KR101158472B1 - Composition for Preventing or Treating of Alcoholic Liver Disease - Google Patents

Abstract

The present invention relates to a composition for preventing or treating liver disease caused by an alcohol comprising a compound represented by the following general formula (I) as an active ingredient:

Formula I

S- (MS) _p- (MS) _q -H

In the above general formula, S is sialic acid, (MS) _p and (MS) _q are independently of each other monosaccharide residues.

The compound used as an active ingredient of the present invention results in a decrease in GOT and GPT levels in the alcohol dietary model, ultimately exhibiting the prophylactic or therapeutic activity of alcohol-induced liver disease. In addition, the compositions of the present invention can be safely applied to pharmaceutical compositions, neutraceutical compositions or food compositions.

Sialic acid, sialyl lactose, alcohol, liver disease, fatty liver

Description

Composition for Preventing or Treating Alcoholic Liver Disease

The present invention relates to a composition for preventing or treating liver disease caused by alcohol.

The liver is the largest organ in our body and is a very important organ responsible for various metabolism, detoxification, decomposition, synthesis and secretion. Looking more closely, first, it has the function of managing energy metabolism, so that all the nutrients absorbed from foods are metabolized into substances that can produce energy in the liver and are supplied or stored throughout the body. Second, the liver has a function of synthesizing, storing, and distributing about 2,000 kinds of enzymes, albumin, and serum proteins such as coagulation factors, bile acids, phospholipids, or cholesterol. Third, as the detoxification and decomposition functions detoxify drugs, alcohol, or toxic substances, hepatic cells are easily damaged, and therefore drug, toxic or alcoholic liver diseases are frequently generated. In addition, the metabolites are excreted in the duodenum through the bile ducts and the immune function plays an important role in maintaining our lives.

One of the main causes of liver disease is alcohol. Hepatic diseases caused by alcohol are classified as alcoholic fatty liver, alcoholic hepatitis, and alcoholic cirrhosis, and take many forms, ranging from accumulation of some fat in the liver to advanced cirrhosis. Citing data from the Korea National Statistical Office in 2007, the prevalence of alcoholic liver disease shows that the number of deaths from alcohol-related diseases in Korea is 9.6 per 100,000 population, which has a significant impact on alcohol health. The cause of cirrhosis in Korea is the second place cirrhosis caused by drinking alcohol, and drinking is the third place in the case of hepatocellular carcinoma in Korea. Alcoholic fatty liver is found in 80% of heavy drinkers, 15-20% experience hepatitis, and 10% progress to alcoholic cirrhosis (1, 2).

Alcoholic fatty liver

Fatty liver is caused by alcohol, obesity, and diabetes, and the accumulation of triglycerides in liver cells. Reversible liver disease can be recovered by removing the cause. If you drink 80 grams of alcohol daily, fatty liver is found in almost all cases. Lipid droplets make up the bulk of the cell, while liver function levels are still normal. If you drink alcohol in this state, you can return to normal, but if you continue drinking, go to cirrhosis.

Acute Alcoholic Hepatitis

Hepatitis is a disease in which hepatocytes are destroyed for a short time or for a long time due to viruses, drugs, or toxic substances. In chronic active hepatitis, the virus proliferates actively, and the hepatocytes are continuously destroyed to progress to cirrhosis for a long time. As alcoholic hepatitis progresses, hepatocytes grow as the water content increases in the cells. Other biopsy findings include balloon or feather degeneration and neutrophil infiltration. Alcohol hepatitis occurs after drinking alcohol for at least 15-20 years. In other words, absolute drinking volume and total drinking duration are factors that determine hepatitis, and drinking habits and types of alcohol do not matter. Symptoms include severe jaundice and nonspecific systemic symptoms, with a high mortality rate of 30-60%. At this stage, even if this week, the clinical course may be worse.

Alcoholic cirrhosis

Liver cirrhosis is caused by long-term, persistent liver damage caused by chronic hepatitis or alcoholic liver disease. When cirrhosis occurs, hepatic function is markedly impaired and blood flow through the liver is impaired. Ascites, gastrointestinal bleeding, hepatic Complications such as hepatic coma, esophageal varices, spontaneous peritonitis, or hepatoenal syndrome are common. Liver cirrhosis is the final stage of alcoholic liver disease, initially showing sintered nodule (<3 mm) cirrhosis, but changes over time or due to insufficient nutrition to colonic cirrhosis. It is more common in persistent drinkers than in those who occasionally have a habit of binge drinking. Fibrosis develops around the cell, around the Disse cavity, or around the central vein. Central hyaline sclerosis findings herald progression to cirrhosis and are more often observed in female drinkers. The fibrous bridge (collagen bridge) is connected between the central vein and the portal vein. When several bridges are connected, hepatocellular masses are isolated by bands and form a regenerative nodule. The 1-year survival rate is 60-70% and the 2-year survival rate is 35-50%. Liver cancer can occur in people who have chronic hepatitis virus or have suffered from cirrhosis for a long time. Treatment is often difficult because of severe progression or severe liver failure. Currently, liver fibrosis is the leading cause of death from liver disease. In Western society, 50% of cirrhosis diseases are caused by excessive alcohol intake, and 83% of men are over 20 years old in Korea. In addition, in Korea, the drinking population of women is rapidly increasing, so it is urgent to research and develop substances for preventing and treating alcoholic liver disease (3).

Alcohol Fatty liver alone does not require special treatment except alcohol. Alcoholic hepatitis has a number of treatments, including steroids, antioxidants, nutritional supplements, pentoxifylline or anti-TNF antibodies. Patients with non-alcoholic cirrhosis who have been treated with propylthiouracil, colchicine, antioxidants or phosphatidylcholine as drug therapy in addition to this week have not yet been proven effective, and liver transplant therapy is the only option for low life expectancy. Means (4-6).

Throughout this specification many patent documents are referenced and their citations are indicated. The disclosures of the cited patent documents are hereby incorporated by reference herein in their entirety to better illustrate the state of the art to which the invention pertains and the teachings of the present invention.

The present inventors have tried to develop a substance for preventing or treating alcoholic liver disease. As a result, the present invention was completed by confirming that sialyl oligosaccharides reduce the GOT and GPT levels in the alcohol dietary model, thereby improving the liver disease caused by alcohol.

Accordingly, it is an object of the present invention to provide a composition for preventing or treating liver disease caused by alcohol.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to an aspect of the present invention, the present invention provides a composition for preventing or treating liver disease caused by an alcohol comprising a compound represented by the following general formula I as an active ingredient:

Formula I

S- (MS) _p- (MS) _q

In the general formula, S is sialic acid, (MS) p and (MS) q are independently monosaccharide residues.

The present inventors have tried to develop a substance for preventing or treating alcoholic liver disease. As a result, it was confirmed that sialyl oligosaccharides reduce the GOT and GPT levels in the alcohol dietary model, thereby improving the alcohol-induced liver disease.

In the present invention, the active ingredient is a compound of formula (I). In Formula I, S represents sialic acid. Sialic acid may be bound to (MS) _p in a variety of ways, but most preferably to a monosaccharide compound (MS) _p with an α2,3 bond. In addition to sialic acid, S may be modified sialic acid. For example, a modification of the —OH group at carbon 4 of sialic acid (eg, modified by a C ₁ -C ₄ alkyl group) may be located at S. Most preferably, unmodified sialic acid is located in S.

The monosaccharide compounds located at (MS) _p and (MS) _q can use any monosaccharide compound known in the art, for example, pentose (eg erythrose and threose), pentose (eg ribose) , Arabinose, xylose and lysoose) and hexasaccharides (allose, altrose, glucose, mannose, gulose, iodos, galactose and thalos). Preferably, the monosaccharide compound located at (MS) _p and (MS) _q is pentose or hexasaccharide, more preferably hexasaccharide, even more preferably glucose, mannose or galactose, most preferably Is glucose or galactose. The monosaccharide compounds located at (MS) _p and (MS) _q may have a stereoisomer of D- or L-form, most preferably a stereoisomer of D-form.

The same or different monosaccharide compounds may be bonded to (MS) _p and (MS) _q , and preferably different monosaccharide compounds are bonded.

According to a preferred embodiment of the present invention, (MS) _p is combined with galactose or glucose, (MS) _q is combined with glucose or galactose, and most preferably (MS) _p is combined with galactose and (MS) _q with glucose. . Galactose is bonded to (MS) _p and glucose is bound to (MS) _q , and a disaccharide compound and lactose are formed.

Monosaccharide compounds located at (MS) _p and (MS) _q are modified or unmodified. For example, in the case of modified monosaccharide compounds, an acetyl group or an N-acetyl group may be bonded to the -OH group. Preferably, the monosaccharide compounds located at (MS) _p and (MS) _q are unmodified monosaccharide compounds.

The most preferred embodiment of the compound of formula I used as an active ingredient in the present invention is sialic acid-galactose-glucose. Sialic acid-galactose-glucose is sialylactose.

Sialic acid can be bound to galactose in a variety of ways, such as by α2,3 or α2,6 bonds. Sialic acid may be modified, for example the —OH group at carbon 4 of sialic acid may be modified (eg, modified by a C ₁ -C ₄ alkyl group).

Galactose and glucose in sialylactose may have D- or L-form stereoisomers and most preferably are D-form stereoisomers. Galactose and glucose in sialylactose are modified or unmodified. For example, in the case of modified monosaccharide compounds, an acetyl group or an N-acetyl group may be bonded to the -OH group. Preferably, galactose and glucose in sialylactose are unmodified monosaccharide compounds.

According to a preferred embodiment of the present invention, sialyl lactose used as an active ingredient in the present invention is α-NeuNAc- (2 → 3) -β-D-Gal- (1 → 4) -D-Glc or α-NeuNAc -(2 → 6) -β-D-Gal- (1 → 4) -D-Glc [NeuNAc: N-Acetylneuraminyl, Gal: Galactose, Glc: Glucose]. α-NeuNAc- (2 → 3) -β-D-Gal- (1 → 4) -D-Glc is a substance found in GM3 gangliosides and α-NeuNAc- (2 → 6) -β-D- Gal- (1 → 4) -D-Glc is an isomer of this material.

More preferably, sialylactose used as an active ingredient in the present invention is α-NeuNAc- (2 → 6) -β-D-Gal- (1 → 4) -D-Glc. As demonstrated in the examples below, α-NeuNAc- (2 → 6) -β-D-Gal- (1 → 4) -D-Glc is α-NeuNAc- (2 → 3) -β-D- Reduction of GOT and GPT levels is better in alcohol diet models than Gal- (1 → 4) -D-Glc.

According to a preferred embodiment of the present invention, the compound represented by the general formula (I) used as an active ingredient of the present invention is a glutamic-oxaloacetic transaminase (GOT) or article of serum Reduce levels of glutamic-pyruvic transaminase (GPT).

As used herein, the term “glutamic-oxaloacetic transaminase (GOT)” is an indicator of diagnosing liver injury, and the GOT is an enzyme generally present in liver and heart cells, Promote the reversible transfer of the amino groups of L-glutamic acid and oxaloacetic acid to form α-ketoglutaric acid and L-aspartic acid. When the liver is damaged by various causes (eg, alcohol, stress or hepatitis virus, etc.), the enzyme is released into the blood, resulting in increased levels of the enzyme in the serum. It is also called aspartate aminotransaminase (AST) or glutamic-aspartic transaminase.

As used herein, the term “glutamic-pyruvic transaminase (GPT)” is an indicator of liver injury diagnosis, such as GOT, and is an enzyme found in serum and various body tissues, but generally It is the enzyme most closely associated with the liver. The enzyme is an enzyme that promotes the transfer of amino groups of alanine and α-ketoglutarate, and forms pyruvate and glutamate. When GPT is also damaged, the enzyme is released into the blood, which increases the level of the enzyme in the serum. And this is called alanine transaminase (ALT) or alanine aminotransferase (ALAT).

The composition of the present invention comprising the above-described compound of Formula I as an active ingredient results in a reduction of GOT and GPT levels in the alcohol dietary model, ultimately showing the prophylactic or therapeutic activity of alcohol-induced liver disease.

According to a preferred embodiment of the present invention, the liver disease caused by alcohol prevented or treated by the composition of the present invention is alcoholic fatty liver, alcoholic hepatitis, alcoholic chronic hepatitis, alcoholic liver fibrosis, alcoholic cirrhosis or alcoholic liver cancer, more preferred Alcoholic fatty liver, alcoholic hepatitis, alcoholic liver fibrosis or alcoholic cirrhosis.

The composition of the present invention may be prepared as a pharmaceutical composition, a neutraceutical composition or a food composition.

According to a preferred embodiment of the present invention, the composition of the present invention comprises (a) a pharmaceutically effective amount of the compound of formula I of the present invention as described above; And (b) a pharmaceutically acceptable carrier.

As used herein, the term “pharmaceutically effective amount” means an amount sufficient to achieve the efficacy or activity of the compounds of Formula I described above.

When the composition of the present invention is made into a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington &apos; s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, mucosal administration, and eyedrop administration.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . Preferably, the dosage of the pharmaceutical composition of the present invention is 0.001-100 mg / kg body weight.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or it may be prepared by incorporation into a multi-dose container. The formulation may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of extracts, powders, powders, granules, tablets or capsules, and may further comprise dispersants or stabilizers.

When the composition of the present invention is prepared from a food composition (or functional food composition), it contains not only the compound of general formula (I) as an active ingredient but also components commonly added during food production, and include, for example, proteins, carbohydrates, Fats, nutrients, seasonings and flavoring agents. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

 For example, when the food composition of the present invention is prepared with a drink, in addition to the compound of formula I of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, etc. Can be included.

As described above in detail, the present invention provides a composition for the prevention or treatment of liver disease caused by an alcohol comprising a compound represented by the general formula (I) described above as an active ingredient. The compound used as an active ingredient of the present invention results in a decrease in GOT and GPT levels in the alcohol dietary model, ultimately exhibiting the prophylactic or therapeutic activity of alcohol-induced liver disease. In addition, the compositions of the present invention can be safely applied to pharmaceutical compositions, neutraceutical compositions or food compositions.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Experimental Materials and Experimental Methods

Experimental material

Experimental animals were purchased from Dooyeol Biotech (Korea) with 6-week-old female C57BL / 6 mice and normal pellet feed. The lieber-DeCarli ethanol diet for the ethanol diet was purchased from Dyets (USA) and 100% ethanol was purchased from Sigma-Aldrich (USA). GOP / GPT assay kit for measuring liver disease value was purchased from Asan Pharmaceutical Co., Ltd. (AM101-K, South Korea).

Induction of Alcoholic Liver Disease

Six-week-old female C57BL / 6 mice were allowed to acclimate for one week while feeding on normal feed. Mice were divided into four groups as follows and ethanol feed was fed to induce alcoholic liver disease: normal group of mice fed normal diet; Group of ethanol dietary mice fed control lieber-DeCarli ethanol feed; Experimental group A group of mice intraperitoneally administered 3'- or 6'-sialylactose in amounts of 0.01 mg / kg and 0.1 mg / kg to mice fed lieber-DeCarli ethanol feed. Ethanol feed was fed by gradual increase of ethanol content at weekly intervals so that 9%, 18%, 27% or 36% of total calories were fed from ethanol. Each group consisted of four mice each.

Therapeutic Effect of Alcoholic Liver Disease

The therapeutic effect of alcoholic liver disease was confirmed by injecting 3'- or 6'- sialylactose daily into the abdominal cavity of lieber-DeCarli ethanol feed for 4 weeks in amounts of 0.01 mg / kg and 0.1 mg / kg. . Control rats were intraperitoneally injected with phosphate buffer solution.

Confirmation of Alcoholic Liver Disease Treatment Effect

In order to confirm the therapeutic effect of alcoholic liver disease, blood GOT (Glutamic Oxaloacetic Transaminase) and GPT (Glutamic Pyruvic Transaminase) levels were measured. GOP / GPT is present in the myocardium, liver, skeletal muscle and kidneys, but very small amounts in the blood. Thus, elevated levels reflect cellular degeneration and necrosis of organs, particularly as a potent indicator of liver damage. After the experiment, mice were sacrificed and whole blood was collected from the heart. After coagulation, the blood was centrifuged at 13,000 rpm for 20 minutes to obtain serum and stored frozen until analysis. Serum GOP / GPT was measured using Asan Pharmaceutical's GOP / GPT analysis kit (AM101-K).

Experiment result

6’- Sialylactose  Liver disease treatment effect

In the control group that had been given alcohol diet for 4 weeks, GOT level was increased by 258% compared to normal group and GPT level was increased by 513%. On the other hand, the experimental group intraperitoneally administered 6′-sialylactose in an amount of 0.1 mg / kg showed 32.0% GOT and 51.2% GPT. GOT and GPT levels are major indicators of liver disease, and thus 6′-sialylactose has the effect of improving liver disease caused by alcohol. The experimental results are summarized in FIGS. 1 and 2.

3’- Sialylactose  Liver disease treatment effect

In the control group that had been given alcohol diet for 4 weeks, GOT level was increased by 145% compared to normal group, and GPT level was increased by 170%. In contrast, the experimental group intraperitoneally administered 3′-sialylactose in an amount of 0.1 mg / kg showed 60.3% of GOT and 41.2% of GPT. GOT and GPT levels are major indicators of liver disease, and thus 3′-sialylactose has the effect of improving liver disease caused by alcohol. The experimental results are summarized in FIGS. 3 and 4.

Based on the experimental results, sialyl lactose is effective in reducing alcoholic liver disease by reducing the level of GOT or GPT, and in particular, the effect of 6'-sialylactose is better than that of 3'-sialylactose. Lilactose can prevent or treat alcoholic liver disease.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

references

1.Kim JR, ed. Disease of the digestive system. 1 ^st ed. Seoul: Ilchogak, 2000: 578-583

Chae HB Korean J Gastroenterol 53; 275-283 (2009)

3.Anland BS West J Med 171; 11-115 (1999)

4.Akriviadis E et al, Gastroenterology 119; 1637-1648 (2000)

Cabre E et al, Hepatology 32; 36-42 (2000)

6. Greenwell P. Alcohol. Clin. Exp. Res., 23930-933 (1999)

1 is a graph showing changes in GOT levels in alcohol diet C57BL / 6 mice administered 6′-sialylactose. Normal Diet, Et-OH Diet, and Et-OH + 6'SL were supplemented with 6'-siaallylactose (SL) in normal diet mouse group (normal group), alcohol diet mouse group (control group) and alcohol diet group, respectively. The experimental group treated.

FIG. 2 is a graph showing changes in GPT levels in alcohol diet C57BL / 6 mice administered 6′-sialylactose. Normal Diet, Et-OH Diet, and Et-OH + 6'SL were supplemented with 6'-siaallylactose (SL) in normal diet mouse group (normal group), alcohol diet mouse group (control group) and alcohol diet group, respectively. The experimental group treated.

3 is a graph showing changes in GOT values in alcohol diet C57BL / 6 mice administered 3′-sialylactose. Normal Diet, Et-OH Diet, and Et-OH + 3'SL were used to control 3'-sialylactose (SL) to normal diet mouse group (normal group), alcohol diet mouse group (control group) and alcohol diet group, respectively. The experimental group treated.

FIG. 4 is a graph showing changes in GPT levels in alcohol diet C57BL / 6 mice administered 3′-sialylactose. Normal Diet, Et-OH Diet, and Et-OH + 3'SL were used to control 3'-sialylactose (SL) to normal diet mouse group (normal group), alcohol diet mouse group (control group) and alcohol diet group, respectively. The experimental group treated.

Claims (12)

A pharmaceutical composition for preventing or treating liver disease caused by an alcohol containing sialyl lactose represented by the following general formula I as an active ingredient: Formula I S- (MS) _p- (MS) _q In the general formula, S is sialic acid, (MS) p is galactose, (MS) q is a composition, characterized in that glucose. delete delete The method of claim 1, wherein the sialyl lactose is α-NeuNAc- (2 → 3) -β-D-Gal- (1 → 4) -D-Glc or α-NeuNAc- (2 → 6) -β-D -Gal- (1 → 4) -D-Glc. The composition of claim 1, wherein the sialylactose is α-NeuNAc- (2 → 6) -β-D-Gal- (1 → 4) -D-Glc. The method of claim 1, wherein the sialyl lactose is glutamic-oxaloacetic transaminase (GOT) or glutamic-pyruvic transaminase (GPT) of serum A composition characterized by reducing the value of. The composition of claim 1, wherein the liver disease is alcoholic fatty liver, alcoholic hepatitis, alcoholic chronic hepatitis, alcoholic liver fibrosis, alcoholic cirrhosis, or alcoholic liver cancer. Functional food (neutraceutical) or food composition for the prevention or improvement of liver disease caused by alcohol containing sialyl lactose represented by the following general formula (I) as an active ingredient: Formula I S- (MS) p- (MS) q In the general formula, S is sialic acid, (MS) p is galactose, (MS) q is a composition, characterized in that glucose. The method of claim 8, wherein the sialyl lactose is α-NeuNAc- (2 → 3) -β-D-Gal- (1 → 4) -D-Glc or α-NeuNAc- (2 → 6) -β-D -Gal- (1 → 4) -D-Glc. The composition of claim 8, wherein the sialylactose is α-NeuNAc- (2 → 6) -β-D-Gal- (1 → 4) -D-Glc. The method of claim 8, wherein the sialyl lactose is glutamic-oxaloacetic transaminase (GOT) or glutamic-pyruvic transaminase (GPT) of serum A composition characterized by reducing the value of. 9. The composition of claim 8, wherein the liver disease is alcoholic fatty liver, alcoholic hepatitis, alcoholic chronic hepatitis, alcoholic liver fibrosis, alcoholic cirrhosis, or alcoholic liver cancer.

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