Sponge for hemostasis

Abstract

The present invention relates to a sponge for hemostasis and a use thereof, the sponge comprising: thrombin; and at least one biomaterial selected from the group consisting of collagen, gelatin, fibrin, polysaccharides, synthetic biodegradable biomaterials, and derivatives thereof.

Description

hemostatic sponge

[0001] The present invention relates to a hemostatic sponge and its use.

[0002] Hemostatic agents and sealants are currently used as adjunctive tools to stop bleeding, including bleeding during surgery. The FDA has approved hemostatic matrices, such as FLOSEAL® (Baxter International), commonly used to enhance the natural coagulation cascade or mechanically stop bleeding at surgical or wound sites. FLOSEAL® is a hemostatic sealant containing human thrombin in a gelatin matrix.

[0003] Another hemostatic agent is Tachosil® (Baxter International), a collagen sponge matrix combined with the coagulation factors fibrinogen and thrombin. It is activated by blood and body fluids, forming a clot that adheres the sponge to the tissue surface, stanching the hemorrhage within minutes.

[0004] Other forms of hemostatic compositions include hemostatic sponges, pads, or powders containing plant-based materials, such as purified plant starches linked to carboxymethyl groups to modify the starch, such as Starch Medical. A hemostatic composition containing gelatinized potato starch is disclosed in U.S. Patent Publication No. 8,623,842.

[0005]

[0006] [Prior Art Literature]

[0007] [Patent Document]

[0008] (Patent Document 1) U.S. Patent Publication No. 8,623,842

[0009]

[0010] The present invention seeks to provide an improved hemostatic sponge.

[0011] The present invention also seeks to provide a use of a hemostatic sponge for treating damage at a site where a wound or bleeding has occurred.

[0012] The present invention relates to thrombin;

[0013] A hemostatic sponge is provided comprising at least one biomaterial selected from the group consisting of collagen, gelatin, fibrin, polysaccharides, synthetic biodegradable biomaterials, and derivatives thereof. The sponge may be in the form of a sheet or a bullet.

[0014] The above thrombin may be impregnated into a hemostatic sponge. In this case, impregnation means that thrombin is absorbed into the hemostatic sponge for the purpose of this invention.

[0015] Thrombin is an enzyme involved in the blood coagulation mechanism and has the property of hydrolyzing fibrinogen. The thrombin that can be used in the present invention is not particularly limited as long as it can be applied by impregnating it into a sponge. Examples of thrombin that can be used include thrombin listed in the 12th revised Korean Pharmacopoeia or human recombinant thrombin RECOTHROM® (Baxter).

[0016] The sponge used herein is a porous network of biomaterial capable of absorbing bodily fluids when applied to a wound site. This allows blood from the wound (including all blood components, such as blood cells or coagulation proteins) to flow into the sponge. Therefore, the porous sponge according to the present invention has an internal volume accessible to external fluids, such as blood, when applied to a patient.

[0017] The sponge may comprise one or more biomaterials usable in the art. The biomaterial may be, in particular, one or more selected from the group consisting of collagen, gelatin, particularly cross-linked gelatin, fibrin, polysaccharides, particularly chitosan, oxidized cellulose, aldehyde-activated dextrans, starch-based polyaldehydes, synthetic biodegradable materials, particularly polylactic acid or polyglycolic acid, and derivatives thereof. The biomaterial is preferably gelatin or collagen.

[0018] The above sponge may additionally contain albumin.

[0019] In one embodiment, "gelatin sponge" refers to gelatin processed into a sponge-like structure with a porous structure. The gelatin that can be used as a raw material is not particularly limited as long as it is suitable for use as a pharmaceutical product. Animal-derived gelatin, for example, medical gelatin manufactured from cow bones, pig skin, etc., can be used. Processing methods include, for example, foaming a gelatin solution and freeze-drying the resulting foam.

[0020] In another embodiment, the "collagen sponge" may be prepared by lyophilization, i.e., freeze-drying of a collagen gel, suspension, or solution (whereas conventional air drying produces a collagen film). The collagen used in the present invention may be from any collagen suitable for forming a gel, including materials from liquid, pasty, fibrous, or powdery collagenous materials that can be processed into a porous or fibrous matrix. The preparation of collagen gels for producing sponges is described, for example, in EP 0891193, and may involve acidification and subsequent pH neutralization until gel formation occurs. To improve gel-forming ability or solubility, the collagen may be partially or fully hydrolyzed or modified, provided that its ability to form a stable sponge when dried is not impaired.

[0021] The collagen sponge according to the present invention preferably has a lower density than the density of the collagen film. Preferably, the density is about 5 to 100 mg / cm 3 On the other hand, the density of the film is about 650 mg / cm 3 It is excessive. For example, the collagen sponge may be commercially available under the trade name Matristypt®.

[0022] In this application, the term "sheet" means a material that can be a base for fixing an object, shape, or other material having a certain thickness and an area that can be folded or rounded. When applied to a bleeding site, hemostasis is achieved by absorbing or fixing exuding or flowing blood. The total size may be appropriately determined, but a size that is easy to use in an actual clinical setting, for example, a rectangular sheet having a length of 8.0 mm and a width of 12.0 mm, a square sheet having a length and a width of 20.0 mm each, a rectangular sheet having a length of 10.0 mm and a width of 20.0 mm, etc. may be appropriately used. In addition, it may be cut to a suitable size at the time of use. Alternatively, the sheet may have various sizes, such as 10 X 10 mm, 20 X 20 mm, 10 X 20 mm, 20 X 20 mm, 40 X 40 mm, 50 X 50 mm, and 100 X 100 mm.

[0023] In one embodiment, the sheet-shaped hemostatic sponge of the present invention, i.e., the hemostatic sheet, may have a uniform thickness in a range of 0.1 to 5.0 mm, for example, 0.5 to 4.0 mm, 1.0 to 3.5 mm, or 2.0 to 3.0 mm, and the upper and lower limits may be applied in combination with each other, and specifically, may have a thickness of 0.1 to 0.5 mm. Since the thickness may show some variation depending on the manufacturing method of the sheet, a variation of + / - 10% from the upper and lower limits may be recognized as an acceptable value.

[0024] In one embodiment, the hemostatic sheet of the present invention comprises 5 to 100 mg / cm 3 , for example, 10 to 70 mg / cm 3 , 20 to 60 mg / cm 3 , 25 to 55 mg / cm 3 , 30 to 55 mg / cm 3 , 35 to 50 mg / cm 3 , or 40 to 50 mg / cm 3 The density can vary within a range, and upper and lower limits can be combined. Since the sheet is applied to a wound or bleeding area, the density can vary within a certain range. For example, a variation of + / - 10% from the upper and lower limits may be considered acceptable. If the density is too high, cracks may occur due to deformation, and a certain density is required to maintain shape when wet. Therefore, it will be necessary to set an appropriate density.

[0025] In another embodiment, the hemostatic sponge of the present invention may have a bullet-shaped shape, wherein the bullet-shaped sponge may have a diameter of 8 to 15 mm and a height of 25 mm.

[0026] When an external fluid, for example blood, comes into contact with the sponge according to the present invention, for example, one or more polymers, for example hydrophilic polymer components comprising reactive groups, may react with blood components and / or the surface of the biomaterial matrix to crosslink those components that bind to the reactive groups.

[0027] Accordingly, the sponge of the present invention may additionally comprise one or more polymers. The polymer may be a hydrophilic polymer, and the hydrophilic polymer component may be a hydrophilic cross-linking agent capable of causing cross-linking through a reaction with a reactive group when the sponge is applied to a wound or bleeding site of a patient requiring hemostasis.

[0028] In one embodiment, the hydrophilic polymer component is a polyalkylene oxide polymer, such as PEG. The polymer may include electrophilic groups, such as amino groups, carboxyl groups, thiol groups, and hydroxyl groups of proteins, or mixtures thereof.

[0029] Examples of amino-specific reactive groups include NHS-ester groups, imidoester groups, aldehyde groups, carboxyl groups in the presence of carbodiimides, isocyanates, or THPP (beta-[tris(hydroxymethyl)phosphino] propionic acid), and more specifically, pentaerythritol poly(ethylene glycol) ether tetrasuccinimidyl glutarate (= pentaerythritol tetrakis[1-1'-oxo-5'-succinimidylpentanoate-2-poly-oxoethylene glycol] ether (= NHS-PEG with MW 10,000).

[0030] Examples of carboxyl-specific reactive groups include amino groups in the presence of carbodiimides.

[0031] Examples of thiol-specific reactive groups include maleimide or haloacetyl.

[0032] Examples of hydroxyl-specific reactive groups include isocyanate groups.

[0033] In one embodiment, the hemostatic sponge according to the present invention may comprise additional physiological substances. For example, the sheet may comprise an antifibrinolytic agent, such as a plasminogen activator inhibitor, a plasmin inhibitor, or a fibrinolytic agent inactivator.

[0034] Additionally, the hemostatic sponge may further comprise additional pharmacologically active substances, such as antibiotics, antifungal agents, growth factors, analgesics, etc. In addition, the sponge may comprise additional substances that affect blood clotting rate, hemostasis, and sealing qualities, such as tensile strength, internal adhesive strength, and durability.

[0035] In one embodiment, the sponge of the present invention may exhibit hemostatic activity at a wound or bleeding site, thereby treating the injury. More specifically, the sponge may be used to control hemostasis during surgical or abdominal surgery, such as during dental surgery.

[0036] When using the sponge of the present invention, a blood coagulation effect, i.e., a hemostatic effect, can be observed within about 1 to 2 minutes. Preferably, the hemostatic effect can be observed within 30 seconds.

[0037] In one embodiment, the sponge of the present invention may be prepared from a composition comprising thrombin and a biomaterial. In one specific example, the sponge may be prepared in a freeze-dried form using a composition comprising thrombin, collagen, and albumin. A specific method for preparing a hemostatic sponge from the composition may utilize a method known in the art.

[0038] The above composition may contain collagen in an amount of 1 to 10% (w / v), for example, 2% (w / v). While an increase in the collagen content increases the hemostatic effect, it may also decrease economic feasibility from a commercial perspective. The above-mentioned content may be appropriately determined by considering both the hemostatic effect and economic feasibility.

[0039] The composition may also comprise an effective amount of thrombin. The term "effective amount" as used herein means an amount of thrombin sufficient to exhibit hemostatic activity. For example, the thrombin may be comprised in an amount of 100 to 2500 IU / mL, such as 250 IU to 2500 IU / mL, such as 100, 200, 250, 300, 400, or 500 IU / mL, and more specifically, 100, 200, 250, or 500 IU / mL, preferably 250 or 500 IU / mL.

[0040] The composition may comprise albumin in an amount of 0.1 to 5% (w / v), for example 0.5% (w / v).

[0041] In one specific embodiment, the composition comprises

[0042] Collagen 2% (w / v);

[0043] Thrombin 100, 200, 250, or 500 IU / mL; and

[0044] Albumin 0.5% (w / v)

[0045] Includes.

[0046] The above composition may also contain additional ingredients as described above.

[0047] In another embodiment, the present invention provides a kit comprising the hemostatic sponge; a buffer solution; and instructions for use.

[0048]

[0049] The hemostatic sponge of the present invention exhibits rapid and effective hemostatic properties when applied to a bleeding site. Accordingly, it can be effectively used in areas prone to bleeding, such as during surgical procedures.

[0050]

[0051] Figure 1 is a photograph of a sheet-shaped hemostatic agent containing thrombin.

[0052] Figure 2 is a photograph showing the manufacturing process of a liver damage model.

[0053] Figure 3 is a photograph showing the hemostatic effect in an animal model of liver damage.

[0054] Figure 4 is a graph showing each hemostasis time in an animal model of liver injury.

[0055]

[0056] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples, in accordance with the gist of the present invention.

[0057]

[0058] Confirmation of the hemostatic effect of thrombin-containing hemostatic sponge in a liver injury model.

[0059] I. Materials and Methods

[0060] 1. Experimental materials

[0061] Saline (0.9% NaCl) was used as a negative control in this experiment. As an experimental group, the sheet-shaped hemostatic agent containing thrombin of the present invention was used in three concentrations (TH1, TH2, TH3).

[0062] As experimental materials, 6% hydrated collagen (OSSCORE, Denhouse), thrombin (MP Bio, Leeyeon Pharmaceutical), and albumin (Green Cross) were purchased and used in the following experiments. 2% (w / v) collagen, 100, 200, or 500 IU / mL thrombin, and 0.5% albumin were mixed per 1 mL of hemostatic mixture and lyophilized into a sheet-shaped sponge.

[0063]

[0064] 2. Composition of experimental animals and experimental groups

[0065] In this experiment, 8-week-old male Sprague Dawley rats (SD rats) were purchased from Sam Taco (Osan, Korea) and used after a one-week acclimation period. The average weight of SD rats was 180–200 g. The experimental animal housing room was maintained at a 12-hour light / dark cycle, a temperature of 23±2°C, and a humidity of 50–60%. This animal experiment was conducted in compliance with the Guide for Animal Experimentation of Wonkwang University (Approval No. WKU 21-30).

[0066] The experimental animals were divided into four groups. The negative control group (Sham) received saline, and the experimental group (TH1) received a sheet-shaped hemostatic agent containing experimental thrombin (Table 1). Four animals were used in each group.

[0067]

[0068] Group Treatment n Liver Injury Substance used Dosage 1 Sham Yes Saline (0.9% NaCl) 4 2 TH 1 Yes Thrombin-containing sheet-type hemostatic agent 100 IU 4 3 TH 2 Yes Thrombin-containing sheet-type hemostatic agent 200 IU 4 4 TH 3 Yes Thrombin-containing sheet-type hemostatic agent 500 IU 4

[0069]

[0070] 3. Creation of a liver injury model and measurement of hemostatic effects

[0071] To create a liver injury model, after animal acclimatization, general anesthesia was performed with isoflurane, the hair on the abdomen was removed, and the peritoneum was incised to expose the liver. A 4 mm wound was made on the left lobe of the exposed liver, and when bleeding started, a sample (2 X 2 cm 2 ) was introduced to measure the time until hemostasis was completed (Fig. 2).

[0072]

[0073] 4. Statistical processing

[0074] The results obtained in this experiment are expressed as the mean ± standard deviation (mean ± SD). Statistical analysis of the significance of each experimental group was performed using one-way Anova SPSS v.12 for statistical comparison between the control and experimental groups. Duncan's post hoc test was performed, and a value of (p < 0.05) or less was considered statistically significant.

[0075]

[0076] II. Results and Discussion

[0077] 1. Blood clotting time measurement results

[0078] The results of measuring the hemostasis time of the sheet-shaped hemostatic agent of the present invention using a liver injury animal model are shown in Table 2.

[0079]

[0080] GroupBlood clotting time (sec)Sham107.5 ± 14.5TH134.5 ± 7.9TH223.5 ± 1.7TH313.3 ± 3.9Each value is Mean ± SD.

[0081]

[0082] In a liver injury model, the sheet-shaped hemostatic agent of the present invention exhibited a hemostatic effect as blood reacted with the hemostatic sheet after the initial bleeding, and a rapid hemostatic effect was observed in a thrombin concentration-dependent manner (Figs. 3 and 4). In particular, based on the above experimental results, it is thought that the hemostatic effect will be excellent when thrombin is 250 IU / mL or higher.

Claims

1. Thrombin; One or more biomaterials selected from the group consisting of collagen, gelatin, fibrin, polysaccharides, synthetic biodegradable biomaterials, and derivatives thereof. A hemostatic sponge containing:

2. A hemostatic sponge according to claim 1, wherein the biomaterial is gelatin or collagen.

3. A hemostatic sponge according to claim 1, wherein the sponge further comprises albumin.

4. A hemostatic sponge according to claim 1, wherein the sponge is sheet-shaped or bullet-shaped.

5. A hemostatic sponge manufactured in a lyophilized form using a composition comprising thrombin, collagen, and albumin according to claim 1.

6. A hemostatic sponge according to claim 5, wherein the composition comprises collagen in an amount of 2% (w / v).

7. A hemostatic sponge according to claim 5, wherein the composition contains thrombin in an amount of 100 to 2500 IU / mL.

8. A hemostatic sponge according to claim 5, wherein the composition contains thrombin in an amount of 250 to 1000 IU / mL.

9. A hemostatic sponge according to claim 5, wherein the composition comprises albumin in an amount of 0.5% (w / v).

10. In the fifth paragraph, the composition Collagen 2% (w / v) Thrombin 100, 200, 250, or 500 IU / mL; and Albumin 0.5% (w / v) A hemostatic sponge comprising:

11. In the first paragraph, the hemostatic sponge is sheet-shaped and has a density of 5 to 100 mg / cm 3 A hemostatic sponge having a density of 0.1 to 5.0 mm and a thickness of 0.1 to 5.0 mm.

12. A hemostatic sponge according to claim 1, wherein the sponge exhibits hemostatic activity at a site where a wound or bleeding has occurred.

13. A hemostatic sponge according to claim 1, wherein the sponge is used for hemostasis during surgical operations.

14. A hemostatic sponge according to claim 1, wherein the sponge is used for hemostasis during dental surgery.

15. A kit comprising a hemostatic sponge of paragraph 1; a buffer solution; and instructions for use.

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