KR102580370B1 - Adhesion barrier containing natural-polymer with biocompatibility and DNA fragment mixture, and process for producing the same - Google Patents

Abstract

The present invention relates to an anti-adhesion agent containing a mixture of biocompatible natural polymers and DNA fragments, and a method for producing the same. The anti-adhesion agent of the present invention not only exhibits excellent biocompatibility and has an excellent anti-adhesion effect, but also heals by reducing inflammatory reactions. It has the effect of increasing the healing speed of tissue damage caused by surgery by improving the speed.

Description

Adhesion barrier containing natural-polymer with biocompatibility and DNA fragment mixture, and process for producing the same}

The present invention relates to an anti-adhesion agent containing a mixture of biocompatible natural polymers and DNA fragments and a method for producing the same. More specifically, it relates to an anti-adhesion agent that improves tissue regeneration in areas damaged by surgery while having an inhibitory effect on tissue adhesion that frequently occurs after surgery.

Adhesion refers to a phenomenon in which tissues or organs of the human body that should be separated from each other form fibrous tissue due to a wound and do not separate from each other (Kwon Y.H., et al., 2004). It can be caused by inflammation, wounds, friction, surgery, etc. During the healing process of wounds such as wounds, it may occur due to excessive production of fibrous tissue or blood leakage and coagulation.

This adhesion phenomenon is a common complication after surgical operations, and serious aftereffects can occur when organs or tissues around the surgical area adhere to each other during the postoperative recovery process. For example, in the case of abdominal surgery, intestinal dysfunction, intestinal obstruction, and chronic pain may occur, and adhesions after gynecological surgery can cause infertility. According to statistics, the incidence of long-term adhesions after surgery is reported to range from 55 to 93%. Adhesions occur frequently after open surgery, and if symptoms do not improve with non-surgical treatment, surgical treatment should be actively considered. It causes many difficulties for doctors and patients during surgery (Kwon S.W., et al., 2006; Lee Y.W., et al., 2009).

The mechanism of adhesion is organ damage and loss of the epithelial cell surface at the surgical site, which causes an inflammatory response and increases vascular permeability of the wound tissue, producing a large amount of serous blood exudate. This serous exudate contains fibrinogen and plasminogen, which is converted to fibrin by thrombin and causes initial fibrous adhesions. Plasminogen is converted to plasmin by plasminogen activator, which is a special fibrinolytic enzyme that dissolves initial adhesions (Kwon Y.H., et al., 2004). If the fibrin adhesion is not absorbed even after 3 days after surgery, fibroblasts proliferate and combine with other tissues to form adhesions. Around 5 days after surgery, collagen production begins and microvessles containing endothelial cells appear. The hardening of these collagen bundles and the granulation tissue where blood vessels are distributed become elements that constitute adhesion (Lee J.H., et al., 2004).

The adhesion prevention methods currently in use are broadly classified into three categories. First, a method to minimize tissue damage and adhesions caused by foreign substances due to delicate care and unnecessary procedures during surgery, second, a method to suppress inflammatory reactions and adhesion formation through drug treatment based on the adhesion mechanism in the pathophysiological process required for adhesion formation, third. , This is a method of preventing adhesion by blocking contact with surrounding tissues by wrapping or covering the wound area using an anti-adhesion barrier after surgery (Oh A.G., 2013).

A suitable material as an anti-adhesion membrane must be stable and effective, not cause inflammatory or immune reactions, do not require suturing, and must remain active in the blood. In addition, it remains during the healing period of the damaged tissue and acts to prevent adhesion, but is decomposed or absorbed after a certain period of time, so there is no need for separate removal. It must be easy to apply to the wound site through laparoscopic surgery and open surgery, and it must be decomposed and metabolized. Substances emitted through the system must also be harmless to the human body and have excellent biocompatibility.

Anti-adhesion barriers can be broadly divided into two types: intra-peritoneal instillators and adhesion barriers. Intraperitoneal drops can be divided into solution type and gel form, and most adhesion blocking agents are in the form of film and membrane (Kamel R.M., 2010).

Problems with the currently commercialized anti-adhesion membranes in the form of solutions, gels, and films are being presented. Even if anti-adhesion agents in the form of solutions have a high viscosity, they flow out of the body and are difficult to apply accurately to the wound. They often flow into other areas before performing the anti-adhesion function or are decomposed too early, so they often do not perform the anti-adhesion function properly. There is a downside. Anti-adhesion membranes in the form of films and membranes have been clinically proven to be the most effective, but when applied to internal organs, they do not adhere well to the surface of the organ, and even if they do adhere, they are not consistently and accurately positioned at the wound site due to the movement of the organ and the tissue itself. It is reported that the effect of preventing long-term adhesion is insufficient as it is recognized as a foreign substance and clumps together. Gel-type anti-adhesion agents are injectable formulations that are being developed to overcome the disadvantages of film and solution forms. The time required for wound healing varies depending on the severity of the wound, but is generally about 7 days. In order to expect an anti-adhesion effect, the wound area must be helped to regenerate normally for a period of about 7 days, and the tissues adjacent to the wound must be helped to regenerate normally. It must prevent the formation of fibrous tissue and then naturally decompose, absorb, and eliminate it. However, in the case of gel-type anti-adhesion agents, they are dissolved and discharged before the wound heals and do not have enough time to stay in the wound tissue, so they do not properly show the anti-adhesion effect, and non-living substances can cause problems such as foreign body reactions in vivo. There is (Oh A.G., 2013).

The main ingredients used in commercially available adhesion blockers include the natural polymers sodium carboxymethyl cellulose (CMC), sodium hyaluronate (HA), and alginate, and the synthetic polymers polyethylene glycol- There are polymers such as polyethylene glycol-polyproplene glycol (PEG-PPG), poloxamer 188 (PEG-PPG-PEG), and poloxamer 407 (PEG-PPG-PEG) (Park J.K., et al., 2015).

As mentioned above, natural polymers and synthetic polymers are widely used to prevent adhesion, but it is difficult to achieve the desired effect of preventing adhesion. In the case of synthetic polymers, the biocompatibility is significantly lower than that of natural polymers, and since there are no decomposition enzymes in the body, there is a possibility that they may remain in the body unnecessarily, causing side effects such as unexpected tissue reactions. In the case of natural polymers, while they have excellent biocompatibility, their absorption period in the body is short, so it is difficult to expect excellent anti-adhesion efficacy. In addition, the problem of slowing wound recovery in the area where the anti-adhesion agent was applied has been identified, so a method to overcome this problem is needed.

Accordingly, the present inventors were able to complete the present invention as a result of researching an anti-adhesion agent that has excellent biocompatibility and excellent anti-adhesion efficacy and can improve tissue regeneration at the surgical site.

As a prior art, Korean Patent No. 1452041 discloses an anti-adhesion material containing hyaluronic acid, carboxymethyl cellulose and alginate, and Korean Patent No. 986603 discloses a mixture of DNA fragments for the purpose of improving the treatment of wounds. Although it has been disclosed that it can be used in pharmaceuticals, there has been no disclosure about an anti-adhesion agent mixed with a mixture of biocompatible natural polymers and DNA fragments.

Korean Patent No. 1452041, Anti-adhesion material and anti-adhesion method, registered on October 10, 2014. Korean Patent No. 986603, DNA polymer fragment mixture isolated from fish semen or eggs and method for manufacturing the same, registered on October 4, 2010.

Kamel R.M., et al., Prevention of postoperative peritoneal adhesion, Eur. J. Obstet. & Gynecol. and Rep. Biol., 150, 111, 2010. Kwon S.W., et al., Anti-adhesive effect of poloxamer/alginate/CaCl2 mixture in the rat model, J. Korean Surg. Soc., 71, 280-287, 2006. Kwon Y.H., et al., Prevention of postsurgical adhesions with a mixed gel of sodium carboxymethyl cellulose and gellan gum in the rat, J. Kor. Pham. Sci., 34, 477-482, 2004. Lee J.H., et al., Effect of hyaluronic acid on prevention of adhesion in rats, Korean J. Vet. Res., 44, 663-668, 2004. Lee Y.W., et al., Efficacy and safety of the electrospun nanofibrous adhesion barrier for laparoscopic surgery in a rabbit model, J. Korean Surg. Soc., 76, 73-80, 2009. Oh A.G., Trend of anti-adhesion adjuvant -Review, Biomater. Res., 17, 138-145, 2013. Park J.K., et al., Thermosensitive chitosan-based hydrogel with growth factor as adhesion barrier, Polymer (Korea), 39, 480-486, 2015.

The purpose of the present invention is to provide an anti-adhesion agent containing a mixture of biocompatible natural polymers and DNA fragments and a method for producing the same.

The present invention relates to an anti-adhesion agent containing a mixture of biocompatible natural polymers and DNA fragments.

The DNA fragment mixture may have a molecular weight of 50 kDa to 3,000 kDa.

The biocompatible natural polymer may be 0.3% by weight to 10% by weight based on the total weight.

The DNA fragment mixture may be 0.3% to 5% by weight based on the total weight.

The weight ratio of the biocompatible natural polymer and DNA fragment mixture may be 0.5:1 to 10:1.

The biocompatible natural polymers include alginic acid, pectin, carrageenin, gellan gum, hyaluronic acid, carboxymethyl cellulose, and dextran. It may be one or more types of polymers selected from the group consisting of.

The biocompatible natural polymer may be hyaluronic acid.

The DNA fragment mixture may be isolated from fish testes or semen.

In addition, the present invention includes the steps of dissolving a mixture of biocompatible natural polymers and DNA fragments in an aqueous solvent to produce a mixed aqueous solution; and checking or controlling the viscosity of the mixed aqueous solution.

Hereinafter, the present invention will be described in detail.

The adhesion may refer to a phenomenon in which separated skin or membranes stick to each other. When tissue damage occurs after a surgical operation or due to inflammation, foreign body, bleeding, infection, wound, friction, chemical treatment, etc., blood leaks out and coagulates during the healing process of the wound, resulting in an adhesion phenomenon where abnormal adhesion occurs with surrounding tissue. do. These adhesions occur especially frequently after surgery, and when adhesions occur in the pelvis, they can cause chronic pain and sexual dysfunction. Adhesions caused by the formation of scars after thyroid removal surgery can cause chest pain and decreased swallowing ability. Side effects occur, and when adhesions occur due to spinal surgery, nerves are compressed, causing severe pain, and intrauterine adhesions are known to cause infertility, amenorrhea, and habitual miscarriage.

The anti-adhesion agent of the present invention uses a mixture of biocompatible natural polymers and DNA fragments to improve the cell support role and wound healing environment to promote tissue regeneration.

The biocompatible natural polymer may be included in an amount of 0.3% to 10% by weight, preferably 0.5% to 5% by weight, based on the total weight of the composition. If the biocompatible natural polymer is included in less than 0.3% by weight, the anti-adhesion effect may be limited, and if it is included in more than 10% by weight, manufacturing problems may occur due to viscosity.

The biocompatible natural polymers include alginic acid, pectin, carrageenin, gellan gum, hyaluronic acid, carboxymethyl cellulose, and dextran. It may be one or more types of polymers selected from the group consisting of. Preferably it may be hyaluronic acid.

The hyaluronic acid is a natural acidic mucopolysaccharide in the form of a straight chain in which β-D-N-acetylglucosamine and β-D-glucuronic acid are alternately bonded to β-1, 3 and β-1, 4, and the molecular weight is 50,000 by an adjustment method. It is a polymer compound ranging from 800,000 to 800,000. It has no species or organ specificity and has excellent biocompatibility and very high viscoelasticity even when implanted or injected into the living body. Due to these characteristics, hyaluronic acid can be used for various purposes such as inserts for arthritis treatment, anti-adhesion gel/film, drug carrier, and cosmetic supplement.

The hyaluronic acid may have a molecular weight of 500 kDa to 8,000 kDa, preferably 500 kDa to 3,000 kDa. At this time, if the molecular weight is less than 500 kDa, the residence time in the body is short and the anti-adhesion effect is weak, and if the molecular weight is more than 8,000 kDa, the viscosity increases and problems in the manufacturing process such as fluid transfer and filtration may occur, which is not desirable.

The DNA fragment mixture refers to DNA corresponding to a biopolymer consisting of phosphoric acid, four types of bases, and deoxyribose, which exists mixed in the form of fragments with reduced molecular weight.

The DNA fragment mixture may be included in an amount of 0.3% to 5% by weight, preferably 0.5% to 5% by weight, based on the total weight of the composition. If the DNA fragment mixture is less than 0.3% by weight, it is difficult to expect a practical regeneration effect, and if it is more than 5% by weight, problems may occur in the manufacturing process of the DNA fragment mixture, which is not desirable.

The DNA fragment mixture may have a molecular weight of 50 kDa to 3,000 kDa, preferably 50 kDa to 1,500 kDa. At this time, if the molecular weight of the DNA fragment mixture is less than 50 kDa or more than 3,000 kDa, it is difficult to satisfy tissue regeneration as an anti-adhesion agent.

When hyaluronic acid is used alone as the anti-adhesion agent, the residence time in the body is relatively short due to the biodegradability of hyaluronic acid due to its excellent biocompatibility, so the anti-adhesion effect is weak, and due to its high water solubility, it diffuses and flows from the surface of the wound in a short period of time. A problem appears. The anti-adhesion agent comprising a mixture of biocompatible polymer and DNA fragment of the present invention solves the above problems and exhibits excellent biocompatibility and tissue regeneration.

The ratio of the biocompatible natural polymer and DNA fragment mixture may be 0.5:1 to 10:1 by weight. Preferably, the weight ratio may be 1.5:1 to 3:1. When mixed at this weight ratio, the viscosity, adhesion, and applicability as an anti-adhesion agent were excellent.

Flowing can be controlled by mixing the biocompatible natural polymer and DNA fragment mixture, and a viscosity regulator can be added to control viscosity.

There is no problem in using the viscosity modifier if its safety has been confirmed to have no side effects when injected into the human body.

The viscosity modifier includes glucomannan, guar gum, locust bean gum, xanthan gum, glucose, carboxymethylated starch, mannose, It may be galactose, arabinose, fucose, ribose, fructose, etc.

Additionally, the present invention provides an anti-adhesion agent containing a mixture of biocompatible natural polymers and DNA fragments and having an absolute viscosity of 100 cps to 5,000 cps. In general, 500cps is considered to be the viscosity of engine oil, and 5,000cps is considered to be the viscosity of corn syrup.

If the absolute viscosity is less than 100 cps, it is difficult to provide sufficient adhesion prevention and adhesion due to low viscosity, and if it exceeds 5,000 cps, the viscosity becomes very large, making stirring and polymerization difficult, and it is not desirable because it cannot be properly distributed to the desired area in the tissue. . Considering anti-adhesion properties, adhesion, flowability, and ease of use, the viscosity of the anti-adhesion agent is preferably 100 cps to 2,000 cps, and most preferably 100 cps to 1,500 cps.

The formulation of the anti-adhesion agent can be divided into solution formulation, gel formulation, and film formulation.

When the solution formulation is used by spraying, it is preferable to have a particle size of less than 200㎛ to enable spraying, and in the case of simple application, it is preferable to have particles of less than 1,000㎛.

The film formulation often needs to be removed and reapplied after the procedure. Therefore, when formulating it into a film formulation, it is desirable that the anti-adhesion agent not only has excellent initial adhesion but also excellent re-adhesion, and the scope of the present invention is not limited to this, but can be applied to various types of preparations.

In addition, the present invention includes the steps of dissolving a mixture of biocompatible natural polymers and DNA fragments in an aqueous solvent to produce a mixed aqueous solution; and checking or controlling the viscosity of the mixed aqueous solution.

In the step of generating the mixed aqueous solution, water, physiological saline solution, distilled water for injection, etc. may be used as an aqueous solvent.

The step of checking or adjusting the viscosity of the mixed aqueous solution is a step of checking whether the viscosity of the mixed aqueous solution is within a certain range, that is, preferably within the range of 100 cps to 1,500 cps, or if it is outside this range, the viscosity This is a step of adjusting the viscosity of the mixed aqueous solution within the range of 100 cps to 1,500 cps by additionally adding a regulator.

The present invention relates to an anti-adhesion agent containing a mixture of biocompatible natural polymers and DNA fragments and a method for producing the same. The anti-adhesion agent according to the present invention not only exhibits excellent biocompatibility and has an excellent anti-adhesion effect, but also improves tissue regeneration and is suitable for surgery. It has the effect of increasing the healing speed of tissue damage caused by .

Figure 1 shows a rat that underwent laminectomy to confirm the anti-adhesion effect.
Figure 2 shows the results of evaluating the degree of adhesion according to treatment with an anti-adhesion agent.
Figure 3 shows the results of confirming the degree of inflammation following treatment with an anti-adhesion agent. (A) shows the expression rate of TNF-α and (B) shows the expression rate of IL-6.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the content introduced herein is provided to be thorough and complete, and to fully convey the spirit of the present invention to those skilled in the art.

<Experimental Example 1. Confirmation of anti-adhesion effect according to DNA fragment mixture concentration>

Experimental Example 1-1. Preparation of anti-adhesion agent composition according to DNA fragment mixture concentration

In order to confirm the concentration range of the DNA fragment mixture that can be used when preparing the anti-adhesion agent of the present invention, a composition was prepared by varying the concentration of the DNA fragment mixture. At this time, hyaluronic acid (HA) was used as a biocompatible natural polymer, and if the concentration of hyaluronic acid used is less than 0.3% by weight based on the total weight of the composition, there is almost no anti-adhesion effect, and if it exceeds 10% by weight, it has little effect on preventing adhesion. When included, there was a manufacturing problem due to viscosity, so the concentration was fixed at 1.5% by weight, which is within the range of 0.3% by weight to 10% by weight.

A mixture of hyaluronic acid (HA) and DNA fragments was dissolved in physiological saline to obtain the concentration shown in Table 1 below to create a mixed aqueous solution, and the viscosity was measured, and the results are shown in Table 2.

composition Hyaluronic acid (% by weight) DNA fragment mixture (% by weight) Experimental Example 1-1 1.5 0 Experimental Example 1-2 1.5 0.1 Experimental Example 1-3 1.5 0.3 Experimental Example 1-4 1.5 0.5 Experimental Example 1-5 1.5 One Experimental Example 1-6 1.5 2 Experimental Example 1-7 1.5 4 Experimental Example 1-8 1.5 5

composition Viscosity of mixed aqueous solution
(cps) Experimental Example 1-1 1946 Experimental Example 1-2 1712 Experimental Example 1-3 1487 Experimental Example 1-4 1224 Experimental Example 1-5 831 Experimental Example 1-6 503 Experimental Example 1-7 185 Experimental Example 1-8 103

As shown in Table 2, it was found that as the concentration of the DNA fragment mixture increased, the viscosity of the mixed aqueous solution of hyaluronic acid and DNA fragment mixture decreased. In addition, through the above viscosity and visual inspection, it was determined that it would be suitable for use as an anti-adhesion agent when the viscosity was 100 cps to 2,000 cps.

Experimental Example 1-2. Confirmation of anti-adhesion effect of anti-adhesion agent composition according to concentration of DNA fragment mixture

A laminectomy was performed to confirm the effect of preventing adhesion. Laminectomy is a surgical method for degenerative spinal diseases, and is known to cause many adhesions despite surgical decompression mainly centered on the nerve roots of the lumbar spine.

To perform laminectomy, 10-week-old S/D (Sprague Dawley) male white rats were used. All mice consumed solid food and water freely and were raised in a laboratory environment where the humidity was maintained at 60% at 22-24°C and the day and night cycle was adjusted to 12-hour intervals.

Rats bred as shown in Figure 1 were general anesthetized by intraperitoneal injection of Zoletil 50 ^® , then laminectomy was performed on the 4th and 5th lumbar vertebrae to expose the dura, and then hemostasis was performed. Experimental Example 1-3, Experimental Example 1-4, Experimental Example 1-8 and as a positive control group, currently sold anti-adhesion agents Guardix-sol (Control 1) and Mediclore (Control 2) were applied. For wound closure, the muscle layer was sutured and then the skin was sutured. In the untreated group, laminectomy was performed on the same area and wound closure was performed immediately after hemostasis. At this time, the number of experimental animals was 10 each in the experimental group (Experimental Example 1-3, Experimental Example 1-4, Experimental Example 1-8) and control group (Control Group 1, Control Group 2) for each week, and 5 animals in the untreated group. .

Experimental animals were sacrificed at 2, 4, and 8 weeks after surgery to check macroscopic and microscopic findings to evaluate the amount of scar tissue and degree of adhesion. The scores range from 0 to 3 according to each symptom in Table 3 below. They were evaluated separately, and the results are shown in Table 4.

score amount of scar tissue Adhesion 0 doesn't exist No attachment One small amount of scarring Slight adhesions to the dura: easily removed 2 Moderate amount of scarring Moderate adhesion: requires force to come off 3 large amount of scarring Sticky degree of adhesion: forced separation by sharp object

composition cicatricial Adhesion 2 weeks 4 weeks 8 weeks 2 weeks 4 weeks 8 weeks Untreated group 2.0 2.2 2.2 1.8 2.2 2.4 Experimental Example 1-3 0.2 0.5 0.8 0 0.4 0.7 Experimental Example 1-4 0.1 0.5 0.6 0 0.3 0.5 Experimental Example 1-8 0.3 0.8 0.9 0 0.3 0.6 Control group 1 (Gaddix) 0.6 1.1 1.3 0.4 0.9 1.1 Control group 2 (Mediclo) 0.7 1.3 1.7 0.5 1.1 1.3

As shown in Table 4, it can be seen that the anti-adhesion agent composition containing the DNA fragment mixture has a higher anti-adhesion effect than Control 1 and Control 2, which do not contain the DNA fragment mixture.

In addition, it can be confirmed that the anti-adhesion agent composition containing the DNA fragment mixture increases the anti-adhesion effect and at the same time accelerates healing at the surgical site.

<Example 1. Preparation of anti-adhesion agent>

An anti-adhesion agent containing a mixture of biocompatible natural polymer and DNA fragment of the present invention was prepared through the same process as Experimental Example 1-1.

At this time, the weight percent of the hyaluronic acid and DNA fragment mixture was prepared as the weight percent corresponding to the examples in Table 5 below.

<Comparative Example 1. Preparation of comparative anti-adhesion agent>

A comparative anti-adhesion agent containing a mixture of biocompatible natural polymers and DNA fragments was prepared under the weight percent conditions corresponding to the comparative examples in Table 5 below. The manufacturing method was performed in the same manner as in Experimental Example 1-1.

composition Content (% by weight) Content ratio Hyaluronic acid DNA fragment mixture Hyaluronic acid DNA fragment mixture Example 1-1 0.3 0.6 0.5 One Example 1-2 0.45 0.3 1.5 One Example 1-3 0.6 0.3 2 One Example 1-4 0.9 0.3 3 One Examples 1-5 1.5 0.3 5 One Example 1-6 3 0.3 10 One Example 1-7 1.5 0.5 3 One Examples 1-8 10 5 2 One Example 1-9 10 3.3 3 One Examples 1-10 10 2 5 One Example 1-11 10 One 10 One Comparative Example 1-1 0 One 0 One Comparative Example 1-2 10 0.5 20 One Comparative Example 1-3 0.3 One 0.3 One Comparative Example 1-4 10 6.7 1.5 One

< Example 2. Check the anti-adhesion effect>

In order to confirm the anti-adhesion effect of the anti-adhesion agent of Example 1 and Comparative Example 1, the same experiment as the experimental method of Experimental Example 1-2 was performed, and the degree of adhesion was measured 8 weeks after treatment with the anti-adhesion agent. As a positive control group, currently sold anti-adhesion agents Guardix-sol (Control 1) and Mediclore (Control 2) were applied. The results of the degree of adhesion are shown in Table 6 and Figure 2.

composition Content (% by weight) Degree of adhesion Hyaluronic acid DNA fragment mixture Untreated group - - 2.8±0.2 Example 1-1 0.3 0.6 1.2±0.6 Example 1-2 0.45 0.3 0.9±0.4 Example 1-3 0.6 0.3 0.8±0.4 Example 1-4 0.9 0.3 0.7±0.2 Examples 1-5 1.5 0.3 1.0±0.4 Example 1-6 3 0.3 1.1±0.6 Example 1-7 1.5 0.5 0.5±0.4 Examples 1-8 10 5 0.9±0.6 Example 1-9 10 3.3 0.7±0.6 Examples 1-10 10 2 0.9±0.8 Example 1-11 10 One 1.2±0.4 Comparative Example 1-1 0 One 2.4±0.4 Comparative Example 1-2 10 0.5 1.5±0.8 Comparative Example 1-3 0.3 One 1.6±1.0 Comparative Example 1-4 10 6.7 1.9±0.8 Control group 1 (Gaddix) - - 1.6±1.0 Control group 2 (Mediclo) - - 1.8±0.8

As shown in Table 6, it was confirmed that treatment with the anti-adhesion agent of Example 1 had the best anti-adhesion effect compared to the anti-adhesion agent of Comparative Example 1 or the control group.

<Experimental Example 2. Confirmation of wound healing degree>

Adhesions are caused by organ damage and loss of the epithelial cell surface at the surgical site, resulting in an inflammatory response and increased vascular permeability of the wound tissue, producing a large amount of serous blood exudate. Fibrinogen in this serous exudate is converted to fibrin by thrombin and causes initial fibrous adhesions. Therefore, a reduction in the inflammatory response occurring at the site of organ damage or surgery is associated with a decrease in organ adhesion. Accordingly, the degree of inflammatory response following treatment with the anti-adhesion agent of the present invention was observed.

After inducing adhesion in Example 2, blood was collected from mice 2 weeks after treatment with an anti-adhesion agent, and the expression level of inflammatory cytokines in the collected blood was measured. At this time, TNF-α and IL-6 were measured as inflammatory cytokines, and the expression of each cytokine was confirmed using an ELISA (enzyme-linked immunosorbent assay) kit, and the results are shown in Table 7 and Figure 3.

composition Content (% by weight) TNF-α IL-6 Hyaluronic acid DNA fragment mixture Untreated group - - 100 100 Example 1-1 0.3 0.6 74.2±8.4 72.1±7.8 Example 1-2 0.45 0.3 67.9±10.1 65.1±9.1 Example 1-3 0.6 0.3 60.6±7.8 59.4±8.4 Example 1-4 0.9 0.3 54.4±6.7 57.1±6.8 Examples 1-5 1.5 0.3 51.1±9.4 53.9±7.5 Example 1-6 3 0.3 54.2±8.4 56.1±7.8 Example 1-7 1.5 0.5 49.7±7.7 52.7±10.8 Examples 1-8 10 5 62.1±12.6 63.9±9.1 Example 1-9 10 3.3 67.1±10.4 68.2±9.4 Examples 1-10 10 2 72.1±11.9 74.2±8.9 Example 1-11 10 One 78.7±12.8 80.6±10.1 Comparative Example 1-1 0 One 91.7±7.5 92.8±6.7 Comparative Example 1-2 10 0.5 88.8±10.9 85.1±9.9 Comparative Example 1-3 0.3 One 85.7±9.4 84.1±9.7 Comparative Example 1-4 10 6.7 86.1±7.3 87.5±6.7 Control group 1 (Gaddix) - - 90.4±9.1 91.7±6.2 Control group 2 (Mediclo) - - 92.1±7.7 91.0±8.4

As shown in Table 7, it can be seen that the inflammation level is reduced when treated with the anti-adhesion agent of Example 1 compared to the anti-adhesion agent of Comparative Example 1 or the control group.

Through this, it can be predicted that the anti-adhesion agent of Example 1 of the present invention increases the anti-adhesion effect by reducing the inflammatory response of the wound and shortening the wound healing time.

Claims (9)

An anti-adhesion agent containing a mixture of hyaluronic acid and DNA fragments, wherein the weight ratio of the mixture of hyaluronic acid and DNA fragments is 3:1. According to paragraph 1,
The DNA fragment mixture is an anti-adhesion agent, characterized in that the molecular weight is 50 kDa to 3,000 kDa. According to paragraph 1,
An anti-adhesion agent, characterized in that the hyaluronic acid is 0.3% by weight to 10% by weight based on the total weight. According to paragraph 1,
An anti-adhesion agent, characterized in that the DNA fragment mixture is 0.3% by weight to 5% by weight based on the total weight. delete delete delete According to paragraph 1,
An anti-adhesion agent, characterized in that the DNA fragment mixture is isolated from testes or semen of fish. As a method for producing the anti-adhesion agent according to claim 1,
Dissolving a mixture of hyaluronic acid and DNA fragments in an aqueous solvent to produce a mixed aqueous solution; and
A method for producing an anti-adhesion agent, comprising the step of checking or controlling the viscosity of the mixed aqueous solution.