JP2008297204A - Hepatic fibrosis-inhibiting or improving agent containing polylactic acid mixture - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new hepatic fibrosis-inhibiting or improving agent by evaluating a hepatic fibrosis-inhibiting action exhibited by a cyclic and/or linear polylactic acid mixture. <P>SOLUTION: This hepatic fibrosis-inhibiting or improving agent contains a cyclic and/or linear polylactic acid mixture having a condensation degree of 3 to 20. The new hepatic fibrosis-inhibiting or improving agent, and foods and drinks using the hepatic fibrosis-inhibiting or improving agent can be provided by the present invention. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to an agent for suppressing or improving liver fibrosis, and more particularly to an agent for suppressing or improving liver fibrosis including a cyclic and / or chain polylactic acid mixture. The agent for suppressing or improving liver fibrosis of the present invention is useful for the treatment or prevention of liver diseases associated with liver fibrosis.

  The liver is an important organ with various functions such as detoxification, metabolism, and substance storage. However, it may suffer acute or chronic damage due to various causes such as viruses, drugs or alcohol. These cause diseases such as viral hepatitis, drug-induced liver injury, alcoholic liver injury, fatty liver, steatohepatitis, and cirrhosis and liver cancer. In addition to diet therapy and rest therapy, administration of glycyrrhizin preparations, corticosteroids or interferon, etc. are used as treatments for such liver diseases, but the effects of these treatments are not sufficient. Furthermore, since glycyrrhizin preparations and interferons are administered intravenously, they are not suitable for long-term administration. Furthermore, side effects are a problem with interferons and steroids.

  In Japan, chronic liver disease is one of the main causes of death, and in many cases, complications such as cirrhosis or liver cancer cause death. Many of the causes of chronic liver disease in Japan are viral hepatitis. In the case of chronic hepatitis due to hepatitis B virus or hepatitis C virus, inflammatory necrosis, expressed as fragmentary necrosis around the portal vein, is the most typical histological finding. After necrotic inflammation has begun, the limit membrane of hepatocytes around the portal vein is broken, inflammatory necrosis spreads to the liver, the portal vein is expanded, and inflammatory lesions cause the portal vein and portal vein to Is connected. Such necrotic inflammation, combined with progressive liver fibrosis, eventually progresses to cirrhosis.

  Liver fibrosis is a symptom that results from chronic liver damage induced by various causes. As liver fibrosis progresses, it eventually reaches cirrhosis. It is also known that when the degree of liver fibrosis increases, the incidence of hepatocellular carcinoma increases. Therefore, in order to prevent cirrhosis and hepatocellular carcinoma or prevent recurrence of hepatocellular carcinoma, it is important to suppress the progression of liver fibrosis.

  Previous studies have reported that cyclic and / or chain poly L-lactic acid mixtures having a condensation degree of 3 to 20 are useful as antineoplastic agents (Japanese Patent Application Laid-Open No. 9-227388 and special patents). (Kaihei 10-130153). However, there has been no report on the effect of cyclic and / or chain polylactic acid mixtures having a condensation degree of 3 to 20 on liver fibrosis.

JP-A-9-227388 JP-A-10-130153

  This invention made it the problem which should be solved to provide the novel inhibitor of liver fibrosis, or the improvement agent by evaluating the inhibitory action of hepatic fibrosis which the cyclic | annular and / or chain | strand-shaped polylactic acid mixture shows. This invention also made it the subject which should be solved to provide the food / beverage products using the suppression or improvement agent of above-mentioned liver fibrosis.

  As a result of intensive studies to solve the above problems, the present inventors have evaluated the effect of inhibiting liver fibrosis using a polylactic acid mixture. As a result, cyclic and / or chain polylactic acid mixtures having a condensation degree of 3 to 20 are obtained. It was demonstrated that hepatic fibrosis was suppressed. The present invention has been completed based on these findings.

That is, according to the present invention, there is provided an agent for suppressing or improving liver fibrosis, comprising a cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20.
Preferably, lactic acid which is a repeating unit in polylactic acid substantially consists of L-lactic acid.
Preferably, the agent for suppressing or improving liver fibrosis according to the present invention can be used for the treatment or prevention of liver damage associated with liver fibrosis.
According to another aspect of the present invention, there is provided a food or drink containing the above-described liver fibrosis inhibition or ameliorating agent of the present invention.

According to still another aspect of the present invention, there is provided use of a cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20 in the manufacture of an agent for suppressing or improving liver fibrosis.
According to still another aspect of the present invention, liver fibrosis is suppressed or improved, comprising administering an effective amount of a cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20 to a mammal such as a human. A method for doing so is provided.

  According to the present invention, it is possible to provide a novel inhibitor or improver of liver fibrosis, and a food or drink using the same. In addition, since the polylactic acid mixture used as an active ingredient in the present invention is a low-condensate of lactic acid derived from biological components, it has high biocompatibility and few side effects. The agent for suppressing or improving liver fibrosis according to the present invention is useful as a therapeutic agent for chronic liver disease.

Hereinafter, the implementation method and embodiment of this invention are demonstrated in detail.
The agent for suppressing or improving liver fibrosis according to the present invention includes a cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20 as an active ingredient, and includes, for example, liver fibrosis such as chronic liver disease. It can be used for the treatment or prevention of liver damage.

  The agent for suppressing or improving liver fibrosis according to the present invention is effective for suppressing liver fibrosis in chronic liver disorders such as viral hepatitis or alcoholic liver disorder. That is, the agent for suppressing or improving liver fibrosis of the present invention acts as a preventive agent for preventing liver fibrosis, a therapeutic agent for inhibiting the progression of liver fibrosis, and / or normalizing fibrosis. It has an effect as a therapeutic agent to recover.

  Specific examples of “liver damage” in the present specification include damage or destruction of liver tissue or cells, liver damage due to inflammation, decreased liver function, and liver failure. Specific examples of liver diseases associated with liver damage include, for example, alcoholic liver damage, cirrhosis, autoimmune hepatitis, congenital metabolic disorder liver damage, drug-induced hepatitis, biliary atresia, hepatitis A, hepatitis B, Examples include, but are not limited to, hepatitis C, hepatitis D, hepatitis E, hepatocellular carcinoma, primary biliary cirrhosis, and pyogenic liver abscess. The “liver disorder” referred to in the present invention is preferably a liver disorder accompanied by liver fibrosis. Specific examples of such liver disorder include viral hepatitis (hepatitis A, hepatitis B, hepatitis C, hepatitis D, Hepatitis, hepatitis E, etc.) or alcoholic liver disorder.

In the liver fibrosis inhibiting or improving agent of the present invention, a cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20 is used as an active ingredient.
The term “polylactic acid mixture” as used herein means a mixture in which cyclic and / or chain polylactic acid having a condensation degree of 3 to 20 is present in an arbitrary ratio. That is, the term “mixture” means a mixture of polylactic acids having any degree of condensation of 3 to 20, and at the same time, is used as a concept including a mixture of cyclic and chain polylactic acids. Such a “polylactic acid mixture” can be obtained by dehydrating and condensing lactic acid and purifying it by an appropriate method, as described hereinbelow. In the present specification, the term “polylactic acid mixture” is used for the sake of convenience. Among these, a single component such as cyclic polylactic acid having a certain degree of condensation or chain polylactic acid having a certain degree of condensation is used. Also included is polylactic acid.

  The degree of condensation means the number of lactic acid units that are repeating units in polylactic acid. For example, cyclic polylactic acid is presumed to have the following structural formula, where n represents the degree of condensation (ie, n = 3 to 20).

  When simply referred to herein as “lactic acid”, this lactic acid includes all of L-lactic acid, D-lactic acid or mixtures of these in any proportion. Preferably in the present invention, the lactic acid consists essentially of L-lactic acid. Here, “substantially” means that the ratio of L-lactic acid units in the polylactic acid mixture [ie, (L-lactic acid unit number / L-lactic acid unit number + D-lactic acid unit number) × 100] is 70, for example. % Or more, preferably 80% or more, more preferably 85% or more, still more preferably 90% or more, and particularly preferably 95% or more. The ratio of L-lactic acid units in the polylactic acid mixture depends on the ratio of L-lactic acid and D-lactic acid present in lactic acid used as a starting material.

  The method for producing a cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20 is not particularly limited. For example, JP-A-9-227388, JP-A-10-130153, It can be obtained by the production method described in Japanese Patent Application No. 11-39894 (all the contents described in these patent specifications are included as disclosure of the present specification by reference).

  More specifically, for example, a cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20 can be obtained by the following method A.

Method A:
First, lactic acid (preferably lactic acid substantially consisting of L-lactic acid) is subjected to dehydration condensation in an inert atmosphere. Examples of the inert atmosphere include nitrogen gas and argon gas, and it is preferable to use nitrogen gas.

  The dehydration condensation reaction is carried out under a reduced pressure of about normal pressure to about 1 mmHg and at a temperature of 110 to 210 ° C., preferably 130 to 190 ° C., but it is particularly preferable to carry out by a stepwise reduced pressure and a stepwise temperature increase. Although reaction time can be set suitably, for example, reaction can be performed for 1 to 20 hours. When stepwise pressure reduction and stepwise temperature rise are used, the reaction time is divided into partial reaction times consisting of two or more, and the reaction is carried out by setting the pressure and temperature in each part. When using stepwise pressure reduction, for example, normal pressure → 150 mmHg → 3 mmHg can be reduced, and when using stepwise temperature increase, for example, 145 ° C. → 155 ° C. → 185 ° C. can be raised. In practice, for example, a combination of these can be performed at 145 ° C. for 3 hours at atmospheric pressure, 145 ° C. for 3 hours at 150 mm Hg, 155 ° C. for 3 hours at 3 mm Hg, and 185 ° C. for 3 hours at 3 mm Hg for 1.5 hours it can.

  Subsequently, ethanol and methanol are added to the reaction mixture obtained by this dehydration condensation reaction, and the filtrate is dried by filtration to obtain an ethanol- and methanol-soluble component. That is, the term “ethanol and methanol soluble fraction” as used herein means a fraction soluble in a mixed solution of ethanol and methanol. In addition, when obtaining ethanol and methanol-soluble components, the reaction mixture of the dehydration condensation reaction is mixed with ethanol and methanol, and the ratio of ethanol and methanol at that time can be appropriately set. For example, ethanol: methanol = 1: 9. The order and method of adding ethanol and methanol to the reaction mixture are not limited and can be appropriately selected. For example, ethanol can be added first to the reaction mixture of the dehydration condensation reaction, and then methanol can be added. .

The ethanol / methanol-soluble matter obtained above is subjected to reverse phase column chromatography, particularly chromatography using an octadecylsilane (ODS) column, and first eluted with 25 to 50% by weight acetonitrile aqueous solution at pH 2 to 3. When the fraction is removed, and then the fraction eluted with 90% by weight or higher acetonitrile aqueous solution of pH 2-3, preferably 99% by weight or higher acetonitrile aqueous solution is collected, cyclic and / or chain having a condensation degree of 3 to 20 A polylactic acid mixture is obtained.
The cyclic and / or chain polylactic acid mixture obtained as described above is neutralized with an alkali substance such as sodium hydroxide, dried under reduced pressure, and then formulated into a desired form as described below by a conventional method. can do.

  As another method for producing a cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20 used in the present invention, for example, the method described in Japanese Patent Application No. 11-265715 (Method B and Or the method described in Japanese Patent Application No. 11-265732 (referred to as Method C) (the contents described in these patent specifications are all incorporated herein by reference). ). Hereinafter, the method B and the method C will be specifically described.

Method B:
In this method, a lactic acid oligomer is polymerized by polymerizing lactide in the presence of a lithium compound represented by RYLi (wherein R represents an aliphatic group or an aromatic group, and Y represents an oxygen atom or a sulfur atom). It is a manufacturing method. When performing a polymerization reaction, the usage-amount of a lithium compound (RYLi) is 1-0.1 mol per mol of lactide, Preferably it is a ratio of 0.2-0.3 mol. The reaction temperature is -100 to 0 ° C, preferably -78 to -50 ° C. The reaction is preferably started at a temperature of −78 to −50 ° C. and gradually raised to room temperature. The reaction is preferably carried out in the presence of a reaction solvent. As the reaction solvent, in addition to cyclic ethers such as tetrahydrofuran, diethyl ether, dimethoxyethane and the like can be used. As the reaction atmosphere, an inert gas atmosphere such as nitrogen gas or argon is used. The reaction pressure is not particularly limited and is preferably normal pressure.

  Note that the composition of the lactic acid oligomer obtained as described above (that is, the mixing ratio of the cyclic lactic acid oligomer and the chain lactic acid oligomer) varies depending on the lithium compound used as a reaction aid.

Method C:
In this method, (i) lactic acid is heated to a temperature in the range of 120 to 140 ° C. under a pressure condition of 350 to 400 mmHg to cause a dehydration condensation reaction, and only by-product water is distilled off without distilling lactide. A first heating step,
(Ii) After completion of the first heating step, the reaction product is heated to a temperature of 150 to 160 ° C., and the reaction pressure is reduced to 15 to 20 mmHg at a pressure reduction rate of 0.5 to 1 mmHg / min. Then, only by-product water was distilled off while avoiding the distillation of lactide, and after the reaction pressure dropped to 15-20 mmHg, the reaction was further continued under the same pressure condition and reaction temperature of 150-160 ° C. A second heating step for producing a dehydration condensate mainly comprising an oligomer;
(Iii) a third heating step in which, after completion of the second heating step, the chain lactic acid oligomer is cyclized by heating at 150 to 160 ° C. under a pressure condition of 0.1 to 3 mmHg to generate a cyclic oligomer;
It is the method characterized by comprising.

  In this method, first, in the first heating step, lactic acid is heated under reduced pressure to cause a dehydration condensation reaction. The reaction time in this case is 3 to 12 hours, preferably 5 to 6 hours. In the reaction under the first heating, by-product water generated by dehydration condensation of lactic acid is distilled off in order to make the reaction proceed smoothly. In this case, lactide which is a dehydration condensate of two molecules of lactic acid is removed. Carry out not to distill off. For this purpose, the reaction pressure is kept at a reduced pressure, preferably 300 to 500 mmHg, more preferably 350 to 400 mmHg, and heating is performed in the range of 100 to 140 ° C., preferably 130 to 140 ° C. under this pressure condition. Good. By the reaction in the first heating step, a reaction product mainly containing a dehydration condensate of 3 to 23 molecules of lactic acid is generated.

  After the completion of the first heating step, a temperature that is higher than the reaction temperature in the first heating step, preferably 145 to 180 ° C., so that an oligomer having an increased average polymerization degree is obtained in the second heating step. More preferably, while heating to a temperature of 150 to 160 ° C., the reaction pressure is lowered to a pressure of 10 to 50 mmHg, preferably 15 to 20 mmHg, and the dehydration condensation reaction is continued.

  As in the case of the reaction in the first heating step, this reaction is also carried out under the condition that by-product water is distilled off in order to make the reaction proceed smoothly, but lactide is not distilled off. The rate at which the reaction pressure is lowered to the pressure in the above range (pressure reduction rate) is 0.25 to 5 mmHg / min, preferably 0.5 to 1 mmHg in order to avoid the distillation of lactide and increase the reaction efficiency. It is usually necessary to keep in the range of / min. A pressure reduction rate lower than the above range is not preferable because it takes a long time to reduce the pressure to the predetermined pressure. On the other hand, a pressure reduction rate higher than the above range is not preferable because lactide distills together with by-product water. Absent.

After the reaction pressure has dropped to a predetermined pressure, the reaction is further continued at this reaction pressure. The heating time in this case is 3 to 12 hours, preferably 5 to 6 hours.
The reaction in the second heating step yields a lactic acid oligomer having an average degree of polymerization of 3 to 30, preferably 3 to 23. In this case, the ratio of the cyclic oligomer in the oligomer is usually 70 to 80% by weight. Degree.

After completion of the second heating step, in the third heating step, the reaction pressure is maintained at 0.25 to 5 mmHg, preferably 0.5 to 1 mmHg, and further reacted at a temperature of 145 to 180 ° C, preferably 150 to 160 ° C. Continue. The reaction time is 3 to 12 hours, preferably 5 to 6 hours. By-product water generated in this case is also distilled off. In this case, it is preferable to avoid evaporation of lactide, but since the reaction product contains almost no lactide, it is not necessary to reduce the rate of pressure reduction.
The reaction in the third heating step produces a lactic acid oligomer having an average degree of polymerization of 3 to 30, preferably 3 to 23, and a cyclic oligomer ratio of 90% by weight or more, preferably 99% by weight or more.

Method D:
The lactic acid mixture used in the present invention can also be produced by reacting lactide in the presence of an alkali metal compound represented by the formula (3): Me-N (R ¹ ) (R ² ). Hereinafter, Formula (3): Me-N (R ¹ ) (R ² ) will be described.

In the formula (3), Me represents an alkali metal. R ¹ and R ² each independently represents an aliphatic group or an aromatic group.
Examples of the aliphatic group include a saturated or unsaturated aliphatic hydrocarbon group having 1 to 12 carbon atoms, preferably 1 to 6 linear, branched, cyclic, or combinations thereof. And alkyl groups such as methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, octyl and dodecyl, and cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclooctyl and cyclododecyl. Is mentioned. The aliphatic group may be an unsaturated hydrocarbon group having a double bond or a triple bond.

  Here, examples of the aromatic group include aryl groups and arylalkyl groups having 6 to 30, preferably 6 to 20, more preferably 6 to 12, and further preferably 6 to 10 carbon atoms. Examples of the aryl group include phenyl, tolyl, naphthyl, and the like. Examples of the arylalkyl group include benzyl, phenethyl, naphthylmethyl, and the like.

  The aliphatic group and the aromatic group may have one or more substituents. The type of the substituent is not particularly limited, and examples thereof include a linear or branched, chain or cyclic alkyl group, a linear or branched, chain or cyclic alkenyl group, a linear or branched, chain or cyclic alkynyl group. , Aryl group, acyloxy group, alkoxycarbonyloxy group, aryloxycarbonyloxy group, carbamoyloxy group, carbonamido group, sulfonamide group, carbamoyl group, sulfamoyl group, alkoxy group, aryloxy group, aryloxycarbonyl group, alkoxycarbonyl Group, N-acylsulfamoyl group, N-sulfamoylcarbamoyl group, alkylsulfonyl group, arylsulfonyl group, alkoxycarbonylamino group, aryloxycarbonylamino group, amino group, ammonio group, cyano group, nitro group, carbo group A sil group, a hydroxyl group, a sulfo group, a mercapto group, an alkylsulfinyl group, an arylsulfinyl group, an alkylthio group, an arylthio group, a ureido group, a heterocyclic group (for example, containing at least one nitrogen, oxygen, sulfur, etc. 12-membered monocyclic ring, condensed ring), heterocyclic oxy group, heterocyclic thio group, acyl group, sulfamoylamino group, silyl group, halogen atom and the like. In the above, the alkyl, alkenyl, alkynyl and alkoxy generally have 1 to 12 carbon atoms, preferably 1 to 6 carbon atoms, and the aryl generally has 6 to 20 carbon atoms, preferably 6 to 6 carbon atoms. 10.

In the formula (3), Me represents an alkali metal. Examples of the alkali metal include Li, Na, and K, and Li is preferable.
The compound represented by formula (3) having an asymmetric carbon may be any of (R) isomer, (S) isomer, (R), (S) isomer.

  The method for obtaining the alkali metal compound represented by the formula (3) is not particularly limited, and those skilled in the art can appropriately obtain it. It can be obtained by reacting a dialkylamine such as diisopropylamine with an alkylated alkali metal such as n-butyllithium. More specifically, this reaction is carried out, for example, under a condition inert to the reaction such as under a nitrogen atmosphere, and with a solution containing a dialkylamine in an inert solvent such as THF and an alkyl in an inert solvent such as hexane. It can carry out by mixing and stirring with the solution containing alkali metal halide. The reaction temperature is not particularly limited as long as the reaction proceeds, but is preferably −78 ° C. to room temperature. The reaction time can be set as appropriate.

When the lactide is polymerized in the presence of the compound of the formula (3), the amount of the compound of the formula (3) (Me-N (R ¹ ) (R ² )) is preferably 1 to 0.00 per mole of lactide. 1 mol, more preferably 0.2 to 0.3 mol.

  Although the reaction temperature at the time of performing the polymerization reaction of lactide is not particularly limited as long as the reaction proceeds, it is preferably −100 ° C. to room temperature, more preferably −78 ° C. to room temperature.

  The polymerization reaction of lactide is preferably carried out in the presence of a reaction solvent. The reaction solvent is not particularly limited as long as it is inert to the reaction, but cyclic ethers such as tetrahydrofuran, diethyl ether, dimethoxyethane and the like can be preferably used. As the reaction atmosphere, an inert gas atmosphere such as nitrogen gas or argon gas can be used. The reaction pressure is not particularly limited and is preferably normal pressure.

  The composition of the mixture of the chain and cyclic lactic acid oligomers obtained by the above method varies depending on the kind of the compound of the formula (3) used as a reaction aid, reaction conditions, etc., but preferably the chain lactic acid rather than the cyclic lactic acid oligomer. High oligomer content.

  According to the method described above, the following formula (1) or (2):

(In the formula, m represents an integer of 1 to 18, and n represents an integer of 1 to 18)
A chain and cyclic lactic acid oligomer mixture represented by

  The above-mentioned methods A, B, C and D are only a part of specific examples of the method for producing the polylactic acid mixture used in the present invention, and in the present invention, the polylactic acid mixture produced by other methods. Can also be used.

  The agent for suppressing or improving liver fibrosis according to the present invention is used in preparations for pharmaceuticals, quasi-drugs and the like within the range not impairing the effects of the present invention, if necessary, in addition to the above essential components. Components and additives can be arbitrarily selected and used in combination. The agent for suppressing or improving liver fibrosis according to the present invention can be used in combination with pharmaceuticals and quasi-drugs in addition to being used as a single pharmaceutical.

The form of the agent for suppressing or improving liver fibrosis according to the present invention is not particularly limited, and it is possible to select an appropriate form most suitable for the purpose from the preparation forms for oral administration or parenteral administration. .
Examples of the dosage form suitable for oral administration include tablets, capsules, powders, drinks, granules, fine granules, syrups, solutions, emulsions, suspensions, chewables, etc. Examples of the dosage form suitable for parenteral administration include, but are not limited to, injections (subcutaneous injection, intramuscular injection, intravenous injection, etc.), external preparations, drops, inhalants, sprays, and the like. Never happen.

  Liquid preparations suitable for oral administration, for example, solutions, emulsions, syrups, etc., water, sugars such as sucrose, sorbit, fructose, glycols such as polyethylene glycol, propylene glycol, sesame oil, olive oil, soybean oil, etc. Oils, preservatives such as p-hydroxybenzoic acid esters, and flavors such as strawberry flavor and peppermint. For the production of solid preparations such as capsules, tablets, powders or granules, excipients such as lactose, glucose, sucrose and mannitol, starch, disintegrants such as sodium alginate, magnesium stearate, talc, etc. Lubricants, binders such as polyvinyl alcohol, hydroxypropyl cellulose, gelatin, surfactants such as fatty acid esters, plasticizers such as glycerin, and the like can be used.

  An injectable or infusion preparation suitable for parenteral administration preferably contains the above-mentioned substances as active ingredients dissolved or suspended in a sterile aqueous medium isotonic with the blood of the recipient. For example, in the case of an injection, a solution can be prepared using a salt solution, a glucose solution, or an aqueous medium composed of a mixture of salt water and a glucose solution. Formulations for enteral administration can be prepared using carriers such as cocoa butter, hydrogenated fat, or hydrogenated carboxylic acid and provided as suppositories. In the production of a spray, a carrier that can disperse the above-mentioned substance as an active ingredient as fine particles, does not irritate the recipient's oral cavity and airway mucosa, and facilitates the absorption of the active ingredient. Can be used. Specific examples of the carrier include lactose and glycerin. Depending on the substance that is the active ingredient and the nature of the carrier used, a formulation in the form of an aerosol or dry powder can be prepared. These preparations for parenteral administration include one or more selected from glycols, oils, flavors, preservatives, excipients, disintegrants, lubricants, binders, surfactants, plasticizers, and the like. Two or more kinds of foods and drinks can also be added.

The agent for suppressing or improving liver fibrosis according to the present invention can be applied to topical therapy such as intraarterial hepatic injection, embolization therapy, and the like in the form of a liquid or a fine particle dispersion. This agent can be used in combination with an appropriate effective trace metal such as zinc or copper.
The dose and frequency of administration of the agent for suppressing or improving liver fibrosis according to the present invention can be appropriately set according to various factors including the purpose of administration, dosage form, age, weight, sex, etc. of the intaker. However, the dose of the active ingredient is generally 1 to 10,000 mg / kg, preferably 10 to 2000 mg / kg, more preferably 10 to 200 mg / kg per day. It is preferable to administer the above-mentioned dosage in about 1 to 4 times a day.

The present invention further relates to a food or drink for suppressing liver fibrosis, which contains a cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20. That is, the cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20 used in the present invention is used not only in the form of a single preparation as described above but also in a food or drink. Can do.
If the food / beverage products of this invention can be mix | blended without decomposing | disassembling a polylactic acid mixture, there will be no restriction | limiting in particular in the compounding form.

  Specific examples of food and drink products of the present invention include soft drinks, drinks, health foods, foods for specified health use, functional foods, functionally active foods, dietary supplements, supplements, feeds, feed additives, etc. Mentioned health foods or supplements, including beverages.

Specific examples of food and drink include, for example, chewing gum, chocolate, candy, tablet confectionery, jelly, cookies, biscuits, yogurt and other confectionery, ice cream, ice confectionery and other frozen confectionery, tea, soft drinks (juice, coffee, cocoa Etc.), beverages such as nutritional drinks and beauty drinks, bread, ham, soup, jam, spaghetti, frozen foods and the like. Alternatively, the polylactic acid mixture used in the present invention can be added to seasonings or food additives. By ingesting the food / beverage products of the present invention, it is possible to provide a safe food / beverage product that exhibits an effect of suppressing liver fibrosis and exhibits substantially no harmful side effects.
The food / beverage products of the present invention can be obtained by directly mixing and dispersing a polylactic acid mixture in a general raw material used in foods and then processing it into a desired form by a known method.

  The food / beverage products of this invention include food / beverage products of all forms, The kind in particular is not restrict | limited, Various food / beverage products as described above, or various nutritional compositions, for example, various oral or enteral nutrition, A beverage or the like can be provided as a food or drink by blending the agent for suppressing or improving liver fibrosis according to the present invention. The composition of such foods and drinks can include proteins, lipids, carbohydrates, vitamins and / or minerals in addition to cyclic and / or chain polylactic acid mixtures having a condensation degree of 3 to 20. The form of the food or drink is not particularly limited, and may be any of solid, powder, liquid, gel, slurry and the like as long as it is easy to ingest.

Although content of the polylactic acid mixture in food-drinks is not specifically limited, Generally it is 0.1-20 weight%, More preferably, it is about 0.1-10 weight%.
The amount of the polylactic acid mixture contained in the food / beverage product is preferably included to such an extent that the liver fibrosis inhibitory effect of the present invention can be exerted, and preferably 0.1 g to 10 g in one food / drink taken. The degree is more preferably about 0.5 to 3 g.
The present invention will be described more specifically with reference to the following examples, but the present invention is not limited in any way by the examples.

Synthesis Example 1: Production of polylactic acid mixture (hereinafter also referred to as X01) 500 ml of L-lactic acid (which also contains D-lactic acid) was placed in a separable flask contained in a mantle heater. Nitrogen gas was introduced and stirred at 300 ml / min, and the distilled water was heated at 145 ° C. for 3 hours while being led to a flask with a reflux condenser through a temperature-reducing descending connecting tube. The pressure was further reduced to 150 mmHg and heated at the same temperature for 3 hours, and then heated at 155 ° C. for 3 hours under a reduced pressure of 3 mmHg and finally heated at 185 ° C. for 3 hours under reduced pressure of 3 mmHg to obtain polylactic acid as a reaction product It was.

  The obtained polylactic acid was kept at 100 ° C., and 100 ml of ethanol was added followed by 400 ml of methanol, followed by cooling. This was added to 500 ml of methanol, allowed to stand with sufficient stirring, and then purified by filtration. The filtrate was dried under reduced pressure and dissolved in acetonitrile to obtain a polylactic acid mixture. The resulting mixture of polylactic acid is in a state where a cyclic condensate and a linear condensate are mixed.

Synthesis Example 2: Production of polylactic acid mixture (hereinafter also referred to as X02) 10.0 g of (s)-(+)-lactic acid is placed in an eggplant-shaped flask having an internal volume of 100 ml, and this is set in a rotary evaporator. The pressure in the flask is adjusted to 350 to 400 mmHg, heated to 140 ° C., and the reaction is continued at the same pressure and the same temperature for 6 hours (first heating step). By-product water produced by this reaction was distilled off. Moreover, under the reaction conditions, lactide hardly distilled out of the system.

  Next, the reaction temperature was raised to 150 to 160 ° C., and the reaction pressure was gradually lowered from 400 mmHg over about 6 hours and dropped to 15 to 20 mmHg (pressure reduction rate: 1 mmHg / min). Under this pressure reduction rate, byproduct water was distilled off, but lactide was hardly distilled off. Thereafter, the pressure was maintained at 15 to 20 mmHg, and the reaction was continued for 6 hours (second heating step).

Next, the pressure was reduced to 1 to 3 mmHg over 30 minutes, and the reaction was continued for 5 hours at a reaction temperature of 160 ° C. (third heating step).
As a result of analyzing the reaction product after completion of the reaction, 6.80 g (yield 85%) of cyclic and chain oligomers having an average degree of polymerization of 3 to 20 were obtained.

Test example 1:
As test animals, Wister SPF rats (male, 5 weeks old) were used. First, rats were raised for 1 week in a feed (powder) containing 0.06% of 3′-methyl-4-dimethylaminoazobenzene (hereinafter abbreviated as DAB). Subsequently, the diet was returned to a normal diet, and a CCl ₄ + olive oil mixture (0.2 mL / 100 g body weight) was subcutaneously injected twice a week for 9 weeks. CCl ₄ was then discontinued and 10 mg / ml DAB / 0.5% w / v CMC-Na was orally administered daily for one week (0.4 ml / kg). This was followed by a normal diet for 2 weeks. Thereafter, the group that continued administration of the normal diet (normal diet administration group: Group II) and the group that started administration of the polylactic acid mixture (X01) produced in Synthesis Example 1 (CPL administration group: Group VIII) The test was conducted separately. For comparison, normal rats (Group I) not administered with DAB and CCl4 were also bred using a normal diet.

Liver tissues were collected from normal rats (Group I), rats (liver fibrosis induced by CCl ₄ + DAB) and CPL-administered groups (Group VIII), respectively. After fixation, paraffin sections were prepared. The section used a rabbit anti-MMP-13 (collagenase-3) polyclonal antibody (Chemicon International, Inc., CA, USA) as a primary antibody. Hereinafter, VECTASTAIN Elite ABC Rabbit IgG Kit and DAB Standard Kit (Vector Laboratories, Inc.) ., CA, USA) was used for immunohistochemical staining. Hematoxylin was used for counterstaining.

The results are shown in FIGS.
In the liver tissue (FIG. 1) of the untreated control group (Group I), the MMP-13 positive cell region and the negative cell region were mixed together in a lump. Furthermore, the brown positive part was observed in the form of granules in the liver cytoplasm.
In the liver tissue of Group II (FIG. 2), almost no MMP-13 positive site was observed.
In the liver tissue of group VIII (FIG. 3), MMP-13 positive cells accounted for the majority of hepatocytes. Intracellular positive sites were observed in the form of granules in the hepatocytes, similar to the findings in Group I.

  In rats, MMP-13 corresponds to MMP-1 in human liver tissue, and it is considered that the balance with TIMP-1 (Tissue Inhibitor of Metalloproteinase-1) is a trigger for liver fibrosis (Iredale, JP Gut 2000; 46: 443-446). The above results indicate that these balances are restored by administration of the polylactic acid mixture, and that the polylactic acid mixture is effective in suppressing liver fibrosis.

Test example 2:
In the same manner as in Test Example 1, according to the method already reported, 3′-methyl-4-dimethylaminoazobenzene (hereinafter abbreviated as DAB) was orally administered to Wistar rats (male, 5 weeks old) for 7 days (20 mg / day). kg × 2 / day), CCl ₄ + olive oil mixture (0.2 mL / 100 g body weight) was subcutaneously injected twice a week for 9 weeks, and DAB continuous oral administration was performed for 14 days. Two weeks after discontinuation of drug administration, necropsy was performed. After the appearance of liver fibers was confirmed, various drugs were administered for 4 weeks. A histological examination was performed after drug administration.

  The degree of fibrosis was classified into 5 grades of 0 to normal liver, 4 for cirrhosis, and 1 to 3 in accordance with the new Inuyama classification. The results of examining changes in fiberization using this classification are shown in Table 1 below.

The contents of each group in Table 1 are shown below.
Control: After the creation of cirrhosis, group supplemented with normal diet only: Additional sub-combination cycle of DAB + CCl ₄ administration SNMC: Widely used as an anti-inflammatory, fibrosis-preventing, and carcinogenesis-preventing drug for chronic hepatitis Group ARB administered with strong neo-minophagency (glycyrrhizin preparation): group administered with angiotensin receptor antagonist (Propress), which is reported to have anti-fibrotic action: Shisuito: anti-fibrosis / anti-inflammation . Group X02, which has been reported as a carcinogenic preventive and is actually used for cirrhosis and administered Shisui-to: group administered with the polylactic acid mixture (X02) produced in Synthesis Example 2

  As can be seen from the results in Table 1, it was found that the X02 group was effective not only in the control group but also in the fibrosis even when compared with a drug already considered effective in fibrosis. . In addition, in the X02 group, ballooning and the like were less with respect to hepatocellular injury than the control group and other drug groups, and the effectiveness was shown.

FIG. 1 shows the results of immunohistochemical staining for normal rat liver MMP-13. FIG. 2 shows the results of immunohistochemical staining for MMP-13 in the liver of rats in which liver fibrosis was induced (administered with a normal diet). FIG. 3 shows the result of immunohistochemical staining for MMP-13 in the liver of rats (administered with CPL) in which hepatic fibrosis was induced.

Claims (4)

An agent for suppressing or improving liver fibrosis, comprising a cyclic and / or chain polylactic acid mixture having a condensation degree of 3 to 20. The agent for suppressing or improving liver fibrosis according to claim 1 or 2, wherein lactic acid, which is a repeating unit in polylactic acid, substantially comprises L-lactic acid. The agent for suppressing or improving liver fibrosis according to claim 1 or 2, which is used for the treatment or prevention of liver damage associated with liver fibrosis. Food-drinks containing the suppression or improvement agent of liver fibrosis in any one of Claim 1 to 3.