Hemostatic Agent

Abstract

The invention relates to the fields of materials science and medicine, particularly a novel hyaluronan-based hemostatic agent suitable for use in medicine for stopping bleeding and promoting wound healing. The proposed hemostatic agent stops bleeding very rapidly and exhibits good adhesion to body tissues. This technical result is achieved because the hemostatic agent is a solid compound of thrombin and hyaluronan, with a thrombin content of not less than 0.01 IU / cm² and not more than 10⁹ IU / cm². The hemostatic agent may also contain an antiseptic component and / or an analgesic component and / or a component that promotes wound healing.

Description

TARIFF Hemostatic Agent The invention relates to the field of human needs, more specifically hemostasis. medical devices that can be used in the provision and healing of wounds and 5 It is related to the compositions. The hemostatic agent, preferably human thrombin. hyaluronan (hyaluronic acid) together with thrombin Biocompatible thanks to the use of acid salts and solvates. It is bioresorbable. Hyaluronic acid, D-glucuronic acid and (N)-acetyl-D-glucosamine again It is a polysaccharide composed of repeating units. Depending on the number, hyaluronic acid is low molecular weight (100,000– up to 300,000 Da) or high molecular weight (from 500,000 Da) (starting from and in some cases exceeding 1,000,000 Da) are classified as such. 15 Hyaluronic acid forms salts and solvates; these are also called solvates of salts. This includes the term "hyaluronan," which describes this broader class of compounds. While commonly used for hyaluronic acid, the term "hyaluronate" refers to a different type of hyaluronic acid. It refers to a subgroup consisting of various salts. Hyaluronic acid, ammonium, magnesium and alkali metal salts, and some They share common characteristics, including, in particular, solubility in water. (Petr Snetkov, Kseniia Zakharova, Svetlana Morozkina, Roman Olekhnovich, Mayya Uspenskaya. Hyaluronic Acid: The Influence of Molecular Weight on Structural, Physical, Physico-Chemical, and 25 Degradable Properties of Biopolymer, p. 3. doi: 10.3390 / polym12081800). - 1 - - 2 - In the current state of the art, when a solution of a substance is dried, Trace amounts of solvent often remain in the resulting solid phase ("residue"). moisture). Such substances are commonly referred to as solvates. In addition, some substances, including hyaluronan, are highly hygroscopic. 5 It can even absorb moisture from the air. According to scientific and technical publications, in the bodies of mammals, including humans Hyaluronic acid is mainly in the form of its high molecular weight sodium salt. It is found. Hyaluronan undergoes continuous resorption in living organisms. 10 is undergoing and being synthesized; the total amount found in the human body Up to one-third of hyaluronan is reabsorbed and replenished each day. It is being synthesized. Scientific and technical publications also state that hyaluronan chains in solutions are 15 various secondary and tertiary structures, including water retention network structures It is reported that it can create. On an industrial scale, hyaluronans are mainly from biological sources. (mainly from rooster combs) by extraction or biotechnological 20 It is produced by synthesis. Industrially produced hyaluronan has a volume of 0.05 to 100%. Fine, amorphous particles with sizes ranging between 0.25 mm. It is a white powder. - 3 - Thrombin is a known protein component of human blood (Earl W. Davie, John D. Kulman, An Overview of the Structure and Function of Thrombin. doi: 10.1055 / s-2006-939550). Thrombin on the activation of blood clotting, It converts fibrinogen into fibrin and activates other clotting factors. Fibrin forms a network structure that traps red blood cells; red blood cells, 5 by forming a clot with platelets, thus achieving hemostasis. This leads to its provision. Including chitosan, cellulose, hyaluronic acid and their derivatives. Numerous polysaccharide-based hemostatic agents are known. With these 10 together, it plays a role in the formation of the extracellular matrix in vertebrates. due to the highest biocompatibility of hyaluronan with humans It should be taken into consideration that it exhibits; in contrast, cellulose and chitosan respectively. similar functions in plants and crustaceans, fungi and insects It brings. Cellulose and chitosan occur naturally in the human body 15 It is not available. Specifically, a biomaterial (a porous matrix) and a matrix of a a hemostatic sponge made of a material that increases adhesion to the organ It is known (US 8,771,258). Porous matrix biomaterials, primarily from the 20 It consists of various substances, including polysaccharides, primarily chitosan. The sponge also contains two crosslinkers, primarily polyethylene glycols. It also contains an adhesive material with components that are foreign to the human body. In addition to containing these substances, another characteristic of the sponge described above is... The disadvantage is that hemostasis takes approximately 2 minutes to occur, as shown in Example 10. It is a process that takes place relatively slowly. - 4 - Additionally, it is a bioadhesive material that does not contain oxidized cellulose or gelatin. A hemostatic sponge containing this composition is also known (US Patent). (2021 / 0228764). The presence of cellulose, which is foreign to the human body, in this This is a disadvantage of the material. 5 With a fast-acting, dry sealant, the production of the material in question and application methods are known (WO 2008 / 016983). This composition, forming a porous material by cross-linking under suitable conditions 10 from two cross-linkable components and a hydrogel-forming component It consists of cross-linking components such as polyethylene glycol. It is used. This substance is used with thrombin in various concentrations. It was used in combination (Example 23); however, thrombin was used as an additional component. No benefit has been observed. Among the disadvantages of this method is hemostasis. the time required is relatively long (from 1 minute in Example 22 to Example 15) (up to 2 minutes at 26) and a substance foreign to the human body The use of polyethylene glycol is involved. It is known to be a hemostatic, antiseptic and wound healing sponge (RU 2). 226,406 patents). The sponge is made from denatured collagen (gelatin) and has its own 20 It can absorb up to 45–50 times its weight in blood. This substance Among the disadvantages is that it can swell, affecting nerves and blood. The risk of compression and associated adverse effects when used near blood vessels. Due to the consequences, the hemostasis period is prolonged (2–8 (minutes) is included. 25 - 5 - Ionic polysaccharide-based for use as wound and burn dressings. A porous plastic material is known (patent RU 2 762 729). Porous plastic material; alginates, pectins, carrageenans, chondroitin They are produced from sulfates or chitosan and its derivatives. In this material Hyaluronan is not used as the matrix material. This material has 5 Its hemostatic effect is unknown. In particular, according to Claim 2, the aforementioned The material is rendered insoluble in water. A micro and nanostructured bioplastic material is known (RU 2). 481,127 patents). The material in question consists of 10 components of hyaluronic acid. Proteoglycans are based on a nanostructured matrix. It contains glycoproteins, fibrillar proteins, and an antiseptic substance. The material is used in the treatment of burns and has a hemostatic effect. It is unknown. A method for producing modified hyaluronic acid. It is known (patent RU 2 191 782). The resulting chemically modified The film, when applied, supports active tissue regeneration and reduces inflammation. It prevents adhesions but reduces biodegradability. This The hemostatic effect of the material is unknown. 20 Hyaluronic acid derivatives, especially benzyl ester of hyaluronic acid, a biomaterial containing and aimed at preventing postoperative adhesions It is known (patent RU 2 177 332) that this material has a hemostatic effect. It is unknown. 25 - 6 - A wound dressing with hemostatic properties is known (RU 2 624 242). (patented). The wound dressing is made of a bacterial cellulose-based biopolymeric material. Including up to 10% hemostatic agent and up to 3% antimicrobial agent. It consists of bioactive components, including a hemostatic agent. It shows a relatively rapid effect. The disadvantages of this method include 5. the use of cellulose, a material foreign to the human body and thrombin, a valuable component obtained from the patient, is relatively high. This includes consuming amounts (between 0.5 and 150 IU / cm²). A hemostatic compound containing hyaluronic acid is known (RU 2 486 921 10 (patented). Composition: gelatin with at least 10% natural (chemical cross-linking). hyaluronic acid or hyaluronic acid derivatives (not treated with binders) It contains. The composition requires dry heat at temperatures between 110°C and 200°C. It is stabilized by applying this method. The reduction in swelling indicates that this compound... While it has an advantage, one of its disadvantages is that hemostasis is relatively 15 This occurs more slowly, and it takes 2 minutes or more to achieve hemostasis. It takes longer. A carrier containing fibrinogen and thrombin is known (US 7,399,483). patent). The carrier mentioned above, in particular, has the following characteristics: 20 It is similar to the invention: (1) it is used for hemostasis purposes; (2) hyaluronic acid, together with some other substances, as a carrier matrix can be used (however, hyaluronic acid is used in specific application forms) The use of the acid is not specified, only lyophilized and foamed. collagen, gelatin, oxidized cellulose and polyglycolic acid / dioxanone 25 sponges were used); (3) carrier, thrombin (1.5–2.5 IU / cm²) and Coated with fibrinogen (4.3–6.7 mg / cm²). Disadvantages of this method. - 7 - between them, both are valuable components derived from human resources. the consumption of relatively high amounts of thrombin and fibrinogen Its use is included. Furthermore, in medical literature (VA Gorsky, A. M. Zryanin, MA Agapov. The Efficiency of TahoComb in Hepatobiliary Surgery / / Modern Technologies in Medicine. 2011, no. 2, p. 61–68), 5 Another disadvantage of the method is the requirement for achieving hemostasis. It is stated that the duration is relatively long and lasts approximately 3 minutes. Proteins, aliphatic polyesters, and various water-soluble polymers. a 10 containing compounds and some variants exhibiting hemostatic effects The shaped sheet product is known as (EP 2851095). The invention description states that the hemostatic effect is due to all of the materials in question. not shown by (in particular, as claimed in this invention) Combination of thrombin with a water-soluble polymer [hyaluronan] 15 (by) it is stated. Instead, the effect in question is due to fibrinogen-containing an aliphatic polyester containing thrombin with a water-soluble polymer layer overlapping layers (Example 21) or these layers beforehand certain laminated versions (Examples 24, 27, 29 and 31) This is due to combinations. Also in Example 30, thrombin (24,2 20 (at a concentration of IU / cm²) and for a composition containing an aliphatic polyester 3 A hemostatic effect is suggested to occur after a few minutes. However, a one that contains only a water-soluble polymer and thrombin any example demonstrating that the compound has a hemostatic effect 25 It is not provided. The source document also states (page 12 of the specification, first) - 8 - (paragraph), the structural design of the product shaped using the proposed method. It is claimed that its properties are superior to those obtained by lyophilization. is being pursued. In contrast, the claim in the present invention Hemostatic materials also exhibit superior structural properties. It is produced by lyophilization; this is stated by the applicant in 5 the suggested materials and the materials described in the referenced document This reveals that they are different. Therefore, the hemostatic material claimed in the present invention and EP 10 shaped sheet materials according to patent 2851095 combinations that differ significantly in terms of their composition It shows. The claims of patent EP 2851095 assert that neither the hemostatic effect nor thrombin It explicitly includes its presence. Furthermore, the specification states that thrombin (0.01 IU / cm² 15 (at concentrations of up to 100 IU / cm²) and a water-soluble one such as hyaluronan Any information regarding the hemostatic effect of a polymer-containing compound? It is not present. Specifically, in the first paragraph of page 7 of the specification, Thrombin content below 0.01 IU / cm² does not have any hemostatic effect. It does not provide, and thrombin content above 100 IU / cm² is 20 It is stated that this makes the shaped sheet product brittle. Furthermore, EP 2851095 document specifies thrombin levels in the range of 0.01 IU / cm² to 100 IU / cm². any hemostatic effect for a water-soluble polymer containing It does not explain. Hemostatic effect is only due to thrombin (24.2 IU / cm² For a combination of an aliphatic polyester with (at a concentration) 25 in Example 30. It is shown that a water-soluble polymer containing fibrinogen is combined with thrombin. In other examples where an aliphatic polyester containing the same material is present, the aliphatic - 9 - The thrombin content in polyester ranged from 14.7 IU / cm² (Sample 20) to 31.39 IU / cm² (Example 15) varies between these. Therefore, water containing thrombin in the range of 0.01 IU / cm² to 100 IU / cm² The hemostatic effect of a soluble polymer composition is one of the five known applications of the technique. This is unknown from the status (patent EP 2851095). Also, Dittrich, M. and Snejdrova, E. (2014), Cyclic Swelling as a Phenomenon Inherent to Biodegradable Polyesters, Journal of Pharmaceutical Sciences, 103(11), Publication 3560–3566, doi:10.1002 / jps.24146, biodegradable aliphatic This explains why polyesters tend to swell; this is stated in claim 10. This is a known disadvantage that is resolved by a technical solution. The invention described in patent EP 2851095 also includes a “shaped plate”. product (“laminating, weaving, knitting one or more fibers”) or created by being subjected to another process”) the subject of the claim 15 The necessity of producing a shaped product, production This complicates the process; this creates a disadvantage and For example, potential in applications requiring a free-flowing material. This limits its areas of use. In contrast, a powdered form Hemostatic material, application of a sheet-form product is difficult 20 in terms of achieving hemostasis in narrow or restricted-access areas This will be extremely useful. The requested material shaping It does not require and is available in a much wider variety of ready-to-use forms. They can be produced, thus expanding the technical range of existing hemostatic agents. It expands. 25 - 10 - Thrombin or fibrinogen with a polymeric carrier (e.g., hyaluronic acid) A pharmaceutical composition containing the following active ingredients is known; dispersed within or on the surface of a second (polymeric) carrier It is located inside a glassy carrier (WO 2013 / 004838 A1). According to the composition, it provides hemostasis in moderate bleeding. The time required to achieve this varies between 2 and 10 minutes (p. 24, paragraph 4). The presence of thrombin within a glassy carrier makes this compound one of the basic 10. It is one of its characteristics. The composition belonging to the known state of this technique... The disadvantage is that the time required to achieve hemostasis is relatively long. It must last, that is, 120 seconds or more. The technical problem of the invention is to expand the technical range of existing hemostatic agents by 15 The goal is to expand the area and shorten the time required to achieve hemostasis. The technical result of the invention is biocompatible, bioresorbable, and non-swelling. invisible, adheres to body tissues and is essential for maintaining hemostasis. A novel hemostatic based on hyaluronan and thrombin that reduces the required time. 20 It is the creation of matter. Using various forms of hyaluronan and thrombin concentrations Preliminary experiments unexpectedly revealed that the effect in question; hemostatic agent, natural hyaluronan 25, both of which are found in solid form. - 11 - and contains a compound containing thrombin, and the thrombin content is 0.01 obtained if it is not less than IU / cm² and not more than 109 IU / cm² This has been shown to be the case. In some forms of administration, hyaluronan is the most effective. It can exist at least as a partially crystallized polymer. The effect in question is that hyaluronan binds water in the plasma, causing plasma 5 increasing the concentration of its components and producing more effective and faster platelets This is achieved by promoting its aggregation, while the substance contains Thrombin converts natural fibrinogen into fibrin and hyaluronan networks. It completes the process by forming fibrin networks that immobilize red blood cells. The process involves the formation of a clot (thrombosis) that effectively stops bleeding. This leads to its formation. The reactivity of hyaluronan affects the substance. Thrombin concentration is crucial for thrombus formation and bleeding cessation. It is directly related to the speed. Hemostatic agent, additional properties. to improve or enhance the effectiveness of other blood clotting agents factors, cell growth factors, antiseptics, anesthetics and / or 15 wound-healing agents (such as methyluracil or dexpanthenol) It may include substances that do not contribute to its primary function. They may also contain excipients, riboflavin and silicon being examples. These can be given as; the excipients in question are distinguishable from the active and non-active surfaces. 20 It can be used. The substance, such as ethylene oxide or radiofrequency radiation, It can be sterilized using methods known to the expert. The proposed technical solution is novel and is not patented or published in scientific and technical journals. Not specified. 25 - 12 - The recommended hemostatic agent is used, for example, to achieve hemostasis and wound healing. It can be used in its improvement. The following examples illustrate the composition, properties, and characteristics of hemostatic agents. It explains the advantages. 5 Example 1 Produced by Swedlight AB (Sweden), it has a molecular weight of 11.6 × 10⁵. g sodium hyaluronate obtained by biotechnological method, 2 L distilled water dissolved in water and placed in a tray with a surface area of ​​1.478 cm² It was spilled. Later, the Hemophilia Patients Inter-Association Charity Renam Research and Production, Public Institution for Persons with Disabilities PG-9A thrombin, produced by the unit, for hemostasis tests. The reagent solution (with an activity of 9 IU) was added to the hyaluronate solution. and is evenly distributed on the surface. The resulting solution is placed in tray 15 frozen at -60°C and then heated for 96 hours pressure is gradually increased from freezing point to +25°C. It was dried in a lyophilizer by increasing the volume. In the final stage of drying, The highest vacuum achievable in the lyophilizer has been reapplied. A homogeneous white nonwoven sheet was obtained and then 3 × 4 cm 20 The samples were cut into specific dimensions. The thrombin concentration in the composition... It was 0.006 IU / cm². Example 2 Produced by Swedlight AB (Sweden), 25 with a molecular weight of 11.6×10⁵. g sodium hyaluronate obtained by biotechnological method, 2 L distilled water - 13 - dissolved in water and placed in a tray with a surface area of ​​1.478 cm² It was spilled. Later, the Hemophilia Patients Inter-Association Charity Renam Research and Production, Public Institution for Persons with Disabilities PG-9A thrombin, produced by the unit, for hemostasis tests. 5 reagent solution (with an activity of 18 IU) added to hyaluronate solution and is evenly distributed over its surface. The resulting solution is placed in the tray. frozen at -60°C and then heated for 96 hours pressure is gradually increased from freezing point to +25°C. It was dried in a lyophilizer by increasing the volume. In the final stage of drying, The highest vacuum achievable in the lyophilizer was reapplied. 10 A homogeneous white nonwoven sheet was obtained and then cut into 3 × 4 cm pieces. The samples were cut into specific dimensions. The thrombin concentration in the composition... It was 0.01 IU / cm². Example 3 15 Produced by Swedlight AB (Sweden), it has a molecular weight of 11.6 × 10⁵. g sodium hyaluronate obtained by biotechnological method, 2 L distilled water dissolved in water and placed in a tray with a surface area of ​​1.478 cm² It was spilled. Later, the Hemophilia Patients Inter-Association Charity Renam Research and Production 20, Public Institution for Persons with Disabilities PG-9A thrombin, produced by the unit, for hemostasis tests. The reagent solution (with an activity of 90 IU) was added to the hyaluronate solution. and is evenly distributed over its surface. The resulting solution is placed in the tray. frozen at -60°C and then heated for 96 hours The temperature is gradually increased from freezing point to +25°C while the pressure is 25°C. It was dried in a lyophilizer by increasing the volume. In the final stage of drying, The highest vacuum achievable in the lyophilizer has been reapplied. - 14 - A homogeneous white nonwoven sheet was obtained and then cut into 3 × 4 cm pieces. The samples were cut into specific dimensions. The thrombin concentration in the composition... It was 0.06 IU / cm². Example 4 5 The experiment was conducted on adult female rabbits. For the purpose of the experiment, Midline laparotomy is performed first to expose the liver margin. A liver scar model was created by applying this technique. Then, the parenchyma... and the liver capsule was incised and the compositions in Examples 1–3 were applied to the wound. It has been implemented. 10 The composition in Example 1 does not cause any visible changes in the wound. Dissolved without forming any hemostasis and within 3 minutes not observed. The compound in Example 2 initially adhered to the wound, but after 15 seconds It is almost completely dissolved within. Initiation of thrombogenesis. This was observed, and as a result, hemostasis was achieved after 1.5 minutes. The compound in Example 3 was stuck to the wound, not completely dissolved, and remained there for 15-20 minutes. Then hemostasis was achieved. Example 5 Produced by Swedlight AB (Sweden), it has a molecular weight of 11.6 × 10⁵. g sodium hyaluronate obtained by biotechnological methods with 0.2 g 25 - 15 - Benzildimethyl[3-(myristoylamino)propyl]ammonium chloride monohydrate, 2 L dissolved in distilled water and placed in a tray with a surface area of ​​1.478 cm² It was spilled. Later, the Hemophilia Patients Interassociation Charity Renam Research and Production Unit of the Public Institution for Persons with Disabilities PG-9A thrombin reagent 5, produced by [manufacturer name], for hemostasis testing. The solution (with an activity of 18 IU) was added to the hyaluronate solution and It is evenly distributed on the surface of the tray. The resulting solution is placed on the tray. frozen at -60°C and then heated for 96 hours pressure is gradually increased from freezing point to +25°C. The mixture was dried in a lyophilizer by increasing the volume. In the final stage of drying, 10 The highest vacuum achievable in the lyophilizer has been reapplied. A homogeneous white nonwoven sheet was obtained. The prepared sample conforms to article 1.2.4.0002.18 of the General Pharmacopoeia. It was used in the microbiological purity test carried out in accordance with the regulations. 15 (Source: State Pharmacopoeia of the Russian Federation, Edition IV, Volume 1, Pharmaceuticals) Analytical Methods, Statistical Analysis Methods, Microbiological Purity, (p. 1128). Microorganism strains representing various taxonomic bacterial groups 20 A strain representing yeast-like fungi was used. microorganisms; Pseudomonas representing Gram-negative bacteria aeruginosa and Escherichia coli, Gram-positive spore-forming bacteria Bacillus cereus, representing Gram-positive bacteria. Candida 25, representing Staphylococcus aureus and yeast-like fungi. It has become albicans. - 16 - The test results showed no growth of microorganisms. Example 6 Produced by Swedlight AB (Sweden), 5 with a molecular weight of 11.6×10⁵. 0.2 g sodium hyaluronate obtained by biotechnological methods Benzildimethyl[3-(myristoylamino)propyl]ammonium chloride monohydrate and 20 g dioxomethyltetrahydropyrimidine, dissolved in 2 L of distilled water and each It was poured into three separate trays with a surface area of ​​1,478 cm². Later, the Hemophilia Patients Inter-Association Charity for the Disabled Individuals are employed by the Renam Research and Production Unit of the Public Institution. PG-9A thrombin reagent solutions produced for hemostasis tests. (with activity levels of 9, 18 and 90 IU respectively) added to each tray and It is distributed homogeneously on the surface. The resulting solutions are placed in trays 15 frozen at -60°C and then heated for 96 hours pressure is gradually increased from freezing point to +25°C. It was dried in a lyophilizer by increasing the volume. In the final stage of drying, The highest vacuum achievable in the lyophilizer has been reapplied. Homogeneous white nonwoven sheets were obtained and then 3 × 4 cm 20 They were cut into samples of the specified dimensions. The obtained samples were then experimentally analyzed on a thigh wound model. It has been tested as such. Adult female rabbits were used in the experiment. Thigh to induce muscle injury and simulate significant bleeding, 25 a longitudinal incision involving skin, subcutaneous fat tissue, fascia, and muscle tissue - 17 - The procedure was performed. After the wound dressings were applied, the wound was left open for 3 days. It has been abandoned. The experimental results are given below. - Wound dressing with a composition having thrombin activity of 0.006 IU / cm² 5 It was completely dissolved and no hemostasis was observed within 3 minutes. Three days later, minimal signs of inflammation remained in the wound. Exudation has been detected. - In a composition with thrombin activity of 0.01 IU / cm², the wound dressing is completely... It dissolved and hemostasis was achieved after 2 minutes. Three days later, 10 in the wound Undissolved wound dressing remnants were detected. - In a compound with thrombin activity of 0.06 IU / cm², after 20 seconds Hemostasis has been achieved. Three days later, the wound is completely dry and there is no exudation. and showed no signs of inflammation, and the wound was not healed. Remnants of the covering have been identified. 15 - Example 7 Produced by Swedlight AB (Sweden), it has a molecular weight of 11.6 × 10⁵. 0.2 g sodium hyaluronate obtained by biotechnological methods Benzildimethyl[3-(myristoylamino)propyl]ammonium chloride monohydrate and 20 20 g lidocaine, dissolved in 2 L of distilled water and having a surface area of ​​1.478 cm² It was poured into a tray. Later, the Inter-Association of Hemophilia Patients Renam Research and Charity Organization for People with Disabilities PG-9A, produced by the Production Unit, for hemostasis testing. Thrombin reagent solution (with an activity of 90 IU) is added to hyaluronate solution 25 added and distributed homogeneously over the surface. The resulting solution, - 18 - frozen in a tray at -60°C and then heated for 96 hours. pressure is gradually increased from freezing point to +25°C. It was dried in a lyophilizer by increasing the volume. In the final stage of drying, The highest vacuum achievable in the lyophilizer has been reapplied. A homogeneous white nonwoven sheet was obtained. The aforementioned 5 Samples measuring 1 × 1 cm were cut from the plate. These specimens were taken from two adult males following the extraction of teeth number 38 and 48. This procedure was performed on the extraction socket to stop bleeding in the patient. In patients, bleeding stopped within 10 to 15 seconds. Three days later, 10 During the follow-up examination, the wound showed no signs of inflammation. The condition closed and it was observed that the patients did not require additional anesthesia. Example 8 Produced by Swedlight AB (Sweden), 15 with a molecular weight of 11.6×10⁵. Hemophilia with 1 g of biotechnologically produced sodium hyaluronate Patients, Inter-Association Charity, Disabled Persons, Public Institution Produced by Renam Research and Production Unit, for hemostasis tests. PG-9A thrombin reagent (with an activity of 16,200 IU), 0.2 L distilled water. dissolved in water and placed in a tray with a surface area of ​​148 cm² 20 The solution was poured out. The resulting solution was frozen in a tray at -60°C and Then, over 96 hours, the temperature gradually increased from freezing point to +25°C. It was dried in a lyophilizer by increasing the pressure while raising its volume. In the final stage of drying, the highest level achievable in the lyophilizer is... The vacuum has been reapplied. A dense, homogeneous white nonwoven fabric 25 - 19 - A layer was obtained. The thrombin concentration in the composition was 109 IU / cm². When the sheet was bent, it cracked and partially crumbled. A piece of material approximately 1 × 1 cm in size, Renam Research and 5 obtained from Control Plasma KM-2 produced by the Production Unit. It was crushed in a Petri dish containing 6 mL of plasma. Almost instantly. A clot has formed, but the material has not completely dissolved. Therefore, 109 At thrombin concentrations up to 1U / cm², the material is hemostatic. It retains its properties. The material remains in solid form, for example... It can be used in powder form in laparoscopic procedures. 10 Example 9 The sample obtained in Example 2 was analyzed using an ARL X'TRA X-ray diffraction system. It was examined using X-ray diffraction analysis (Figure). X-ray diffraction analysis revealed that the samples had a partially amorphous structure (XPRD). (characterized by the halo in the diffractogram) and between 24.8° and 43.8° theta It has a partially crystalline structure characterized by peaks around it. This has been shown. Brief description of the figures: "XPRD diffractogram of the sample according to the discovery" The figure titled [Figure Title] shows the XPRD diffractogram of the sample prepared in Example 2. It shows. - 20 - Therefore, the examples given demonstrate the rapid effect of the recommended hemostatic agent. It demonstrates that it has proven to be highly effective. The substance is used to stop bleeding in infected wounds or to treat these wounds. in various forms and in different ways, including preventing infection It can be used under various conditions and, thanks to its adhesive properties, 5 It is easy to implement. 15

Claims

- 21 - Requests 1. Contains a compound containing thrombin and hyaluronan in solid form. It is a hemostatic agent with a thrombin content of less than 0.01 IU / cm². It is characterized by its absence and not exceeding 109 IU / cm².

2. According to claim 1, it is a hemostatic agent, and hyaluronan is partially crystalline. It is characterized by being in a specific form.

3. According to Claim 1, it is a hemostatic agent and also an antiseptic and / or It is characterized by containing analgesic and / or wound-healing components. 20

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