WO2009026794A1 - Composés de cyclohexénone issus de antrodia camphorata destinés à la protection du foie - Google Patents

Composés de cyclohexénone issus de antrodia camphorata destinés à la protection du foie Download PDF

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WO2009026794A1
WO2009026794A1 PCT/CN2008/070413 CN2008070413W WO2009026794A1 WO 2009026794 A1 WO2009026794 A1 WO 2009026794A1 CN 2008070413 W CN2008070413 W CN 2008070413W WO 2009026794 A1 WO2009026794 A1 WO 2009026794A1
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liver
compound
burdock
cyclohexenone compound
control group
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PCT/CN2008/070413
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Chinese (zh)
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Sheng-Yun Liu
Wu-Che Wen
Mao-Tien Kuo
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Golden Biotechnology Corporation
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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
    • A61K31/12—Ketones
    • A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
    • 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 a compound for protecting liver, in particular to a kind of
  • the liver is the most important and most important metabolic organ in the human body. It plays an important role in the metabolism of sugar, lipids, proteins, vitamins, hormones, bile and other substances. At the same time, the liver also has secretion, excretion, biotransformation and so on. Function; The liver is also an important barrier organ of the body. Its detoxification function has an important protective effect on the body. Therefore, when the liver function is damaged, it will cause metabolic disorders and affect other organs' functions, which is serious and life-threatening. The incidence of liver disease among Chinese people is very high. According to the statistics of the Department of Health of the Executive Yuan, chronic liver disease and cirrhosis are the sixth leading cause of death, and the number of deaths per year is more than 3,000, and the number is still rising. Therefore, it is imperative to develop substances that can effectively protect the liver and can be used to prevent or treat liver-related diseases.
  • Antrodia camphorata also known as Antrodia camphorata, burdock or red locust, etc.
  • Antrodia camphorata is a perennial fungus belonging to the genus Aphyllophorales and Polyporaceae. It is a unique species of fungi in Taiwan and only grows. On the inner wall of the hollow decayed heartwood of Cinnamoum kanehirai Hay in Taiwan. Due to the extremely rare distribution of burdock trees, artificially slashed, the number of wild burdocks that can grow in the middle of it is even rarer. And because its fruiting bodies grow quite slowly, the growth period is only between June and October, so the price is very expensive.
  • the fruiting body of Antrodia camphorata is perennial, sessile, with cork to wood. It has a strong aroma of eucalyptus, and its morphology varies, and it has a plate shape, a bell shape, a horseshoe shape or a tower shape. It is flat and bright red at the beginning of life, and then it radiates and rewinds around it, and grows to the periphery. The color also changes to reddish brown or yellowish brown, and there are many fine pores, and it is medicinal The most valuable part.
  • Niobium has the functions of detoxification, alleviating diarrhea, anti-inflammatory, treating liver-related diseases and anti-cancer.
  • Antrodia camphorata has many complex ingredients, such as triterpenoids and polysaccharides. It is known to have many complex components.
  • polysaccharides such as ⁇ -D-glucan
  • adenosine vitamins (such as vitamin B, nicotinic acid), proteins (including immunoglobulins), superoxide dismutase (SOD), trace amounts Elements (such as calcium, phosphorus, strontium), nucleic acids, sterols, and blood pressure stabilizing substances (such as antodia acid), which are considered to have anti-tumor, immune-enhancing, anti-allergic, anti-pathogenic, anti-hypertensive , lowering blood sugar, lowering cholesterol, protecting the liver and fighting fatigue.
  • Triterpenoids are a general term for a combination of thirty carbon elements into hexagonal or pentagonal natural compounds.
  • the bitter taste of Antrodia camphorata is mainly derived from triterpenoids.
  • Chemg et al. found that three extracts of triterpenoids based on ergostane were found in the extract of Antrodia camphorata fruit bodies: antcin A, antcin B and antcin C (Chemg, IH, and Chiang, HC 1995. Three new triterpenoids from Antrodia cinnamomea. J. Nat. Prod. 58:365-371). Chen et al. and E.
  • the present invention separates and purifies the compound of the formula (1) from the extract of Antrodia camphorata;
  • X oxygen (0) or sulfur (S)
  • Y oxygen or sulfur
  • R 2 is hydrogen, fluorenyl Or (CH 2 ) m -CH 3
  • the cyclohexenone compound of the formula (1) and the formula (2) in the present invention is isolated and purified from an aqueous extract of Antrodia camphorata or an organic solvent extract, and the organic solvent may include an alcohol (for example, decyl alcohol, ethanol or propanol), an ester ( For example, ethyl acetate), an alkane (e.g., hexane) or a halogenated alkane (e.g., chlorodecane, ethyl chloride), but not limited thereto, preferably an alcohol, and more preferably an ethanol.
  • an alcohol for example, decyl alcohol, ethanol or propanol
  • an ester For example, ethyl acetate
  • an alkane e.g., hexane
  • a halogenated alkane e.g., chlorodecane, ethyl chloride
  • the present invention is applied to liver protection and prevention and slowing of liver damage, and after feeding liver injury induced by carbon tetrachloride, feeding anthraquinone Cyclohexenone compounds help to slow the damage and fibrosis of rat liver tissue, and reduce alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in rat plasma.
  • the concentration in turn, achieves the efficacy of protecting the liver; this, as well as the concentration of catalase (CAT), in order to reduce the damage caused by free radicals to liver cells and the oxidative stress they bear, and improve the resistance of liver tissue Oxidation ability.
  • the compound of the formula (1) and/or the formula (2) can also be used as a component of a pharmaceutical composition for treating liver damage, thereby improving symptoms caused by liver damage in a mammal such as a human.
  • the aforementioned pharmaceutical composition may further comprise a pharmaceutically acceptable carrier in addition to an effective amount of a compound of the formula (1) and/or formula (2).
  • the carrier may be, but is not limited to, an excipient such as water, a filler such as sucrose or starch, a binder such as a cellulose derivative, a diluent, a disintegrant, an absorption enhancer or a sweetener. .
  • the pharmaceutical composition of the present invention can be produced according to a conventional pharmacy preparation method, and the active ingredient dose of the formula (1) and/or the formula (2) is mixed with one or more carriers to prepare a desired dosage form, and the dosage form is prepared.
  • carriers may include lozenges, powders, granules, capsules or other liquid preparations, but are not limited thereto.
  • Figure 1 is an example of the present invention, in the case of carbon tetrachloride-induced liver injury, anthraquinone cyclohexenone compound The effect on the weight of the rat.
  • control group ⁇ : negative control group (20% carbon tetrachloride); ⁇ : positive control group (20% carbon tetrachloride + milk thistle); country: treated with carbon tetrachloride and fed 300 Mg/kg experimental group of anthraquinone cyclohexenone compound;
  • A an experimental group treated with carbon tetrachloride and fed with 1000 mg/kg of anthraquinone cyclohexenone compound;
  • ⁇ treated with carbon tetrachloride and fed 3000 mg/ An experimental group of kg of anthraquinone cyclohexenone compound;
  • Fig. 2 is a graph showing the effect of the anthraquinone cyclohexenone compound on the food intake of rats in the case of liver injury induced by carbon tetrachloride in the examples of the present invention.
  • control group ⁇ : negative control group (20% carbon tetrachloride); ⁇ : positive control group (20% carbon tetrachloride + milk thistle); country: treated with carbon tetrachloride and fed 300
  • Experimental group of mg/kg anthraquinone cyclohexenone compound A: experimental group treated with carbon tetrachloride and fed with 1000 mg/kg of anthraquinone cyclohexenone compound; * : treated with carbon tetrachloride and fed 3000 mg/ An experimental group of kg of anthraquinone cyclohexenone compound;
  • FIG. 3 is a view showing the results of pathological changes of the liver surface of rats after the liver injury induced by carbon tetrachloride and the administration of anthraquinone cyclohexenone compound in the present invention.
  • Figure (A) is the control group (only corn oil is given);
  • Figure (B) is the negative control group (20% carbon tetrachloride is given);
  • Figure (C) is the positive control group (20% carbon tetrachloride and 200%) Mg/kg silybum);
  • Figure (D) is a low-dose anthrax ring cyclohexenone compound experimental group (20% carbon tetrachloride and 300 mg/kg anthraquinone cyclohexenone compound);
  • Figure (E) is medium Dosage of Antrodia camphora cyclohexenone compound experimental group (administer 20% carbon tetrachloride and 1000 mg/kg anthraquinone cyclohexenone compound);
  • FIG. 4 is a result of pathological changes of liver tissue damage in rats after liver injury induced by carbon tetrachloride according to an example of the present invention.
  • Figures (A) and (B) show the results of vigorous mitosis in swollen hepatocytes and hepatocytes after staining with hematoxylin-eosin staining (H&E stain) (magnification 200 X and 400 X)
  • Fig. (C) shows the vacuolar degeneration of liver tissue after staining with collagen staining (MT stain), and the liver fibrosis divides the liver parenchyma into many nodules, resulting in observation of cirrhosis ( The magnification is 100 X).
  • FIG. 5 is a view showing the pathological changes of liver fibrosis stained by collagen staining (MT stain) before and after liver injury induced by carbon tetrachloride in the Example of the present invention.
  • Figure (A) shows normal liver tissue
  • Figure (B) shows the degree of fibrotic lesions and collagen hyperplasia in liver tissue after treatment with carbon tetrachloride
  • Figure (C) shows the center of the liver after carbon tetrachloride treatment Vein and portal vein Incompletely septum
  • Figure (D) After treatment with carbon tetrachloride, the liver tissue forms a complete septum and intersects with each other, and divides the liver parenchyma into many nodules, but the septum is still thin
  • E) After treatment with carbon tetrachloride, the septum of the liver tissue becomes thicker and forms cirrhosis
  • Figure (F) shows the liver fibrosis and green collagen after treatment with carbon tetrachloride.
  • the mycelium, fruiting body or a mixture of the two is taken from Antwdia camphorata, and extracted by water or an organic solvent by a conventional extraction method to obtain an aqueous extract of Antrodia camphorata or an organic solvent extract.
  • the organic solvent may include an alcohol (for example, decyl alcohol, ethanol or propanol), an ester (such as ethyl acetate), an alkane (such as hexane) or a halogen (for example, chlorodecane, ethyl chloride), but Not limited to this.
  • alcohols for example, decyl alcohol, ethanol or propanol
  • an ester such as ethyl acetate
  • an alkane such as hexane
  • a halogen for example, chlorodecane, ethyl chloride
  • the extracted aqueous extract of Antrodia camphorata or the organic solvent extract can be further separated and purified by high-performance liquid chromatography, and then each fraction is subjected to biochemical tests related to liver protection. Finally, the composition of the liver-protecting liquid is analyzed, and the components that may produce liver-protecting effects are subjected to biochemical experiments such as the effect of slowing liver damage. Finally, it was found that the compound of the formula (1) / formula (2) of the present invention has an effect of slowing liver damage to protect the liver.
  • the anthraquinone cyclohexenone compound is administered, and the degree of liver damage is detected.
  • degree of liver fibrosis alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations, and glutathione (GSH), glutathione peroxidase (GSHPx)
  • liver enzymes such as catalase (CAT), superoxide dismutase (SOD), liver antioxidant enzymes, etc., to determine the liver-protecting ability of the anthraquinone cyclohexenone compound.
  • Triene-2-cyclohexenone compound has reduced Effects of liver injury and fibrosis caused by academic liver injury, and alanine aminotransferase
  • Inflammation of liver function such as (ALT) and aspartate aminotransferase (AST) is a concentration in the blood, and also promotes glutathione peroxidase (GSHPx) and catalase in the liver.
  • ALT aspartate aminotransferase
  • AST aspartate aminotransferase
  • the collected Antrodia camphorata extract was analyzed by high performance liquid chromatography using a column of RP18, and decyl alcohol (A) and 0.1% to 0.5% aqueous acetic acid solution.
  • (B) as the mobile phase (the ratio of the solution is: 0-10 minutes, B ratio is 95% to 20%; 10-20 minutes, B ratio is 20% to 10%; 20-35 minutes, The B ratio is 10% to 90%; 35 to 40 minutes, B ratio is 10% to 95%), and is eluted at a rate of 1 ml per minute, and analyzed by an ultraviolet-visible full-wavelength detector.
  • the extract is concentrated and concentrated for 25 minutes to 30 minutes to obtain a pale yellow powdery solid product, which is 4-hydroxy-2,3-dimethoxy-6-mercapto-5 (3,7,11 - Tridecyl-2,6,10-dodecatriene)-2-cyclohexenone.
  • the molecular formula is C 24 H 38 0 4
  • the molecular weight is 390
  • the melting point (mp ) is 48 ° C to 52 ° C.
  • the nuclear magnetic resonance (NMR) analysis values are as follows: 1.51, 1.67, 1.71, 1.75, 1.94, 2.03, 2.07, 111, 2.25, 3.68, 4.05, 5.07 and 5.14.
  • liver injury Because the main factors causing liver disease are viral, alcoholic and chemical, and the pathological phenomenon in the animal model of the mouse is consistent with the human body, it is chemical liver injury. Therefore, the evaluation method is for chemistry. Assessment of liver function in patients with liver injury; experimental model of chronic liver injury induced by carbon tetrachloride (CC1 4 ), and biochemical values of liver injury and liver tissue sections were examined to investigate the treatment of anthraquinone cyclohexenone compound The effect on chronic liver injury in rats. Among them, the toxicity of carbon tetrachloride can cause hepatocyte necrosis, and further develop into fibrosis of liver. If it continues to be administered, it will cause irreversible cirrhosis.
  • liver microsomal enzymes can be activated by liver microsomal enzymes to form trichloromethyl radical (CC1 3 - ), which is combined with protein to cause protein synthesis to be blocked, and cause lipid catabolism, causing triglyceride in liver cells.
  • Accumulation in addition to the peroxide formed by the trichlorodecane radical, can also lead to lipid peroxidation and damage to the liver cell membrane, resulting in enzyme exudation in the liver and cytopathic necrosis.
  • the liver injury model of carbon tetrachloride is a simulation of human liver cirrhosis, and thus can be applied to evaluate the therapeutic effect of a drug or a food component for treating liver diseases such as liver fibrosis.
  • the test steps are detailed below:
  • the experimental animals were five-week-old rats (Sprague-Dawley, SD) purchased from Lesco Biotech Co., Ltd., and the rats were indwelled in the animal breeding room for two weeks after purchase, and the body weight was similar (about 220 g). To 270 g) healthy adult rats were tested. Before the experiment, the rats were divided into six groups, 12 rats in each group. The body weight of the rats was recorded for each dose, which was used as the basis for the subsequent experimental doses. The grouping experiments shown in Table 1 were used as the control group and the negative control. Groups, positive control groups, and three groups of experimental groups of 300 mg/kg, 1000 mg/kg, and 3000 mg/kg, respectively.
  • silymarin can alleviate the inflammatory reaction in the liver and reduce the degree of liver damage. It can also reduce the toxicity of the liver by neutralizing the toxicity of toxic substances or competing with the toxic substances. The amount of absorption of the substance; and the anti-oxidant effect of the milk thistle can protect the liver cells from free radical damage. Because of its many different liver damage studies, including a large number of animal experiments and some clinical experiments, it has been shown to have a very good function of protecting the liver. Therefore, it is recognized as a therapeutic drug for liver diseases in modern medicine, and The animal model of liver injury was also used as a drug in the positive control group. Table 1. Experimental animal group and its feeding quality and dosage
  • control group 0 0 0
  • B (negative control group) 2 ml/kg BW 0 0 *, ,
  • Group A in Table 1 is a control group, which is a gastric tube perfusion with corn oil (purchased from Sigma chemical co.); Group B to F is filled with 20% of corn oil by gastric tube perfusion (v/v) ) Carbon tetrachloride (purchased from Japan's Shijiujiu), each rat was given a dose of 2 ml/kg according to its body weight, and gastric perfusion was given 2 days a week (Tuesday, Thursday afternoon).
  • group C was prepared by gastric tube perfusion with physiological saline to prepare 200 mg / kg dose of milk thistle;
  • group D to F was separately perfused with gastric tube by the preparation of Example 1 and prepared with physiological saline 300 mg /kg, 1000 mg/kg and 3000 mg/kg doses of Antrodia camphorata 4-hydroxy-2,3-didecyloxy-6-mercapto-5 (3,7,11-tridecyl-2,6 , 10-dodecatriene)-2-cyclohexenone compound, and the amount of food in the C to F group was 10 ml/kg of the above substance per body weight, and was given 5 days a week (am) Gastric tube perfusion was continued for 8 weeks.
  • the results of the measured rat body weight and rat feeding during the experiment are shown in Fig. 1 and Fig. 2.
  • Figure 1 is a control group and oral administration of 20% (v/v) carbon tetrachloride and administration of silymarin or burdock ring.
  • Figure 1 shows that there is no significant difference between the groups in the initial body weight, 20% by oral administration.
  • (v/v) After one week of carbon tetrachloride treatment, the body weight of the rats in the control group and the experimental group showed a downward trend compared with the control group without carbon tetrachloride treatment; and at the eighth week, The final body weight measured by the negative control group given the milk thistle or the burdock cyclohexenone compound was 366.9 g, which was significantly lower than the final body weight of the control group of 448.7 g, indicating that the treatment of carbon tetrachloride caused the rat body.
  • the lesion causes the rat to lose weight.
  • the positive control group of oral milk thistle showed a final body weight of 375.8 g, and the weight loss was less than that of the negative control group.
  • the body weight of the rat decreased due to the influence of carbon tetrachloride, but the effect of the milk thistle Rats did not lose weight; instead, they were fed different concentrations of Antrodia camphorata
  • the rats weighed were 386.6 g, 365.1 g and 355.0 g, respectively.
  • the mg/kg of Antrodia camphora cyclohexenone compound can effectively alleviate the weight loss caused by carbon tetrachloride.
  • Figure 2 is a control group and oral administration of 20% (v/v) carbon tetrachloride and administration of silymarin or burdock ring. It can be seen from Fig. 2 that the feed consumption of rats in each group increased at the first 1-5 weeks after treatment, but after the fifth week, the feed consumption of each group began to decrease and showed a downward trend, which was induced by carbon tetrachloride. Rat liver injury affects the food intake of experimental animals.
  • Rats were sacrificed at the end of the eighth week, and the liver, kidney and spleen were weighed by laparotomy, and the measured liver, kidney and spleen weights were compared to the total weight of the rats from which the organs were obtained. The ratio (%) of these organs to the total weight of the rats was converted to observe the effects of carbon tetrachloride treatment and administration of anthraquinone cyclohexenone on the weight of liver, kidney and spleen. In addition, each experimental group The data are expressed as mean and standard deviation (mean ⁇ SD). One-way analysis of variance (ANOVA) is used. When the variance analysis results show significant differences, the LSD method is further used.
  • ANOVA One-way analysis of variance
  • the D data is the mean value and the standard machine difference (Mean ⁇ SD ).
  • the percentage (%) of liver, kidney and spleen weight and total body weight measured by carbon tetrachloride-treated negative control group without silymarin or burdock hexacene ketene compound were significant ( /? ⁇ 0.05 ) Higher than the control group, this shows that carbon tetrachloride has caused liver, kidney and spleen lesions, and the weight of these organs has increased; the liver, kidney and spleen weight indicated by the positive control group of oral milk thistle It was higher than the control group due to the influence of carbon tetrachloride, but the weight of these organs was still lower than that measured by the negative control group due to the liver protection effect of the milk thistle itself.
  • liver tissue sections were taken. According to the anatomical location of the liver, the five hepatic lobules were cut into half of the liver tissue and frozen in a freezing rejection at -80 °C for subsequent detection of antioxidant enzyme activity. The remaining liver tissue was fixed in a 10% neutral formalin solution for one week. After roughing and paraffin embedding, tissue sections of 2 ⁇ thickness were prepared and stained with hematoxylin-eosin (Hematoxyline).
  • -eosin satin, H&E observed fat accumulation, inflammation, cell necrosis and fibrosis, and special staining of collagen fibers (Masson's trichromc, MT) stained reticular fibers and collagen fibers to assess the extent of liver fibrosis, in general Various histopathological changes of liver injury after the above pathological staining were observed under an optical microscope (Opticphot-2, Nikon, Tokyo, Japan).
  • liver fibrosis For semi-quantitative analysis of liver fibrosis, it can be based on Gabriele (Gabriele, B. 1997. Silymarin retards collagen accumulation in early and advanced biliary fibrosis secondary to complete bile duct obliteration in rats. Hepatology 26: 643-649.) and Wang ( Wang, GS, Eriksson, L. C, Xia, L., Olsson, J. and Stal, P. 1999. Dietary iron overload inhibits carbon tetrachloride-induced promotion in chemical hepatocarcinogenesis: effects on cell proliferation, apoptosis, and antioxidation. J Hepatol.
  • the method of liver fibrosis is divided into the following 0 to 4 grades: Grade 0 represents normal liver tissue without any liver fibrosis, Grade 1 represents collagen hyperplasia, but no septum is formed (radial fibrosis in the central vein or portal vein), and grade 2 represents an incomplete septum between the central vein and the portal vein (in this case) Level 3 does not meet each other. Level 3 represents the formation of a complete septum, which intersects each other and divides the liver parenchyma into many nodules. However, the septum is still thin and grade 4 represents the formation of a complete septum, and the septum becomes thicker, that is, complete cirrhosis. The results are shown in Figures 3 to 5 and Table 3.
  • Fig. 3 is a view showing the observation of liver surface pathological changes in the control group and the experimental group of the control group and the carbon tetrachloride-induced liver injury and the administration of the milk thistle or the burdock.
  • the liver surface of the control group was smooth and complete (please refer to Fig.
  • liver fibrosis is divided into 0 ⁇ 4 grades, 0 is normal liver tissue, no liver Fibrosis, 1 is the proliferation of collagen, but does not form a septum (radial fibrosis in the central vein or portal area), 2 between the central vein and the portal vein, forming an incomplete septum (this septum) There is no meeting with each other), 3 In order to form a complete septum, the middle meets and divides the liver parenchyma into many nodules, but the septum is still thin and 4 is a complete septum and the septum becomes thicker, forming a complete liver. hardening.
  • Figure 4 shows the pathological changes of liver tissue damage after liver injury induced by carbon tetrachloride. As can be observed from Figure 4, after treatment with carbon tetrachloride, The tissue sections of the liver cells are swollen and cause vigorous mitosis of the liver cells.
  • Fig. 4A and Fig. 4B The number of Kuffer's cells will also increase (Fig. 4A and Fig. 4B), and in severe cases, vacuolization will occur, and the fibrotic liver will divide the liver into many nodules to form cirrhosis (Fig. 4C).
  • Fig. 4C the chronic liver injury assessment method published by Jonker et al.
  • the chronic liver injury induced by carbon tetrachloride was the most obvious, mainly distributed in 3 moderate ( + + + ) and 4 significant ( + + + + + ) level, pathological scores up to 3.2.
  • the degree of chronic liver injury was mild, mainly distributed at 3 moderate (+ + + ) level, and the pathological score was 2.8;
  • the experimental group of carbon tetrachloride treated and fed with different concentrations of Antrodia camphora cyclohexanone compound can improve the degree of chronic liver injury, and the dose of Antrodia camphora cyclohexenone compound is 300 mg/kg and 1000 mg/kg.
  • the group effect was most significant, mainly distributed in 1 mild ( + ) grade and 2 mild ( + + ) grades.
  • the pathological scores were reduced to 1.8 and 2.0, respectively.
  • Figure 5 shows the pathological changes of liver tissue fibrosis after hepatic injury induced by carbon tetrachloride. It can be observed from Fig. 5 that there is no fibrosis in normal liver tissue (see Fig. 5A), and the liver at this time belongs to grade 0 in the degree of fibrosis published by Gabriele and Wang et al; After carbon treatment, liver tissue has varying degrees of fibrotic lesions.
  • the collagen that proliferates but does not form a septum belongs to grade 1 in the degree of fibrosis, when the central vein and portal area
  • the liver that forms the incomplete septum between the two is Grade 2 in the degree of fibrosis, if the intact septum is formed and the middle meets and divides the liver parenchyma into many nodules, but this septum
  • the liver which is still very thin (see Figure 5D), is grade 3 in the degree of fibrosis, and once the formed septum becomes thicker, it becomes cirrhosis (see Figure 5E), where the liver is fibrosis.
  • the positive control group treated with carbon tetrachloride and feeding silymarin had a milder degree of liver fibrosis with a pathological score of 2.8; treated with carbon tetrachloride and fed with different concentrations of Antrodia camphorata
  • the experimental group of the hexenone compound can effectively reduce the degree of liver fibrosis,
  • the effect of the group of 300 mg/kg of Antrodia camphora cyclohexenone compound was the most significant, mainly distributed in grades 1 to 2.
  • the pathological product decreased to 1.8.
  • ALT and AST are important enzymes in various organs of the human body such as liver, heart and muscle, and are used to participate in the synthesis of important amino acids in the body. Under normal circumstances, these enzymes maintain a stable low amount in serum, wherein the normal value of alanine aminotransferase is below 40 U/L, and the normal value of aspartate aminotransferase is below 50 U/L.
  • transaminase and aspartate aminotransferase are released, so that the concentration of these enzymes in the serum is increased, so the alanine
  • the activity of transaminase and aspartate aminotransferase is related to liver inflammation and hepatocyte damage, and can be used as one of the indicators for evaluating liver diseases.
  • Rats were sacrificed at the end of the eighth week, and liver biochemical function was examined by blood sampling in the abdominal aorta. Place 5 ml of blood into a blood cell serum separation tube and centrifuge at 775 xg for 15 minutes. Take 0.5 ml of serum to a serum biochemical analyzer (Express plus automatic clinical chemistry analyzer, Chiron diagnostics corporation, OH, USA) and use the detectable valley. Liver enzyme activity and cholesterol (Bayer diagnostics, Cat No. E33940), such as ALT; Bayer diagnostics, Cat No. E36941, glutamate aminotransferase (AST; Bayer diagnostics, Cat No. E37041) The biochemical values of these liver damages were measured by commercial reagents.
  • the D data were averaged by MS-Excels method and the standard machine difference (Mean ⁇ SD).
  • the liver is damaged by carbon tetrachloride, resulting in an increase in liver damage enzyme activity in plasma, and a slight increase in cholesterol content; a positive control group treated with carbon tetrachloride and fed with milk thistle compared to the control group
  • the glutamate transaminase (ALT), glutamate aminotransferase (AST) and cholesterol values were higher than the negative control group, indicating that the milk thistle has reduced carbon tetrachloride.
  • ALT 181.7 Disabled 78.4 U / L
  • AST 283.2 ⁇ 144.2 U/L
  • Antioxidant enzymes are glutathione (GSH), glutathione peroxidase (GSHPx), catalase (CAT), superoxide dismutase (superoxide dismutase). , SOD ), etc., through the antioxidant system, can build a strict line of defense in the organism, in order to prevent the oxidation of cells in organ tissues and reduce the oxidative stress when free radicals increase in the body. ).
  • GSH glutathione
  • GSHPx glutathione peroxidase
  • CAT catalase
  • superoxide dismutase superoxide dismutase
  • SOD superoxide dismutase
  • the liver in the above-mentioned frozen rejection at -80 ° C was taken out, and immersed in a phosphate buffered saline solution (pH 7.4 ) containing 0.16 mg/ml heparin to remove red blood cells for homogenization.
  • the tissue was disrupted and 10 ml of buffer (50 mM phosphate, pH 6-7, containing 1 mM EDTA) was added to each gram of tissue, and the homogenizer (Brinkmann Polytron homogenizer, setting 5, PT 10 probe) was used. After crushing for 1 minute, at 4.
  • GSH Glutathion activity ( mM/g protein );
  • GSHPx Glutathion peroxidase activity ( nmol/min/ml ) One unit is defined as 25 ° C per One minute is defined as the amount of enzyme that will cause the oxidation of 1 nmol of NADPH to NADP+ per minute at 25 °C;
  • D data is the mean and standard machine difference (mean ⁇ SD),
  • the concentration of superoxide dismutase (SOD) enzyme exhibited was significantly lower (/? ⁇ 0.05) than other groups, and the feeding doses were 1000 mg/kg and 3000 mg/kg.
  • the content of glutathione peroxidase (GSHPx) and catalase (CAT) was slightly higher than other groups, of which glutathione peroxidase (GSHPx) was used.
  • Hydrogenase (CAT) is a water that can decompose hydrogen peroxide (H 2 O 2 ) into cells that are not toxic to cells, so as to reduce the damage of peroxide to tissues.
  • anthraquinone cyclohexenone compound By increasing the concentration of glutathione peroxidase (GSHPx) and catalase (CAT) in the liver, the damage caused by free radicals to hepatocytes and the oxidative stress they undergo can be reduced, and the resistance of liver tissue can be enhanced. Oxidation ability.
  • GSHPx glutathione peroxidase
  • CAT catalase
  • the present invention is isolated from 4-hydroxy-2,3-dimethoxy-6-mercapto-5 (3,7,11-tridecyl-2,6,10-dode carbon) of Antrodia camphorata Triene)-2-cyclohexenone compound, which can effectively alleviate liver tissue damage and liver fibrosis caused by chemical liver injury, and can reduce alanine aminotransferase (ALT) and aspartate aminotransferase in blood (AST) and other indicators of liver function inflammation and damage, while also increasing the content of glutathione peroxidase (GSHPx) and catalase (CAT) in the liver to reduce the damage caused by free radicals to liver cells.
  • ALT alanine aminotransferase
  • AST aspartate aminotransferase in blood
  • GSHPx glutathione peroxidase
  • CAT catalase
  • the anthraquinone cyclohexenone compound is a naturally extracted substance, it is used for treating liver damage or protecting the liver, and does not cause discomfort or other side effects such as toxicity, complications, etc. It is resistant to free radicals such as hydrogen peroxide (3 ⁇ 40 2 ), so it can be prepared into health foods, foods and beverages, etc., to prevent liver damage and improve human health. In addition, it can also be used to protect the human body.
  • the ketene compound is prepared as a pharmaceutical composition for mitigating liver damage caused by a chemical substance, wherein the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier in addition to an effective amount of the anthraquinone cyclohexenone compound.
  • the carrier may be an excipient (such as water), a filler (such as sucrose or starch), a binder (such as a cellulose derivative), a diluent, a disintegrant, an absorption enhancer or a sweetener, but is not limited thereto. .
  • the pharmaceutical composition of the present invention can be produced according to a conventional pharmacy preparation method, and an active ingredient dose of an antrodia camphora cyclohexenone compound is mixed with one or more carriers to prepare a desired dosage form, and the dosage form may include a tablet. , powders, granules, capsules or other liquid preparations, But not limited to this. In order to prevent and treat liver damage in mammals such as humans and protect the liver.

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  • Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
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  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Engineering & Computer Science (AREA)
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  • Medicines Containing Plant Substances (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)

Abstract

Cette invention concerne des composés d'acyclohexénone issus de Antrodia camphorata, en particulier la 4-hydroxy-2,3-diméthoxy-6-méthyl-5[3,7,11-triméthyl-dodéca-2,6,10-triényl]-cyclohex-2-énone, qui peuvent protéger le tissu hépatique d'une manière efficace. Dans l'invention, les composés d'acyclohexénone issus de Antrodia camphorata destinés à la protection du foie ont pour effets d'atténuer la lésion et la fibrose du foie induites par une lésion chimique du foie, de diminuer la concentration des enzymes ALT et AST, d'augmenter la concentration des enzymes GSHPx et CAT, de diminuer la lésion des cellules hépatiques induite par les radicaux libres, d'améliorer l'effet antioxydatif du tissu hépatique.
PCT/CN2008/070413 2007-08-29 2008-03-05 Composés de cyclohexénone issus de antrodia camphorata destinés à la protection du foie WO2009026794A1 (fr)

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CN2007101477925A CN101376624B (zh) 2007-08-29 2007-08-29 用于护肝的牛樟芝环己烯酮化合物

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CN102232944B (zh) * 2010-05-06 2013-03-13 国鼎生物科技股份有限公司 用于抑制口腔癌肿瘤细胞生长的牛樟芝环己烯酮化合物
CN104273540A (zh) * 2014-10-20 2015-01-14 中山安荞生物科技有限公司 一种牛樟芝的组合物

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1352990A (zh) * 2000-11-13 2002-06-12 葡萄王企业股份有限公司 樟芝菌丝体生物活性物质其制备法及含其组成物
CN1379082A (zh) * 2001-08-10 2002-11-13 吴丽玉 樟芝的固体培养方法,所得固体培养物及其产品与用途
CN1666982A (zh) * 2004-03-10 2005-09-14 善笙生物科技股份有限公司 来自牛樟芝菌丝体的新混合物和化合物及其用途
US7342137B1 (en) * 2007-01-08 2008-03-11 Golden Biotechnology Corporation Cyclohexenone compounds from Antrodia camphorata and application thereof

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Publication number Priority date Publication date Assignee Title
CN1352990A (zh) * 2000-11-13 2002-06-12 葡萄王企业股份有限公司 樟芝菌丝体生物活性物质其制备法及含其组成物
CN1379082A (zh) * 2001-08-10 2002-11-13 吴丽玉 樟芝的固体培养方法,所得固体培养物及其产品与用途
CN1666982A (zh) * 2004-03-10 2005-09-14 善笙生物科技股份有限公司 来自牛樟芝菌丝体的新混合物和化合物及其用途
US7342137B1 (en) * 2007-01-08 2008-03-11 Golden Biotechnology Corporation Cyclohexenone compounds from Antrodia camphorata and application thereof

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