KR20100132878A - Anti-adhesion agent comprising epoxide-crosslinked hyaluronic acid derivative hydrogel and process for producing the same - Google Patents

Abstract

PURPOSE: An anti-adhesion agent containing hyaluronic acid epoxide derivative hydrogel and a method for manufacturing the same are provided to prevent adhesion between tissues without side effects. CONSTITUTION: An anti-adhesion agent contains hyaluronic acid epoxide derivative hydrogen. The particle size of the hydrogel is 100-300 um. The viscosity and crosslinking density of the hydrogel are 100,000 cP (0.02 Hz)-6,000,000 cP (0.02 Hz) and 0.2 mol%-100 mol%, respectively. The hydrogel is prepared by reacting hyaluronic acid and epoxide crosslinking agent in NaOH solution or crossliking 0.1-200 weight parts of crosslinking agent based on 100 weight parts of repeat unit of the hyaluronic acid.

Description

Anti-adhesion agent comprising hyaluronic acid epoxide derivative hydrogel and a method for preparing the same {ANTI-ADHESION AGENT COMPRISING EPOXIDE-CROSSLINKED HYALURONIC ACID DERIVATIVE HYDROGEL AND PROCESS FOR PRODUCING THE SAME}

The present invention relates to an anti-adhesion agent comprising a hyaluronic acid epoxide derivative hydrogel and a preparation method thereof. More specifically, the present invention relates to an anti-adhesion agent including a hyaluronic acid epoxide derivative hydrogel used for the purpose of preventing adhesion between tissues after surgery, and a method for preparing the same.

After surgery, or when tissue damage occurs due to inflammation, foreign bodies, bleeding, infection, wounds, friction, etc., blood leaks during the healing of wounds and coagulates, resulting in abnormal adhesion to surrounding tissues. Occurs. If cells invade these adhesion sites, stronger adhesion occurs.

In general, adhesion occurs at a frequency of about 60% to 95% after open surgery, and may cause pain, intestinal obstruction, infertility, and the like, and reoperation is required to remove adhesion due to organ or tissue dysfunction. In particular, tissue adhesion is a major cause of intestinal obstruction (80% to 90%) and causes long-term sequelae and pelvic pain after gynecology ( Eur. J. Surg. 1997, Supp1 577 : 32-39).

Methods to prevent such adhesion include minimizing wounds during surgery, using anti-inflammatory agents ( Sepsis 1999, 3 : 317-25), increasing tPA activity ( British J. Surg . 2001, 88 : 286), and physical barriers. (barrier) has been developed and used ( Eur. J. Surg. 1997, Supp 1 577: 40). Among them, a physical barrier, that is, a material suitable as an anti-adhesion agent, acts as a barrier only during the healing of wounds in the body, and after that, it must be decomposed, and there is no toxicity of the material itself. It should be harmless to.

So far, various materials have been studied that can provide a physical barrier. Water-soluble polymers such as dextran, hyaluronic acid (HA), carboxymethylcellulose (CMC), and polyethylene glycol (poly (ethylene glycol) ), PEG), Pluronic, etc., poly (lactic acid), PLA), polyglycolic acid (PGA), polylactic acid-glycolic acid copolymers poly (lactic-co-glycolic acid), PLGA).

Hyaluronic acid is a non-sulfated glycosaminoglycan that is widely distributed throughout connective, epithelial and neural tissues. Hyaluronic acid is composed of D-glucuronic acid and D-N-acetylglucosamine, and is a disaccharide polymer in which β-1,4 and β-1,3 glucosidic bonds are alternately bonded.

Seprafilm ^TM (Genzyme Corp. Cambridge, MA), which is cross-linked with Interceed ^TM (Johnson & Johnson Medical, Inc. Arlinton, TX, USA), HA, and CMC using biodegradable Oxidized Regenerated Cellulose (ORC) materials , USA), Guardix ^TM (developed by Korea Biolane Co., Ltd., Hanmi Pharm.), Which is simply a mixture of HA and CMC, has been commercialized and widely used.

The commercialized Interceed ^TM has low biocompatibility because the main raw material ORC is a non-living material, and it has a very large pore fabric, and it is easy to permeate cells and blood proteins, resulting in low efficiency as a separation membrane. ( J. of Surgical Research 1997, 68 : 126-132).

Pluronic F127 anti-adhesion agents have the advantage of ease of use, which is a sol form at room temperature and gel form at body temperature, but has the disadvantage that it should be used only on dry, bleed-free surfaces. In addition, the anti-adhesion agent made of PLA or PGA, which is a biodegradable synthetic polymer, has a long biodegradation period of 4 weeks or more and a high mechanical strength. However, since it is a hydrophobic material, it does not adhere well to the surface of the biological tissue due to its low water absorption. It is known that hydrolysis may cause acidic decomposition products to be released, thereby causing an inflammatory reaction that causes adhesion.

In addition, Seprafilm ^TM combines HA and CMC in a ratio of 2: 1 and crosslinking activator 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1-ethyl-3- (3-dimethylaminopropyl) Only carbodiimide hydrochloride (EDC) is reacted to form N-acylurea pendants (US Pat. Nos. 5,017,229, 5,527,893 and 5,760,200).

In the method of preparing a derivative of the patent, a derivative is prepared through a process of drying a HA and CMC mixture to which an EDC, which is a crosslinking activator, is dried, and removing unreacted substances and impurities from the prepared film. The method is limited in making derivatization at the same time as the film is made to make a uniform and transparent film.

In addition, since the film is derivatized as a whole, in vivo degradation characteristics, the derivative portion composed of HA is easily decomposed (about 66% degradation), and the CMC derivative portion (about 33%) is more than necessary. It remains a long time.

In addition, fine pores are formed in the process of removing impurities from the manufactured film, and are not easily handled because they are easily broken in a dry state. In addition, the film curls when contacted with moisture, which results in different results depending on the application site, and does not prove a definite effect.

On the other hand, hyaluronic acid (HA), which is highly biocompatible, has been shown to have an effect of smoothing the intestinal surface and reducing the formation of intraperitoneal adhesions after surgery. Guardix ^TM , first commercialized in Korea as a anti-adhesion agent mixed with CMC and HA, is a gel type that is different from the conventional anti-adhesion agent, and can be applied to various surgical sites and is easily absorbed in vivo. Since HA is used as it is without deformation, the effect of preventing adhesion decreases.

International patent application WO 1997/07833 discloses new biomaterials essentially containing esterified derivatives of hyaluronic acid or self-crosslinked derivatives of hyaluronic acid, their use in the surgical field, in particular for preventing postoperative adhesions. It starts.

However, the biomaterial according to the international patent application is a cross-linked carboxyl group and a hydroxyl group of hyaluronic acid itself, that is, auto-crosslinking, which is crosslinked using an epoxide crosslinking agent. Lonic acid epoxide derivatives are distinct substances that differ in their chemical structure.

In addition, compared with the biomaterial according to WO 1997/07833, the hyaluronic acid epoxide derivative hydrogel of the present invention, by using an epoxide crosslinking agent, it is easy to control the decomposition and absorption rate in vivo, Its adhesion to biological tissues is excellent due to its water absorption and excellent adhesion.

Moreover, the hyaluronic acid epoxide derivative hydrogel of the present invention is convenient for use because of its excellent injection performance when swollen with saline or the like.

In the prior patent application of the inventors (Patent Application No .: 10-2007-0076258), the film was prepared by the surface cross-linking of HA and CMC to develop an anti-adhesion agent having improved flexibility and physical strength, but relatively limited to the application area And the epoxide conjugates for the complex of HA and CMC remain longer than necessary for the CMC derivative moiety.

Therefore, the present inventors have newly studied an adhesion agent which has high tissue adhesion and anti-adhesion ability, easy-to-use hydrogel form, completely decomposed and absorbed in vivo, and non-toxic in order to overcome the above-mentioned problems of the prior art. Overcoming the problems of the prior art has led to the development of new and effective anti-adhesion agents of the present invention.

It is a basic object of the present invention to provide an anti-adhesion agent comprising a hyaluronic acid epoxide derivative hydrogel.

Unlike conventional hyaluronic acid anti-adhesion agents, the anti-adhesion agent of the present invention is characterized by excellent adhesion to biological tissue, complete degradation and absorption in vivo after a period of time, and excellent injection ability.

In addition, it is characterized by providing an anti-adhesion agent of a HA derivative in the form of a hydrogel, without using the CMC derivative portion that has been pointed out as a problem in the prior art.

The anti-adhesion agent is i) reacting the hyaluronic acid dissolved in the basic aqueous solution with the epoxide crosslinking agent to prepare a hyaluronic acid epoxide derivative; ii) pulverizing the hyaluronic acid epoxide derivative may be prepared through a method for preparing an anti-adhesion comprising a hyaluronic acid epoxide derivative hydrogel.

The basic object of the present invention described above can be achieved by providing an anti-adhesion agent comprising a hyaluronic acid epoxide derivative hydrogel.

The hyaluronic acid epoxide derivative of the present invention is obtained by crosslinking hyaluronic acid with an epoxide crosslinking agent having two or more epoxide groups. The crosslinking density of the hyaluronic acid epoxide derivative of the present invention is 0.2 mol% to 100 mol%.

 When the molecular weight of the hyaluronic acid epoxide derivative of the present invention is calculated from the crosslinking density, it is arithmetically distributed in the range of several hundred million Da to several hundred billion Da.

Since the anti-adhesion agent is preferably in the form of a hydrogel having a micrometer size, more preferably 100 μm to 300 μm size, it is possible to spray or inject at the site where adhesion can occur. Therefore, it is simpler to use than the film anti-adhesion agent.

The complex viscosity of the hyaluronic acid epoxide derivative hydrogel of the present invention is 100,000 cP (0.02 Hz) to 6,000,000 cP (0.02 Hz).

In addition, the hyaluronic acid epoxide derivative hydrogel has a high water absorption capacity (Fig. 2) has a large swelling degree. In the gel state, the swelling degree is 300% to 1,500%, and in the case of the hyaluronic acid epoxide derivative powder, the swelling degree is 300% to 75,000%.

Moreover, the hyaluronic acid epoxide derivative hydrogel has a high viscoelasticity (FIG. 1), which is excellent in adhesion to biological tissues and has adequate stability to hyaluronic acid degrading enzymes, thereby forming a physical barrier in the affected area for a predetermined time. And then completely decomposed and absorbed into the living body (FIGS. 5 to 8).

The anti-adhesion agent of the present invention comprises the steps of: i) reacting hyaluronic acid dissolved in a basic aqueous solution with an epoxide crosslinking agent to prepare a hyaluronic acid epoxide derivative; ii) pulverizing the hyaluronic acid epoxide derivative may be prepared through a method for preparing an anti-adhesion comprising a hyaluronic acid epoxide derivative hydrogel.

Hyaluronic Acid Epoxide Derivatives According to the present invention, a hyaluronic acid epoxide derivative prepared in the form of particles by pulverizing a reactant obtained by reacting hyaluronic acid (HA) with an epoxide crosslinking agent containing two or more epoxy groups. Prepare a hydrogel.

The structure of hyaluronic acid is represented by following formula (1).

Hyaluronic acid (HA) in the present invention includes hyaluronic acid and salts thereof, the salt is sodium hyaluronate, potassium hyaluronic acid, calcium hyaluronic acid, magnesium hyaluronate, zinc hyaluronate, cobalt hyaluronic acid or tetrabutylammonium hyaluronic acid At least one of the.

The anti-adhesion preparation method including the hyaluronic acid epoxide derivative hydrogel of the present invention may be represented by the following Scheme 1.

As the basic aqueous solution used in step i) of the method of the present invention, NaOH aqueous solution or the like may be used. In addition, the reaction temperature of step i) is 4 ℃ to 80 ℃, the reaction time is 12 hours to 48 hours, and the reaction pressure is preferably atmospheric pressure.

The epoxide crosslinking agent of step i) 1,4-butanediol diglycidyl ether (BDDE), ethylene glycol diglycidyl ether (ethylene glycol diglycidyl ether, EGDGE), 1,6 Hexanediol diglycidyl ether (1,6-hexanediol diglycidyl ether), propylene glycol diglycidyl ether, poly (propylene glycol) diglycidyl ether, Poly (tetramethylene glycol) diglycidyl ether, neopentyl glycol diglycidyl ether, polyglycerol polyglycidyl ether, diglycerol Diglycerol polyglycidyl ether, glycerol polyglycidyl ether, trimethylpropane polyglycidyl ether, 1,2- (non (2,3-epoxypropoxy) ethylene (1,2- (bis (2,3-epoxypropoxy) ethylene), pentaerythritol polyglycidyl ether or sorbitol polyglycidyl ether Can be selected from.

The weight ratio of the hyaluronic acid and the epoxide crosslinking agent in step i) is 0.1 to 200 parts by weight, more preferably 0.1 to 50 parts by weight of the epoxide crosslinking agent relative to 100 parts by weight of the repeating unit of the hyaluronic acid. You can.

The hyaluronic acid epoxide derivative prepared in step i) is washed with physiological saline, etc., and then ground to prepare a hydrogel. The prepared hydrogel can be used by sterilization under high temperature and high pressure conditions. In particular, the size and shape of the hyaluronic acid epoxide derivative is not particularly limited, it may be varied in size and shape according to the medically applied range.

In addition to the washing method, the hyaluronic acid epoxide derivative prepared in step i) is a method known in the art, such as distillation (at atmospheric pressure or reduced pressure), recrystallization, column chromatography, ion exchange chromatography, gel chromatography, It may be separated and / or purified by affinity chromatography, thin layer chromatography, phase separation, solvent extraction or dialysis.

The composite viscosity of the hydrogel prepared according to the hyaluronic acid epoxide derivative hydrogel production method of the present invention is 100,000 cP (0.02 Hz) to 6,000,000 cP (0.02 Hz), the crosslinking density is 0.2 mol% to 100 mol%, and the swelling degree is 300% to 1,500%. In addition, when the hyaluronic acid epoxide derivative is in a powder state, the degree of swelling reaches 75,000%.

In addition, the size of the hydrogel pulverized in step ii) is preferably 100 μm to 300 μm, but may be used by grinding to another size as necessary.

The micrometer-sized hyaluronic acid epoxide derivative hydrogel according to the present invention has a high water absorption capacity. In addition, when applied to a living body, it initially maintains a stable form to prevent adhesion between tissues, and is completely decomposed and absorbed over time. Moreover, not only the adhesion to biological tissues is very good, but also the injection performance is excellent and convenient to use.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following description of the embodiments is only intended to specifically describe the specific embodiments of the present invention, it is not intended to limit or limit the scope of the present invention to the contents described therein.

Example  1 hyaluronic acid epoxide derivative Hydrogel  Produce

10 ml of hyaluronic acid solution in which 1,000 mg of hyaluronic acid was dissolved in 0.25 N NaOH solution was prepared in four reactors, respectively. To the hyaluronic acid solution was added 25 mg, 50 mg, 100 mg and 200 mg of butanediol diglycidyl ether (BDDE), respectively. After reacting for 24 hours at room temperature, washed with physiological saline to remove the unreacted. The washed product was pulverized to adjust particle size and concentration, and then sterilized at 121 ° C. for 15 minutes to prepare a hyaluronic acid epoxide derivative hydrogel.

Example  2. Hyaluronic Acid Epoxide Derivatives Hydrogel Rheological  characteristic

Rheometer is used to compare the rheological properties of the hyaluronic acid epoxide derivative hydrogel prepared in Example 1, the hyaluronic acid-containing anti-adhesion agent approved by the Korea Food and Drug Administration, and the general hyaluronic acid injection Complex Viscosity and tan δ were measured in the frequency range of 0.02 Hz to 1 Hz.

1 is a comparison of the physical properties of the hyaluronic acid epoxide derivative hydrogel of the present invention and commercially available hyaluronic acid-containing anti-adhesion agent, and a general hyaluronic acid injection, the hyaluronic acid epoxide derivative hydrogel of the present invention is commercialized as an anti-adhesion agent The composite viscosity value is much better than hyaluronic acid-containing products and general hyaluronic acid injections. In addition, looking at the tan δ value, it can be seen that the network structure having structurally high stability is formed in the hyaluronic acid epoxide derivative hydrogel of the present invention.

Example  3. Hyaluronic Acid Epoxide Derivatives Hydrogel Absorption

To 0.6 ml of the hyaluronic acid epoxide derivative hydrogel prepared in Example 1, 10 ml of physiological saline was added and stirred vigorously for 5 minutes. After standing at 37 ° C. for 72 hours, the volume of the solution absorbed in the hyaluronic acid epoxide derivative hydrogel was confirmed.

2 is a result of the absorption capacity of the hyaluronic acid epoxide derivative hydrogel, the hyaluronic acid epoxide derivative hydrogel of the present invention showed the ability to absorb water up to 6 times its volume while maintaining the form of the particles.

Example  4. Hyaluronic Acid Epoxide Derivatives Gel Adhesion

In order to measure the tack value of the hyaluronic acid epoxide derivative hydrogel prepared in Example 1 and a commercial hyaluronic acid-containing anti-adhesion agent and a general hyaluronic acid injection approved by the Korea Food and Drug Administration, the plate-plate of the rheometer After loading (loading), the rotation was performed at a shear rate of 10 ^-1 for 0.1 seconds (Normal Force) value was measured when the upper plate was dropped at a rate of 0.1 mm / s.

Figure 3 is a comparison of the talc value of the hyaluronic acid epoxide derivative hydrogel of the present invention, the hyaluronic acid epoxide derivative hydrogel is a hyaluronic acid containing hyaluronic acid containing licensed from the Korea Food and Drug Administration Adhesion inhibitors and talc values were higher than general hyaluronic acid injections. That is, the hyaluronic acid epoxide derivative hydrogel of the present invention showed a higher adhesion than the comparative objects.

Example  5. Hyaluronic Acid Epoxide Derivatives from Experimental Animal Models Hydrogel  Anti-adhesion Efficacy

Five Sprague Dawley rats were divided into control and experimental groups, and the abdomen was opened, and the cecum and abdominal wall were scratched. Afterwards, the negative control group without any anti-adhesion agent, the imported anti-adhesion film (S film of G company), the gel (G gel of H company) licensed as an anti-adhesion agent from Korea Food and Drug Administration, and a general hyaluronic acid injection ( The change of adhesion in the experimental group using the hyaluronic acid epoxide derivative hydrogel prepared in Example 3 and the positive control group of Group 3 using L company H product) was compared and observed.

The efficacy test is described in more detail as follows. 230 g to 250 g of Sprague Dawley rats were divided into 5 groups of control and experimental groups, respectively, followed by anesthesia by intramuscular injection of dilutions of ketamine 5 mg / 100 g and ampicillin 10 mg over a 2 week period. The abdomen was dissected. Thereafter, the caecum was injured to a size of 1 cm to 2 cm, and the opposing peritoneal membrane was damaged to the same size, and then the positive control group and the experimental group were sufficiently covered with a film or gel between the cecum and the abdominal wall. And two portions 1 cm away from the injury site were fixed with sutures (FIG. 4). Two weeks after the operation, the rats were sacrificed to evaluate the degree of adhesion, adhesion strength, and adhesion area.

In the evaluation of adhesion, the degree of adhesion was evaluated based on the following criteria.

-Without adhesion: 0

One thin film of coalescence: 1

Two or more thin film-type coalescences: 2

-Concentrated thick coalescing on spots: 3

-Plated concentrated thick coalescence: 4

-Very thick adhesions in which blood vessels are formed or one or more plate-like thick adhesions: 5

In addition, in the evaluation of adhesion, adhesion intensity was evaluated based on the following criteria.

-Without adhesion: 0

-Adhesions that are film-like and fall with very weak force: 1

Adhesions requiring moderate force: 2

-Coalescing that requires significant pressure: 3

-Adhesions are so strong that they are difficult to remove or require very high pressure: 4

As a result of the evaluation of adhesion after the experiment, as shown in FIGS. 5 to 8, the hyaluronic acid epoxide derivative hydrogel of the present invention showed that adhesion was formed between adjacent tissues while the wound at the injury site was healed compared to the negative control group. It clearly shows that it is blocking. In addition, as shown in Figure 5, the experimental group using the hyaluronic acid epoxide derivative hydrogel of the present invention compared to the negative control group showed a small adhesion degree to the tissue separated from the surrounding tissue.

In particular, as shown in Figure 6, the degree of adhesion of the experimental group using the hyaluronic acid epoxide derivative hydrogel of the present invention is 0.6 ± 1.34, not only the negative control group of 4.7 ± 0.35 but also the permission of the Korea Food and Drug Administration of 3.3 ± 0.75 The degree of adhesion was higher than that of the positive control group such as the anti-adhesion agent received (G gel of H company), the general hyaluronic acid injection of 2.2 ± 0.95 (H product of L company), and the imported anti-adhesion film of 2.2 ± 1.79 (G film S film). Low.

The adhesion strength of the experimental group using the hyaluronic acid epoxide derivative hydrogel of the present invention was measured to 0.3 ± 0.76, as shown in Figure 7, the negative control group of 3.8 ± 0.16, as well as the permission of the Korea Food and Drug Administration of 2.4 ± 0.31 Very small values compared to the positive control group of the anti-adhesion agent (G gel of H company), the general hyaluronic acid injection of 1.3 ± 0.41 (H product of L company), and the imported anti-adhesion film of 1.7 ± 1.17 (S film of G company). Showed.

In addition, as shown in Figure 8, the adhesion area of the experimental group using the hyaluronic acid epoxide derivative hydrogel of the present invention was measured to 0.05 ± 0.11 cm ² which is 2.48 ± 0.5 cm ² negative control group and 0.25 ± 0.15 cm ² Adhesion inhibitors approved by the Korea Food and Drug Administration (G gel of H), general hyaluronic acid injections of 0.1 ± 0.11 cm ² (H product of L company), and imported adhesion inhibitor films of 0.4 ± 0.41 cm ² (G S film) ), It showed a smaller value.

As a result of the anti-adhesion efficacy test using an animal, the hyaluronic acid epoxide derivative hydrogel of the present invention showed the lowest value in comparison with the positive control group as well as the negative control group in terms of adhesion strength, adhesion degree as well as the adhesion area.

Example  6. Hyaluronic Acid Epoxide  derivative Hydrogel Injection  Measure

In order to confirm the injection ability of the hyaluronic acid epoxide derivative hydrogel prepared in Example 1, the contents were filled in a 1 ml syringe into which a catheter was inserted, using an injection force measuring device, and then at a speed of 12.5 mm / min. Injection force was measured by pushing the syringe rod. The measured value was a low value of 11.368 N.

Example  7. Hyaluronic Acid Epoxide  derivative Hydrogel  Safety check

Non-clinical toxicity testing was performed by a GLP certification institution for the safety confirmation test of hyaluronic acid epoxide derivative hydrogel (Table 1).

Nonclinical Toxicity Test Results of Hyaluronic Acid Epoxide Derivative Hydrogel Test Items Test Methods Test standard Test result Cytotoxicity test Performed in accordance with ISO 10993-5 and USP24-NF19 Biological Tests Cell growth deterioration, lysis No cytotoxicity was observed Irritation test Performed in accordance with ISO 10993-10 General symptoms, weight measurement, skin reaction judgment, evaluation of skin reaction results Corresponds to non-irritating substances Acute Systemic Toxicity Test Performed in accordance with ISO 10993-11 and 'Common Standards on Biological Safety of Medical Devices' Notice No. 2006-32 General clinical symptoms, weight measurement, autopsy and pathological examination, behavioral abnormalities No toxicity. Harmless. A sensitization test Performed in accordance with ISO 10993-10 General symptoms, weight measurement, skin reaction judgment, evaluation of skin reaction results No sensitivity Chromosome Aberration Test Common Standards on Biological Safety of Medical Devices, Korea Food and Drug Administration Notification No. 2006-32, ISO10993-3, 'Toxicological Standards for Drugs, etc.' Food and Drug Administration Notification No. 2005-60, OECD Toxicity Test Guidelines ( No. 473), performed by Cluture of Animal Cells-4th ed. Increasing frequency of abnormal middle phase and abnormal number Does not mutate Revert mutation test Common Criteria Standard for Biological Safety of Medical Devices, Korea Food and Drug Administration Notification No. 2006-32, ISO 10993-3, 'Toxicological Standards for Drugs,' Food and Drug Administration Notification No. 2005-60, OECD Test Guideline ( No. 471), carried out by the OECD Test Guideline (No. 472). Inhibition of growth, increase in colony count Does not mutate Micronuclear test 'Common Criteria Standard for Biological Safety of Medical Devices' Korean Food and Drug Administration Notification No. 2006-32, ISO 10993-3, 'Toxicological Standards for Drugs,' Food and Drug Administration Notification No. 2005-60, OECD Toxicity Test Guidelines Performed in accordance with (No. 474) General clinical signs, body weight, micronucleus count Test substance does not cause micronucleus Pyrogenic test Performed according to USP29-NF24 <151> Pyrogen Test Clinical symptoms, behavioral abnormalities, deaths, control temperature measurement, peak temperature measurement Pyrogenic voice Transplantation / subchronicity test Performed in accordance with ISO 10993-6, ISO 10993-11, Common Criteria Standard for Biological Safety of Medical Devices (KFDA Notification 2006-32) Clinical symptoms, weight measurement, feed and drinking intake, urine test, ophthalmological test, general hematological and hemochemical test, pathological test Systemic toxicity was not observed.
No irritation. Hemolytic test Performed in accordance with ISO 10993-4 Absorbance Measurement, Hemolytic Rate Measurement No hemolytic

As a result of the non-clinical toxicity test, it was confirmed that the hyaluronic acid epoxide derivative hydrogel of the present invention was a safe substance.

FIG. 1a compares the combined viscosity of a hyaluronic acid epoxide derivative hydrogel of the present invention, a commercially available hyaluronic acid-containing anti-adhesion agent, and a general hyaluronic acid injection, and FIG. 1b compares tanδ thereof.

Figure 2 shows the water absorption of the hyaluronic acid epoxide derivative hydrogel of the present invention.

Figure 3 is a hyaluronic acid epoxide derivative hydrogel of the present invention shows the talc value of the hyaluronic acid-containing anti-adhesion agent, and the general hyaluronic acid injections sold and approved by the Korea Food and Drug Administration.

Figure 4 shows the anti-adhesion efficacy test method by a rat animal model.

Figure 5 is a rat animal model, the negative control group without using the anti-adhesion agent, the experimental group using the hyaluronic acid epoxide derivative hydrogel of the present invention, and the imported anti-adhesion film (S film of G company), the adhesion from the Korea Food and Drug Administration, Photographs of three groups of positive control group using gel (G gel of H company) licensed as an inhibitor and general hyaluronic acid injection (H product of L company) are shown.

FIG. 6 shows the degree of adhesion in the negative control group, the experimental group, and the positive control group (group 3) of FIG. 5.

7 shows adhesion strengths in the negative control group, the experimental group, and the positive control group (group 3) of FIG. 5.

FIG. 8 shows adhesion areas in the negative control group, the experimental group, and the positive control group (group 3) of FIG. 5.

Claims (7)

An anti-adhesion agent comprising a hyaluronic acid epoxide derivative hydrogel. The anti-adhesion agent according to claim 1, wherein the hyaluronic acid epoxide derivative hydrogel is in the form of particles having a size of 100 μm to 300 μm. The anti-adhesion agent according to claim 1, wherein the complex viscosity of the hyaluronic acid epoxide derivative hydrogel is 100,000 cP (0.02 Hz) to 6,000,000 cP (0.02 Hz). The anti-adhesion agent according to claim 1, wherein the hyaluronic acid epoxide derivative hydrogel has a crosslinking density of 0.2 mol% to 100 mol%. The anti-adhesion agent according to claim 1, wherein the hyaluronic acid epoxide derivative hydrogel is prepared by reacting hyaluronic acid with an epoxide crosslinking agent in an aqueous NaOH solution. The method of claim 1, wherein the hyaluronic acid epoxide derivative hydrogel is 1,4-butane diol diglycidyl ether, ethylene glycol diglycidyl ether, 1,6-hexanediol diglycidyl ether, propylene glycol di Glycidyl ether, polypropylene glycol diglycidyl ether, polytetramethylene glycol diglycidyl ether, neopentyl glycol diglycidyl ether, polyglycerol polyglycidyl ether, diglycerol polyglycidyl ether, Glycerol polyglycidyl ether, trimethylpropane polyglycidyl ether, 1,2- (bis (2,3-epoxypropoxy) ethylene, pentaerythritol polyglycidyl ether and sorbitol polyglycidyl ether Anti-adhesion agent, characterized in that prepared by reacting the selected epoxide crosslinking agent and hyaluronic acid. According to claim 1, wherein the hyaluronic acid epoxide derivative hydrogel cross-linking reaction of the hyaluronic acid and epoxide crosslinking agent in the ratio of 0.1 to 200 parts by weight of the epoxide crosslinking agent relative to 100 parts by weight of the repeating unit of the hyaluronic acid Adhesion inhibitors, characterized in that prepared by.

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