KR100563616B1 - ?-Glucan derivative, manufacture of the derivative, adhesion Barrier using ?-glucan and method thereof - Google Patents

Abstract

The present invention relates to a beta glucan derivative, a method for preparing the same, and an anti-adhesion agent using a beta glucan derivative, based on a pachyman structure, which is a polysaccharide obtained from a mushroom, based on substitution of its glucose residue with acetamide. The anti-adhesion agent can adjust the biodegradable period, has excellent biocompatibility, excellent anti-adhesion effect, and has an effect of enhancing immune upon absorption in vivo.

Beta glucan derivatives, pachyman, anti-adhesion agents

Description

Beta-glucan derivatives, preparation method thereof and anti-adhesion agent using beta-glucan derivatives, preparation method thereof {β-Glucan derivative, manufacture of the derivative, adhesion Barrier using β-glucan and method

The present invention relates to a novel beta glucan derivative (AA-Pachman), a method for preparing the same, and an anti-adhesion agent using beta glucan derivatives through a water solubility control reaction of beta glucan, particularly, the cyclic polysaccharide pachyman. . Specifically, the present invention relates to a novel water-soluble polysaccharide obtained by binding an acetamide group to a glucose moiety of a polysaccharide that exhibits antitumor activity by increasing the immune function of the living body and increasing its stability and water solubility, and an economical method for producing the same from Fukryeong. In addition, the present invention relates to an anti-adhesion agent having excellent biocompatibility, excellent anti-adhesion effect, good biodegradability, and excellent immuno-enhancing effect when absorbed in vivo using the new polysaccharide.

Various kinds of polysaccharides are isolated from mushrooms and their antitumor activity is known and various studies have been made using them. Lentinan, a polysaccharide isolated from the fruiting body of shiitake mushrooms, Krestin, a protein polysaccharide isolated from the fruiting body and culture mycelium from shiitake mushrooms, and Shizophillan, a polysaccharide isolated from the culture filtrate It is commercially available as a drug showing antitumor activity and the like, and hydrothermal extracts of cultured mycelium of the situation are also produced and marketed in Korea.

Fukryeong is a fungal nucleus of Poria cocos belonging to the fungus of the Basidiomycetes, and its main ingredient is Pachyman, which was confirmed by Pachyman. It is insoluble in water and has difficulty in its use.

       Adhesion refers to a state in which tissues or organs of the human body are separated in a normal state but are not separated from each other by forming a fibrous tissue when a wound or an artificial force is applied, and a product that prevents such adhesion is an anti-adhesion agent.

       Surgical damage to tissues is naturally healed. At this time, the frequency of abnormal adhesions with surrounding tissues is 63-93% of surgical patients. Pain and infertility, as well as the difficulty of reopening surgery. For this reason, the development of anti-adhesion agents for preventing long-term adhesion after surgery is in progress worldwide.

  Commercially available anti-adhesion agents include Johnson & Johnson's Interceed® as a pad type, Adcon® by Gliatech, Seprafilm® by Genzyme, and SurgiWrap® by Macropore. Among them, Gliatech's Adcon® was discontinued in May 2002, and the technology was sold to Wright Medical Group, with very few anti-adhesion products on the market.

       Johnson & Johnson's Inerceed® is composed of oxidatively regenerated cellulose, which is composed of non-woven fabrics, and has a low solubility in water due to its high solubility in water. Genzyme's Seprafilm® is a chemically modified substance of carboxymethyl cellulose (CMC) and sodium hyaluronic acid, which can be used only in laparotomy and is functionally fragile, not highly transparent, and too sticky to tissues. There are disadvantages such as points. Macropore's SurgiWrap® is a polylactide-based component with high transparency and is widely used in recent years.It is functionally resistant to tissues and has a minimum duration of in vivo degradation of 6 months, which limits its use range. There are disadvantages such as being very expensive.

Hyaluronic acid (HA, USP 4,141,973) is an example of a biodegradable new material under development, and has a disadvantage in that it is decomposed and absorbed in vivo within a relatively fast time, and the price is also very expensive.

Technology based on carboxymethyl cellulose (CMC, carboxymethyl cellulose) or sodium carboxymethyl cellulose (SCMC) (KR publication 2003-55102, KR publication 2002-11955, KR publication 2001-10151, KR 10-316200, KR Patent No. 035707, WO 2001/05370, WO 2000/59516, WO 1998/13927) have been proposed, but this material does not show an anti-adhesion effect because it is dissolved and absorbed too quickly.

In situ gelation with sodium alginate (SAP 5,266,326) and hydroxypropyl methyl cellulose (HPMC, USP 5,318,780) has low adhesion and low complexity due to lack of tissue adhesion. There are disadvantages.

As a technique for overcoming the disadvantages of rapid degradation and absorption in vivo as described above, a method of forming crosslinks between biodegradable polymers has been proposed. The method using hyaluronic acid (HA) and carboxymethylcellulose (CMC) (USP 5,760,200) is used to react EDC [1-ethyl-3 (3-dimethyaminopropyl) carbo diimide hydrochloride] to maintain the hydrogel structure. The EDC is biotoxic, and the subsequent process to remove it is difficult, and the material HA is expensive, and the produced film is difficult to handle and procedure due to its low flexibility and strength.

Adhesion inhibitors based on carboxypolysaccharide (CPS, carboxypolysaccharide) and polyether (PE, polyether) (USP 5,906, 997, KR publication 2001-13927) have a disadvantage of poor adhesion to tissues after being saturated with water.

 Other gellan gums (KR publication 2003-55102, publication 2002-11955), chitosan (KR publication 2003-71119, publication 2001-107068), pullulan (KR publication 2003-71119), curdlan (curdlan, KR publication 2003-71119), glue (gellatin, KR publication 2001-107601), polyethylglycol (PEG, polyethylglycol, KR publication 2001-107067, KR publication 2001-107068), agarose (agarose, KR publication 2001-107067, published 2001-107068), dextran (KR special 035707), hydroxyethyl cellulose (HEC, hydroxyethyl cellulose, KR notification 1997-10545, notification 1995-13460), heparin, KR notification 1995- 13460) and the like using technologies such as materials are being developed.

       However, the above-mentioned materials have been attempted to be researched and developed alone or in combination of various materials due to their fast or slow degradability, but still have a limited function.

The present invention, in order to solve the conventional problems as described above, provides a novel polysaccharide with an anti-tumor activity in the mushroom and easy to use water-soluble controlled polysaccharide and a method for preparing the same, the biodegradable period of the conventional anti-adhesion agent It is an object of the present invention to solve the problem, to improve the anti-adhesion effect, and to provide an anti-adhesion agent of a new material which enhances immune function after decomposition and absorption.

The present invention to achieve the above object.

It provides a beta glucan derivative represented by the following formula (1) characterized in that its glucose residue is substituted with acetamide based on the pachyman structure, which is a polysaccharide obtained from a mushroom.

In Formula 1, R and R 'is an acetamide group (-CH ₂ CONH ₂ ), m and n are the number of each monomer, m is 40 to 60, n is in the range of 60 to 100.

The polysaccharide is a beta glucan derivative, characterized in that it comprises a bokyeong polysaccharide, low polysaccharide and yeast polysaccharide.

In addition, the present invention provides a beta glucan derivative, characterized in that the degree of substitution of the acetamide is 0.01 to 0.5.

In addition, the present invention (a) extracting the dry powder of the mushrooms with an alkali to separate the polysaccharide; (b) adding and stirring a reaction solvent of a lower alcohol containing isopropanol to the extracted polysaccharide of step (a); (c) selecting from an alkaline solution containing sodium hydroxide or calcium hydroxide in the solution of step (b) and adding 0.1 to 50% of the total weight as a neutralizing agent to stir; (d) To the solution of step (c) is selected from chloro compounds such as chloroacetic acid, chloroacetamide, dichloroacetamide, trichloroacetamide, chloroacetyl chloride, chloroacrylic acid and the like as a reaction material to 30 ~ 80 ℃ Reacting at a reaction temperature; (e) providing a method for producing a beta glucan derivative comprising the step of washing the precipitate obtained by centrifuging the solution of step (d) with an alcohol solvent and drying under reduced pressure.

In another aspect, the present invention provides an anti-adhesion agent comprising a beta glucan derivative represented by the following formula (1) in which a glucose residue is substituted with acetamide based on a pachyman structure, which is a polysaccharide obtained from a mushroom. .

Formula 1.

In addition, the present invention provides an anti-adhesion agent, characterized in that the beta glucan derivative contains 1 to 30% by weight of metal ions.

In addition, the present invention provides a beta glucan derivative, characterized in that the degree of substitution of the acetamide is 0.01 to 0.5.

In another aspect, the present invention provides an anti-adhesion agent comprising a beta glucan derivative having a powder, gel, thin film form.

In addition, the present invention comprises the steps of suspending a beta glucan derivative in water; homogenizing the suspension with a microfluidizer; The homogenized suspension is poured into a glass plate and dried in a clean bench to provide a method for producing a thin film anti-adhesion agent.

In addition, the present invention comprises the steps of suspending a beta glucan derivative having immunostimulating activity and water-soluble in water: homogenizing the suspension; It provides a method for producing a thin film type anti-adhesion agent comprising the step of preparing a thin film by the coating machine to the homogenized suspension.

The beta glucan derivative has immunostimulating activity, little cytotoxicity, and water solubility is controlled.

In the beta glucan derivative of the present invention, the degree of substitution of acetamide group is preferably 0.01-1.0, and most preferably, when the degree of substitution is 0.2-0.4. Physical and chemical properties of the beta glucan derivatives of the present invention are as follows.

(1) appearance

The beta glucan derivative of the present invention is white or pale yellow white, tasteless and odorless.

(2) molecular weight

The molecular weight range of the beta glucan derivative of the present invention is 5-20 x 10 ⁶ and the average molecular weight is 8.7 x 10 ⁶ .

(3) melting point

The beta glucan derivative of the present invention is characterized by being carbonized at a high temperature of 170 ° C. or more without melting point.

(4) solubility

The beta glucan derivatives of the present invention have very low solubility in water and form gels when contained in water, and do not dissolve in methanol, ethanol, butanol, acetone, ethyl acetate, methylene chloride, chloroform, hexane, ether and petroleum ether. .

(5) infrared absorption spectrum

Beta-glucan derivative of the present invention is analyzed by infrared absorption spectrum using potassium bromide disk method wave number 3,400cm ^-1 , 2,920cm ^-1 , 1,650cm ^-1 , 1,600cm ^-1 , 1,470-1,400cm ^-1 , 1,100-1,000 cm ^-1, the absorption band appears at 890cm ^-1. At this time, the absorption band of 890 cm ^-1 is due to beta-glycoside.

(6) total sugar analysis

The total sugars of the beta glucan derivatives of the present invention are analyzed by the phenol-sulfuric acid method, and the analysis shows that 100% are all sugars.

(7) total protein analysis

The total protein of the beta glucan derivative of the present invention is analyzed by Lowry-Folin method, and the analysis result is 0%.

The present invention also provides a method for producing a beta glucan derivative exhibiting an anti-adhesion effect. Specifically, the present invention extracts the dry powder of Bokryeong with alkali according to a method such as Saito to separate the Bokryeong polysaccharide. The beta glucan derivative is prepared by adding a reaction solvent to the Bokyong polysaccharide, stirring the mixture, adding a neutralizing agent, stirring the mixture, and reacting by adding the reactant. The precipitate obtained by centrifugation is washed with a suitable solvent and dried under reduced pressure.

  Hereinafter, the present invention will be described in detail by way of examples. The following examples are illustrative of the invention and are not intended to limit the scope of the invention.

Example 1 Preparation of Beta Glucan Derivative

Comparative Example 1 Acetamino Group Substitution Degree 0.5

       1 g of beta glucan obtained from Bokryeong was placed in a reactor, 30 ml of isopropanol was added thereto, shaken at room temperature for 30 minutes, and 4.6 ml of 10% sodium hydroxide solution was added at a rate of 1 ml in 10 minutes. At this time, the mixture was vigorously shaken for 90 minutes so that no gel was formed, and 0.375 g of chloroacetamide was added thereto and reacted at 50-60 ° C. for 1 hour. After the reaction, the reaction solution was washed with 20 ml of 70% ethanol added with 10 ml of acetic acid and again washed with 80% ethanol and ethanol anhydrous in sequence. The precipitated polysaccharide was separated and dried under reduced pressure to obtain 0.85 g of white glucan derivative. The derivative had a solubility in water of at least 5%, and no precipitate was formed even when stored for 3 months in a 5% aqueous solution.

       In order to investigate the anti-adhesion effect of the beta glucan derivative prepared in this comparative example, a thin film was prepared, which was used as the first sample.

Comparative Example 2 Acetamino Group Substitution Degree 0.4

       1 g of beta glucan obtained from Bokryeong was placed in a reactor, 30 ml of isopropanol was added thereto, shaken at room temperature for 30 minutes, and 3.68 ml of 10% sodium hydroxide solution was added at a rate of 1 ml in 10 minutes. At this time, vigorously shaken for 90 minutes so that no gel was formed, and 0.3 g of chloroacetamide was added thereto and reacted at 50-60 ° C. for 1 hour. After the reaction, 10 ml of acetic acid was added and washed with 200 ml of 70% ethanol, and then washed again with 80% ethanol and ethanol anhydrous. Precipitated polysaccharide was separated and dried under reduced pressure to obtain 0.82 g of white glucan derivative. The derivative had a solubility in water of 0.5%, and no precipitate was formed even when stored for 3 months in a 0.5% aqueous solution.

       In order to investigate the anti-adhesion effect of the beta glucan derivative prepared in this comparative example, a thin film was prepared, which was used as the second sample.

Comparative Example 3 Acetamino Group Substitution Degree 0.3

       1 g of beta glucan obtained from Bokryeong was placed in a reactor, 30 ml of isopropanol was added thereto, shaken at room temperature for 30 minutes, and 2.76 ml of 10% sodium hydroxide solution was added at a rate of 1 ml in 10 minutes. At this time, the mixture was vigorously shaken for 90 minutes to prevent gel formation, and 0.225 g of chloroacetamide was added thereto and reacted at 50-60 ° C. for 1 hour. After the reaction, 10 ml of acetic acid was added and washed with 200 ml of 70% ethanol, and then washed again with 80% ethanol and ethanol anhydrous. The precipitated polysaccharide was separated and dried under reduced pressure to obtain a 0.83 g white glucan derivative. The derivative did not dissolve in water and formed a gel.

       In order to investigate the anti-adhesion effect of the beta glucan derivative prepared in this comparative example, a thin film was prepared, which was used as sample 3.

Comparative Example 4 Acetamino Group Substitution Degree 0.2

       1 g of beta glucan obtained from Bokryeong was placed in a reactor, 30 ml of isopropanol was added thereto, shaken at room temperature for 30 minutes, and 9.2 ml of 2% sodium hydroxide solution was added at a rate of 1 ml in 10 minutes. At this time, the mixture was vigorously shaken for 90 minutes to prevent gel formation, and 0.15 g of chloroacetamide was added thereto and reacted at 50-60 ° C. for 1 hour. After the reaction, 10 ml of acetic acid was added and washed with 200 ml of 70% ethanol, and then washed again with 80% ethanol and ethanol anhydrous. The precipitated polysaccharide was separated and dried under reduced pressure to obtain a beta glucan derivative, 0.7 g of white powder. The derivative did not dissolve in water and did not form a gel.

       In order to investigate the anti-adhesion effect of the beta glucan derivative prepared in this comparative example, a thin film was prepared, which was used as the fourth sample.

Comparative Example 5 Acetamino Group Substitution Degree 0.1

       1 g of beta glucan obtained from Bokryeong was placed in a reactor, 30 ml of isopropanol was added thereto, and shaken at room temperature for 30 minutes. Then, 4.6 ml of 2% sodium hydroxide solution was added at a rate of 1 ml in 10 minutes. At this time, the mixture was vigorously shaken for 90 minutes so that no gel was formed, and 0.075 g of chloroacetamide was added thereto and reacted at 50-60 ° C. for 1 hour. After the reaction, 10 ml of acetic acid was added and washed with 200 ml of 70% ethanol, and then washed again with 80% ethanol and ethanol anhydrous. The precipitated polysaccharide was separated and dried under reduced pressure to obtain a beta glucan derivative, 0.7 g of white powder. The derivative did not dissolve in water and did not form a gel.

       In order to investigate the anti-adhesion effect of the beta glucan derivatives prepared in this comparative example, a thin film was prepared, which was used as the fifth sample.

Comparative Example 6 Acetamino Group Substitution Degree 0.06

       1 g of beta glucan obtained from Bokryeong was placed in a reactor, 30 ml of isopropanol was added thereto, shaken at room temperature for 30 minutes, and 2.8 ml of 2% sodium hydroxide solution was added at a rate of 1 ml in 10 minutes. At this time, the mixture was vigorously shaken for 90 minutes to prevent gel formation, and 0.045 g of chloroacetamide was added thereto and reacted at 50-60 ° C. for 1 hour. After the reaction, 10 ml of acetic acid was added and washed with 200 ml of 70% ethanol, and then washed again with 80% ethanol and ethanol anhydrous. The precipitated polysaccharide was separated and dried under reduced pressure to obtain 0.74 g of a white powder of beta glucan derivative. The derivative did not dissolve in water and did not form a gel.

       In order to investigate the anti-adhesion effect of the beta glucan derivative prepared in this comparative example, a thin film was prepared, which was used as the sixth sample.

Comparative Example 7 Acetamino Group Substitution Degree 0.03

       1 g of beta glucan obtained from Bokryeong was placed in a reactor, 30 ml of isopropanol was added thereto, shaken at room temperature for 30 minutes, and 1.4 ml of 2% sodium hydroxide solution was added at a rate of 1 ml in 10 minutes. At this time, the mixture was vigorously shaken for 90 minutes so that no gel was formed, and 0.0225 g of chloroacetamide was added thereto and reacted at 50-60 ° C. for 1 hour. After the reaction, wash with 200 ml of 70% ethanol added with 10 ml of acetic acid, and again with 80% ethanol and ethanol anhydrous.

Washed out as usual. The precipitated polysaccharide was separated and dried under reduced pressure to obtain a beta glucan derivative of 0.76 g of white powder. The derivative did not dissolve in water and did not form a gel.

       In order to investigate the anti-adhesion effect of the beta glucan derivative prepared in this Comparative Example, a thin film was prepared, which was used as the seventh sample.

[Comparative Example 8] Beta-glucan of the Booryeong

       The beta glucan itself obtained from Bokryeong was used. It did not dissolve in water and did not form a gel.

       In order to compare the anti-adhesion effect with the beta glucan derivative prepared in this Comparative Example, a thin film was prepared, which was used as the eighth sample.

Example 2 Thin Film Preparation of Beta Glucan Derivative

       0.5 g of each derivative and beta glucan obtained in Comparative Examples 1 to 8 of Example 1 were suspended in 50 ml of water, homogenized using a microfluidizer, and then poured into a glass plate 18 cm wide and clean. A thin film was prepared by drying in a bench for 1 day to obtain samples 1-8.

Experimental Example 1 Safety of in vitro glucan derivatives ( in vitro , cytotoxicity)

       In order to investigate the cytotoxicity of each derivative obtained in Comparative Examples 1-8 of Example 1, 3- (4,5-dimethyldiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT MTT search method using). HS27 cells derived from human normal cells were adjusted to phosphate buffer solution (PBS) to a concentration of 1 × 10 5 cells / ml and placed in 100 μl per well in a 96-well microplate (Falcon, USA). Next, the derivative powders 1-8 were suspended in a phosphate buffer solution at a concentration of 1 mg / ml, diluted 1/10 to continue dilution until the concentration was 0.1 µg / ml, and the final volume was adjusted to 200 µl at 37 ° C., Incubated for 48 hours in a 5% CO2 incubator. Next, incubate for 4 hours by adding 20 μl of a phosphate buffer solution containing 5 mg / ml of MTT reagent, remove 180 μl of supernatant, and add 100 μl of 0.04N hydrochloric acid-isopropanol to stand overnight. Cell viability was measured by measuring the absorbance at 560 nm using an automatic eliza reader (Molecular Devices Corp., USA).

Mean survival rate (%) = absorbance of sample treated group / absorbance of control group x 100

As a result, the glucan derivative of the present invention did not show direct cytotoxicity and was found to be a safe substance (see Table 1).

Direct Cytotoxicity of the Glucan Derivatives of the Invention on HS27 Cell Line Sample Name Sample concentration (㎍ / ㎖) Cell average survival rate (%) Sample 1 1,000 102.5 100 101.7 10 107.7 One 100.2 0.1 101.5 Sample 2 1,000 110.2 100 102.3 10 100.8 One 100.2 0.1 108.9 Sample 3 1,000 105.4 100 103.7 10 100.2 One 100.9 0.1 111.1 Sample 4 1,000 108.2 100 100.7 10 105.3 One 104.7 0.1 107.3 Sample 5 1,000 104.2 100 100.9 10 108.1 One 106.4 0.1 107.7 Sample 6 1,000 100.1 100 102.2 10 108.5 One 104.3 0.1 102.7 Sample 7 1,000 108.4 100 109.2 10 100.8 One 102.9 0.1 106.5 Sample 8 1,000 104.7 100 107.9 10 108,2 One 100.3 0.1 105.4

Experimental Example 2

Safety of beta glucan derivatives ( in vivo , animal oral toxicity)

       Oral toxicity test was conducted to investigate the safety of each derivative obtained in Comparative Examples 1-8 of Example 1. Animals were used as rats (ICR mouse), and rats with an average body weight of 18 g were purchased for 1 week, and 10 rats were used in one group. In detail, eight derivatives were suspended in water, and the rats, which had been adapted for one week, were forcibly administered orally at a dose of 1,000 mg / kg for two weeks. It was. As a result, all 80 animals in 8 groups showed no symptoms.

Experimental Example 3

Anti-Adhesion Effect of Beta Glucan Derivative Film ( in vivo , Animal Model)

Sample 3-1 obtained in Comparative Example 3 of Example 1 and sample 3-1 added calcium ion (Ca ²⁺ ), which is a metal ion of 5% (W / W) weight of the derivative 3, were used. 230 g of Sprague-Dawley (SD) rat females were used for the test. SD rats were anesthetized with diethyl ether, and then a clean surgical instrument was used to open the center of the abdomen to expose the small intestine and scratch the surface to cause bleeding. The corresponding abdominal wall was rubbed with sandpaper to bleed. After causing spots, the control group of sample No. 3 adhered the thin film to the wound area, and then relocated the abdominal wall and skin layer adjacent to the abrasion site of the organ and the wound site of the abdominal wall and sutured the abdominal wall and skin layer by general surgery. . At this time, each treatment group was 10 rats, and after 7 days, the rats were euthanized and then opened to confirm the prevention of adhesion, shape retention, and abnormal change of organs.

       The degree of adhesion was determined using the following grading system. The criteria for adhesion was based on adhesion to the abdominal wall.

Class 0: No adhesion

Grade 1: Abdominal fat adheres slightly to the wound

Level 2: The mesentery and the wound are in contact with each other.

Grade 3: Vascular tissue is developed and the small intestine and abdominal wall are heavily adhered

       Thin film shape maintenance degree was based on the thin film shape.

Class 0: No change of form

Grade 1: A little gelation caused the thin film to be thick but retained form.

Level 2: Gelation does not maintain the shape of the thin film, but the gelled material remains intact

Grade 3: Gelation occurs and some substances are decomposed and absorbed.

       As a result, it showed a significant anti-adhesion effect compared to the average degree of adhesion of the control group, there was no abnormal change of organ. In particular, in the case of sample 3-1 with calcium, the shape retention rate was better than that in sample 3.

Anti-adhesion Effect, Thin Film Form Retention Rate Test of Glucan Derivative of the Present Invention sample Rating Adhesion prevention effect  Thin Film Form Retention Rate Number of individuals Average rating Number of individuals Average rating  Control 0 0  (1 × 1 + 2 × 2 + 3 × 7) / 10 = 2.6 -   - One One - 2 2 - 3 7 -  Sample 3 0 7  (1 × 2 + 2 × 1) / 10 = 0.4 8  (1 × 2) / 10 = 0.2 One 2 2 2 One 0 3 0 0  3-1 sample 0 8  (1 × 2) / 10 = 0.2 10  0.0 One 2 0 2 0 0 3 0 0

Beta-glucan derivatives according to the present invention exhibits anti-tumor activity by enhancing the immune activity of the living body, so there is almost no cytotoxicity. .

In addition, the beta glucan derivatives according to the present invention can be obtained through a simple manufacturing method, the production method is economical because it is low cost, the beta glucan derivatives prepared using chloroacetamide as a reactant is prepared Ease of use, excellent stability, there is an effect of enhancing the immune effect.

In addition, the anti-adhesion agent according to the present invention can adjust the period of biodegradability, has excellent biocompatibility, excellent effect of anti-adhesion, and has an immune enhancing effect upon absorption in vivo.

Claims (13)

A beta glucan derivative represented by the following formula (1) characterized in that its glucose residue is substituted with acetamide on the basis of the polysaccharide pachyman structure. Formula 1

In Formula 1, R and R 'is an acetamide group (-CH ₂ CONH ₂ ), m and n are the number of each monomer, m is 40 to 60, n is in the range of 60 to 100. [Claim 2] The beta glucan derivative according to claim 1, wherein the polysaccharide comprises a plural polysaccharide, an oligosaccharide, and a yeast polysaccharide. The beta glucan derivative according to claim 1, wherein the degree of substitution of acetamide is 0.01 to 0.5. (a) extracting the dry powder of the mushroom with alkali to separate the polysaccharide; (b) adding and stirring a reaction solvent of a lower alcohol including isopropanol to the polysaccharide extracted in step (a); (c) selecting from an alkaline solution containing sodium hydroxide or calcium hydroxide in the solution of step (b) and adding 0.1 to 50% of the total weight as a neutralizing agent to stir; (d) To the solution of step (c) is selected from chloro compounds such as chloroacetic acid, chloroacetamide, dichloroacetamide, trichloroacetamide, chloroacetyl chloride, chloroacrylic acid and the like as a reaction material to 30 ~ 80 ℃ Reacting at a reaction temperature; (E) a method for producing a beta glucan derivative of formula (I) comprising the step of washing the precipitate obtained by centrifuging the solution of step (d) with an alcohol solvent and drying under reduced pressure. In the anti-adhesion agent which prevents abnormal adhesion of organs, The anti-adhesion agent is an anti-adhesion agent using a beta glucan derivative, characterized in that it comprises a beta glucan derivative represented by the following formula (1) in which a glucose residue is substituted with acetamide based on a pachyman structure which is a polysaccharide obtained from a mushroom. . Formula 1

In Formula 1, R and R 'is an acetamide group (-CH ₂ CONH ₂ ), m and n are the number of each monomer, m is 40 to 60, n is in the range of 60 to 100. 6. The anti-adhesion agent using beta glucan derivatives according to claim 5, wherein the beta glucan derivative contains metal ions in a weight ratio of 0.1 to 30%. 6. The anti-adhesion agent using beta glucan derivatives according to claim 5, wherein the polysaccharide comprises a plural polysaccharide, an oligosaccharide, and a yeast polysaccharide. The anti-adhesion agent using beta glucan derivatives according to claim 5, wherein the degree of substitution of acetamide is 0.01 to 0.5. The anti-adhesion agent according to claim 5, wherein the anti-adhesion agent has a powder, a gel, or a thin film form. The method of claim 5, The beta glucan derivative is an anti-adhesion using a beta glucan derivative, characterized in that it has an immune enhancing activity. Suspending a beta glucan derivative in which a glucose residue is substituted with acetamide based on a pachyman structure in water: Homogenizing the suspension; And Method for preparing an anti-adhesion using a beta glucan derivative of formula 1 characterized in that it comprises the step of drying the homogenized suspension in a clean bench. Suspending a beta glucan derivative having immunopotentiating activity and water solubility in water: Homogenizing the suspension; and Method for producing an anti-adhesion agent using a beta glucan derivative of formula 1 characterized in that it comprises the step of preparing a thin film by the homogenized suspension coating machine. The method according to claim 11 or 12, wherein Homogenizing the suspension is a method of preparing an anti-adhesion agent using a beta glucan derivative of the formula (1), characterized in that using a microfluidizer (microfluidizer).