecting suitable and acceptable excipients according to practical requirements of the dosage form, which belongs to conventional preparation techniques of dosage forms in the art.
  • The present invention also provides a method of preparing a hepatoprotective health product, comprising providing the traditional Chinese medicine composition as mentioned above.
  • In some embodiments, the hepatoprotective health product is in the form of oral liquid, capsule, tablet, powder or granule.
  • Additionally, the present invention also provides a method for preparing the Chinese medicine composition of the present invention, comprising:
      • step 1, extracting Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba with ethanol and concentrating the extract solution to obtain an alcohol extract of the three components;
      • step 2, mixing the alcohol extract of Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba with the Rhizoma Curcumae Longae extract and clam extract to obtain the Chinese medicine composition.
  • More preferably, the preparation method comprises:
      • step 1, extracting Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba with added ethanol, combining the extract solutions, filtrating, concentrating the filtrate in vacuum into an extractum, and spray-drying to obtain the alcohol extract of the mixed Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba;
      • step 2, mixing the alcohol extract of the mixed Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba with the Rhizoma Curcumae Longae extract and clam extract to obtain the Chinese medicine composition.
  • The present invention utilizes Fructus Ligustri Lucidi and Herba Ecliptae to nourish the liver blood, Radix Paeoniae Alba and Rhizoma Curcumae Longae to promote Qi flow of the liver, and clam to detoxify the liver. It combines several medicines to nourish the liver, smooth the liver qi and achieves a liver nourishing and protective efficacy, and is applicable to the population suffering from liver injury caused by an unhealthy lifestyle. The Chinese herbal medicine composition of the present invention is characterized in using Fructus Ligustri Lucidi, Herba Ecliptae, Radix Paeoniae Alba, Rhizoma Curcumae Longae and clam as main raw materials. It combines various components reasonably, uses them in conjunction, and achieves a health-care function via multiple means and at multiple levels.
    • BRIEF DESCRIPTION OF THE DRAWINGS
  • FIG. 1 shows an image of liver pathological changes, wherein the arrow indicates hepatocyte steatosis;
  • FIG. 2 shows an image of liver pathological changes, wherein arrow 1 indicates pseudolobuli, arrow 2 indicates a small amount of lymphocyte infiltration, and arrow 3 indicates liver fibroplasias;
  • FIG. 3 shows an image of liver pathological changes, wherein the arrow indicates focal lymphocyte infiltration in the portal area;
  • FIG. 4 shows an image of liver pathological changes, wherein arrow 1 indicates bile duct hyperplasia in the portal area, and arrow 2 indicates fibroplasias and inflammatory infiltration in the portal area;
  • FIG. 5 shows an image of liver pathological changes, wherein arrow 1 indicates hemosiderosis and arrow 2 indicates focal lymphocyte infiltration;
  • FIG. 6 shows an image of liver pathological changes, wherein the arrow indicates partial hepatocyte necrosis.
    •  DETAILED DESCRIPTION
  • The present invention discloses a hepatoprotective traditional Chinese medicine composition and a preparation method and use thereof. Those skilled in the art can carry it out with reference to the contents herein by making suitable improvements to process parameters. It should be particularly noted that all similar replacements and modifications are apparent to those skilled in the art, and are regarded to be within the present invention. The traditional Chinese medicine composition and the preparation method and use thereof in the present invention have been illustrated via preferred examples. Apparently, the method and use described herein can be modified or varied and combined properly by relevant personnel to implement and apply the inventive technique without departing from the contents, spirit and scope of the present invention.
  • The hepatoprotective traditional Chinese medicine composition and the preparation method and use thereof provided in the present invention are further illustrated as follows to demonstrate the present invention. It is necessary to point out that the following specific examples are only for further demonstrating the present invention, and should not be deemed as restricting the protection scope of the present invention. The non-substantial modification or adjustment made by others according to the present invention still falls into the protection scope of the present invention.
    • Example 1 The Traditional Chinese Medicine Composition of the Present Invention
  • 1. Raw Materials
  • 20 parts of Fructus Ligustri Lucidi, 15 parts of Herba Ecliptae, 10 parts of Radix Paeoniae Alba, 10 parts of Rhizoma Curcumae Longae extract, and 6 parts of clam extract.
  • 2. Preparation Method
  • Ethanol was added to Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba for extraction. The extract solutions were combined and filtered. The filtrate was concentrated in vacuum into an extractum, which was subjected to spray drying to obtain an alcohol extract of the mixed Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba to be used.
  • The Rhizoma Curcumae Longae was ground, and then extracted with ethanol for two times. The extract solutions were combined and concentrated under reduced pressure into an extractum, to which ethanol was added for dissolution. The mixture was filtered. The filtrate was concentrated and dried to obtain a Rhizoma Curcumae Longae extract to be used.
  • After washing, the fresh clam was cooked with added water. An extract solution of the clam was obtained after concentration and filtration. The extract solution of the clam was cooled to room temperature, and then mixed well with added maltodextrin and isomalto oligosaccharide powder. The mixture was sterilized and then spray dried to obtain a clam extract to be used.
  • The alcohol extract of the mixed Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba was mixed with the Rhizoma Curcumae Longae extract and clam extract to obtain the traditional Chinese medicine composition.
    • Example 2 The Traditional Chinese Medicine Composition of the Present Invention
  • 1. Raw Materials
  • 30 parts of Fructus Ligustri Lucidi, 5 parts of Herba Ecliptae, 5 parts of Radix Paeoniae Alba, 20 parts of Rhizoma Curcumae Longae extract, and 5 parts of clam extract.
  • 2. Preparation Method
  • The same as Example 1, wherein the Rhizoma Curcumae Longae extract and clam extract were commercially available products.
    • Example 3 The Traditional Chinese Medicine Composition of the Present Invention
  • 1. Raw Materials
  • 5 parts of Fructus Ligustri Lucidi, 15 parts of Herba Ecliptae, 15 parts of Radix Paeoniae Alba, 2 parts of Rhizoma Curcumae Longae extract, and 10 parts of clam extract.
  • 2. Preparation Method
  • The same as Example 1.
    • Example 4 The Traditional Chinese Medicine Composition of the Present Invention
  • 1. Raw Materials
  • 20 parts of Fructus Ligustri Lucidi, 15 parts of Herba Ecliptae, 8 parts of Radix Paeoniae Alba, 12 parts of Rhizoma Curcumae Longae extract, and 8 parts of clam extract.
  • 2. Preparation Method
  • The same as Example 1.
    • Example 5 The Traditional Chinese Medicine Composition of the Present Invention
  • 1. Raw Materials
  • 10 parts of Fructus Ligustri Lucidi, 5 parts of Herba Ecliptae, 15 parts of Radix Paeoniae Alba, 5 parts of Rhizoma Curcumae Longae extract, and 5 parts of clam extract.
  • 2. Preparation Method
  • The same as Example 1.
    • Example 6 The Traditional Chinese Medicine Composition of the Present Invention
  • 1. Raw Materials
  • 10 parts of Fructus Ligustri Lucidi, 15 parts of Herba Ecliptae, 10 parts of Radix Paeoniae Alba, 5 parts of Rhizoma Curcumae Longae extract, and 10 parts of clam extract.
  • 2. Preparation Method
  • The same as Example 1.
    • Example 7
  • an experiment for demonstrating hepatoprotective efficacy of the traditional Chinese medicine composition of the present invention
  • An experiment for demonstrating the efficacy using the traditional Chinese medicine composition provided in Example 4 above was carried out. Methods and results of the experiment were as follows.
    • 1. EXPERIMENTAL UNIT
  • Pharmacological and toxicological laboratory, School of Pharmacy, Zhongshan University.
    • 2. OBJECT OF THE EXPERIMENT
  • To study the hepatoprotective effect of the traditional Chinese medicine composition of the present invention on rat models of liver injury caused by carbon tetrachloride in combination with hyperthermia traditional Chinese medicines.
    • 3. EXPERIMENTAL ANIMALS
  • Wista rats, half male and half female, weight: females weigh (100-138 g, 118±14 g), males weigh (110-182 g, 150±18 g), provided by Experimental Animal Center, Zhongshan University. Usage license: SYXK (Yue) 2011-0112, production license: SCXK (Yue) 2011-0029.
    • 4. FEEDING MANAGEMENT OF THE ANIMALS
  • Feeding room: animal room in sterile environment is on the second floor of the Experimental Animal Center (north campus), Zhongshan University. Experimental animal usage license No.: SYXK (Yue) 2007-0081.
  • Temperature: 20˜25° C.; humidity: 40%-65%.
    • 5. FEED
  • Type: Sterilized rat feed. Production unit: Guangdong Medical Laboratory Animal Center. Address: 119 Poyang Road, Huang Qi Town, Nanhai District, Foshan City. Production license: SCXK (Yue) 2008-0002.
  • Feeding method: free feed intake.
  • Routine nutrient index of the feed is detected twice a year by Experimental Animal Monitoring Department, Guangdong (with reference to the national standard of China GB14924.3-2010).
  • Feed storage: stored in a special feed room and kept ventilative, clean and dry.
    • 6. DRINKING WATER
  • Type of the drinking water: high quality water sterilized at 121° C. (1.0 kg/cm2) for 30 min, which meets “Water Quality Standards for Fine Drinking Water” (CJ94-2005).
  • Water feeding method: free intake through animal drinking bottles.
    • 7. MAIN EQUIPMENTS AND REAGENTS
  • 7.1 Main Equipments
  • Bio-Tek microplate reader (Bio-Tek company, U.S.), 5417R benchtop refrigerated high-speed centrifuge (EPPDORFF company, Germany), PL303101 electronic balance (Metter-Toled company), WFZ800-D3B UV-visible spectrophotometer (Ruili Analytical Instrument company, Beijing), BIOSPEC handheld tissue homogenizer (BIOSPEC company), BIO-RAD electrophoresis electroporator (BIO-RAD Company, U.S.), ImageQuant LAS 4000mini chemiluminescent imager (General Electric company, U.S.)
  • 7.2 Main Reagents
  • Carbon tetrachloride (Tianjin Baishi Chemical Industry Co. Ltd., batch number 20130829), hydroxyproline (Hyp), superoxide dismutase (SOD), malondialdehyde (MDA), Coomassie brilliant blue protein kit (all purchased from Nanjing JianCheng Bioengineering Institute with batch numbers 20140711, 20140714, 20140710, 20140725, respectively), NOX2 antibody (Upstate, Millipore, USA), alanine aminotransferase (ALT), albumin (ALB), aldosterone (ALD) ELISA kit (all provided by Shanghai BlueGene Biotech Co. Ltd., batch number 20140813).
    • 8. EXPERIMENTAL DESIGN
  • 8.1. Model Preparation
  • Wistar rats, half male and half female. The animal models were injected subcutaneously with a 40% peanut oil solution of CCl4 twice a week at 3 ml/kg each time for 5 consecutive weeks. In the 6th week, they were injected once a week instead. In the 5th and 6th weeks, they were additionally intragastrically administrated with the above hyperthermia Chinese medicine at 15 ml/kg once a day for 2 weeks in total. The normal group was injected subcutaneously with the solvent, peanut oil twice a week at 3 ml/kg each time for 5 consecutive weeks. From the 6th week, they were injected once a week instead.
  • 8.2 Grouping of the Animals
  • Grouping of the experiment: control group (solvent control), model group (a 40% peanut oil solution of CCl4), treatment groups A and B (high dosage of the sample and low dosage of the sample), positive medicine group (yiguan decoction group), and treatment groups C, D and E.
  • 8.3 Administration
  • Administration routes and the reason for the selection: intragastric administration through the mouth, which was the same as the clinical administration routes.
  • Administration frequency: once a day for 2 weeks in total.
  • 8.4 Calculation of Administration Dosages
  • 8.4.1 Conversion of Equivalent Dosages for Rats and for Human

  • D(b)=D(a)*(Kb/Ka)*(Wa/Wb)1/3
  • The body coefficient of human Ka: 0.1057, the body coefficient of rats Kb: 0.086, the standard weight of human Wa was calculated as 70 kilograms, the weight of rats Wb was calculated as a weight of about 0.25 kilogram at our current stage, the dosage for human was D(a), and the dosage for rats was D(b).

  • dosage for rats=dosage for human*(0.086/0.1057)*(70/0.25)1/3=5.3*dosage for human=5.3 times the dosage for human
  • 8.4.2 Hyperthermia Traditional Chinese Medicine
  • 26 g/70 kg·d of the crude medicine for adults, namely 0.371 g/kg/day and 37.1 mg/100 g/day for human, which corresponds to 196 mg/100 g/day after conversion and is taken as 200 mg/100 g/day for rats.
  • Dried ginger, origin: Sichuan, batch number: 1401087; Aconitum carmichaeli, origin: Sichuan, batch number: D1311013; cinnamon: origin: Guangxi, batch number: 1401094. They were purchased from SichuanNew Lotus Chinese Herbal Medicine Co. Ltd.;
  • The aconitum carmichaeli, cinnamon and dried ginger (1:1:1) were extracted with water. Specifically, the aconitum carmichaeli was firstly decocted for 1 h, then cinnamon and dried ginger were added to decoct together for 20 min. The decoction was concentrated to 1.2 g of crude medicines per milliliter. The obtained mixture was intragastrically administered once a day at an administration volume of 1.5 ml/100 g.
  • 8.4.3 Yiguanjian Aconitum
  • As the positive medicine group, the dosage was 70 g/70 kg/day of the crude medicine, namely 100 mg/100 g/day for human, corresponding to 530 mg/100 g/day for rats.
  • Radix Angelicae Sinensis, origin: Gansu, batch number: 140101; Adenophora stricta, origin: Shandong, batch number: YPA4B0001; MeLia toosendan, batch number: YPA2K0001; Ophiopogon japonicus, origin: Hubei, batch number: 140401; Lycium barbarum, batch number: YPA4A0007; Dried rehamnnia root, batch number: YPA3J0001. They were purchased from Caizhilin Pharmacy, Guangzhou Pharmaceutical Group.
  • The compound formula of MeLia toosendan, Radix Angelicae Sinensis, Adenophora stricta, Ophiopogon japonicus, Lycium barbarum and Dried rehamnnia root (1:1.8:1.8:1.8:4:3.6) was extracted with water. Specifically, they were soaked with cold water for 30 min, and then decocted twice for 45 min and 30 min, respectively. The combined decoctions were concentrated until 0.9 g of the crude medicines per milliliter was achieved. It was administered with 0.75 ml/100 g, corresponding to an administration dosage of the crude medicine of 0.675 g/100 g, approximately 1 time the clinical administration dosage.
  • 8.4.4 Treatment Group
  • A: the dry extract powder obtained by alcohol extraction of the medicinal materials of Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba and drying was formulated into tablets together with the clam extract and Rhizoma Curcumae Longae extract (the traditional Chinese medicine composition of Example 4) after addition of 35% dextrin. The recommended dosage for human is 3.6 g/day, which, after conversion, corresponds to an administration dosage for rats of 27.34 mg/100 g/day as the low dosage group. The dosage in the high dosage group A was 3 times as much as that in the low dosage: 81 mg/100 g/d.
  • B: the dry extract powder obtained by alcohol extraction of the medicinal materials of Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba and drying was formulated into tablets after addition of 35% dextrin. The recommended dosage for human is 0.6 g/day, which, after conversion, corresponds to an administration dosage for rats of 4.56 mg/100 g/day as the low dosage group B. The dosage in the high dosage group B was 3 times as much as that in the low dosage group B.
  • Group C: the dry extract powder obtained by alcohol extraction of Fructus Ligustri Lucidi and Herba Ecliptae and vacuum drying. The administration dosage for rats was set to 18.11 mg/100 g/day.
  • Group D: the dry extract powder obtained by vacuum drying of the alcohol extract of Radix Paeoniae Alba. The administration dosage for rats was set to 4.53 mg/100 g/day.
  • Group E: the clam extract and Rhizoma Curcumae Longae extract in Example 4 (8:5). The administration dosage for rats was set to 10.53 mg/100 g/day.
    • 9. EXPERIMENTAL METHOD
  • At the end of the 8th week, the rats were intraperitoneally injected with 10% urethane to anesthetize them. Their abdominal cavities were dissected rapidly. Blood was taken from the abdominal veins with a sterile syringe and put in an ordinary blood tube. The blood was centrifuged at 3000r·min−1 at a low temperature (2-8° C.) for 15 min. The plasma was stored at −20° C. till detection. The left lobe of the liver was fixed in 10% formalin. In addition, liver tissue was taken and stored in a refrigerator at −80° C. till the detection.
  • 9.1 Observation of Symptoms and Signs of the Rats
  • The hair, activity, stool, water intake, weight and the like of the rats in the control group, model group and administration group were observed.
  • 9.2 Detection of Serum Indexes
  • Alanine aminotransferase (ALT), albumin (ALB) and aldosterone (ALD) in the serum samples were determined with an ELISA kit according to the method in the instructions.
  • 9.3 Detection of Liver Indexes
  • A liver tissue was taken, weighed, and then added with an appropriate amount of physiological saline. A 10% homogenate was prepared with a homogenizer in an ice bath. The total protein content in the homogenate of the liver tissue was determined by the Coomassie Brilliant Blue method. Malondialdehyde (MDA) was determined by TBA method and superoxide dismutase (SOD) by xanthine oxidase method according to instructions of the kit. In addition, after weighing of the liver tissue, hydroxyproline (HYP) was determined by alkaline hydrolysis of the sample according to instructions of the kit.
  • 9.4 Pathological Examination of the Liver Tissue
  • The liver tissue was fixed with 10% formalin, dehydrated, paraffin embeded, sectioned and subjected to hematoxylin-eosin staining (HE) for the pathological morphology examination.
  • 9.5 Statistical Processing
  • All data were expressed in the form of mean±standard deviation, analyzed with SPSS10.0 statistical software, and subjected to one-way analysis of variance and a pairwise comparative examination by the LSD method. p<0.05 represents statistical significance.
    • 10. EXPERIMENTAL RESULTS
  • 10.1 Observations of the General State of the Rats
  • No death of the rats occurred in the solvent control group. They were well developed, active and lively with smooth and glossy coat. The animals in the model group were in a poor mental state and less active, with flabby coat and a weight loss (see Table 1).
  • TABLE 1
    Effect on weights of rat models of liver injury caused by CCl4 in
    combination with hyperthermia traditional Chinese medicine
    Group Weight (g) 6th week Weight (g) 8th week
    Solvent control group 315.4 ± 47.5 348.4 ± 47.1
    (n = 9)
    Model group (n = 8) 289.6 ± 47.3 312.5 ± 46.7
    Low dosage group A 279.0 ± 34.9 309.5 ± 47.6
    (n = 8)
    High dosage group A 295.0 ± 57.9 325.1 ± 63.9
    (n = 8)
    Low dosage group B 289.3 ± 41.5 321.3 ± 47.1
    (n = 8)
    High dosage group B 296.5 ± 59.3 311.7 ± 61.6
    (n = 8)
    Positive control group 297.6 ± 48.9 320.5 ± 52.8
    (n = 8)
    Group C (n = 7) 283.1 ± 38.3 303.0 ± 48.3
    Group D (n = 7) 286.4 ± 36.6 300.7 ± 49.6
    Group E (n = 7) 295.8 ± 42.8 311.0 ± 42.7
    # P < 0.05 compared to the solvent control group,
    ## P < 0.01 compared to the solvent control group;
    * P < 0.05 compared to the model group,
    ** P < 0.01 compared to the model group.
  • All of the rats in the model group suffered from a weight loss. After treatment, the high dosage treatment group A and the positive control group had an inhibitory effect on the weight loss, but without significant difference.
  • 10.2 Effect on Serum ALT, ALB and ALD of the Rats
  • TABLE 2
    Effect on serum ALT, ALB and ALD of the rat models of liver injury
    caused by CCl4 in combination with hyperthermia traditional
    Chinese medicine (X ± s)
    Group ALT/U · L1 ALB/g · L1 ALD/ng · L1
    Solvent control group  40.7 ± 13.3 43.9 ± 12.9  76.9 ± 19.0
    (n = 9)
    Model group (n = 8) 131.4 ± 30.4## 28.7 ± 9.3## 121.2 ± 30.8##
    Positive control group 100.4 ± 24.3* 35.4 ± 7.7  91.1 ± 23.8*
    (n = 8)
    Low dosage group A 103.9 ± 22.9* 35.0 ± 12.8  98.5 ± 34.1
    (n = 8)
    High dosage group A  85.3 ± 28.1** 38.7 ± 7.5*  91.5 ± 29.1*
    (n = 8)
    Low dosage group B 107.7 ± 21.2 31.6 ± 9.2 103.1 ± 33.2
    (n = 8)
    High dosage group B  99.5 ± 27.1* 31.2 ± 9.6  99.2 ± 28.8
    (n = 8)
    Treatment group C 111.2 ± 25.9 32.9 ± 8.9 103.7 ± 26.7
    (n = 7)
    Treatment group D 120.3 ± 29.1 26.8 ± 11.6 116.4 ± 24.8
    (n = 7)
    Treatment group E 107.1 ± 28.8 31.2 ± 9.1 104.6 ± 32.5
    (n = 7)
    #P < 0.05 compared to the solvent control group,
    ##P < 0.01 compared to the solvent control group;
    *P < 0.05 compared to the model group,
    **P < 0.01 compared to the model group.
  • Compared to the normal group, serum ALT in the model group was significantly increased. After treatment, the low and high dosage treatment groups A, the high dosage treatment group B and the positive control group had an inhibitory effect on the ALT increase with a significant difference.
  • Compared to the normal group, serum ALB in the model group significantly decreased. After treatment, the high dosage treatment group A had an inhibitory effect on the ALB decrease with a significant difference. The rest groups had an inhibitory effect on the ALB decrease.
  • Compared to the normal group, serum ALD in the model group significantly increased. After treatment, the high dosage treatment group A and the positive control group had an inhibitory effect on the ALD increase with a significant difference. The rest groups had an inhibitory effect on the ALD increase.
  • 10.3 Effect on SOD and MDA in the Liver Homogenate and HYP in the Liver Tissue of Rats
  • TABLE 3
    Effect on SOD and MDA in the liver tissue homogenate and HYP in the
    liver tissue of the rat models of liver injury caused by CCl4 in
    combination with hyperthermia traditional Chinese medicine (X ± s)
    Group SOD/U · mg1 MDA/μmol · g1 HYP/ug · g1
    Solvent control 111.2 ± 27.1  0.62 ± 0.11 119.2 ± 31.1
    group (n = 9)
    Model group  64.2 ± 21.3## 1.37 ± 0.38## 295.1 ± 50.6##
    (n = 8)
    Positive control  93.4 ± 26.1* 0.85 ± 0.25** 222.1 ± 54.1**
    group (n = 8)
    Low dosage 87.1 ± 23.9 1.05 ± 0.34* 234.8 ± 36.9**
    group A (n = 8)
    High dosage  94.6 ± 29.9* 0.91 ± 0.19* 209.4 ± 51.7**
    group A (n = 8)
    Low dosage 84.5 ± 25.2 1.11 ± 0.29 228.9 ± 50.7**
    group B (n = 8)
    High dosage  92.0 ± 22.4* 0.99 ± 0.27* 211.6 ± 57.1**
    group B (n = 8)
    Treatment 85.1 ± 24.4 1.07 ± 0.27* 248.5 ± 38.1
    group C (n = 7)
    Treatment 77.2 ± 21.6 1.10 ± 0.29 257.6 ± 46.8
    group D (n = 7)
    Treatment 76.6 ± 15.8 1.08 ± 0.30 248.8 ± 42.2
    group E (n = 7)
    #P < 0.05 compared to the solvent control group,
    ##P < 0.01 compared to the solvent control group;
    *P < 0.05 compared to the model group,
    **P < 0.01 compared to the model group.
  • Compared to the normal group, SOD in the liver tissue in the model group decreased. After treatment, the high dosage treatment group A, the high dosage treatment group B and the positive control group had an inhibitory effect on the SOD decrease with a significant difference. The rest groups had an inhibitory effect on the SOD decrease.
  • Compared to the normal group, MDA in the liver tissue in the model group was increased. After treatment, the low and high dosage treatment groups A, the high dosage treatment group B, the treatment group C and the positive control group had an inhibitory effect on the MDA increase with a significant difference. The rest groups had an inhibitory effect on the MDA increase.
  • Compared to the normal group, HYP content in the liver tissue in the model group was increased. After treatment, the low and high dosage treatment groups A, the low and high dosage treatment groups B and the positive control group had an inhibitory effect on the increase of the HYP content with a significant difference. The rest groups had an inhibitory effect on the increase of the HYP content.
  • 10.4 Pathological Effect on the Liver Tissue
  • TABLE 4
    Grading standard of histopathologic changes
    Pathological injury range Degree classification Score
    No histologic change Normal 0
    observed
     0%-20% Mild 1
    21%-40% Moderate 2
    41%-100% Severe 3
  • Main pathologic changes include: bile duct hyperplasia, liver fibrosis, pseudolobuli formation, hepatocyte steatosis, hemosiderosis, lymphocyte infiltration in the portal area, partial hepatocyte necrosis and the like (as shown in FIGS. 1-6). The scores of each pathologic change were accumulated. The experimental results were seen in Table 5.
  • TABLE 5
    Scores of pathologic changes
    Score
    Group Mean SD
    Solvent control group (n = 9) 1.71 ± 0.76
    Model group (n = 8) 5.14 ± 1.96
    Low dosage group A (n = 8) 3.71 ± 1.11
    High dosage group A (n = 8) 4.00 ± 0.82
    Low dosage Group B (n = 8) 4.43 ± 1.51
    High dosage group B (n = 8) 3.29 ± 2.06
    Positive control group 3.29 ± 1.38
    Treatment group C (n = 7) 4.00 ± 1.00
    Treatment group D (n = 7) 4.33 ± 1.53
    Treatment group E (n = 7) 4.00 ± 0.00
  • It can be seen from the results that, the model group had an average score of the pathologic changes differing greatly from that of the blank group, indicating a more serious liver injury of the model group. The lower the score, the better the efficacy. The results demonstrate that, all the compositions A, B, C, D and E had a certain alleviating effect on the chemical liver injury. Although the high dosage group B had a lower score compared to other groups, the score had the largest standard deviation.
    • 11. CONCLUSION
  • The traditional Chinese medicine composition in this experiment comprised Fructus Ligustri Lucidi, Herba Ecliptae, Radix Paeoniae Alba, clam extract and Rhizoma Curcumae Longae extract. Each medicine exhibited antioxidative and hepatoprotective effects. They constituted a reasonable composition to effectively alleviate liver injury, protect liver function and alleviate/postpone the occurrence of liver fibrosis. To sum up, the treatment group A and the treatment group B had a protective effect on liver injury caused by carbon tetrachloride in combination with hyperthermia traditional Chinese medicines and an inhibitory effect on liver fibrosis, and the treatment group A had a better effect. The partial formulations B, C and D had a certain alleviating effect on liver injury caused by carbon tetrachloride in combination with hyperthermia traditional Chinese medicines.
  • Only preferred embodiments of the present invention are described above. It should be noted that for those of ordinary skill in the art, several improvements and modifications may also be made without departing from the principle of the present invention, which is also to be regarded as within the protection scope of the present invention.

Claims (10)

1. A hepatoprotective traditional Chinese medicine composition, which is prepared by Fructus Ligustri Lucidi, Herba Ecliptae, Radix Paeoniae Alba, Rhizoma Curcumae Longae and clam.
2. The traditional Chinese medicine composition according to claim 1, which is prepared by extracting Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba with ethanol and then mixing with a Rhizoma Curcumae Longae extract and a clam extract, wherein the Rhizoma Curcumae Longae extract is an alcohol extract of Rhizoma Curcumae Longae and the clam extract is a water extract of clam.
3. The traditional Chinese medicine composition according to claim 2, wherein the parts by weight of the respective components are:
4-30 parts of Fructus Ligustri Lucidi, 2-15 parts of Herba Ecliptae, 2-15 parts of Radix Paeoniae Alba, 2-20 parts of the Rhizoma Curcumae Longae extract, and 2-10 parts of the clam extract.
4. The traditional Chinese medicine composition according to claim 3, wherein the parts by weight of the respective components are:
10-25 parts of Fructus Ligustri Lucidi, 5-15 parts of Herba Ecliptae, 3-12 parts of Radix Paeoniae Alba, 6-18 parts of the Rhizoma Curcumae Longae extract, and 4-8 parts of the clam extract.
5. The traditional Chinese medicine composition according to claim 4, wherein the parts by weight of the respective components are:
20 parts of Fructus Ligustri Lucidi, 15 parts of Herba Ecliptae, 8 parts of Radix Paeoniae Alba, 12 parts of the Rhizoma Curcumae Longae extract, and 8 parts of the clam extract.
6. The traditional Chinese medicine composition according to claim 2, wherein the Rhizoma Curcumae Longae extract is prepared by the following process:
Grinding the Rhizoma Curcumae Longae, then extracting it with ethanol, concentrating the extract solution under reduced pressure into an extractum, adding ethanol to dissolve the extractum, filtering, concentrating the filtrate, and drying to obtain the Rhizoma Curcumae Longae extract.
7. The traditional Chinese medicine composition according to claim 2, wherein the clam extract is prepared by the following process:
Washing the fresh clam, then boiling it with water, concentrating and filtering to obtain an extract solution of the clam, sterilizing, and then drying to obtain the clam extract.
8. A method of preparing a hepatoprotective health product, comprising providing the traditional Chinese medicine composition according to claim 1.
9. The method according to claim 8, wherein the hepatoprotective health product is in the form of oral liquid, capsule, tablet, powder or granule.
10. A method for preparing the traditional Chinese medicine composition according to claim 1, comprising:
step 1, extracting Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba with ethanol and concentrating the extract solution to obtain an alcohol extract of the three components;
step 2, mixing the alcohol extract of Fructus Ligustri Lucidi, Herba Ecliptae and Radix Paeoniae Alba with the Rhizoma Curcumae Longae extract and the clam extract to obtain the traditional Chinese medicine composition.
US15/223,131 2015-09-11 2016-07-29 Hepatoprotective traditional chinese medicine composition, preparation method and use thereof Abandoned US20170071998A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US16/412,638 US10799549B2 (en) 2015-09-11 2019-05-15 Hepatoprotective traditional chinese medicine composition containing fructus ligustri lucidi, herba exliptae, radix paeoniae alba, rhizoma curcumae longae and clam extract, preparation method and use thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201510577856.X 2015-09-11
CN201510577856.XA CN105126026A (en) 2015-09-11 2015-09-11 Traditional Chinese medicinal composition with hepatoprotective effect, and preparation method and application of traditional Chinese medicinal composition

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US16/412,638 Division US10799549B2 (en) 2015-09-11 2019-05-15 Hepatoprotective traditional chinese medicine composition containing fructus ligustri lucidi, herba exliptae, radix paeoniae alba, rhizoma curcumae longae and clam extract, preparation method and use thereof

Publications (1)

Publication Number Publication Date
US20170071998A1 true US20170071998A1 (en) 2017-03-16

Family

ID=54711837

Family Applications (2)

Application Number Title Priority Date Filing Date
US15/223,131 Abandoned US20170071998A1 (en) 2015-09-11 2016-07-29 Hepatoprotective traditional chinese medicine composition, preparation method and use thereof
US16/412,638 Active US10799549B2 (en) 2015-09-11 2019-05-15 Hepatoprotective traditional chinese medicine composition containing fructus ligustri lucidi, herba exliptae, radix paeoniae alba, rhizoma curcumae longae and clam extract, preparation method and use thereof

Family Applications After (1)

Application Number Title Priority Date Filing Date
US16/412,638 Active US10799549B2 (en) 2015-09-11 2019-05-15 Hepatoprotective traditional chinese medicine composition containing fructus ligustri lucidi, herba exliptae, radix paeoniae alba, rhizoma curcumae longae and clam extract, preparation method and use thereof

Country Status (3)

Country Link
US (2) US20170071998A1 (en)
EP (1) EP3141256B1 (en)
CN (1) CN105126026A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110959786A (en) * 2019-11-07 2020-04-07 广东海洋大学 Flavored beverage with liver protecting and sobering functions

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107788438A (en) * 2017-09-20 2018-03-13 南京慧博生物科技有限公司 A kind of Antialcoholic liver-protecting turmeric multifunctional food and preparation method thereof

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004194616A (en) * 2002-12-20 2004-07-15 Creatis Dam:Kk Health food product containing young barley leaf powder extract and fresh water clam extract
JP2004269430A (en) * 2003-03-10 2004-09-30 Ichimaru Pharcos Co Ltd Composition for hair treatment and hair cosmetic for damaged hair
CN1943724B (en) * 2005-10-08 2011-09-21 周小明 A Chinese traditional medicinal composition for treatment of migraine and its preparation method
CN102512600B (en) * 2011-12-13 2013-09-18 常熟雷允上制药有限公司 Health product for protecting liver and preparation method thereof
CN103749825B (en) * 2013-12-31 2015-11-18 张绪伟 Fruit of glossy privet cosmetic tea of a kind of black invention order, nti-freckle and preparation method thereof
US20150190450A1 (en) * 2014-01-05 2015-07-09 Alice Chang Ingredient for consumption and application
CN104474459A (en) * 2014-11-25 2015-04-01 梧州市人民医院 Traditional Chinese medicine preparation with liver protection effect and preparation method thereof
CN104398986A (en) * 2014-11-27 2015-03-11 青岛申达高新技术开发有限公司 Tranquilization and brain tonifying epimedium herb oral liquid and preparation method thereof
CN104740541B (en) 2015-04-21 2019-05-14 杨波 A kind of protecting liver and detoxication restores the Chinese medicine powder for health care of liver function
CN105582471A (en) 2016-01-22 2016-05-18 王平平 Traditional Chinese medicine for treating liver-qi stagnation type premenstrual tension syndrome

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110959786A (en) * 2019-11-07 2020-04-07 广东海洋大学 Flavored beverage with liver protecting and sobering functions

Also Published As

Publication number Publication date
CN105126026A (en) 2015-12-09
EP3141256B1 (en) 2020-05-06
US10799549B2 (en) 2020-10-13
EP3141256A1 (en) 2017-03-15
US20190269746A1 (en) 2019-09-05

Similar Documents

Publication Publication Date Title
CN103990081B (en) Traditional Chinese medicine composition and application thereof in preparation of medicine for treating infant jaundice
CN104491759B (en) A kind of Chinese medicine composition for treating antimigraine and preparation method thereof
CN103520572A (en) Traditional Chinese composition used for treating atopic dermatitis as well as preparation method of composition
CN104958607A (en) Traditional Chinese medicine composition for treating burning mouth syndromes of climacteric women and preparation method thereof
US10799549B2 (en) Hepatoprotective traditional chinese medicine composition containing fructus ligustri lucidi, herba exliptae, radix paeoniae alba, rhizoma curcumae longae and clam extract, preparation method and use thereof
CN104740541A (en) Health-care traditional Chinese medicine powder for protecting liver, detoxifying and recovering liver functions
CN103463297A (en) Traditional Chinese medicine composition for treating eczema of scrotum as well as preparation method and use thereof
CN103432559B (en) Traditional Chinese medicine composition for treating spanomenorrhea and dysmenorrhea caused by blood stasis and preparation method and application thereof
CN109820943B (en) Zhuang medicine preparation for treating gout and preparation method thereof
CN101485786B (en) Whitening spot-eliminating composition and method for preparing the same
CN107890528B (en) Traditional Chinese medicine composition for treating hyperuricemia and preparation method thereof
CN102846871B (en) Chinese herbal preparation for treating liver-fire ascending type supraorbital neuralgia and preparation method thereof
CN106177268A (en) A kind of Chinese medicine composition treating sexual impotence and preparation method thereof
CN105343660A (en) Traditional Chinese medicinal preparation for treating peripheral vertigo and preparation method of traditional Chinese medicinal preparation
CN105902867B (en) A herbal composition for reducing hypertension, hyperglycemia and hyperlipidemia, and its preparation method
CN105169265A (en) Traditional Chinese medicine composition for treating hemiplegia and preparation method of traditional Chinese medicine composition
CN103191297B (en) Traditional Chinese medicine composition for treating male infertility and preparation method thereof
CN103735725B (en) For treating rheumatismal medicine of pig and preparation method thereof
CN102861227B (en) Traditional Chinese medicine composition for treating suppurative meningitis sequelae and preparation method of traditional Chinese medicine composition
CN108175818B (en) Traditional Chinese medicine composition for treating glaucoma and preparation method and application thereof
CN105381410A (en) Traditional Chinese medicinal preparation for treating hepatitis
CN106177404A (en) A kind of Chinese medicine composition treating the hepatic and renal YIN deficiency and preparation method thereof
CN104784558A (en) Traditional Chinese medicine composition for treating recurrent oral ulceration and preparation method thereof
CN104800646A (en) Traditional Chinese medicine combination for treating burning month syndrome and preparation method thereof
CN105327219A (en) Traditional Chinese medicine composition for treating female pelvic inflammation and preparation method of traditional Chinese medicine composition

Legal Events

Date Code Title Description
AS Assignment

Owner name: INFINITUS (CHINA) COMPANY LTD., CHINA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:ZHOU, YONG;LI, HAILONG;GUO, XIAOLEI;AND OTHERS;REEL/FRAME:039288/0305

Effective date: 20160722

STPP Information on status: patent application and granting procedure in general

Free format text: FINAL REJECTION MAILED

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION