WO2012144678A1 - Drug delivery composition containing biocompatible cross-linked hyaluronic acid product - Google Patents

Abstract

The present invention provides a drug delivery composition comprising: a protein drug or an anti-inflammatory drug in a carrier or a support essentially comprising a polysaccharide such as a polysaccharide and the like used in a pharmaceutical dosage form such as in an injectable form and the like or in a preparation for tissue engineering; and a cross-linked product of hyaluronic acid, a polysaccharide, and a biocompatible polymer for controlling the release amount. According to the present invention, the drug delivery composition can show an extended release property, improves tissue engineering healing, is safe, and can reduce manufacturing costs.

Description

Drug Delivery Compositions Including Biocompatible Hyaluronic Acid Crosslinks

The present invention relates to a composition comprising a biocompatible hyaluronic acid crosslinked product.

Hyaluronic acid began in 1934 by extracting a previously unknown new ingredient from the vitreous of the bovine eye. The name hyaluronic acid is used to mean hyaloid and uronic acid, and it is an important mucopolysaccharide such as chondroitin sulfate. This component is present in animal vitreous, joint fluid, cartilage, skin, etc. It is combined with a large amount of water and becomes a gel state. It is involved in lubrication of joints and flexibility of the skin. It is also known to play an important role in preventing skin penetration. In skin tissues, hyaluronic acid is a gel-like substance between collagen, elastin, and fibrous tissue. Hyaluronic acid is a natural component of living organisms, especially in many parts of the human body. In particular, it exists in the skin to make the skin elastic and make volume. If hyaluronic acid is insufficient, the skin becomes dry, elastic and wrinkled. Recently, it is widely used as an injection medicine to prevent wrinkles. Collagen and hyaluronic acid are two typical filters to replenish the volume of boned skin such as wrinkles. In addition, these two types are the most commonly used as a filter and are made by injection or capsule or cosmetics. Hyaluronic acid is also good for the skin, but plays a more important role in the treatment of osteoarthritis. Hyaluronic acid is part of the cartilage synovial fluid secreted from the joints to soften the cartilage, which buffers the bones between the bones and the joints. In cartilage tissues, hyaluronic acid helps to bond between molecules and reinforces cartilage to withstand joint movement and weight.

Hyaluronic acid is a linear polysaccharide composed of glucuronic acid and acetylglucosamine, which is present in the extracellular matrix (ECM), synovial fluid of joints, and scaffolds that make up cartilage. One of the glycosamino glycans. Hyaluronic acid also plays an important role as a signaling molecule in cell motility, cell differentiation, wound healing and cancer metastasis. The unique viscoelastic properties of hyaluronic acid crosslinked with hyaluronic acid add to its importance as joint lubricants. In addition, since hyaluronic acid has no problem in terms of immunity, it is a biomaterial having excellent biocompatibility that can be used for tissue engineering and drug delivery system. Hyaluronic acid and hyaluronic acid oligosaccharides have a three-dimensional structure in solution, which causes a wide range of internal hydrogen bonds, limited fluidity of the polymer chains, and unique helical and coiled coil reactions. do.

Hydrogels interact by chemical bonds or electrostatic forces to form hydrophilic cross-linked polymers and absorb water in amounts ranging from several hundred times their dry weight. Hydrogels are known to be widely used in the pharmaceutical and medical fields due to their excellent biocompatibility and hydrophilic properties. Hydrogels, on the other hand, are excellent candidates for the manufacture of tissue engineering substrates aimed at healing damaged human tissue. Tissue engineering is a substantially new field of science dealing with the development of techniques useful for achieving regeneration of damaged tissues or reproduction of humans. Cells adhere and proliferate through three-dimensional structures, particularly porous microstructures, to allow for the growth and differentiation of restoring tissue cells, such as fibroblasts, chondrocytes, osteoblasts, etc., and deposition of extracellular matrix. It can form new extracellular matrix. These three-dimensional structures are commonly referred to as scaffolds and hydrogels are suitable for constructing similar substrates for such tissue engineering. Particularly, the hydrogel's properties allow for excellent flow of nutrients into cells and product flow outside the cells to promote cell adhesion, proliferation and growth in hydrogels. The main characteristics are the possibility of modifying the surface by bonding.

Hyaluronic acid generally has a molecular weight of about 1,000 ~ 10,000,000 Da and has the unique physicochemical properties and specific biological functions as mentioned above.

Hyaluronic acid plays a major role in hemostasis of the tissues and lubrication of the joints, and it binds to specific proteins on the surface of the cells and plays a very important role in cell fluidity, growth factors, and inflammatory reactions. do.

Tissue engineering applications include the possibility of using biodegradable sponges or biodegradable films for joint cartilage regeneration or to protect or heal wounds caused by burns, diseases. In this case, hydrogel may be applied to the wound to support rapid regenerative activity, and these fibroblasts rapidly synthesize new ECM and heal the wound after attaching to the skeleton. Similar skin disease agents perform temporary protective functions that can reduce the risk of infection from wounds. Skeletons that are selectively filled with drugs can perform drug delivery and, for example, delay the release of antibiotics or growth factors depending on the prevalence of the wound. Hyaluronic acid is a basic component of the extracellular matrix widely distributed in animal tissues. Hyaluronic acid functions to regulate cell proliferation and differentiation. It is involved in important biological processes, the aforementioned cell mobility and tissue healing, which regulate the inflammatory response. Hyaluronic acid has been shown to be involved in tumor growth by interacting with specific receptors located on the cell surface. This draws much attention to the possibility of the modified hyaluronic acid being applied as a soluble carrier in the production of macromolecular prodrugs having antitumor activity.

From the above, the hyaluronic acid-based biomaterial is very suitable as a tissue support for biocompatibility and cell growth induction. Nevertheless, hyaluronic acid alone is not very elastic and has a disadvantage in that it easily breaks down. Limited. In addition, the surfaces of these skeletons are so hydrophilic that they have poor adhesion and cell differentiation. For this reason, a method of modifying hyaluronic acid by mixing or crosslinking hyaluronic acid with biocompatible natural polymers such as collagen and gelatin, or by modifying hyaluronic acid with a hydrophobic group has been proposed. Similarly, by introducing functional groups on the surface of hyaluronic acid, chemical modification in polysaccharide molecules aims to obtain a drug delivery system (DDS) that slows the release rate of drugs introduced into hyaluronic acid. The drug can be transported to the site of action in the state of being directly chemically bound to hyaluronic acid and can increase its bioavailability, or it can be physically enclosed in chemically modified hyaluronic acid and according to the characteristics of the modified hyaluronic acid. It is a system that can control the release mechanism of drug. Unmodified hyaluronic acid, by itself, plays an important role in drug delivery and surgical operations. Hyaluronic acid plays a role as an adjuvant in ophthalmic drug release (Mucoadhesive ophthalmic vehicles: evaluation of polymeric low-viscosity formulations, J. Ocular Pharm. 10 (1994) 83-92). It has also been found to enhance drug and protein absorption through mucosal tissues (Efficacy of hyaluronic acid / nonsteroidal anti-in-flammatory drug systems in preventing postsurgical tendon adhesions, J. Biomed. Mater. Res. (Appl. Biomater.) 38 (1997) 25-33). Hyaluronic acid is a biocompatible, biodegradable polymeric material that is likely to be used in drug delivery systems and tissue engineering applications. While maintaining the biocompatibility, biodegradability, and non-immunity properties of hyaluronic acid itself, the physical and chemical properties can be modified according to the applied situation by improving the physicochemical properties through chemical modification and changing the resolution in vivo. have. The composition of hyaluronic acid and the reaction of carboxyl groups, polymer-drug coupling for controlling drug release mechanisms, crosslinking with hydrogels, surface coating, and the like and other compositions were also discussed. Many hyaluronic acid derivatives (eg, self-crosslinked hyaluronic acid) have been evaluated in vitro and in vivo. The esterification of hyaluronic acid with various drugs is known to provide new physicochemical properties that can be used for drug release control and tissue engineering.

It is not easy to obtain hyaluronic acid derivatives that maintain sufficient strength when applied in vivo. The best method is to increase the crosslinking of hyaluronic acid at temperature conditions that do not cause drug or cell problems in the polymer hydrogel manufacturing process.

A small amount of crosslinking agent must be used to obtain biomaterials for physiological use and sufficient purification is required when using an excess of crosslinking agent. It is also important that the crosslinking agent used does not itself cause toxicity to the living body. Background Art Many biocompatible polymer materials have been of interest as carriers for efficiently transporting drugs into the human body. Various techniques have been developed for the purpose of increasing the bioabsorption rate of the existing drug, or targeted drug delivery to release the drug specifically to the required area.

Methods for crosslinked hyaluronic acid derivatives have been proposed in many literatures. That is, a method of preparing a compound having two functional groups, such as divinyl sulfone, bisepoxide, bishalide, formaldehyde, and the like, has been reported. US Pat. No. 4,458,865 uses divinyl sulfone for crosslinking hyaluronic acid. An example is disclosed and US Patent No. 4713448 reports crosslinking reactions using formaldehyde. Because of the toxicity and unreacted substances of unreacted chemicals, they are not suitable for use as high purity biocompatible materials. U.S. Patent No. 5356883 discloses a synthesis example of a hyaluronic acid derivative hydrogel in which a carboxyl group is modified with O-acylurea or N-acylurea using various carbodiimides. However, the hyaluronic acid cross-linked product prepared by the method of these patents has a problem of low stability to degrading enzymes. U.S. Patent Application Publication No. 2006 / 0105022A1 discloses the preparation of hyaluronic acid crosslinked products that exhibit excellent viscoelasticity while using excessive hyaluronic acid to improve physical properties and lowering the ratio of crosslinking agents. In this method, a large amount of crosslinking agent is added to increase the crosslinking rate, thereby focusing on increasing viscoelasticity and increasing long-term tissue correction. In this case, there is a disadvantage in that it is not easy to manufacture at high concentration. In addition, Korean Patent Publication No. 10-2009-0013696 discloses a hyaluronic acid crosslinked hydrogel by reacting hyaluronic acid or a salt or derivative thereof with a polyhydrazide compound of an aliphatic polycarboxylic acid in the presence of a carboxyl activator in a water-alcohol mixed solvent. It has been presented a method of preparation. Luo et al. (Cross-linked hyaluronic acid hydrogel films: new biomaterials for drug delivery, Journal of Controlled Release 69 (2000) 169-184) Hyaluronic acid was first made into adipic dihydrazide derivatives and then hyaluronic acid using polyethylene glycol-propionaldehyde Lonic acid was prepared with a crosslinked hydrogel and the drug release mechanism of antibacterial drugs such as hydrocortisone, prednisolone, cortisone, dexamethasone, or anti-inflammatory agent was confirmed from the crosslinked hyaluronic acid hydrogel. Lee et al. (An injectable hyaluronic acid-tyramine hydrogel system for protein delivery, Journal of Controlled Release 134 (2009) 186.193). The oxidation of tyramine with hydrogen peroxide (H ₂ O ₂ ) and horseradish peroxide (HRP) An injectable hyaluronic acid hydrogel system conjugated with tyramine was developed and designed to control the release rate of protein drugs such as lysozyme or amylase in the hyaluronic acid gel. However, these methods require several chemical reactions to the final material to be targeted, so there is still a problem that is difficult to apply directly to commercialization.

In Korean Patent 10-0818659, a free sulfate group is introduced by using a carboxyl group of 10000-2x107 molecular weight hyaluronic acid, dissolved in an aqueous solution with a drug, and a water in oil reverse emulsion is formed by disulfide bonds. It is known to manufacture a self-crosslinked hyaluronic acid nanogel of 100 ~ 900nm diameter.

On the other hand, protein drugs are useful for the treatment of various diseases, but the short half-life and low absorption rate in vivo has a limit in producing a therapeutic effect. Due to the low body absorption rate, most of the injections are administered, and the half-life of the body is short as 2 ~ 4 hours even during the injection. Therefore, repeated drug administration is required. Regarding macromolecular drugs such as protein preparations, biodegradable synthetic polymers such as polylactic acid and polyglycolic acid are used to prepare micro / nanoparticles using water in oil emulsion, or polyethylene glycol PEGylation with (polyethylene glycol) or complexes between polymer materials using anions and cations are used. However, in the case of microparticles prepared using polylactic acid or polylactide-glycolic copolymers, there is a disadvantage in that degeneration of protein drugs is caused due to the hydrophobicity of the polymer material. The resulting acid lowers the pH and promotes denaturation and aggregation of the drug. Norpeare's method is less efficient because of low drug loading and low drug delivery. PEGylation also has low delivery efficiency and negative effects on cytotoxicity when using ionic polymers and does not support safety in immune responses. From this point of view, natural polymers such as hyaluronic acid can be used as drug carriers, but in itself, their viscosity and density are drastically reduced due to decomposition in vivo, and thus drug retention activity is lost.

Glen et al. (Controlled chemical modification of hyaluronic acid: synthesis, applications, and biodegradation of hydrazide derivatives, Journal of controlled release 53 (1998) 93-103). The release behavior has been suggested by chemical bonding. In general, the higher the viscosity of hyaluronic acid, the more effective the sustained release of the drug. However, in the case of a composition having a high content of hyaluronic acid, the viscosity is so high that it is difficult to administer to the patient. In Japanese Patent Laid-Open No. 1987-287041, it has been reported that when insulin is injected into a rabbit, an insulin preparation containing hyaluronic acid does not sustain a glucose-lowering effect by insulin. In addition, U. S. Patent No. 5416071 discloses a sustained release formulation of interferon, including hyaluronic acid and plasma protein, but reported that blood concentration rapidly decreased to 1/10 of the initial concentration within 24 hours. In U.S. Patent Publication No. 2003/0064105, hyaluronic acid microparticles were prepared, coated with a lipophilic substance such as lecithin, and then dispersed in oil to control drug release. In addition, oil in water of lecithin-coated microparticles was controlled. emulsion). Therefore, there is a need for the development of formulations and compositions that exhibit excellent sustained release effects over a period of time while maintaining the physiological activity of protein drugs.

The present invention was created to solve the problems of the prior art as described above, it is possible to physically enclose a protein drug or anti-inflammatory drug and to control the drug release, such as inducing drug release for a certain period of time to effect the effect of the drug It is an object of the present invention to provide an excellent drug delivery composition including a hydrogel by cross-linking of hyaluronic acid polymer that maximizes and solves the cytotoxicity problem generated during crosslinking.

In one aspect of the present invention, a drug delivery composition comprises a protein drug or an anti-inflammatory drug in a carrier or a support, which essentially includes a polysaccharide, such as hyaluronic acid, which is used as a pharmaceutical formulation such as an injection or a formulation for tissue engineering. And a crosslinked product with hyaluronic acid, a polysaccharide, and a biocompatible polymer in order to control and control the release amount thereof.

The molecular weight of the hyaluronic acid is characterized in that 30,000 Daltons (Da) to 3,000,000 Daltons (Da).

The drug delivery composition comprises a carboxyl group of hyaluronic acid and other biocompatible polymers and biodegradable polymers using EDC (1-Ethyl-3- (3-dimethylaminopropyl) -carbodiimide) in a 0.5 to 10% by weight aqueous solution of hyaluronic acid. Hyaluronic acid hydrogels crosslinked by an amide reaction with any one selected from the group; And protein drugs or nonsteroidal anti-inflammatory agents; It is characterized by containing.

The biocompatible polymer is characterized in that selected from the group consisting of methyl cellulose, carboxymethyl cellulose, hydroxy propylmethyl cellulose, alginate, chitosan, gelatin, collagen and mixtures thereof or derivatives thereof.

The biodegradable polymer is characterized in that it is selected from homopolymers of lactic acid having a molecular weight ranging from 6,000 to 100,000, copolymer derivatives of lactic acid and glycolic acid, or mixtures thereof.

The protein drug is characterized in that selected from the group consisting of albumin, insulin, erythropoietin, insulin growth factor, platelet-derived growth factor, modified growth factor alpha, modified growth factor beta, bone-forming protein and mixtures thereof.

The anti-inflammatory drug is characterized in that it is selected from the group consisting of pyroxicam, meloxium, ibuprofen, metoprofen, diclofenac, indomethacin, tetracycline, monocycline, doxycycline and mixtures thereof.

The content of hyaluronic acid in the cross-linked hyaluronic acid hydrogel is characterized in that 30 to 80% by weight.

As described above, the present invention has the following effects.

First, the drug delivery composition according to the present invention may exhibit long-term persistence while the protein drug or anti-inflammatory agent maintains biological activity.

Second, the drug delivery composition according to the present invention may be prepared in a formulation such as an injectable preparation.

Third, the drug delivery composition according to the present invention can be prepared in a biocompatible, biodegradable scaffold to control the inflammatory response that may occur when the biomaterial is inserted into the human body as a support for tissue engineering to increase histological healing power.

Fourth, the cytotoxicity of the final composition is safer than the crosslinked hyaluronic acid hydrogel preparation methods that have been proposed previously.

Fifth, the manufacturing process for the final composition is simple and simple compared to other methods, the manufacturing cost can be much reduced.

1 is a graph showing the water content according to the weight fraction of hyaluronic acid in the cross-linked hyaluronic acid / polysaccharide hydrogel component.

Figure 2 is a cytotoxicity test results of hyaluronic acid and cross-linked hyaluronic acid hydrogel prepared by the present invention.

Figure 3 is the result of confirming the amount released after the addition of albumin to the cross-linked hyaluronic acid, hyaluronic acid / gelatin, hyaluronic acid / chitosan, hyaluronic acid / collagen hydrogel over time.

FIG. 4 shows the weight fraction of hyaluronic acid in the crosslinked hydrogel when the hyaluronic acid / gelatin crosslinked hydrogel is prepared according to the present invention. Albumin is released after the albumin is added to each of the prepared crosslinked hydrogels. This is the result of checking the quantity over time.

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

The present invention is described in more detail with reference to the examples, but it is only illustrative of the present invention, and the content of the present invention is not limited by the following examples.

Preparation Example 1

1% (w / v) hyaluronic acid solution was prepared and 50mM EDC (1-Ethyl-3- (3-dimethylaminopropyl) -carbodiimide) was added thereto, and the solution was stirred at 4 ^o C for 24 hours. Thereafter, 1% (w / v) gelatin solution was mixed, stirred well for about 5 minutes, and then crosslinked for 6 hours. pH was adjusted to 7.4.

Preparation Example 2

1% (w / v) hyaluronic acid solution was prepared and 50mM EDC (1-Ethyl-3- (3-dimethylaminopropyl) -carbodiimide) was added thereto, and the solution was stirred at 4 ^o C for 24 hours. Thereafter, a 1% (w / v) chitosan solution was mixed and stirred well for about 5 minutes, followed by crosslinking reaction for 6 hours. pH was adjusted to 7.4.

Preparation Example 3

1% (w / v) hyaluronic acid solution was prepared and 50mM EDC (1-Ethyl-3- (3-dimethylaminopropyl) -carbodiimide) was added thereto, and the solution was stirred at 4 ^o C for 24 hours. Thereafter, the 1% (w / v) collagen solution was mixed and stirred well for about 5 minutes, and then crosslinked for 6 hours. pH was adjusted to 7.4.

Experimental Example 1

The water content of the hyaluronic acid hydrogel crosslinked in the same manner as in Preparation Example 1, Preparation Example 2, Preparation Example 3 was experimentally confirmed by the following method. At this time, the weight fraction of hyaluronic acid in the crosslinked hyaluronic acid hydrogel was changed to 10, 20, 40, 60, 80, 90%, respectively, to prepare a final crosslinked hydrogel. The prepared crosslinked hydrogel was immersed in PBS (phosphate-buffered saline) for 24 hours at 37 ℃. It was taken out at a fixed time and gently wiped dry. The weights of the swollen hydrogels and the dried hydrogels were measured, respectively, and the water content was calculated by the following formula.

[formula]

Water content (%) = [(Swelled hyaluronic acid hydrogel weight-Dried hyaluronic acid hydrogel weight) / Swelled hyaluronic acid hydrogel weight] X 100

As shown in FIG. 1, the water content of the hydrogel was stably increased between 20 to 80% by weight of the hyaluronic acid in the crosslinked hyaluronic acid hydrogel. These experimental results were confirmed in all crosslinked hydrogels such as gelatin, chitosan and collagen. This confirms that the basic swelling characteristics that should be present as drug carriers are relatively well implemented.

Experimental Example 2

Suspension solution containing 1 × 10 4 cells / ml of RAW 264.7 mouse macrophage using medium containing 10% FBS (fetal bovine serum), RPMI (rosewell park memorial institute medium) and 1% penicillin / streptomycin Was prepared. The prepared suspension was injected into each well of 94 wells and incubated for 24 hours. At this time, each well was injected into three wells (n = 3). After 24 hours, the medium was removed from each well, washed with PBS (phosphate buffered saline) solution, and treated with a crosslinked hyaluronic acid sample (100 μl), respectively. Crosslinked hyaluronic acid samples were sterilized by filtering with a 0.22 μm syringe filter. Cytotoxicity experiments were performed 24 hours after treatment with crosslinked hyaluronic acid samples. That is, the cell solution containing the MTT reagent and the crosslinked hyaluronic acid material in each medium was transferred to each well and placed in the incubation for 1 or 2 days. To determine the cytotoxicity due to biomaterials, MTT solution (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazlium bromide) was added and incubated for an additional 4 hours and ELIZA analyzer at 550 nm. Absorbance was measured using. (Figure 2)

Experimental Example 3

For the preparation of drug-encapsulated cross-linked hyaluronic acid hydrogel and drug release experiment, first, an aqueous solution was prepared by using albumin powder at a constant concentration. The crosslinked hyaluronic acid hydrogel was placed therein so that all of the albumin solution was absorbed into the hydrogel. The loading amount of albumin could be adjusted by changing the initial albumin concentration in aqueous solution. Albumin-containing hyaluronic acid hydrogel was dried for 24 hours.

The hyaluronic acid cross-linked hydrogel filled with albumin was placed in PBS at 37 ° C., gently shaken, and a predetermined amount of solution was taken at a predetermined time to measure the amount of albumin released from the hyaluronic acid hydrogel using a UV spectrometer. (Figure 3)

In addition, when the hyaluronic acid / gelatin crosslinked hydrogel is prepared, the weight fraction of hyaluronic acid in the crosslinked hydrogel is changed to 5,20,40,80,90% to prepare a crosslinked hydrogel. Drug release from the gel was confirmed (FIG. 4).

Example 1

The crosslinked hyaluronic acid / gelatin hydrogel prepared as in Preparation Example 1 was placed in an aqueous albumin solution so that all the albumin solution was absorbed into the hydrogel. The crosslinked hyaluronic acid collagen hydrogel containing albumin was dried for 24 hours.

The albumin-containing crosslinked hyaluronic acid / collagen hydrogel was placed in PBS at 37 ° C. and a predetermined amount of solution was taken at a predetermined time to measure the amount of albumin released from the hyaluronic acid hydrogel using a UV spectrometer. (Figure 3)

Example 2

The crosslinked hyaluronic acid / chitosan hydrogel prepared as in Preparation Example 3 was placed in an aqueous albumin solution so that all the albumin solutions were absorbed into the hydrogel. Albumin-sealed crosslinked hyaluronic acid / collagen hydrogel was dried for 24 hours.

The albumin-containing crosslinked hyaluronic acid / gelatin hydrogel was placed in PBS at 37 ° C. and a predetermined amount of solution was taken at a predetermined time to measure the amount of albumin released from the hyaluronic acid hydrogel using a UV spectrometer (FIG. 3).

Example 3

The crosslinked hyaluronic acid / collagen hydrogel prepared as in Preparation Example 4 was placed in an albumin aqueous solution so that all the albumin solutions were absorbed into the hydrogel. The crosslinked hyaluronic acid collagen hydrogel containing albumin was dried for 24 hours.

The albumin-containing crosslinked hyaluronic acid / gelatin hydrogel was placed in PBS at 37 ° C. and a predetermined amount of solution was taken at a predetermined time to measure the amount of albumin released from the hyaluronic acid hydrogel using a UV spectrometer (FIG. 3).

Comparative Example 1

A 1% w / v hyaluronic acid solution was prepared at room temperature, followed by adding 1 mol% adipic dihydrazide and mixing at room temperature to proceed the crosslinking reaction. The cross-linked hyaluronic acid prepared by using adipic dihydrazide (AD) as a crosslinking agent was sufficiently washed with distilled water and then crosslinked hyaluronic acid was dried in a vacuum dryer and subjected to cytotoxicity experiments in the same manner as in Experimental Example 2. (Figure 2)

Comparative Example 2

As in Preparation Example 1, a 1% w / v hyaluronic acid solution was prepared at room temperature, and then 1 mol% poly (ethylene glycol) diglycidyl ether (PEGDE) was added thereto, followed by crosslinking reaction at room temperature. The cross-linked hyaluronic acid prepared using poly (ethylene glycol) diglycidyl ether as a crosslinking agent was sufficiently washed with distilled water and then cross-linked hyaluronic acid was dried in a vacuum dryer, and cytotoxicity experiments were carried out in the same manner as in Experimental Example 2. (Figure 2)

Cytotoxicity was hardly observed in hyaluronic acid / chitosan, hyaluronic acid / gelatin, hyaluronic acid / collagen crosslinked hydrogels prepared using EDC as shown in FIG. 2. However, hyaluronic acid cross-linked using adipic dihydrazide (AD) and poly (ethylene glycol) diglycidyl ether (PEGDE) as crosslinking agents showed cytotoxicity potential from very low cellular activity compared to control. Therefore, the composition according to the present invention was found to be extremely safe in order to be implanted or injected in vivo to express the pharmaceutical function. That is, in the hyaluronic acid / gelatin, hyaluronic acid / chitosan cross-linked hydrogel prepared using EDC as shown in Figure 2, although the general differentiation and proliferation of cells after 24 hours has been confirmed using AD and PEGDE as a crosslinking agent Hyaluronic acid cross-linked products have been shown to inhibit cell proliferation due to toxicity.

Comparative Example 3

 A 1% w / v hyaluronic acid solution was prepared at room temperature, mixed with an aqueous albumin solution to prepare hyaluronic acid containing albumin, and then dried for 48 hours.

Hyaluronic acid containing albumin was placed in PBS at 37 ° C. and a predetermined amount of solution was taken at a predetermined time to measure the amount of albumin released from the hyaluronic acid hydrogel using a UV spectrometer. (Figure 3)

Comparative Example 4

When preparing the hyaluronic acid / gelatin crosslinked hydrogel, the weight fraction of hyaluronic acid in the crosslinked hydrogel was changed to 5,20,40,80,90% to prepare the crosslinked hydrogel, Drug release from the crosslinked hydrogels was compared and confirmed. (FIG. 4).

As can be seen in Figure 3 the amount of drug released from hyaluronic acid cross-linked hyaluronic acid prepared by the present invention shows a slowly increasing pattern over 12 hours. However, the initial drug burst occurred in the non-crosslinked hyaluronic acid and drug mixture phase. In addition, when the hyaluronic acid / polysaccharide cross-linked hydrogel as shown in Figure 4 when changing the weight fraction of hyaluronic acid in the cross-linked hydrogel was confirmed the effect on drug release. When the weight fraction of hyaluronic acid in the cross-linked hydrogel is 30 to 80%, the drug is slowly released over 12 hours, but when 20% or less, or 90% or more, it was confirmed that the drug release mechanism was not expressed.

As described above, the detailed description of the present invention has been made by the embodiments with reference to the accompanying drawings. However, since the above-described embodiments have only been described with reference to preferred examples of the present invention, the present invention is limited to the above embodiments. It should not be understood that the scope of the present invention is to be understood by the claims and equivalent concepts described below.

Claims (8)

1. Polysaccharides, such as hyaluronic acid, which are used in pharmaceutical formulations or tissue engineering preparations, such as injections, contain protein drugs or anti-inflammatory drugs, and polysaccharides hyaluronic acid and Drug Delivery Composition Consisting of Crosslinked Biocompatible Polymers
2. The method of claim 1,

   Drug delivery composition, characterized in that the molecular weight of the hyaluronic acid is 30,000 Daltons (Da) to 3,000,000 Daltons (Da)
3. The method of claim 1,

   The drug delivery composition,

   Any one selected from the group consisting of carboxyl groups of hyaluronic acid and other biocompatible polymers and biodegradable polymers using EDC (1-Ethyl-3- (3-dimethylaminopropyl) -carbodiimide) in a 0.5 to 10 wt% aqueous solution of hyaluronic acid Hyaluronic acid hydrogels crosslinked by an amide reaction of; And

   Protein drugs or nonsteroidal anti-inflammatory agents; Drug delivery composition characterized in that it contains
4. The method of claim 3, wherein

   The biocompatible polymer is a drug delivery composition, characterized in that selected from the group consisting of methyl cellulose, carboxymethyl cellulose, hydroxy propylmethyl cellulose, alginate, chitosan, gelatin, collagen and mixtures or derivatives thereof
5. The method of claim 3, wherein

   The biodegradable polymer,

   Drug delivery composition, characterized in that it is selected from homopolymers of lactic acid in the range of molecular weights from 6,000 to 100,000, copolymer derivatives of lactic acid and glycolic acid, or mixtures thereof
6. The method of claim 1,

   The protein drug is a drug delivery composition, characterized in that selected from the group consisting of albumin, insulin, erythropoietin, insulin growth factor, platelet-derived growth factor, modified growth factor alpha, modified growth factor beta, bone-forming protein and mixtures thereof
7. The method of claim 1,

   The anti-inflammatory drug is a drug delivery composition, characterized in that it is selected from the group consisting of pyroxicam, meloxycampin, ibuprofen, metoprofen, diclofenac, indomethacin, tetracycline, monocycline, doxycycline and mixtures thereof
8. The method of claim 3, wherein

   Drug delivery composition, characterized in that the content of hyaluronic acid in the cross-linked hyaluronic acid hydrogel is 30 to 80% by weight

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