WO2021209070A1 - Ginsenoside composition having alcoholic fatty liver-preventing and -treating function - Google Patents

Ginsenoside composition having alcoholic fatty liver-preventing and -treating function Download PDF

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WO2021209070A1
WO2021209070A1 PCT/CN2021/088142 CN2021088142W WO2021209070A1 WO 2021209070 A1 WO2021209070 A1 WO 2021209070A1 CN 2021088142 W CN2021088142 W CN 2021088142W WO 2021209070 A1 WO2021209070 A1 WO 2021209070A1
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ginsenoside
pharmaceutical composition
liver
weight
alcoholic fatty
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PCT/CN2021/088142
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French (fr)
Chinese (zh)
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范代娣
贺婧
段志广
严建亚
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陕西巨子生物技术有限公司
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Priority to AU2021257523A priority Critical patent/AU2021257523B2/en
Publication of WO2021209070A1 publication Critical patent/WO2021209070A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • 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

Definitions

  • the invention relates to the field of biomedicine, in particular to a ginsenoside Rk3/Rh4 composition capable of preventing and treating alcoholic fatty liver.
  • Fatty liver disease is a common disease. It can be divided into alcoholic fatty liver (AFL) and non-alcoholic fatty liver (NAFLD) according to the history of heavy drinking.
  • AFL is caused by long-term alcohol consumption.
  • the main causes of injury are toxic metabolites, oxidative stress and metabolic disorders caused by alcohol metabolism in liver cells.
  • the pathogenesis of NAFLD is mainly related to IR, abnormal liver fat metabolism, mitochondrial dysfunction and oxidative stress, genetic variation and metabolic changes, and susceptibility to cell damage.
  • various liver diseases caused by viral hepatitis occupy a major position, with the rise of alcohol in the human diet, the incidence of alcoholic fatty liver is getting higher and higher, and the chance of developing liver cirrhosis and liver cancer far exceeds NAFLD. It has become the second leading cause of liver damage after viral hepatitis.
  • Polyene phosphatidylcholine is a commonly used drug for adjuvant treatment of alcoholic fatty liver. It can effectively reduce oxygen free radicals, protect liver cell membranes, and delay liver fibrosis, thereby promoting the recovery of patients. Almost cause intestinal disorders and cause diarrhea.
  • Ginsenoside is the active ingredient in ginseng and a sterol compound that can affect multiple metabolic pathways in the body. Studies have reported that ginsenosides Rk3 and Rh4 have obvious anti-tumor, anti-oxidant, anti-inflammatory, and anti-apoptosis effects.
  • ginsenosides CK, Rh1 and their combinations can be used to improve non-alcoholic fatty liver
  • alcoholic fatty liver has not been studied
  • ginsenoside Rg1 has a certain effect in the treatment of metabolic disorders caused by drinking, but whether it has a protective effect on the liver has not been studied
  • ginsenoside Rk3/Rh4 Combination drugs have never been used to improve fatty liver, liver disease and the reduction of liver antioxidants caused by alcohol.
  • the purpose of the present invention is to provide a ginsenoside Rk3/Rh4 composition with the function of preventing and treating alcoholic fatty liver.
  • the composition has good therapeutic and improving effects on alcoholic fatty liver.
  • the Rk3/Rh4 composition can significantly improve the survival rate of drinking mice, reduce serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and reduce serum and liver cholesterol (TC) , Triglyceride (TG), low-density lipoprotein (LDL) content, reduce lipid accumulation (oil droplets) in the liver caused by drinking, liver tissue inflammatory infiltration and fatty vacuoles.
  • ALT serum alanine aminotransferase
  • AST aspartate aminotransferase
  • TC serum and liver cholesterol
  • LDL low-density lipoprotein
  • the present invention includes:
  • a pharmaceutical composition that can be used for the prevention and/or treatment of alcoholic fatty liver, which comprises preventive and/or therapeutically effective amounts of ginsenoside Rk3 and ginsenoside Rh4 as active ingredients, and a pharmaceutically acceptable carrier,
  • the weight ratio of ginsenoside Rk3 and ginsenoside Rh4 is 1:0.5-2, preferably 1:0.9-1.1.
  • the weight ratio of ginsenoside Rk3 and ginsenoside Rh4 is 1:1.
  • the pharmaceutical composition according to item 1 based on 100 parts by weight of total ginsenosides contained in the pharmaceutical composition, the total of ginsenoside Rk3 and ginsenoside Rh4 contained in the pharmaceutical composition is 10 parts by weight above.
  • the pharmaceutical composition according to item 1 based on 100 parts by weight of total ginsenosides contained in the pharmaceutical composition, the total of ginsenoside Rk3 and ginsenoside Rh4 contained in the pharmaceutical composition is 100 parts by weight .
  • ginsenoside Rk3 and ginsenoside Rh4 in the preparation of a pharmaceutical composition for the prevention and/or treatment of alcoholic fatty liver, wherein the weight ratio of the ginsenoside Rk3 and ginsenoside Rh4 in the pharmaceutical composition It is 1:0.5-2, preferably 1:0.9-1.1.
  • the ginsenoside Rk3/Rh4 composition for the prevention and treatment of alcoholic fatty liver of the present invention can be either a medicine or any other preparation form; it can be used for the purpose of treating related diseases, or it can be used to improve or prevent drinking caused by alcohol. Fatty liver.
  • the preparation can be ginsenoside Rk3/Rh4, or a compound preparation composed of ginsenoside Rk3/Rh4 and other components, wherein ginsenoside Rk3/Rh4 is the main component.
  • the ginsenoside Rk3/Rh4 of the present invention can be obtained by extraction, transformation and separation of ginseng or American ginseng, or it can be obtained by chemical synthesis. Specifically, the ginsenoside Rk3/Rh4 can be obtained from the roots, stems, and/or leaves of ginseng or American ginseng by extracting, transforming, and isolating them using prior art processes.
  • Rk3 or Rh4 can increase the survival rate of drinking mice, reduce serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and reduce serum and liver
  • ALT serum alanine aminotransferase
  • AST aspartate aminotransferase
  • TC cholesterol
  • TG triglycerides
  • LDL low-density lipoprotein
  • the present invention also includes:
  • ginsenoside Rk3 in the preparation of a pharmaceutical composition for preventing and/or treating alcoholic fatty liver.
  • the ginsenoside Rk3 contained in the pharmaceutical composition is 5 parts by weight or more, preferably 10 parts by weight Parts by weight or more, more preferably 20 parts by weight or more, more preferably 50 parts by weight or more, more preferably 70 parts by weight or more, more preferably 80 parts by weight or more, more preferably 90 parts by weight or more, more preferably 95 parts by weight or more, more preferably 99 parts by weight Above, more preferably 100 parts by weight.
  • ginsenoside Rh4 in the preparation of a pharmaceutical composition for the prevention and/or treatment of alcoholic fatty liver.
  • the ginsenoside Rh4 contained in the pharmaceutical composition is 5 parts by weight or more, preferably 10 parts by weight Parts or more, more preferably 20 parts by weight or more, more preferably 50 parts by weight or more, more preferably 70 parts by weight or more, more preferably 80 parts by weight or more, more preferably 90 parts by weight or more, more preferably 95 parts by weight or more, more preferably 99 parts by weight Above, more preferably 100 parts by weight.
  • the above-mentioned pharmaceutical composition may be, for example, an oral agent or an injection.
  • the oral agent may be, for example, a hard capsule, a soft capsule, a sustained-release capsule, a sugar-coated tablet, a powder, a granule, a tablet, a granule, a dripping pill, a honey pill, a syrup or an oral liquid; the injection is a solution type, mixed Suspension type, emulsion type or lyophilized powder.
  • the above-mentioned pharmaceutical composition may also be an external dosage form, such as an ointment, gel, spray or patch.
  • the above-mentioned pharmaceutical composition may contain excipients or other pharmaceutically acceptable carriers.
  • the ratio of the auxiliary material to the active ingredient of the medicine may be, for example, the active ingredient accounts for 30% to 50% by weight, and the auxiliary material accounts for 70% to 50% by weight.
  • the auxiliary material can be, for example, one or more of sodium hyaluronate, magnesium stearate, sodium alginate, starch, microcrystalline cellulose, chitosan, stachyose, binder or collagen.
  • Figure 1 shows the chemical structure of ginsenoside Rk3
  • Figure 2 is the chemical structure of ginsenoside Rh4
  • FIG. 3 is a graph showing the results of HE staining of mouse liver tissue in Example 4.
  • FIG. in is a graph showing the results of HE staining of mouse liver tissue in Example 4.
  • the present invention provides a pharmaceutical composition that can be used to prevent and/or treat alcoholic fatty liver, which contains as active ingredients an effective amount of ginsenoside Rk3 and Ginsenoside Rh4 and a pharmaceutically acceptable carrier,
  • the weight ratio of the ginsenoside Rk3 and the ginsenoside Rh4 is 1:0.5-2.
  • ginsenoside Rk3 refers to the compound as shown in Fig. 1, and its molecular formula is C 36 H 60 O 8 .
  • the above-mentioned ginsenoside Rk3 is a known compound, which can be prepared by methods known in the art, for example, by adding an organic acid solution to the triol group ginsenoside under high temperature and high pressure conditions to obtain a higher purity ginsenoside Rk3 after a directional conversion reaction.
  • the crude product is purified by high performance liquid phase separation and other methods to obtain pure ginsenoside Rk3 with a purity of ⁇ 98%.
  • ginsenoside Rh4 refers to the compound shown in Figure 2 and its molecular formula is C 36 H 60 O 8 .
  • the above-mentioned ginsenoside Rh4 is a known compound, which can be prepared by methods known in the art, for example, by adding an organic acid solution of triol group ginsenoside, and undergoing a directional conversion reaction under high temperature and high pressure conditions, and then undergoing separation and purification to obtain a purity of ⁇ 98 % Pure ginsenoside Rh4.
  • the pharmaceutical composition of the present invention may or may not contain other ginsenosides.
  • the content of the ginsenoside Rk3 and ginsenoside Rh4 is preferably 50 parts by weight or more, more preferably 60 parts by weight or more, more preferably 70 parts by weight or more, more preferably 80 parts by weight or more, more preferably 90 parts by weight or more, more preferably 95 parts by weight or more, more preferably 99 parts by weight or more, more preferably 100 parts by weight (That is, the pharmaceutical composition only contains the above two ginsenosides), based on 100 parts by weight of the total ginsenosides contained in the pharmaceutical composition.
  • the purity of ginsenoside Rk3 used in the pharmaceutical composition of the present invention can be more than 98%, and the purity of ginsenoside Rh4 can be more than 98%.
  • the content of the total ginsenosides can be determined by the vanillin method, and the content of the ginsenosides Rk3 and Rh4 can be determined by the HPLC method.
  • the pharmaceutical composition of the present invention may contain other active ingredients for the prevention and/or treatment of alcoholic fatty liver, or may not contain active ingredients for the prevention and/or treatment of alcoholic fatty liver (ie, ginsenoside Rk3 and ginsenoside Rh4 are used as The only active ingredient to prevent and/or treat alcoholic fatty liver).
  • the present invention also provides the use of the pharmaceutical composition of the present invention in the preparation of drugs for the prevention and/or treatment of alcoholic fatty liver.
  • the pharmaceutical composition of the present invention may contain pharmaceutically acceptable carriers, such as excipients.
  • pharmaceutically acceptable carriers such as excipients.
  • excipients there are no special restrictions on the excipients in the pharmaceutical composition of the present invention.
  • the excipients commonly used in medicines or health care products in this technical field can be used.
  • the auxiliary materials are starch, dextrin, lactose, mannitol, sodium hypromellose, xanthan gum, protein sugar and the like.
  • the dosage form of the pharmaceutical composition of the present invention may be an oral dosage form or an injection dosage form.
  • the oral dosage form may be a liquid dosage form or a solid dosage form.
  • the oral dosage form may be, for example, a hard capsule, a soft capsule, a sustained-release capsule, a compressed tablet, a sugar-coated tablet, a powder, a granule, a dripping pill, a honey pill, a syrup, or an oral liquid;
  • the injection dosage form may be, for example, a solution type, Suspension type, emulsion type or lyophilized powder.
  • the administration mode of the pharmaceutical composition for preventing and/or treating alcoholic fatty liver can be, for example, oral, drip or injection.
  • tablets When preparing solid preparations for oral administration, after adding excipients and optional binders, disintegrating agents, lubricants, coloring agents, flavoring agents, etc. to the main drug, tablets can be prepared according to conventional methods. , Coated tablets, granules, fine granules, powders, capsules, etc.
  • lactose corn starch, white sugar, glucose, sorbitol, crystalline cellulose, silicon dioxide, etc.
  • binders for example, polyvinyl alcohol, ethyl cellulose, and methyl cellulose can be used.
  • a lubricant for example, magnesium stearate, talc, silicon dioxide, etc. can be used; as a coloring agent, it can be used and allowed to be added to medicines.
  • the coloring agent as the flavoring agent, cocoa powder, menthol, aromatic acid, peppermint oil, borneol, cinnamon powder can be used.
  • the above-mentioned tablets and granules can also be coated with sugar coating, gelatin coating, and other necessary outer coatings.
  • pH regulators, buffers, suspending aids, solubilizers, stabilizers, isotonic agents, preservatives, etc. can be added to the main drug as needed, and then made into intravenous, subcutaneous, and intramuscular injections according to conventional methods .
  • a freeze-dried product can also be prepared by a conventional method.
  • suspending aid examples include methyl cellulose, Tween 80, hydroxyethyl cellulose, gum arabic, tragacanth powder, sodium carboxymethyl cellulose, polyoxyethylene sorbitol monolaurate, and the like.
  • solubilizer examples include polyoxyethylene hydrogenated castor oil, Tween 80, nicotinamide, polyoxyethylene sorbitol monolaurate, polyethylene glycol, and castor oil fatty acid ethyl ester.
  • examples of stabilizers include sodium sulfite and sodium metasulfite; examples of preservatives include methyl paraben, ethyl paraben, sorbic acid, phenol, cresol, and chlorocresol.
  • the pharmaceutical composition of the present invention is administered to a subject to treat tumors.
  • the subject may be a mammal, for example, a human, a rat, a rabbit, a sheep, a pig, a cow, a cat, a dog, a monkey, etc., preferably a human.
  • the pharmaceutical composition of the present invention can be administered orally or parenterally.
  • the dosage varies depending on the degree of symptoms, patient age, sex, weight, differences in sensitivity, application method, application period, application interval, nature of the pharmaceutical preparation, types of active ingredients, etc., and there is no particular limitation, but usually adults (weight 60Kg)
  • the above application amount can usually be applied 1 to 3 times a day.
  • Example 1 Preparation of tablets with ginsenoside Rk3/Rh4 composition for preventing and treating alcoholic fatty liver
  • ginsenoside Rk3 with a purity of ⁇ 98%
  • 200g of ginsenoside Rh4 with a purity of ⁇ 98%
  • 500g of medicinal excipients commonly used in the preparation of tablets mix them uniformly, press, dry, and package to obtain 2000 tablets of the present invention
  • Each tablet contains 150mg of active ingredient.
  • Example 2 Preparation of capsules with ginsenoside Rk3/Rh4 composition for preventing and treating alcoholic fatty liver
  • ginsenoside Rk3 with a purity of ⁇ 98%
  • 100g of ginsenoside Rh4 with a purity of ⁇ 98%
  • 600g of commonly used pharmaceutical excipients mix them evenly, and then fill them into capsules currently on the market to obtain 2000 capsules of the present invention.
  • the effective ingredient content in each capsule is 100mg.
  • ginsenoside Rk3 with a purity of ⁇ 98% and 80g of ginsenoside Rh4 with a purity of ⁇ 98%, and mix them evenly, then add appropriate amount of flavoring crystalline fructose, stachyose, citric acid, pectin, diluent water, and heat Dissolve, uniformly fill, and sterilize to prepare 1,000 bottles of the oral liquid of the present invention, and each bottle of the oral liquid contains 120 mg of active ingredients.
  • mice 100 adult male mice (18-22 grams) were randomly divided into normal control group; model group; dosage group: group 1 of the composition of the present invention in Example 2 (Rk3 15mg/kg, Rh4 15mg); the present invention Composition 2 groups (Rk3 7.5mg/kg, Rh4 22.5mg), composition 3 groups of the present invention (Rk3 22.5mg/kg, Rh4 7.5mg), ginsenoside Rk3 (30mg/kg) group, ginsenoside Rh4 (30mg/ kg) group, polyene phosphatidylcholine (30mg/kg) positive drug control group, ginsenoside Rg1 (30mg/kg) group, ginsenoside CK/Rh1 (CK 15mg/kg, Rh1 15mg/kg) group, 10 groups Except for the normal control group, mice in the other groups were given 50% ethanol once a day by intragastric administration at a dose of 12 mL/kg BW for 4 consecutive weeks.
  • mice in the other groups were given
  • mice in the normal control group were given an equal volume of purified water by gavage.
  • the test drug was formulated into a suspension with purified water, and was given by intragastric administration once a day.
  • the mice in the normal control group and the model control group were given the same amount of purified water by intragastric administration, and the intragastric volume was 10ml/kg/d.
  • the mice were free to eat and drink, weighed twice a week, and adjusted the amount of test samples according to their body weight.
  • the model control group and each sample group were given 50% ethanol once at the end of the experiment.
  • the mice were given 12ml/kg BW by gavage.
  • the negative control group was given purified water and fasted for 16h.
  • Animals were intraperitoneally injected with 60 mg/kg BW pentobarbital sodium solution anesthetized from the abdominal aorta, and blood was collected from the abdominal aorta, and liver tissues were taken for detection of various indicators: serum ALT, AST and LDL, serum TC, serum TG and LDL content, liver Tissue TG content and histopathological examination.
  • CI ⁇ 1 the two drugs have a synergistic effect, when CI ⁇ 0.7, the synergy is very significant (see David H. Kern, Carol R. Morgan, and Susanne U. Hildebrand-Zanki.
  • the weight difference of each group was not statistically significant (P ⁇ 0.05); compared with the blank group, the liver mass and liver coefficient of the model group were significantly increased (P ⁇ 0.01), compared with the model group, the liver coefficients of each administration group were all Significant decrease (P ⁇ 0.05). Among them, the liver coefficient of Rk3/Rh4 group dropped extremely significantly (P ⁇ 0.01). The results of this test showed that each administration group had a significant inhibitory effect on alcohol-induced liver enlargement in mice.
  • the composition 1 of the present invention has a better effect than each single component, and has a synergistic effect (CI value ⁇ 1), compared with the polyene phosphatidylcholine group, ginsenoside Rg1 and ginsenoside CK/Rh1
  • the liver coefficient reduction effect of the composition 1 of the present invention is more significant.
  • Table 2 The effect of the composition of the present invention on the activity of serum ALT and AST in alcoholic fatty liver mice
  • ALT and AST are the hallmark indicators of liver damage.
  • the content of ALT and AST in normal serum is very low. Only after liver damage, ALT and AST in liver cells are released into the blood by cell rupture, and ethanol is produced during liver metabolism. Reactive oxygen can directly cause oxidative damage to liver cells.
  • Table 2 that the serum ALT and AST activities of animals in each administration group are significantly different from those of the model group (P ⁇ 0.05).
  • the Rk3/Rh4 group of the composition of the present invention has extremely different serum ALT and AST activities from the model group. Significantly (P ⁇ 0.01), very significantly lower than the model group, but not significantly different from the blank control group (P>0.05).
  • composition of the present invention can significantly reduce serum levels of alcoholic fatty liver mice Abnormally elevated ALT and AST activities, the composition 1 of the present invention at the same dose compared to each single component and polyene phosphatidylcholine group, ginsenoside Rg1, ginsenoside CK/Rh1 in the same dose of ALT, AST
  • the activity reduction effect is stronger.
  • the combination index CI of ginsenoside Rk3 and ginsenoside Rh4 composition 1 is ⁇ 1
  • the combination index of composition 2 and composition 3 is CI> 1, indicating that the composition 1 of the present invention is good
  • the synergistic effect of serotonin has a good protective effect on liver cell damage caused by drinking.
  • Table 3 The influence of the composition of the present invention on the serum TC, TG, LDL content and liver tissue TG content of alcoholic fatty liver mice
  • the damage of alcohol to liver cells is achieved through free radical-mediated lipid peroxidation.
  • the composition 1 of the present invention can effectively reduce the content of TC and TG in mouse serum and improve The level of LDL inhibits the synthesis of endogenous TC and TG, and improves the distribution of lipids in the body and the deposition of internal organs.
  • liver of the control group is normal in size, with red color, soft texture, and sharp edges; the liver of the model group is obviously enlarged, gray-white, grainy, and the cut surface is greasy; each The liver morphology of the treatment group was improved compared with the model group, but the liver morphology of the Rk3/Rh4 composition was closer to that of the control group than the other treatment groups of R.
  • the results of HE staining of the liver ( Figure 3, 1-10) showed that the liver cells in the control group were normal in size and morphology, neatly arranged, with a large and round nucleus visible in the center, and the cytoplasm was evenly colored.
  • lipid droplets were deposited in the liver cells, and the morphology and structure were normal, without steatosis and inflammation; the liver cells in the model group were swollen and the boundaries were unclear.
  • the fatty degeneration cells are distributed in a focal shape.
  • the liver tissue is almost replaced by fatty vacuoles.
  • the hepatocytes are balloon-like degeneration, and there are inflammatory cell infiltration in the portal area; human saponin Rk3 , Rh4 single action group, composition 2 group, composition 3 group, positive drug control group, ginsenoside Rg1 group, and ginsenoside CK/Rh1 group.
  • mice fed with the Rk3/Rh4 composition 1 of the present invention had almost no lipid droplet deposition in the liver cells in the liver tissue, the morphology and structure were close to normal, and there was no steatosis.
  • the disclosed Rk3/Rh4 composition can better improve the degree of liver steatosis in mice caused by alcohol.
  • Example 4 The normal control group in Example 4; model group; Rk3 group, Rh4 group, Rk3/Rh4 group in liver superoxide dismutase (SOD), liver reduced glutathione (GSH), and liver malondioxide Aldehyde (MDA) level; and molecular chip screening to determine the expression level of related genes in hepatocytes, and the test results are based on the mean plus or minus the standard deviation Indicates that the statistical processing method uses one-way analysis of variance;
  • the liver cells of alcoholic fatty liver are very susceptible to oxidative stress.
  • the model group mice have weakened antioxidant capacity, which is manifested by increased levels of MDA, a harmful product of alcohol metabolism, decreased liver tissue SOD activity, and decreased levels of reduced glutathione GSH.
  • MDA manganese-associated dihydroxyase
  • GSH glutathione
  • composition of the present invention has a statistical table showing the relative expression of genes related to important metabolic pathways in alcoholic fatty liver mice
  • ⁇ the corresponding gene is down-regulated fold in liver cell mRNA expression
  • ⁇ indicates the corresponding gene is up-regulated fold in liver cell mRNA expression.
  • Fatty acid synthase which plays a vital role in fatty acid synthesis, is responsible for all the catalytic steps for the de novo synthesis of acetyl-CoA and malonyl-CoA to a long-chain palmitate (palmitate).
  • This gene The expression directly affects the amount of fatty acid synthase and plays an important role in controlling animal fat deposition.
  • Lipoprotein lipase (LpL) is one of the key enzymes of fat metabolism. It acts on lipoproteins and mainly decomposes triacylglycerols in lipoproteins. It is the rate-limiting enzyme for removing triacylglycerols in plasma and promotes the transfer of cholesterol from lipoproteins.
  • Scd1 Sterol-CoA desaturase 1
  • LpL Phospholipids and apolipoprotein
  • LpL also has the ability to increase the ability of chylomicrons to bind to LP receptors, and promote the uptake of chylomicrons.
  • Sterol-CoA desaturase 1 (Scd1) is the rate-limiting enzyme of monounsaturated fatty acid biosynthesis. It plays a central regulatory role in fatty acid metabolism and is also one of the target genes of leptin; leptin and diabetes , Obesity and fatty liver are closely related and are one of the hotspots of metabolic diseases in recent years.
  • the model group can increase the synthesis of Fasn and decrease the synthesis of LpL, which is the culprit leading to excessive fat deposition in liver cells and the primary factor in the occurrence of fatty liver; and each group of administration can reduce the synthesis of Fasn, the administration group Rh4 and the composition Rk3/Rh4 can significantly increase LpL synthesis, reduce fatty acid synthesis in liver tissue, strengthen lipolysis, and have a positive effect on alleviating liver fatness.
  • the up-regulation of Scd1 gene expression in the model group may be one of the most important molecular mechanisms that cause alcoholic steatosis, but each administration group can significantly down-regulate the expression of Scd1 gene.
  • composition of the present invention has the effect of preventing and treating alcoholic fatty liver, and it has no side effects after long-term use.
  • the present invention provides a pharmaceutical composition containing ginsenosides Rk3 and Rh4, which is synergistic and has the effect of preventing and/or treating alcoholic fatty liver.

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Abstract

Disclosed in the present invention is a ginsenoside composition having an alcoholic fatty liver-preventing and -treating function. The pharmaceutical composition able to be used to prevent and/or treat alcoholic fatty liver of the present invention contains a prophylactically and/or therapeutically effective dose of ginsenoside Rk3 and ginsenoside Rh4 as active ingredients, and a pharmaceutically acceptable carrier. In the pharmaceutical composition, the mass ratio between the ginsenoside Rk3 and the ginsenoside Rh4 is 1:0.5-2, and is preferably 1:0.9-1.1. The ginsenoside composition of the present invention is able to markedly improve liver function index abnormalities caused by a large amount of alcohol consumption, reducing the generation of inflammatory infiltrates and fat vacuoles in liver tissue. The present invention provides a synergistically interacting pharmaceutical composition containing ginsenosides Rk3 and Rh4 and having alcoholic fatty liver-preventing and/or -treating effects.

Description

一种具有防治酒精性脂肪肝功能的人参皂苷组合物Ginsenoside composition with function of preventing and treating alcoholic fatty liver 技术领域Technical field
本发明涉及生物医药领域，具体涉及一种具有防治酒精性脂肪肝的人参皂苷Rk3/Rh4组合物。The invention relates to the field of biomedicine, in particular to a ginsenoside Rk3/Rh4 composition capable of preventing and treating alcoholic fatty liver.
背景技术Background technique
脂肪性肝病是一种常见病，根据有无大量饮酒史可分为酒精性脂肪性肝病(alcoholic fatty liver，AFL)与非酒精性脂肪肝(NAFLD)，AFL是由于长期饮用酒精导致酒精性肝损伤，其主要原因是酒精在肝细胞内代谢产生的毒性代谢产物、氧化应激及其引起的代谢紊乱。而NAFLD的发病机制主要是与IR、肝脏脂肪代谢异常、线粒体功能障碍和氧化应激，遗传变异和代谢改变及细胞损伤的易感性等相关。尽管病毒性肝炎所致的各种肝病占主要地位，但随着酒精在人类饮食结构的地位上升，酒精性脂肪肝的病发率越来越高，其发生肝硬化和肝癌的机会要远超过NAFLD。成为了继病毒性肝炎后导致肝损害的第二大病因。Fatty liver disease is a common disease. It can be divided into alcoholic fatty liver (AFL) and non-alcoholic fatty liver (NAFLD) according to the history of heavy drinking. AFL is caused by long-term alcohol consumption. The main causes of injury are toxic metabolites, oxidative stress and metabolic disorders caused by alcohol metabolism in liver cells. The pathogenesis of NAFLD is mainly related to IR, abnormal liver fat metabolism, mitochondrial dysfunction and oxidative stress, genetic variation and metabolic changes, and susceptibility to cell damage. Although various liver diseases caused by viral hepatitis occupy a major position, with the rise of alcohol in the human diet, the incidence of alcoholic fatty liver is getting higher and higher, and the chance of developing liver cirrhosis and liver cancer far exceeds NAFLD. It has become the second leading cause of liver damage after viral hepatitis.
多烯磷脂酰胆碱是辅助治疗酒精性脂肪肝的常用药，能有效减少氧自由基，保护肝细胞膜，延缓肝纤维化，从而促患者恢复，但临床认为单药治疗，患者恢复较慢，容易引起肠道紊乱从而导致腹泻。Polyene phosphatidylcholine is a commonly used drug for adjuvant treatment of alcoholic fatty liver. It can effectively reduce oxygen free radicals, protect liver cell membranes, and delay liver fibrosis, thereby promoting the recovery of patients. Easily cause intestinal disorders and cause diarrhea.
人参皂苷是人参中的活性成分，是一种固醇类化合物，能影响机体内的多重代谢通路。有研究报道，人参皂苷Rk3、Rh4具有明显的抗肿瘤、抗氧化、抗炎、抑制细胞凋亡等作用，也有文献报道人参皂苷CK、Rh1及其组合物能够应用于改善非酒精性脂肪性肝纤维化与胰岛素抵抗药物中，但未对酒精性脂肪肝进行研究；人参皂苷Rg1在治疗饮酒导致代谢紊乱方面有一定功效，但对肝脏是否有保护作用并未研究过，而人参皂苷Rk3/Rh4联合用药从未运用于改善因酒精所引起的脂肪肝、肝病以及肝脏抗氧化物质减少等。Ginsenoside is the active ingredient in ginseng and a sterol compound that can affect multiple metabolic pathways in the body. Studies have reported that ginsenosides Rk3 and Rh4 have obvious anti-tumor, anti-oxidant, anti-inflammatory, and anti-apoptosis effects. There are also reports in the literature that ginsenosides CK, Rh1 and their combinations can be used to improve non-alcoholic fatty liver In fibrosis and insulin resistance drugs, alcoholic fatty liver has not been studied; ginsenoside Rg1 has a certain effect in the treatment of metabolic disorders caused by drinking, but whether it has a protective effect on the liver has not been studied, and ginsenoside Rk3/Rh4 Combination drugs have never been used to improve fatty liver, liver disease and the reduction of liver antioxidants caused by alcohol.
发明内容Summary of the invention
本发明的目的在于提供具有防治酒精性脂肪肝功能的人参皂苷Rk3/Rh4组合物。The purpose of the present invention is to provide a ginsenoside Rk3/Rh4 composition with the function of preventing and treating alcoholic fatty liver.
发明人发现人参皂苷Rk3/Rh4联用能有效改善饮酒导致的肝脏损伤和脂肪变性程度，并由此将人参皂苷Rk3/Rh4组合物进一步应用到了酒精性脂肪肝的治疗、缓解方面，且该组合物是首次将人参皂苷Rk3/Rh4组合物为主要成分应用于防治酒精性脂肪肝方面，该组合物对酒精性脂肪肝具有良好的治疗、改善效果。在小鼠酒精性脂肪肝模型中，Rk3/Rh4组合物可显著提高饮酒小鼠生存率，降低血清内谷丙转氨酶(ALT)和谷草转氨酶含量(AST)， 减少血清及肝脏中胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)的含量，减少饮酒导致的肝脏中脂质堆积(油滴)、肝脏组织炎性浸润和脂肪空泡。The inventor found that the combination of ginsenoside Rk3/Rh4 can effectively improve the degree of liver damage and steatosis caused by drinking, and thus further applied the ginsenoside Rk3/Rh4 composition to the treatment and alleviation of alcoholic fatty liver, and the combination This is the first time that the ginsenoside Rk3/Rh4 composition is used as the main component in the prevention and treatment of alcoholic fatty liver. The composition has good therapeutic and improving effects on alcoholic fatty liver. In a mouse alcoholic fatty liver model, the Rk3/Rh4 composition can significantly improve the survival rate of drinking mice, reduce serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and reduce serum and liver cholesterol (TC) , Triglyceride (TG), low-density lipoprotein (LDL) content, reduce lipid accumulation (oil droplets) in the liver caused by drinking, liver tissue inflammatory infiltration and fatty vacuoles.
即，本发明包括：That is, the present invention includes:
1.一种可用于预防和/或治疗酒精性脂肪肝的药物组合物，其包含作为活性成分的预防和/或治疗有效量的人参皂苷Rk3和人参皂苷Rh4、以及药学可接受的载体，1. A pharmaceutical composition that can be used for the prevention and/or treatment of alcoholic fatty liver, which comprises preventive and/or therapeutically effective amounts of ginsenoside Rk3 and ginsenoside Rh4 as active ingredients, and a pharmaceutically acceptable carrier,
该药物组合物中，所述人参皂苷Rk3和人参皂苷Rh4的重量比为1:0.5～2、优选1:0.9～1.1。In the pharmaceutical composition, the weight ratio of ginsenoside Rk3 and ginsenoside Rh4 is 1:0.5-2, preferably 1:0.9-1.1.
2.根据项1所述的药物组合物，所述药物组合物中，所述人参皂苷Rk3和人参皂苷Rh4的重量比为1：1。2. The pharmaceutical composition according to item 1, in the pharmaceutical composition, the weight ratio of ginsenoside Rk3 and ginsenoside Rh4 is 1:1.
3.根据项1所述的药物组合物，以所述药物组合物中包含的总人参皂苷为100重量份计，所述药物组合物中包含的人参皂苷Rk3和人参皂苷Rh4合计为10重量份以上。3. The pharmaceutical composition according to item 1, based on 100 parts by weight of total ginsenosides contained in the pharmaceutical composition, the total of ginsenoside Rk3 and ginsenoside Rh4 contained in the pharmaceutical composition is 10 parts by weight above.
4.根据项1所述的药物组合物，以所述药物组合物中包含的总人参皂苷为100重量份计，所述药物组合物中包含的人参皂苷Rk3和人参皂苷Rh4合计为100重量份。4. The pharmaceutical composition according to item 1, based on 100 parts by weight of total ginsenosides contained in the pharmaceutical composition, the total of ginsenoside Rk3 and ginsenoside Rh4 contained in the pharmaceutical composition is 100 parts by weight .
5.根据项1所述的用途，其中，所述药物组合物不包含除人参皂苷Rk3和人参皂苷Rh4之外的其它预防和/或治疗酒精性脂肪肝的有效成分。5. The use according to item 1, wherein the pharmaceutical composition does not contain other effective ingredients for preventing and/or treating alcoholic fatty liver except ginsenoside Rk3 and ginsenoside Rh4.
6.人参皂苷Rk3和人参皂苷Rh4在制备用于预防和/或治疗酒精性脂肪肝的药物组合物中的用途，其中，该药物组合物中，所述人参皂苷Rk3和人参皂苷Rh4的重量比为1:0.5～2、优选1:0.9～1.1。6. The use of ginsenoside Rk3 and ginsenoside Rh4 in the preparation of a pharmaceutical composition for the prevention and/or treatment of alcoholic fatty liver, wherein the weight ratio of the ginsenoside Rk3 and ginsenoside Rh4 in the pharmaceutical composition It is 1:0.5-2, preferably 1:0.9-1.1.
7.根据项6所述的用途，其中，所述药物组合物中，所述人参皂苷Rk3和人参皂苷Rh4的重量比为1：1。7. The use according to item 6, wherein, in the pharmaceutical composition, the weight ratio of ginsenoside Rk3 and ginsenoside Rh4 is 1:1.
8.根据项6所述的用途，其中，以所述药物组合物中包含的总人参皂苷为100重量份计，所述药物组合物中包含的人参皂苷Rk3和人参皂苷Rh4合计为10重量份以上。8. The use according to item 6, wherein, based on 100 parts by weight of the total ginsenosides contained in the pharmaceutical composition, the total of ginsenoside Rk3 and ginsenoside Rh4 contained in the pharmaceutical composition is 10 parts by weight above.
9.根据项6所述的用途，以所述药物组合物中包含的总人参皂苷为100重量份计，所述药物组合物中包含的人参皂苷Rk3和人参皂苷Rh4合计为100重量份。9. The use according to item 6, based on 100 parts by weight of total ginsenosides contained in the pharmaceutical composition, the total of ginsenoside Rk3 and ginsenoside Rh4 contained in the pharmaceutical composition is 100 parts by weight.
10.根据项6所述的用途，其中，所述药物组合物不包含除人参皂苷Rk3和人参皂苷Rh4之外的其它预防和/或治疗酒精性脂肪肝的有效成分。10. The use according to item 6, wherein the pharmaceutical composition does not contain other effective ingredients for preventing and/or treating alcoholic fatty liver except ginsenoside Rk3 and ginsenoside Rh4.
本发明所述的防治酒精性脂肪肝的人参皂苷Rk3/Rh4组合物，既可以是药物，也可以是其他任何制剂形式；既可以是治疗相关疾病为目的，也可以是改善或预防饮酒导致的脂肪肝。该制剂可以是人参皂苷Rk3/Rh4，也可以是人参皂苷Rk3/Rh4和其它成分组成的复方制剂，其中人参皂苷Rk3/Rh4作为主要成分。The ginsenoside Rk3/Rh4 composition for the prevention and treatment of alcoholic fatty liver of the present invention can be either a medicine or any other preparation form; it can be used for the purpose of treating related diseases, or it can be used to improve or prevent drinking caused by alcohol. Fatty liver. The preparation can be ginsenoside Rk3/Rh4, or a compound preparation composed of ginsenoside Rk3/Rh4 and other components, wherein ginsenoside Rk3/Rh4 is the main component.
本发明所述人参皂苷Rk3/Rh4可以由人参或者西洋参等提取转化分离而得，也可以是通过化学合成而得。具体来讲，所述人参皂苷Rk3/Rh4可 由人参或西洋参的根、茎和/或叶，利用现有技术的工艺方法提取转化分离而得。The ginsenoside Rk3/Rh4 of the present invention can be obtained by extraction, transformation and separation of ginseng or American ginseng, or it can be obtained by chemical synthesis. Specifically, the ginsenoside Rk3/Rh4 can be obtained from the roots, stems, and/or leaves of ginseng or American ginseng by extracting, transforming, and isolating them using prior art processes.
此外，发明人还发现，在小鼠酒精性脂肪肝模型中，Rk3或Rh4可提高饮酒小鼠生存率，降低血清内谷丙转氨酶(ALT)和谷草转氨酶含量(AST)，减少血清及肝脏中胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)的含量，减少饮酒导致的肝脏中脂质堆积(油滴)、肝脏组织炎性浸润和脂肪空泡。In addition, the inventor also found that in a mouse alcoholic fatty liver model, Rk3 or Rh4 can increase the survival rate of drinking mice, reduce serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and reduce serum and liver The content of cholesterol (TC), triglycerides (TG), and low-density lipoprotein (LDL) reduces the accumulation of lipids (oil droplets) in the liver, inflammatory infiltration of liver tissue and fatty vacuoles caused by drinking.
因此，本发明还包括：Therefore, the present invention also includes:
11.人参皂苷Rk3在制备用于预防和/或治疗酒精性脂肪肝的药物组合物中的用途。11. Use of ginsenoside Rk3 in the preparation of a pharmaceutical composition for preventing and/or treating alcoholic fatty liver.
12.根据项11所述的用途，其中，以所述药物组合物中包含的总人参皂苷为100重量份计，所述药物组合物中包含的人参皂苷Rk3为5重量份以上、优选10重量份以上、更优选20重量份以上、更优选50重量份以上、更优选70重量份以上、更优选80重量份以上、更优选90重量份以上、更优选95重量份以上、更优选99重量份以上、更优选为100重量份。12. The use according to item 11, wherein, based on 100 parts by weight of the total ginsenosides contained in the pharmaceutical composition, the ginsenoside Rk3 contained in the pharmaceutical composition is 5 parts by weight or more, preferably 10 parts by weight Parts by weight or more, more preferably 20 parts by weight or more, more preferably 50 parts by weight or more, more preferably 70 parts by weight or more, more preferably 80 parts by weight or more, more preferably 90 parts by weight or more, more preferably 95 parts by weight or more, more preferably 99 parts by weight Above, more preferably 100 parts by weight.
13.根据项11所述的用途，其中，所述药物组合物不包含除人参皂苷Rk3之外的其它预防和/或治疗酒精性脂肪肝的有效成分。13. The use according to item 11, wherein the pharmaceutical composition does not contain other effective ingredients for preventing and/or treating alcoholic fatty liver other than ginsenoside Rk3.
14.人参皂苷Rh4在制备用于预防和/或治疗酒精性脂肪肝的药物组合物中的用途。14. Use of ginsenoside Rh4 in the preparation of a pharmaceutical composition for the prevention and/or treatment of alcoholic fatty liver.
15.根据项14所述的用途，其中，以所述药物组合物中包含的总人参皂苷为100重量份计，所述药物组合物中包含的人参皂苷Rh4为5重量份以上、优选10重量份以上、更优选20重量份以上、更优选50重量份以上、更优选70重量份以上、更优选80重量份以上、更优选90重量份以上、更优选95重量份以上、更优选99重量份以上、更优选为100重量份。15. The use according to item 14, wherein, based on 100 parts by weight of the total ginsenosides contained in the pharmaceutical composition, the ginsenoside Rh4 contained in the pharmaceutical composition is 5 parts by weight or more, preferably 10 parts by weight Parts or more, more preferably 20 parts by weight or more, more preferably 50 parts by weight or more, more preferably 70 parts by weight or more, more preferably 80 parts by weight or more, more preferably 90 parts by weight or more, more preferably 95 parts by weight or more, more preferably 99 parts by weight Above, more preferably 100 parts by weight.
16.根据项14所述的用途，其中，所述药物组合物不包含除人参皂苷Rh4之外的其它预防和/或治疗酒精性脂肪肝的有效成分。16. The use according to item 14, wherein the pharmaceutical composition does not contain other effective ingredients for preventing and/or treating alcoholic fatty liver other than ginsenoside Rh4.
上述药物组合物可以是例如口服剂或者注射剂。所述口服剂可以是例如硬胶囊、软胶囊、缓控释放胶囊、糖衣片、粉剂、颗粒剂、片剂、冲剂、滴丸、水蜜丸、糖浆或口服液；所述注射剂为溶液型、混悬液型、乳浊液型或冻干粉。上述药物组合物还可以是外用剂型，例如膏剂、凝胶剂、喷剂或贴膏。上述药物组合物中可包含辅料或其它药学可接受的载体。所述辅料与药物有效成分之比可以是例如，有效成分占30重量％～50重量％，辅料占70重量％～50重量％。所述辅料可以为例如透明质酸钠、硬脂酸镁、海藻酸钠、淀粉、微晶纤维素、壳聚糖、水苏糖、粘合剂或者胶原蛋白中的一种或几种。The above-mentioned pharmaceutical composition may be, for example, an oral agent or an injection. The oral agent may be, for example, a hard capsule, a soft capsule, a sustained-release capsule, a sugar-coated tablet, a powder, a granule, a tablet, a granule, a dripping pill, a honey pill, a syrup or an oral liquid; the injection is a solution type, mixed Suspension type, emulsion type or lyophilized powder. The above-mentioned pharmaceutical composition may also be an external dosage form, such as an ointment, gel, spray or patch. The above-mentioned pharmaceutical composition may contain excipients or other pharmaceutically acceptable carriers. The ratio of the auxiliary material to the active ingredient of the medicine may be, for example, the active ingredient accounts for 30% to 50% by weight, and the auxiliary material accounts for 70% to 50% by weight. The auxiliary material can be, for example, one or more of sodium hyaluronate, magnesium stearate, sodium alginate, starch, microcrystalline cellulose, chitosan, stachyose, binder or collagen.
附图说明Description of the drawings
图1为人参皂苷Rk3化学结构式Figure 1 shows the chemical structure of ginsenoside Rk3
图2为人参皂苷Rh4化学结构式Figure 2 is the chemical structure of ginsenoside Rh4
图3为显示实施例4中的小鼠肝脏组织HE染色结果的图。其中，FIG. 3 is a graph showing the results of HE staining of mouse liver tissue in Example 4. FIG. in,
1：对照组小鼠的肝脏组织HE染色图；1: HE staining image of liver tissue of mice in the control group;
2：模型组小鼠的肝脏组织HE染色图；2: HE staining image of liver tissue of mice in the model group;
3：阳性药对照组小鼠的肝脏组织HE染色图；3: HE staining image of liver tissue of mice in the positive drug control group;
4：人参皂苷Rk3组小鼠的肝脏组织HE染色图；4: HE staining image of liver tissue of mice in ginsenoside Rk3 group;
5：人参皂苷Rh4组小鼠的肝脏组织HE染色图；5: HE staining image of liver tissue of mice in the ginsenoside Rh4 group;
6：人参皂苷Rk3/Rh4组合物1组小鼠的肝脏组织HE染色图；6: HE staining image of liver tissue of mice with ginsenoside Rk3/Rh4 composition 1 group;
7：人参皂苷Rk3/Rh4组合物2组小鼠的肝脏组织HE染色图；7: HE staining image of liver tissue of mice with ginsenoside Rk3/Rh4 composition 2 group;
8：人参皂苷Rk3/Rh4组合物3组小鼠的肝脏组织HE染色图；8: HE staining image of liver tissue of mice with ginsenoside Rk3/Rh4 composition 3 groups;
9：人参皂苷Rg1组小鼠的肝脏组织HE染色图；9: HE staining image of liver tissue of mice in ginsenoside Rg1 group;
10：人参皂苷CK/Rh1组小鼠的肝脏组织HE染色图；10: HE staining image of liver tissue of mice in ginsenoside CK/Rh1 group;
发明的具体实施方式Specific embodiments of the invention
首先，在一个方面中，本发明提供一种可用于预防和/或治疗酒精性脂肪肝的药物组合物，其包含作为活性成分的预防和/或治疗酒精性脂肪肝有效量的人参皂苷Rk3和人参皂苷Rh4、以及药学可接受的载体，First, in one aspect, the present invention provides a pharmaceutical composition that can be used to prevent and/or treat alcoholic fatty liver, which contains as active ingredients an effective amount of ginsenoside Rk3 and Ginsenoside Rh4 and a pharmaceutically acceptable carrier,
该药物组合物中，所述人参皂苷Rk3和人参皂苷Rh4的重量比为1:0.5～2。In the pharmaceutical composition, the weight ratio of the ginsenoside Rk3 and the ginsenoside Rh4 is 1:0.5-2.
在本说明书中，人参皂苷Rk3是指如图1所示的化合物，其分子式为C 36H 60O 8。 In this specification, ginsenoside Rk3 refers to the compound as shown in Fig. 1, and its molecular formula is C 36 H 60 O 8 .
上述人参皂苷Rk3是已知化合物，可以采用本技术领域已知的方法来制备，例如通过在三醇组人参皂苷中加有机酸溶液高温高压条件下定向转化反应后获得较高纯度的人参皂苷Rk3粗品，再经过高效液相分离等方法纯化后得到纯度≥98％的人参皂苷Rk3纯品。The above-mentioned ginsenoside Rk3 is a known compound, which can be prepared by methods known in the art, for example, by adding an organic acid solution to the triol group ginsenoside under high temperature and high pressure conditions to obtain a higher purity ginsenoside Rk3 after a directional conversion reaction. The crude product is purified by high performance liquid phase separation and other methods to obtain pure ginsenoside Rk3 with a purity of ≥98%.
在本说明书中，人参皂苷Rh4指如图2所示的化合物，其分子式为C 36H 60O 8。 In this specification, ginsenoside Rh4 refers to the compound shown in Figure 2 and its molecular formula is C 36 H 60 O 8 .
上述人参皂苷Rh4是已知化合物，可以采用本技术领域已知的方法来制备，例如通过三醇组人参皂苷加有机酸溶液，高温高压条件下定向转化反应后，再经过分离纯化得到纯度≥98％的人参皂苷Rh4纯品。The above-mentioned ginsenoside Rh4 is a known compound, which can be prepared by methods known in the art, for example, by adding an organic acid solution of triol group ginsenoside, and undergoing a directional conversion reaction under high temperature and high pressure conditions, and then undergoing separation and purification to obtain a purity of ≥98 % Pure ginsenoside Rh4.
发明人发现，将人参皂苷Rk3、人参皂苷Rh4按本发明限定的比例范围进行组合，在预防和/或治疗酒精性脂肪肝方面表现出协同增效(CI值小于1、优选CI值小于0.8、更优选小于0.7)。发明人还发现，在上述比例范围内，人参皂苷Rk3和人参皂苷Rh4也是低毒性的。而且，发明人发现，如果所述药物组合物中人参皂苷Rk3、人参皂苷Rh4的含量不在本发明限定的比例范围内，则不具有协同增效作用。从协同增效作用显著的角度考虑，所述组合物中，所述人参皂苷Rk3和人参皂苷Rh4的重量比为优选为1:0.9～1.1，更优选为1:1。The inventor found that the combination of ginsenoside Rk3 and ginsenoside Rh4 in the ratio range defined by the present invention showed synergistic effects in the prevention and/or treatment of alcoholic fatty liver (CI value less than 1, preferably CI value less than 0.8, More preferably less than 0.7). The inventor also found that within the above-mentioned ratio range, ginsenoside Rk3 and ginsenoside Rh4 are also low-toxic. Moreover, the inventor found that if the content of ginsenoside Rk3 and ginsenoside Rh4 in the pharmaceutical composition is not within the ratio range defined by the present invention, there is no synergistic effect. From the perspective of a significant synergistic effect, the weight ratio of the ginsenoside Rk3 and ginsenoside Rh4 in the composition is preferably 1:0.9 to 1.1, and more preferably 1:1.
本发明的药物组合物中，可以包含其它人参皂苷，也可以不包含其它人参皂苷。在本发明的药物组合物中包含其它人参皂苷的情况下，从更好地发挥协同增效作用的角度考虑，所述人参皂苷Rk3和人参皂苷Rh4的含量优 选为50重量份以上、更优选为60重量份以上、更优选为70重量份以上、更优选为80重量份以上、更优选为90重量份以上、更优选为95重量份以上、更优选为99重量份以上、更优选100重量份(即所述药物组合物中仅包含上述两种人参皂苷)，以该药物组合物中包含的总人参皂苷为100重量份计。本发明的药物组合物使用的人参皂苷Rk3纯度可以为98％以上，人参皂苷Rh4纯度可以为98％以上。The pharmaceutical composition of the present invention may or may not contain other ginsenosides. In the case where the pharmaceutical composition of the present invention contains other ginsenosides, from the perspective of better exerting a synergistic effect, the content of the ginsenoside Rk3 and ginsenoside Rh4 is preferably 50 parts by weight or more, more preferably 60 parts by weight or more, more preferably 70 parts by weight or more, more preferably 80 parts by weight or more, more preferably 90 parts by weight or more, more preferably 95 parts by weight or more, more preferably 99 parts by weight or more, more preferably 100 parts by weight (That is, the pharmaceutical composition only contains the above two ginsenosides), based on 100 parts by weight of the total ginsenosides contained in the pharmaceutical composition. The purity of ginsenoside Rk3 used in the pharmaceutical composition of the present invention can be more than 98%, and the purity of ginsenoside Rh4 can be more than 98%.
所述总人参皂苷的含量可以采用香草醛法测定，所述人参皂苷Rk3、Rh4的含量均可以采用HPLC法测定。The content of the total ginsenosides can be determined by the vanillin method, and the content of the ginsenosides Rk3 and Rh4 can be determined by the HPLC method.
本发明的药物组合物中，可以包含其它预防和/或治疗酒精性脂肪肝活性成分，也可以不包含预防和/或治疗酒精性脂肪肝活性成分(即，以人参皂苷Rk3、人参皂苷Rh4作为唯一预防和/或治疗酒精性脂肪肝活性成分)。The pharmaceutical composition of the present invention may contain other active ingredients for the prevention and/or treatment of alcoholic fatty liver, or may not contain active ingredients for the prevention and/or treatment of alcoholic fatty liver (ie, ginsenoside Rk3 and ginsenoside Rh4 are used as The only active ingredient to prevent and/or treat alcoholic fatty liver).
另一方面，发明人发现本发明的药物组合物具有显著的预防和/或治疗酒精性脂肪肝效果。因此，本发明也提供本发明的药物组合物在制备预防和/或治疗酒精性脂肪肝药物中的用途。On the other hand, the inventor found that the pharmaceutical composition of the present invention has a significant preventive and/or therapeutic effect of alcoholic fatty liver. Therefore, the present invention also provides the use of the pharmaceutical composition of the present invention in the preparation of drugs for the prevention and/or treatment of alcoholic fatty liver.
本发明的药物组合物中可以包含药物可接受的载体，例如辅料。对于本发明的药物组合物中的辅料，没有特殊限制，例如可以使用本技术领域通常用于药品或保健品的辅料。具体地，所述辅料为淀粉、糊精、乳糖、甘露醇、羟丙甲纤维素钠、黄原胶、蛋白糖等。The pharmaceutical composition of the present invention may contain pharmaceutically acceptable carriers, such as excipients. There are no special restrictions on the excipients in the pharmaceutical composition of the present invention. For example, the excipients commonly used in medicines or health care products in this technical field can be used. Specifically, the auxiliary materials are starch, dextrin, lactose, mannitol, sodium hypromellose, xanthan gum, protein sugar and the like.
对本发明的药物组合物的剂型没有特殊限制，例如可以是口服剂型或注射剂型。所述口服剂型可以是液体剂型，也可以是固体剂型。所述口服剂型可以为例如硬胶囊、软胶囊、缓控释放胶囊、压片、糖衣片、粉剂、颗粒剂、滴丸、水蜜丸、糖浆或口服液；所述注射剂型可以为例如溶液型、混悬液型、乳浊液型或冻干粉。所述用于预防和/或治疗酒精性脂肪肝的药物组合物的给药方式可以为例如口服、滴注或注射。There is no particular limitation on the dosage form of the pharmaceutical composition of the present invention, for example, it may be an oral dosage form or an injection dosage form. The oral dosage form may be a liquid dosage form or a solid dosage form. The oral dosage form may be, for example, a hard capsule, a soft capsule, a sustained-release capsule, a compressed tablet, a sugar-coated tablet, a powder, a granule, a dripping pill, a honey pill, a syrup, or an oral liquid; the injection dosage form may be, for example, a solution type, Suspension type, emulsion type or lyophilized powder. The administration mode of the pharmaceutical composition for preventing and/or treating alcoholic fatty liver can be, for example, oral, drip or injection.
制备口服用固体制剂时，可以在向主药中加入赋形剂以及视需要而定的粘合剂、崩解剂、滑润剂、着色剂、矫味剂等后，按照常规方法制成片剂、包衣片剂、颗粒剂、细粒剂、散剂、胶囊剂等。When preparing solid preparations for oral administration, after adding excipients and optional binders, disintegrating agents, lubricants, coloring agents, flavoring agents, etc. to the main drug, tablets can be prepared according to conventional methods. , Coated tablets, granules, fine granules, powders, capsules, etc.
作为赋形剂，可使用例如乳糖、玉米淀粉、白糖、葡萄糖、山梨糖醇、结晶纤维素、二氧化硅等；作为粘合剂，可使用例如聚乙烯醇、乙基纤维素、甲基纤维素、阿拉伯胶、羟基丙基纤维素、羟基丙基甲基纤维素等；作为滑润剂，可使用例如硬脂酸镁、滑石、二氧化硅等；作为着色剂，可使用允许在药品中添加的着色剂；作为矫味剂，可使用可可粉、薄荷脑、芳香酸、薄荷油、龙脑、桂皮粉。当然，也可以在上述片剂、颗粒剂上包覆糖衣、明胶衣、以及其它的必要外衣。As excipients, for example, lactose, corn starch, white sugar, glucose, sorbitol, crystalline cellulose, silicon dioxide, etc. can be used; as binders, for example, polyvinyl alcohol, ethyl cellulose, and methyl cellulose can be used. As a lubricant, for example, magnesium stearate, talc, silicon dioxide, etc. can be used; as a coloring agent, it can be used and allowed to be added to medicines. The coloring agent; as the flavoring agent, cocoa powder, menthol, aromatic acid, peppermint oil, borneol, cinnamon powder can be used. Of course, the above-mentioned tablets and granules can also be coated with sugar coating, gelatin coating, and other necessary outer coatings.
制备注射剂时，可以根据需要向主药中添加pH调节剂、缓冲剂、悬助剂、增溶剂、稳定剂、等渗剂、防腐剂等，再按照常规方法制成静脉、皮下、肌肉内注射剂。此时，也可以根据需要，利用常规方法制成冷冻干燥物。When preparing injections, pH regulators, buffers, suspending aids, solubilizers, stabilizers, isotonic agents, preservatives, etc. can be added to the main drug as needed, and then made into intravenous, subcutaneous, and intramuscular injections according to conventional methods . At this time, if necessary, a freeze-dried product can also be prepared by a conventional method.
作为悬助剂，可列举例如甲基纤维素、吐温80、羟基乙基纤维素、阿拉伯胶、黄蓍树胶粉、羧甲基纤维素钠、聚氧乙烯山梨糖醇单月桂酸盐等。Examples of the suspending aid include methyl cellulose, Tween 80, hydroxyethyl cellulose, gum arabic, tragacanth powder, sodium carboxymethyl cellulose, polyoxyethylene sorbitol monolaurate, and the like.
作为增溶剂，可列举例如聚氧化乙烯氢化蓖麻油、吐温80、烟酰胺、聚氧乙烯山梨糖醇单月桂酸盐、聚乙二醇、蓖麻油脂肪酸乙酯等。Examples of the solubilizer include polyoxyethylene hydrogenated castor oil, Tween 80, nicotinamide, polyoxyethylene sorbitol monolaurate, polyethylene glycol, and castor oil fatty acid ethyl ester.
另外，作为稳定剂，可列举例如亚硫酸钠、偏亚硫酸钠等；作为防腐剂，可列举例如对羟基苯甲酸甲酯、对羟基苯甲酸乙酯、山梨酸、苯酚、甲酚、氯甲酚等。In addition, examples of stabilizers include sodium sulfite and sodium metasulfite; examples of preservatives include methyl paraben, ethyl paraben, sorbic acid, phenol, cresol, and chlorocresol.
将本发明的药物组合物施用于对象，可以治疗的肿瘤。所述对象可以是哺乳动物，例如可以是人、大鼠、兔、羊、猪、牛、猫、狗、猴等，优选为人。The pharmaceutical composition of the present invention is administered to a subject to treat tumors. The subject may be a mammal, for example, a human, a rat, a rabbit, a sheep, a pig, a cow, a cat, a dog, a monkey, etc., preferably a human.
本发明的药物组合物，可以口服或非口服施用。施用量因症状程度、患者年龄、性别、体重、敏感性差异、施用方法、施用时期、施用间隔、药物制剂的性质、有效成分的种类等而异，无特殊限制，但通常成人(体重60Kg)每日1μg～30000mg、优选10μg～3000mg、更优选100μg～2000mg、更优选1mg～1000mg、更优选5～900mg、更优选10mg～500mg、更优选100mg～300mg(以人参皂苷Rk3和人参皂苷Rh4总量计)，上述施用量通常可每日分1～3次施用。The pharmaceutical composition of the present invention can be administered orally or parenterally. The dosage varies depending on the degree of symptoms, patient age, sex, weight, differences in sensitivity, application method, application period, application interval, nature of the pharmaceutical preparation, types of active ingredients, etc., and there is no particular limitation, but usually adults (weight 60Kg) Daily 1μg～30000mg, preferably 10μg～3000mg, more preferably 100μg～2000mg, more preferably 1mg～1000mg, more preferably 5～900mg, more preferably 10mg～500mg, more preferably 100mg～300mg (total of ginsenoside Rk3 and ginsenoside Rh4 Meter), the above application amount can usually be applied 1 to 3 times a day.
实施例Example
下面通过具体的实施例对本发明作进一步说明,以下实施方式只以举例的方式描述本发明，但本发明的保护范围并不局限于此。The present invention will be further illustrated by specific examples below. The following embodiments only describe the present invention by way of example, but the protection scope of the present invention is not limited thereto.
实施例1 具有防治酒精性脂肪肝的人参皂苷Rk3/Rh4组合物的片剂制备Example 1 Preparation of tablets with ginsenoside Rk3/Rh4 composition for preventing and treating alcoholic fatty liver
取纯度≥98％的人参皂苷Rk3 100g，纯度≥98％的人参皂苷Rh4 200g，以及制备片剂常用的药用辅料500g，混合均匀后压片，干燥，包装，得到本发明的片剂2000片，每片含有效成分150mg。Take 100g of ginsenoside Rk3 with a purity of ≥98%, 200g of ginsenoside Rh4 with a purity of ≥98%, and 500g of medicinal excipients commonly used in the preparation of tablets, mix them uniformly, press, dry, and package to obtain 2000 tablets of the present invention , Each tablet contains 150mg of active ingredient.
实施例2 具有防治酒精性脂肪肝的人参皂苷Rk3/Rh4组合物的胶囊剂制备Example 2 Preparation of capsules with ginsenoside Rk3/Rh4 composition for preventing and treating alcoholic fatty liver
取纯度≥98％的人参皂苷Rk3 100g，纯度≥98％的人参皂苷Rh4 100g，以及常用的药用辅料600g混合均匀，再装入目前市场上销售的胶囊，得到本发明的胶囊剂2000粒，每粒胶囊中有效成分含量为100mg。Take 100g of ginsenoside Rk3 with a purity of ≥98%, 100g of ginsenoside Rh4 with a purity of ≥98%, and 600g of commonly used pharmaceutical excipients, mix them evenly, and then fill them into capsules currently on the market to obtain 2000 capsules of the present invention. The effective ingredient content in each capsule is 100mg.
实施例3 具有防治酒精性脂肪肝的人参皂苷Rk3/Rh4组合物口服液制备Example 3 Preparation of ginsenoside Rk3/Rh4 composition oral liquid capable of preventing and treating alcoholic fatty liver
取纯度≥98％的人参皂苷Rk3 40g，纯度≥98％的人参皂苷Rh4 80g，混合均匀，再采用常规方法加入适量调味剂结晶果糖、水苏糖、柠檬酸、果胶、稀释剂水，加热溶解，均质灌装，灭菌，制成本发明的口服液1000瓶，每瓶口服液中含有效成分120mg。Take 40g of ginsenoside Rk3 with a purity of ≥98% and 80g of ginsenoside Rh4 with a purity of ≥98%, and mix them evenly, then add appropriate amount of flavoring crystalline fructose, stachyose, citric acid, pectin, diluent water, and heat Dissolve, uniformly fill, and sterilize to prepare 1,000 bottles of the oral liquid of the present invention, and each bottle of the oral liquid contains 120 mg of active ingredients.
实施例4 人参皂苷Rk3/Rh4组合物防治酒精性脂肪肝的的功效验证试验Example 4 Efficacy verification test of ginsenoside Rk3/Rh4 composition in preventing and treating alcoholic fatty liver
成年雄性小鼠(18－22克)100只，随机分为正常对照组；模型组；给药剂量组：实施例2中本发明组合物1组(Rk3 15mg/kg，Rh4 15mg)；本发明组合物2组(Rk3 7.5mg/kg，Rh4 22.5mg)，本发明组合物3组(Rk3 22.5mg/kg，Rh4 7.5mg)、人参皂苷Rk3(30mg/kg)组、人参皂苷Rh4(30mg/kg)组、多烯磷脂酰胆碱(30mg/kg)阳性药对照组、人参皂苷Rg1(30mg/kg) 组、人参皂苷CK/Rh1(CK 15mg/kg，Rh1 15mg/kg)组共10组，除正常对照组外，其余各组小鼠每日灌胃给予50％乙醇1次，乙醇剂量为12mL/kg BW，连续4周。正常对照组小鼠灌胃给予等体积的纯净水。受试药物用纯净水配制成混悬液，每日灌胃给予1次，正常对照组与模型对照组小鼠灌胃给予等量的纯净水，灌胃体积为10ml/kg/d。试验过程中小鼠自由摄食饮水，每周称重两次，按体重调整受试样品量。模型对照组和各样品组于实验结束时一次灌胃给予50％的乙醇，小鼠灌胃量12ml/kg BW，阴性对照组给予纯净水，禁食16h。动物腹腔注射60mg/kg BW的戊巴比妥钠溶液麻醉后腹主动脉采血，并取肝组织，进行各项指标的检测：血清ALT、AST和LDL、血清TC、血清TG和LDL含量、肝组织TG含量及病理组织学检查。试验结果以均数加减标准差
Figure PCTCN2021088142-appb-000001
表示，统计学处理方法采用单因素方差分析；计算两药相互作用指数CI＝AB/(A×B)，T为加药组测定指标值，C为模型组测定指标值，AB为两药联用组的T/C值，A、B为药物单独作用组的T/C值。CI＜1时，两药有协同作用，CI≤0.7时，协同作用非常显著(参照David H.Kern,Carol R.Morgan,and Susanne U.Hildebrand-Zanki.In vitro and in vivo interaction between cisplatin and topotecan in ovarian carcinoma systems[J].Cancer Research,1988,48.以及李兴起.力达霉素对神经胶质瘤抑制作用及联合替莫唑胺的协同作用研究[D].中国协和医科大学,2009.)。 100 adult male mice (18-22 grams) were randomly divided into normal control group; model group; dosage group: group 1 of the composition of the present invention in Example 2 (Rk3 15mg/kg, Rh4 15mg); the present invention Composition 2 groups (Rk3 7.5mg/kg, Rh4 22.5mg), composition 3 groups of the present invention (Rk3 22.5mg/kg, Rh4 7.5mg), ginsenoside Rk3 (30mg/kg) group, ginsenoside Rh4 (30mg/ kg) group, polyene phosphatidylcholine (30mg/kg) positive drug control group, ginsenoside Rg1 (30mg/kg) group, ginsenoside CK/Rh1 (CK 15mg/kg, Rh1 15mg/kg) group, 10 groups Except for the normal control group, mice in the other groups were given 50% ethanol once a day by intragastric administration at a dose of 12 mL/kg BW for 4 consecutive weeks. The mice in the normal control group were given an equal volume of purified water by gavage. The test drug was formulated into a suspension with purified water, and was given by intragastric administration once a day. The mice in the normal control group and the model control group were given the same amount of purified water by intragastric administration, and the intragastric volume was 10ml/kg/d. During the experiment, the mice were free to eat and drink, weighed twice a week, and adjusted the amount of test samples according to their body weight. The model control group and each sample group were given 50% ethanol once at the end of the experiment. The mice were given 12ml/kg BW by gavage. The negative control group was given purified water and fasted for 16h. Animals were intraperitoneally injected with 60 mg/kg BW pentobarbital sodium solution anesthetized from the abdominal aorta, and blood was collected from the abdominal aorta, and liver tissues were taken for detection of various indicators: serum ALT, AST and LDL, serum TC, serum TG and LDL content, liver Tissue TG content and histopathological examination. The test result is the mean plus or minus the standard deviation
Figure PCTCN2021088142-appb-000001
Indicates that the statistical processing method adopts single-factor analysis of variance; calculate the two-drug interaction index CI=AB/(A×B), T is the measured index value of the drug addition group, C is the measured index value of the model group, and AB is the two-drug combination Use the T/C value of the group, A and B are the T/C values of the drug alone group. When CI <1, the two drugs have a synergistic effect, when CI ≤ 0.7, the synergy is very significant (see David H. Kern, Carol R. Morgan, and Susanne U. Hildebrand-Zanki. In vitro and in vivo interaction between cisplatin and topotecan in ovarian carcinoma systems[J].Cancer Research,1988,48. and Li Xingqi.Study on the inhibitory effect of lidamycin on glioma and the synergistic effect of combined temozolomide[D]. Peking Union Medical College, 2009.).
实验结果如下：The experimental results are as follows:
表1本发明组合物对小鼠体重、肝重、肝脏系数的影响
Figure PCTCN2021088142-appb-000002
Table 1 Effect of the composition of the present invention on mouse body weight, liver weight and liver coefficient
Figure PCTCN2021088142-appb-000002
注：**P≤0.01，*P≤0.05vs空白对照； △△P≤0.01， △P≤0.05vs模型组 Note: **P≤0.01, *P≤0.05vs blank control; △△ P≤0.01, △ P≤0.05vs model group
Figure PCTCN2021088142-appb-000003
Figure PCTCN2021088142-appb-000003
各组的体重差异无统计学意义(P≥0.05)；与空白组对比，模型组的肝质 量及肝脏系数明显增大(P≤0.01)，与模型组对比，各给药组的肝脏系数均下降显著(P≤0.05)，其中Rk3/Rh4组的肝脏系数下降极显著(P≤0.01)，本试验结果表明，各给药组均对酒精导致的小鼠肝脏肿大有明显的抑制作用，且相同剂量下本发明组合物1较各单一组分效果更好，具有协同增效作用(CI值＜1)，与多烯磷脂酰胆碱组、人参皂苷Rg1以及人参皂苷CK/Rh1相比本发明组合物1的肝脏系数下降效果更为显著。The weight difference of each group was not statistically significant (P≥0.05); compared with the blank group, the liver mass and liver coefficient of the model group were significantly increased (P≤0.01), compared with the model group, the liver coefficients of each administration group were all Significant decrease (P≤0.05). Among them, the liver coefficient of Rk3/Rh4 group dropped extremely significantly (P≤0.01). The results of this test showed that each administration group had a significant inhibitory effect on alcohol-induced liver enlargement in mice. And under the same dosage, the composition 1 of the present invention has a better effect than each single component, and has a synergistic effect (CI value <1), compared with the polyene phosphatidylcholine group, ginsenoside Rg1 and ginsenoside CK/Rh1 The liver coefficient reduction effect of the composition 1 of the present invention is more significant.
表2本发明组合物对酒精性脂肪肝小鼠血清ALT、AST活性的影响
Figure PCTCN2021088142-appb-000004
Table 2 The effect of the composition of the present invention on the activity of serum ALT and AST in alcoholic fatty liver mice
Figure PCTCN2021088142-appb-000004
Figure PCTCN2021088142-appb-000005
Figure PCTCN2021088142-appb-000005
注：**P≤0.01，*P≤0.05vs空白对照； △△P≤0.01， △P≤0.05vs模型组 Note: **P≤0.01, *P≤0.05vs blank control; △△ P≤0.01, △ P≤0.05vs model group
ALT、AST是肝脏损伤的标志性指标，正常血清中ALT、AST含量非常少，只有肝损伤后，肝细胞内的ALT、AST受到细胞破裂而释放到血液中，乙醇经肝脏代谢过程中产生的活性氧会直接对肝细胞造成氧化损伤。从表2看出，各给药组动物血清ALT和AST活性与模型组均有显著性差异(P≤0.05)，其中本发明组合物Rk3/Rh4组动物血清ALT和AST活性与模型组差异极显著(P≤0.01)，非常明显的低于模型组，而与空白对照组没有显著性差异(P＞0.05)，本试验结果表明，本发明组合物可明显降低酒精性脂肪肝小鼠血清中异常升高的ALT、AST的活性，相同剂量下本发明组 合物1较各单一组分以及多烯磷脂酰胆碱组、人参皂苷Rg1、人参皂苷CK/Rh1在相同剂量下对ALT、AST的活性降低效果更强，经过计算，人参皂苷Rk3与人参皂苷Rh4组合物1联用指数CI＜1，而组合物2、组合物3的联用指数CI＞1，说明本发明组合物1有良好的协同增效作用，对饮酒导致的肝细胞损伤具有良好的保护作用。ALT and AST are the hallmark indicators of liver damage. The content of ALT and AST in normal serum is very low. Only after liver damage, ALT and AST in liver cells are released into the blood by cell rupture, and ethanol is produced during liver metabolism. Reactive oxygen can directly cause oxidative damage to liver cells. It can be seen from Table 2 that the serum ALT and AST activities of animals in each administration group are significantly different from those of the model group (P≤0.05). Among them, the Rk3/Rh4 group of the composition of the present invention has extremely different serum ALT and AST activities from the model group. Significantly (P≤0.01), very significantly lower than the model group, but not significantly different from the blank control group (P>0.05). The results of this test show that the composition of the present invention can significantly reduce serum levels of alcoholic fatty liver mice Abnormally elevated ALT and AST activities, the composition 1 of the present invention at the same dose compared to each single component and polyene phosphatidylcholine group, ginsenoside Rg1, ginsenoside CK/Rh1 in the same dose of ALT, AST The activity reduction effect is stronger. After calculation, the combination index CI of ginsenoside Rk3 and ginsenoside Rh4 composition 1 is <1, and the combination index of composition 2 and composition 3 is CI> 1, indicating that the composition 1 of the present invention is good The synergistic effect of serotonin has a good protective effect on liver cell damage caused by drinking.
表3本发明组合物对酒精性脂肪肝小鼠血清TC、TG、LDL含量和肝脏组织TG含量的影响
Figure PCTCN2021088142-appb-000006
Table 3 The influence of the composition of the present invention on the serum TC, TG, LDL content and liver tissue TG content of alcoholic fatty liver mice
Figure PCTCN2021088142-appb-000006
Figure PCTCN2021088142-appb-000007
Figure PCTCN2021088142-appb-000007
注：**P≤0.01，*P≤0.05vs空白对照； △△P≤0.01， △P≤0.05vs模型组 Note: **P≤0.01, *P≤0.05vs blank control; △△ P≤0.01, △ P≤0.05vs model group
长期摄取酒精会使小鼠血清TC、TG含量异常升高，肝脏堆积胆固醇与三酸甘油酯，如表3所示，各给药组动物血清TC、TG、LDL含量及肝组织TG含量与模型组均有显著性差异(P≤0.05)，其中本发明组合物组动物血清TC、TG、LDL含量及肝组织TG含量与模型组差异极显著(P≤0.01)，经过计算，组合物1联用指数CI＜1，而组合物2、组合物3的联用指数CI＞1，说明本发明组合物1有良好的协同增效作用，而在本发明组合物比例范围之外，人参皂苷Rk3与人参皂苷Rh4没有协同增效作用，酒精对肝细胞的损伤是通过自由基介导的脂质过氧化作用实现的，本发明组合物1可以有效降低小鼠血清中的TC、TG含量，改善LDL水平，抑制内源性TC、TG的合成，改善脂质在体内的分布及在内脏器官的沉积。Long-term ingestion of alcohol will abnormally increase the serum TC and TG content in mice, and accumulate cholesterol and triglycerides in the liver. There are significant differences between the two groups (P≤0.05). Among them, the animal serum TC, TG, LDL content and liver tissue TG content of the composition group of the present invention are extremely different from the model group (P≤0.01). After calculation, the composition 1 With index CI<1, and the combination index CI of composition 2 and composition 3>1, it shows that the composition 1 of the present invention has a good synergistic effect, and outside the proportion of the composition of the present invention, ginsenoside Rk3 There is no synergistic effect with ginsenoside Rh4. The damage of alcohol to liver cells is achieved through free radical-mediated lipid peroxidation. The composition 1 of the present invention can effectively reduce the content of TC and TG in mouse serum and improve The level of LDL inhibits the synthesis of endogenous TC and TG, and improves the distribution of lipids in the body and the deposition of internal organs.
从小鼠肝脏病理切片可以明显看出来，对照组小鼠肝脏大小正常，呈红色、质软、边缘锐利；模型组小鼠肝脏明显增大，呈灰白色、有颗粒感、切 面带有油腻感；各治疗小组肝脏形态相较于模型组均有改善，但Rk3/Rh4组合物与R其他治疗小组相比肝脏形态更接近对照组。肝脏HE染色结果(图3，1-10)显示，对照组肝细胞大小形态正常，排列整齐，中央可见大而圆的核，细胞质着色均匀。肝细胞内无脂滴沉积，形态结构均正常，无脂肪变性及炎症改变；模型组肝细胞肿胀，界限不清。胞内可见大片空泡样脂滴,大小不一，发生脂肪变性的细胞呈灶状分布肝组织几乎被脂肪空泡取代，肝细胞气球样变性，汇管区内有炎性细胞浸润；人生皂苷Rk3、Rh4单独作用组、组合物2组、组合物3组、阳性药对照组、人参皂苷Rg1组以及人参皂苷CK/Rh1组肝组织内脂肪空泡散状分布，脂质堆积(油滴)变小，未见明显的炎症炎性细胞,。食喂本发明Rk3/Rh4组合物1的小鼠其肝组织内肝细胞内几乎无脂滴沉积，形态结构均接近正常，无脂肪变性，以上的效果以中剂量效果较佳，显示本发明所公开的Rk3/Rh4组合物能较好地改善酒精导致的小鼠肝脏脂肪变性程度。From the pathological section of the mouse liver, it can be clearly seen that the liver of the control group is normal in size, with red color, soft texture, and sharp edges; the liver of the model group is obviously enlarged, gray-white, grainy, and the cut surface is greasy; each The liver morphology of the treatment group was improved compared with the model group, but the liver morphology of the Rk3/Rh4 composition was closer to that of the control group than the other treatment groups of R. The results of HE staining of the liver (Figure 3, 1-10) showed that the liver cells in the control group were normal in size and morphology, neatly arranged, with a large and round nucleus visible in the center, and the cytoplasm was evenly colored. No lipid droplets were deposited in the liver cells, and the morphology and structure were normal, without steatosis and inflammation; the liver cells in the model group were swollen and the boundaries were unclear. There are large vacuole-like lipid droplets in the cell, with different sizes. The fatty degeneration cells are distributed in a focal shape. The liver tissue is almost replaced by fatty vacuoles. The hepatocytes are balloon-like degeneration, and there are inflammatory cell infiltration in the portal area; human saponin Rk3 , Rh4 single action group, composition 2 group, composition 3 group, positive drug control group, ginsenoside Rg1 group, and ginsenoside CK/Rh1 group. Fatty vacuoles in the liver tissue are distributed in scattered form, and lipid accumulation (oil droplets) changes Small, no obvious inflammatory inflammatory cells. The mice fed with the Rk3/Rh4 composition 1 of the present invention had almost no lipid droplet deposition in the liver cells in the liver tissue, the morphology and structure were close to normal, and there was no steatosis. The disclosed Rk3/Rh4 composition can better improve the degree of liver steatosis in mice caused by alcohol.
实施例5 人参皂苷Rk3/Rh4组合物防治酒精性脂肪肝的的机理探究Example 5 Research on the mechanism of ginsenoside Rk3/Rh4 composition in preventing and treating alcoholic fatty liver
测定实施例4中正常对照组；模型组；Rk3组，Rh4组，Rk3/Rh4组中的肝脏超氧化物歧化酶(SOD)、肝脏还原性谷胱甘肽(GSH)、和肝脏中丙二醛(MDA)水平；以及通过分子芯片筛选测定肝细胞中相关基因表达水平，试验结果以均数加减标准差
Figure PCTCN2021088142-appb-000008
表示，统计学处理方法采用单因素方差分析； The normal control group in Example 4; model group; Rk3 group, Rh4 group, Rk3/Rh4 group in liver superoxide dismutase (SOD), liver reduced glutathione (GSH), and liver malondioxide Aldehyde (MDA) level; and molecular chip screening to determine the expression level of related genes in hepatocytes, and the test results are based on the mean plus or minus the standard deviation
Figure PCTCN2021088142-appb-000008
Indicates that the statistical processing method uses one-way analysis of variance;
实验结果如下：The experimental results are as follows:
表4本发明组合物对酒精性脂肪肝小鼠肝脏SOD、GSH及MDA的影响
Figure PCTCN2021088142-appb-000009
Table 4 The effect of the composition of the present invention on the liver SOD, GSH and MDA of alcoholic fatty liver mice
Figure PCTCN2021088142-appb-000009
Figure PCTCN2021088142-appb-000010
Figure PCTCN2021088142-appb-000010
注：**P≤0.01，*P≤0.05vs空白对照； △△P≤0.01， △P≤0.05vs模型组 Note: **P≤0.01, *P≤0.05vs blank control; △△ P≤0.01, △ P≤0.05vs model group
酒精性脂肪肝的肝脏细胞极易受到氧化应激，模型组小鼠机体抗氧化能力减弱，表现为酒精代谢有害产物MDA水平升高，肝组织SOD活性下降，还原性谷胱甘肽GSH含量降低，与模型组比较，Rk3组，Rh4组，Rk3/Rh4组肝组织SOD活性、GSH含量极显著升高(P≤0.01)，MDA含量极显著降低(P≤0.01)，说明Rk3、Rh4具有抗氧化活性，可以有效改善饮酒导致的肝脏氧化损伤。The liver cells of alcoholic fatty liver are very susceptible to oxidative stress. The model group mice have weakened antioxidant capacity, which is manifested by increased levels of MDA, a harmful product of alcohol metabolism, decreased liver tissue SOD activity, and decreased levels of reduced glutathione GSH. Compared with the model group, the SOD activity and GSH content of liver tissue in the Rk3 group, Rh4 group, Rk3/Rh4 group were extremely significantly increased (P≤0.01), and the MDA content was extremely significantly reduced (P≤0.01), indicating that Rk3 and Rh4 have anti- Oxidative activity can effectively improve liver oxidative damage caused by drinking.
表5本发明组合物对酒精性脂肪肝小鼠重要代谢途径相关基因相对表达量表达统计表Table 5 The composition of the present invention has a statistical table showing the relative expression of genes related to important metabolic pathways in alcoholic fatty liver mice
Figure PCTCN2021088142-appb-000011
Figure PCTCN2021088142-appb-000011
注：表中，↓相应基因在肝细胞mRNA表达中下调倍数，↑表示相应基因在肝细胞mRNA表达中上调倍数。Note: In the table, ↓ the corresponding gene is down-regulated fold in liver cell mRNA expression, ↑ indicates the corresponding gene is up-regulated fold in liver cell mRNA expression.
脂肪酸合成酶(Fasn)，在脂肪酸合成中起着至关重要的作用，负责乙酰辅酶A和丙二酰辅酶A从头合成一种长链软脂酸(棕榈酸酯)的所有催化步骤，该基因的表达直接影响着脂肪酸合成酶多少，对控制动物脂肪沉积起到重要作用。脂蛋白脂肪酶(LpL)，是脂肪代谢的关键酶之一，作用于脂蛋白，主要分解脂蛋白中的三酰甘油，是血浆中清除三酰甘油的限速酶，并促使脂蛋白转移胆固醇、磷脂和载脂蛋白；LpL还具有增加乳糜微粒结合到LP受体上的能力，促进乳糜微粒摄取。固醇辅酶A去饱和酶1(Scd1)，是单不饱和脂肪酸生物合成的限速酶，在脂肪酸代谢中起中心调节作用，也是瘦素(Leptin)作用的目的基因之一；瘦素与糖尿病、肥胖、脂肪肝有着密切关系，是近年来代谢性疾病的热点领域之一。Fatty acid synthase (Fasn), which plays a vital role in fatty acid synthesis, is responsible for all the catalytic steps for the de novo synthesis of acetyl-CoA and malonyl-CoA to a long-chain palmitate (palmitate). This gene The expression directly affects the amount of fatty acid synthase and plays an important role in controlling animal fat deposition. Lipoprotein lipase (LpL) is one of the key enzymes of fat metabolism. It acts on lipoproteins and mainly decomposes triacylglycerols in lipoproteins. It is the rate-limiting enzyme for removing triacylglycerols in plasma and promotes the transfer of cholesterol from lipoproteins. , Phospholipids and apolipoprotein; LpL also has the ability to increase the ability of chylomicrons to bind to LP receptors, and promote the uptake of chylomicrons. Sterol-CoA desaturase 1 (Scd1) is the rate-limiting enzyme of monounsaturated fatty acid biosynthesis. It plays a central regulatory role in fatty acid metabolism and is also one of the target genes of leptin; leptin and diabetes , Obesity and fatty liver are closely related and are one of the hotspots of metabolic diseases in recent years.
本实验条件下模型组能够使Fasn合成增加，LpL合成减少，是导致肝细胞脂肪过量沉积的罪魁祸首，造成脂肪肝发生的首要因素；而给药各组均可以减少Fasn的合成，给药组Rh4以及本组合物Rk3/Rh4能显著增加LpL合成，减少肝组织脂肪酸合成，加强脂肪分解，对缓解肝脏脂肪变有积极意义。另外，模型组上调Scd1基因表达，可能是造成酒精性脂肪变的最主要分子机理之一，各给药组均却能够显著下调Scd1基因表达。Under the experimental conditions, the model group can increase the synthesis of Fasn and decrease the synthesis of LpL, which is the culprit leading to excessive fat deposition in liver cells and the primary factor in the occurrence of fatty liver; and each group of administration can reduce the synthesis of Fasn, the administration group Rh4 and the composition Rk3/Rh4 can significantly increase LpL synthesis, reduce fatty acid synthesis in liver tissue, strengthen lipolysis, and have a positive effect on alleviating liver fatness. In addition, the up-regulation of Scd1 gene expression in the model group may be one of the most important molecular mechanisms that cause alcoholic steatosis, but each administration group can significantly down-regulate the expression of Scd1 gene.
从以上的药效学实验可知，本发明组合物具有预防和治疗酒精性脂肪肝的作用，并且长期服用没有副作用。From the above pharmacodynamic experiments, it can be known that the composition of the present invention has the effect of preventing and treating alcoholic fatty liver, and it has no side effects after long-term use.
最后需要说明的是，上述描述仅仅为本发明的优选实施例，本领域的普通技术人员在本发明的启示下，在不违背本发明宗旨及权利要求的前提下，可以做出多种类似的表述，这样的变换均落入本发明的保护范围之内。Finally, it should be noted that the above description is only a preferred embodiment of the present invention. Under the enlightenment of the present invention, those skilled in the art can make a variety of similar methods without violating the purpose and claims of the present invention. It is stated that such transformations fall within the protection scope of the present invention.
工业实用性Industrial applicability
本发明提供了一种包含人参皂苷Rk3和Rh4的、协同增效的、具有预防和/或治疗酒精性脂肪肝作用的药物组合物。The present invention provides a pharmaceutical composition containing ginsenosides Rk3 and Rh4, which is synergistic and has the effect of preventing and/or treating alcoholic fatty liver.

Claims (10)

  1. 一种可用于预防和/或治疗酒精性脂肪肝的药物组合物，其包含作为活性成分的预防和/或治疗有效量的人参皂苷Rk3和人参皂苷Rh4、以及药学可接受的载体，A pharmaceutical composition that can be used for the prevention and/or treatment of alcoholic fatty liver, which comprises preventive and/or therapeutically effective amounts of ginsenoside Rk3 and ginsenoside Rh4 as active ingredients, and a pharmaceutically acceptable carrier,
    该药物组合物中，所述人参皂苷Rk3和人参皂苷Rh4的重量比为1:0.5～2、优选1:0.9～1.1。In the pharmaceutical composition, the weight ratio of ginsenoside Rk3 and ginsenoside Rh4 is 1:0.5-2, preferably 1:0.9-1.1.
  2. 根据权利要求1所述的药物组合物，所述药物组合物中，所述人参皂苷Rk3和人参皂苷Rh4的重量比为1：1。The pharmaceutical composition according to claim 1, wherein the weight ratio of the ginsenoside Rk3 and ginsenoside Rh4 in the pharmaceutical composition is 1:1.
  3. 根据权利要求1所述的药物组合物，以所述药物组合物中包含的总人参皂苷为100重量份计，所述药物组合物中包含的人参皂苷Rk3和人参皂苷Rh4合计为10重量份以上。The pharmaceutical composition according to claim 1, based on 100 parts by weight of total ginsenosides contained in the pharmaceutical composition, the total of ginsenoside Rk3 and ginsenoside Rh4 contained in the pharmaceutical composition is 10 parts by weight or more .
  4. 根据权利要求1所述的药物组合物，以所述药物组合物中包含的总人参皂苷为100重量份计，所述药物组合物中包含的人参皂苷Rk3和人参皂苷Rh4合计为100重量份。The pharmaceutical composition according to claim 1, based on 100 parts by weight of total ginsenosides contained in the pharmaceutical composition, the total of ginsenoside Rk3 and ginsenoside Rh4 contained in the pharmaceutical composition is 100 parts by weight.
  5. 根据权利要求1所述的用途，其中，所述药物组合物不包含除人参皂苷Rk3和人参皂苷Rh4之外的其它预防和/或治疗酒精性脂肪肝的有效成分。The use according to claim 1, wherein the pharmaceutical composition does not contain other effective ingredients for preventing and/or treating alcoholic fatty liver except ginsenoside Rk3 and ginsenoside Rh4.
  6. 人参皂苷Rk3和人参皂苷Rh4在制备用于预防和/或治疗酒精性脂肪肝的药物组合物中的用途，其中，该药物组合物中，所述人参皂苷Rk3和人参皂苷Rh4的重量比为1:0.5～2、优选1:0.9～1.1。Use of ginsenoside Rk3 and ginsenoside Rh4 in the preparation of a pharmaceutical composition for preventing and/or treating alcoholic fatty liver, wherein, in the pharmaceutical composition, the weight ratio of ginsenoside Rk3 and ginsenoside Rh4 is 1 : 0.5-2, preferably 1:0.9-1.1.
  7. 根据权利要求6所述的用途，其中，所述药物组合物中，所述人参皂苷Rk3和人参皂苷Rh4的重量比为1：1。The use according to claim 6, wherein, in the pharmaceutical composition, the weight ratio of ginsenoside Rk3 and ginsenoside Rh4 is 1:1.
  8. 根据权利要求6所述的用途，其中，以所述药物组合物中包含的总人参皂苷为100重量份计，所述药物组合物中包含的人参皂苷Rk3和人参皂苷Rh4合计为10重量份以上。The use according to claim 6, wherein, based on 100 parts by weight of the total ginsenosides contained in the pharmaceutical composition, the total of ginsenoside Rk3 and ginsenoside Rh4 contained in the pharmaceutical composition is more than 10 parts by weight .
  9. 根据权利要求6所述的用途，以所述药物组合物中包含的总人参皂苷为100重量份计，所述药物组合物中包含的人参皂苷Rk3和人参皂苷Rh4合计为100重量份。The use according to claim 6, based on 100 parts by weight of total ginsenosides contained in the pharmaceutical composition, the total of ginsenoside Rk3 and ginsenoside Rh4 contained in the pharmaceutical composition is 100 parts by weight.
  10. 根据权利要求6所述的用途，其中，所述药物组合物不包含除人参皂苷Rk3和人参皂苷Rh4之外的其它预防和/或治疗酒精性脂肪肝的有效成分。The use according to claim 6, wherein the pharmaceutical composition does not contain other effective ingredients for preventing and/or treating alcoholic fatty liver except ginsenoside Rk3 and ginsenoside Rh4.
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150019337A (en) * 2013-08-13 2015-02-25 주식회사 한국인삼공사 Pharmaceutical compositions for anti-inflammation or anti-oxidation containing ginsenoside rh4-enriched extraction
CN106109483A (en) * 2016-07-29 2016-11-16 陕西巨子生物技术有限公司 There is the glycol group/triol group rare ginsenoside compositions of anti-tumor activity
CN108685931A (en) * 2018-07-02 2018-10-23 南京类人生物材料有限公司 Panaxsaponin composition with hypoglycemic activity and its application
CN109394703A (en) * 2018-07-05 2019-03-01 西安巨子生物基因技术股份有限公司 3beta,6alpha,12beta-Trihydroxydammar-20's intravenous formulations and preparation method
CN111388490A (en) * 2020-04-17 2020-07-10 陕西巨子生物技术有限公司 Ginsenoside composition with function of preventing and treating alcoholic fatty liver

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103054906B (en) * 2012-11-30 2014-09-10 周斌 Propolis ethanol extract for alleviating hangover and preparation method thereof, and application of propolis ethanol extract in producing buccal tablets

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150019337A (en) * 2013-08-13 2015-02-25 주식회사 한국인삼공사 Pharmaceutical compositions for anti-inflammation or anti-oxidation containing ginsenoside rh4-enriched extraction
CN106109483A (en) * 2016-07-29 2016-11-16 陕西巨子生物技术有限公司 There is the glycol group/triol group rare ginsenoside compositions of anti-tumor activity
CN108685931A (en) * 2018-07-02 2018-10-23 南京类人生物材料有限公司 Panaxsaponin composition with hypoglycemic activity and its application
CN109394703A (en) * 2018-07-05 2019-03-01 西安巨子生物基因技术股份有限公司 3beta,6alpha,12beta-Trihydroxydammar-20's intravenous formulations and preparation method
CN111388490A (en) * 2020-04-17 2020-07-10 陕西巨子生物技术有限公司 Ginsenoside composition with function of preventing and treating alcoholic fatty liver

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BAEK SEUNG-HOON, SHIN BYONG-KYU, KIM NAM JAE, CHANG SUN-YOUNG, PARK JEONG HILL: "Protective effect of ginsenosides Rk3 and Rh4 on cisplatin-induced acute kidney injury in vitro and in vivo", JOURNAL OF GINSENG RESEARCH, vol. 41, no. 3, 1 July 2017 (2017-07-01), KR , pages 233 - 239, XP055857137, ISSN: 1226-8453, DOI: 10.1016/j.jgr.2016.03.008 *
QU LINLIN; ZHU YANYAN; LIU YANNAN; YANG HAIXIA; ZHU CHENHUI; MA PEI; DENG JIANJUN; FAN DAIDI: "Protective effects of ginsenoside Rk3 against chronic alcohol-induced liver injury in mice through inhibition of inflammation, oxidative stress, and apoptosis", FOOD AND CHEMICAL TOXICOLOGY, vol. 126, 1 January 1900 (1900-01-01), GB , pages 277 - 284, XP085632620, ISSN: 0278-6915, DOI: 10.1016/j.fct.2019.02.032 *

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