KR20090012439A - Hyaluronic acid and carboxymethylcellulose complex derivative film and gel having anti-adhesion property and producing method thereof - Google Patents

Abstract

A hyaluronic acid and a carboxy methyl cellulose composite derivative film is provided to have trachea reducing adhesion property, excellent biodegradation property and property absorbed by being completely disassembled after preventing adhesion phenomenon. An N-acylation urea pendant form and an auto-cross-linking form are mixed in hyaluronic acid and carboxy methyl cellulose composite derivative film. The N-acylation urea pendant form and an auto-cross-linking form are made by reacting crosslink activity agent and cross linkage activity supplement at hyaluronic acid and carboxy methyl cellulose complex film surface. The derivative film with the N-acylation urea pendant form is formed by using the crosslink activity agent 0.01 ~ 500 parts by weight and the cross linkage activity supplement 0.01 ~ 200 parts by weight based on compound of the carboxy methyl cellulose and hyaluronic acid 100 parts by weight. The derivative film with the auto-cross-linking form is formed by using the crosslink activity agent 0.01 ~ 500 parts by weight and the cross linkage activity supplement 40 ~ 1000 parts by weight.

Description

Hyaluronic acid and carboxymethylcellulose complex derivative film and gel and anti-adhesion property and producing method

The present invention relates to a hyaluronic acid and carboxymethyl cellulose composite derivative film and gel and a method for preparing the same, and more particularly, to derivatization of hyaluronic acid and carboxymethyl cellulose complex of various compositions with excellent biocompatibility. Hyaluronic acid prevents adhesions between organs and surrounding organs after surgical operation by adjusting the morphology and derivatization method, and prevents adhesion to organs within 4 weeks, greatly reducing the possibility of side effects. And it relates to a carboxymethyl cellulose composite derivative film and gel and a method for producing the same.

If tissue damage occurs after surgery or inflammation, foreign bodies, bleeding, infection, wounds, friction, etc., blood leaks during the healing of wounds and coagulates, resulting in adhesions that cause abnormal junctions with surrounding tissues. do. When cells penetrate here, 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 reoperation to remove adhesion due to organ or tissue dysfunction. In particular, tissue adhesion is the main cause of intestinal obstruction (80-90%), and long-term sequelae and pelvic pain after gynecological procedures (Eur. J. Surg. 1997, Supp1577, 32-39).

As a method of preventing such adhesion, a method of minimizing wounds during surgery or using an anti-inflammatory agent and using a physical barrier has been developed and used. Among them, the material used as a physical barrier acts as a barrier only during the period of wound healing in the body, and after that, it must be decomposed, and there is no toxicity of the material itself. It should be harmless to human body.

Various materials have been studied in the past, water-soluble polymers such as dextran, sodium hyaluronate (HA), sodium carboxymethylcellulose (CMC), polyethyleneglycol, and pluronic Polylactic acid (PLA), polyglycolic acid (PGA), and polylactic-co-glycolic acid (PLGA), which are non-aqueous synthetic polymers. Interceed ^TM (Johnson & Johnson Medical, Inc. Arlinto, TX) and Seprafilm ^TM (Genzyme Cop. Camridge, MA), which are used as cellulose-oxidized regenerated cellulose (ORC) materials, have been commercialized and used.

First commercialized, Interceed ^TM is a non-living material that has low biocompatibility, and has a very large pore fabric that allows easy penetration of cells and blood proteins. (J. of Surgical Research, 68, 126-132, 1997). Pluronic F127 has the advantage of ease of use, which is sol form at room temperature and gel form at body temperature, but has the disadvantage of limited use only on dry, bleed-free surfaces. In addition, the anti-adhesion agent made of biodegradable synthetic polymer PLA and PGA has a high biomechanical strength with a long biodegradation period of 4 weeks or more. In addition, it may hydrolyze in vivo to release acidic degradation products, which may cause some inflammatory reactions that cause adhesion.

On the other hand, hyaluronic acid has been shown to reduce intraperitoneal adhesion formation in animal studies by smoothing the intestinal surface and preventing postoperative adhesion formation (SIHOLZMAN, MD, 1994). However, it could not be rapidly absorbed in vivo and exhibit the desired anti-adhesion effect.

     In order to solve this problem, there is a known technique for derivatization of a lyophilized hyaluronic acid using a crosslinking agent (US Pat. No. 6,630,167). As a result, there was a problem that there was a limit in the area of coalition.

In addition, Seprafilm ^™ is a 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1-ethyl-3- (3-dimethylaminopropyl) which is a crosslinking activator with a 2: 1 blend of hyaluronic acid and carboxymethylcellulose. The reaction is carried out using only carbodiimide hydrochloride (EDC) to form N-acylurea pendant (US Patent Nos. 5,017,229, 5,527,893 and 5,760,200). In the above patented derivatization production method, a hyaluronic acid and a carboxymethyl cellulose mixture to which an EDC, which is a crosslinking activator, are dried to prepare a film, and a non-reactant and impurities are removed from the manufactured film. The method is derivatized at the same time while the film is manufactured, there is a limitation in making a uniform and transparent film, and also the film is derivatized in the same way as a whole, and when the decomposition properties in vivo, the derivative consisting of hyaluronic acid initially The portion is easily degraded (about 66% degradation) and the carboxymethylcellulose derivative portion (about 33%) remains in shape longer than necessary. In addition, fine pores are formed in the process of removing impurities from the manufactured film, and thus are not easily handled because they are brittle in a dry state. The film curls when contacted with moisture, and the results are different depending on the experimental model, which does not prove a definite effect.

The present invention is to solve the problems of the prior art while maintaining a stable form at the beginning while showing flexibility, to provide a hyaluronic acid and carboxymethyl cellulose composite derivative film and gel which is completely decomposed and absorbed after preventing the adhesion phenomenon The purpose is.

The above object of the present invention is the hyaluronic acid and carboxymethyl in which the N-acylurea pendant form and the auto-crosslinking form are derivatized by reacting a crosslinking activator and a crosslinking activator on the surface of the hyaluronic acid and the carboxymethylcellulose composite film. By forming a cellulose composite derivative film.

In addition, the above object of the present invention can be achieved by reacting a hyaluronic acid and a carboxymethyl cellulose complex with a crosslinking agent comprising two or more epoxy functional groups to form a derivatized hyaluronic acid and a carboxymethyl cellulose complex derivative gel.

In addition, the present invention is to provide a method for producing a hyaluronic acid and carboxymethyl cellulose composite derivative film and gel, which does not break the derivative film or gel, is dried upon contact with water or the result is constant according to the experimental model.

The object of the present invention is the step of dissolving hyaluronic acid and carboxymethyl cellulose in distilled water; Stirring the dissolved hyaluronic acid and carboxymethylcellulose to form a film by drying; Adding a crosslinking activator and a crosslinking activity aid to the aqueous organic solvent and then mixing the crosslinking reaction on the surface of the film; Purifying the surface crosslinked film using a washing solvent; Drying the purified film; And sterilizing the dried film by gamma irradiation. It can be achieved by providing a hyaluronic acid and a carboxymethylcellulose complex derivative film manufacturing method comprising a.

In addition, the object of the present invention is the step of dissolving hyaluronic acid and carboxymethyl cellulose in an aqueous alkali solution; Reacting the hyaluronic acid with a carboxymethyl cellulose aqueous alkali solution by adding a crosslinking agent; Washing the crosslinked hyaluronic acid and carboxymethyl cellulose with distilled water; Swelling the washed reactant in physiological saline to form a gel; It can be achieved by providing a method for producing a hyaluronic acid and carboxymethyl cellulose complex derivative gel comprising a; and sterilizing the gel at a high temperature and high pressure.

The present invention relates to a hyaluronic acid and carboxymethyl cellulose composite derivative film and gel and a method for preparing the same, and more particularly, to derivatization of hyaluronic acid and carboxymethyl cellulose complex of various compositions with excellent biocompatibility. Hyaluronic acid prevents adhesions between organs and surrounding organs after surgical operation by adjusting the morphology and derivatization method, and prevents adhesion to organs within 4 weeks, greatly reducing the possibility of side effects. And it relates to a carboxymethyl cellulose composite derivative film and gel and a method for producing the same.

Here, the hyaluronic acid and carboxymethyl cellulose complex derivative film and gel of the present invention has the advantage of harmless to the human body melting itself after forming a physical wall for a certain period of time, so as to prevent long-term adhesion that is an aftereffect of surgery, It is an anti-adhesion agent that combines anti-adhesion performance and ease of use with improved adhesion, flexibility and physical strength to tissues.

Specifically, the present invention, in the hyaluronic acid and carboxymethyl cellulose composite derivative film, N-acylurea pendant form derivatized by reacting a crosslinking activator and a crosslinking activity adjuvant to the surface of the hyaluronic acid and carboxymethyl cellulose composite film Provided are a hyaluronic acid and a carboxymethylcellulose composite derivative film mixed with auto-crosslinking forms.

At this time, the derivative film of the N-acylurea pendant form is predominant using 0.01 to 500 parts by weight of the crosslinking activator and 0.01 to 200 parts by weight of the crosslinking activator with respect to 100 parts by weight of the repeating unit of the hyaluronic acid and the carboxymethyl cellulose complex. Or 0.01 to 500 parts by weight of the crosslinking activator and 40 to 1000 parts by weight of the crosslinking activator to form a derivative film having an auto-crosslinked form.

Here, the hyaluronic acid and carboxymethyl cellulose composite derivative film according to the present invention is dried by dissolving the composite aqueous solution in which the hyaluronic acid and carboxymethyl cellulose mixed in an aqueous solution to form a film, preventing adhesion and excellent adhesion and flexibility It is a derivative film having a crosslinking reaction on the surface of the film by mixing a crosslinking activator and a crosslinking activity aid so as to be used as a biofilm, and is formed to be degraded, metabolized, and released in vivo within 4 weeks after preventing adhesion.

Since the fine pores are not formed and the film is crosslinked on a uniform and transparent film surface, the film is flexible and physically supported, and the usability is greatly improved.

And the hyaluronic acid includes hyaluronic acid and salts thereof, wherein the salt is at least one of sodium hyaluronate, potassium hyaluronic acid, calcium hyaluronic acid, magnesium hyaluronic acid, zinc hyaluronic acid, cobalt hyaluronic acid, or tetrabutylammonium hyaluronic acid. It includes one or more.

In addition, the carboxymethyl cellulose has a degree of substitution of 0.2 to 1.5, characterized in that the sodium carboxymethyl cellulose having a molecular weight of 1000 to 500000 Mw.

The structural formula of the hyaluronic acid and carboxymethyl cellulose is as follows.

[Formula 1]

Structural formula of hyaluronic acid and carboxymethyl cellulose

The state in which the hyaluronic acid and the carboxymethyl cellulose are mixed is called the hyaluronic acid and the carboxymethyl cellulose complex.

The content of the hyaluronic acid and the carboxymethyl cellulose is not limited, and may be varied in consideration of decomposition properties in vivo. As the ratio of carboxymethyl cellulose increases, the degradation rate in vivo decreases.

In vivo resolution is superior to the hyaluronic acid, and in vivo resistance is superior to the carboxymethyl cellulose, the hyaluronic acid of the hyaluronic acid and carboxymethyl cellulose composite film to prevent adhesion and decompose in vivo after a certain time is 1 ~ 80 parts by weight, the carboxymethyl cellulose is characterized in that 1 to 400 parts by weight, preferably 5 to 60 parts by weight, and 5 to 250 parts by weight, respectively.

The crosslinking activator is 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), 1-ethyl-3- (3- (Trimethylammonio) propyl) carbodiimide (1-ethyl-3- (3-trimethylammonio) propyl) carbodiimide, ETC), or 1-cyclohexyl-3- (2-morpholinoethyl) carbodiimide (1 -cyclohexyl-3- (2-morpholinoethyl) carbodiimide (CMC) is characterized in that any one selected from.

Here, the crosslinking activators are substances that are well soluble in water as carbodiimide compounds for activation of the carboxyl groups of the hyaluronic acid and the carboxymethyl cellulose.

The crosslinking adjuvant is N-hydroxysuccinimide (NHS), 1-hydroxybenzotriazole (1-hydroxybenzotriazol, HOBt), 3,4-dihydro-3-hydroxy-4-oxo -1,2,3-benzotriazine (3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazin, HOOBt), 1-hydroxy-7-azabenzotriazole (1- hydroxy-7-azabenzotriazol, HOAt), or N-hydroxysulfosuccinimide (N-hydroxy sulfo succinimide, Sulfo-NHS).

The derivative form ratio and degree of derivatization may be varied according to the ratio of the crosslinking activator and the crosslinking activity aid.

The following scheme 1 is a derivatization process of hyaluronic acid by a crosslinking activator and a crosslinking activator.

 Derivatization Process of Hyaluronic Acid by Crosslinking Activator and Crosslinking Activator

The carboxyl group of carboxymethyl cellulose is also reacted in the same manner as in Scheme 1 to derivatize the carboxymethyl cellulose.

The hyaluronic acid and carboxymethyl cellulose complex derivative film is formed through the reaction process as in Scheme 1 in a state in which the hyaluronic acid and carboxymethyl cellulose of the structural formula 1 are mixed.

In Scheme 1, when only a crosslinking activator or a low crosslinking activator is used, derivatization in the form of N-acylurea pendant is increased as in (A), while using a crosslinking activator. If a large amount of crosslinking activity adjuvant is used here, derivatization in the form of auto-crosslinking is increased as in (B). Therefore, derivatives having various physical properties, such as swelling degree, adhesion of organs, flexibility, biodegradation rate, are prepared according to the ratio of the crosslinking activator and the crosslinking activator.

The derivative film of the N-acylurea pendant form has a high degree of biodegradation with a low degree of swelling, and the auto-crosslinked derivative film has good biodegradability, a high degree of swelling, and good adhesion to organs. Good sex.

In addition, the present invention, in the hyaluronic acid and carboxymethyl cellulose complex derivative gel, the hyaluronic acid and carboxymethyl cellulose complex derivative gel derivatized by reacting the hyaluronic acid and the carboxymethyl cellulose complex with a crosslinking agent comprising two or more epoxy functional groups to provide.

The structure of the hyaluronic acid and the carboxymethyl cellulose is the same as the formula 1 and using the same hyaluronic acid and carboxymethyl cellulose as in the derivative film.

At this time, the content of the hyaluronic acid and the carboxymethyl cellulose is not limited, and may be varied in consideration of decomposition characteristics in vivo. As the ratio of carboxymethyl cellulose increases, the degradation rate in vivo decreases.

In vivo resolution is superior to the hyaluronic acid, and in vivo resistance is superior to the carboxymethyl cellulose, the hyaluronic acid of the hyaluronic acid and carboxymethyl cellulose complex gel to prevent adhesion and decompose in vivo after a certain time And the hyaluronic acid of the carboxymethyl cellulose composite gel is 1 to 200 parts by weight, and the carboxymethyl cellulose is 1 to 100 parts by weight, preferably 5 to 100 parts by weight and 5 to 50 parts by weight, respectively.

The crosslinking agent is butanediol diglycidyl ether (BDDE), ethylene glycol diglycidyl ether (EGDGE), hexanediol diglycidyl ether (1,4-butandiol diglycidyl ether). 6-hexanediol diglycidyl ether, propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, polytetramethylene glycol diglycidyl ether ether), neopentyl glycol diglycidyl ether, polyglycol polyglycidyl ether, diglycerol polyglycidyl ether, glycerol polyglycidyl ether glycerol polyglycidyl ether, tri-methylpropane polyglycidyl ether, non Epoxypropoxyethylene (1,2- (bis (2,3-epoxypropoxy) ethylene), pentaerythritol polyglycidyl ether or sorbitol polyglycidyl ether It is characterized by.

The crosslinking agent including two or more epoxy functional groups is formed by crosslinking by forming an ether bond by reacting with the alcohol group of the hyaluronic acid and the carboxymethylcellulose complex.

That is, the hyaluronic acid and the carboxymethyl cellulose complex derivative gel is characterized by being capable of high temperature and high pressure sterilization while showing excellent viscoelasticity and heat and enzyme stability through an appropriate epoxide reaction in the hydroxyl group.

The following scheme 2 shows the derivatization process of hyaluronic acid by epoxide.

Derivatization Process of Hyaluronic Acid by Epoxide

The alcohol group of the carboxymethyl cellulose is reacted in the same manner as in Scheme 2 to derivatize the carboxymethyl cellulose.

That is, the hyaluronic acid and the carboxymethyl cellulose complex derivative gel are formed through the reaction process as in Scheme 2 in a state in which the hyaluronic acid and the carboxymethyl cellulose of the structural formula 1 are mixed.

The hyaluronic acid and the carboxymethyl cellulose composite derivative gel have a high degree of swelling, good flexibility, and excellent biodegradability.

The hyaluronic acid and carboxymethyl cellulose complex derivatives are formed in the form of films and gels, but the form of the derivatives is not particularly limited and may be varied in form depending on the medically applicable range.

In addition, the hyaluronic acid and carboxymethyl cellulose composite derivative film production method of the present invention, the hyaluronic acid and carboxymethyl cellulose composite derivative film production method comprises the steps of dissolving hyaluronic acid and carboxymethyl cellulose in distilled water; Stirring the dissolved hyaluronic acid and carboxymethylcellulose to form a film by drying; Adding a crosslinking activator and a crosslinking activity aid to the aqueous organic solvent and then mixing the crosslinking reaction on the surface of the film; Purifying the surface crosslinked film using a washing solvent; Drying the purified film; And sterilizing the dried film by gamma irradiation.

First, a film is prepared with the hyaluronic acid and the carboxymethyl cellulose complex, and then the surface of the film is derivatized. At this time, the film is not broken because fine pores are not formed, the physical support is excellent and flexible.

In addition, the organic solvent may be a C ₂ to C ₆ alcohol or an aqueous alcohol solution, such as DMSO, DMF, acetonitrile, methanol, THF, acetone, acetone aqueous solution, ethanol.

The derivative form ratio and degree of derivatization may be varied according to the ratio of the crosslinking activator and the crosslinking activator.

In addition, a derivative film having a superior N-acylurea pendant form using 0.01 to 500 parts by weight of the crosslinking activator and 0.01 to 200 parts by weight of the crosslinking activator with respect to 100 parts by weight of the repeating unit of the hyaluronic acid and the carboxymethylcellulose complex. Alternatively, 0.01 to 500 parts by weight of the crosslinking activator and 40 to 1000 parts by weight of the crosslinking activator are used to form a derivative film having an auto-crosslinked form.

The derivative film of the N-acylurea pendant form has a high degree of biodegradation with a low degree of swelling, and the auto-crosslinked derivative film has good biodegradability, a high degree of swelling, and good adhesion to organs. Good sex.

And, in the method of producing a hyaluronic acid and carboxymethyl cellulose complex derivative gel of the present invention, the hyaluronic acid and carboxymethyl cellulose complex derivative gel production method comprises the steps of dissolving hyaluronic acid and carboxymethyl cellulose in an aqueous alkali solution; Reacting the hyaluronic acid with a carboxymethyl cellulose aqueous alkali solution by adding a crosslinking agent; Washing the crosslinked hyaluronic acid and carboxymethyl cellulose with distilled water; Swelling the washed reactant in physiological saline to form a gel; And sterilizing the gel at high temperature and high pressure.

Here, the amount of the crosslinking agent is 0.01 to 200 parts by weight, preferably 0.1 to 50 parts by weight, based on 100 parts by weight of the hyaluronic acid and the carboxymethyl cellulose composite repeating unit.

The hyaluronic acid and the carboxymethyl cellulose complex derivative gel are excellent in biodegradability, flexible, and have a high degree of swelling.

In addition, the hyaluronic acid and carboxymethylcellulose complex derivatives prepared by the present invention may be variously separated and / or purified by conventional methods known in the art, and such representative examples include distillation (distillation under atmospheric pressure and Distillation under reduced pressure), recrystallization, column chromatography, ion exchange chromatography, gel chromatography, affinity chromatography, thin layer chromatography, phase separation, solvent extraction, dialysis, washing and the like. In this case, the separation and / purification may be carried out after each process or after a series of reactions in the preparation of hyaluronic acid derivative microbeads.

In addition, starting materials and reagents necessary for the synthesis of the hyaluronic acid and carboxymethylcellulose complex derivatives of the present invention can be easily prepared by the methods of the literature or by the aforementioned methods, or can be commercially purchased.

As described above, the hyaluronic acid and carboxymethylcellulose composite derivative films and gels having the organ adhesion prevention properties according to the present invention, and a method for producing the same are excellent in biodegradability, have high flexibility, and are initially used by surface derivatization methods. Maintains stable form, prevents adhesion and completely decomposes and absorbs. In addition, while preventing the adhesion after surgery, within four weeks of decomposition, metabolism, release is less side effects, good adhesion to the tissue, flexible, easy to use, less pore, the physical strength is large.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited by the following examples.

Example 1 Preparation of Hyaluronic Acid and Carboxymethyl Cellulose Composite Films According to Concentrations of Various Crosslinking Activators / Crosslinking Aids

167 mg hyaluronic acid and 83 mg carboxymethylcellulose complex were dissolved in 12.5 ml distilled water. The mixture was stirred for a certain time and then dried to prepare a film. The prepared film was reacted in a 25 ml ethanol solution in which 62.5 mg, 125 mg, 250 mg of EDC and 75 mg, 150 mg, and 300 mg of NHS were dissolved. After the reaction, an unreacted substance was removed using ethanol to prepare a film in which derivatization proceeded only on the surface. The dried film was sterilized by gamma irradiation at 25 KGy. The prepared film was confirmed structural analysis through FT-IR.

IR Analysis of Hyaluronic Acid and Carboxymethylcellulose Complex Derivative Films Amount of crosslinking activator / crosslinking auxiliary 0mg / 0mg 62.5mg / 75mg 125mg / 150mg 250mg / 300mg A ₁₇₅₀ / A ₁₆₀₀ 0.024 0.03 0.096 0.156 A ₁₇₀₀ / A ₁₆₀₀ 0.093 0.183 0.352 0.667

In Table 1, A ₁₇₅₀ is the absorbance of the C = O band of the ester bond, A ₁₇₀₀ is the absorption of C = O band in the form of N-acylurea pendant of EDC crosslinking activator, A ₁₆₀₀ is in the salt form The absorptivity of C = O band of the carboxyl group which exists is shown. In Table 1, A ₁₇₅₀ / A ₁₆₀₀ shows the degree of auto-crosslinking, and A ₁₇₀₀ / A ₁₆₀₀ shows the degree of N-acylurea pendant form of EDC, which is a crosslinking activator. As the amount of active aid increases, the relative value of absorbance also increases.

Experimental Example 1 Swelling degree of hyaluronic acid and carboxymethyl cellulose composite derivative film according to the concentration of various crosslinking activator / crosslinking aid

10 mg of each film prepared in Example 1 was poured into 10 ml of physiological saline to swell. After standing at room temperature for 24 hours, the degree of swelling was determined by dividing the swollen weight by the dry weight.

Swelling degree of hyaluronic acid and carboxymethyl cellulose composite derivative film Amount of crosslinking activator / crosslinking auxiliary 62.5mg / 75mg 125mg / 150mg 250mg / 300mg W _wet / W _dry 13.58 6.99 3.99

In Table 2, the swelling degree of the hyaluronic acid and the carboxymethyl cellulose composite derivative film is decreased while the amount of the crosslinking activator and the crosslinking activity assistant is increased.

Example 2 Preparation of Films According to Various Hyaluronic Acid and Carboxymethyl Cellulose Composite Compositions

167 mg / 83 mg (HSF_21), 125 mg / 125 mg (HSF_11) and 50 mg / 200 mg (HSF_14) of hyaluronic acid and the carboxymethylcellulose complex were dissolved in 12.5 ml distilled water, respectively. The mixture was stirred for a certain time and then dried to prepare a film. The prepared film was reacted in 25 ml ethanol solution in which 250 mg of EDC and 300 mg of NHS were dissolved. After the reaction, the ethanol was used to remove the unreacted material, and a film having derivatization only on the surface thereof was prepared. The dried film was sterilized by gamma irradiation at 25 KGy. The prepared film was confirmed structural analysis through FT-IR.

IR analysis of various hyaluronic acid and carboxymethylcellulose complex derivative films Hyaluronic Acid / Carboxymethyl Cellulose 167mg / 83mg 125mg / 125mg 50mg / 200mg A ₁₇₅₀ / A ₁₆₀₀ 0.156 0.283 0.270 A ₁₇₀₀ / A ₁₆₀₀ 0.667 0.452 0.408

Experimental Example 2 In vitro Enzyme Resistance Test of Films Prepared According to Various Hyaluronic Acid and Carboxymethyl Cellulose Complex Compositions

Each film prepared in Example 2 and 10 mg of the commercialized S film of G Company was put in 10 ml of physiological saline to swell. 1000 units of hyaluronic acid degrading enzyme was added to each swollen film at regular time intervals, and then stored at 37 ° C. The degree of degradation in vitro was confirmed by checking the weight of the swollen film at regular time intervals.

1 is a diagram showing in vitro enzyme resistance of a film prepared according to various hyaluronic acid and carboxymethyl cellulose complex composition. In Figure 1, it shows that the resistance to the enzyme is increased as the content of the carboxymethyl cellulose in the composition of the hyaluronic acid and carboxymethyl cellulose complex. In addition, derivatization only on the surface maintains the film having a stable structure earlier than the overall derivatization of the S film of G Company, and exhibits decomposition characteristics that are rapidly decomposed at a certain point in time. This shows that derivatization focused only on the surface maintains an initial stable state in vivo, and relatively low derivatization inside the film shows a structure that can be easily decomposed and absorbed. It shows flexibility and maintains a stable form at the beginning, and shows excellent adhesion prevention effect with decomposition property that is completely decomposed and absorbed after preventing adhesion phenomenon.

Example 3 Preparation of Hyaluronic Acid and Carboxymethyl Cellulose Complex Derivative Gels by Epoxide

667 mg of hyaluronic acid and 333 mg of carboxymethylcellulose were dissolved in 0.25 N NaOH solution to prepare 10 ml. To the solution was added 50 mg of butanediol diglycidyl ether (BDDE). After reacting at room temperature for 24 hours, the reaction product was washed with distilled water to remove unreacted material. The washed reaction was swollen in physiological saline and sterilized at 121 ° C. to prepare hyaluronic acid and carboxymethylcellulose complex derivative gel.

Experimental Example 3 Comparison of Physical Properties of Hyaluronic Acid and Carboxymethyl Cellulose Complex Derivative Gel by Epoxide

In order to confirm the rheological properties of the hyaluronic acid and carboxymethylcellulose complex derivative gels prepared in Example 3 and the commercialized H gels and hyaluronic acid gels, the rheometer was used at a frequency of 0.02 to 1 Hz. After sterilization, complex viscosity, elasticity and viscosity coefficient were measured.

Figure 2 is a view comparing the physical properties of the hyaluronic acid and carboxymethyl cellulose complex derivative gel, as shown in Figure 2, the hyaluronic acid and carboxymethyl cellulose complex derivative gel is better composite viscosity than the H gel commercialized as an anti-adhesion agent The value is showing. In addition, it is shown that the formation of a network with high structural stability through the elastic modulus and viscosity coefficient values.

Experimental Example 4 Confirmation of anti-adhesion effect of hyaluronic acid and carboxymethyl cellulose complex derivative film through animal model

A model of abrasion was made on the cecum and abdominal wall of rats and commercialized with a control group without using an anti-adhesion agent and a hyaluronic acid and a carboxymethylcellulose complex derivative film prepared in Example 2 (using a crosslinking activator 125mg / crosslinking activity adjuvant, HSF_11). The change of adhesion was observed in the experimental group of the S film of G Company. For these experiments, Sprague Dawley rats weighing 240-270 g were used. Each of the five animals in the control and experimental groups were damaged. Abdominal incisions were made after anesthesia with dilutions of 5 mg / 100 g of ketamine and 10 mg of ampicillin injected by the intramuscular route of each animal. The cecum was injured to the extent of bleeding to a size of 12 cm. After the same amount of damage was applied to the opposite peritoneal membrane, the experimental group was cut into a film of sufficient size to be sandwiched between the cecum and the abdominal wall. Two sites were corrected with sutures 1 cm away from the site of injury. Two weeks after the operation, the degree of adhesion, adhesion strength, and adhesion area were evaluated. In the evaluation of adhesions, the degree of adhesion was evaluated by the following score.

0. No adhesion

1. One thin film type adhesion

2. Two or more thin film type adhesions

3. Spotted concentrated thick coalescence

4. Plated concentrated thick coalescence

5. very thick adhesions with blood vessels or one or more platy adhesions

In addition, the adhesion intensity in the evaluation of adhesion was evaluated by the following score.

0. No adhesion

1.Adhesion that is film-type and drops even with very weak force

2.Adhesion requiring medium strength

3. Cohesion that can be released only when a considerable pressure is applied

4.If the adhesion is very strong and difficult to peel off or very high pressure is required

As shown in Figures 3 to 6, the hyaluronic acid and carboxymethyl cellulose complex derivative film of the present invention serves to prevent the formation of adhesions between adjacent tissues by acting as a physical barrier during the healing of tissue wounds .

As shown in Figure 3, the experimental group using the hyaluronic acid and carboxymethyl cellulose complex derivative film (HSF_11) of the present invention compared to the control group was less adhesion to the tissue appeared to be separated from the surrounding tissue.

As shown in Figure 4, the degree of adhesion of the experimental group using the hyaluronic acid and the carboxymethyl cellulose composite derivative film of the present invention (HSF_11) was about 1.2 compared to the experimental group of the control group of 4.8 and the S film of G company of 1.4. The degree of adhesion of 1.2 is expressed as a single thin film type adhesion.

As shown in Figure 5, the adhesion strength of the experimental group using the hyaluronic acid and carboxymethyl cellulose composite derivative film of the present invention (HSF_11) was about 0.8 compared to the experimental group of the control group of 3.8 and S film of G company of 1.2. The adhesion strength of 0.8 is a film form and is expressed by adhesions that fall even with very weak forces.

As shown in Figure 6, the adhesion area of the experimental group using the hyaluronic acid and carboxymethyl cellulose composite derivative film (HSF_11) of the present invention was about 0.1 cm 2 compared to the experimental group of the control film of 2.3 cm 2 and S film of G company of 0.2 cm 2 . Since the adhesion area of the composite derivative film of the present invention is small, the derivative film of the present invention can be easily separated, biodegradable, metabolized, and released due to less adhesion to tissues after being used as a barrier.

In particular, the hyaluronic acid and the carboxymethyl cellulose composite derivative film of the present invention exhibited excellent characteristics of weaker adhesion strength and less adhesion area than the commercialized S film of G Company.

The above description is merely illustrative of the technical idea of the present invention, and those skilled in the art to which the present invention pertains may make various modifications and changes without departing from the essential characteristics of the present invention. Therefore, the embodiments disclosed in the present invention are not intended to limit the technical idea of the present invention but to describe the present invention, and the scope of the technical idea of the present invention is not limited by these embodiments. The protection scope of the present invention should be interpreted by the following claims, and all technical ideas within the equivalent scope should be interpreted as being included in the scope of the present invention.

1 is a diagram showing in vitro enzyme resistance of a composite derivative film prepared according to various hyaluronic acid and carboxymethyl cellulose complex composition.

Figure 2a is a diagram comparing the composite viscosity of the hyaluronic acid and carboxymethyl cellulose complex derivative gel, Figure 2b is a view comparing the elastic modulus and viscosity of the hyaluronic acid and carboxymethyl cellulose complex derivative gel.

3 is a photographic view applied to an animal in a rat animal model.

4 is a view showing the degree of adhesion in the rat animal model.

5 shows adhesion strength in a rat animal model.

Fig. 6 shows the adhesion area in the rat animal model.

Claims (9)

A hyaluronic acid and a carboxymethyl cellulose composite derivative film in which an N-acylurea pendant form and an auto-crosslinking form are mixed by derivatization of a hyaluronic acid and a carboxymethyl cellulose composite film by reacting a crosslinking activator and a crosslinking activator. According to claim 1, N-acylurea pendant form using 0.01 to 500 parts by weight of the crosslinking activator and 0.01 to 200 parts by weight of the crosslinking activator with respect to 100 parts by weight of the repeating unit of the hyaluronic acid and carboxymethyl cellulose complex Hyaluronic acid and carboxymethylcellulose, characterized in that to form a predominant derivative film, or to form a derivative film having an auto-crosslinking form using 0.01 to 500 parts by weight of the crosslinking activator and 40 to 1000 parts by weight of the crosslinking activator. Composite derivative film. The method of claim 1, wherein the crosslinking activator is 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), 1-ethyl -3- (3- (trimethylammonio) propyl) carbodiimide (1-ethyl-3- (3-trimethylammonio) propyl) carbodiimide, ETC), or 1-cyclohexyl-3- (2-morpholinoethyl ) Carbodiimide (1-cyclohexyl-3- (2-morpholinoethyl) carbodiimide, CMC) hyaluronic acid and carboxymethyl cellulose complex derivative film, characterized in that any one selected from. The method of claim 1, wherein the crosslinking adjuvant is N-hydroxysuccinimide (NHS), 1-hydroxybenzotriazol (1-hydroxybenzotriazol, HOBt), 3,4-dihydro-3- Hydroxy-4-oxo-1,2,3-benzotriazine (3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazin, HOOBt), 1-hydroxy-7-aza Hyaluronic acid and carboxymethylcellulose, characterized in that any one selected from benzotriazole (1-hydroxy-7-azabenzotriazol, HOAt), or N-hydroxysulfosuccinimide (Sulfo-NHS) Composite derivative film. A hyaluronic acid and a carboxymethyl cellulose complex derivative gel derivatized by reacting a hyaluronic acid and a carboxymethyl cellulose complex with a crosslinking agent comprising two or more epoxy functional groups. According to claim 5, The hyaluronic acid and the hyaluronic acid of the carboxymethyl cellulose composite gel is 1 to 200 parts by weight, the carboxymethyl cellulose is 1 to 100 parts by weight, preferably 5 to 100 parts by weight, respectively, 5 ~ 50 parts by weight of hyaluronic acid and carboxymethylcellulose complex derivative gel. The method of claim 5, wherein the crosslinking agent is butanediol diglycidyl ether (BDDE), ethylene glycol diglycidyl ether (EGDGE), hexanediol diggle Lycidyl ether (1,6-hexanediol diglycidyl ether), propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, polytetramethylene glycol diglyci Polytetramethylene glycol diglycidyl ether, neopentyl glycol diglycidyl ether, polyglycol polyglycidyl ether, diglycerol polyglycidyl ether, Glycerol Polyglycidyl Ether, Trimethyl Propane Polyglycidyl Ether glycidyl ether, bisepoxypropoxyethylene (1,2- (bis (2,3-epoxypropoxy) ethylene), pentaerythritol polyglycidyl ether or sorbitol polyglycidyl ether Hyaluronic acid and carboxymethyl cellulose complex derivative gel, characterized in that any one selected from. Dissolving hyaluronic acid and carboxymethylcellulose in distilled water; Stirring the dissolved hyaluronic acid and carboxymethylcellulose to form a film by drying; Adding a crosslinking activator and a crosslinking activity aid to the aqueous organic solvent and then mixing the crosslinking reaction on the surface of the film; Purifying the surface crosslinked film using a washing solvent; Drying the purified film; And Sterilizing the dried film by gamma irradiation; hyaluronic acid and carboxymethyl cellulose complex derivative film production method comprising a. Dissolving hyaluronic acid and carboxymethylcellulose in an aqueous alkali solution; Reacting the hyaluronic acid with a carboxymethyl cellulose aqueous alkali solution by adding a crosslinking agent; Washing the crosslinked hyaluronic acid and carboxymethyl cellulose with distilled water; Swelling the washed reactant in physiological saline to form a gel; And Sterilizing the gel at a high temperature and high pressure.

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