KR100707917B1 - A pharmaceutical composition for the treatment of hepatopathy comprising beta-glucan as an effective ingredient - Google Patents

Abstract

The present invention relates to a composition for treating liver disease comprising β-glucan as an active ingredient, and a health functional food for enhancing liver function. Specifically, the β-glucan has a β-1,3 / 1,6 bond and is produced from Aureobasidium pullulans SM2001, and includes a liver including diabetic liver disease and subacute liver injury. It can be usefully used for the treatment of a condition.

Liver disease, β-glucan

Description

Pharmaceutical composition for treating liver disease comprising beta glucan as an active ingredient {A PHARMACEUTICAL COMPOSITION FOR THE TREATMENT OF HEPATOPATHY COMPRISING BETA-GLUCAN AS AN EFFECTIVE INGREDIENT}

1 is a photograph showing the histological changes between STZ induced diabetic liver disease rats. (a) Sham, (b) medium control group, (c) silymarin 25 mg / kg dose group, (d) β-glucan 62.5 mg / kg dose group and (e) 125 mg / kg dose group. H & E staining, Scale bars = 200um. In the media control group, hepatic fibrosis was noted, which was characterized by prominent connective tissue formation around the portal vein of the liver.

2 is a photograph showing the histological changes of livers of STZ rats classified according to the severity of liver disease. HC (a, b), HG (c), LC (d), and LG (e). Compared to the treatment with β-glucan (HG, c), typical diabetic hepatic findings are shown in the controls not treated with β-glucan (HC, a and b). H & E staining, Scale bars = 200um.

Figure 3 is a photograph showing the histological analysis between CCl4 induced subacute liver disease rats. (1) Sham, (2) media control, (3) CAPT, (4) SILY and glucan (5) 31.25, (6) 62.5 and (7) 125 mg / kg-administered group.

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Seljelid, R. A Water-soluble aminated β-1,3-D-glucan derivative causes regression of solid tumors in mice. Biosci. Rep. 1986: 6: 845-851

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CCl4 an adequate model of human cirrhosis? Hepatology. 1983; 3: 112-120

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The present invention belongs to the pharmaceutical composition and functional food, and specifically relates to a pharmaceutical composition for treating liver disease comprising β-glucan as an active ingredient and health functional food for improving liver function.

The liver is one of the important organs responsible for the body's metabolic processes [Meyer and Kulkarni, 2001], glucose metabolism, lipid metabolism, nucleic acid synthesis, activation and storage of various vitamins, bile acids, bilirubin metabolism and excretion, detoxification, blood coagulation It is involved in almost every metabolism in the body, including the production of factors, immune activity, and blood volume control.

The liver has a great ability to prevent and regenerate, but it does not prevent the invasion of the virus due to a decrease in immunity, or when exposed to toxins, pollutants, and the like, it causes hepatitis, liver cirrhosis and liver cancer. For example, the liver is damaged by localization and fibrosis, and it is known that liver localization and fibrosis are closely related to the occurrence of cirrhosis, which causes death [Ulin et al., 2003]. The pathogenesis of fibrosis, one of the cirrhosis factors, is not yet clear, but one clear fact is that free radicals play an important role in pathological changes of the liver, especially alcoholic liver disease and toxic liver disease [Poli and Parolar, 1997]. These free radical groups are induced to form by pro-oxidant contaminants of drug use, radioactivity and the environment. The free radicals thus formed promote fat peroxidation, and have a fatal effect on cell membranes whose lipids are lipids, leading to many symptoms. A representative material for forming free radical groups in the body is the industrial solvent carbon tetrachloride (CCl4), which is a well known hepatotoxic substance [Abraham et al., 1999; Guven et al., 2003; Szymonik-Lesiuk et al., 2003]. CCl4 forms trichloromethyl radicals, which are free radical groups, in the body by reductive dehalogenation with cytochrome p450 (Recknagel et al., 1989).

Liver disease also occurs as a complication in diabetics. Diabetes is largely divided into insulin-dependent (type 1 diabetes) and insulin-independent (type 2 diabetes). Despite the development of anti-diabetic agents, diabetic liver disease occurs in many patients of all types.

Liver diseases with various conditions as described above are gradually increasing, and as long as the symptoms are not severe, there are few subjective symptoms, and thus, early detection is difficult, and once damaged, it is difficult to recover. The number is increasing. In Korea, chronic liver disease, including liver cancer and cirrhosis, is a disease with the highest prevalence of the 5th highest hospitalization frequency (Statistics Annual Report, 1994). According to the 2004 Statistical Yearbook of Statistics, the mortality rate of liver disease per 100,000 population is 19.1, which is the sixth highest after diabetes, and the development of effective treatments is urgently needed.

Many agents for the treatment of liver disease, including cirrhosis and diabetic hepatic disease, are known.

Captopril (CAPT) is an angiotensin converting enzyme inhibitor (ACE). ACE is an important enzyme in the formation of angiotensin II, causing arterial contraction and raising blood pressure. Therefore, although CAPT has been mainly used as a blood pressure lowering agent, recent studies have reported that it is effective in liver fibrosis induced by CCl4 [El-Khatib and Mansour, 2001; Tuncer et al., 2003].

Silymarin is a flavonoid found in thistle milk thistle. Silymarin is a potent antioxidant that protects hepatocytes from toxins [Wellington and Jarvis, 2001] and has been reported to be effective in diabetic liver disease and liver fibrosis induced by CCl4 [Bhattacharyya et al., 2003; Achliya et al., 2004; Shailajan et al., 2005].

In addition, drugs that inhibit hepatic fibrosis and cirrhosis are penicillamine, 16, 16-dimethyl prostaglandin E2, biphenyl dimethyl dicarboxylic acid, colchicine, glucocorticoid, malotilate, gamma interferon, pentoxifylline (pentoxifylline), pyridine-2, 4-dicarboxylic-diethylamide, pyridine-2, 4-dicarboxylic-di (2-methoxyethyl) amide, and the like (Boker et al., 1991). Tamayo et al., 1983; Ala-Kokko et al., 1989; Guzelian et al., 1984).

However, the drug has a problem in that it is not only effective or severe side effects when applied to the clinical, but also expensive.

Korean Patent Publication No. 92-10763 describes polysaccharides derived from Ganoderma lucidum mushroom (Garoderma lucidum IY009) KFCC 10709. The polysaccharides described in this document are one of protein polysaccharides, and contain β-glucose, alpha-glucose, galactose, alpha-mannose, and fructose, and as glycoproteins, lysine, alinine, hastidine, arginine, varin, Aspartic acid, threonine, isoleucine, serine, leucine, glutamic acid, tyrosine, proline, phenylalanine, methionine, but there is a problem causing liver toxicity.

On the other hand, glucan is a natural substance widely applied industrially in various fields such as cosmetics, immune activators and anticancer agents in developed countries, and its stability in human use has been proved. Glucan is a complex polysaccharide in the form of fibrin to which glucose is linked and classified into alpha-glucan and β-glucan. The former is generally present in starch of plants, the latter is known to be present in the cell walls of cereals, mushrooms and yeasts, and β-glucan is widely used in various industrial fields such as anticancer agents.

Grain-derived β-glucans such as yeast, mushrooms, barley and oats show differences in properties within β-glucans, depending on the linkage structure and physical properties of glucose. For example, β-glucan derived from mushrooms isolated from the fruiting bodies of shiitake mushrooms is water-soluble, activates the complement system that exhibits immune functions such as infection defense in vivo, dramatically improves immune function, and is known to exert antitumor effect. Czop, 1986; Estrada et al., 1997], β-glucans present in the cell walls of grains such as barley and oats have been reported to have an excellent effect on the lowering of blood cholesterol [Lia et al., 1995; Bell et al., 1999, the use as a therapeutic agent for liver disease has not been disclosed.

Korean Patent Publication No. 484319 and Kruger et al. And Luegmayr et al. Disclose the use of β-glucan, but do not imply or disclose the use of hepatopathy.

Accordingly, the present invention has been completed by recognizing the problems of the conventional liver disease treatment agent as described above, and is effective in treating liver disease that can replace the same, while searching for natural substances safe for the human body, having a branched structure. The present invention was completed by finding that water-soluble β-glucan having a specific structure is effective in treating liver disease.

An object of the present invention is to provide a therapeutic agent for a liver disease derived from a natural substance and a health functional food which can be effectively used for the treatment of liver disease.

The present invention for achieving the above object provides a pharmaceutical composition for treating liver disease comprising β-glucan or derivatives thereof as an active ingredient.

The present invention also the β-glucan comprises a glucose main chain connected by β-1,3 bond, wherein at least one of the glucose is β-1,3 / 1, connected by branched glucose and β-1,6 bond, 6-glucan is provided.

The present invention also provides a composition wherein the β-glucan is derived from Aureobasidium pullulans SM2001.

The present invention also provides a health food for improving liver function comprising β-glucan or a derivative thereof as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for treating liver disease, comprising β-glucan or a derivative thereof as an active ingredient.

Glucan is a complex polysaccharide found in the cell walls of yeast, oats or barley, and mushrooms used for medical purposes, such as, for example, Maitake mushrooms, as mentioned in the background, and is derived from various plants and fungi derived from the fungus. -Glucans can be included in the compositions of the present invention.

In particular, the β-glucan comprises a glucose main chain connected by β-1,3 bonds, and at least one of the glucose is a water-soluble β-1,3 / 1 structure of branched glucose and β-1,6 bonds , 6-glucan or derivatives thereof.

Specifically, β-1,3 / 1,6-glucan is β-1,3 and β-1 in which one molecule of glucose is β-1,6 bound per 5 molecules of glucose connected by β-1,3 bonds. Branched β-glucan with 6 bonds mixed together. In addition to the β-1,3 bond, the physiological activity is higher than that of other linear β-glucans due to the β-1,6 bond.

The β-1,3 / 1,6-glucan has an amorphous structure with low crystallinity and has molecular weight ranging from hundreds of thousands to millions. In addition, the lactic acid group is substituted with glucose branched in the main chain, so that the water-soluble acidic polysaccharide is easily dissolved in water.

Water-soluble β-glucan has improved results in terms of physiological activity such as increased immunomodulatory capacity and antitumor activity than insoluble (Darina Slamenovaa, D. et al, 2003; Seljelid, R. A, 1986; Svegliati-Baroni G et al., 1999; Williams, DL et al, 1992; Zhanga, M. et al, 2004). The term derivative used in the present invention is a term known in the art, and the hydroxyl group of branched glucose is substituted with, for example, carboxymethyl group, sulfuric acid group, amine group, sulfoethyl group, etc. It is to include.

In particular, β-1,3 / 1,6-glucan having the above characteristics has the structure of Formula 1 below.

(In Formula 1, n is 1 to about 4,000 and X is a lactic acid group, and other examples thereof include maleic acid or sulfoacetic acid.)

More specifically, the glucan is Aureobasidium Aureobasidium of the applicant's Korean Patent Registration No. 0443209 pullulans ) is a variant of Aureobasidium pullulans SM-2001 (KCCM 10307), and can be prepared according to the methods of preparing β-glucans known in the art or described in the patent specification.

In one embodiment of the present invention, Streptozotocin (STZ) -induced diabetic liver disease rat model and CCl4-induced subacute liver injury rat model, which is a representative animal model of liver disease, was prepared, and the rats were treated with aureobasidium pullulans. Administration of β-1,3 / 1,6-glucan from SM-2001 significantly reduced hepatic fibrosis and liver damage as compared to untreated controls, and blood geothyroid and gephyti levels, indicators of liver function A significant decrease in was confirmed.

As used herein, the term “liver disease” is meant to include acute or chronic liver disease, including but not limited to abnormalities in some or all of liver function, including but not limited to alcoholic liver disease, toxic liver disease, chronic And acute hepatitis, liver fibrosis and cirrhosis, inflammatory liver disease, diabetic liver disease.

The β-glucan or derivatives thereof of the present invention may be administered in the form of a pharmaceutically acceptable salt, or may be administered alone or in combination with other pharmaceutically active ingredients to enhance efficacy as a therapeutic agent for liver disease.

In addition, pharmaceutical compositions comprising β-glucan or derivatives thereof of the present invention are suitable carriers, diluents, preservatives, stabilizers, wetting agents, emulsifiers, solubilizers, sweeteners, colorants, osmotic pressure regulators, antioxidants commonly used in the manufacture of pharmaceutical compositions. It may further include excipients. Specifically, lactose, dextrose, sucrose, sorbitol, mannitol xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrroly Dough, water, methylhydroxy benzoate, propyl hydroxy benzoate, talc, magnesium, stearate, mineral oil, etc. are mentioned.

The method of administering the pharmaceutical composition for treating liver disease containing β-glucan of the present invention may be easily selected according to the formulation, and may include powders, tablets, pills, granules, dragees, hard or soft capsules, liquids, emulsions, It can be formulated in the form of suspensions, syrups, elixirs, external preparations, suppositories, sterile injectable solutions and the like orally or parenterally, particularly preferably orally.

Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules and the like, which solid dosage forms contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin in β-glucan. The mixture is prepared. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid dosage forms for oral administration include suspensions, solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used.

 Furthermore, pharmaceutical compositions comprising β-glucans of the present invention can be prepared using any suitable method known in the art or as disclosed in Remington's Pharmaceutical Science (Recent Edition, Mack Publishing Company, Easton PA). It may be preferably formulated using.

The dosage of β-glucan of the present invention may vary depending on the weight, age, sex, health condition, diet, time of administration, administration method, excretion rate, and severity of the disease, but β-glucan in the pharmaceutical composition. Or the effective dosage of derivatives thereof is typically 0.1 to 1000 mg / kg per day for adults (70 kg), can be administered 1 to several times a day . Since the dosage may vary depending on various conditions, it will be apparent to those skilled in the art that the dosage may be added or subtracted, and thus the dosage does not limit the scope of the present invention in any aspect.

The frequency of administration can be administered once a day or divided into several times within the desired range, the administration period is not particularly limited. In addition, the composition of the present invention may be ingested daily by adding to any food, in addition to oral administration as it is.

The pharmaceutical composition comprising β-glucan of the present invention may be administered to mammals such as mice, mice, livestock, humans, and the like by various routes.

The pharmaceutical composition comprising β-glucan or derivatives thereof of the present invention not only provides excellent liver disease treatment effect, but also has no toxicity and side effects due to the drug, so that it can be used with confidence even during long-term administration.

Accordingly, the β-glucan or derivatives thereof may be used as a main, subsidiary and food additive of other foods.

 Accordingly, the present invention provides a dietary supplement for liver function enhancement comprising the β-glucan described above.

According to an embodiment of the present invention, the food comprises a glucose main chain connected by β-1,3 bonds, and at least one of the glucose is branched glucose and gentiobiose connected by β-1,6 bonds. Phosphorus water-soluble β-1,3 / 1,6-glucan or derivatives thereof.

As used herein, the term "health functional food" is a food manufactured and processed using raw materials or ingredients having useful functions for the human body, and for health purposes such as regulating nutrients or physiological effects on the structure and function of the human body. Ingestion means to obtain a useful effect.

The health functional food containing the β-glucan or derivatives thereof may include drugs, foods and beverages in the form of tablets, capsules, powders, granules, liquids, and pills. Specifically, various foods, candy, chocolate, a drink, a gum, tea, a health supplement food, etc. are mentioned, for example.

In this case, the amount of β-glucan or derivatives thereof in the food or beverage may generally be about 0.01 to about 50% by weight, preferably about 0.1 to about 20% by weight of the total food weight. In the case of about 0.02 to about 10 g, preferably about 0.3 to about 1 g, based on 100 ml.

The dietary supplement may further comprise a food supplement acceptable food additive with the β-glucan or derivatives thereof.

The health beverage composition comprising the β-glucan or derivatives thereof has no particular limitation on the liquid component except for containing the β-glucan or derivatives thereof as essential ingredients in the indicated ratios, and various flavoring agents as in general beverages. Or natural carbohydrate or the like as an additional component. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like; And sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, baudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can also be used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to about 12 g per 100 ml of the health beverage composition.

In addition, various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, such as colorants and fillers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective properties And colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the nutraceutical may contain natural fruit juice and fruit flesh for the production of fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical but is generally selected from about 0.01 to about 20 parts by weight per 100 parts by weight of the health functional food.

Hereinafter, examples are provided to help understand the present invention. However, the following examples are provided only to more easily understand the present invention, and the present invention is not limited to the following examples.

Example  Β- used to treat liver disease Glucan  Produce

Β-glucan derived from Aureobasidium pullulans SM-2001 was prepared in powder as described in Korean Patent No. 0443209.

In summary, Aureobasidium pullulans SM-2001 (KCCM 10307) was inoculated into a culture medium containing glucose as a carbon source, followed by shaking culture at 30 ° C for 3 days. The culture solution was centrifuged to recover the supernatant, extracted with 95% ethanol and dialyzed, and then vacuum lyophilized to prepare a water-soluble β-1,3 / 1.6 glucan. The prepared glucan was stored at 4 ° C. to prevent light from being used until use.

Β-glucan from Aureobasidium pullulan SM-2001 is also commercially available (Glucan Corp., Korea).

Example  2 Preparation of animal model of liver disease and administration of test substance

Example  2-1 Preparation of Liver Disease Animal Model

To investigate the effects of β-glucan on liver disease, rat models of Streptozotocin (Sigma USA) (diabetic liver disease animal model) and CCl4 (subacute liver injury animal model) induced liver disease rats were prepared as follows. Feed and water were freely ingested during the day during the experiment. In addition, the light-dark cycle in the cage was adjusted to repeat day and night every 12 hours.

In the diabetic liver disease animal model (STZ rats), 100 female rats (6 weeks old, 120-140 g, SLC, Japan, delivered to the laboratory) were used in this experiment after adapting to the surrounding environment for 8 days. 88 of these animals were intraperitoneally administered Streptozotocin (STZ) dissolved in 50 mM citric acid buffer at a total concentration of 60 mg / kg / 5 ml. The remaining 12 animals were administered with the same volume of medium (water) in the same way instead of STZ for use as Sham controls. Furthermore, in order to measure the effect of β-glutan of the present invention according to the condition of the disease, rats induced by diabetes mellitus by STZ were divided into mild (L), moderate symptoms and severe (H) according to the severity.

For the subacute liver injury animal model (CCl4 rats), 56 female Sprague-Dawley rats (6 weeks old at delivery, 145-165 g, Samtako Bio, Korea) were used in this experiment after 10 days of adaptation to the surrounding environment. . CCl4 (Carbon tetrachloride, Deoksan Chemical, Korea) was administered subcutaneously to CCl4 mixed with olive oil at 7.92% v / v at a concentration of 0.15ml / kg once every two days for three weeks. The remaining eight animals were administered with the same volume of medium (water) in the same way instead of CCl4 for use as the Sham control.

Example  2-2 Administration of Test Substance

β- Glucan  administration

Β-glucan prepared according to Example 1 was diluted with distilled water in amounts of 31.25, 62.5 and 125 mg / kg in 2 ml of total volume. The β-glucan solution was then orally administered to rats prepared according to Example 2-1 using a 3 ml syringe with rat oral sonde: (1) For STZ rats, administration of STZ 25 From days, once daily for 4 weeks at concentrations of 62.5 and 125 mg / kg, respectively; And (2) for CCl4 rats, once daily for 42 days at 31.25, 62.5 and 125 mg / kg concentrations, respectively, at the same time as CCl4 administration.

Administration of known liver disease therapeutics as a control

As a positive control for the test for the treatment of liver disease of β-glucan, instead of β-glucan for STZ rats, SILY (Silymarin, Sigma, USA) (25 mg / kg / 5 ml), and CAPT (for CCl4 rats) Catopril, Sigma, USA) (100 mg / kg / 5 ml) and SILY (25 mg / kg / 5 ml) were each administered orally.

Example  Β- Using a Liver Disease Animal Model Glucan  Evaluation of liver disease treatment effect

Example  3-1 Effects on Diabetic Liver Disease

After administration of β-glucan to STZ rats prepared according to Example 2 as in Example 2-2, body weight, blood AST (Aspartate Transferase, GOT) and ALT (Alanine Transferase (GPT) content as described below Changes, and histological changes were observed.

All data are expressed as mean ± standard deviation. The number of individuals in each dosing group was 6, and statistical analysis was performed using SPSS (Release 6.1.3., SPSS Inc., USA) for Windows as Mann-Whitney U-Wilcoxon Rank Sum W test (MW test). For each of these variables, the percentage change (PCB) for the media control was PC vs media control (%) = [(data of test group-data of media control)] / data of media control] x 100; And percentage change (PCA) for the Sham control group was calculated from PC vs Sham control (%) = [(test group data-Sham data)] / Sham data] × 100.

Impact on weight change

Body weights were measured on the day of STZ administration, on the day of β-glucan administration, and at 7, 14, 21, 27 and 28 (rat sacrifice time), respectively. Before weighing, each animal was fasted overnight to reduce dietary errors.

The results are summarized in Table 1 below.

In the above table, the names of the respective administration groups are as follows: G0; Sham, normal media control; G1: Hepatopathogenic media control (β-glucan untreated group); G2: SILY 25 mg / kg dose group; G3: glucan 62.5 mg / kg / 5 ml administration group; G4: glucan 125 mg / kg 5 ml administration group; HC; Severe hepatic induction media control; HG; Glucan 62.5 mg / kg / 5 ml group after severe liver disease induction; LC; Mild hepatic induction medium control; LG; Glucan 62.5mg / kg / 5ml group after mild hepatic complication. Medium: 5 ml / kg sterile distilled water.

As shown in Table 1, during the induction period of diabetic liver disease, a significant (p <0.01 or p <0.05) weight loss (-70.13% compared to the Sham control group) was observed in all STZ-treated groups compared to the normal group. This suggests that effective liver disease has been caused. On the other hand, the weight gain during the period of drug administration was significantly decreased in the medium control group compared to the normal group, but the weight loss was remarkably recovered by the administration of β-glucan, and rats treated with β-glucan at 62.5 mg / kg were The weight gain was 383.78% compared to the control group, whereas the weight gain was -79.56% without β-glucan. This effect is superior to that of silymarin (weight gain of -15.31% for the media control group) previously known as a liver disease treatment agent, indicating the superiority of β-glucan as a treatment for liver disease.

On the other hand, there was no significant change in body weight in the GLUCAN-treated group in the severe diabetic and mild-induced group. This change was so severe that the weight loss caused by diabetes itself, rather than the weight loss caused by diabetic liver disease, was not observed.

Blood ALT  And AST Impact on

AST (Aspartate Transferase, GOT) and ALT (Alanine Transferase, GPT) are representative indicators of liver disease status and its extent. Rats were sacrificed 28 days after the administration of β-glucan, blood was collected, and plasma was isolated according to a known method. AST and ALT concentrations were measured from the separated plasma using an automated hematology analyzer (Toshiba 200 FR, Japan) based on the Kinetic UV method. The results are summarized in Table 2.

Abbreviations are as defined in Table 1. * P <0.01 compared to Sham control by MW test and #p <0.01 and ## p <0.05 compared to media control by MW test.

As shown in Table 2, the STZ administration group showed a significant increase in blood AST (502.83%) and ALT (974.67%) content compared to the normal group, indicating that the liver disease was effectively induced in rats. However, the increased AST and ALT concentrations were significantly reduced by up to about 50% in the β-glucan administration group of all concentrations. These blood levels of AST and ALT were significantly decreased in all liver diseases compared to the media control group not receiving β-glucan, regardless of disease severity. This effect is comparable to that of silymarin, previously known as a hepatic disease treatment agent, and exhibits superiority of β-glucan as a hepatic disease treatment agent.

Histopathological examination

Liver was harvested at the end of the administration of each group to which β-glucan was administered to STZ rats and STZ rats. The extracted liver was fixed in formalin buffered at a neutral pH of 10%, embedded in paraffin according to a known method, cut into sections having a thickness of 3 to 4 μm, stained with Hematoxylin and Eosin (H & E), and then subjected to optical microscopy. Observed. The results are in FIGS. 1 and 2.

As shown in FIG. 1, STZ rats ((1c), (1d) and (1e)) to which β-glucan was administered showed histological findings close to that of normal rats compared to rats (1b) not administered. Seemed. Particularly, as shown in FIG. 2, liver fibrosis was observed in HC group (a, b), which is a severe liver disease causing group, but such fibrosis was not observed in HG group (c) to which β-glucan was administered. It has been demonstrated that β-glucan has an excellent effect on liver disease.

Taken together, the β-glucan of the present invention can be dose-dependently reduced in blood AST and ALT content, which is a direct indicator of weight, histopathological changes, especially liver status. It is thought to be very effective for liver disease including liver disease.

Example  3-2 Effects on Subacute Liver Disease

 After administration of β-glucan to CCl4 rats prepared according to Example 2 as in Example 2-2, the body weight, blood AST (Aspartate Transferase, GOT) and ALT in the same manner as in Example 3-1, unless otherwise stated (Alanine Transferase (GPT) content change and histological changes were observed to examine the effect of β-glucan on subacute liver disease. The results are shown in the following Tables 3 to 6 and FIG. 3.

Impact on weight change

Body weights were measured one day before the start of β-glucan administration and on days 7, 14, 21, 28, 35 and 41 and 42 (rat sacrifice time), respectively. Before weighing, each animal was fasted overnight to reduce dietary errors.

The results are summarized in Table 3 below.

1) Initial administration after overnight fasting, 2) Body weight at sacrifice after overnight fasting, 3) Weight gain (g) is increase in body weight from day 0 to day 42, and 4) PC-S, Sham (G0) Percent change in body weight; 5) Weight change percentage (%) for PC-C, media control; * P <0.01 and ** p <0.05 compared to Sham control by MW test.

In the above table, the names of the respective administration groups are as follows: Sham: normal medium control; Control: Hepatopathogenic media control; CAPT: Captopril 100 mg / kg dose group; G31.25, G62.5, G125: 32.5 mg / kg / 5 ml, 62.5 mg / kg / 5 ml, and 125 mg / kg / 5 ml β-glucan treated groups, respectively; SILY: Silymarin 25 mg / kg dose group; Medium: 5 ml / kg sterile distilled water.

As shown in Table 3, significant (p <0.01) weight loss was observed in the subacute liver disease rats treated with the medium control group, suggesting that effective liver disease was induced. However, this weight loss was recovered dose-dependently when β-glucan was administered, resulting in up to 15.27% weight gain in rats administered at 125.5 mg / kg compared to the control group, while β-glucan was not administered. If not, the weight gain was -38.08%. This effect is far superior to Captopril (CAPT) and Silymarin (SILY) (weight gain for the media control, respectively -3.05 and -1.63%, respectively), previously known as liver disease treatment agents. It indicates excellence.

Blood ALT  And AST Impact on

AST and ALT were measured in the same manner as in SZT rats except that the rats were sacrificed 42 days after the administration of β-glucan. The results are summarized in Table 4.

Abbreviations are as defined in Table 3. Plasma AST and ALT concentrations are the plasma concentrations at sacrifice 42 days after the administration of the test compound comprising β-glucan, minus the plasma concentrations at the beginning of the compound administration. * P <0.01 compared to Sham control by MW test and #p <0.01 and ## p <0.05 compared to media control by MW test.

As shown in Table 4, the STZ administration group showed a significant increase in blood AST (727.80%) and ALT (1327.78%) content compared to the normal group, indicating that the liver disease was effectively induced in CCl4 rats. However, the increased AST and ALT concentrations showed a concentration-dependent decrease in the β-glucan administration group, resulting in a significant decrease of up to 50% at a concentration of 125 mg / kg. These effects include captopril (CAPT, -20.95 and -43.42% for AST and ALT, respectively) and silymarin (-29.12 and -36.51%, respectively), previously known as liver disease agents. As a superior effect, it shows the superiority of β-glucan as a therapeutic agent for liver disease.

Histopathological examination

Histological analysis of the liver was performed in the same manner as described in Example 3-1. Furthermore, in this example, the degenerative hepatic region of the hepatic parenchyma was measured by histomorphometry, which can more directly identify liver damage. The liver tissues prepared as described above were measured using a microscope equipped with an automaticSIS image processing (ANAlySIS Image Processing; SIS, Germany) to measure the percentage of necrotic cells per field (200 μm ² ). The results are shown in Table 5 and FIG. 3.

Abbreviations are as defined in Table 3.

As shown in Table 5, CCl4 rats that did not receive β-glucan had an increase of 1189.69% cell necrosis in hepatic parenchyma compared to the medium control group, whereas the β-glucan group had a maximum dose of -42.49 at a concentration of 125 mg / kg. A significant decrease in% was shown. This is far superior to Captopril (CAPT) and Silymarin (SILY), previously known to treat liver disease (-21.13 and -37.91%, respectively). In addition, these results are consistent with the histomorphologic analysis of FIG. 3, as shown in FIG. 3, while the media control group showed inflammatory cell infiltration with severe cell necrosis, with hepatic fibrosis in the central lobules 2. , β-glucan treated groups ((5), (6) and (7)) show concentration-dependent relief of these symptoms, comparable to or better than the effects of captopril (3) and silymarin (4). will be.

These results are consistent with the effect of β-glucan on weight gain and AST and ALT concentrations, demonstrating the superiority of β-glucan as a therapeutic agent for various liver diseases of the present invention. Furthermore, no abnormal syndrome (hair loss, etc.) was observed in the animal model during the experimental period, and it was confirmed that β-glucan was not particularly toxic to the animal model.

Taken together, all of the above results indicate that β-glucan of the present invention has an excellent effect on diabetic liver disease as well as subacute liver disease, and thus, β-glucan of the present invention is a therapeutic agent for various liver diseases. It proves that it can be used effectively.

The β-1,3 / 1,6-glucan prepared in Example 1 may be administered in the following formulations, and the following formulations are merely illustrative of the present invention, thereby not limiting the contents of the present invention.

Formulation Example 1 Preparation of Tablet

200 mg of β-1,3 / 1,6-glucan of Example 1

Lactose 100mg

Starch 100mg

Magnesium stearate proper amount

Tablets were prepared by mixing and tableting the above components according to a conventional tablet production method.

Formulation Example 2 Preparation of Capsule

 Β-1,3 / 1,6-glucan 100 mg of Example 1

Lactose 50mg

Starch 50mg

Talc 2mg

Magnesium stearate proper amount

According to the conventional capsule preparation method, the above ingredients were mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 3 Preparation of Liquid

Β-1,3 / 1,6-glucan 1000 mg of Example 1

20 mg of sugar

Isomerized sugar 20mg

Lemon flavor

Purified water was added to make a total of 1000 ml. According to the conventional method for preparing a liquid, the above components were mixed, and then filled into a brown bottle and sterilized to prepare a liquid.

Formulation Example 4 Preparation of Injection

 Β-1,3 / 1,6-glucan 100 mg of Example 1

Sodium metabisulfite 3.0mg

Methylparaben 0.8mg

Propylparaben 0.1mg

Proper sterile distilled water for injection

The above components were mixed and prepared in a conventional manner so that the final volume was 2 ml, and then filled into 2 ml ampoules and sterilized to prepare an injection.

Formulation Example 5 Preparation of Healthy Drink

 Β-1,3 / 1,6-glucan 1000 mg of Example 1

Citric acid 1000mg

Oligosaccharide 100g

Mesyl Concentrate 2g

* 163 g of taurine

Add 900 ml of purified water

After mixing the above components in accordance with a conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2L container, sealed sterilized and then refrigerated and stored in the present invention Used to prepare healthy beverage compositions.

Formulation Example 6 Preparation of Healthy Food

 Β-1,3 / 1,6-glucan 1000 mg of Example 1

Vitamin mixture proper amount

Vitamin A Acetate 70μg

Vitamin E 1.0mg

Vitamin B1 0.13mg

Vitamin B2 0.15mg

Vitamin B6 0.5mg

0.2 μg of vitamin B12

Vitamin C 10mg

Biotin 10μg

Nicotinic Acid 1.7mg

Folic acid 50μg

Calcium Pantothenate 0.5mg

Vitamin E 1.0mg

Mineral mixture

Ferrous Sulfate 1.75mg

Zinc Oxide 0.82mg

Magnesium Carbonate 25.3mg

15 mg potassium monophosphate

Dicalcium Phosphate 55mg

Potassium Citrate 90mg

Calcium Carbonate 100mg

Magnesium chloride 24.8mg

The composition ratio of the above-mentioned vitamin and mineral mixtures is composed of relatively suitable ingredients suitable for health foods in a preferred manufacturing example, but the compounding ratios may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. Health foods of granule formulations were prepared.

The composition comprising β-glucan of the present invention has superior therapeutic effect to diabetic liver disease and various liver diseases including subacute liver disease, and is a natural substance, orally administered as compared to other synthetic drugs. Of course, since the immunogenicity and side effects are extremely low, it can be used in place of conventional liver disease treatments that are problematic in the method of administration, side effects, safety and cost.

Claims (9)

Pharmaceutical composition for the treatment of diabetic liver disease, liver disease caused by subacute liver injury, or toxic liver disease caused by alcohol or compound, etc. comprising β-glucan as an active ingredient. The method of claim 1, wherein the β-glucan comprises a glucose main chain connected by β-1,3 bonds, and at least one of the glucose is β-1,3 / 1 connected by branched glucose to β-1,6 bonds. , 6-glucan. The composition of claim 2, wherein the β-glucan is β-1,3 / 1,6 glucan of formula (I): [Formula 1]

Wherein n is from 1 to about 4,000 and X is lactic acid, maleic acid or sulfoacetic acid. The composition according to any one of claims 1 to 3, wherein the β-glucan is derived from Aureobasidium pullulans SM2001. delete Health functional food for liver function enhancement comprising β-glucan as an active ingredient. The method of claim 6, wherein the β-glucan comprises a glucose main chain connected by β-1,3 bond, wherein at least one of the glucose is β-1,3 / 1 linked by branched glucose and β-1,6 bond Health functional food for liver function enhancement, characterized in that 6-glucan. [Claim 9] The health functional food for liver function enhancement of claim 7, wherein the β-glucan is β-1,3 / 1,6 glucan of the following Chemical Formula 1. [Formula 1]

Wherein n is from 1 to about 4,000 and X is lactic acid, maleic acid or sulfoacetic acid. The health functional food for improving liver function according to any one of claims 6 to 8, wherein the β-glucan is derived from Aureobasidium pullulans SM2001.

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