WO2015088275A1 - Biodegradable medical adhesive or sealant composition - Google Patents

Biodegradable medical adhesive or sealant composition Download PDF

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Publication number
WO2015088275A1
WO2015088275A1 PCT/KR2014/012265 KR2014012265W WO2015088275A1 WO 2015088275 A1 WO2015088275 A1 WO 2015088275A1 KR 2014012265 W KR2014012265 W KR 2014012265W WO 2015088275 A1 WO2015088275 A1 WO 2015088275A1
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Prior art keywords
oxidized
component
composition
adhesion
hyaluronic acid
Prior art date
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PCT/KR2014/012265
Other languages
French (fr)
Korean (ko)
Inventor
이은혜
맹진희
김근수
이돈행
박영환
Original Assignee
재단법인 유타 인하 디디에스 및 신의료기술개발 공동연구소
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Priority claimed from KR1020140089173A external-priority patent/KR101664444B1/en
Application filed by 재단법인 유타 인하 디디에스 및 신의료기술개발 공동연구소 filed Critical 재단법인 유타 인하 디디에스 및 신의료기술개발 공동연구소
Priority to AU2014360958A priority Critical patent/AU2014360958B2/en
Priority to RU2016128376A priority patent/RU2657836C1/en
Priority to CN201480067775.9A priority patent/CN106061518B/en
Priority to CA2933271A priority patent/CA2933271C/en
Priority to US15/102,406 priority patent/US10105465B2/en
Priority to JP2016538741A priority patent/JP6207745B2/en
Priority to EP14870365.5A priority patent/EP3081236B1/en
Priority to MX2016007688A priority patent/MX366143B/en
Publication of WO2015088275A1 publication Critical patent/WO2015088275A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
    • A61L24/08—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/001—Use of materials characterised by their function or physical properties
    • A61L24/0031—Hydrogels or hydrocolloids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/001—Use of materials characterised by their function or physical properties
    • A61L24/0042—Materials resorbable by the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0023—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0061—Use of materials characterised by their function or physical properties
    • A61L26/008—Hydrogels or hydrocolloids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0061—Use of materials characterised by their function or physical properties
    • A61L26/009—Materials resorbable by the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/20—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/52—Hydrogels or hydrocolloids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/58—Materials at least partially resorbable by the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/04—Macromolecular materials
    • A61L31/042—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/145—Hydrogels or hydrocolloids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/148—Materials at least partially resorbable by the body

Definitions

  • the present invention relates to a biodegradable medical adhesive or sealant composition
  • a biodegradable medical adhesive or sealant composition comprising an oxidized glycosaminoglycan having a formyl group and a polyamine.
  • Bioadhesives and sealants are used to seal or apply tissues during surgery, or to be used as anti-bleeding agents (hemostasis), body fluids and blood blockers. Bioadhesives and sealants require biocompatibility as they come into contact with the skin. It must be absent, biodegradable, and not interfere with the healing of living organisms.
  • cyanoacrylates include cyanoacrylates, fibrin glues, gelatin glues, and polyurethanes. Closure Medi Cal of the United States commercialized a medical tissue adhesive of octylcyanoacrylate called Dermabond in 1997. It was approved by the US FDA in 1998 after it was approved by the European Community in August. However, cyanoacrylate-based adhesives may interfere with wound healing because the solids lack rigidity and rigidity, and are difficult to decompose in vivo, and thus are easily encapsulated and become foreign matters. there was. In addition, since fibrin glue has a considerably low adhesive force, the generated fibrin fish may drop from the tissue, and since the blood product is a blood product, there is a problem of a viral infection.
  • Korean Patent Publication No. 10-2009-0083484 discloses a medical two-component adhesive comprising an aldehyde-ized textan powder and an ⁇ -poly—L-lysine powder prepared by mechanical grinding after lyophilization.
  • LYDEX, Rydex is disclosed.
  • the two-component adhesive is characterized in that it is a powdered medical adhesive, it takes a relatively long time to decompose the gel, there is a problem that does not obtain a satisfactory effect in the water absorption.
  • new products with improved characteristics in terms of decomposition time, adhesion and water absorption There is an increasing demand for medical adhesives.
  • numerous citations and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.
  • the present inventors can perform sufficient adhesion, coating and hemostasis even in the part of the body where body fluid and blood are present, and can absorb a large amount of water, especially compared to other biodegradable polymers, and can decompose itself in the body. And research efforts were made to develop sealants. As a result, the present invention is confirmed that the use of a formyl group and oxidized glycosaminoglycan and polyamine together can effectively bond, layer, apply, prevent adhesion, wound coating and hemostasis of living tissues. It was completed.
  • an object of the present invention is to provide a medical adhesive or sealant (or a medical adhesive or sealant composition).
  • Another object of the present invention is to provide a method for adhesion, layer deposition, application, adhesion prevention, wound coating or hemostasis of living tissue.
  • the invention provides a biodegradable medical adhesive or sealant (or medical adhesive or sealant composition) comprising:
  • a first component comprising an oxidized glycosaminoglycan in which a formyl group is introduced and oxidized
  • a second component comprising a polyamine having two or more amino groups, wherein the pH in the aqueous solution state of the second component is 8.5 to 11.0.
  • the invention provides a biodegradable medical adhesive or sealant (or medical adhesive or sealant composition) comprising:
  • a second component comprising a polyamine having two or more amino groups, the pH in the aqueous solution state of the second component is 8.5 to 11.0,
  • the present invention is the step of applying the biodegradable medical adhesive or sealant (or medical adhesive or sealant composition) to a biological tissue that requires adhesion, layering, application, adhesion prevention, wound coating or hemostasis It provides a method of adhesion, layer deposition, coating, adhesion prevention, wound coating or hemostasis of a biological tissue comprising a.
  • the present inventors can perform sufficient adhesion, coating and hemostasis even in the part of the body where body fluids and blood are present, and in particular, can absorb a large amount of water compared to other biodegradable polymers and can decompose itself in the body. And research efforts were made to develop sealants.
  • the composition of the present invention showed an improved effect on gel formation time, adhesion and water absorption than the conventional medical two-component adhesive (LYDEX) (see Tables 5 to 7), In addition, it showed a better effect than the conventional hemostatic agent (Ar i sta TM AH) (see Fig. 12).
  • the composition of the present invention comprises oxidized glycosaminoglycan as the first component.
  • oxidized glycosaminoglycan means that a formyl group (-CH0) is introduced and oxidized to glycosaminoglycan.
  • the glycosamino glycan is distinguished from glucan, which is a 0-glycoside bond of a monosaccharide, in that it is a polysaccharide having a repeating structure of a disaccharide including nucleosamine.
  • Introduction of such a formyl group can be performed by a periodic acid oxidation method.
  • oxidized glycosaminoglyco introduced with the appropriate number (eg, 0.01 to 0.95) formyl groups per anhydrous glucose unit (sugar moiety) by oxidation with glycosaminoglycanol periodic acid or periodicate You can get a space.
  • Oxidation degree of this glycosaminoglycan Calculated according to, having a value from 10 to 99.5%. Oxidized glycosaminoglycans having such a degree of oxidation, when used in combination with the second component, can quickly absorb the blood and body fluids in the body and gelate quickly.
  • the oxidation degree of the oxidized glycosaminoglycan is
  • the oxidation degree of the oxidized hyaluronic acid may be 10-20%.
  • Oxidation degree measurement of the glycosaminoglycan can be carried out by NaOH scoring method. For example, 17.5 g of hydroxylamine hydrochloride and 0.05% methyl orange 6 are mixed in 994 distilled water to form a solution of 0.25 ⁇ / ⁇ hydroxylamine hydrochloride, titrated ⁇ to 4, and then Dissolve 0.1 g of minoglycan in 25 ⁇ of the above solution, titrate it again to pH 4 with 0.1 ⁇ / i sodium hydroxide, , , ⁇ , , ,
  • the oxidized glycosaminoglycan has 0.01 to 0.95 formyl groups per anhydrous glucose unit (sugar residues).
  • the oxidized glycosaminoglycan is composed of oxidized hyaluronic acid, oxidized chondroitin sulfate, oxidized chondroitin, dermatan sulphate, oxidized heparan sulphate, heparin oxidized and keratan oxidized. Is selected from.
  • the gelling ability of the adhesive / sealant composition by using a glycosaminoglycan having a specific molecular weight, the gelling ability of the adhesive / sealant composition, the gelation state retention time, the elasticity of the gel, and the like can be appropriately controlled.
  • the glycosaminoglycan used to obtain the oxidized glycosaminoglycan has a molecular weight of 1,000 to 5 million.
  • the glycosaminoglycan used to obtain the oxidized glycosaminoglycan is 10,000 to 4 million, 50,000 to 350, 100,000 to 350, 100,000 to 3 million, 100,000 to 2.5 million, 100,000 To 2 million, or 100,000 to 1.6 million molecular weight.
  • the first component is 100,000 to
  • Oxidized hyaluronic acid with a molecular weight of 2 million is included. According to one specific example, the molecular weight of the oxidized hyaluronic acid is 100,000 to 1.6 million.
  • the first component includes two or more kinds of oxidized glycosaminoglycans.
  • the weight ratio of these oxidized glycosaminoglycans is 1: 0.5-5. According to one specific example, the weight ratio of the oxidized glycosaminoglycan is 1: 0.5-4, and in another specific example 1: 0.5.
  • the two or more kinds of oxidation Glycosaminoglycans are oxidized hyaluronic acid and oxidized chondroitin sulfate.
  • the weight ratio of the hyaluronic acid oxide and chondroitin oxide is 1: 0.5-5, 1: 0.5-4, 1: 0.5-3.5, 1: 0.5-3, 1: 0.5-2.5, 1: 0.5-2, 1: 0.5 -1.5 or 1: 0.8-
  • the oxidation degree of the oxidized hyaluronic acid is 10-.
  • the oxidation degree of the oxidized hyaluronic acid is 12-40%, in another specific example 12-383 ⁇ 4>, and in another specific example 13-3.
  • the oxidation degree of the oxidized chondroitin sulfate is 10-55%.
  • the oxidation degree of the chondroitin sulfate is 10-50%, in another specific example 10-45%, in another specific example 10-40%, in another specific example 10 -35%.
  • the first component is in powder, liquid or solid (eg pellet form).
  • the powdery first component may be obtained by drying (eg, spray drying, lyophilization, etc.) of an oxidized glycosaminoglycan-containing solution followed by grinding (eg, mechanical grinding).
  • composition of the present invention further comprises a second component comprising a polyamine having two or more amino groups in addition to the first component as the active component.
  • the second component exhibits a pH of 8.5-11.0 in aqueous solution. As confirmed in the following examples, when the pH in the aqueous solution state of the second component is 8.5-11.0, gelation may be performed within a few seconds (see FIG. 7).
  • the second component is 9.0- in an aqueous solution state.
  • the pleamine may further have a secondary and / or tertiary amino group.
  • the second component comprising the polyamine is in powder, liquid or solid (eg, pellets).
  • the powdery second component can be obtained by drying and grinding the polyamine-comprising solution.
  • the polyamine-containing solution may further include a pH adjusting agent so that the pH range in the aqueous solution state of the second component is 8.5-11.0.
  • the pH regulator include monovalent or polyvalent carboxylic acid compounds such as acetic acid, citric acid, succinic acid, glutaric acid, malic acid, fumaric acid and maleic acid, or anhydrides thereof.
  • the polyamine is polylysine, chitosan, albumin, putrescine (putrescine), cadaverine (cadaver ine), spermidine, spermine (spermine), protamine and PEI (Polyethylenimine) is selected from the group consisting of.
  • the polyamine has a molecular weight of 100 or more.
  • the molecular weight of the polyamine may be 1,000 to 200,000.
  • the polyamine is poly-L-lysine.
  • the poly-L-lysine may be ⁇ -poly-L-lysine produced using microorganisms (eg, Straptomyces abulus) or enzymes.
  • the second component may further comprise a ⁇ regulator in addition to the polyamine.
  • the composition of the present invention may further comprise a drug.
  • the drug may be included in the second component.
  • the drug is
  • It may have one or more amine groups, and examples of such drugs include anthracycline-based drugs, gemcitabine, vancomycin, polymycin, methotrexate, protein drugs and peptide drugs.
  • anthracycline-based drugs gemcitabine, vancomycin, polymycin, methotrexate, protein drugs and peptide drugs.
  • the amine group of the drug may also act as the formyl group of the first component, and the three components may form a gel together.
  • the composition of the present invention may be formulated in various forms, for example, a first component which is in powder form, liquid form or solid form (for example, pellet form), and is in powder form, liquid form or solid form. Combinations of second components.
  • the composition of the present invention comprises the first component and the second component in a weight ratio of 0.5-10: 1.
  • the first component and the second component are included in a weight ratio of 0.5-8: 1, in another specific example, a weight ratio of 0.5-6: 1, and in another specific example, 0.5-4: 1 Weight ratio, in another specific example a weight ratio of 0.5-3: 1, in another specific example, a weight ratio of 0.5-2: 1, in another specific example, a weight ratio of 0.5-1.5: 1, and in another specific example, 0.8-1.5: Weight ratio of 1, and in another specific example, weight ratio of 0.8-1.2: 1.
  • the present invention by controlling the ratio of the formyl group of the first component and the amino group of the second component it is possible to control the gel formation time, the decomposition time of the resulting gel and the like.
  • the molar ratio of formyl group / amino group is 0.1-500.
  • the molar ratio of formyl group / amino group is 1-400, 1_350 in another specific example, 1-300 in another specific example, and another specific example. In the example, 10–300.
  • the first component and the second component may be applied to the adherend (skin surface inside and outside the living body) simultaneously or sequentially for a medical effect (medical use).
  • a medical effect medical use
  • saline or distilled water may be sprayed in order to gel the first and second components.
  • the medical use is selected from the group consisting of adhesion, layer deposition, application, adhesion prevention, wound coating and hemostasis of biological tissue.
  • the composition of the present invention provides the first component and the second component in a form contained in the same container, or in a form separately contained in a separate container.
  • the present invention provides a biodegradable medical adhesive or sealant composition comprising an oxidized glycosaminoglycan and a polyamine.
  • composition of the present invention shows an improved effect in biodegradable coating properties, gelation time, hemostatic capacity, adhesion and water absorption.
  • composition of the present invention can be utilized for various medical applications in which medical adhesives or sealants can be used, such as adhesion, layering, coating, adhesion prevention, wound coating, leakage prevention, and hemostasis of biological tissues.
  • Figure 1 shows the results of the analysis of oxidized hyaluronic acid using an FT-IR spectrometer.
  • FIG. 3 is a photograph showing the results of gelation evaluation of the first and second component mixtures mixed in different weight ratios.
  • Figure 4 is a photograph showing a comparison of the gelation time of the adhesive and sealant composition of the present invention and the conventional adhesive composition (Lydex).
  • 5 is a graph showing the adhesive strength of the adhesive and sealant composition of the present invention and the conventional adhesive composition (Litex).
  • Figure 6 is a photograph showing the results (mucoadhesion and hemostatic capacity) after applying the adhesive and sealant composition of the present invention, or a conventional adhesive composition (Lydex) to the bleeding site after gastric mucosal resection.
  • Figure 7 is a photograph showing the gelation state and gelation time of the oxidized glycosaminoglycans and polyamine mixtures and the gelation according to pH.
  • FIGS. 8 to 11 show the results of comparative experiments between the adhesive and sealant composition of the present invention and the conventional hemostatic agent (Ar i sta TM AH) performed on the mesenchymal resection model, nephrectomy model, gastric mucosal resection model, and blood vessel application model.
  • . 12 is a graph quantitatively showing the results of FIGS. 8 to 11.
  • the oxidized hyaluronic acid obtained here was lyophilized for at least 4 days, then pulverized and passed through a 500 ⁇ sized mesh to obtain an oxidized hyaluronic acid having a diameter of about 500 im or less.
  • Oxidized hyaluronic acid was analyzed using an FT-IR spectrometer (Cary 640, Agilent Technologies, USA), and the substituent was confirmed at 4000-400 cm ' Hresolution 4 cm- 1 (FIG. 1).
  • chitosan, protamine, PEI, polylysine, spermine, spermidine, and albumin were used as second components, and pH adjusters (acid, acid, base, base salt, etc.) from 5% by weight or more of aqueous polyamine solution After adjusting the pH to 8.5, 9.0, 9.5 and 10 using the same as in the case of the oxidized hyaluronic acid / oxidized chondroitin sulfate, the powder obtained after freeze drying was used.
  • Example 1 The first and second components obtained in Example 1 were mixed in different weight ratios (1: 1, 2: 1,
  • LYDEX showed an average adhesive strength of 53.6 gf even though a small amount (500 water was applied) of LYDEX compared to the oxidized hyaluronic acid mixture. Measured at 65.9 and 67.5 gf (FIG. 5 and Table 6).
  • a mucosectomy-induced gastric bleeding model in rabbits was constructed as follows.
  • the rabbits were fasted for 24 hours prior to surgery and anesthetized by intramuscular injection of a combination of ketamine (4.2 mg / kg) and silazine (11.7 mg / kg).
  • the upper abdomen of the rabbit was incised to expose the stomach and incision about 5-7 cm along the great curvature of the stomach.
  • Isotonic saline 200 was injected into the submucosal layer of the stomach, and the swollen gastric mucosa was excised using surgical scissors. The diameter of the excised site was about 7-10 mm 3.
  • the degree of substitution (oxidation degree) of hyaluronic acid and chondroitin sulfate was confirmed by NaOH titration. Specifically, 17.5 g of hydroxylamine hydrochloride and 0.05% methyl orange 6 11 were mixed in 994 ⁇ distilled water to make a 0.25 1 ⁇ 1 / £ hydroxylamine hydrochloride solution and the pH was titrated to 4. 0.1 g of hyaluronic acid black silver chondroitin sulfate was dissolved in 25 m of the above solution and titrated again to pH 4 with 0.1 ⁇ / ⁇ sodium hydroxide. Substitution degree (%) was calculated using the following formula, and the results were as shown in Tables 8 and 9. Equation 3
  • chitosan, protamine PEI, polylysine, spermine, spermidine, and albumin were selected as the second component, and as a second component, to find out whether the gelation was performed according to pH.
  • aqueous solution after adjusting the pH to various ranges (5.5-6.4, 6.5-7.4, 7.5-8.4, 8.5-9.4, 9.5-10.4, 10.5-11), the same as in the case of the oxidized hyaluronic acid / oxidized chondroitin sulfate
  • the powder obtained after freeze drying was used. Oxidized hyaluronic acid / oxidized chondroitin sulfate of the first component and polyamine were mixed.
  • Anesthesia was injected by injection of a ketamine and lump mixture into the abdominal cavity of male SD rats weighing 200-300 g, and the central upper abdomen was incised approximately 3-4 cm in length or width.
  • the mesenchyme was exposed using a wet gauze into the incision of the incised abdomen, and the portal vein and hepatic artery were ligated with a vascular clip. About 1 cm away from the edge of the mesenchyme was excised with surgical scissors and 50-100 mg of UI-SAH was applied.
  • Ar i sta TM AH (Medafor Inc., USA) was applied. After application, the ligated clip was removed after ligating, and the amount of bleeding was measured using sterile gauze.
  • Anesthesia was injected by injection of a ketamine and lump mixture into the abdominal cavity of male SD rats weighing 200-300 g, and the central upper abdomen was incised about 5-6 cm vertically or horizontally.
  • the portal vein was exposed after moving other organs to the left axis through the incised abdominal gap.
  • Two sites above and below the portal vein were ligated with a vascular clip.
  • UI-SAH 50-100 mg was applied after puncturing the portal vein using an 18 gauge needle.
  • Ar i sta TM AH (Medafor Inc., USA) was applied. After application, the ligated clip was removed after removing the ligated clip, and the bleeding amount was measured using sterile gauze.

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Abstract

The present invention provides a biodegradable medical adhesive or a sealant composition containing an oxidized glycosaminoglycan and a polyamine. The composition of the present invention exhibits improved effects in biodegradation, coating property, gelation time, hemostatic capacity, adhesive force, moisture absorptive capacity and the like, and thus can be applied to various medical uses in which a medical adhesive or sealant can be used, such as biotissue adhesion, filling, coating, adhesion prevention, wound coating, leakage prevention and hemostasis.

Description

【명세서】  【Specification】
[발명의 명칭]  [Name of invention]
생분해성 의료용 접착제 또는 실란트 조성물 [기술 분야】  Biodegradable Medical Adhesive or Sealant Composition [Technical Field]
본 발명은 포밀기를 갖는 산화 글리코사미노글리칸과 폴리아민을 포함하는 생분해성 의료용 접착제 또는 실란트 조성물에 관한 것이다.  The present invention relates to a biodegradable medical adhesive or sealant composition comprising an oxidized glycosaminoglycan having a formyl group and a polyamine.
【배경 기술】 [Background technology]
생체 접착제 (bioadhesive) 및 실란트는 수술시 조직올 봉합 또는 도포하거나， 출혈 방지제 (지혈) , 체액과 혈액 차단제 등으로 사용되는 것으로， 피부에 접촉하므로 생체 적합성이 요구되며, 생체 내에서 독성과 위해성이 없어야 하고， 생분해성이야 하며, 생체의 치유를 방해하지 않아야 한다.  Bioadhesives and sealants are used to seal or apply tissues during surgery, or to be used as anti-bleeding agents (hemostasis), body fluids and blood blockers. Bioadhesives and sealants require biocompatibility as they come into contact with the skin. It must be absent, biodegradable, and not interfere with the healing of living organisms.
현재 실용화되고 있는 의료용 접착 소재로는 시아노아크릴레이트계, 피브린 글루계, 젤라틴 글루계, 폴리우레탄계 등이 있으며， 미국의 Closure Medi cal사는 Dermabond라는 옥틸시아노아크릴레이트의 의료용 조직접착제를 상품화하여 1997년 8월 유럽공동체의 판매승인을 얻은 데 이어 1998년 미국 FDA로부터 승인을 받은바 있다. 그러나， 시아노아크릴레이트계 접착제는 고화물이 유연성이 부족하고 단단하므로 창상 치유를 방해하는 경우가 있고 또한 생체 내에서 분해되기 어려우므로 피포화 (被包化)되어 이물질이 되기 쉬운 등의 문제가 있었다. 또한, 피브린 글루는 접착력이 상당히 낮으므로， 생성된 피브린 명어리가 조직으로부터 떨어지는 경우가 있으며, 혈액 제제이므로 바이러스 감염이 우려되는 등의 문제가 있었다.  Medical adhesive materials currently in use include cyanoacrylates, fibrin glues, gelatin glues, and polyurethanes. Closure Medi Cal of the United States commercialized a medical tissue adhesive of octylcyanoacrylate called Dermabond in 1997. It was approved by the US FDA in 1998 after it was approved by the European Community in August. However, cyanoacrylate-based adhesives may interfere with wound healing because the solids lack rigidity and rigidity, and are difficult to decompose in vivo, and thus are easily encapsulated and become foreign matters. there was. In addition, since fibrin glue has a considerably low adhesive force, the generated fibrin fish may drop from the tissue, and since the blood product is a blood product, there is a problem of a viral infection.
상기 의료용 접착제 외에， 한국공개특허 게 10-2009-0083484호에는 동결건조 후 기계적으로 분쇄하여 제조된 알데히드화 텍스트란 분말과 ε - 폴리— L-리신 분말을 포함하는 의료용 2 성분형 접착제 (상품명: LYDEX , 라이덱스)가 개시되어 있다. 상기 2 성분형 접착제는 분말상의 의료용 접착제라는 점에 특징이 있으나， 겔 분해 시간이 비교적 오래 소요되고， 수분 흡수력에서 만족할만한 효과를 얻지 못하는 문제가 있다. 따라서， 분해 시간， 접착력 및 수분 흡수력 등에 있어 개선된 특징을 갖는 새로운 의료용 접착제에 대한 요구가 높아지고 있다. 본 명세서 전체에 걸쳐 다수의 는문 및 특허문헌이 참조되고 그 인용이 표시되어 있다. 인용된 논문 및 특허문헌의 개시 내용은 그 전체로서 본 명세서에 참조로 삽입되어 본 발명이 속하는 기술 분야의 수준 및 본 발명의 내용이 보다 명확하게 설명된다. In addition to the medical adhesive, Korean Patent Publication No. 10-2009-0083484 discloses a medical two-component adhesive comprising an aldehyde-ized textan powder and an ε-poly—L-lysine powder prepared by mechanical grinding after lyophilization. LYDEX, Rydex) is disclosed. The two-component adhesive is characterized in that it is a powdered medical adhesive, it takes a relatively long time to decompose the gel, there is a problem that does not obtain a satisfactory effect in the water absorption. Thus, new products with improved characteristics in terms of decomposition time, adhesion and water absorption There is an increasing demand for medical adhesives. Throughout this specification, numerous citations and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.
【발명의 내용】 [Content of invention]
【해결하려는 과제】  [Problem to solve]
본 발명자들은 체액 및 혈액이 존재하는 체내 부위에서도 충분한 접착， 피복 및 지혈을 수행할 수 있고, 특히 다른 생분해성 고분자에 비해 많은 양의 수분을 흡수할 수 있으며, 체내에서 스스로 분해 가능한 생분해성 의료용 접착제 및 실란트를 개발하기 위하여 연구 노력하였다. 그 결과, 포밀기가 도입되어 산화된 글리코사미노글리칸과 폴리아민을 함께 사용함으로써 생체 조직의 접착， 층전, 도포， 유착 방지， 창상 피복 및 지혈 등을 효과적으로 할 수 있음을 확인함으로써, 본 발명을 완성하게 되었다.  The present inventors can perform sufficient adhesion, coating and hemostasis even in the part of the body where body fluid and blood are present, and can absorb a large amount of water, especially compared to other biodegradable polymers, and can decompose itself in the body. And research efforts were made to develop sealants. As a result, the present invention is confirmed that the use of a formyl group and oxidized glycosaminoglycan and polyamine together can effectively bond, layer, apply, prevent adhesion, wound coating and hemostasis of living tissues. It was completed.
따라서， 본 발명의 목적은 의료용 접착제 또는 실란트 (또는 의료용 접착제 또는 실란트 조성물)를 제공하는 데 있다.  Accordingly, an object of the present invention is to provide a medical adhesive or sealant (or a medical adhesive or sealant composition).
본 발명의 다른 목적은 생체 조직의 접착， 층전, 도포， 유착 방지, 창상 피복 또는지혈방법을 제공하는 데 있다. 본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명， 청구범위 및 도면에 의해 보다 명확하게 된다.  Another object of the present invention is to provide a method for adhesion, layer deposition, application, adhesion prevention, wound coating or hemostasis of living tissue. Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.
【과제의 해결 수단】 [Measures of problem]
본 발명의 일 양태에 따르면， 본 발명은 다음을 포함하는 생분해성 의료용 접착제 또는 실란트 (또는 의료용 접착제 또는 실란트 조성물)를 제공한다:  According to one aspect of the invention, the invention provides a biodegradable medical adhesive or sealant (or medical adhesive or sealant composition) comprising:
(a) 포밀기가 도입되어 산화된 산화 글리코사미노글리칸을 포함하는 제 1 성분; 및 (b) 2 이상의 아미노기를 갖는 폴리아민을 포함하는 제 2 성분， 상기 제 2 성분의 수용액 상태에서의 pH는 8.5 내지 11.0임. (a) a first component comprising an oxidized glycosaminoglycan in which a formyl group is introduced and oxidized; And (b) A second component comprising a polyamine having two or more amino groups, wherein the pH in the aqueous solution state of the second component is 8.5 to 11.0.
본 발명의 다른 일 양태에 따르면, 본 발명은 다음을 포함하는 생분해성 의료용 접착제 또는 실란트 (또는 의료용 접착제 또는 실란트 조성물)를 제공한다:  According to another aspect of the invention, the invention provides a biodegradable medical adhesive or sealant (or medical adhesive or sealant composition) comprising:
(a) 포밀기가 도입되어 산화된 산화 글리코사미노글리칸을 포함하는 제 1 성분; 및  (a) a first component comprising an oxidized glycosaminoglycan in which a formyl group is introduced and oxidized; And
(b) 2 이상의 아미노기를 갖는 폴리아민을 포함하는 제 2 성분， 상기 제 2 성분의 수용액 상태에서의 pH는 8.5 내지 11.0이고,  (b) a second component comprising a polyamine having two or more amino groups, the pH in the aqueous solution state of the second component is 8.5 to 11.0,
상기 제 1 및 제 2 성분을 흔합한 경우 포밀기 /아미노기의 몰비는 When the first and second components are mixed, the molar ratio of formyl group / amino group is
0. 1 내지 500임 . 0.1 to 500.
본 발명의 또 다른 일 양태에 따르면， 본 발명은 상기 생분해성 의료용 접착제 또는 실란트 (또는 의료용 접착제 또는 실란트 조성물)를 접착， 층전, 도포， 유착 방지, 창상 피복 또는 지혈이 필요한 생체 조직에 적용하는 단계를 포함하는 생체 조직의 접착, 층전， 도포， 유착 방지， 창상 피복 또는 지혈방법을 제공한다. 본 발명자들은 체액 및 혈액이 존재하는 체내 부위에서도 충분한 접착， 피복및 지혈을 수행할 수 있고， 특히 다른 생분해성 고분자에 비해 많은 양의 수분올 흡수할 수 있으며， 체내에서 스스로 분해 가능한 생분해성 의료용 접착제 및 실란트를 개발하기 위하여 연구 노력하였다. 그 결과， 포밀기가 도압되어 산화된 글리코사미노글리칸과 폴리아민을 함께 사용함으로써 생체 조직의 접착， 층전， 도포， 유착 방지, 창상 피복 및 지혈 둥을 효과적으로 할 수 있음을 확인하였다.  According to another aspect of the invention, the present invention is the step of applying the biodegradable medical adhesive or sealant (or medical adhesive or sealant composition) to a biological tissue that requires adhesion, layering, application, adhesion prevention, wound coating or hemostasis It provides a method of adhesion, layer deposition, coating, adhesion prevention, wound coating or hemostasis of a biological tissue comprising a. The present inventors can perform sufficient adhesion, coating and hemostasis even in the part of the body where body fluids and blood are present, and in particular, can absorb a large amount of water compared to other biodegradable polymers and can decompose itself in the body. And research efforts were made to develop sealants. As a result, it was confirmed that by using a polyamine with glycosaminoglycan oxidized by the formyl group, the adhesion, layer formation, coating, adhesion prevention, wound coating and hemostasis of biological tissues can be effectively performed.
본 명세서에서 의료용 접착제 또는 실란트와 이의 조성물은 상호 교환적으로 사용된다.  Medical adhesives or sealants and compositions thereof are used interchangeably herein.
하기 실시예에서 확인한 바와 같이 , 본 발명의 조성물은 기존의 의료용 2 성분형 접착제 (LYDEX) 보다 겔 형성 시간， 점착력 및 수분 흡수력에서 개선된 효과를 나타내었으며 (표 5 내지 7 참조)， 지혈 효과에 있어서도 기존의 지혈제 (Ar i staTMAH) 보다 더 우수한 효과를 나타내었다 (도 12 참조) . 이러한 결과들은 제 1 성분과 제 2 성분의 조합인 본 발명의 조성물이 생체 조직의 접착， 충전， 도포, 유착 방지， 창상 피복 및 지혈 등의 의료 용도에 있어 우수한 물성을 나타내어， 본 발명의 조성물을 이들 의약 용도로 사용할 수 있음올 보여준다 . As confirmed in the following examples, the composition of the present invention showed an improved effect on gel formation time, adhesion and water absorption than the conventional medical two-component adhesive (LYDEX) (see Tables 5 to 7), In addition, it showed a better effect than the conventional hemostatic agent (Ar i sta TM AH) (see Fig. 12). These results indicate that the combination of the first component and the second component The composition exhibits excellent physical properties in medical applications such as adhesion, filling, application, adhesion prevention, wound coating and hemostasis of biological tissues, showing that the composition of the present invention can be used in these pharmaceutical applications.
본 발명의 조성물은 산화 글리코사미노글리칸을 제 1 성분으로 포함한다. 상기 용어, "산화 글리코사미노글리칸 "은 글리코사미노글리칸에 포밀기 (-CH0)가 도입되어 산화되었음을 의미한다. 상기 글리코사미노 글리칸은 핵소사민을 포함한 2당의 반복구조를 갖는 다당이라는 점에서 단당의 0-글리코시드 결합인 글루칸과는 구별된다.  The composition of the present invention comprises oxidized glycosaminoglycan as the first component. The term "oxidized glycosaminoglycan" means that a formyl group (-CH0) is introduced and oxidized to glycosaminoglycan. The glycosamino glycan is distinguished from glucan, which is a 0-glycoside bond of a monosaccharide, in that it is a polysaccharide having a repeating structure of a disaccharide including nucleosamine.
이러한 포밀기의 도입은 과요오드산 산화법으로 실시할 수 있다. 예를 들어, 글리코사미노글리칸올 과요오드산 또는 과요오드산염으로 산화하여 무수 포도당 단위 (당 잔기 ) 당 적정 개수 (예를 들어， 0.01 내지 0.95개)의 포밀기가 도입된 산화 글리코사미노글리칸을 얻을 수 있다.  Introduction of such a formyl group can be performed by a periodic acid oxidation method. For example, oxidized glycosaminoglyco introduced with the appropriate number (eg, 0.01 to 0.95) formyl groups per anhydrous glucose unit (sugar moiety) by oxidation with glycosaminoglycanol periodic acid or periodicate You can get a space.
본 글리코사미노글리칸의 산화도는
Figure imgf000006_0001
에 따라 계산되며, 10 내지 99.5%의 값을 갖는다. 이와 같은 산화도를 갖는 산화 글리코사미노글리칸은 제 2 성분과 조합사용 시, 체내의 혈액과 체액을 신속하게 흡수하여 빠른 시간 안에 겔화할 수 있다. Oxidation degree of this glycosaminoglycan
Figure imgf000006_0001
Calculated according to, having a value from 10 to 99.5%. Oxidized glycosaminoglycans having such a degree of oxidation, when used in combination with the second component, can quickly absorb the blood and body fluids in the body and gelate quickly.
하나의 특정예에 따르면， 상기 산화 글리코사미노글리칸의 산화도는 According to one specific example, the oxidation degree of the oxidized glycosaminoglycan is
10 내지 60%이고, 다른 특정예에서는 10 내지 55%이며， 또 다른 특정예에서는 10 내지 50<¾이고， 또 다른 특정예에서는 10 내지 45%이며, 또 다른 특정예에서는 10 내지 40%이다. 일예로, 상기 게 1 성분에 1 종류 이상의 산화 글리코사미노글리칸이 포함되며， 이 산화 글리코사미노글리칸이 산화 히알루론산인 경우, 상기 산화 히알루론산의 산화도는 10-20%일 수 있다. 10 to 60%, 10 to 55% in another specific example, 10 to 50 <¾ in another specific example, 10 to 45% in another specific example, and 10 to 40% in another specific example. For example, when one or more types of oxidized glycosaminoglycans are included in the crab component, and the oxidized glycosaminoglycans are oxidized hyaluronic acid, the oxidation degree of the oxidized hyaluronic acid may be 10-20%. .
상기 글리코사미노글리칸의 산화도 측정은 NaOH 점정법으로 실시할 수 있다. 예를 들어, 17.5 g의 하이드록실아민 하이드로클로라이드와 0.05% 메틸 오렌지 6 을 994 의 증류수에 흔합하여 0 .25 ιηοΐ/ ί의 하이드록실아민 하이드로클로라이드 용액을 만들고 ρΗ를 4로 적정한 다음， 산화 글리코사미노글리칸 0. 1 g을 25 ^의 상기 용액에 녹이고， 0. 1 \/ i 수산화나트륨을 이용하여 다시 pH 4로 적정한 후， ， ， 一 ， ， ，
Figure imgf000007_0001
Oxidation degree measurement of the glycosaminoglycan can be carried out by NaOH scoring method. For example, 17.5 g of hydroxylamine hydrochloride and 0.05% methyl orange 6 are mixed in 994 distilled water to form a solution of 0.25 ιηοΐ / ί hydroxylamine hydrochloride, titrated ρΗ to 4, and then Dissolve 0.1 g of minoglycan in 25 ^ of the above solution, titrate it again to pH 4 with 0.1 \ / i sodium hydroxide, ， ， 一 ， ， ，
Figure imgf000007_0001
도 (%)를 계산한다. Calculate the degree (%).
본 발명의 일구현예에 따르면, 상기 산화 글리코사미노글리칸은 무수 포도당 단위 (당 잔기) 당 0.01 내지 0.95개의 포밀기를 갖는다.  According to one embodiment of the invention, the oxidized glycosaminoglycan has 0.01 to 0.95 formyl groups per anhydrous glucose unit (sugar residues).
본 발명의 일구현예에 따르면， 상기 산화 글리코사미노글리칸은 산화 히알루론산, 산화 콘드로이틴 설페이트， 산화 콘드로이린, 산화 더마탄 설페이트， 산화 헤파란 설페이트， 산화 헤파린 및 산화 케라탄 설페이트로 구성된 군으로부터 선택된다.  According to one embodiment of the present invention, the oxidized glycosaminoglycan is composed of oxidized hyaluronic acid, oxidized chondroitin sulfate, oxidized chondroitin, dermatan sulphate, oxidized heparan sulphate, heparin oxidized and keratan oxidized. Is selected from.
본 발명에 따르면， 특정 분자량의 글리코사미노글리칸을 사용함으로써 접착제 /실란트 조성물의 겔화 형성능, 겔화 상태 유지시간, 겔의 탄성 등을 적절히 조절할 수 있다.  According to the present invention, by using a glycosaminoglycan having a specific molecular weight, the gelling ability of the adhesive / sealant composition, the gelation state retention time, the elasticity of the gel, and the like can be appropriately controlled.
본 발명의 일구현예에 따르면， 상기 산화 글리코사미노글리칸을 얻는데 사용되는 글리코사미노글리칸은 1 , 000 내지 500만의 분자량올 갖는다. 일예로， 상기 산화 글리코사미노글리칸을 얻는데 사용되는 글리코사미노글리칸은 1만 내지 400만, 5만 내지 350， 10만 내지 350， 10만 내지 300만， 10만 내지 250만, 10만 내지 200만， 또는 10만 내지 160만의 분자량을 가질 수 있다.  According to one embodiment of the present invention, the glycosaminoglycan used to obtain the oxidized glycosaminoglycan has a molecular weight of 1,000 to 5 million. For example, the glycosaminoglycan used to obtain the oxidized glycosaminoglycan is 10,000 to 4 million, 50,000 to 350, 100,000 to 350, 100,000 to 3 million, 100,000 to 2.5 million, 100,000 To 2 million, or 100,000 to 1.6 million molecular weight.
본 발명의 일구현예에 따르면, 상기 제 1 성분에는 10만 내지 According to an embodiment of the present invention, the first component is 100,000 to
200만의 분자량을 갖는 산화 히알루론산이 포함된다. 하나의 특정예에 따르면， 상기 산화 히알루론산의 분자량은 10만 내지 160만이다. Oxidized hyaluronic acid with a molecular weight of 2 million is included. According to one specific example, the molecular weight of the oxidized hyaluronic acid is 100,000 to 1.6 million.
본 발명의 일구현예에 따르면, 상기 제 1 성분에는 2 종류 이상의 산화 글리코사미노글리칸이 포함된다.  According to an embodiment of the present invention, the first component includes two or more kinds of oxidized glycosaminoglycans.
본 발명의 일구현예에 따르면， 이들 산화 글리코사미노글리칸의 중량비는 1 : 0. 5-5이다. 하나의 특정예에 따르면， 상기 산화 글리코사미노글리칸의 중량비는 1 : 0.5-4이고， 다른 특정예에서는 1 : 0. 5- According to one embodiment of the present invention, the weight ratio of these oxidized glycosaminoglycans is 1: 0.5-5. According to one specific example, the weight ratio of the oxidized glycosaminoglycan is 1: 0.5-4, and in another specific example 1: 0.5.
3.5이며， 또 다른 특정예에서는 1 : 0. 5-3이고， 또 다른 특정예에서는 1 : 0.5-3.5, and in another specific example, 1: 0.5-3, and in another specific example, 1: 0.5-
2.5이고, 또 다른 특정예에서는 1 : 0. 5-2이며, 또 다른 특정예에서는 1 : 0.5-2.5, and in another specific example, 1: 0.5-2, and in another specific example, 1: 0.5-
1 .5이다. 1.5.
본 발명의 일구현예에 따르면， 상기 2 종류 이상의 산화 글리코사미노글리칸은 산화 히알루론산 및 산화 콘드로이틴 설페이트이다. 상기 산화 히알루론산과 산화 콘드로이틴 설페이트의 중량비는 1:0.5-5， 1:0.5-4， 1:0.5-3.5， 1:0.5-3， 1:0.5-2.5， 1:0.5-2， 1:0.5-1.5 또는 1:0.8-According to one embodiment of the present invention, the two or more kinds of oxidation Glycosaminoglycans are oxidized hyaluronic acid and oxidized chondroitin sulfate. The weight ratio of the hyaluronic acid oxide and chondroitin oxide is 1: 0.5-5, 1: 0.5-4, 1: 0.5-3.5, 1: 0.5-3, 1: 0.5-2.5, 1: 0.5-2, 1: 0.5 -1.5 or 1: 0.8-
I.2일 수 있다. May be I.2.
본 발명의 일구현예에 따르면, 상기 산화 히알루론산의 산화도는 10- 이다. 하나의 특정예에 따르면， 상기 산화 히알루론산의 산화도는 12- 40%이고， 다른 특정예에서는 12-38¾>이며， 또 다른 특정예에서는 13-3 이다. 본 발명의 일구현예에 따르면， 상기 산화 콘드로이틴 설페이트의 산화도는 10-55%이다. 하나의 특정예에 따르면， 상기 산화 콘드로이린 설페이트의 산화도는 10-50%이고， 다른 특정예에서는 10-45%이며， 또 다른 특정예에서는 10-40%이고, 또 다른 특정예에서는 10-35%이다.  According to an embodiment of the present invention, the oxidation degree of the oxidized hyaluronic acid is 10-. According to one specific example, the oxidation degree of the oxidized hyaluronic acid is 12-40%, in another specific example 12-38¾>, and in another specific example 13-3. According to one embodiment of the present invention, the oxidation degree of the oxidized chondroitin sulfate is 10-55%. According to one specific example, the oxidation degree of the chondroitin sulfate is 10-50%, in another specific example 10-45%, in another specific example 10-40%, in another specific example 10 -35%.
본 발명의 일구현예에 따르면， 상기 제 1 성분은 분말상， 액상 또는 고체상 (예컨대， 펠렛 형태)이다. 일예로， 분말상의 제 1 성분은 산화 글리코사미노글리칸 -함유 용액을 건조 (예컨대， 분무건조, 동결건조 등)한 후 분쇄 (예컨대， 기계적 분쇄)하여 수득할 수 있다.  According to one embodiment of the invention, the first component is in powder, liquid or solid (eg pellet form). For example, the powdery first component may be obtained by drying (eg, spray drying, lyophilization, etc.) of an oxidized glycosaminoglycan-containing solution followed by grinding (eg, mechanical grinding).
본 발명의 조성물은 활성성분으로서 제 1 성분 외에 2 이상의 아미노기를 갖는 폴리아민을 포함하는 제 2 성분을 더 포함한다. 상기 제 2 성분은 수용액 상태에서 8.5-11.0의 pH를 나타낸다. 하기 실시예에서 확인한 바와 같이， 상기 제 2 성분의 수용액 상태에서의 pH가 8.5-11.0인 경우에 수초 내로 겔화가 이루어질 수 있다 (도 7 참조).  The composition of the present invention further comprises a second component comprising a polyamine having two or more amino groups in addition to the first component as the active component. The second component exhibits a pH of 8.5-11.0 in aqueous solution. As confirmed in the following examples, when the pH in the aqueous solution state of the second component is 8.5-11.0, gelation may be performed within a few seconds (see FIG. 7).
하나의 특정예에 따르면， 상기 제 2 성분은 수용액 상태에서 9.0- According to one specific example, the second component is 9.0- in an aqueous solution state.
II.0의 pH를 나타낸다. PH of II.0 is shown.
본 발명의 일구현예에 따르면, 상기 플리아민은 2차 및 /또는 3차 아미노기를 더 가질 수 있다.  According to one embodiment of the invention, the pleamine may further have a secondary and / or tertiary amino group.
본 발명의 일구현예에 따르면， 상기 폴리아민을 포함하는 제 2 성분은 분말상, 액상 또는 고체상 (예컨대, 펠렛)이다. 일예로, 분말상의 제 2 성분은 폴리아민 -포함 용액을 건조한 후 분쇄하여 수득할 수 있다. 이때， 상기 폴리아민 -포함 용액은 제 2 성분의 수용액 상태에서의 pH 범위가 8.5- 11.0이 되도록 pH 조절제를 더 포함할 수 있다. 상기 pH 조절제로의 예로는， 아세트산， 시트르산, 숙신산， 글루타르산， 말산, 푸마르산 및 말레산과 같은 1가 또는 다가의 카본산 화합물 또는 그 무수물 등을 들 수 있다. 본 발명의 일구현예에 따르면， 상기 폴리아민은 폴리리신， 키토산， 알부민 , 푸트레신 (putrescine) , 카다베린 (cadaver ine)， 스페르미딘 (spermidine) , 스페르민 (spermine) , .프로타민 및 PEI (Polyethylenimine)로 구성된 군으로부터 선택된다. According to one embodiment of the invention, the second component comprising the polyamine is in powder, liquid or solid (eg, pellets). In one example, the powdery second component can be obtained by drying and grinding the polyamine-comprising solution. At this time, the polyamine-containing solution may further include a pH adjusting agent so that the pH range in the aqueous solution state of the second component is 8.5-11.0. Examples of the pH regulator include monovalent or polyvalent carboxylic acid compounds such as acetic acid, citric acid, succinic acid, glutaric acid, malic acid, fumaric acid and maleic acid, or anhydrides thereof. According to one embodiment of the invention, the polyamine is polylysine, chitosan, albumin, putrescine (putrescine), cadaverine (cadaver ine), spermidine, spermine (spermine), protamine and PEI (Polyethylenimine) is selected from the group consisting of.
본 발명의 일구현예에 따르면, 상기 폴리아민은 100 이상의 분자량을 갖는다. 일예로， 상기 폴리아민의 분자량은 1 ,000 내지 20만일 수 있다. 본 발명의 일구현예에 따르면， 상기 폴리아민은 폴리 -L-리신이다. 상기 폴리 -L-리신은 미생물 (예컨대， 스트랩토마이세스 아불러스) 또는 효소를 사용하여 생산된 ε -폴리 -L-리신일 수 있다.  According to one embodiment of the invention, the polyamine has a molecular weight of 100 or more. For example, the molecular weight of the polyamine may be 1,000 to 200,000. According to one embodiment of the invention, the polyamine is poly-L-lysine. The poly-L-lysine may be ε-poly-L-lysine produced using microorganisms (eg, Straptomyces abulus) or enzymes.
본 발명의 일구현예에 따르면， 상기 제 2 성분은 폴리아민 외에 ρΗ 조절제를 더 포함할 수 있다.  According to one embodiment of the invention, the second component may further comprise a ρΗ regulator in addition to the polyamine.
본 발명의 일구현예에 따르면, 본 발명의 조성물은 약물을 더 포함할 수 있다. 일예로 상기 약물은 제 2 성분에 포함될 수 있다. 상기 약물은 According to one embodiment of the present invention, the composition of the present invention may further comprise a drug. In one example, the drug may be included in the second component. The drug is
1개 이상의 아민기를 가질 수 있으며, 이러한 약물의 예로는， 안트 라사이클린계열 약물, 젬시타빈， 반코마이신， 폴리미신， 메토트렉세이트， 단백질 약물 및 펩타이드 약물 등을 들 수 있다. 이 경우， 제 1 성분과 제It may have one or more amine groups, and examples of such drugs include anthracycline-based drugs, gemcitabine, vancomycin, polymycin, methotrexate, protein drugs and peptide drugs. In this case, the first component and the first
2 성분의 겔화 시 상기 약물의 아민기 역시 제 1 성분의 포밀기와 작용하여 세 성분이 함께 겔을 형성할 수 있고ᅳ 형성된 겔의 붕괴에 따라 약물이 서서히 방출되어 약리활성 활성을 나타낼 수 있다. In the gelling of the two components, the amine group of the drug may also act as the formyl group of the first component, and the three components may form a gel together.
본 발명의 일구현예에 따르면， 본 발명의 조성물은 여러 형태로 제형화 될 수 있고, 예를 들어， 분말상, 액상 또는 고체상 (예컨대， 펠렛 형태)인 제 1 성분과, 분말상， 액상 또는 고체상인 제 2 성분의 조합을 포함할 수 있다.  According to one embodiment of the present invention, the composition of the present invention may be formulated in various forms, for example, a first component which is in powder form, liquid form or solid form (for example, pellet form), and is in powder form, liquid form or solid form. Combinations of second components.
본 발명의 일구현예에 따르면， 본 발명의 조성물은 제 1 성분과 제 2 성분을 0.5-10 : 1의 중량비로 포함한다.  According to one embodiment of the present invention, the composition of the present invention comprises the first component and the second component in a weight ratio of 0.5-10: 1.
하나의 특정예에 따르면, 상기 제 1 성분과 계 2 성분은 0.5-8 : 1의 중량비로 포함되며, 다른 특정예에서는 0.5-6 : 1의 중량비， 또 다른 특정예에서는 0.5-4 : 1의 중량비， 또 다른 특정예에서는 0.5-3 : 1의 중량비， 또 다른 특정예에서는 0.5-2: 1의 중량비， 또 다른 특정예에서는 0.5- 1.5: 1의 중량비， 또 다른 특정예에서는 0.8-1.5: 1의 중량비， 또 다른 특정예에서는 0.8-1.2 : 1의 중량비로 포함된다. 본 발명에 따르면， 상기 제 1 성분의 포밀기와 제 2 성분의 아미노기의 비율을 조절함으로써 겔 형성시간， 생성 겔의 분해시간 등을 조절할 수 있다. According to one specific example, the first component and the second component are included in a weight ratio of 0.5-8: 1, in another specific example, a weight ratio of 0.5-6: 1, and in another specific example, 0.5-4: 1 Weight ratio, in another specific example a weight ratio of 0.5-3: 1, in another specific example, a weight ratio of 0.5-2: 1, in another specific example, a weight ratio of 0.5-1.5: 1, and in another specific example, 0.8-1.5: Weight ratio of 1, and in another specific example, weight ratio of 0.8-1.2: 1. According to the present invention, by controlling the ratio of the formyl group of the first component and the amino group of the second component it is possible to control the gel formation time, the decomposition time of the resulting gel and the like.
본 발명의 일구현예에 따르면， 상기 제 1 및 제 2 성분을 흔합한 경우 포밀기 /아미노기의 몰비는 0. 1-500이다.  According to one embodiment of the present invention, when the first and second components are mixed, the molar ratio of formyl group / amino group is 0.1-500.
하나의 특정예에 따르면， 상기 제 1 및 제 2 성분을 흔합한 경우 포밀기 /아미노기의 몰비는 1-400이고， 다른 특정예에서는 1_350이며， 또 다른 특정예에서는 1-300이고， 또 다른 특정예에서는 10— 300이다.  According to one specific example, when the first and second components are mixed, the molar ratio of formyl group / amino group is 1-400, 1_350 in another specific example, 1-300 in another specific example, and another specific example. In the example, 10–300.
상기 제 1 성분과 제 2 성분은 의료 효과 (의료 용도)를 위하여, 동시에 혹은 순차적으로 피착체 (생체 내외의 피부면)에 도포될 수 있다. 이때ᅳ 경우에 따라 상기 제 1 및 제 2 성분의 겔화를 위하여 식염수 혹은 증류수를 분사할 수 있다.  The first component and the second component may be applied to the adherend (skin surface inside and outside the living body) simultaneously or sequentially for a medical effect (medical use). In this case, saline or distilled water may be sprayed in order to gel the first and second components.
본 발명의 일구현예에 따르면, 상기 의료 용도는 생체 조직의 접착, 층전， 도포, 유착 방지, 창상 피복 및 지혈로 구성된 군으로부터 선택된다. 본 발명의 조성물은 제 1 성분과 제 2 성분을 동일한 용기에 담겨진 형태로 제공하거나， 또는 별도의 용기에 분리되어 담겨진 형태로 제공한다.  According to one embodiment of the invention, the medical use is selected from the group consisting of adhesion, layer deposition, application, adhesion prevention, wound coating and hemostasis of biological tissue. The composition of the present invention provides the first component and the second component in a form contained in the same container, or in a form separately contained in a separate container.
【발명의 효과】 【Effects of the Invention】
본 발명의 특징ᅵ 및 이점을 요약하면 다음과 같다: It A summary of the features and advantages of the present invention is as follows:
( i ) 본 발명은 산화 글리코사미노글리칸과 폴리아민을 포함하는 생분해성 의료용 접착제 또는 실란트 조성물을 제공한다.  (i) The present invention provides a biodegradable medical adhesive or sealant composition comprising an oxidized glycosaminoglycan and a polyamine.
( i i ) 본 발명의 조성물은 생분해성 도포성， 겔화 시간， 지혈능， 접착력 및 수분 흡수력 등에서 개선된 효과를 나타낸다.  (i i) The composition of the present invention shows an improved effect in biodegradable coating properties, gelation time, hemostatic capacity, adhesion and water absorption.
( iii ) 본 발명의 조성물은 생체 조직의 접착, 층전， 도포, 유착 방지， 창상 피복， 누출방지 및 지혈 등 의료용 접착제 또는 실란트가 사용 가능한 다양한 의료 용도로 활용될 수 있다.  (iii) The composition of the present invention can be utilized for various medical applications in which medical adhesives or sealants can be used, such as adhesion, layering, coating, adhesion prevention, wound coating, leakage prevention, and hemostasis of biological tissues.
【도면의 간단한 설명】 [Brief Description of Drawings]
도 1은 FT-IR 분광계를 이용하여 산화 히알루론산을 분석한 결과를 보여준다.  Figure 1 shows the results of the analysis of oxidized hyaluronic acid using an FT-IR spectrometer.
도 2는 아미노기 갖는 제 2 성분을 이용한 흔합물의 겔화 모습과 겔화 시간을 보여주는 사진이다. 2 is a gelation state of the mixture using a second component having an amino group Photo shows gelation time.
도 3은 서로 다른 중량비로 흔합한 제 1 성분과 제 2 성분 흔합물의 겔화 평가 결과를 보여주는 사진이다.  3 is a photograph showing the results of gelation evaluation of the first and second component mixtures mixed in different weight ratios.
도 4는 본 발명의 접착제 및 실란트 조성물과 기존의 접착제 조성물 (라이덱스)의 겔화 시간을 비교하여 보여주는 사진이다.  Figure 4 is a photograph showing a comparison of the gelation time of the adhesive and sealant composition of the present invention and the conventional adhesive composition (Lydex).
도 5는 본 발명의 접착제 및 실란트 조성물과 기존의 접착제 조성물 (라이텍스)의 점착력을 비교하여 보여주는 그래프이다.  5 is a graph showing the adhesive strength of the adhesive and sealant composition of the present invention and the conventional adhesive composition (Litex).
도 6은 위점막 절제 후 출혈 부위에 본 발명의 접착제 및 실란트 조성물， 또는 기존의 접착제 조성물 (라이덱스)을 도포한 후의 결과 (점막 부착능력과 지혈능력)를 보여주는 사진이다.  Figure 6 is a photograph showing the results (mucoadhesion and hemostatic capacity) after applying the adhesive and sealant composition of the present invention, or a conventional adhesive composition (Lydex) to the bleeding site after gastric mucosal resection.
도 7은 산화 글리코스아미노글리칸과 폴리아민 흔합물의 겔화 모습과 겔화 시간， 그리고 pH에 따른 겔화 여부를 보여주는 사진이다.  Figure 7 is a photograph showing the gelation state and gelation time of the oxidized glycosaminoglycans and polyamine mixtures and the gelation according to pH.
도 8 내지 11은 간엽 절제술 모델, 신장 절제술 모델， 위점막 절제술 모델 및 혈관 출원 모델을 대상으로 실시한 본 발명의 접착제 및 실란트 조성물과 기존의 지혈제 (Ar i staTMAH) 사이의 비교실험 결과를 보여준다. 도 12는 도 8 내지 11의 결과를 정량적으로 보여주는 그래프이다. 8 to 11 show the results of comparative experiments between the adhesive and sealant composition of the present invention and the conventional hemostatic agent (Ar i sta TM AH) performed on the mesenchymal resection model, nephrectomy model, gastric mucosal resection model, and blood vessel application model. . 12 is a graph quantitatively showing the results of FIGS. 8 to 11.
【발명을 실시하기 위한 구체적인 내용】 [Specific contents to carry out invention]
이하， 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로서 , 본 발명의 요지에 따리- 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다. 실시예  Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, and according to the gist of the present invention-it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples. will be. Example
실시예 1. 의료용 접착제의 제조 ①  Example 1 Preparation of Medical Adhesive
(1) 산화 히알루론산 (CH0—HA; 제 1 성분)의 제조  (1) Preparation of Oxidized Hyaluronic Acid (CH0—HA; First Component)
분자량 7 kDa , 150 kDa , 1400 kDa 흑은 3000 kDa 의 히알루론산 (HA) 1 g 또는 3 g 을 소듐 페리오데이트 (NaI04 )가 녹아있는 150 m£의 물에 녹였다. 이때 소듐 페리오데이트의 농도와 반응조건을 하기 표 1 내지 4 와 같이 다르게 함으로써 산화도 (치환도 (DS) , 를 다르게 하였다. 반웅 플라스크를 15-70 °C에서 3-48 시간 동안 반웅시켰다. 반옹물을 증류수로 분획분자량 1-100 kDa 의 투석막을 이용하여 24 시간 동안 투석하였다. 여기에서 수득한 산화 히알루론산을 4 일 이상 동결건조 한 후， 분쇄하여 500 μια 크기의 메쉬를 통과시킴으로써 직경 약 500 im 이하의 산화 히알루론산을 수득하였다. Molecular weight 7 kDa, 150 kDa, 1400 kDa Black silver 1 g or 3 g of 3000 kDa hyaluronic acid (HA) was dissolved in 150 m £ of water in which sodium periodate (NaI 4 ) was dissolved. At this time, the concentration of sodium periodate and the reaction conditions were changed as shown in the following Tables 1 to 4 to change the degree of oxidation (substitution degree (DS), etc .. The reaction flask was reacted at 15-70 ° C. for 3-48 hours. Distilled water Dialysis was performed for 24 hours using a dialysis membrane having a fraction molecular weight of 1-100 kDa. The oxidized hyaluronic acid obtained here was lyophilized for at least 4 days, then pulverized and passed through a 500 μια sized mesh to obtain an oxidized hyaluronic acid having a diameter of about 500 im or less.
산화히알루론산  Hyaluronic Acid Oxide
'職 '職
Figure imgf000012_0001
산화 히알루론산을 FT-IR 분광계 (Cary 640, Agilent Technologies, USA)를 이용하여 분석한 결과， 4000-400 cm'Hresolution 4 cm—1)에서 치환기를 확인하였다 (도 1).
Figure imgf000012_0001
Oxidized hyaluronic acid was analyzed using an FT-IR spectrometer (Cary 640, Agilent Technologies, USA), and the substituent was confirmed at 4000-400 cm ' Hresolution 4 cm- 1 (FIG. 1).
히알루론산의 산화도를 확인하기 위해 17.5 g 의 하이드톡실아민 하이드로클로라이드와 0.05% 메틸 오렌지 6 ι 을 994 m의 증류수에 흔합하여 0.25 M 하이드록실아민 하이드로클로라이드 용액을 만들고 pH 를 4 로 적정하였다. 산화 히알루론산 0.1 g 을 25 의 상기 에 녹이고ᅳ 0.1 mM 수산화나트륨을 이용하여 다시 pH 4 로 적정하였다. 산화도 (%)는 다음의 식을 이용하여 계산하였으며， 결과는 하기 표 1 내지 4와 같았다. 수학식 1  In order to confirm the oxidation degree of hyaluronic acid, 17.5 g of hydroxylamine hydrochloride and 0.05% methyl orange 6 ι were mixed in 994 m of distilled water to make a 0.25 M hydroxylamine hydrochloride solution and the pH was adjusted to 4. 0.1 g of hyaluronic acid oxide was dissolved in 25 of the above, and titrated again to pH 4 using 0.1 mM sodium hydroxide. Oxidation degree (%) was calculated using the following formula, the results were as shown in Tables 1 to 4. Equation 1
¾ -화 i (%) =¾-i = (%)
Figure imgf000012_0002
Figure imgf000012_0002
수^화나 λίίί의농 : 수산하나?쉐의 V ] Χ10~3 1 Λη ^ 화 농 ί 농 농 Χ Χ Χ 10 ~ 3 1 Λη
산화히 , ' 산의^게 ᅳ 腳 Oxidized, ' acidic crab 腳 腳
'.' ';· VH ( repeating unit)의양 【표 1】 M.?. M증량 산화채농도 반믐몬도 반 시간'.' Amount of repeating unit (VH) Table 1 M.?. M increase oxidized vegetable concentration half 믐 Mondo half time
(kDa) (6) (ιΜ) (°0 W간) (kDa) (6) (ιΜ) (° 0 W)
67,4 3.1 15, δ 40 24 67,4 3.1 15, δ 40 24
7 75,9 3,1 15,7 40 24 7 75,9 3,1 15,7 40 24
.79,3 1,0 15,8 40 24  .79,3 1,0 15,8 40 24
【표 2】 Table 2
Figure imgf000013_0001
Figure imgf000013_0001
【표 3】
Figure imgf000014_0001
Table 3
Figure imgf000014_0001
3.3 1,1 2,8 40 3 3.3 1,1 2,8 40 3
3,9 1,0 2.6 RT, 243,9 1,0 2.6 RT, 24
7,6 1,1 2.6 RT, 67,6 1,1 2.6 RT, 6
9.2 1,0 3.8 RT, 69.2 1,0 3.8 RT, 6
9.7 1.0 2.7 40 69.7 1.0 2.7 40 6
9,9 1,0 5,3 RT, 69,9 1,0 5,3 RT, 6
11,3 1,0 3.8 RT. 611,3 1,0 3.8 RT. 6
!4,0 1,0 5.2 RT, 6! 4,0 1,0 5.2 RT, 6
14.3 1,0 2.6 40 2414.3 1,0 2.6 40 24
14.8 3.1 8.0 40 614.8 3.1 8.0 40 6
15,4 1,0 7,8 RL ■6 'ύ15,4 1,0 7,8 RL ■ 6 ' ύ
16.5 1.0 3.8 RT, 616.5 1.0 3.8 RT, 6
17.4 3.1 7.9 40 24 ¾17.4 3.1 7.9 40 24 ¾
1 00 17.7 1.0 5.3 RT. 6 1 00 17.7 1.0 5.3 RT. 6
18,0 1,0 9.6 RT, 6 '"' 18,0 1,0 9.6 RT, 6 '"'
19,5 1,0' 11,1 RT. 619,5 1,0 '11,1 RT. 6
21.0 1,0 11,0 RT. 621.0 1,0 11,0 RT. 6
21,6 1,0 12.7 RT, 621,6 1,0 12.7 RT, 6
21,9 1,0 9,4 RT, 6 '-^21,9 1,0 9,4 RT, 6 '-^
22.8 1.0 7.8 RT. 6 '： S22.8 1.0 7.8 RT. 6 ': S
23.2 1.1 7.9 RT. 6 '：:23.2 1.1 7.9 RT. 6 ' ：
23.6 1.0 12.5 RT, 6 :23.6 1.0 12.5 RT, 6:
24.7 1,0 11.1 RT, 6 . 24.7 1,0 11.1 RT, 6 .
27.0 1.0 9.5 RT, 6 -27.0 1.0 9.5 RT, 6-
30.9 1,0 12,7 RT, 630.9 1,0 12,7 RT, 6
40.8 8,1 15.8 40 2440.8 8,1 15.8 40 24
47.7 3.1 23.5 40 2447.7 3.1 23.5 40 24
82.6 l.l 15.7 40 24 82.6 l.l 15.7 40 24
【표 4】
Figure imgf000014_0002
Table 4
Figure imgf000014_0002
4.6 1.1 2.6 RT. 6 4.6 1.1 2.6 RT. 6
4,7 1,1 2.5 40 64,7 1,1 2.5 40 6
3000 19.8 3,0 7.9 40 24 3000 19.8 3,0 7.9 40 24
23.3 1.1 2,6 40 2 23.3 1.1 2,6 40 2
43.4 3,0 15.9 40 2443.4 3,0 15.9 40 24
,82.2 1.1 15,7 40 24 (2) 2 이상의 아미노기를 갖는 제 2성분 , 82.2 1.1 15,7 40 24 (2) a second component having two or more amino groups
다양한 아미노기 함유 폴리아민 중에서 대표적으로 키토산， 프로타민， PEI, 폴리리신, 스퍼민， 스퍼미딘 및 알부민 등올 제 2 성분으로 이용하였고, 5 중량 % 이상의 폴리아민 수용액으로부터 pH 조절제 (산， 산염， 염기， 염기염 등)를 이용하여 pH 를 8.5， 9.0, 9.5 및 10 으로 조절한 후 상기 산화 히알루론산 /산화 콘드로이친 설페이트의 경우와 동일하게， 동결 건조 후에 수득된 분말을 사용했다.  Among the various amino group-containing polyamines, chitosan, protamine, PEI, polylysine, spermine, spermidine, and albumin were used as second components, and pH adjusters (acid, acid, base, base salt, etc.) from 5% by weight or more of aqueous polyamine solution After adjusting the pH to 8.5, 9.0, 9.5 and 10 using the same as in the case of the oxidized hyaluronic acid / oxidized chondroitin sulfate, the powder obtained after freeze drying was used.
상기 나열된 아미노기 함유 폴리아민의 겔화도와 겔화 시간을 평가하였다. 그 결과， 알부민， 염기성 폴리리신 (BPL), PEI 가 겔화 속도와 젤의 안정성 면에서 우수하였다 (도 2). 이 중에서 BPL 로 하기 실험을 진행하였다. 실시예 2. 물성 평가  The gelation degree and gelation time of the amino group-containing polyamines listed above were evaluated. As a result, albumin, basic polylysine (BPL) and PEI were excellent in terms of gelation rate and gel stability (FIG. 2). Of these, the following experiment was conducted with BPL. Example 2. Evaluation of Physical Properties
(1) 겔화 평가  (1) gelation evaluation
실시예 1 에서 얻은 제 1 및 제 2 성분을 서로 다른 중량비 (1:1, 2:1, The first and second components obtained in Example 1 were mixed in different weight ratios (1: 1, 2: 1,
4:1， 8:1)로 흔합하였다. 흔합한 성분에 물을 뿌려 겔화되는 정도를 확인하였다. 4: 1, 8: 1). The degree of gelation was confirmed by spraying water on the mixed components.
그 결과， 제 1 성분과 제 2 성분을 8:1 로 혼합한 경우에는 10 분 후에 일부 액체로 변하였고， 2:1 과 4:1 로 흔합한 경우는 겔의 탄성이 1:1 에 비하여 비교적 떨어졌다 (도 3). 또한， 3,000 kDa 분자량의 히알루론산을 이용한 흔합제제는 겔화가 되었지만 탄성이 떨어졌다. 분자량 150 kDa 과 1,400 kDa 의 히알루론산의 경우에는 치환도에 상관없이 겔화가 되었으나， 치환도 1 。대 (10-19%)일 때의 흔합제제가 짧은 겔화 시간을 나타냈으며 , 탄성 또한 우수하였다.  As a result, when the first component and the second component were mixed at 8: 1, the liquid turned into some liquid after 10 minutes, and when the mixture was 2: 1 and 4: 1, the elasticity of the gel was relatively higher than 1: 1. Fell (Figure 3). In addition, the mixed agent using hyaluronic acid having a molecular weight of 3,000 kDa gelled but was inelastic. In the case of hyaluronic acid having a molecular weight of 150 kDa and 1,400 kDa, gelation was performed regardless of the degree of substitution. However, when the degree of substitution was 1。 (10-19%), the mixture showed a short gelation time and excellent elasticity.
이상의 결과를 바탕으로， 게 1 성분 (분자량 150 kDa 또는 1,400 kDa의 히알루론산에 알데히드기가 도입된 치환도 의 산화 히알루론산)과 제 2 성분을 1:1의 중량비로 흔합하여 하기의 실험을 실시하였다.  Based on the above results, the following experiment was carried out by mixing one crab component (oxidation hyaluronic acid having a degree of substitution in which an aldehyde group was introduced into a hyaluronic acid having a molecular weight of 150 kDa or 1,400 kDa) and a second component in a weight ratio of 1: 1. .
(2) 겔화 시간 평가 (2) gelation time evaluation
2 ml 류브에 흔합한 성분을 30 mg 씩 소분하여 튜브 뚜정에 모았다. 여기에 120 의 물을 1 초 이내에 뿌린 뒤 겔화되는 시간을 측정하였다. LYDEX 의 경우에는 히알루론산 혼합제제에 비해 적은 양 (80 의 물을 적용했음에도 불구하고 굳어진 젤을 얻는 데에 10 초 이상이 소요되었다. 반면, 산화 히알루론산의 혼합제제의 경우에는 2-3 초 이내의 짧은 겔화 시간을 나타내었다 (도 4 및 표 5 ) . 30 mg each of the ingredients mixed in 2 ml leucine were collected in a tube tablet. 120 hours of water was added to it within 1 second, and time to gelate was measured. In the case of LYDEX, it took more than 10 seconds to obtain a hardened gel despite the use of a smaller amount (80 water applied) compared to the hyaluronic acid mixture. The short gel time of was shown (FIG. 4 and Table 5).
【표 5】  Table 5
Figure imgf000016_0001
Figure imgf000016_0001
(3) 점착력 평가 (3) adhesive force evaluation
100 nig 의 혼합한 성분에 800 ^의 물을 뿌린 후에 Texture Ana lyzer 를 이용하여 점착력을 측정하였다. 그 결과 , LYDEX 는 산화 히알루론산 흔합제제에 비해 적은 양 ( 500 의 물을 적용했음에도 불구하고 평균 점착력이 약 53 .6 gf 로 나타났다. 반면, 산화 히알루론산의 흔합제제의 경우에는 평균 점착력이 각각 약 65.9 및 67.5 gf 로 측정되었다 (도 5 및 표 6) .  After spraying 800 ^ water on the mixed components of 100 nig, the adhesive strength was measured using the Texture Analyzer. As a result, LYDEX showed an average adhesive strength of 53.6 gf even though a small amount (500 water was applied) of LYDEX compared to the oxidized hyaluronic acid mixture. Measured at 65.9 and 67.5 gf (FIG. 5 and Table 6).
【표 6】  Table 6
Figure imgf000016_0002
Figure imgf000016_0002
(4) 흡수력 평가 (4) absorbency evaluation
LYDEX 와， 제 2 성분과 흔합한 각각의 CHO-HA 150 kDa(DS 10%) , CH0- HA 1 , 400 kDa(DS 10%)의 흡수력을 평가하였다. 각 샘플 30 nig 을 페트리 디쉬 (패 60)에 올려놓고 무게를 측정하였다. 37 °C로 미리 데워놓은 증류수를 제품의 흡수력을 고려하여 샘플 무게의 30 배 (30 g)를 첨가하였다. 37 °C의 항온기에서 30 분 동안 방치한 후 페트리 디쉬를 30 초 동안 뒤집어 놓고 그 무게를 측정하였다. 다음의 계산식을 이용하여 흡수력을 계산하였다. 수학식 2 The absorption powers of LYDEX, CHO-HA 150 kDa (DS 10%), CH0-HA 1, and 400 kDa (DS 10%), which were combined with the second component, were evaluated. 30 nig of each sample was placed in a Petri dish (L 60) and weighed. Distilled water preheated to 37 ° C. was added 30 times (30 g) of the sample weight, taking into account the absorbency of the product. After standing in a thermostat at 37 ° C for 30 minutes, the Petri dish was turned upside down for 30 seconds and weighed. Absorption capacity was calculated using the following formula. Equation 2
30붇후 -¾¾ (mg) ~초기투계 (mg)  After 30--¾¾ (mg) to initial permeability (mg)
* 력 (%) = X 100  * Force (%) = X 100
초기부게 (nig)  Early stage (nig)
그 결과， 기존 LYDEX 제형에 비하여 산화 히알루론산의 흔합제제 (CH0-HA ,,400 kDa)의 흡수력이 약 5 배 정도 우수함을 확인하였다 (표 7)·  As a result, it was confirmed that the absorption power of the mixed agent of oxidized hyaluronic acid (CH0-HA ,, 400 kDa) was about 5 times better than the conventional LYDEX formulation (Table 7).
【표 7】  Table 7
Figure imgf000017_0001
실시예 3. 생체 내 평가
Figure imgf000017_0001
Example 3. In Vivo Evaluation
(1) 실험동물  (1) experimental animals
체중 2 내지 3 kg 의 3 마리의 수컷 토끼 (New Zealand White; 오리엔트 바이오 (Orient Bio), 성남， 대한민국)를 실험에 사용하였다. 모든 동물 사육 및 시험 과정은 인하대학교의 실험동물연구위원회 (Experimental Animal Research Co瞧 ittee)의 지침에 따라 수행하였다.  Three male rabbits weighing 2-3 kg (New Zealand White; Orient Bio, Seongnam, South Korea) were used for the experiment. All animal breeding and testing procedures were conducted according to the guidelines of the Experimental Animal Research Co. Ittee of Inha University.
(2) 위 출혈 유발 동물 모델 (2) gastric bleeding-induced animal model
토끼의 점막절제 (mucosectomy)-유도 위출혈 모델을 다음과 같이 제작하였다. 수술 전 24 시간 동안 토끼를 절식시키고， 케타민 (4.2 mg/kg) 및 실라진 (11.7 mg/kg)의 흔합물을 근육 주사하여 마취시켰다. 토끼의 상복부를 절개하여 위를 노출시키고， 위의 대만곡 (greater curvature)을 따라 약 5-7 cm 정도 절개하였다. 등장 생리식염수 200 를 위의 점막하 층 (submucosal layer)에 주사하고， 팽윤된 위 점막을 수술가위를 사용하여 절제하였다. 절제된 부위의 직경은 약 7-10瞧 이었다. (3) 점막 부착능력 및 지혈능력 평가  A mucosectomy-induced gastric bleeding model in rabbits was constructed as follows. The rabbits were fasted for 24 hours prior to surgery and anesthetized by intramuscular injection of a combination of ketamine (4.2 mg / kg) and silazine (11.7 mg / kg). The upper abdomen of the rabbit was incised to expose the stomach and incision about 5-7 cm along the great curvature of the stomach. Isotonic saline 200 was injected into the submucosal layer of the stomach, and the swollen gastric mucosa was excised using surgical scissors. The diameter of the excised site was about 7-10 mm 3. (3) Evaluation of mucoadhesive and hemostatic capacity
출혈이 되고 있는 절제된 토끼의 위점막에 약 0.5 g 의 흔합제제 (1:1 중량비의 제 1 성분와 제 2성분의 흔합물)를 도포하였다. 그 결과， 도 6 에 나타난 바와 같이， 흔합제제를 도포한 즉시 흔합제제와 혈액이 반응하여 흔합제제의 겔화가 이루어졌고 비처치군에 비해 출혈시간이 단축되었다. 또한， 본 발명의 조성물의 점막 부착능력을 확인하였다 (도 6) . 실시예 4. 의료용 접착제의 제조 ② About 0.5 g of a combination agent (a mixture of the first component and the second component in a 1: 1 weight ratio) was applied to the gastric mucosa of the bleeding rabbit. As a result, in FIG. As shown, as soon as the mixture was applied, the mixture and the blood reacted to gel the mixture and shorten the bleeding time compared to the untreated group. In addition, the mucoadhesive capacity of the composition of the present invention was confirmed (FIG. 6). Example 4 Preparation of Medical Adhesive
(1) 산화 히알루론산과산화콘드로이친 설페이트 (제 1 성분)의 제조 분자량 1 , 400 kDa 의 히알루론산 (SHANDONG BL00MAGE FREDA BIOPHARM CO . , Ltd) 3 g 을 150 의 증류수에 용해시켰다. 다음에 표 1 과 같이 과요오드산 나트륨 (분자량 213.89)을 첨가하고， 반응 플라스크를 40°C에서 24 시간 교반하면서 반웅시켰다. 그리고， 반웅 후의 용액을 증류수로 48 시간 투석 (분획분자량 12000-14000 의 투석막 사용)한 후 동결 건조하였다. (1) Preparation of oxidized hyaluronic acid and chondroitin peroxide sulfate (first component) 3 g of hyaluronic acid (SHANDONG BL00MAGE FREDA BIOPHARM CO., Ltd) having a molecular weight of 1,400 kDa was dissolved in 150 distilled water. Next, sodium periodate (molecular weight 213.89) was added as shown in Table 1, and the reaction flask was reacted with stirring at 40 ° C for 24 hours. After the reaction, the solution was dialyzed with distilled water for 48 hours (using a dialysis membrane having a molecular weight of 12000-14000), and then freeze-dried.
분자량 5， 000-50， 000 사이의 콘드로이친 설페이트 (YANTAI D0NGCHENG BIOCHEMICAL CO. , Ltd) 3 g을 15 mi의 증류수에 용해시켰다. 다음에 표 2 과 같이 과요오드산 나트륨 (분자량 213.89)을 첨가하고, 상온에서 18 시간 교반하면서 반응시켰다. 그리고， 반응 후의 용액을 증류수로 48 시간 투석 (분획분자량 12000-14000의 투석막사용)한 후 동결 건조하였다.  3 g of Chondroitin Sulfate (YANTAI D0NGCHENG BIOCHEMICAL CO., Ltd) having a molecular weight of 5, 000-50, 000 were dissolved in 15 mi of distilled water. Next, sodium periodate (molecular weight 213.89) was added as shown in Table 2, and the reaction was stirred at room temperature for 18 hours. The solution after the reaction was dialyzed with distilled water for 48 hours (using a dialysis membrane of fractional molecular weight 12000-14000), and then freeze-dried.
이하의 실험에서는 산화 히알루론산과 산화 콘드로이친 설페이트를 제 1 성분으로서 사용했다.  In the following experiment, oxidized hyaluronic acid and chondroitin oxide were used as the first component.
NaOH 적정법으로 히알루론산과 콘드로이친 설페이트의 치환도 (산화도)를 확인하였다. 구체적으로는， 17.5 g 의 하이드록실아민 하이드로클로라이드와 0.05% 메틸 오렌지 6 11 을 994 ^의 증류수에 혼합하여 0.25 1∞1/ £의 하이드록실아민 하이드로클로라이드 용액을 만들고 pH를 4로 적정하였다. 산화 히알루론산 흑은 콘드로이친 설페이트 0. 1 g을 25 m의 상기 용액에 녹이고, 0. 1 \/ ί 수산화나트륨을 이용하여 다시 pH 4 로 적정하였다. 치환도 (%)는 다음의 식을 이용하여 계산하였으며， 결과는 하기 표 8 및 9와 같았다. 수학식 3
Figure imgf000019_0001
The degree of substitution (oxidation degree) of hyaluronic acid and chondroitin sulfate was confirmed by NaOH titration. Specifically, 17.5 g of hydroxylamine hydrochloride and 0.05% methyl orange 6 11 were mixed in 994 ^ distilled water to make a 0.25 1∞1 / £ hydroxylamine hydrochloride solution and the pH was titrated to 4. 0.1 g of hyaluronic acid black silver chondroitin sulfate was dissolved in 25 m of the above solution and titrated again to pH 4 with 0.1 \ / ί sodium hydroxide. Substitution degree (%) was calculated using the following formula, and the results were as shown in Tables 8 and 9. Equation 3
Figure imgf000019_0001
수산화니 의농도 X수 ¾화나트 f의年피 > (r3 Concentration of nitric hydroxide x number ¾ sulphate f> (r 3
ᅳ 산하 미노: ¾리^의무? ό ᅳ  미 Affiliated Mino: What are your obligations? ό ᅳ
굴리. . re ting (init).s|  Guri. . re ting (init) .s |
【표 8】 Table 8
Figure imgf000019_0002
Figure imgf000019_0002
【표 9]  [Table 9]
Figure imgf000019_0003
Figure imgf000019_0003
(2) 2 이상의 아미노기를 갖는 제 2성분의 제조 (2) Preparation of Second Component Having Two or More Amino Groups
다양한 아미노기 함유 폴리아민 중에서 대표적으로 키토산, 프로타민 PEI , 폴리리신， 스퍼민， 스퍼미딘 및 알부민 등을 제 2 성분으로 선택하였고, 제 2 성분으로서는 pH 에 따른 겔화 여부를 알아보기 위하여 수용액 상태에서 pH 를 여러 범위 (5.5-6.4， 6.5-7.4, 7.5-8.4, 8.5-9.4， 9.5-10.4, 10.5-11)로 조절한 후, 상기 산화 히알루론산 /산화 콘드로이친 설페이트의 경우와 동일하게， 동결 건조 후에 수득된 분말을 사용하였다. 상기 제 1 성분의 산화 히알루론산 /산화 콘드로이친 설페이트와 폴리아민을 흔합하였다. 그 결과, 폴리아민의 종류에 관계없이 pH
Figure imgf000020_0001
8.5-11 일 경우에만 겔화됨을 확인하였다. 일예로 폴리 -L-리신의 pH 가 8.5 인 경우에는 젤이 형성되었지만， pH 가 5.6 에서는 젤이 형성되지 않았다. 본 실험에서 젤의 형성 유무는 젤의 투명도에 의해서 결정하였다 (투명: 젤화， 불투명: 젤화되지 않음) (도 7) . 실시예 5. 최적의 비율 확인 Among the various amino group-containing polyamines, chitosan, protamine PEI, polylysine, spermine, spermidine, and albumin were selected as the second component, and as a second component, to find out whether the gelation was performed according to pH. In aqueous solution, after adjusting the pH to various ranges (5.5-6.4, 6.5-7.4, 7.5-8.4, 8.5-9.4, 9.5-10.4, 10.5-11), the same as in the case of the oxidized hyaluronic acid / oxidized chondroitin sulfate The powder obtained after freeze drying was used. Oxidized hyaluronic acid / oxidized chondroitin sulfate of the first component and polyamine were mixed. As a result, regardless of the type of polyamine
Figure imgf000020_0001
It was confirmed that the gelation only in the case of 8.5-11. For example, a gel was formed when the pH of poly-L-lysine was 8.5, but no gel was formed when the pH was 5.6. Gel formation in this experiment was determined by the transparency of the gel (transparency: gelling, opacity: not gelling) (FIG. 7). Example 5. Confirmation of Optimal Ratio
(1) 분자량， 비율별 최적 조건 확립  (1) Establish optimum conditions for molecular weight and ratio
수득된 분말상의 산화 콘드로이틴 설페이트와 분자량 150-3 , 000 kDa 인 산화 히알루론산 (1 : 1)을 산화도에 따라 비율별로 폴리아민 (PA; pH 8.5-9.5 폴리리신 선정하여 사용)과 섞어 물성을 확인하였다. 산화도에 따라 비율별로 섞은 분말 50 mg 에 멸균된 증류수 200 ^를 넣어 멸균된 증류수를 홉수하는지 육안으로 그 정도를 확인하였다. 분말이 멸균된 증류수를 10 초 이내 흡수하기 시작 할 때를 좋음 (+++) , 30 초 이내를 보통 (++) , 60 초 이상올 나쁨 (+)으로 평가하여 용해도 (solubi l i ty)를 확인하였다. 또한, 멸균된 증류수를 넣었을 때 겔화가 되는지 확인하고 그 시간을 측정하였다. 이렇게 만들어진 겔이 다시 액화 되는지 확인하여 그 시간을 측정하였다. 이렇게 확인한 결과를 하기 표에 정리하여 나타내었다 (표 10 및 11) .  The physical properties of the obtained powdered chondroitin sulfate and molecular weight 150-3, 000 kDa phosphorus oxidized hyaluronic acid (1: 1) were mixed with polyamine (PA; selected by pH 8.5-9.5 polylysine) in proportion to the degree of oxidation. It was. Sterilized distilled water 200 ^ was added to 50 mg of the powder mixed by ratio according to the degree of oxidation, and the degree was visually checked to see if the sterilized distilled water was hopped. When the powder starts to absorb sterilized distilled water within 10 seconds, the solubility (solubi li ty) is evaluated as good (+++), within 30 seconds as normal (++), and as bad (+) for more than 60 seconds. Confirmed. In addition, when the sterilized distilled water was added to determine whether the gelation and the time was measured. This gel was confirmed to be liquefied again and the time was measured. The results thus confirmed are summarized in the following table (Tables 10 and 11).
표 11 에 나타난 바와 같이， 거의 모든 제 1 성분과 제 2 성분의 조합에서 수분흡수력과 겔화 시간에 있어 만족할만한 결과가 도출되었으며， 이중에서 최상의 성능 (수분흡수력이 +++， 겔화 시간이 30 초 이내)을 보인 것은 10-50% 산화 콘드로이틴 설페이트와 10-40% 산화 히알루론산 (150， 1 ,400 또는 3 ,000 kDa)을 폴리아민과 1 : 1 의 비율로 섞은 흔합제제였으며 (샘플번호: 3, 4, 12， 14, 17, 26, 29)， 이 결과를 바탕으로 하기 실시예에서는 #14(UI-SAH 명명)를 사용하여 실험을 진행하였다. 【표 10] As shown in Table 11, satisfactory results in water absorption and gelation time were obtained for almost all combinations of the first and second components, of which the best performance (water absorption capacity is +++ and gelation time is 30 seconds). ) Was a mixture of 10-50% oxidized chondroitin sulfate and 10-40% oxidized hyaluronic acid (150, 1,400 or 3,000 kDa) with polyamine in a ratio of 1: 1 (sample number: 3) , 4, 12, 14, 17, 26, 29), and based on this result, the experiment was conducted using Example 14 (named UI-SAH). Table 10
Figure imgf000021_0001
Figure imgf000021_0001
Figure imgf000022_0001
Figure imgf000022_0001
(1) 간엽 절제술 모델 (1) Mesenchymal resection model
체중 200-300 g 의 수컷 SD 래트의 복강에 케타민과 럼푼 흔합물을 주사하여 마취한 후 중앙 상복부를 세로 또는 가로로 약 3-4 cm 정도 절개하였다. 절개된 복부의 틈으로 젖은 거즈를 이용하여 간엽을 노출시키고, 간문맥 및 간동맥를 혈관용 클립으로 결찰하였다. 간엽의 가장자리에서 약 1 cm 정도 떨어진 곳을 수술용 가위를 이용하여 절제하고， UI-SAH 50-100 mg 를 도포하였다. 대조군으로는 Ar i sta™ AH(Medafor Inc . , USA)를 도포하였다. 도포 후 결찰 하였던 클립을 제거한 후 출혈이 발생하는지 확인하고， 멸균된 거즈를 이용하여 출혈량을 측정하였다.  Anesthesia was injected by injection of a ketamine and lump mixture into the abdominal cavity of male SD rats weighing 200-300 g, and the central upper abdomen was incised approximately 3-4 cm in length or width. The mesenchyme was exposed using a wet gauze into the incision of the incised abdomen, and the portal vein and hepatic artery were ligated with a vascular clip. About 1 cm away from the edge of the mesenchyme was excised with surgical scissors and 50-100 mg of UI-SAH was applied. As a control, Ar i sta ™ AH (Medafor Inc., USA) was applied. After application, the ligated clip was removed after ligating, and the amount of bleeding was measured using sterile gauze.
(2) 신장 절제술 모델 (2) nephrectomy model
체중 200-300 g 의 수컷 SD 래트의 복강에 케타민과 럼푼 흔합물을 주사하여 마취한 후 우측 복부를 세로로 약 3-4 cm 정도 절개하였다. 절개된 복부의 틈으로 젖은 거즈를 이용하여 신장을 노출시키고， 신정맥 (renal vein) , 신동맥 (renal arter i es)올 혈관용 클립으로 결찰하였다. 신장의 가장자리에서 약 1 cm 정도 떨어진 곳을 수술용 가위를 이용하여 절제하고， UI— SAH 50-100 mg 을 도포하였다. 1 대조군으로는 Ar i sta™ AH(Medafor Inc . , USA)를 도포하였다. 도포 후 결찰 하였던 클립을 제거 한 후 출혈이 발생하는지 확인하고, 멸균된 거즈를 이용하여 출혈량을 측정하였다. Anesthetized by injection of a ketamine and lump mixture into the abdominal cavity of male SD rats weighing 200-300 g, the right abdomen was incised approximately 3-4 cm in length. Renal veins were exposed to the resected abdominal gap with wet gauze and ligated with clips for renal veins and renal arteries. About 1 cm from the edge of the kidney was removed with surgical scissors and UI—SAH 50-100 mg was applied. As a control, Ar i sta ™ AH (Medafor Inc., USA) was applied. After application, the ligated clip was removed after ligating, and the amount of bleeding was measured using sterile gauze.
(3) 위점막 절제술 모델 (3) gastric mucosal resection model
체중 200-300 g 의 수컷 SD 래트를 24 시간 절식시킨 후 복강에 케타민과 럼푼 흔합물을 주사하여 마취한 후 중앙 상복부를 세로 또는 가로로 약 3-4 cm 정도 절개하였다. 절개된 복부의 름으로 젖은 거즈를 이용하여 위를 노출시키고 혈관이 적은 위의 만곡된 부위를 수평으로 약 3 cm 정도 절개하여 위 내벽이 보이게 하였다. 위 내벽에 주사용 생리식염수 100 를 주입한 후 위점막을 수술용 가위로 약 5 mm 직경의 원이 되도록 절제하고 UI-SAH 50-100 mg 을 도포하였다. 대조군으로는 Ar i sta™ AH(Medafor Inc . , USA)를 도포하였다. 도포 부위에 출혈이 발생하는지 확인하고 멸균된 거즈를 이용하여 출혈량을 측정하였다. (4) 혈관 (Portal vein) 출혈 모델 Male SD rats weighing 200-300 g were fasted for 24 hours, and then anesthetized by injecting a ketamine and lump mixture into the abdominal cavity, and the central upper abdomen was incised about 3-4 cm vertically or horizontally. The stomach was exposed using a wet gauze to the incision of the incised abdomen, and the inner wall of the stomach was made visible by cutting about 3 cm horizontally in the curved area of the stomach where there were few blood vessels. After injection of 100 physiological saline into the stomach lining, the gastric mucosa was excised to a circle of about 5 mm diameter with surgical scissors and 50-100 mg of UI-SAH was applied. As a control, Ar i sta ™ AH (Medafor Inc., USA) was applied. The bleeding occurred at the application site and the amount of bleeding was measured using sterile gauze. (4) Portal vein bleeding model
체중 200-300 g 의 수컷 SD 래트의 복강에 케타민과 럼푼 흔합물을 주사하여 마취한 후 중앙 상복부를 세로 또는 가로로 약 5-6 cm 정도 절개하였다. 절개된 복부의 틈으로 다른 장기들을 좌축으로 이동시킨 후 문맥 (Portal vein)을 노출시켰다. 문맥의 위와 아래 두 곳을 혈관용 클립으로 결찰하였다. 18 게이지 바늘을 이용하여 문맥에 구멍을 낸 후 UI- SAH 50-100 mg 을 도포하였다. 대조군으로는 Ar i sta™ AH(Medafor Inc . , USA)를 도포하였다. 도포 후 결찰하였던 클립을 제거 한 후 출혈이 발생하는지 확인하고, 멸균된 거즈를 이용하여 출혈량올 측정하였다.  Anesthesia was injected by injection of a ketamine and lump mixture into the abdominal cavity of male SD rats weighing 200-300 g, and the central upper abdomen was incised about 5-6 cm vertically or horizontally. The portal vein was exposed after moving other organs to the left axis through the incised abdominal gap. Two sites above and below the portal vein were ligated with a vascular clip. UI-SAH 50-100 mg was applied after puncturing the portal vein using an 18 gauge needle. As a control, Ar i sta ™ AH (Medafor Inc., USA) was applied. After application, the ligated clip was removed after removing the ligated clip, and the bleeding amount was measured using sterile gauze.
(5) 실험결과 (5) Experiment result
실험결과는 도 8 내지 12에 나타내었다. 도 12에 나타난 바와 같이 , 본 발명의 조성물의 지혈효과는 대조군 (Ar i sta™ AH) 보다 더 우수하였다. 이상으로 본 발명의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현 예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항과 그와 등가물에 의하여 정의된다고 할 것이다.  Experimental results are shown in FIGS. 8 to 12. As shown in FIG. 12, the hemostatic effect of the composition of the present invention was better than that of the control group (Ar i sta ™ AH). Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims

【특허청구범위】 【Patent Claims】
【청구항 1】 【Claim 1】
다음을 포함하는 생분해성 의료용 접착제 또는 실란트 조성물: A biodegradable medical adhesive or sealant composition comprising:
(a) 포밀기가 도입되어 산화된 산화 글리코사미노글리칸을 포함하는 제 1 성분; 및 (a) a first component comprising oxidized glycosaminoglycan oxidized by introducing a formyl group; and
(b) 2 이상의 아미노기를 갖는 폴리아민을 포함하는 제 2 성분， 상기 제 2 성분의 수용액 상태에서의 pH는 8.5 내지 11.0임 . (b) A second component containing a polyamine having two or more amino groups, and the pH of the second component in an aqueous solution is 8.5 to 11.0.
【청구항 2] [Claim 2]
제 1 항에 있어서, 상기 산화 글리코사미노글리칸은 산화 히알루론산， 산화 콘드로이틴 설페이트， 산화 콘드로이틴， 산화 더마탄 설페이트, 산화 헤파란 설페이트, 산화 헤파린 및 산화 케라탄 설페이트로 구성된 군으로부터 선택되는 것을 특징으로 하는 조성물. The method of claim 1, wherein the oxidized glycosaminoglycan is selected from the group consisting of oxidized hyaluronic acid, oxidized chondroitin sulfate, oxidized chondroitin, oxidized dermatan sulfate, oxidized heparan sulfate, oxidized heparin, and oxidized keratan sulfate. A composition made of.
【청구항 3】 【Claim 3】
제 1 항에 있어서， 상기 산화 글리코사미노글리칸은 시 -히 = ¾^ χ 100 The method of claim 1, wherein the oxidized glycosaminoglycan is si -hi = ¾^ χ 100
' ' ᅳ 산회. ; 리.코 미^ ' 에 따라 계산한 산화도 (%)가 10 내지 99.5%인 것을 특징으로 하는 조성물. '' ᅳ Sanhoe. ; A composition characterized in that the degree of oxidation ( % ) calculated according to Li.comi^ ' is 10 to 99.5%.
【청구항 4】 【Claim 4】
제 1 항에 있어서, 상기 제 1 성분은 2 종류 이상의 산화 글리코사미노글리칸을 포함하는 것을 특징으로 하는 조성물. The composition according to claim 1, wherein the first component contains two or more types of oxidized glycosaminoglycans.
【청구항 5】 【Claim 5】
제 4 항에 있어서, 상기 2 종류 이상의 산화 글리코사미노글리칸은 산화 히알루론산 및 산화 콘드로이틴 설페이트인 것을 특징으로 하는 조성물. The composition according to claim 4, wherein the two or more types of oxidized glycosaminoglycans are oxidized hyaluronic acid and oxidized chondroitin sulfate.
【청구항 6】 【Claim 6】
제 5 항에 있어서, 상기 산화 히알루론산은 10-40%의 산화도를 가지고， 상기 산화 콘드로이틴 설페이트는 10-55%의 산화도를 갖는 것을 특징으로 하는 조성물. The composition of claim 5, wherein the oxidized hyaluronic acid has an oxidation degree of 10-40%, and the oxidized chondroitin sulfate has an oxidation degree of 10-55%.
【청구항 7】 【Claim 7】
제 1 항에 있어서， 상기 폴리아민은 폴리리신， 키토산， 알부민， 푸트레신 (putrescine) , 가다베린 (cadaver ine)， 스페르미딘 ( spermidine)， 스페르민 (spermine) , 프로타민 및 PEI (Polyethyl enimine)로 구성된 군으로부터 선택된 조성물. The method of claim 1, wherein the polyamine is polylysine, chitosan, albumin, putrescine, cadaverine, spermidine, spermine, protamine, and PEI (polyethyl enimine). ) A composition selected from the group consisting of.
【청구항 8] [Claim 8]
제 1 항에 있어서， 상기 의료용 접착제 또는 실란트 조성물은 생체 조직의 접착, 층전， 도포， 유착 방지， 창상 피복 및 지혈로 구성된 군으로부터 선택된 의료 용도를 갖는 것을 특징으로 하는 조성물. The composition according to claim 1, wherein the medical adhesive or sealant composition has a medical use selected from the group consisting of adhesion, layering, application, anti-adhesion, wound covering, and hemostasis of living tissue.
【청구항 9】 【Claim 9】
제 1 항에 있어서， 상기 생분해성 의료용 접착제 또는 실란트 조성물은 아민기를 갖는 약물을 더 포함하는 것을 특징으로 하는 조성물. The composition according to claim 1, wherein the biodegradable medical adhesive or sealant composition further comprises a drug having an amine group.
【청구항 10】 【Claim 10】
제 1 항의 생분해성 의료용 접착제 또는 실란트 조성물을 접착， 충전， 도포， 유착 방지, 창상 피복 또는 지혈이 필요한 생체 조직에 적용하는 단계를 포함하는 생체 조직의 접착ᅳ 층전, 도포, 유착 방지, 창상 피복 또는 지혈방법 . Adhesion of biological tissue, including the step of applying the biodegradable medical adhesive or sealant composition of claim 1 to biological tissue requiring adhesion, filling, application, anti-adhesion, wound covering, or hemostasis - layering, application, anti-adhesion, wound covering, or How to stop bleeding.
PCT/KR2014/012265 2013-12-13 2014-12-12 Biodegradable medical adhesive or sealant composition WO2015088275A1 (en)

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CN201480067775.9A CN106061518B (en) 2013-12-13 2014-12-12 Biodegradable medical adhesive or sealant composition
CA2933271A CA2933271C (en) 2013-12-13 2014-12-12 Biodegradable medical adhesive or sealant composition
US15/102,406 US10105465B2 (en) 2013-12-13 2014-12-12 Biodegradable medical adhesive or sealant composition
JP2016538741A JP6207745B2 (en) 2013-12-13 2014-12-12 Biodegradable medical adhesive or sealant composition
EP14870365.5A EP3081236B1 (en) 2013-12-13 2014-12-12 Biodegradable medical adhesive or sealant composition
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CN116687959A (en) * 2022-05-18 2023-09-05 朱小丰 Chondroitin sulfate biological polyamine complex, and preparation method and application thereof

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