TW202434316A - Anti-sticking material - Google Patents

Abstract

The purpose of the present invention is to provide an anti-adhesion material that can exert an excellent anti-adhesion effect. The present invention is an anti-adhesion material, comprising a cross-linked gelatin gel having an uncross-linked lysine residue number of 1.15×10 ^-4 mol/g to 2.50×10 ^-4 mol/g.

Description

Anti-sticking material

The present invention relates to an anti-sticking material having an excellent anti-sticking effect.

Background Technology

Adhesion refers to a state in which continuity occurs between free organs or tissues that originally existed close to each other. Postoperative suture adhesion is a type of inflammatory adhesion that is artificially produced. Although the degree varies, it is still a complication that is likely to occur due to surgery. In particular, it is also known that if tendon adhesion occurs after treatment of tendon injury, tendon rupture, fracture, etc., the joint will become immobile and the movable area of the joint will be reduced. In order to restore the damaged tissue normally, it is very important to prevent tendon adhesion after surgery by using adhesion prevention materials.

Anti-adhesion materials are required to be able to be injected into the area where adhesion is to be prevented, and to stay in the area for a certain period of time to function as a physical barrier. In particular, when preventing tendon adhesion, since the surrounding tissue of the tendon is both fine and complex, it is required to have excellent diffusion properties during injection so that it can spread throughout the entire target area, and it is required to stay in the target area for a certain period of time after injection to function as a physical barrier.

To date, there have been proposals for anti-adhesion materials using various materials. For example, in Patent Document 1, it is described that a fragmented polymer composition containing a cross-linked hydrogel having a subunit size, balanced swelling, and in vivo decomposition time within a specified range can be used for the purpose of anti-adhesion, and a cross-linked gelatin hydrogel can be used as the cross-linked hydrogel. Known Technical Documents Patent Documents

[Patent Document 1] Japanese Patent Publication No. 2002-515086

Invention Summary Problem to be solved by the invention

The anti-adhesion materials using cross-linked gelatin gels reported so far have the disadvantages of not being able to fully prevent adhesion and not being able to improve the joint movable area when used for the purpose of preventing tendon adhesion. In recent years, the demand for improved functionality of anti-adhesion materials has increased, and the development of novel anti-adhesion materials has been pursued.

Therefore, the purpose of the present invention is to provide an anti-adhesion material that can exert an excellent anti-adhesion effect. Means for solving the problem

In cross-linked gelatin gels, cross-linking occurs via the ε-amine groups of lysine residues. Therefore, the number of uncross-linked lysine residues in cross-linked gelatin gels is an indicator of the degree of cross-linking.

After conducting a full-hearted study to solve the aforementioned problem, the inventors found that by using a cross-linked gelatin gel with a cross-linking degree controlled within a specific range, tissue adhesion can be effectively prevented. Furthermore, if used for the purpose of preventing tendon adhesion, the joint movable zone can be effectively improved. Specifically, it was found that a cross-linked gelatin gel with an uncross-linked lysine residue number of 1.15×10 ^-4 mol/g~2.50×10 ^-4 mol/g (i.e., a cross-linking degree of 30~67%) has an excellent adhesion prevention effect. Furthermore, if used for the purpose of preventing tendon adhesion, the joint movable zone can be effectively improved. In addition, the inventors have also learned that by using a cross-linked gelatin gel with a pepsin digestion time of 90 to 320 minutes measured by the specified method described below, tissue adhesion can be effectively prevented. Furthermore, if used for the purpose of preventing tendon adhesion, the joint movable area can be effectively improved. The present invention is based on these insights and further repeated studies.

That is, the present invention provides the invention of the following disclosed aspects. Item 1-1. An anti-adhesion material comprising a cross-linked gelatin gel having an uncross-linked lysine residue number of 1.15×10 ^-4 mol/g to 2.50×10 ^-4 mol/g. Item 1-2. An anti-adhesion material as described in Item 1-1, wherein the pepsin digestion time of the cross-linked gelatin gel determined by the following measurement method is 90 to 320 minutes. <Method for measuring pepsin digestion time> Prepare a dry powder of the cross-linked gelatin gel to be measured. Mix 0.02 g of the dried cross-linked gelatin powder and 0.08 g of a solvent (physiological saline solution to which polyethylene glycol is added in a manner to become 10 g/100 ml) to prepare a measurement sample. Separately, pepsin from pig gastric mucosa is added to 0.1N hydrochloric acid aqueous solution to dissolve it in a concentration of 1200 units/ml, and the solution is filtered to prepare a pepsin solution. 0.1 g of the sample is added to 40 ml of the pepsin solution, and the mixture is stirred at 37°C in an incubator using a stirrer while the time until the cross-linked gelatin (insoluble matter) disappears is determined as the pepsin digestion time. Item 1-3. The anti-adhesion material as described in Item 1-1 or 1-2, wherein the cross-linked gelatin gel is a hydrogel. Item 1-4. The anti-adhesion material as described in Item 1-3, wherein the hydrogel is a paste. Item 1-5. The anti-adhesion material as described in Item 1-1 or 1-2, wherein the cross-linked gelatin gel is an aerogel, and the anti-adhesion material is a two-agent type preparation, wherein the two-agent type preparation contains a first preparation comprising the cross-linked gelatin gel and a second preparation comprising an aqueous solvent. Item 1-6. The anti-adhesion material as described in any one of Items 1-1 to 1-5 is used for preventing adhesion of tendons. Item 1-7. An anti-adhesion method, comprising administering an effective amount of the anti-adhesion material described in any one of Items 1-1 to 1-6 to a site where adhesion of biological tissues needs to be prevented. Item 1-8. A cross-linked gelatin gel having an uncross-linked lysine residue number of 1.15×10 ^-4 mol/g to 2.50×10 ^-4 mol/g is used for the manufacture of an anti-adhesion material. Item 1-9. A cross-linked gelatin gel having an uncross-linked lysine residue number of 1.15×10 ^-4 mol/g to 2.50×10 ^-4 mol/g is used for a treatment for preventing adhesion of biological tissues.

In addition, the present invention provides the invention of the following disclosed aspects. Item 2-1. An anti-adhesion material comprising a cross-linked gelatin gel having a pepsin digestion time of 90 to 320 minutes as measured by the measurement method shown in the aforementioned Item 1-2. Item 2-2. The anti-adhesion material as described in Item 2-1, wherein the cross-linked gelatin gel is a hydrogel. Item 2-3. The anti-adhesion material as described in Item 2-2, wherein the hydrogel is a paste. Item 2-4. The anti-adhesion material as described in Item 2-3, wherein the cross-linked gelatin gel is an aerogel, and the anti-adhesion material is a two-agent type preparation, and the two-agent type preparation contains a first preparation containing the cross-linked gelatin gel and a second preparation containing an aqueous solvent. Item 2-5. The anti-adhesion material as described in any one of Items 2-1 to 2-4 is used for preventing adhesion of tendons. Item 2-6. An anti-adhesion method is to administer an effective amount of the anti-adhesion material described in any one of Items 2-1 to 2-5 to a site where adhesion of biological tissues needs to be prevented. Item 2-7. A cross-linked gelatin gel having a pepsin digestion time of 90 to 320 minutes as measured by the measurement method shown in Item 1-2 is used for the manufacture of an anti-adhesion material. Item 2-8. A cross-linked gelatin gel having a pepsin digestion time of 90 to 320 minutes as measured by the measurement method shown in Item 1-2 is used for a treatment for preventing adhesion of biological tissues. Effect of the invention

The anti-adhesion material of the present invention can exert a particularly excellent anti-adhesion effect by using a cross-linked gelatin gel with specific physical properties. In addition, the anti-adhesion material of the present invention can also effectively improve the movable area of the joint if used to prevent tendon adhesion of the joint.

The form used to implement the invention

1. First Implementation Form of Anti-Adhesion Material In one implementation form of the anti-adhesion material of the present invention, it is characterized by comprising a cross-linked gelatin gel having an uncross-linked lysine residue number of 1.15×10 ^-4 mol/g to 2.50×10 ^-4 mol/g. The anti-adhesion material of this implementation form is described in detail below.

[Crosslinked gelatin gel] ・Number of uncrosslinked lysine residues The number of uncrosslinked lysine residues of the crosslinked gelatin gel used in the present invention is 1.15×10 ^-4 mol/g to 2.50×10 ^-4 mol/g. In the present invention, "number of uncrosslinked lysine residues" refers to the molar number of lysine residues having uncrosslinked ε-amino groups per 1g of the dry weight of the crosslinked gelatin gel, and is a physical property value that serves as an index of the degree of crosslinking. In the adhesion prevention material of the present invention, by using a cross-linked gelatin gel having an uncross-linked lysine residue number within the above-mentioned specific range, an excellent adhesion prevention effect can be exerted. In addition, if used for the purpose of tendon adhesion prevention, it can effectively improve the joint movable area. As an example of the uncross-linked lysine residue number of the cross-linked gelatin gel used in the adhesion prevention material of the present invention, 1.20×10 ^-4 mol/g~2.50×10 ^-4 mol/g can be cited. Furthermore, from the viewpoint of further enhancing the anti-adhesion effect and the improvement effect of the joint movable area, examples of suitable ranges of the number of uncross-linked lysine residues in the cross-linked gelatin gel used in the anti-adhesion material of the present invention include: 1.20×10 ^-4 mol/g~2.50×10 ^-4 mol/g, 1.30×10 ^-4 mol/g~2.50×10 ^-4 mol/g, 2.10×10 ^-4 mol/g~2.50×10 ^-4 mol/g, 2.25×10 ^-4 mol/g~2.50×10 ^-4 mol/g, 1.20×10 ^-4 mol/g~2.30×10 ^-4 mol/g, 1.20×10 ^-4 mol/g~2.15×10 ^-4 mol/g, 1.30×10 ^-4 mol/g~2.30×10 ^-4 mol/g, 1.30×10 ^-4 mol/g~2.15×10 ^-4 mol/g, 2.25×10 ^-4 mol/g~2.50×10 ^-4 mol/g, 2.05×10 ^-4 mol/g~2.30×10 ^-4 mol/g, 1.15×10 ^-4 mol/g~2. 50×10 ^-4 mol/g, 1.15×10 ^-4 mol/g~2.30×10 ^-4 mol/g, or 1.15×10 ^-4 mol/g~2.15×10 ^-4 mol/g.

In the present invention, the number of uncrosslinked lysine residues of the crosslinked gelatin gel is a value measured according to the following method. <Method for measuring the number of uncrosslinked lysine residues> Prepare a dry powder of the crosslinked gelatin gel to be measured. Add 1 ml of a 4 wt% sodium bicarbonate aqueous solution to 11 mg of the dried crosslinked gelatin powder, and then add 0.5 ml of an N,N-dimethylformamide (DMF) solution containing 1 wt% of 2,4,6-trinitrobenzenesulfonic acid, and stir at 40°C for 4 hours. Then, add 3 ml of 6N hydrochloric acid and stir at 60°C for 2 hours. Thereafter, wash with diethyl ether, and remove the diethyl ether remaining in the washed aqueous solution by reduced pressure distillation with an evaporator. Purified water was added to the obtained aqueous solution to adjust to 50 ml, and the absorbance at 346 nm (optical diameter of the cuvette 1 cm) (absorbance of the sample) was measured with a spectrophotometer. Furthermore, 1 ml of a 4 wt% sodium bicarbonate aqueous solution was added to 11 mg of the dried cross-linked gelatin powder, and then 3 ml of 6N hydrochloric acid was added, followed by 0.5 ml of a 1 wt% N,N-dimethylformamide (DMF) solution containing 2,4,6-trinitrobenzenesulfonic acid, and stirred at 40°C for 4 hours. Then, stirred at 60°C for 2 hours. After that, the solution was washed with diethyl ether, and the diethyl ether remaining in the washed aqueous solution was removed by reduced pressure distillation with an evaporator. Add purified water to the obtained aqueous solution to adjust to 50 ml, and measure the absorbance at 346 nm (optical diameter of cuvette 1 cm) (absorbance of blank sample) with a spectrophotometer. [Formula 1] Number of uncrosslinked lysine residues (mol/g) = {(absorbance of sample - absorbance of blank sample) × 0.05 (l)} / {1.46 × 10 ⁴ (l/mol・cm: absorbance coefficient of N ⁶ -(2,4,6-trinitrophenyl) lysine) × 1 (cm: cuvette length) × 0.011 (g: gelatin weight)}

・Degree of crosslinking As mentioned above, the number of uncrosslinked lysine residues in the crosslinked gelatin gel is a physical property value that serves as an index of the degree of crosslinking, and the number of uncrosslinked lysine residues in the crosslinked gelatin gel is related to the degree of crosslinking. Therefore, the degree of crosslinking of the crosslinked gelatin gel used in the present invention corresponds to the range of the number of uncrosslinked lysine residues mentioned above, but specifically, it can be 30~67%. Examples of suitable ranges of the degree of crosslinking of the crosslinked gelatin gel used in the present invention include 30~39%, 30~34%, 30~62%, 34~62%, 34~39%, 39~62%, 30~67%, 34~67% or 39~67%.

The degree of crosslinking in the crosslinked gelatin gel is the value calculated by measuring the number of uncrosslinked lysine residues of the gelatin before crosslinking, which is the raw material of the crosslinked gelatin gel, using the above method and following the following calculation formula. [Number 2] Degree of crosslinking (%) = {1-(number of uncrosslinked lysine residues of crosslinked gelatin gel (mol/g)/number of lysine residues of gelatin before crosslinking (mol/g))}×100

・Hydrolysis properties The cross-linked gelatin gel used in the present invention is not particularly limited in terms of hydrolysis properties as long as the number of uncross-linked lysine residues described above is satisfied. However, for example, as the hydrolysis properties of the cross-linked gelatin gel used in the present invention caused by the enzyme, the pepsin digestion time measured under the conditions shown below is 90 to 320 minutes. Suitable examples of the range of the pepsin digestion time include 90 to 240 minutes, 90 to 150 minutes, 90 to 120 minutes, 120 to 240 minutes, 120 to 150 minutes, 150 to 240 minutes, 120 to 320 minutes, 120 to 310 minutes, 150 to 320 minutes, or 150 to 310 minutes. By satisfying such a pepsin digestion time, the adhesion prevention effect and the improvement effect of the joint movable area can be further improved. ＜Method for measuring pepsin digestion time＞ Prepare a dry powder of the cross-linked gelatin gel to be measured. Mix 0.02g of the dried cross-linked gelatin powder and 0.08g of the solvent (physiological saline with polyethylene glycol 20000 added to a concentration of 10g/100ml) as a measurement sample. Separately, add pepsin from pig gastric mucosa to a 0.1N hydrochloric acid aqueous solution to dissolve it in a concentration of 1200Units/ml, and filter it to prepare a pepsin solution. 0.1 g of the sample to be measured was added to 40 mL of the pepsin solution, and the mixture was stirred at 37°C in an incubator using a stirrer, while the time until the cross-linked gelatin (insoluble matter) disappeared was determined as the pepsin digestion time. The disappearance of the cross-linked gelatin was observed every 10 minutes from the time when the sample to be measured was added. If the disappearance could not be confirmed by the eye after 360 minutes from the time when the sample to be measured was added, the pepsin digestion time was defined as greater than 360 minutes. In addition, if the insoluble matter could not be confirmed by the eye, it was defined as disappeared. Furthermore, in this manual, 1 unit of pepsin activity refers to the amount of enzyme that increases the absorbance of TCA-soluble substances (free amino acids) at 280 nm in the reaction solution by 0.001 in 1 minute after it acts on hemoglobin at pH 2.0 and 37°C.

・Source of gelatin The source of gelatin used in the formation of the cross-linked gelatin gel used in the present invention is not particularly limited, and may be, for example, any of the following: from mammals such as cows and pigs; from fish such as sharks, tilapias, salmon, catfish, etc. Preferably, it is from mammals, and more preferably, it is from pigs. In addition, gelatin may be obtained by any treatment such as acid treatment, alkali treatment, enzyme treatment, heat treatment, etc.

・Production method The cross-linked gelatin gel used in the present invention can be obtained by cross-linking the gelatin gel so as to satisfy the aforementioned range of the number of uncross-linked lysine residues. The method for cross-linking the gelatin gel is not particularly limited as long as the aforementioned range of the number of uncross-linked lysine residues is satisfied, but for example, it can be any of: thermal cross-linking by heating; electron beam cross-linking by irradiation with electron beams such as ultraviolet rays and far infrared rays; chemical cross-linking by treatment with glutaraldehyde, tannin, alum, aluminum sulfate, succinimidyl poly-L-glutamine, etc.; enzyme cross-linking by treatment with enzymes such as glutaminase, etc. As the crosslinked gelatin used in the present invention, from the viewpoints of biocompatibility and anti-adhesion effect, a crosslinked gelatin gel obtained by thermal crosslinking is preferably cited.

The following is a description of a method for producing a crosslinked gelatin gel by thermal crosslinking.

In the preparation of the cross-linked gelatin gel used in the present invention by heat cross-linking, the dried gelatin gel is heated until the number of uncross-linked lysine residues meets the above range.

The dried gelatin gel to be heated can be obtained by drying a gelatin hydrogel obtained by cooling a gelatin solution. The gelatin content in the gelatin solution may be, for example, 1 to 6% by weight, preferably 2 to 6% by weight, and more preferably 3 to 5% by weight.

The drying of gelatin hydrogel can be carried out by freeze drying, reduced pressure drying, heat drying, etc.

The dried gelatin gel may be subjected to a pulverization process or the like to be powdered as necessary.

The heating conditions for the dried gelatin gel can be set to take into account the shape of the dried gelatin gel and to satisfy the aforementioned range of the number of uncrosslinked lysine residues. The heating temperature can be, for example, 148 to 170°C, preferably 148 to 160°C, and more preferably 148 to 152°C. The heating time can be, for example, more than 16 hours, preferably 18 to 100 hours, and more preferably 18 to 72 hours or 18 to 74 hours. In the production of conventional heat-crosslinked gelatin gel, although a temperature condition of about 140°C or less is adopted, it is difficult to obtain a crosslinked gelatin gel that satisfies the above-mentioned range of the number of uncrosslinked lysine residues under such temperature conditions.

The cross-linked gelatin obtained in this way can also be subjected to treatments such as pulverization and granulation as necessary.

[Form of the anti-adhesion material] In the anti-adhesion material of the present invention, the cross-linked gelatin gel is applied to the affected part in the state of a hydrogel. In one embodiment of the anti-adhesion material of the present invention, the cross-linked gelatin gel is contained in the state of a hydrogel and applied to the affected part as it is. In another embodiment of the anti-adhesion material of the present invention, the cross-linked gelatin gel is contained in the state of an aerogel and is mixed with an aqueous solvent to be used in the state of a hydrogel. In the present invention, aerogel refers to a gel containing air as a dispersion medium (i.e., a dry gel), including: a dry gel obtained by supercritical drying (aerogel in a narrow sense), a dry gel obtained by drying under atmospheric pressure (xerogel), and a dry gel obtained by freeze drying (cryogel). In addition, in the present invention, hydrogel refers to a gel containing an aqueous solvent as a dispersion medium (i.e., a water-containing gel).

In the anti-adhesion material of the present invention, when the cross-linked gelatin gel is provided in the state of a hydrogel, it is sufficient to contain the cross-linked gelatin gel and an aqueous solvent. Examples of the aqueous solvent include physiological saline, purified water, and the like. In the anti-adhesive material of the present invention, when the cross-linked gelatin gel is provided in the state of a hydrogel, the content of the cross-linked gelatin is, for example, 5 to 95% by weight, preferably 10 to 40% by weight, more preferably 14 to 30% by weight, and even more preferably 20 to 25% by weight; the content of the aqueous solvent is, for example, 5 to 95% by weight, preferably 55 to 88% by weight, more preferably 60 to 85% by weight, and even more preferably 65 to 75% by weight.

In the adhesion prevention material of the present invention, when the cross-linked gelatin gel is provided in the state of a hydrogel, the shape of the hydrogel can be, for example, a paste, a sheet, a film, etc. Among them, a paste is preferred. If a paste-like hydrogel is used, the diffusion property in the affected part after application will become good. For example, even if it is administered to the tendon, it can still diffuse into fine and complex tissues and spread throughout the adhesion prevention material affected part. When the paste-like hydrogel is made, the aqueous solvent can be contained in the aggregate of the powdered cross-linked gelatin gel. The average particle size of the powdered crosslinked gelatin gel (in a non-water-absorbed state) used in the preparation of the paste-like hydrogel is, for example, 50 to 10,000 μm, preferably 50 to 500 μm, and more preferably 50 to 150 μm. In the present invention, the average particle size of the powdered aerogel is the particle size (median particle size, _D50 ) at which the cumulative fraction is 50% in the volume cumulative reference particle size distribution measured using a laser analysis-scattering particle size distribution measuring device.

In the anti-adhesion material of the present invention, when the cross-linked gelatin gel is provided in the form of a paste-like hydrogel, it is preferably provided in a syringe so as to be easily applied to the affected area.

Furthermore, in the anti-adhesion material of the present invention, when the cross-linked gelatin gel is provided in the state of an aerogel and used as a hydrogel when used, it is desirable to prepare a two-component type preparation of a first preparation containing the cross-linked gelatin gel and a second preparation containing an aqueous solvent. As an embodiment of such a two-component type preparation, the first preparation and the second preparation are provided by filling separate syringes, and when used, the two syringes are connected and the first preparation and the second preparation are mixed by pumping.

In the adhesion prevention material of the present invention, when the cross-linked gelatin gel is provided in the state of aerogel, the shape of the aerogel can be, for example, powder, sheet, film, etc. Among them, the powder is preferred. If a powdered aerogel is used, it is mixed with an aqueous solvent during use to form a paste, so the diffusion property in the affected area after application will become good. For example, even if it is administered to the tendon, it will still diffuse into fine and complex tissues and can be spread throughout the affected area of the adhesion prevention material. When the cross-linked gelatin gel is provided in the state of powdered aerogel, its average particle size is the same as that of the aforementioned powdered cross-linked gelatin gel (in a non-water-absorbed state).

In the anti-adhesion material of the present invention, when the cross-linked gelatin gel is provided in the state of an aerogel, the type of aqueous solvent mixed during use, the content of the cross-linked gelatin gel and the aqueous solvent in the hydrogel prepared during use, etc. are the same as when the cross-linked gelatin gel is provided in the state of a hydrogel.

[Additives that can be mixed into the anti-adhesion material] The anti-adhesion material of the present invention may also contain polyethylene glycol. The polyethylene glycol used in the present invention is preferably solid at room temperature, and specifically includes: polyethylene glycol 1540, polyethylene glycol 2000, polyethylene glycol 4000, polyethylene glycol 6000, polyethylene glycol 11000, polyethylene glycol 20000, etc. When polyethylene glycol is included in the anti-adhesion material of the present invention, its content is not particularly limited, but for example, it can be 0.1~30% by weight, preferably 1~20% by weight, and more preferably 1~15% by weight.

Furthermore, the anti-adhesion material of the present invention may also contain a surfactant as necessary. The surfactant used in the present invention may be any of non-ionic, cationic, anionic or amphoteric, but as a suitable example, non-ionic surfactants can be cited. As non-ionic surfactants, specifically, polyoxyethylene sorbitan fatty acid esters such as polysorbate 20, polysorbate 60, polysorbate 65, and polysorbate 80; polyoxyethylene hardened castor seed oil, sucrose fatty acid esters, glycerol fatty acid esters, polyoxyethylene polyoxypropylene block copolymers, polyoxyethylene polyoxypropylene glycol, etc. These non-ionic surfactants can be used alone or in combination of two or more. When a surfactant is contained in the anti-adhesion material of the present invention, the content thereof can be appropriately determined according to the type of surfactant used, but can be, for example, 0.1 to 15% by weight, preferably 0.2 to 10% by weight, and more preferably 0.2 to 1% by weight.

Furthermore, the anti-adhesion material of the present invention may also contain a stabilizer as necessary. Specifically, as stabilizers, there can be listed: polyglucose (polyglucose 40, etc.), cyclodextrin, polyvinyl pyrrolidone, glycerol, propylene glycol, sorbitol, hydroxypropyl cellulose, sodium carboxymethyl cellulose, chondroitin sodium sulfate, etc. These stabilizers can be used alone or in combination of two or more. When the stabilizer is contained in the anti-adhesion material of the present invention, its content can be appropriately determined according to the type of surfactant used, but for example, it can be listed as 0.1~20% by weight, preferably 1~15% by weight, and more preferably 4~10% by weight.

Furthermore, the anti-adhesion material of the present invention may also contain an alkali metal salt as necessary. Specifically, the alkali metal salt includes sodium chloride, potassium chloride, etc. Such alkali metal salts may be used alone or in combination of two or more. When the alkali metal salt is contained in the anti-adhesion material of the present invention, its content is not particularly limited, but for example, it may be 0.01 to 5% by weight, preferably 0.1 to 2% by weight, and more preferably 0.5 to 1% by weight.

In addition, the anti-adhesion material of the present invention may also contain additives such as shaping agents, binders, lubricants, pH adjusters, buffers, preservatives, antioxidants, colorants, anti-humidifiers, and oils (such as castor oil) as necessary.

In addition to the aforementioned components, the anti-adhesion material of the present invention may also contain pharmacological components such as urea, antibacterial agents, antibiotics, anti-inflammatory agents, blood circulation improvers, steroids, enzyme inhibitors, growth factors, and various vitamins, as necessary, for the purpose of promoting therapeutic effects and preventing bacterial infections. The anti-adhesion material of the present invention will stay at the affected area for a certain period of time, so by containing the aforementioned pharmacological components, it can be used as a drug delivery system for the purpose of sustained release of pharmacological components.

When the anti-adhesion material of the present invention is provided as a two-dose preparation of a first preparation containing a cross-linked gelatin gel and a second preparation containing an aqueous solvent, these pharmacological components and additives may be contained in either the first preparation or the second preparation, or may be contained in both of them. When the anti-adhesion material of the present invention is provided as a two-dose preparation of a first preparation and a second preparation, the content of the aforementioned additives is the value after mixing the first preparation and the second preparation.

[Application] The adhesion prevention material of the present invention is used in the field of surgery such as abdominal viscera or in the field of plastic surgery for tendons, nerves, and joints to prevent adhesion of biological tissues after surgery such as incision and endoscopic treatment. In particular, among the parts that need to prevent adhesion, tendons (especially tendons in joints) can be cited as a suitable example. If the adhesion prevention material of the present invention is used for the purpose of preventing adhesion of tendons in joints and their surrounding tissues, it can not only prevent adhesion, but also effectively improve the movable area of the joint.

The dosage of the anti-adhesion material of the present invention can be appropriately set according to the state of the target site to prevent adhesion. For example, the anti-adhesion material of the present invention can be set to an amount of about 20 to 120 mg per 1 cm ² of the target site to prevent adhesion, calculated as the dry weight of the cross-linked gelatin gel (converted to the weight of the gelatin itself).

2. Second embodiment of the anti-adhesive material In another embodiment of the anti-adhesive material of the present invention, it is characterized by comprising a cross-linked gelatin gel having a pepsin digestion time of 90 to 320 minutes. In such an embodiment, examples of a suitable range of the pepsin digestion time of the cross-linked gelatin gel include: 90 to 240 minutes, 90 to 150 minutes, 90 to 120 minutes, 120 to 240 minutes, 120 to 150 minutes, 150 to 240 minutes, 120 to 320 minutes, 120 to 310 minutes, 150 to 320 minutes, or 150 to 310 minutes. The pepsin digestion time of the cross-linked gelatin gel in such an embodiment is as described in the column of "1. The first embodiment of the anti-adhesion material". In addition, although there are no special restrictions on the cross-linked gelatin gel used in such an embodiment, the number of uncross-linked lysine residues, the degree of cross-linking, the source of gelatin, the method of manufacturing the cross-linked gelatin, etc., the specific scope example is the same as the case of the aforementioned "1. The first embodiment of the anti-adhesion material". In such an embodiment, the form of the anti-adhesion material, other components that can be blended into the anti-adhesion material, the use of the anti-adhesion material, etc. are also the same as the case of the aforementioned "1. The first embodiment of the anti-adhesion material". Implementation example

Although the present invention is described in detail below based on embodiments and the like, the present invention is not limited thereto and the like.

Production example: Production of cross-linked gelatin powder and measurement of physical properties 1. Production of cross-linked gelatin powder [Production examples 1~7 and 9~12] 4.5 wt% of gelatin (HMG-BP MediGelatin, Nichibei Co., Ltd.) was dissolved in purified water to prepare a gelatin solution. After the gelatin solution was cooled to gel, the gel was frozen at -37°C and freeze-dried. The freeze-drying conditions were: the primary drying was set to -10°C, the secondary drying was set to 10°C, and the final drying was set to 30°C. From the primary drying to the final drying, it was carried out under reduced pressure. After freeze drying, cut into about 5 mm squares with scissors. After cutting, the powder was crushed into a particle size of about 10-500 μm using an ultracentrifugal grinder (ZM200, Verder Scientific Co., Ltd.), and a sieve was used to recover only powder with a particle size of 53-150 μm, and the average particle size was measured. The recovered powder was thermally crosslinked using an air supply constant temperature dryer (WFO-420W, Tokyo Rikki Co., Ltd.) under the conditions shown in Table 1 to obtain crosslinked gelatin powder. Furthermore, the average particle size of the powder before the heat crosslinking treatment is the median particle size measured using a laser analysis-scattering particle size distribution measuring device ("LS 13 320", Beckman Coulter Co., Ltd.) in Production Examples 1 to 7, 11 and 12, and the number-based arithmetic mean length diameter measured using an image analysis device ("MorphologiG3S", Spectris Co., Ltd.) in Production Examples 9 and 10.

[Production Example 8] 2.5 wt% of gelatin (RM-100, Jellice Co., Ltd.) was dissolved in distilled water, stirred and mixed with a homogenizer for 30 seconds and pre-frozen (-40°C, 12 hours), and then freeze-dried (FZ-COMPACT, LABOCONCO) (about -45°C, 40 hours) to obtain a gelatin sponge. The obtained gelatin sponge was thermally crosslinked (140 degrees, 48 hours) using a constant temperature dryer (DO-300PA, AS ONE Co., Ltd.), and then crushed into a particle size of less than 500μm using an ultracentrifugal grinder (ZM-100, Retsch Co., Ltd.), and only powder with a particle size of 250~500μm was recovered using a sieve. The average particle size (number-based arithmetic mean length diameter) was measured using an image analysis device ("MorphologiG3S", Spectris Co., Ltd.).

2. Measurement of the number of uncrosslinked lysine residues and degree of crosslinking 11 mg of crosslinked gelatin powder and gelatin powder before crosslinking were weighed and 1 mL of 4 wt% sodium bicarbonate aqueous solution was added. 0.5 mL of N,N-dimethylformamide (DMF) solution (Tokyo Chemical Industry Co., Ltd.) containing 1 wt% of 2,4,6-trinitrobenzenesulfonic acid was added to this solution and stirred at 40°C for 4 hours. In addition, 3 mL of 6N hydrochloric acid was added and stirred at 60°C for 2 hours. After that, diethyl ether was washed with the solution and the diethyl ether remaining in the washed aqueous solution was removed by reduced pressure distillation in an evaporator. Purified water was added to the obtained aqueous solution to adjust to 50 mL, and the absorbance at 346 nm (optical diameter of the spectrophotometer was 1 cm) (absorbance of the sample) was measured using a spectrophotometer (U-3900, Hitachi High-tech Science).

Furthermore, 1 mL of a 4 wt% sodium bicarbonate aqueous solution was added to 11 mg of each of the dried crosslinked gelatin powder and the gelatin powder before crosslinking, and then 3 mL of 6N hydrochloric acid was added, followed by 0.5 mL of a 1 wt% N,N-dimethylformamide (DMF) solution containing 2,4,6-trinitrobenzenesulfonic acid, and stirred at 40°C for 4 hours. Then, stirred at 60°C for 2 hours. After that, the solution was washed with diethyl ether, and the diethyl ether remaining in the washed aqueous solution was removed by reduced pressure distillation with an evaporator. Purified water was added to the obtained aqueous solution to adjust the volume to 50 mL, and the absorbance at 346 nm (optical diameter of the spectrophotometer tube was 1 cm) was measured using a spectrophotometer (U-3900, Hitachi High-tech Science Co., Ltd.) (absorbance of the blank sample).

The following formula was used to calculate the number of uncrosslinked lysine residues for the gelatin powder before crosslinking and the crosslinked gelatin powder. [Equation 3] Number of uncrosslinked lysine residues (g/mol) = {(absorbance of sample - absorbance of blank sample) × 0.05 (l)} / {1.46 × 10 ⁴ (l/mol・cm) × 1 (cm) × 0.011 (g)}

In addition, the crosslinking degree of the crosslinked gelatin powder was calculated according to the following formula. [Formula 4] Crosslinking degree (%) = {1-(Number of uncrosslinked lysine residues in the crosslinked gelatin powder (mol/g)/Number of lysine residues in the gelatin powder before crosslinking (mol/g))}×100

3. Measurement of pepsin digestion time 0.02 g of dried cross-linked gelatin powder and 0.08 g of solvent (physiological saline solution to which polyethylene glycol 20000 was added to give a concentration of 10 g/100 mL) were mixed to prepare a sample for measurement. Separately, pepsin from pig gastric mucosa ("P7125", Sigma Aldrich) was added to a 0.1 N hydrochloric acid aqueous solution to dissolve it at a concentration of 1200 units/mL, and the solution was filtered to prepare a pepsin solution. 0.1 g of the sample for measurement was added to 40 mL of the pepsin solution, and the mixture was stirred in an incubator at 37°C using a stirrer. The time until the cross-linked gelatin (insoluble matter) disappeared was determined as the pepsin digestion time. If the insoluble matter cannot be confirmed by visual inspection, it is considered to have disappeared.

4. Measurement results of cross-linked gelatin powder The manufacturing conditions, number of uncross-linked lysine residues, degree of cross-linking and pepsin digestion time of each cross-linked gelatin powder are shown in Table 1.

[Table 1] Manufacturing example 1 2 3 4 5 6 7 8 9 10 11 12 Manufacturing conditions Gelatin used as raw material ^# A A A A A A A B A A A A Average particle size of gelatin powder subjected to thermal crosslinking treatment (μm) 106 120 112 106 88 106 88 622 127 94 106 100 Crosslinking temperature (℃) 150 150 150 150 150 150 150 140 150 150 150 150 Crosslinking time (h) 3 12 18 twenty four twenty four 72 72 48 25 25 74 25 Measurement results Number of uncross-linked lysine residues (×10 ^-4 mol/g) 3.11 2.60 2.49 2.28 2.10 1.31 1.06 2.55 2.36 2.32 1.15 2.30 Number of cross-linked lysine residues (×10 ^-4 mol/g) 0.34 0.91 1.07 1.17 1.32 2.14 2.36 0.82 1.12 1.18 2.37 1.25 Number of lysine residues before cross-linking (×10 ^-4 mol/g) 3.45 3.51 3.55 3.45 3.42 3.45 3.42 3.37 3.48 3.50 3.52 3.55 Crosslinking degree (%) 10 26 30 34 39 62 69 twenty four 32 34 67 35 Pepsin digestion time (min) 30 40 90 120 150 240 >360 80 100 90 310 100 # A stands for "HMG-BP MediGelatin Gelatin" (Ripi Co., Ltd.), and B stands for "RM-100" (Jellice Co., Ltd.).

Test example (verification of anti-adhesion effect) 1. Test example 1 1-1. Preparation of sample Prepare a sample by mixing the components shown in Table 2.

[Table 2] Comparative Example 1-1 (Control Group) Comparison Example 1-2 Preparation Example 1 Cross-linked Gelatin Powder - 20 Polyethylene glycol 20000 (Fuji Film Wako Pure Chemical Industries, Ltd.) 10 8 Physiological saline solution 90 72 Total (weight %) 100 100 Characteristics Liquid Paste

1-2. Experimental method As the feed animals, 10-week-old rabbits (Kbl: JW, male) (10 per group) were used. As anesthetics, ketamine (trade name "KETALAR intramuscular injection 500 mg", Daiichi Sankyo Propharma Co., Ltd.) and xylazine (trade name "Selactar 2% injection", Bayer Pharmaceuticals Co., Ltd.) were mixed at a ratio of 80 liquid volumes of ketamine to 20 liquid volumes of xylazine, and 5 mL/body of the mixture was injected intramuscularly into the rabbits. Next, 0.3 mL/body of butylorphan (trade name "Lepetan injection 0.2 mg", Otsuka Pharmaceutical Co., Ltd.) was injected intramuscularly as an analgesic, and 0.4 mL/body of enrofloxacin (Baytril 2.5% injection, Bayer Pharmaceuticals Co., Ltd.) was injected subcutaneously as an antibacterial drug.

After the hair of the left hind limb of the anesthetized animal was removed, the extension and flexion of the toe to be treated were immobilized. That is, the animal was fixed in a supine position on a fixator (trade name "Kitajima Fixator", Natsume Seisakusho Co., Ltd.), and after disinfecting the surgical area, i.e., the area around the nail, the extensor digitorum longus tendon distal to the inferior extensor retinaculum was cut off by about 5 mm to control spontaneous extension. Then, the animal was fixed in a ventral position on the fixator, and after disinfecting the surgical area, i.e., the area around the heel, the superficial flexor digitorum tendon and the deep flexor digitorum tendon were cut off by about 5 mm each to control spontaneous flexion. After excision of the extensor and flexor tendons, the skin was closed with single-knot sutures using 3-0 resistant suture (trade name: "resistant suture with needle MM15 3-0N, Matsuda Medical Industry Co., Ltd.").

After immobilization, the animal was fixed in a ventral position on a fixator. After disinfection, the skin of the upper part of the middle toe of the left hind limb (the skin from the MP joint to the PIP joint) was longitudinally incised to expose the flexor tendon. The deep flexor tendon of the digitorum was sharply cut with a scalpel within the exposed flexor tendon. Thereafter, the cut site of the deep flexor tendon of the digitorum was repaired by continuous suture using 6-0 PROLENE (trade name "PROLENE ^R (M8307)", J.S. Co., Ltd.), Kessler grip suture using 5-0 PROLENE (trade name "PROLENE ^R (M8557)", J.S. Co., Ltd.), and horizontal mattress suture. After hemostasis and washing with physiological saline, 0.15 mL of the sample was injected into the exposed deep flexor tendon and its surroundings. Then, the incision was sutured with suture thread (trade name "Palm Sharp Needle, 9mm 3/8, 5-0 Black Nylon", Matsuda Medical Industry Co., Ltd.). In order to prevent self-harm behavior, a collar was wrapped around the head for breeding.

The assessment of the movable area of the toe joint was performed by the following procedure. Two weeks after the start of the specimen administration, 1.45 mL/kg of body weight of a ketamine-xylazine mixed anesthetic solution was administered into the thigh muscle of the animal, and then the carotid artery was cut to bleed. Thereafter, the left hind limb was cut between the talus and the tibia, and the sampled left hind limb was placed on a measuring table. Next, a knot was tied to the deep flexor tendon of the toe joint that had undergone MP joint proximal surgery, and the thread was connected to a tensile testing machine by a hook. In addition, the tensile testing machine is composed of a load measuring device (trade name "Digital Dynamometer ZTS-20N", IMADA Co., Ltd.) and a measuring table (trade name "Horizontal Electric Measuring Table MH2-500", IMADA Co., Ltd.). Therefore, a load of 3.0N was applied to the tendon (usually the minimum force for maximum flexion of the PIP joint and DIP joint in the toes), and photographs were taken before and after the load. The angles of the PIP joint and the DIP joint were measured on the printouts of the photographs, and the movable area of the toe joint of the operated toe was calculated by the following calculation formula. The mean ± standard deviation was calculated for each group, and the significant difference test was performed by an uncorrelated t-test (both sides). [Figure 5] Movement area of toe joint (°) = after load (flexion angle of PIP joint + flexion angle of DIP joint) - before load (flexion angle of PIP joint + flexion angle of DIP joint)

In addition, the assessment of adhesion degree was performed by the following procedure. After the photographs of the movable area of the toe joint were taken, the surgical site was cut open again, and the adhesion degree was assessed according to the following assessment criteria for the three parts of the flexor digitorum profundus tendon and sliding base plate, the flexor digitorum profundus tendon and the flexor digitorum superficialis tendon, and the flexor digitorum profundus tendon and other surrounding tissues. The sum of the scores of the three parts was taken as the individual value. The mean and median values were calculated for each group, and the significant difference test was performed by an uncorrelated t test (two sides). <Evaluation criteria> 0: No adhesion 1: Can be easily peeled off from the surrounding tissue by blunt peeling 2: Can be peeled off from the surrounding tissue by blunt peeling 3: Can be peeled off from the surrounding tissue by blunt peeling although with difficulty 4: Peeling from the surrounding tissue requires sharp peeling

1-3. Test results The obtained results are shown in FIG1. As a result, the sample of Comparative Example 1-2 could not confirm the reduction of the adhesion score, nor could it confirm the effect of expanding the movable area of the toe joint. Furthermore, in the surgical site 2 weeks after the sample was administered, no sample residue was confirmed in Comparative Example 1-1, while the sample residue rate in Comparative Example 1-2 was about 70%. That is, from this result, it can be confirmed that the cross-linked gelatin in Production Example 1 (uncross-linked lysine residue number of 3.11× ^10-4 mol/g, cross-linking degree of 10%, pepsin digestion time of 30 minutes) does not have satisfactory performance as an anti-adhesion material.

2. Test Example 2 2-1. Preparation of Samples Prepare the samples by mixing the ingredients shown in Table 3.

[Table 3] Comparative Example 2-1 (Control Group) Comparison Example 2-2 Preparation Example 2 Crosslinked Gelatin Powder - 20 Polyethylene glycol 20000 (Fuji Film Wako Pure Chemical Industries, Ltd.) 10 8 Physiological saline 90 72 Total (weight %) 100 100 Characteristics Liquid Paste

2-2. Test method The anti-sticking effect was evaluated by the same method as in the above-mentioned Test Example 1. In addition, in Comparative Example 2-1, 10 rabbits were used for the test, and in Comparative Example 2-2, 8 rabbits were used for the test.

2-3. Test results The obtained results are shown in FIG2. As a result, although the sample of Comparative Example 2-2 was confirmed to have an effect of expanding the movable area of the toe joint, the reduction of the adhesion score could not be confirmed. Furthermore, in the surgical site 2 weeks after the sample was administered, no sample residue was confirmed in Comparative Example 2-1, while the residual rate of the sample of Comparative Example 2-2 was about 100%. That is, from this result, it can be confirmed that the cross-linked gelatin in Production Example 2 (uncross-linked lysine residue number of 2.60× ^10-4 mol/g, cross-linking degree of 26%, pepsin digestion time of 40 minutes) does not have satisfactory performance as an anti-adhesion material.

3. Test Example 3 3-1. Preparation of Samples Prepare the samples by mixing the ingredients shown in Table 4.

[Table 4] Comparative Example 3-1 (Control Group) Example 3-1 Example 3-2 Preparation Example 4 Cross-linked Gelatin Powder - 20 - Preparation Example 6 Cross-linked Gelatin Powder - - 20 Polyethylene glycol 20000 (Fuji Film Wako Pure Chemical Industries, Ltd.) 10 8 8 Physiological saline solution 90 72 72 Total (weight %) 100 100 100 Characteristics Liquid Paste Paste

3-2. Test method The anti-sticking effect was evaluated by the same method as in the above-mentioned Test Example 1, except that the significant difference test was set to Dunnett's multiple comparison (two sides). In addition, the test was conducted on 10 rabbits in Comparative Example 3-1, on 10 rabbits in Example 3-1, and on 9 rabbits in Example 3-2.

3-3. Test results The obtained results are shown in FIG3. As a result, the movable area of the toe joint of the samples of Examples 3-1 and 3-2 was significantly increased and the adhesion score was significantly reduced compared to that of Comparative Example 3-1. Furthermore, in the surgical site 2 weeks after the sample administration, no sample residue was confirmed in Comparative Example 3-1, while the residual rate of the samples of Examples 3-1 and 3-2 was about 100%. That is, from the present results, it can be confirmed that the cross-linked gelatin of Production Example 4 (the number of uncross-linked lysine residues is 2.28× ^10-4 mol/g, the degree of cross-linking is 34%, and the pepsin digestion time is 120 minutes) and the cross-linked gelatin of Production Example 6 (the number of uncross-linked lysine residues is 1.31× ^10-4 mol/g, the degree of cross-linking is 62%, and the pepsin digestion time is 240 minutes) have satisfactory performance as anti-adhesion materials.

4. Test Example 4 4-1. Preparation of Samples Prepare the samples by mixing the components shown in Table 5.

[Table 5] Comparative Example 4-1 (Control Group) Example 4-1 Comparison Example 4-2 Preparation Example 5 Crosslinked Gelatin Powder - 20 - Crosslinked gelatin powder of Production Example 7 - - 20 Polyethylene glycol 20000 (Fuji Film Wako Pure Chemical Industries, Ltd.) 10 8 8 Physiological saline 90 72 72 Total (weight %) 100 100 100 Characteristics Liquid Paste Paste

4-2. Test method The anti-sticking effect was evaluated by the same method as in the above-mentioned Test Example 1, except that the significant difference test was set to Dunnett's multiple comparison (two sides). In addition, in Comparative Example 4-1, 8 rabbits were used for the test, in Comparative Example 4-2, 10 rabbits were used for the test, and in Example 4-1, 9 rabbits were used for the test.

4-3. Test results The obtained results are shown in FIG4. As a result, although the sample of Comparative Example 4-2 confirmed the effect of expanding the movable area of the toe joint, the reduction of the adhesion score was not confirmed. On the other hand, in the sample of Example 4-1, the movable area of the toe joint was significantly increased, and the adhesion score was also significantly reduced. Furthermore, in the surgical site 2 weeks after the sample was injected, the residual sample of Comparative Example 4-1 could not be confirmed, and the residual rate of the samples of Comparative Example 4-2 and Example 4-1 was about 100%. That is, it can be confirmed from the present results that, although the cross-linked gelatin of Production Example 5 (the number of uncross-linked lysine residues is 2.10× ^10-4 mol/g, the degree of cross-linking is 39%, and the pepsin digestion time is 150 minutes) has satisfactory performance as an anti-adhesive material, the cross-linked gelatin of Production Example 7 (the number of uncross-linked lysine residues is 1.06× ^10-4 mol/g, the degree of cross-linking is 69%, and the pepsin digestion time is greater than 360 minutes) does not have satisfactory performance as an anti-adhesive material.

5. Test Example 5 5-1. Preparation of Samples Prepare the samples by mixing the ingredients shown in Table 6.

[Table 6] Comparative Example 5-1 (Control Group) Example 5-1 Preparation Example 3 Cross-linked Gelatin Powder - 20 Polyethylene glycol 20000 (Fuji Film Wako Pure Chemical Industries, Ltd.) 10 8 Physiological saline solution 90 72 Total (weight %) 100 100 Characteristics Liquid Paste

5-2. Evaluation of anti-sticking effect The anti-sticking effect was evaluated by the same method as in the above-mentioned Test Example 1. In addition, 7 rabbits were used for the test in both Comparative Example 5-1 and Example 5-1.

5-3. Test results The obtained results are shown in FIG5. As a result, in the sample of Example 5-1, the movable area of the toe joint was significantly increased, and the adhesion score was also significantly reduced. Furthermore, in the surgical site 2 weeks after the sample was administered, no sample residue was confirmed in Comparative Example 5-1, while the residual rate of the sample of Example 5-1 was about 100%. That is, from this result, it can be confirmed that the cross-linked gelatin of Production Example 3 (uncross-linked lysine residue number of 2.49× ^10-4 mol/g, cross-linking degree of 30%, pepsin digestion time of 90 minutes) has satisfactory performance as an anti-adhesion material.

6. Test Example 6 6-1. Preparation of Samples Prepare the samples by mixing the ingredients shown in Table 7.

[Table 7] Comparative Example 6-1 (Control Group) Comparison Example 6-2 Crosslinked Gelatin Powder of Production Example 8 - 20 Physiological saline 100 80 Total (weight %) 100 100 Characteristics Liquid Paste

6-2. Evaluation of anti-sticking effect The anti-sticking effect was evaluated by the same method as in the above-mentioned test example 1. In addition, in comparative example 6-1, 7 rabbits were used for the test, and in comparative example 6-2, 5 rabbits were used for the test.

6-3. Test results The obtained results are shown in FIG6. In the sample of Comparative Example 6-2, no significant difference was confirmed in both the adhesion score and the movable area of the toe joint compared with Comparative Example 6-1. Furthermore, in the surgical site 2 weeks after the sample was administered, no residual sample was confirmed in Comparative Example 6-1, while the residual rate of the sample of Comparative Example 6-2 was about 100%. That is, from this result, it can be confirmed that the cross-linked gelatin of Production Example 8 (uncross-linked lysine residue number is 2.55× ^10-4 mol/g, cross-linking degree is 24%, and pepsin digestion time is 80 minutes) does not have satisfactory performance as an anti-adhesion material.

7. Test Example 7 7-1. Preparation of Samples Prepare the samples by mixing the ingredients shown in Table 8.

[Table 8] Comparative Example 7-1 (Control Group) Example 7-1 Example 7-2 Example 7-3 Preparation Example 9 Crosslinked Gelatin Powder - 20 20 20 Polysorbate 80 (Surfactant Industries, Ltd., Japan) - - 0.4 - Polyglucose 40 (Mingtang Industrial Co., Ltd.) - - - 8 Physiological saline 100 80 79.6 72 Total (weight %) 100 100 100 100 Characteristics Liquid Paste Paste Paste

7-2. Evaluation of adhesion prevention effect The adhesion prevention effect was evaluated by the same method as in Experimental Example 1, except that (1) 11-week-old rabbits (Kbl: JW, male) were used; (2) the dose of the test sample was changed to 0.30 mL; (3) Ravonal (Ravonal injection 0.5 g, Nipro ES Pharma Co., Ltd.) 3.0 mL/body was administered to replace the ketamine-xylazine mixed anesthetic solution when evaluating the movable area of the toe joint; and (4) the significant difference test was set to Dunkley's multiple comparison (both sides). In addition, the experiments were conducted on 9 rabbits in Comparative Example 7-1, Examples 7-1 and 7-2, and on 8 rabbits in Example 7-3.

7-3. Test results The obtained results are shown in Figure 7. As a result, the movable area of the toe joint of the samples of Examples 7-1, 7-2 and 7-3 was significantly increased and the adhesion score was significantly reduced compared to that of Comparative Example 7-1. Furthermore, in the surgical site 2 weeks after the sample was administered, no sample residue was confirmed in Comparative Example 7-1, while the residual rate of the samples of Examples 7-1, 7-2 and 7-3 was about 100%. That is, it can be confirmed from this result that the presence or absence of additives and their types have no effect on the anti-adhesion effect of crosslinked gelatin.

8. Test Example 8 8-1. Preparation of Samples Prepare the samples by mixing the ingredients shown in Table 9.

[Table 9] Comparative Example 8-1 (Control Group) Example 8-1 Example 8-2 Crosslinked Gelatin Powder of Preparation Example 10 - 15 25 Polysorbate 80 (Surfactant Industries, Ltd., Japan) 0.5 0.425 0.375 Physiological saline 99.5 84.575 74.625 Total (weight %) 100 100 100 Characteristics Liquid Paste Paste

8-2. Evaluation of adhesion prevention effect The adhesion prevention effect was evaluated by the same method as in Experimental Example 1, except that (1) 12-week-old rabbits (Kbl: JW, male) were used; (2) the dose of the test sample was changed to 0.30 mL; (3) 3.0 mL/body of Namonal (Namonal injection 0.5 g, Nipro ES Pharma Co., Ltd.) was administered to replace the ketamine-xylazine mixed anesthetic solution when evaluating the movable area of the toe joint; and (4) the significant difference test was set to Dunkley's multiple comparison (both sides). In addition, Comparative Example 8-1, Examples 8-1 and 8-2 were all tested on 8 rabbits.

8-3. Test results The obtained results are shown in FIG8. As a result, the movable area of the toe joint of the samples of Examples 8-1 and 8-2 was significantly increased and the adhesion score was significantly reduced compared to that of Comparative Example 8-1. In addition, the movable area of the toe joint of the sample of Example 8-2 with a high content of crosslinked gelatin was increased and the adhesion score was reduced compared to that of Example 8-1. Furthermore, in the surgical site 2 weeks after the sample was administered, no residual sample was confirmed in Comparative Example 8-1, while the residual rate of the samples of Examples 8-1 and 8-2 was about 100%. That is, it can be confirmed from this result that as the content of cross-linked gelatin in the sample increases, the anti-adhesion effect also increases.

9. Test Example 9 9-1. Preparation of Samples Prepare the samples by mixing the ingredients shown in Table 10.

[Table 10] Comparative Example 9-1 (Control Group) Example 9-1 Crosslinked Gelatin Powder of Preparation Example 11 - 20 Polyglucose 40 (Mingtang Industrial Co., Ltd.) 10 8 Sodium chloride (Ningmen Salt Industry Co., Ltd.) 0.9 0.72 Water for injection 89.1 71.28 Total (weight %) 100 100 Characteristics Liquid Paste

9-2. Evaluation of adhesion prevention effect The adhesion prevention effect was evaluated by the same method as in Experimental Example 1, except that (1) 11-week-old rabbits (Kbl: JW, male) were used; (2) immobilization was performed by the following procedure; (3) the dose of the sample was changed to 0.30 mL; and (4) 3.0 mL/body of Namonal (Namonal injection 0.5 g, Nipro ES Pharma Co., Ltd.) was administered to replace the ketamine-xylazine mixed anesthetic solution when evaluating the movable area of the toe joint. In addition, the test was conducted on 7 rabbits in Comparative Example 9-1, and on 8 rabbits in Example 9-1. <Immobilization procedure> The animal was fixed in a ventral position on a fixator (trade name "Kitajima fixator", Natsume Manufacturing Co., Ltd.), and after disinfecting the surgical area, i.e. the heel area, the deep flexor tendon of the digitorum was cut off by about 5 mm to control spontaneous flexion. After the deep flexor tendon of the digitorum was cut off, the skin was closed with a single knot suture of 3-0 resistant suture (trade name "Attached needle resistant suture MM15 3-0N, Matsuda Medical Industry Co., Ltd.").

9-3. Test results The obtained results are shown in FIG9. As a result, the movable area of the toe joint of the sample of Example 9-1 was significantly increased compared with that of Comparative Example 9-1, and the adhesion score was also significantly reduced. Furthermore, in the surgical site 2 weeks after the sample was administered, the residual rate of the sample of Comparative Example 9-1 was about 29%, while that of the sample of Example 9-1 was about 100%. That is, from this result, it can be confirmed that the cross-linked gelatin of Production Example 11 (the number of uncross-linked lysine residues is 1.15× ^10-4 mol/g, the degree of cross-linking is 67%, and the pepsin digestion time is 310 minutes) has satisfactory performance as an anti-adhesion material.

10. Test Example 10 10-1. Preparation of specimens First, prepare two pre-filled syringes ("ClearJect 2.25mL LL T4", Dacheng Chemical Co., Ltd.). The pre-filled syringe is composed of a barrel with a tip cap, a piston, a plunger rod, and a finger clip. Prepare a solution (the second preparation) by dissolving polyglucose 40 and sodium chloride in water for injection in advance, and fill the second preparation into one of the pre-filled syringes. Fill the pre-filled syringe with a specified amount of cross-linked gelatin powder (the first preparation). Immediately before administration, the prefilled syringe filled with the first preparation and the prefilled syringe filled with the second preparation were connected with a connector ("Luer-fitted soft tube, VRFC6", Nordson MEDICAL), and mixed by pumping (more than 20 reciprocating strokes) to obtain the sample composition shown in Table 11.

[Table 11] Comparative Example 10-1 (Control Group) Example 10-1 Example 10-2 Example 10-3 Crosslinked gelatin powder of Preparation Example 12 - 20 twenty three 25 Polyglucose 40 (Mingtang Industrial Co., Ltd.) 10 8 7.7 7.5 Sodium chloride (Ningmen Salt Industry Co., Ltd.) 0.9 0.72 0.693 0.675 Water for injection 89.1 71.28 68.607 66.825 Total (weight %) 100 100 100 100 Characteristics Liquid Paste Paste Paste

10-2. Evaluation of adhesion prevention effect The adhesion prevention effect was evaluated by the same method as in Experimental Example 1, except that (1) 11-12 week-old rabbits (Kbl: JW, male) were used; (2) immobilization was performed by the same procedure as in Experimental Example 9; (3) the dose of the test sample was changed to 0.30 mL; (4) 3.0 mL/body of Namonal (Namonal injection 0.5 g, Nipro ES Pharma Co., Ltd.) was administered to replace the ketamine-xylazine mixed anesthetic solution when evaluating the movable area of the toe joint; and (5) the significant difference test was set to Dunkley's multiple comparison (both sides). In addition, Comparative Example 10-1 and Examples 10-1 to 10-3 were tested on 8 rabbits.

10-3. Test results The obtained results are shown in FIG10. As a result, the movable area of the toe joint of the samples of Examples 10-1, 10-2 and 10-3 was significantly increased and the adhesion score was significantly reduced compared to Comparative Example 10-1. Furthermore, in the surgical site 2 weeks after the sample was administered, no sample residue was confirmed in Comparative Example 10-1, while the residual rate of the samples of Examples 10-1, 10-2 and 10-3 was about 100%. That is, from this result, it can be confirmed that even in the hydrogel prepared by pumping mixing, it still has satisfactory performance as an anti-adhesion material.

11. Investigation The results of Test Examples 1 to 10 are summarized in Table 12. From these results, it can be confirmed that by selecting a cross-linked gelatin having an uncross-linked lysine residue number of 1.15×10 ^-4 mol/g to 2.50×10 ^-4 mol/g (cross-linking degree of 30 to 62%) or a pepsin digestion time of 90 to 320 minutes, and using it as an anti-adhesion material, a particularly excellent anti-adhesion effect (enlargement effect of the movable area of the toe joint, reduction effect of the adhesion fraction) can be exerted.

[Table 12] Comparison Example 1-2 Comparison Example 6-2 Comparison Example 2-2 Example 5-1 Examples 7-1 to 7-3 Embodiment 8-1~8-2 Embodiment 10-1~10-3 Example 3-1 Example 4-1 Example 3-2 Example 9-1 Comparison Example 4-2 Crosslinked gelatin used Number of uncross-linked lysine residues (×10 ^-4 mol/g) 3.11 2.55 2.60 2.49 2.36 2.32 2.30 2.28 2.10 1.31 1.15 1.06 Number of cross-linked lysine residues (×10 ^-4 mol/g) 0.34 0.82 0.91 1.07 1.12 1.18 1.25 1.17 1.32 2.14 2.37 2.36 Number of lysine residues before cross-linking (×10 ^-4 mol/g) 3.45 3.37 3.51 3.55 3.48 3.50 3.55 3.45 3.42 3.45 3.52 3.42 Crosslinking degree (%) 10 twenty four 26 30 32 34 35 34 39 62 67 69 Pepsin digestion time (min) 30 80 40 90 100 90 100 120 150 240 310 >360 Anti-sticking effect Toe joint movable area (difference from control group) No significant difference No significant difference Significantly expanded Significantly expanded Significantly expanded Significantly expanded Significantly expanded Significantly expanded Significantly expanded Significantly expanded Significantly expanded Significantly expanded Adhesion score (difference from control group) No significant difference No significant difference No significant difference Significantly reduced Significantly reduced Significantly reduced Significantly reduced Significantly reduced Significantly reduced Significantly reduced Significantly reduced No significant difference Survival rate of the specimens in the surgical department 2 weeks after specimen administration 70% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100%

without

[Figure 1] is a graph showing the results of measuring the movable area and adhesion score of the toe joint in Test Example 1. [Figure 2] is a graph showing the results of measuring the movable area and adhesion score of the toe joint in Test Example 2. [Figure 3] is a graph showing the results of measuring the movable area and adhesion score of the toe joint in Test Example 3. [Figure 4] is a graph showing the results of measuring the movable area and adhesion score of the toe joint in Test Example 4. [Figure 5] is a graph showing the results of measuring the movable area and adhesion score of the toe joint in Test Example 5. [Figure 6] is a graph showing the results of measuring the movable area and adhesion score of the toe joint in Test Example 6. [Figure 7] is a graph showing the results of measuring the movable area and adhesion score of the toe joint in Test Example 7. [Figure 8] is a graph showing the results of measuring the movable area and adhesion score of the toe joint in Test Example 8. [Figure 9] is a graph showing the results of measuring the movable area and adhesion score of the toe joint in Test Example 9. [Figure 10] is a graph showing the results of measuring the movable area and adhesion score of the toe joint in Test Example 10.

Claims (9)

An anti-adhesion material comprises a cross-linked gelatin gel having an uncross-linked lysine residue number of 1.15×10 ^-4 mol/g to 2.50×10 ^-4 mol/g. For the anti-adhesion material of claim 1, the pepsin digestion time of the cross-linked gelatin gel is 90 to 320 minutes as measured by the following measurement method: ＜Measurement method of pepsin digestion time＞ Prepare dry powder of cross-linked gelatin to be measured; mix 0.02g of dried cross-linked gelatin powder and 0.08g of solvent (physiological saline with polyethylene glycol 20000 added to 10g/100ml) to prepare a sample for measurement; separately, add pepsin from pig gastric mucosa to 0.1N hydrochloric acid aqueous solution to dissolve it at 1200Units/ml, and filter it to prepare a pepsin solution; add 0.1g of the sample to be measured to 40mL of the pepsin solution, stir it in an incubator at 37℃ with a stirrer, and calculate the time until the cross-linked gelatin (insoluble matter) disappears as the pepsin digestion time. The anti-adhesion material of claim 1 or 2, wherein the cross-linked gelatin gel is a hydrogel. For example, in the anti-sticking material of claim 3, the hydrogel is in a paste form. The anti-adhesion material of claim 1 or 2, wherein the cross-linked gelatin gel is an aerosol gel, and the anti-adhesion material is a two-component type preparation, wherein the two-component type preparation contains a first preparation comprising the cross-linked gelatin gel and a second preparation comprising an aqueous solvent. The anti-adhesion material of claim 1 or 2 is used to prevent adhesion of tendons. A cross-linked gelatin gel having an uncross-linked lysine residue number of 1.15×10 ^-4 mol/g to 2.50×10 ^-4 mol/g is used for the manufacture of anti-adhesion materials. An anti-adhesive material comprising a cross-linked gelatin gel having a pepsin digestion time of 90 to 320 minutes as measured by the following measurement method: ＜Measurement method of pepsin digestion time＞ Prepare dry powder of cross-linked gelatin to be measured; mix 0.02g of dried cross-linked gelatin powder and 0.08g of solvent (physiological saline with polyethylene glycol 20000 added to 10g/100ml) to prepare a sample for measurement; separately, add pepsin from pig gastric mucosa to 0.1N hydrochloric acid aqueous solution to dissolve it at 1200Units/ml, and filter it to prepare a pepsin solution; add 0.1g of the sample to be measured to 40mL of the pepsin solution, stir it in an incubator at 37℃ with a stirrer, and calculate the time until the cross-linked gelatin (insoluble matter) disappears as the pepsin digestion time. A use of a cross-linked gelatin gel having a pepsin digestion time of 90 to 320 minutes as determined by the following measurement method, for the manufacture of an anti-adhesion material: ＜Measurement method of pepsin digestion time＞ Prepare dry powder of cross-linked gelatin to be measured; mix 0.02g of dried cross-linked gelatin powder and 0.08g of solvent (physiological saline with polyethylene glycol 20000 added to 10g/100ml) to prepare a sample for measurement; separately, add pepsin from pig gastric mucosa to 0.1N hydrochloric acid aqueous solution to dissolve it at 1200Units/ml, and filter it to prepare a pepsin solution; add 0.1g of the sample to be measured to 40mL of the pepsin solution, stir it in an incubator at 37℃ with a stirrer, and calculate the time until the cross-linked gelatin (insoluble matter) disappears as the pepsin digestion time.