WO2023282583A1 - Agent hémostatique de type poudre comprenant un polymère biocompatible et son procédé de préparation - Google Patents

Agent hémostatique de type poudre comprenant un polymère biocompatible et son procédé de préparation Download PDF

Info

Publication number
WO2023282583A1
WO2023282583A1 PCT/KR2022/009675 KR2022009675W WO2023282583A1 WO 2023282583 A1 WO2023282583 A1 WO 2023282583A1 KR 2022009675 W KR2022009675 W KR 2022009675W WO 2023282583 A1 WO2023282583 A1 WO 2023282583A1
Authority
WO
WIPO (PCT)
Prior art keywords
powder
polymer
hemostatic agent
type
chitosan
Prior art date
Application number
PCT/KR2022/009675
Other languages
English (en)
Korean (ko)
Inventor
권오형
곽동민
Original Assignee
금오공과대학교 산학협력단
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 금오공과대학교 산학협력단 filed Critical 금오공과대학교 산학협력단
Publication of WO2023282583A1 publication Critical patent/WO2023282583A1/fr

Links

Images

Classifications

    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/12—Ketones
    • A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/36—Blood coagulation or fibrinolysis factors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/36—Blood coagulation or fibrinolysis factors
    • A61K38/363—Fibrinogen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/46—Hydrolases (3)
    • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/46—Hydrolases (3)
    • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
    • A61K38/482—Serine endopeptidases (3.4.21)
    • A61K38/4833—Thrombin (3.4.21.5)
    • 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
    • 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
    • A61L24/0015—Medicaments; Biocides
    • 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
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • 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
    • A61L26/0066—Medicaments; Biocides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
    • 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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/418—Agents promoting blood coagulation, blood-clotting agents, embolising agents
    • 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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/428—Vitamins, e.g. tocopherol, riboflavin
    • 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
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/04—Materials for stopping bleeding

Definitions

  • the present invention relates to a powder-type hemostatic agent containing a biocompatible polymer and a method for manufacturing the same, and more specifically, it is characterized by excellent biocompatibility and high blood absorption through a cross-linked structure by using a natural polymer.
  • a hemostatic material by including a hemostatic material, the rate of clot formation is fast, many blood clots are formed, and blood coagulation ability is excellent, and tissue adhesion can be increased by including an adhesive, and an excellent hemostatic effect is provided.
  • Blood is discharged out of the blood vessel due to trauma or internal bleeding that exists within the blood vessel, which is called hemorrhage.
  • hemostasis occurs to prevent blood loss.
  • the hemostasis process is largely classified into primary hemostasis and secondary hemostasis.
  • blood flow is first hindered by vasoconstriction, and platelets adhere to the inner wall of blood vessels with the help of vWF (von Willebrand factor) and aggregate to form the primary hemostasis.
  • vWF von Willebrand factor
  • a platelet plug is formed, which is called primary hemostasis.
  • secondary hemostasis is achieved as insoluble fibrin is produced through continuous activation of various types of blood coagulation factors and finally a secondary hemostatic plug is formed.
  • Hemostasis methods include a mechanical method, a cold/hot method, and a chemical method.
  • the most basic mechanical method is to directly press the bleeding area using gauze or cotton, or to stop bleeding by using instruments such as sutures, clamps, and clips for bleeding caused by cuts or lacerations.
  • bone bleeding bone wax
  • the cold and hot method there is a cold compress method that suppresses blood going to the bleeding area by lowering the body temperature and a method of coagulating the tissue by applying heat to the tissue using an electrocautery machine, which is one of electrosurgical devices using high-frequency electrical energy, to stop bleeding.
  • a typical chemical method includes a hemostatic agent.
  • Hemostatic agents are largely astringents that suppress direct bleeding by an astringent action that forms a film when applied to the skin or mucous membranes, blood coagulation accelerators that promote blood coagulation by using various factors involved in the blood coagulation process as drugs when bleeding occurs, and blood coagulants.
  • Topical hemostats are produced in various forms such as sponge, sheet, hydrogel, bar, and powder. Hemostats in the form of sponges and sheets can be applied to wide wounds and can absorb a large amount of blood, but have the disadvantage of having to be cut to fit the size of the wound and weak adhesiveness. Hydrogel-type hemostatic agents can absorb a large amount of water due to the network structure between polymer chains, can be applied to irregular surfaces, and have excellent adhesiveness, but because they have a lot of moisture, viruses or microorganisms can easily propagate and cause infection. there is.
  • powder type hemostatic agent Since the powder type hemostatic agent has a large surface area compared to other hemostatic agents, the contact area with blood is widened, enabling rapid hemostasis. However, due to the nature of the powder itself, it does not participate in hemostasis and is messy due to the powder around the wound. Powder-type hemostats currently on the market form a film to stop bleeding temporarily and stop bleeding. If the film is removed, bleeding may occur again, and if the amount of blood is large, the film cannot be properly attached to the tissue and is lost along with the blood. has
  • the present invention has been made to solve the problems of the prior art, to provide a powder-type hemostatic agent having excellent biocompatibility and high blood absorption through a crosslinked structure using natural polymers, and a manufacturing method thereof.
  • Another object of the present invention is to provide a powder-type hemostatic agent having a high blood clot formation rate by including a hemostatic substance and having a high clot formation rate.
  • tissue adhesion by including an adhesive, and to provide a powder-type hemostatic agent having an excellent hemostatic effect.
  • first polymer a first polymer
  • second polymer a second polymer
  • cross-linking agent wherein the first polymer and the second polymer are biocompatible polymers, and the first polymer and the second polymer are cross-linked by the cross-linking agent.
  • the biocompatible polymer is chitosan, starch, cellulose, alginate, gelatin, hyaluronic acid, sodium hyaluronate, collagen, polyvinyl alcohol (PVA), alginic acid, pectin, carrageenan, chondroitin (sulfate), dextran (sulfate), polylysine (polylysine), carboxymethyl titin, fibrin, agarose, pullulan, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), hydroxypropylcellulose (HPC), hydroxyethylcellulose (HEC), hydroxypropyl Methylcellulose (HPMC), sodium carboxymethylcellulose, polyalcohol, gum arabic, cyclodextrin, dextrin, glucose, fructose, trehalose, glucose, maltose, lactose, lactulose, fructose, turanose, melitose, melegitose , dex
  • the first polymer may include chitosan
  • the second polymer may include starch
  • the crosslinking agent is glyoxal, glutaraldehye, citric acid, At least one selected from the group consisting of N,N-methylene bisacrylamide, dialdehyde, oxalic acid and citric acid can include
  • the powder-type hemostatic agent may further include at least one selected from the group consisting of an adhesive, a calcium compound, and a hemostatic material.
  • the adhesive may include at least one selected from the group consisting of poly(vinyl pyrrolidone), PVP, pullulan, and dopamine.
  • the amount of the adhesive may be 0.5 to 15 parts by weight relative to 100 parts by weight of the powdered hemostatic agent.
  • the calcium compound may include at least one selected from the group consisting of calcium chloride, calcium nitrate, calcium carbonate, calcium acetate, calcium carbonate, calcium sulfide, calcium carbonate and calcium phosphate.
  • the hemostatic material may include at least one selected from the group consisting of thrombin, prothrombin, thromboplastin, fibrinogen, aprotinin, vitamin C and vitamin K.
  • Another aspect of the present invention (a) preparing a first mixed solution by preparing a first polymer solution and a second polymer solution, respectively, and mixing them; (b) preparing a cross-linked product in which the first polymer and the second polymer are cross-linked by adding a cross-linking agent to the first mixed solution; and (c) lyophilizing the cross-linked product and pulverizing it to prepare a powder-type hemostatic agent.
  • step (b) mixing the crosslinked product with a solution containing at least one selected from the group consisting of an adhesive, a calcium compound and a hemostatic material to prepare a second mixed solution Step; may be further included.
  • the step (c) may be (c') preparing a powder-type hemostatic agent by lyophilizing and pulverizing the second mixed solution.
  • step (c) After freeze-drying in step (c), cooling under nitrogen gas conditions may be further included.
  • the powdered hemostatic agent of the present invention has excellent biocompatibility by using natural polymers and has a high blood absorption rate through a cross-linked structure.
  • the powder-type hemostatic agent of the present invention contains a hemostatic material, so that the rate of clot formation is fast, and many clots are formed, so that the blood coagulability is excellent.
  • the powder-type hemostatic agent of the present invention can increase tissue adhesion by including an adhesive, and has an effect of having an excellent hemostatic effect.
  • 1 is a schematic diagram showing the structure of the powdered hemostatic agent of the present invention.
  • Example 2 is a graph showing the blood absorption rate according to the Chitosan/starch content ratio adjustment of the chitosan/starch powder type hemostatic agent according to Example 1.
  • Figure 3 is a graph showing the blood absorption rate according to the crosslinking time in Example 1, (a) 2 hours, (b) 4 hours, (c) 12 hours, (d) 24 hours, (e) 72 hours.
  • Figure 4 is a graph showing the blood absorption rate according to the crosslinking agent content in Example 1, (a) 1000 ⁇ l, (b) 500 ⁇ l, (c) 100 ⁇ l, (d) 50 ⁇ l, (e) 25 ⁇ l, (f) is 10 ⁇ l.
  • Figure 5 is a graph showing the blood absorption rate according to the particle size of the chitosan / starch powder type hemostat according to Example 1, (a) 250-500 ⁇ m, (b) 125-250 ⁇ m, (c) 0-125 ⁇ m .
  • Figure 6 is the FTIR analysis result, (A) chitosan, (B) starch, (C) FTIR spectrum of the chitosan / starch powder type hemostat prepared according to Example 1.
  • Example 7 is a graph comparing the adhesion of chitosan/starch powder type hemostat (Freeze dry) prepared according to Example 1, Pure PVP and chitosan/starch powder type hemostat containing 15 wt% PVP prepared according to Example 2 .
  • Example 8 is a graph comparing the adhesiveness according to the PVP concentration of chitosan/starch powder-type hemostatic agents containing PVP prepared in Example 2.
  • FIG 9 is a graph showing the blood absorption rate according to the PVP concentration of the chitosan/starch powder-type hemostatic agent containing PVP prepared according to Example 2.
  • 10 is a graph showing fibrin formation time according to CaCl 2 concentration.
  • 11 is a graph showing fibrin formation time according to thrombin concentration.
  • FIGs 12 a1 to i2 (A) is a chitosan/starch powder type hemostat prepared according to Example 1, (B) is a chitosan/starch powder type hemostat containing CaCl 2 /thrombin prepared according to Example 3, ( C) to (I) are SEM images of chitosan/starch powder-type hemostat containing PVP/CaCl 2 /thrombin prepared according to Example 4, and (C) to (I) show PVP concentrations of 0.5, 1, 3, 5, 7, 10, 15 wt%.
  • (A) is a chitosan/starch powder type hemostat prepared according to Example 1
  • (B) is a chitosan/starch powder type hemostat containing CaCl 2 /thrombin prepared according to Example 3 is the result of EDS analysis of
  • Figure 14 is the results of the whole blood coagulation test (Lee-White method) of the powder-type hemostat prepared according to Example 1, Example 3 and Example 4.
  • Figure 15 is the results of the whole blood coagulation test (Imai-Nose method) of the powder-type hemostat prepared according to Example 1, Example 3 and Example 4.
  • Figure 19 is the result of the cytotoxicity test through the MTT assay of the powder-type hemostat prepared according to Examples 3 and 4 (elution conditions: 1mg / ml).
  • Figure 20 is the result of the cytotoxicity test through the MTT assay of the powder-type hemostat prepared according to Examples 3 and 4 (elution condition: 5mg / ml).
  • Figure 21 is the result of the cytotoxicity test through the MTT assay of the powder-type hemostat prepared according to Examples 3 and 4 (elution conditions: 10mg / ml).
  • Figure 23 is the result of the hemostatic ability evaluation (bleeding amount) through in vivo animal testing of the powdered hemostat prepared according to Example 1, Example 3 and Example 4.
  • Example 24 is the result of calculating the hemostatic ability evaluation (bleeding amount per hour) of the powdered hemostat prepared according to Example 1, Example 3 and Example 4 through in vivo animal experiments.
  • 25a to 25f are H&E staining results of powdered hemostats prepared according to Examples 1, 3 and 4.
  • 26a to 26f are Carstair's staining results of powdered hemostats prepared according to Examples 1, 3 and 4.
  • the hemostatic agent refers to a drug used for the purpose of stopping bleeding for bleeding symptoms, preferably in the form of a powder, and can be used in any part of the body where bleeding occurs.
  • FIG. 1 is a schematic diagram showing the structure of the powdered hemostatic agent of the present invention. Hereinafter, with reference to Figure 1 will be described for the powder type hemostatic agent of the present invention.
  • the present invention is a first polymer; a second polymer; and a cross-linking agent, wherein the first polymer and the second polymer are biocompatible polymers, and the first polymer and the second polymer are cross-linked by the cross-linking agent.
  • the biocompatible polymer is chitosan, starch, cellulose, alginate, gelatin, hyaluronic acid, sodium hyaluronate, collagen, polyvinyl alcohol (PVA), alginic acid, pectin, carrageenan, chondroitin (sulfate), dextran (sulfate), polylysine (polylysine), carboxymethyl titin, fibrin, agarose, pullulan, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), hydroxypropylcellulose (HPC), hydroxyethylcellulose (HEC), hydroxypropyl Methylcellulose (HPMC), sodium carboxymethylcellulose, polyalcohol, gum arabic, cyclodextrin, dextrin, glucose, fructose, trehalose, glucose, maltose, lactose, lactulose, fructose, turanose, melitose, melegitose , dex
  • the first polymer may include chitosan
  • the second polymer may include starch
  • Chitosan can be obtained by deacetylating chitin extracted from crustaceans such as crabs and shrimps in a concentrated alkaline solution.
  • chitosan As a biomedical material, chitosan has characteristics such as biocompatibility, biodegradability, non-toxicity, and non-immunity, and has self-sterilization properties capable of inhibiting the growth of bacteria and fungi.
  • chitosan as a cationic polymer, has mucoadhesive properties and can form clots during bleeding due to interaction with negatively charged platelets and red blood cells due to the presence of cations from aminoglucose.
  • Starch is a polymer composed of many glucose units linked by glycosidic bonds. It is the most common carbohydrate in the human diet and is present in large quantities in staple foods such as potatoes, maize, rice, wheat and cassava. Starch is generally composed of 20-25% amylose and 75-80% amylopectin, is inexpensive, and has excellent biocompatibility, non-toxicity, and biodegradability like chitosan. In addition, starch is used after being dissolved in water at 80 ° C. When dissolved in water, starch absorbs water and increases viscosity. An excellent hemostatic effect can be provided by increasing the viscosity of blood and increasing the concentration of blood cells.
  • the crosslinking agent is glyoxal, glutaraldehye, citric acid, At least one selected from the group consisting of N,N-methylene bisacrylamide, dialdehyde, oxalic acid and citric acid It may include, preferably may include glyoxal.
  • the crosslinking agent refers to a material that combines linear polymer monomer molecules through chemical bonds to form a polymer compound with a three-dimensional network structure.
  • biomolecular compounds can be cross-linked with each other through covalent bonds, hydrogen bonds, van der Waals bonds, and physical bonds. Accordingly, mucoadhesiveness and hemostatic properties of chitosan and viscosity are improved by starch, thereby providing performance as a hemostatic agent with excellent biocompatibility and blood absorption.
  • Glyoxal has the smallest dialdehyde group present and is a cross-linking agent that forms acetal bond or hemiacetal bond with hydroxyl group present in polysaccharide under acid catalyst, and forms primary amine and shiff base bond.
  • the powdered hemostatic agent is composed of particles including the first polymer and the second polymer, and the first polymer and the second polymer are crosslinked by the crosslinking agent.
  • the powder-type hemostatic agent may further include at least one selected from the group consisting of an adhesive, a calcium compound, and a hemostatic material.
  • the pressure-sensitive adhesive may include at least one selected from the group consisting of poly(vinyl pyrrolidone), PVP, pullulan, and dopamine, and preferably may include PVP. there is.
  • PVP is a non-toxic synthetic polymer because it is not absorbed through the gastrointestinal tract or mucous membrane, and as a powder soluble in water and polar solvents, it has the property of forming a film after the absorbed water evaporates.
  • PVP is an FDA-approved material and is widely used in daily life, such as biomaterials, cosmetics, and synthetic detergents.
  • Another characteristic of PVP is its excellent adhesive ability. This is because PVP has a strong attraction to other molecules due to its polarity.
  • the content of the adhesive may be 0.5 to 15 parts by weight, preferably 3 to 7 parts by weight, based on 100 parts by weight of the powdered hemostatic agent.
  • the pressure-sensitive adhesive may contain PVP, and the PVP solution may have a content of 0.5 to 15 mL compared to 100 mL of the PVP aqueous solution when preparing a powder-type hemostatic agent.
  • the calcium compound may include at least one selected from the group consisting of calcium chloride, calcium nitrate, calcium carbonate, calcium acetate, calcium carbonate, calcium sulfide, calcium carbonate, and calcium phosphate, preferably calcium chloride (CaCl 2 ). can do.
  • the hemostatic material may include at least one selected from the group consisting of thrombin, prothrombin, thromboplastin, fibrinogen, aprotinin, vitamin C and vitamin K, and preferably may include thrombin.
  • CaCl 2 activates prothrombin into thrombin
  • thrombin is a representative blood coagulation promoting factor that activates fibrinogen into fibrin.
  • Ca 2+ ions were introduced using thrombin and CaCl 2 , which are blood coagulation promoting drugs, to improve blood coagulation ability upon contact with blood.
  • Ca 2+ ions also exist in the body and activate prothrombin into thrombin, and thrombin activates fibrinogen into fibrin, which is involved in blood clot formation.
  • the size of the particles may be 1 to 500 ⁇ m, preferably 125 to 250 ⁇ m.
  • the adhesive, calcium compound and hemostatic material may be coated on the surface of the particle.
  • a first polymer solution and a second polymer solution are prepared, respectively, and mixed to prepare a first mixed solution (step a).
  • the weight ratio of the first polymer to the second polymer may be 1 to 7:7 to 1, preferably 1 to 3:3 to 1.
  • a crosslinking agent is added to the first mixed solution to prepare a crosslinked product in which the first polymer and the second polymer are crosslinked (step b).
  • the content of the crosslinking agent relative to 100 parts by volume of the first mixed solution may be 5 to 500 parts by volume, preferably 20 to 30 parts by volume.
  • step (b) the crosslinking reaction may be carried out for 2 to 72 hours, preferably for 10 to 15 hours.
  • the cross-linked product is lyophilized and pulverized to prepare a powdered hemostatic agent (step c).
  • step (b) mixing the crosslinked product with a solution containing at least one selected from the group consisting of an adhesive, a calcium compound and a hemostatic material to prepare a second mixed solution Step; may be further included.
  • the step (c) may be (c') preparing a powder-type hemostatic agent by lyophilizing and pulverizing the second mixed solution.
  • step (c) After freeze-drying in step (c), cooling under nitrogen gas conditions may be further included.
  • Chitosan solution was prepared by dissolving chitosan (low molecular weight 50,000 ⁇ 190,000 Da, Sigma-Aldrich Co.) in 100 mL of 2% acetic acid aqueous solution, heating 100 mL of distilled water to 80 ° C and dissolving starch (Daejung Co.) A starch solution was prepared.
  • a mixed solution was prepared by mixing the chitosan solution and the starch solution. The weight ratio of chitosan and starch in the mixed solution is 1.25:0.75 ( ⁇ 1.67:1).
  • a cross-linking reaction was performed for 12 hours by adding glyoxal (Sigma-Aldrich Co.) as a cross-linking agent to the mixed solution.
  • glyoxal Sigma-Aldrich Co.
  • the chitosan/starch aqueous solution was precipitated in 500 mL of acetone and stirred for 30 minutes in order to remove the unreacted cross-linking agent and obtain a cross-linked product in which chitosan and starch were cross-linked.
  • the precipitate and acetone were separated from the supernatant by centrifugation at 3000 rpm for 5 minutes, and the precipitate was finally separated through centrifugation after solvent exchange once in 500mL acetone and once in 500mL distilled water, a total of two times.
  • a swollen precipitate was obtained.
  • the obtained swollen precipitate was pre-frozen and then freeze-dried.
  • the freeze-dried sample was quenched using liquid nitrogen and ground in a mortar to prepare a chitosan/starch powder-type hemostatic agent.
  • Example 2 Preparation of chitosan/starch powder type hemostatic agent containing PVP
  • Example 1 the solvent exchange was completed by centrifugation, and PVP aqueous solution (molecular weight ⁇ 1,300,000 Da, Sigma-Aldrich Co.) was added to the centrifuged sample at a ratio of 1: 1 v / v%, and then a vortex mixer was used. and mixed. After preliminary freezing in a -80 ° C cryogenic freezer, freeze-drying was performed. The lyophilized sample was quenched using liquid nitrogen and then pulverized in a mortar to prepare a chitosan/starch powder-type hemostatic agent containing PVP.
  • Example 3 Preparation of chitosan/starch powder type hemostatic agent containing CaCl 2 /thrombin
  • the concentrations of the CaCl 2 solution and the thrombin solution were set to 20 mM and 100 ⁇ g/ml, respectively.
  • a CaCl 2 /thrombin solution was prepared by mixing the CaCl 2 solution and the thrombin solution.
  • Example 1 the solvent exchange was completed by centrifugation, and the CaCl 2 /thrombin solution was added to the centrifuged sample at a ratio of 1:1 v/v% and then mixed using a vortex mixer. After preliminary freezing in a -80 ° C cryogenic freezer, freeze-drying was performed. The lyophilized sample was quenched using liquid nitrogen and then pulverized in a mortar to prepare a chitosan/starch powder-type hemostatic agent containing CaCl 2 /thrombin.
  • Example 4 Preparation of chitosan/starch powder type hemostatic agent containing PVP/CaCl 2 /thrombin
  • the concentrations of the CaCl 2 solution and the thrombin solution were set to 20 mM and 100 ⁇ g/ml, respectively. After dissolving PVP in the CaCl 2 solution, a thrombin solution was added to prepare a PVP/CaCl 2 /thrombin solution.
  • Example 1 the solvent-exchanged sample was centrifuged, and the PVP/CaCl 2 /thrombin solution was added to the centrifuged sample at a ratio of 1:1 v/v% and mixed using a vortex mixer. After preliminary freezing in a -80 ° C cryogenic freezer, freeze-drying was performed. The lyophilized sample was quenched using liquid nitrogen and then pulverized in a mortar to prepare a chitosan/starch powder-type hemostatic agent containing PVP/CaCl 2 /thrombin.
  • the optimal content ratio was determined by considering the weight when the powder could no longer be absorbed by 0.1 g of the chitosan/starch powder-type hemostatic agent according to Example 1 as the maximum absorption rate. Absorption rate was calculated by Equation 1 below, and the results are shown in FIG. 2.
  • Equation 1 W 0 is the initial weight of the powder, and W max is the weight of the powder when it is absorbed to the maximum.
  • Example 1 the blood absorption rate according to the crosslinking time was observed in order to set an appropriate reaction time to provide sufficient time for the crosslinking reaction to occur as the crosslinking time was longer, and the results are shown in FIG. 3.
  • the crosslinking time is 2 hours in (a), 4 hours in (b), 12 hours in (c), 24 hours in (d), and 72 hours in (e).
  • the absorption rate is low because sufficient crosslinking is not performed in the initial 2 hours after the crosslinking agent glyoxal is added, and the network structure is not properly formed. As the reaction proceeds, the network structure is well formed. At 12 hours, the maximum absorption rate of 806% could be observed. After that, the tendency for the absorption rate to decrease as the reaction continues seems to decrease as the crosslinking degree increases and the network spacing of the polymer narrows. Through this, 12 hours with the best absorption rate was set as the optimal crosslinking time.
  • the absorption rate was confirmed to set the crosslinking agent content, and the results are shown in FIG. 4.
  • the content of the crosslinking agent is 1000 ⁇ l in (a), 500 ⁇ l in (b), 100 ⁇ l in (c), 50 ⁇ l in (d), 25 ⁇ l in (e), and 10 ⁇ l in (f).
  • the content of the cross-linking agent was 50 ⁇ l
  • the highest absorption rate was 1202%.
  • the higher the crosslinking agent content the lower the absorption rate due to the increase in the degree of crosslinking.
  • the amount of crosslinking agent is less than that of the polymer functional group, so the absorption rate is reduced because the crosslinking is not sufficiently performed.
  • the powder produced through pulverization after freeze-drying has a difference in blood absorption rate according to the particle size due to the nature of the powder.
  • the pulverized powder was sieved and classified into particle sizes of 125 ⁇ m or less, 125-250 ⁇ m, and 250-500 ⁇ m, and the blood absorption rate of the powder according to the particle size was measured to determine the optimal particle size.
  • the results are shown in FIG. .
  • the size of the particles is 250-500 ⁇ m in (a), 125-250 ⁇ m in (b), and 0-125 ⁇ m in (c).
  • the absorption rate was the highest at 1040.8% when the particle size was 125-250 ⁇ m.
  • Absorption rates at 250-500 ⁇ m and 0-125 ⁇ m were 866.2% and 696.6%, respectively. Based on the absorption rate, the particle size of 125-250 ⁇ m was selected as the optimal particle size.
  • FIG. 6 In order to confirm cross-linking of the chitosan/starch powder-type hemostatic agent prepared in Example 1, functional groups were analyzed using FTIR transmission, and the results are shown in FIG. 6 .
  • (A) is chitosan
  • (B) is starch
  • (C) is an FTIR spectrum of chitosan/starch powdered hemostatic agent prepared according to Example 1.
  • the adhesiveness was measured using TA instruments' DHR- ⁇ . Geometry (40 mm parallel plate, Peltier plate Steel, 110258), stage temperature (37°C), angular frequency (10 rad/s), and gap (3000 ⁇ 500 ⁇ m) were set as measurement conditions, and the adhesion in the actual tissue was observed. To this end, the experiment was conducted by attaching porcine skin (250-300 mm) to the geometry using an instant adhesive.
  • Example 7 is a graph comparing the adhesiveness of a chitosan/starch powder type hemostat prepared in Example 1 (Freeze dry), Pure PVP, and a chitosan/starch powder type hemostat containing 15wt% PVP.
  • FIG. 8 is a graph comparing the adhesiveness according to the PVP concentration of chitosan/starch powder-type hemostatic agents containing PVP prepared in Example 2. Referring to FIG. 8, it was confirmed that the adhesiveness of the powder improved as the PVP concentration increased.
  • the powder without PVP applied had the lowest adhesive force at -0.28 N, and the adhesive force of 15 wt% with the highest PVP concentration was -2.38 N, which was 881% higher than that of the powder without PVP applied.
  • FIG. 9 is a graph showing the blood absorption rate according to the PVP concentration of the chitosan/starch powder-type hemostatic agent containing PVP prepared according to Example 2.
  • the initial introduction of PVP hindered absorption rather than formation of a PVP film by the introduction of water and blood In the case of 3.0-7.0 wt%, it was confirmed that the ability to support water increased as the viscosity of the powder that absorbed water increased along with the formation of the PVP film. After 10.0 wt%, it was confirmed that the absorption rate decreased because contact between the powder and water/blood was blocked due to excessive PVP film formation.
  • a stainless steel tube rack to support the watch glass was placed, and water was filled in a constant temperature water bath to the extent that the bottom of the watch glass was submerged, and maintained at 37 ° C. Thereafter, the watch glass was put on the rack for 10 minutes to adjust the temperature, and then the experiment was conducted.
  • 300 ⁇ l of the CaCl 2 solution was placed on a watch glass, and 300 ⁇ l of PPP was introduced and mixed thereon, and the fibrin formation time was measured.
  • the tip of a 200 ⁇ l pipette tip was bent and the PPP into which the coagulation promoting factor was introduced was lifted and confirmed.
  • the CaCl 2 concentration with the shortest fibrin formation time was optimally set, and based on this, the optimal conditions for thrombin were set.
  • Fibrin formation time according to CaCl 2 concentration is shown in FIG. 10 .
  • the fibrin formation time was the shortest at 239 seconds when the concentration of CaCl 2 was 20 mM.
  • the concentration of Ca 2+ ions is low, fibrin formation is not properly involved, and when the concentration of Ca 2+ ions is high, activation of fibrinogen to fibrin is delayed, resulting in a V-shape.
  • 11 is a graph showing fibrin formation time according to thrombin concentration. Referring to FIG. 11, the higher the concentration of thrombin, the shorter the fibrin formation time, but at a concentration higher than 100 ⁇ g/ml, the fibrin formation time was similar to 0.13 seconds.
  • a scanning electron microscope (SEM, JSM-6380. Japen) was used to observe the shape and microstructure of the powdered hemostatic agent.
  • a scanning electron microscope is a type of electron microscope that scans and images the surface of a sample through an electron beam. When high-speed electrons are fired, the electrons collide with the sample surface and interact to analyze the ejection of materials such as electrons from the sample. to be.
  • the samples were coated with platinum to impart conductivity before observation using a sputter coater (108 auto, degree of vacuum: less than 10 Pa, Cressington Scientific Instruments Inc. UK). Accurate SEM images could be observed through the coating and were observed under conditions of accelerating voltage (10 kV) and working distance (15 mm).
  • FIGs 12a1 to 12i2 are a chitosan/starch powder type hemostat prepared according to Example 1
  • (B) is a chitosan/starch powder type hemostat containing CaCl 2 /thrombin prepared according to Example 3
  • (C) to (I) are SEM images of chitosan/starch powder-type hemostat containing PVP/CaCl 2 /thrombin prepared according to Example 4, and
  • (C) to (I) are PVP concentrations of 0.5 and 1, respectively. , 3, 5, 7, 10, 15 wt%.
  • Example 3 Surface of the chitosan/starch powder-type hemostat containing CaCl 2 /thrombin prepared in Example 3 using energy-dispersive X-ray spectroscopy (EDS) to identify CaCl 2 and thrombin introduced to promote blood coagulation. Elemental analysis was performed. Through elemental analysis, Ca 2+ of CaCl 2 and S in cysteine and methionine among the amino acid structures of thrombin were confirmed to confirm the presence of blood coagulation promoting factors in the powder phase and the degree of dispersion.
  • EDS energy-dispersive X-ray spectroscopy
  • (A) is a chitosan/starch powder type hemostat prepared according to Example 1
  • (B) is a chitosan/starch powder type hemostat containing CaCl 2 /thrombin prepared according to Example 3.
  • EDS analysis Referring to FIGS. 13a and 13b, it was confirmed that the Ca 2+ ions of the coated CaCl 2 were contained on the surface by about 6.48%. It is expected that element S was not detected due to the small effect of the ratio of cysteine to methionine present in thrombin. As a result of EDS mapping of Ca 2+ ions in yellow, it was confirmed that they were uniformly dispersed in all particles.
  • the Lee-White method was performed to measure the blood coagulation time of powdered hemostatic agents.
  • the Lee-White method is to put whole blood into a glass vial and set the time when the blood loses fluidity and clots as the blood coagulation time.
  • blood itself without introduction of the powdered hemostatic agent was set, and the powdered hemostatic agent prepared according to Example 1, Example 3 and Example 4 was set as the experimental group.
  • 10 mg of powdered hemostat in the vial 1 ml of whole blood was added and the time at which blood no longer flowed was recorded by tilting, and the results are shown in FIG. 14 .
  • the condition containing the blood coagulation promoting factor CaCl 2 and thrombin was about 3.5 times faster than the condition without the blood coagulation promoting factor.
  • the fastest time was 36 seconds at 7 wt% of PVP, but there was no significant difference when compared to other PVP conditions.
  • the blood coagulation ability of the powder was improved due to the introduction of the blood coagulation promoting factor, and it was confirmed that the blood coagulation ability was not inhibited by the introduction of PVP.
  • the Imai-Nose method was performed to confirm the clot formation ability of the powdered hemostatic agent.
  • a stainless steel centrifugal tube tube rack was placed in a constant temperature water bath set at 37 ° C., and a watch glass was placed on it, and the temperature was maintained for 10 minutes. Thereafter, 10 mg of a powder-type hemostatic agent was introduced and 200 ⁇ l of blood was added and the experiment was performed while mixing. As a control, the thrombogenicity was observed without applying the powder.
  • the reaction times of blood and powder were observed at 1, 3, 5, 7, 10, and 20 minutes, respectively, and when the reaction time was reached, 5 ml of distilled water was added to terminate the blood coagulation reaction.
  • the resulting clot was separated, fixed with 35% formaldehyde for 10 minutes, soaked in distilled water, washed for 5 minutes, and dried for 24 hours. Then, the weight of the thrombus was measured according to the reaction time, and the results are shown in FIG. 15 .
  • the degree of thrombosis was about 7-8 times higher than that of the control group. It was observed that hemostatic agents formed clots better. It seems that the cross-linked chitsoan/starch powder helped blood coagulation by absorbing blood and reducing the amount of remaining blood. In addition, as a result of observing thrombus formation according to the PVP concentration, it was confirmed that the introduction of PVP did not inhibit the formation of thrombi.
  • the red blood cell hemolysis test is a test method for quantifying the remaining red blood cells according to the blood coagulation reaction. As the red blood cells are hemolyzed, the hemoglobin in the red blood cells is released, and the blood coagulation ability is confirmed by measuring the absorbance at this time.
  • a stainless steel centrifugal tube tube rack was placed in a constant temperature water bath and a watch glass was placed thereon. After introducing 5 mg of powdered hemostatic agent on the watch glass, water was filled to submerge the bottom of the watch glass, and the temperature was maintained at 37° C. for 10 minutes. Thereafter, 200 ⁇ l of blood was introduced and carefully mixed to initiate the coagulation reaction. As a control, 200 ⁇ l of only blood not introduced with powder was placed in a watch glass. The reaction time was set to 1, 3, 5, 7, 10, and 20 minutes. When the reaction time was over, 5 ml of distilled water was added to terminate the blood coagulation reaction, and blood clots and powder were removed, followed by hemolysis of red blood cells for 5 minutes.
  • the absorbance was about 18% lower than that when the factor was not introduced. This indicates that a blood clot is formed by a blood coagulation reaction, and there are few remaining red blood cells, and many blood clots are generated. As there was no significant difference in the absorbance by the remaining red blood cells according to the PVP concentration, it was confirmed that PVP did not affect clot formation.
  • Plasma protein coagulation occurs by adding calcium ions to plasma decalcified by anticoagulants. Using this, a plasma recalcification time (PRT) measurement method was performed to investigate the interaction between the powdered hemostatic agent and plasma proteins.
  • PRT plasma recalcification time
  • PRT measurement measures the time until fibrin is formed after adding calcium ions to PPP obtained by centrifuging whole blood.
  • a powdered hemostatic agent prepared according to Example 4 was used as an experimental group. PPP was obtained by centrifugation of whole blood containing anticoagulant, and the separated PPP was stored on ice and used within several hours. In addition, since an anticoagulant was included, a 0.0125 M CaCl 2 solution was added to suppress the anticoagulant.
  • a stainless steel tube rack was placed in a constant temperature water bath, a watch glass was placed on it, the water was adjusted so that the bottom of the watch glass could be submerged, and a temperature of 37 ° C was applied. Thereafter, 20 mg of a powdered hemostatic agent was placed on a watch glass while adjusting the temperature with a constant temperature water bath for 10 minutes, and 300 ⁇ l of PPP and 300 ⁇ l of a 0.025 M CaCl 2 solution were introduced to observe the fibrin formation time. Upon observation, fibrin formation was observed while lifting the plasma by bending the end of a 200 ⁇ l pipette tip, and the results are shown in FIG. 18 .
  • the fibrin formation time was about 4 to 5 times faster when the blood coagulation accelerator was included than when it was not included.
  • the condition in which PVP was introduced took the longest fibrin formation time among the conditions in which PVP was introduced, at 54.66 seconds at 1.0 wt%, but there was no significant difference from other concentration conditions, so it was confirmed that PVP introduction did not inhibit fibrin formation.
  • 1 ⁇ 10 4 cells of NIH/3T3 cells were dispensed into wells of a 96-well cell culture plate and cultured for 24 hours in an incubator at 37°C and 5% CO 2 environment.
  • the elution medium was eluted for 24 hours by applying 1, 5, and 10 mg of powdered hemostat per 1 ml of the medium.
  • the medium of the 96-well cell culture plate was suctioned, and 100 ⁇ l of the dissolution medium was dispensed into each well and cultured for another 24 hours.
  • MTT solution made of DMEM medium (9): MTT stock solution (1) was added and reacted for 1-4 hours. After 1-4 hours, after removing the MTT reagent, 100 ⁇ l of DMSO was added to dissolve insoluble formazan, and the absorbance was measured at 540 nm using an ELISA microplate reader. 20 (elution conditions: 5 mg/ml) and FIG. 21 (elution conditions: 10 mg/ml).
  • avertin stock solution by mixing 25 g avertin (2,2,2-tirbromoethanol) and 15.5 ml tert-amyl alcohol (2-methyl-2-butanol) to be used as an anesthetic for 12 hours in a dark room.
  • the amount of blood loss is calculated by measuring the weight of the filter paper, powder, and disposable gauze that have absorbed blood.
  • a control group was set in a state in which nothing was introduced, and ARISTA-AH, a commercially available highly absorbable hemostatic agent, was used as a positive control group.
  • ARISTA-AH a commercially available highly absorbable hemostatic agent
  • the powdered hemostatic agent prepared according to Example 1, Example 3 and Example 4 was set as an experimental group.
  • a powder-type hemostat containing 3.0 wt% PVP and 10.0 wt% PVP was used.
  • the results for bleeding time are shown in FIG. 22 .
  • the bleeding time of the control group was about 5 minutes and 31 seconds
  • the positive control group, ARISTA-AH was about 2 minutes and 55 seconds.
  • the speed of hemostasis was 2-3 times faster than that of conditions without it.
  • chitosan/starch powder type hemostat containing 3 wt% PVP/CaCl 2 /thrombin was the fastest showed speed. It seems that the powder was not lost along with bleeding during the hemostasis process and was well adhered to the tissue, so that the bleeding was stopped in a short time.
  • the results of blood loss are shown in FIG. 23 .
  • the control group and ARISTA-AH were 3439.8 mg and 1533.6 mg, respectively, the chitosan/starch powder type hemostat in the experimental group was 1202.8 mg, and the chitosan/starch powder type hemostat containing CaCl 2 /thrombin was 606.2 mg, 3wt%.
  • Chitosan/starch powder-type hemostat containing PVP/CaCl 2 /thrombin and chitosan/starch powder-type hemostat containing 10wt% PVP/CaCl 2 /thrombin obtained 273 mg and 974.2 mg, respectively.
  • the control group and ARISTA-AH are 637.5 mg/min and 459.64 mg/min
  • the experimental group is chitosan/starch powder type hemostat, chitosan/starch powder type hemostat containing CaCl 2 /thrombin, 3 wt% PVP/ Chitosan/starch powder-type hemostat containing CaCl 2 /thrombin and chitosan/starch powder-type hemostat containing 10wt% PVP/CaCl 2 /thrombin were 449.91 mg/min, 291.81 mg/min, 157.13 mg/min, and 404.15 mg, respectively. The value of /min was shown.
  • blood coagulation accelerators help to form clots, they can prevent re - bleeding, but they do not form hard fibrin, so they are easily removed along with the powder.
  • hemostatic agents effective hemostasis was possible based on appropriate adhesiveness and high absorption rate, so re-bleeding did not occur.
  • chitosan/starch powder type hemostat containing 10wt% PVP/CaCl 2 /thrombin with excessive adhesiveness is removed together with the blood clot generated due to tissue and strong adhesion in the process of removal after spraying saline, so it can be re-removed. It seems that bleeding occurs and the hemostatic effect is reduced. Through this, it was confirmed that the chitosan/starch powder type hemostatic agent containing 3wt% PVP/CaCl 2 /thrombin had better hemostatic ability than commercially available ARISTA-AH.
  • hematoxylin, a basic dye, and eosin, an acidic dye are sequentially stained to stain cell nuclei in blue, and cytoplasm, cell membranes, red blood cells, and collagen are stained in red.
  • the collected tissue is first put into a 10% formalin solution to fix it, and then washed with a xylene solution. Thereafter, the tissue was put in paraffin, solidified, and then thinly sectioned with a microtome to a thickness of 5 ⁇ m, washed twice in xylene for 10 minutes for deparaffinization, and then dipped in alcohol for 5 minutes to remove xylene. After alcohol washing, proceed with hematoxylin staining in hematoxylin solution for 8 minutes, and then wash in running water for 5 minutes.
  • (A) is a control group
  • (B) is ARISTA-AH
  • (C) is a chitosan/starch powder type hemostat
  • (D) is a chitosan/starch powder type hemostat containing CaCl 2 /thrombin
  • (E) is a chitosan/starch powder-type hemostat containing 3wt% PVP/CaCl 2 /thrombin
  • (F) is an H&E staining result of a chitosan/starch powder-type hemostat containing 10wt% PVP/CaCl 2 /thrombin. Referring to FIGS. 25A to 25F , it was confirmed that the wound area was stained red due to red blood cell aggregation.
  • A is a control group
  • B is ARISTA-AH
  • C is a chitosan/starch powder type hemostatic agent
  • D is CaCl 2 /thrombin-containing chitosan/starch powder type hemostat
  • E is chitosan/starch powder type hemostatic agent containing 3wt% PVP/CaCl 2 /thrombin
  • F is chitosan containing 10wt% PVP/CaCl 2 /thrombin /starch This is the result of Carstair's staining of a powder-type hemostatic agent. Referring to FIGS. 26A to 26F , fibrin formed during hemostasis was confirmed through orange staining at the wound site under all conditions.
  • chitosan/starch powder type hemostat containing CaCl 2 /thrombin applied with CaCl 2 and thrombin chitosan/starch powder type hemostat containing 3wt% PVP/CaCl 2 /thrombin, than control group and ARISTA-AH
  • chitosan/starch powder type hemostatic agent containing 10wt% PVP/CaCl 2 /thrombin was applied, clearer red blood cell aggregation was observed at the bleeding site. This seems to be the result of platelet aggregation of Ca 2+ ions and rapid hemostasis of thrombin.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Materials Engineering (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Immunology (AREA)
  • Surgery (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Hematology (AREA)
  • Zoology (AREA)
  • Diabetes (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Materials For Medical Uses (AREA)

Abstract

La présente invention concerne un agent hémostatique de type poudre comprenant un polymère biocompatible et son procédé de préparation. Plus particulièrement, la présente invention possède une excellente biocompatibilité grâce à l'utilisation d'un polymère naturel et présente une vitesse d'absorption sanguine élevée à travers une structure réticulée. De plus, la présente invention comprend une substance hémostatique de telle sorte que la vitesse de thrombogenèse est rapide, permet la formation de nombreux thrombus de telle sorte que la coagulabilité du sang est excellente, comprend un adhésif de telle sorte qu'une propriété d'adhérence tissulaire peut être augmentée, et offre d'excellents effets hémostatiques.
PCT/KR2022/009675 2021-07-05 2022-07-05 Agent hémostatique de type poudre comprenant un polymère biocompatible et son procédé de préparation WO2023282583A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2021-0087909 2021-07-05
KR1020210087909A KR20230008274A (ko) 2021-07-05 2021-07-05 생체적합성 고분자를 포함하는 파우더형 지혈제 및 그의 제조방법

Publications (1)

Publication Number Publication Date
WO2023282583A1 true WO2023282583A1 (fr) 2023-01-12

Family

ID=84800710

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2022/009675 WO2023282583A1 (fr) 2021-07-05 2022-07-05 Agent hémostatique de type poudre comprenant un polymère biocompatible et son procédé de préparation

Country Status (2)

Country Link
KR (1) KR20230008274A (fr)
WO (1) WO2023282583A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116059437A (zh) * 2023-02-27 2023-05-05 南通大学 一种抗收缩血液接触组织粘合材料的制备方法及应用
CN116421771A (zh) * 2023-04-10 2023-07-14 东莞博捷生物科技有限公司 一种新型多孔淀粉止血粉及其制备方法和应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115970048B (zh) * 2023-02-24 2024-06-11 海孵(海南自贸区)医疗科技有限责任公司 一种高保湿性的聚乙二醇液体敷料及其制备方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100885178B1 (ko) * 2008-03-19 2009-02-23 고려대학교 산학협력단 점착성 약제 조성물
KR20170007024A (ko) * 2015-07-10 2017-01-18 (주)헵틸와이 이중 가교구조의 생체친화성 지혈제 및 그 제조방법
KR20210052099A (ko) * 2019-10-31 2021-05-10 주식회사 삼양바이오팜 분말형 지혈제 조성물 및 이의 제조 방법
KR20220072149A (ko) * 2020-11-25 2022-06-02 금오공과대학교 산학협력단 파우더형 지혈제 제조방법 및 그에 따른 파우더형 지혈제

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100885178B1 (ko) * 2008-03-19 2009-02-23 고려대학교 산학협력단 점착성 약제 조성물
KR20170007024A (ko) * 2015-07-10 2017-01-18 (주)헵틸와이 이중 가교구조의 생체친화성 지혈제 및 그 제조방법
KR20210052099A (ko) * 2019-10-31 2021-05-10 주식회사 삼양바이오팜 분말형 지혈제 조성물 및 이의 제조 방법
KR20220072149A (ko) * 2020-11-25 2022-06-02 금오공과대학교 산학협력단 파우더형 지혈제 제조방법 및 그에 따른 파우더형 지혈제

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
KIM DONGJIN: "Fabrication and Characterization of Absorbable Powder-type Hemostatics Containing Blood Coagulation Factors", MASTER'S THESIS, DEPARTMENT OF POLYMER SCIENCE ＆ ENGINEERING, GRADUATE SCHOOL KUMOH NATIONAL INSTITUTE OF TECHNOLOGY, 1 January 2019 (2019-01-01), XP093022425 *
SUKNUNTHA KRIT, TANTISHAIYAKUL VIMON, WORAKUL NIMIT, TAWEEPREDA WIRACH: "Characterization of muco- and bioadhesive properties of chitosan, PVP, and chitosan/PVP blends and release of amoxicillin from alginate beads coated with chitosan/PVP", JOURNAL DRUG DEVELOPMENT AND INDUSTRIAL PHARMACY, NEW YORK, NY, US, vol. 37, no. 4, 1 April 2011 (2011-04-01), US , pages 408 - 418, XP055846962, ISSN: 0363-9045, DOI: 10.3109/03639045.2010.518149 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116059437A (zh) * 2023-02-27 2023-05-05 南通大学 一种抗收缩血液接触组织粘合材料的制备方法及应用
CN116059437B (zh) * 2023-02-27 2023-12-19 南通大学 一种抗收缩血液接触组织粘合材料的制备方法及应用
CN116421771A (zh) * 2023-04-10 2023-07-14 东莞博捷生物科技有限公司 一种新型多孔淀粉止血粉及其制备方法和应用
CN116421771B (zh) * 2023-04-10 2024-05-07 东莞博捷生物科技有限公司 一种新型多孔淀粉止血粉及其制备方法和应用

Also Published As

Publication number Publication date
KR20230008274A (ko) 2023-01-16

Similar Documents

Publication Publication Date Title
WO2023282583A1 (fr) Agent hémostatique de type poudre comprenant un polymère biocompatible et son procédé de préparation
Sanandiya et al. Tunichrome-inspired pyrogallol functionalized chitosan for tissue adhesion and hemostasis
RU2704259C2 (ru) Пористый гемостатический продукт со свойством адгезии к тканям
KR101401873B1 (ko) 속효성 무수 밀봉제 및 그의 사용 및 제조 방법
Li et al. Injectable polysaccharide hybrid hydrogels as scaffolds for burn wound healing
WO2016200218A1 (fr) Structure poreuse et son procédé de fabrication
US6923961B2 (en) Chemically activated carboxypolysaccharides and methods for use to inhibit adhesion formation and promote hemostasis
JP2023118919A (ja) 止血材
Wei et al. Self-assembling RATEA16 peptide nanofiber designed for rapid hemostasis
WO2016010330A1 (fr) Compostion de mousse polymère, procédé de préparation de la composition de mousse polymère à l'aide de celle-ci, et mousse polymère pour emballage
RU2669569C2 (ru) Формованный пластинчатый продукт и гемостатический материал
WO2013060769A2 (fr) Compositions hémostatiques
WO2018106076A1 (fr) Matériau styptique comprenant une poudre de carboxyméthylcellulose de calcium et une poudre de composé de chitosane soluble dans l'eau, et son procédé de production
Yan et al. A keratin/chitosan sponge with excellent hemostatic performance for uncontrolled bleeding
Teng et al. A glycosylated and catechol-crosslinked ε-polylysine hydrogel: Simple preparation and excellent wound hemostasis and healing properties
Feng et al. A hemostatic keratin/alginate hydrogel scaffold with methylene blue mediated antimicrobial photodynamic therapy
KR20200051644A (ko) 슬관절 치환술을 위한 트라넥삼산 스프레이
Wang et al. A contact-polymerizable hemostatic powder for rapid hemostasis
JP5914465B2 (ja) 止血剤組成物
Chu et al. Physico-mechanical and biological evaluation of an injectable m-TG cross-linked thrombin loaded amended gelatin hemostat to heal liver trauma
CN112300418A (zh) 一种可粘附高效止血微球及其制备方法
CN112999408B (zh) 冻凝胶支架的制备方法及应用
AU2016261108B2 (en) Haemostatic compositions
Liu et al. Injectable plant-derived polysaccharide hydrogels with intrinsic antioxidant bioactivity accelerate wound healing by promoting epithelialization and angiogenesis
Jiang et al. An adenine-modified chitosan cross-linked 8arm-PEG-CHO hydrogel for promoting infected wound healing

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 22837935

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE