Hemostatic composition and hemostatic kit comprising same

Abstract

The present invention relates to a hemostatic composition and a kit comprising same. The hemostatic composition according to the present invention comprises a polysaccharide, a stabilizer, a binder, thrombin and an antibiotic, wherein the polysaccharide is coated with the stabilizer, the binder and thrombin, and the antibiotic is mixed with the coated polysaccharide. The hemostatic composition according to the present invention is in the form of a polysaccharide powder coated with the stabilizer, the binder, and thrombin, and thus can be stored for a long period of time even at room temperature by having a high interaction area between the stabilizer and thrombin, can be properly fixed to an application site by means of the binder, and has an excellent hemostatic effect by having a high surface area of thrombin, thereby being useful as a hemostatic agent. In addition, since an antibiotic is included, infections that can occur at a bleeding site can be prevented.

Description

Hemostatic composition and hemostatic kit containing the same

[0001] The present invention relates to a hemostatic composition and a kit including the same, and more particularly, to a hemostatic composition that can be rapidly applied and a kit including the same.

[0002] In the medical field, controlling bleeding is essential and crucial during surgical procedures to minimize blood loss, reduce postoperative complications, and shorten operating time. When blood, which is directly related to human life, leaks out of the body due to damage to a living blood vessel, various coagulation factors act as enzymes, and substances such as calcium ions and thromboplastin interact to ultimately form a fibrin network.

[0003] Blood cells in the blood adhere to and coagulate within the formed fibrin meshwork, thereby sealing damaged blood vessels and achieving hemostasis. During this process, thrombin acts on fibrinogen, cleaving the fibrinogen peptide to produce fibrin. The electrostatic attraction between the fibrin molecules forms a network. Fibrinogen itself has no hemostatic effect, but its hemostatic effect is enhanced when combined with thrombin. In other words, thrombin plays a crucial role in the body's hemostatic response.

[0004] However, in the case of hemostasis through the body's hemostatic mechanism, it can sufficiently have a hemostatic effect for minor bleeding such as capillary bleeding due to minor wounds, but in the case of excessive bleeding that may occur during surgical operations or emergency situations, it has limitations in sufficiently stopping the bleeding, so the use of other effective hemostatic methods is required. These hemostatic methods are largely divided into mechanical methods, methods using heat and cold, and chemical methods. Mechanical and heat and cold methods can affect the tissue at the bleeding site and its surroundings, causing secondary damage. Chemical methods use biological hemostatic substances such as thrombin and fibrinogen, or drugs such as vasoconstrictors. In particular, biological hemostatic substances such as thrombin and fibrinogen are suitable as hemostatic substances because they do not cause effects such as tissue damage since they are components that already exist in the body.

[0005] A key function of hemostatic agents is to rapidly coagulate blood, preventing excessive bleeding in various situations such as wounds and surgical procedures. A variety of drugs, polymers, and proteins are currently used as hemostatic agents, among which thrombin preparations exhibit excellent hemostatic effects.

[0006] Conventional thrombin preparations, available in liquid or powder form, often flowed down the bleeding site, failing to fully exert their hemostatic effects. To overcome this problem, WO1990 / 013320 disclosed a sheet immobilizing thrombin on a bioabsorbable material. However, the sheet-like formulation failed to exhibit sufficient hemostatic effects, and its manufacturing difficulties and practical application challenges prevented its commercialization.

[0007] In addition, there are gel-type hemostatic agents that mix bovine thrombin with bovine gelatin or porcine collagen, but these types of hemostatic agents carry the risk of infection such as BSE (bovine spongiform encephalopathy) due to the addition of animal-derived polymers (gelatin, collagen), and in particular, collagen is a component that exists in body tissues and has a slow decomposition rate, so there is a problem of causing an inflammatory response following organ transplantation or causing adhesion between tissues.

[0008] Oxidized regenerated cellulose (ORC), which is not derived from animals and is widely used as a topical dressing in various conventional surgical procedures, is not suitable as an additive because the acidic residues present in cellulose denature the activity of thrombin when combined with thrombin, causing thrombin to lose its function.

[0009] Furthermore, thrombin, an enzyme isolated from blood, is unstable at room temperature, making long-term storage difficult. Typically, thrombin is lyophilized and then frozen for long-term storage. However, its activity deteriorates rapidly in solution, requiring the storage of thrombin to be removed immediately before use, which presents an inconvenience.

[0010] Another known hemostatic agent is fibrin adhesive, which combines thrombin with other coagulation factors. Fibrin adhesive, primarily composed of thrombin and fibrinogen, is a biotissue adhesive that utilizes the conversion of fibrinogen to fibrin by thrombin. Its purpose is adhesion, hemostasis, and closure. However, fibrinogen-containing biotissue adhesives require thawing after being frozen, making them inconvenient for use during emergency surgical procedures.

[0011] In order to solve the above-mentioned problems, a hemostatic agent is required that does not cause denaturation of thrombin when mixed, prevents thrombin from flowing out when applied to the body, contains a biocompatible and biodegradable polymer that can minimize infectious factors due to the use of animal-derived polymers, can be stored for a long period of time at room temperature, and can sufficiently exhibit a hemostatic effect.

[0012] Accordingly, the inventors of the present invention have made efforts to solve the above problems and have confirmed that when a hemostatic composition comprising a polysaccharide coated with a mixture of a stabilizer, a binder and thrombin and an antibiotic is used, long-term storage at room temperature is possible and this can be used as a powder-type hemostatic kit or a hydrogel-type hemostatic kit with excellent hemostatic effect, thereby completing the present invention.

[0013] The purpose of the present invention is to provide a hemostatic composition that can be stored for a long time at room temperature, can fix thrombin to a bleeding site or apply it to a wide area, and includes additives that minimize the risk of infection, and a hemostatic kit comprising the same.

[0014] To achieve the above object, the present invention provides a hemostatic composition comprising (a) a polysaccharide coated with a mixture of a stabilizer, a binder, and thrombin; and (b) an antibiotic.

[0015] The present invention also provides a hemostatic kit in powder form comprising (a) a hemostatic composition comprising a polysaccharide coated with a mixture of a stabilizer, a binder and thrombin and (b) an antibiotic; and a cationic substance.

[0016] The present invention also provides an endoscopic powder-type hemostatic kit comprising the powder-type hemostatic kit and an endoscopic spraying device.

[0017] The present invention also provides a hydrogel-type hemostatic kit comprising: (a) a first container containing a hemostatic composition comprising a polysaccharide coated with a mixture of a stabilizer, a binder, and thrombin, and (b) an antibiotic; a second container containing a cationic solution; and a connector connecting the first container and the second container.

[0018] The hemostatic composition according to the present invention is in the form of a polysaccharide powder coated with a stabilizer, a binder, and thrombin, and thus can be stored for a long time even at room temperature due to the high interaction area of ​​the stabilizer and thrombin. In addition, the binder allows it to be well fixed to the application site, and the high surface area of ​​thrombin provides an excellent hemostatic effect, making it useful as a hemostatic agent. In addition, since it contains an antibiotic, it can prevent infection that may occur at the bleeding site. In addition, it can be used as a hydrogel type by mixing it with a separately stored cationic solution, if necessary. In addition, the powdered hemostatic composition can be connected to an endoscopic spray device, if necessary, to suppress bleeding that occurs during endoscopic procedures, and thus can be applied to various bleeding situations.

[0019] Figure 1 is a photograph of a container storing a hemostatic composition in powder form according to the present invention.

[0020] Figure 2 is a photograph of an endoscopic powder-type hemostatic kit according to the present invention.

[0021] Figure 3 is a photograph of a hydrogel-type hemostatic kit according to the present invention.

[0022] Figure 4 is an SEM photograph of a powdered hemostatic composition and hydrogel according to the present invention.

[0023] In the present invention, a hemostatic composition comprising a polysaccharide coated with a mixture of a stabilizer, a binder, and thrombin; and an antibiotic in powder form can be stored for a long time at room temperature, and (a) when used with a cationic substance, it contributes to inducing hemostasis together with thrombin in powder form, can be easily applied to a wide area, and can fix the hemostatic agent to the application site due to the binder, and (b) when used with a cationic solution, it can be applied to the exact location of a local area in the form of a hydrogel, and does not flow down from the hemostatic site, thereby increasing the fixing power of the hemostatic agent.

[0024] In the present invention, a powdered hemostatic composition was prepared by coating a polysaccharide with various types of stabilizers, binders, and thrombin in specific contents, and including an antibiotic, and (a) a powdered hemostatic kit was prepared by mixing cationic substances, and (b) a hydrogel-type hemostatic kit was prepared by mixing cationic solutions, and their hemostatic ability, toxicity, long-term storage effect, antibacterial activity, etc. were evaluated.

[0025] As a result, it was confirmed that the hemostatic kit according to the present invention, in which the hemostatic composition and the cationic solution are stored separately, can be stored for a long time even at room temperature, has an excellent hemostatic effect without toxicity, and has adhesive strength and antibacterial properties that inhibit bacterial growth.

[0026] Accordingly, the present invention relates, in one aspect, to a hemostatic composition comprising (a) a polysaccharide coated with a mixture of a stabilizer, a binder and thrombin; and (b) an antibiotic.

[0027] In the present invention, the stabilizer stabilizes the activity of thrombin to facilitate storage, and examples thereof include serum albumin, mannitol, sorbitol, sucrose, trehalose, dextran, glucose, sodium acetate, glycine, etc., and mannitol or sorbitol is preferable.

[0028] In the present invention, the binder is used to make the hemostatic composition well fixed to the bleeding site by contacting the tissue, and examples thereof include polypyrrolidone (PVP), polyethylene glycol (PEG), polyethylene oxide (PEO), etc., and it is preferable to use polypyrrolidone.

[0029] In the present invention, examples of the polysaccharide include starch, alginate, pullulan, carboxymethyl cellulose, dextrin, etc., and starch is preferable, carboxymethyl starch is more preferable, and carboxymethyl starch having a molecular weight of 500,000 to 1,000,000 g / mol is even more preferable.

[0030] In the present invention, the antibiotic has antibacterial properties to prevent infection that may occur at the bleeding site, and examples thereof include gentamicin, penicillin, cephalosporin, monobactam, and carbapenem, with gentamicin being preferable.

[0031] The hemostatic composition of the present invention comprises a polysaccharide coated with a mixture of a stabilizer, a binder, and thrombin, which comprises 1 to 45 wt% of the stabilizer, 2 to 10 wt% of the binder, 0.01 to 10 wt% of the thrombin, and 35 to 96.99 wt% of the polysaccharide, and the antibiotic is preferably contained in an amount of 0.5 to 1.5 wt% based on 100 wt% of the polysaccharide coated with the mixture of a stabilizer, a binder, and thrombin.

[0032] If the above stabilizer is less than 1 wt%, there is a problem that stabilization of thrombin is not achieved, making storage difficult. If it exceeds 45 wt%, there is a problem that a hydrogel is not properly formed when mixed with a cationic solution, and a physical film is not formed at the bleeding site when used alone.

[0033] If the above binder is less than 2 wt%, the adhesive strength is not properly implemented, and if it exceeds 10 wt%, it may interfere with the blood contact of the polysaccharide, thereby interfering with the hemostatic effect.

[0034] When the above thrombin is less than 0.01 wt%, the hemostatic effect is minimal, and when it exceeds 10 wt%, there is an economic problem as only the manufacturing cost increases without any improvement in the hemostatic effect.

[0035] When the content of the above polysaccharide is less than 35 wt%, there is a problem that a hydrogel is not properly formed when mixed with a cationic solution, and a physical film is not formed at the bleeding site when used alone. When it exceeds 96.99 wt%, there is a problem that proper mixing with a stabilizer or thrombin is difficult.

[0036] If the amount of the above antibiotic is less than 0.5 parts by weight relative to 100 parts by weight of the polysaccharide coated with the mixture of stabilizer, binder and thrombin, the antibacterial effect is not properly realized, and if it exceeds 1.5 parts by weight, there is no additional improvement in the antibacterial effect, and on the contrary, there is a problem of toxicity.

[0037] In the present invention, a polysaccharide coated with a mixture of a stabilizer, a binder, and thrombin can be prepared by a method comprising the steps of (a) dissolving the stabilizer, the binder, and the thrombin in distilled water and homogenizing; (b) immersing the polysaccharide in the homogenization solution; (c) separating the immersed polysaccharide from the solution; and (d) lyophilizing and pulverizing the separated polysaccharide.

[0038] The hemostatic composition comprising (a) a polysaccharide coated with a mixture of a stabilizer, a binder, and thrombin; and (b) an antibiotic of the present invention is in powder form, and thus can be stored for a long period of time even at room temperature.

[0039] Meanwhile, the present invention relates to a hemostatic composition comprising (a) a polysaccharide coated with a mixture of a stabilizer, a binder and thrombin and (b) an antibiotic; and a powder-type hemostatic kit comprising a cationic substance.

[0040] In the present invention, a powder-type hemostatic kit means a hemostatic composition and a cationic substance stored in a container.

[0041] The hemostatic composition is the same as described above, and the cationic substance is for assisting the activity of thrombin, and examples thereof include calcium chloride (CaCl2), sodium chloride (NaCl), and mixtures thereof, with calcium chloride (CaCl2) being preferred.

[0042] In the present invention, it is preferable that the cationic substance be included in an amount of 0.01 to 5 parts by weight per 100 parts by weight of the hemostatic composition. If the content of the cationic substance is less than 0.01 parts by weight, there is a problem in that it does not sufficiently contribute to the activity of thrombin, and if it exceeds 5 parts by weight, there is no improvement in thrombin activity due to the excess, which is economically undesirable.

[0043] In the present invention, the hemostatic composition and the cationic substance can be stored in a container. As long as the hemostatic composition and the cationic substance can be stored, the material, shape, etc. of the container can be used without any special restrictions. In the case of powder form, it is preferable to use a bellows-shaped container.

[0044] More specifically, as illustrated in FIG. 1, the bellows-shaped container comprises a container capable of storing contents and a screw cap fastened to an inlet at one end of the container through which the contents are discharged. The bellows-shaped container preferably utilizes a structure and material capable of being compressed and restored to spray powder through the inlet. The container may be exemplified by the multipurpose medical powder sprayer disclosed in Korean Patent No. 10-2391900.

[0045] The above powder-type hemostatic kit can be used immediately in situations requiring the use of a hemostatic agent, enabling an immediate response in emergency situations.

[0046] Meanwhile, the present invention sought to confirm that a powdered hemostatic composition can be used to suppress bleeding occurring during endoscopic procedures by using it in conjunction with an endoscopic spray device. In the present invention, a powdered hemostatic kit was used in conjunction with an endoscopic spray device to evaluate its hemostatic performance during endoscopic procedures.

[0047] As a result, it was found that the powder-type hemostatic kit of the present invention can be administered through an endoscopic spray device and has an excellent hemostatic effect.

[0048] Accordingly, the present invention, from another aspect, relates to an endoscopic powder hemostatic kit comprising a powder hemostatic kit and an endoscopic spraying device.

[0049] The endoscopic spray device according to the present invention can be used without any special restrictions as long as it can spray a powdered hemostatic agent during surgical operations.

[0050] As illustrated in FIG. 2, the endoscopic spray device may include a container coupling portion to which a powder-type hemostatic kit can be connected, a tube coupling portion to which a tube passing through the endoscopic device is coupled, a motor portion for supplying external air to the powder-type hemostatic kit and the tube, a power supply portion for supplying power to the motor, and a switch portion for controlling the operation of the motor. The endoscopic spray device may be exemplified by a medical powder spray device capable of easily controlling the flow rate of the sprayed powder disclosed in Korean Patent No. 10-2443853.

[0051] Meanwhile, the present invention relates to a hydrogel-type hemostatic kit comprising, from another aspect, (a) a first container comprising a hemostatic composition comprising a polysaccharide coated with a mixture of a stabilizer, a binder, and thrombin, and (b) an antibiotic; a second container comprising a cationic solution; and a connector connecting the first container and the second container.

[0052] The hemostatic composition is the same as described above, and the cationic solution is mixed with the hemostatic composition to form a hydrogel, and examples thereof include calcium chloride (CaCl2) solution, sodium chloride (NaCl) solution, and mixtures thereof, and a calcium chloride (CaCl2) solution is preferred. The cationic solution can be prepared by adding a water-soluble solvent to 0.01 to 5 parts by weight of a cationic substance per 100 parts by weight of the hemostatic composition on a dry powder basis.

[0053] The first container constituting the hydrogel-type hemostatic kit of the present invention can be used without any special restrictions on the material, shape, etc. of the container as long as it can store the hemostatic composition and the second container can store the cationic solution, but it is preferable to use a commonly used syringe-type container.

[0054] More specifically, as illustrated in FIG. 3, the syringe-shaped container is composed of a container capable of storing contents, a screw cap fastened to an inlet through which contents are discharged at one end of the container, a plunger inserted into the interior of the container and reciprocally moved in a linear manner by a physical force, and a piston coupled to one end of the plunger and positioned inside the container.

[0055] The connector is preferably in the form of a hollow tube that connects a portion of the first container and a portion of the second container so that the positive ion solution of the second container can be transferred to the hemostatic composition of the first container, and then the mixed positive ion solution and hemostatic composition can be transferred to the second container.

[0056] The hemostatic composition in powder form in the first container forms a hydrogel when mixed with a cationic solution, and the formed hydrogel has an average pore size of 100 μm to 1000 μm. If the average pore size of the hydrogel is less than 100 μm, the blood absorption rate is reduced, and if it exceeds 1000 μm, there is a concern that the hemostatic effect is reduced because the hydrogel cannot maintain its shape after absorbing blood.

[0057] The formation time of the above hydrogel is 5 seconds to 3 minutes, which can prevent serious ligation and bleeding in emergency situations.

[0058] [Example]

[0059] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.

[0060] Examples 1-5: Preparation of hemostatic composition

[0061] The stabilizer (mannitol, sorbitol), binder (polypyrrolidone), and thrombin were added to 10 ml of distilled water according to the composition shown in Table 1 below, dissolved by a physical method (impeller, etc.) at room temperature, homogenized, and then polysaccharide (carboxymethyl starch, molecular weight: 500,000 to 1,000,000 g / mol) was immersed in the homogenized solution. The immersed polysaccharide (carboxymethyl starch) was separated from the excess solution by passing it through a filter (45 μm or less), and then the separated polysaccharide (carboxymethyl starch) was freeze-dried and pulverized to prepare a polysaccharide coated with a mixture of stabilizer, binder, and thrombin, and an antibiotic was mixed therein to prepare a hemostatic composition.

[0062] Polysaccharide antibiotics coated with a mixture of weight (%) stabilizer, binder and thrombin Carboxymethyl starch Mannitol Sorbitol Thrombin Polypyrrolidone Gentamicin Example 16527-351.5 Example 265-27350.5 Example 36530-0.014.991.5 Example 43545-10100.5 Example 5961-120.5

[0063]

[0064] Comparative Examples 1 to 13: Preparation of a hemostatic composition

[0065] The stabilizer (mannitol, sorbitol, collagen, gelatin), binder (polypyrrolidone), and thrombin were added to 10 ml of distilled water according to the composition shown in Table 2 below, dissolved by a physical method (impeller, etc.), homogenized, and then polysaccharide (carboxymethyl starch) was immersed in the homogenized solution. The immersed polysaccharide (carboxymethyl starch) was separated from the excess solution by passing it through a filter (45 μm or less), and then the separated polysaccharide (carboxymethyl starch) was freeze-dried and pulverized to prepare a polysaccharide coated with a mixture of stabilizer, binder, and thrombin, and an antibiotic was mixed therein to prepare a hemostatic composition.

[0066] Polysaccharide antibiotics coated with a mixture of weight (%) stabilizer, binder and thrombin Carboxymethyl starch Collagen Gelatin Mannitol Sorbitol Thrombin Polypyrrolidone Gentamicin Comparative Example 1-65-27-35 1.5 Comparative Example 2--65 15-15 51.5 Comparative Example 3-65-32-3-0.5 Comparative Example 4--65 32-3-0.5 Comparative Example 565--30--51.5 Comparative Example 665---30-50.5 Comparative Example 765--32-3-2 Comparative Example 865---32 3-1.5 Comparative Example 970--29-0.01 0.99 0.5 Comparative Example 1030--65-32 1.5 Comparative Example 1155--4.99-0.01401.5Comparative example 1297--0.95-0.00520.5Comparative example 1386.95--0.05-3100.4

[0067]

[0068] Manufacturing Examples 1 to 5: Manufacturing of a powder-type hemostatic kit

[0069] To manufacture a powder-type hemostatic kit, 0.5 to 1 g of the hemostatic composition manufactured in Examples 1 to 5 and 0.025 to 0.05 g of calcium chloride were filled into a bellows-shaped container.

[0070] Manufacturing Examples 6 to 18: Manufacturing of a powder-type hemostatic kit

[0071] To manufacture a powder-type hemostatic kit, 0.5 to 1 g of the hemostatic composition manufactured in Comparative Examples 1 to 13 and 0.025 to 0.05 g of calcium chloride were filled into a bellows-shaped container.

[0072] Manufacturing Examples 19-23: Manufacturing of a powder-type hemostatic kit

[0073] A powder-type hemostatic kit was manufactured by filling 0.5 to 1 g of a hemostatic composition containing a stabilizer, binder, thrombin, polysaccharide, and antibiotic having the same composition as Examples 1 to 5, except that the stabilizer, binder, and thrombin were not coated on the polysaccharide, and 0.025 to 0.05 g of calcium chloride into a bellows-shaped container.

[0074] Manufacturing Examples 24 to 36: Manufacturing of a powder-type hemostatic kit

[0075] A powder-type hemostatic kit was manufactured by filling 0.5 to 1 g of a hemostatic composition containing a stabilizer, binder, thrombin, polysaccharide, and antibiotic having the same composition as Comparative Examples 1 to 13, except that the stabilizer, binder, and thrombin were not coated on the polysaccharide, and 0.025 to 0.05 g of calcium chloride into a bellows-shaped container.

[0076] Manufacturing Examples 37 to 41: Manufacturing of a hydrogel-type hemostatic kit

[0077] To manufacture a hydrogel-type hemostatic kit, 0.5 to 1 g of the hemostatic composition manufactured in Examples 1 to 5 was filled into a syringe, and 2 to 5 ml of a 5% calcium chloride solution was filled into the syringe.

[0078] Manufacturing Examples 42 to 54: Manufacturing of a hydrogel-type hemostatic kit

[0079] To manufacture a hydrogel-type hemostatic kit, 0.5 to 1 g of the hemostatic composition manufactured in Comparative Examples 1 to 13 was filled into a syringe, and 2 to 5 ml of a 5% calcium chloride solution was filled into the syringe.

[0080] Manufacturing Examples 55-59: Manufacturing of a hydrogel-type hemostatic kit

[0081] A hemostatic composition containing a stabilizer, a binder, thrombin, a polysaccharide, and an antibiotic having the same composition as Examples 1 to 5, except that the stabilizer, binder, and thrombin were not coated on the polysaccharide, was filled into a syringe at 0.5 to 1 g, and a 5% calcium chloride solution was filled into the syringe to prepare a hydrogel-type hemostatic kit.

[0082] Manufacturing Examples 60 to 72: Manufacturing of a hydrogel-type hemostatic kit

[0083] A hemostatic composition containing a stabilizer, a binder, thrombin, a polysaccharide, and an antibiotic having the same composition as Comparative Examples 1 to 13, except that the stabilizer, binder, and thrombin were not coated on the polysaccharide, was filled into a syringe at 0.5 to 1 g, and a 5% calcium chloride solution was filled into the syringe to prepare a hydrogel-type hemostatic kit.

[0084] Experimental Example 1: Physical Curtain Formation Test

[0085] A physical film formation test was performed to confirm whether the hemostatic composition has physical properties for a hemostatic effect by confirming the physical film formation of the hemostatic kits manufactured in Manufacturing Examples 1 to 72.

[0086] First, 1 g of the hemostatic kit manufactured in Manufacturing Examples 1 to 36 was placed on a weighing dish, and 5 ml of distilled water was dropped using a dropper. After 30 seconds, it was visually confirmed whether the hemostatic composition formed a physical film.

[0087] Next, a syringe filled with 1 g of the hemostatic composition manufactured in Manufacturing Examples 37 to 72 and a syringe filled with 5 ml of a 5% calcium chloride solution were mixed 20 times and injected onto a weighing dish, and then it was visually confirmed whether the hemostatic composition formed a physical film.

[0088] Whether a sample membrane is formedWhether a sample membrane is formedWhether a sample membrane is formedManufacturing example 1OManufacturing example 26OManufacturing example 51XManufacturing example 2OManufacturing example 27OManufacturing example 52OManufacturing example 3OManufacturing example 28OManufacturing example 53OManufacturing example 4OManufacturing example 29OManufacturing example 54OManufacturing example 5OManufacturing example 30OManufacturing example 55OManufacturing example 6OManufacturing example 31OManufacturing example 56OManufacturing example 7OManufacturing example 32OManufacturing example 57OManufacturing example 8OManufacturing example 33XManufacturing example 58OManufacturing example 9OManufacturing example 34OManufacturing example 59OManufacturing example 10OManufacturing example 35OManufacturing example 60OManufacturing example 11OManufacturing example 36OManufacturing example 61O Manufacturing Example 12O Manufacturing Example 37O Manufacturing Example 62O Manufacturing Example 13O Manufacturing Example 38O Manufacturing Example 63O Manufacturing Example 14O Manufacturing Example 39O Manufacturing Example 64O Manufacturing Example 15X Manufacturing Example 40O Manufacturing Example 65O Manufacturing Example 16O Manufacturing Example 41O Manufacturing Example 66O Manufacturing Example 17O Manufacturing Example 42O Manufacturing Example 67O Manufacturing Example 18O Manufacturing Example 43O Manufacturing Example 68O Manufacturing Example 19O Manufacturing Example 44O Manufacturing Example 69X Manufacturing Example 20O Manufacturing Example 45O Manufacturing Example 70O Manufacturing Example 21O Manufacturing Example 46O Manufacturing Example 71O Manufacturing Example 22O Manufacturing Example 47O Manufacturing Example 72O Manufacturing Example 23O Manufacturing Example Manufacturing example 48O Manufacturing example 24O Manufacturing example 49O Manufacturing example 25O Manufacturing example 50O

[0089] As shown in Table 3, in Manufacturing Examples 15, 33, 51, and 69, which contained a low amount of carboxymethyl starch due to excessive addition of stabilizer, no physical film was formed, but in the remaining Manufacturing Examples, a physical film was well formed.

[0090] Experimental Example 2: Adhesion Test

[0091] An adhesive strength test was performed to confirm whether the hemostatic composition has the effect of fixing to the bleeding site by confirming the adhesive strength of the hemostatic kits manufactured in the remaining manufacturing examples, excluding Manufacturing Examples 15, 33, 51, and 69, which failed to form a physical barrier in Experimental Example 1.

[0092] First, 1 g of the powdered hemostatic kit was placed on one end of the LDPE film, and 5 ml of distilled water was dropped using a dropper. After 5 minutes, the LDPE film was slowly tilted, and the time required for the powdered hemostatic kit to move 10 cm from its initial position was recorded.

[0093] Next, a syringe filled with 1 g of a hydrogel-type hemostatic composition and a syringe filled with 5 ml of a 5% calcium chloride solution were mixed 20 times, and 1 ml was placed at one end of an LDPE film. After 5 minutes, the LDPE film was slowly tilted, and the time required for the powder-type hemostatic kit to move 10 cm from its initial position was recorded.

[0094] Sample arrival time Sample arrival time Sample arrival time Manufacturing example 131.8±2.39 Manufacturing example 2834.8±2.59 Manufacturing example 5644.6±1.36 Manufacturing example 230.2±1.10 Manufacturing example 2926.8±1.10 Manufacturing example 5739.2±1.17 Manufacturing example 331.2±1.64 Manufacturing example 3012.4±2.41 Manufacturing example 5837.8±0.98 Manufacturing example 432.0±2.00 Manufacturing example 3111.6±1.34 Manufacturing example 5942.6±2.58 Manufacturing example 532.8±3.19 Manufacturing example 3214.0±1.22 Manufacturing example 6043.6±2.06 Manufacturing example 630.6±3.05 Manufacturing Example 3435.8±1.92 Manufacturing Example 6143.2±2.14 Manufacturing Example 732.6±3.36 Manufacturing Example 3539.6±1.95 Manufacturing Example 6229.2±1.94 Manufacturing Example 813.6±2.07 Manufacturing Example 3642.0±2.74 Manufacturing Example 6328.2±1.47 Manufacturing Example 911.8±1.48 Manufacturing Example 3745.4±3.91 Manufacturing Example 6442.4±3.67 Manufacturing Example 1028.2±1.79 Manufacturing Example 3839.6±2.30 Manufacturing Example 6541.0±1.67 Manufacturing Example 1129.2±1.48 Manufacturing Example 3941.4±5.50 Manufacturing Example 6633.0±0.89 Manufacturing Example 1216.6±1.52 Manufacturing Example 4040.8±1.30 Manufacturing Example 6730.8±1.72 Manufacturing Example 1313.4±1.52 Manufacturing Example 4143.0±2.00 Manufacturing Example 6828.4±2.50 Manufacturing Example 1412.8±2.05 Manufacturing Example 4243.0±4.30 Manufacturing Example 7043.4±2.87 Manufacturing Example 1637.6±2.70 Manufacturing Example 4343.4±0.89 Manufacturing Example 7142.0±2.61 Manufacturing Example 1729.8±1.79 Manufacturing Example 4426.6±2.61 Manufacturing Example 7244.4±1.02 Manufacturing Example 1831.0±3.08 Manufacturing Example 4527.2±2.39 Manufacturing Example 1930.0±2.83 Manufacturing Example 4645.6±1.14 Manufacturing Example 2028.6±1.67 Manufacturing Example 4740.6±1.67 Manufacturing Example 2131.8±2.39 Manufacturing Example 4830.0±2.24 Manufacturing Example 2229.8±2.17 Manufacturing Example 4928.2±2.49 Manufacturing Example 2333.2±1.64 Manufacturing Example 5024.2±4.44 Manufacturing Example 2431.4±2.70 Manufacturing Example 5243.8±2.32 Manufacturing Example 2531.6±2.61 Manufacturing Example 5341.6±1.36 Manufacturing example 2614.0±3.00 Manufacturing example 5444.0±4.Manufacturing example 15 2716.4±2.51 Manufacturing example 5541.2±1.47.

[0095] As shown in Table 4, it was confirmed that the manufacturing examples 8, 9, 12, 13, 26, 27, 30, 31, 44, 45, 48, 49, 62, 63, 66, and 67 without binder had lower adhesive strength than the manufacturing examples with binder, and it was also confirmed that the manufacturing examples 14, 32, 50, and 68 with less binder had lower adhesive strength.

[0096] Experimental Example 3: Comparison of in vitro hemostatic activity

[0097] In order to confirm the hemostatic ability in an in vitro environment of the hemostatic kits manufactured in the remaining manufacturing examples, excluding Manufacturing Examples 8, 9, 12-14, 26, 27, 30-32, 44, 45, 48-50, 62, 63, and 66-68, which had relatively insufficient adhesive strength in Experimental Example 2, a comparative clotting time test using canine whole blood was performed. Canine whole blood was administered to a test tube, and 0.025 M calcium chloride aqueous solution was added to the blood to prevent interference from anticoagulants. After administering the sample at 37°C, the test tube was tilted at 5-second intervals to check for blood clotting. If the blood did not coagulate, it was left at 37°C for 5 seconds and rechecked. The above process was repeated until the blood coagulated, and the time at which the blood coagulated was recorded and the results are presented.

[0098] Sample hemostasis time (s) Sample hemostasis time (s) Manufacturing example 112.4±0.89 Manufacturing example 387.0±0.71 Manufacturing example 212.0±0.71 Manufacturing example 396.2±0.45 Manufacturing example 311.6±1.14 Manufacturing example 406.6±0.55 Manufacturing example 412.2±0.84 Manufacturing example 417.0±1.00 Manufacturing example 511.0±0.71 Manufacturing example 427.4±0.89 Manufacturing example 612.6±0.89 Manufacturing example 436.2±0.84 Manufacturing example 711.6±1.34 Manufacturing example 46278.0±42.61 Manufacturing example 10297.0±23.55 Manufacturing example 47325.4±19.26 Manufacturing Example 11284.0±49.92 Manufacturing Example 5213.2±2.64 Manufacturing Example 1623.6±3.13 Manufacturing Example 5318.8±2.23 Manufacturing Example 1730.6±0.89 Manufacturing Example 547.4±0.80 Manufacturing Example 1810.6±0.89 Manufacturing Example 5510.6±1.36 Manufacturing Example 1917.0±1.87 Manufacturing Example 5610.8±0.75 Manufacturing Example 2017.0±1.87 Manufacturing Example 5710.2±0.40 Manufacturing Example 2118.2±0.84 Manufacturing Example 5810.0±0.89 Manufacturing Example 2217.6±1.52 Manufacturing Example 5910.4±0.49 Manufacturing Example 2318.8±1.10 Manufacturing Example 6010.2±1.60 Manufacturing Example 2420.4±1.14 Manufacturing Example 6110.0±1.10 Manufacturing Example 2515.6±0.89 Manufacturing Example 64241.8±20.92 Manufacturing Example 28279.8±17.92 Manufacturing Example 65267.0±32.34 Manufacturing Example 29314.0±36.12 Manufacturing Example 7016.2±2.04 Manufacturing Example 3428.6±0.89 Manufacturing Example 7120.2±1.94 Manufacturing Example 3533.2±1.79 Manufacturing Example 729.8±0.75 Manufacturing Example 3619.2±1.30 Manufacturing example 377.2±0.84

[0099] As shown in Table 5, it was confirmed that the manufacturing examples 10, 11, 28, 29, 46, 47, 64, and 65 that did not contain thrombin had poor hemostatic ability, and the manufacturing examples 16, 34, 52, and 70 that contained excessive binder and the manufacturing examples 17, 35, 53, and 71 that contained little thrombin had relatively poor hemostatic ability compared to the other manufacturing examples. When the binder was excessively contained, the hemostatic ability was poor because the binder prevented thrombin from coming into contact with blood. It was confirmed that the manufacturing examples 1 to 7, 18, 37 to 43, and 54 had superior hemostatic ability than the manufacturing examples 19 to 25, 36, 55 to 61, and 72 that did not have the coating due to the coating of the stabilizer, thrombin, and binder. Through Manufacturing Examples 7, 25, 43 and 61, it was confirmed that excessive addition of thrombin did not affect the improvement of hemostatic ability.

[0100] Experimental Example 4: Decomposition Test

[0101] In Experimental Example 3, except for Manufacturing Examples 10, 11, 16, 17, 19-25, 28, 29, 34-36, 46, 47, 52, 53, 55-61, 64, 65, and 70-72, which had relatively poor hemostatic ability, a degradation test was conducted to determine whether there was a risk of inflammation due to residual undegradable substances by measuring the degradation rate of the hemostatic kits manufactured in the remaining Manufacturing Examples. First, the weight of the hemostatic composition was measured. Then, each sample was placed in a conical tube, and 10 ml of 1× SBF containing 0.05 mg / ml of amylase was added. The tube containing the sample was placed in a shaking incubator at 37℃ and 80 rpm and left for 24 to 120 hours. After 24, 48, and 72 hours, the samples were taken out, filtered under reduced pressure, dried, and then weighed. The weight loss rate calculated by substituting the weight of the sample measured before and after the decomposition test into the calculation formula described below was expressed as the decomposition rate.

[0102] Calculation formula: Decomposition (%) = (W0-Wt) / W0 * 100 (%)

[0103] W0: Initial weight (g), Wt: Final weight (g)

[0104] Sample weight reduction ratio (%) 24H 48H 72H 120H Manufacturing example 192.409 2.609 3.809 9.95 Manufacturing example 292.679 3.1196.2599.00 Manufacturing example 392.5193.2295.0899.10 Manufacturing example 491.9293.0494.5199.97 Manufacturing example 592.4793.2994.6099.97 Manufacturing example 630.4130.4430.5030.66 Manufacturing example 730.0730.1230.1933.33 Manufacturing example 1891.3393.7497.3698.71 Manufacturing example Manufacturing Example 3793.2293.7995.1598.81 Manufacturing Example 3892.1793.3596.0799.92 Manufacturing Example 3993.4695.1097.7499.98 Manufacturing Example 4091.2893.3696.2298.94 Manufacturing Example 4192.4194.5597.0899.99 Manufacturing Example 4230.3430.5730.8431.02 Manufacturing Example 4329.3529.7730.0930.12 Manufacturing Example 5491.0894.5297.9199.95

[0105] As shown in Table 6, manufacturing examples 1 to 5, 18, 37 to 41 and 54 containing polysaccharides, stabilizers, binders, thrombin and antibiotics had excellent decomposition rates, but manufacturing examples 6, 7, 42 and 43 using collagen or gelatin instead of polysaccharides did not decompose properly even after 120 hours.

[0106] Experimental Example 5: Changes in thrombin activity following long-term storage

[0107] In order to confirm whether the hemostatic compositions of Preparation Examples 1, 18, 37, and 54 according to the present invention maintain the activity of thrombin when stored for a long period of time at room temperature, the change in thrombin activity according to the accelerated aging period was measured. When the hemostatic compositions were collected at a specific time (a period corresponding to real-time) under accelerated aging conditions and reacted with fibrinogen, the time for fibrin formation was measured, and if the activity was maintained at 80% or more of the initial level, it was determined to be effective, and the results were presented.

[0108] Storage period (month) 0121824 Thrombin activity Manufacturing example 1100% 96.3% 91.4% 89.6% Manufacturing example 18100% 80.7% 73.4% 67.8% Manufacturing example 37100% 98.5% 93.8% 92.6% Manufacturing example 54100% 84.2% 80.1% 78.9%

[0109] As shown in Table 7, it was confirmed that manufacturing examples 1 and 37, which contained an appropriate amount of stabilizer, maintained thrombin activity well for 24 months, while manufacturing examples 18 and 54, which contained a small amount of stabilizer, showed a significant decrease in thrombin activity.

[0110] Experimental Example 6: Changes in antibacterial activity according to antibiotic content

[0111] In order to confirm the antibacterial activity of the hemostatic compositions of Preparation Examples 1, 2, 12, 18, 37, 38, 48 and 54 according to the present invention, the degree of bacterial growth against Staphylococcus aureus was confirmed. LB Agar medium was prepared, 9 log CFU / ml of Staphylococcus aureus was smeared, and 1 g of the hemostatic composition was placed in the center of the medium. After culturing for 24 hours in a 37°C incubator, it was visually confirmed whether bacterial growth occurred around the hemostatic composition, and when there was no bacterial growth around the composition, it was determined to have antibacterial activity, and the results were presented.

[0112] Sample Bacteria Inhibition Antibacterial Power Manufacturing Example 10 Manufacturing Example 2 Manufacturing Example 12 Manufacturing Example 18X Manufacturing Example 37 Manufacturing Example 38 Manufacturing Example 48 Manufacturing Example 54X

[0113] As shown in Table 8, Preparation Examples 1, 2, 37, and 38, which contained an appropriate amount of antibiotics, successfully inhibited bacterial growth, while Preparation Examples 18 and 54, which contained a small amount of antibiotics, did not effectively inhibit bacterial growth. In addition, Preparation Examples 12 and 48, which contained an excessive amount of antibiotics, were confirmed to successfully inhibit bacterial growth, similar to Preparation Examples 1 and 2.

[0114] Experimental Example 7: Comparison of Cytotoxicity in Fibroblasts

[0115] To confirm the safety of the powdered hemostatic compositions prepared in Manufacturing Examples 1, 2, 12, and 18 and the hydrogel-type hemostatic kits prepared in Manufacturing Examples 37, 38, 48, and 54, a cytotoxicity comparison test was performed on fibroblasts. After culturing fibroblasts, a solution obtained by dissolving the hemostatic composition in cell media was added to the cell culture medium and cultured for 24 hours. The number of surviving cells after culture was measured using the MTT assay, and the results are presented.

[0116] Cell viability comparison group 100% manufacturing example 1100% manufacturing example 2106% manufacturing example 1272% manufacturing example 18101% manufacturing example 37115% manufacturing example 38102% manufacturing example 4863% manufacturing example 54103%

[0117] * Comparison group: Untreated group

[0118] As shown in Table 7, Preparation Examples 1, 2, 37, and 38, which contained an appropriate amount of antibiotics, showed no cytotoxicity with cell viability of 100% or more, but Preparation Examples 12 and 48, which contained an excessive amount of antibiotics, showed toxicity with reduced cell viability. In addition, Preparation Examples 18 and 54, which contained a small amount of antibiotics, were confirmed to be non-cytotoxic, similar to Preparation Examples 1, 2, 37, and 38.

[0119] Experimental Example 8: Comparison of hemostatic activity in a rat liver injury model.

[0120] To confirm the hemostatic ability of the powder-type hemostatic kit manufactured in Manufacturing Example 1 and the hydrogel-type hemostatic kit manufactured in Manufacturing Example 37, a hemostatic ability comparison test was performed in a rat liver injury model. After inducing bleeding by punching the liver, gauze was applied for 30 seconds, the composition was applied, and pressure was applied for 15 seconds, and the presence or absence of bleeding was checked. If bleeding persisted, hemostatic pressure was applied for an additional 15 seconds, and this was repeated until hemostasis was complete. The time for successful hemostasis was measured and the results are presented.

[0121] Hemostasis time (s) Comparison group 240.63 ± 26.98 s Manufacturing example 141.25 ± 9.92 s Manufacturing example 3770 ± 29.05 s

[0122] * Control group: Gauze treatment alone

[0123] Experimental Example 9: Comparison of hemostatic activity in a porcine gastrointestinal injury model.

[0124] In order to confirm the hemostatic ability of the endoscopic powder hemostatic kit including the powder hemostatic kit manufactured in Manufacturing Example 1 and the endoscopic spray device, a hemostatic ability comparison test was performed in a porcine gastrointestinal injury model. After inducing bleeding classified as Forrest lb through gastric mucosal resection using an endoscope, the composition was applied using the endoscopic spray device, and the presence or absence of bleeding was checked after 2 minutes. If bleeding persisted, the composition was additionally applied, and the presence or absence of bleeding was rechecked after 1 minute, and this was repeated until hemostasis was complete. The time for successful hemostasis was measured and the results are presented.

[0125] Hemostasis time (s) Comparison group 434.20 ± 24.67 s Manufacturing example 1140.20 ± 11.20 s

[0126] * Control group: No hemostatic agent treatment

[0127] Experimental Example 10: Microstructure Confirmation Using a Scanning Electron Microscope (SEM)

[0128] To confirm whether the hemostatic composition according to the present invention forms a hydrogel when mixed with a cationic solution, the microstructure was observed using SEM. The hemostatic composition before mixing (Example 1) and the hemostatic composition (Example 1) mixed with a cationic (calcium chloride) solution (Preparation Example 37) and then freeze-dried were observed using SEM to obtain microstructural images.

[0129] As a result, it was confirmed that the hemostatic composition in powder form was converted into a hydrogel by forming a porous mesh when mixed with a cationic solution (Fig. 4).

[0130] While specific aspects of the present invention have been described in detail above, it will be apparent to those skilled in the art that these specific descriptions merely represent preferred embodiments and are not intended to limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A hemostatic composition comprising (a) a polysaccharide coated with a mixture of a stabilizer, a binder and thrombin; and (b) an antibiotic.

2. A hemostatic composition according to claim 1, characterized in that the stabilizer is at least one selected from the group consisting of serum albumin, mannitol, sorbitol, sucrose, trehalose, dextran, glucose, sodium acetate, and glycine.

3. A hemostatic composition according to claim 2, characterized in that the stabilizer is mannitol or sorbitol or a mixture of the two.

4. A hemostatic composition according to claim 1, characterized in that the binder is at least one selected from the group consisting of polypyrrolidone (PVP), polyethylene glycol (PEG), and polyethylene oxide (PEO).

5. A hemostatic composition according to claim 4, characterized in that the binder is polypyrrolidone.

6. A hemostatic composition according to claim 1, characterized in that the polysaccharide is at least one selected from the group consisting of starch, alginate, pullulan, carboxymethyl cellulose, and dextrin.

7. A hemostatic composition according to claim 6, characterized in that the polysaccharide is starch.

8. A hemostatic composition according to claim 6, wherein the starch is carboxymethyl starch and has a molecular weight of 500,000 to 1,000,000 g / mol.

9. A hemostatic composition according to claim 1, characterized in that the antibiotic is at least one selected from the group consisting of gentamicin, penicillin, cephalosporin, monobactam, and carbapenem.

10. A hemostatic composition according to claim 9, characterized in that the antibiotic is gentamicin.

11. A hemostatic composition characterized in that, in the first paragraph, the polysaccharide coated with a mixture of a stabilizer, a binder, and thrombin comprises 1 to 45 wt% of the stabilizer, 2 to 10 wt% of the binder, 0.01 to 10 wt% of the thrombin, and 35 to 96.99 wt% of the polysaccharide, and the antibiotic is contained in an amount of 0.5 to 1.5 wt% based on 100 wt% of the polysaccharide coated with the mixture of a stabilizer, a binder, and thrombin.

12. In the first paragraph, the polysaccharide coated with a mixture of a stabilizer, a binder and thrombin is (a) a step of dissolving a stabilizer, a binder and thrombin in distilled water and homogenizing the mixture; (b) a step of immersing the polysaccharide in a homogenizing solution; (c) a step of separating the immersed polysaccharide from the solution; and (d) A hemostatic composition characterized by being manufactured by a method including the step of freeze-drying and pulverizing the separated polysaccharide. 13.(a) A hemostatic composition comprising a polysaccharide coated with a mixture of a stabilizer, a binder and thrombin and (b) an antibiotic; and a powder-type hemostatic kit comprising a cationic substance.

14. A powder-type hemostatic kit according to claim 13, characterized in that the cationic substance is at least one selected from the group consisting of calcium chloride (CaCl2) and sodium chloride (NaCl).

15. A powder-type hemostatic kit, characterized in that the cationic substance is contained in an amount of 0.01 to 5 parts by weight based on 100 parts by weight of the hemostatic composition in the 14th paragraph.

16. An endoscopic powder hemostatic kit comprising the powder hemostatic kit of Article 13 and an endoscopic spray device. 17.(a) a first container comprising a hemostatic composition comprising a polysaccharide coated with a mixture of a stabilizer, a binder and thrombin and (b) an antibiotic; a second container containing a cationic solution; and A hydrogel-type hemostatic kit comprising a connector connecting a first container and a second container.

18. A hydrogel-type hemostatic kit according to claim 17, characterized in that the cationic solution is at least one selected from the group consisting of purified water, calcium chloride (CaCl2) solution, and sodium chloride (NaCl) solution.

19. A hydrogel-type hemostatic kit according to claim 17, characterized in that the cationic solution is prepared by adding a water-soluble solvent to 0.01 to 5 parts by weight of a cationic substance per 100 parts by weight of a hemostatic composition based on dry powder.

20. A hydrogel-type hemostatic kit, characterized in that, in the 17th paragraph, when the hemostatic composition and the cationic solution are mixed, a hydrogel is formed, and the average pore size of the hydrogel is 100 μm to 1000 μm.

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