Hemostatic composition

Abstract

The present disclosure relates to hemostatic compositions comprising a biocompatible polymer, thrombin, and hemofibrinogen, and methods for preparing such compositions.

Description

Technical Field

[0001] The present disclosure relates to hemostatic compositions comprising a biocompatible polymer, thrombin, and fibrinogen, and methods for preparing such compositions.

[0002] Background

[0003] Protein-based hemostatic materials, such as gelatin, are commercially available in solid sponge and loose or uncompressed powder forms for use in surgical procedures. Depending on the mixing conditions and the relative ratios of the materials, mixing a loose or uncompressed powder with a fluid, such as saline or a thrombin solution, can form a paste or slurry that can be used as a hemostatic composition for use in cases of diffuse bleeding, particularly from uneven surfaces or hard-to-reach areas.

[0004] Hemostatic pastes are typically prepared at the time of use by mechanically agitating and mixing particulate, crosslinked gelatin with a liquid, such as a thrombin solution, to provide uniformity of the composition. Mixing to form a paste typically requires thorough mixing, such as kneading or transferring between two syringes.

[0005] Hemostatic Matrix (Ethicon) is a commercially available kit for producing a hemostatic gelatin paste containing thrombin, which is prepared by transferring a gelatin matrix-thrombin solution mixture back and forth between two connected syringes. Hemostatic Matrix (Baxter) is likewise a kit for producing a hemostatic gelatin paste. Once a substantially homogeneous paste composition is obtained, the hemostatic paste can be applied to the bleeding site by extruding the paste from a syringe to promote hemostasis.

[0006] Thrombin is a well-known hemostatic adjuvant in hemostatic compositions, which acts as a serine protease that converts soluble fibrinogen into insoluble fibrin strands and catalyzes many other coagulation-related reactions. The combination of thrombin and fibrinogen is also used in certain currently clinically used hemostatic products, such as sponges, patches, glues, and sealants containing thrombin and fibrinogen. However, since thrombin acts on fibrinogen in the presence of water, thrombin and fibrinogen are provided separately or together in a dry form to prevent premature action of thrombin on fibrinogen. For administration, the thrombin and fibrinogen components can be administered to the patient in a dry form or mixed with an aqueous medium at the time of administration to the patient.

[0007] Although currently available hemostatic products can effectively control mild and moderate bleeding, it would be beneficial to have hemostatic compositions that can control such bleeding even faster and more effectively and that can be used to control more severe bleeding. It would be even more beneficial to have hemostatic products that can effectively control bleeding in minimally invasive surgical procedures where sponges, patches, glues, and sealants cannot be used or are less efficient.

[0008] Overview

[0009] The present disclosure provides compositions having improved hemostatic properties and methods for preparing such hemostatic compositions. The improved hemostatic properties are attributed to an optimized combination of components, resulting in improved adhesion properties. Such compositions are highly valuable in operating rooms where bleeding must be controlled in a rapid and effective manner.

[0010] Surprisingly, the inventors have demonstrated that the hemostatic paste compositions disclosed herein, which comprise a biocompatible polymer, thrombin, and fibrinogen, remain flowable for more than 1.5 hours after reconstitution. Accordingly, the paste compositions of the present disclosure can be pre-prepared and used over an extended period of time, which is highly advantageous in operating rooms and advanced surgical procedures.

[0011] Thus, in one aspect, the present disclosure provides a hemostatic composition comprising:

[0012] a) one or more biocompatible polymers in particulate form,

[0013] b) fibrinogen in an amount of 1 to 150 mg per gram of biocompatible polymer, and

[0014] c) thrombin in an amount of 100 to 5000 IU per gram of biocompatible polymer.

[0015] In a second aspect, the present disclosure relates to a method for preparing a hemostatic composition, the method comprising the steps of:

[0016] a) providing a dry hemostatic composition as described herein, and

[0017] b) adding an amount of an aqueous medium to the dry hemostatic composition of a).

[0018] The amount of aqueous medium added is generally an amount sufficient to obtain a hemostatic composition in paste form.

[0019] In another aspect, the present disclosure provides a hemostatic composition comprising:

[0020] a) one or more biocompatible polymers in particulate form in an amount of 7% to 34% by weight,

[0021] b) fibrinogen in an amount of 0.008% to 5% by weight,

[0022] c) thrombin in an amount of 7 to 1700 IU per gram, and

[0023] d) an aqueous medium.

[0024] Hemostatic compositions are typically provided in a paste form, i.e., an aqueous medium is present in an amount sufficient to provide the composition in a paste form.

[0025] In yet another aspect, the present disclosure relates to the hemostatic compositions described herein for promoting hemostasis and / or wound, bone, tendon, and / or tissue healing in an individual in need thereof.

[0026] In another aspect, the present disclosure relates to a container comprising the hemostatic composition described herein.

[0027] In a last aspect, the present disclosure provides a kit comprising:

[0028] a) a first container containing the hemostatic composition as described herein,

[0029] b) a second container containing an aqueous medium; and

[0030] c) an optional outer package,

[0031] wherein the two containers are interconnected.

[0032] Definitions

[0033] As used herein, the term "about" referring to an amount or percentage shall be construed as varying by ±10%, such as ±5%, relative to the value of the amount or percentage to which it refers.

[0034] "Bioactive agent" is any agent, drug, compound, substance composition, or mixture that provides some pharmacological (usually beneficial) effect demonstrable in vivo or in vitro. An agent is considered bioactive if it interacts with or has an effect on the cellular tissue in a human or animal body. As used herein, this term further includes any physiological or pharmacological active substance that produces a local or systemic effect in an individual. A bioactive agent can be a protein, such as an enzyme. Other examples of bioactive agents include, but are not limited to, agents comprising or consisting of: oligosaccharides, polysaccharides, optionally glycosylated peptides, optionally glycosylated polypeptides, oligonucleotides, polynucleotides, lipids, fatty acids, fatty acid esters, and secondary metabolites. It can be used prophylactically and therapeutically in combination with the treatment of an individual, such as a human or any other animal. The term "bioactive agent" as used herein does not cover cells, such as eukaryotic or prokaryotic cells.

[0035] "Biocompatibility" refers to the ability of a material to perform its intended function without inducing any substantial undesirable local or systemic effects in a host.

[0036] "Biologically absorbable" or "resorbable" is the term used in the context of the present text to describe a material from which the powder is made that can degrade in the body into smaller molecules that have dimensions that permit their transport into the bloodstream. Through such degradation and absorption, the powder material will gradually be removed from the site of application. For example, gelatin can be degraded by proteolytic tissue enzymes into absorbable smaller molecules, and thus, gelatin is generally absorbed within about 4 - 6 weeks when applied to tissue and within 3 - 5 days when applied to bleeding surfaces and mucous membranes.

[0037] "Hemostasis" is the process of reducing or stopping bleeding. Hemostasis occurs when blood is present outside the body or a blood vessel and is the body's instinctive response to stop bleeding and blood loss. During hemostasis, three steps occur rapidly in sequence. Vasospasm is the first response as the blood vessels constrict to reduce blood loss. In the second step, a platelet plug forms as platelets adhere together to form a temporary seal over the rupture in the blood vessel wall. The third and final step is called coagulation or blood clotting. Coagulation uses fibrin threads that act as "molecular glue" to reinforce the platelet plug. Thus, a hemostatic compound is capable of stimulating hemostasis.

[0038] "International Unit (IU)". In pharmacology, the international unit is a unit of measure of the amount of a substance based on its biological activity or effect. Its abbreviations are IU, UI, or IE. It is used to quantify vitamins, hormones, some drugs, vaccines, blood products, and similar biologically active substances.

[0039] A "paste" according to the present disclosure has a malleable, putty-like consistency, such as toothpaste. A paste is a thick fluid mixture of a pulverized solid / a solid in powder form and a liquid. A paste is a material that behaves like a solid until a sufficiently large load or stress is applied, at which point it flows like a fluid, i.e., a paste is flowable. A flowable material conforms effectively to an irregular surface upon application. A paste typically consists of a suspension of particulate material in a background fluid. The individual particles are packed together like sand on a beach, forming a disordered, glassy, or amorphous structure and imparting solid-like characteristics to the paste. It is this "packing together" that gives the paste some of its most unusual properties; this makes the paste exhibit the characteristics of a brittle material. A paste is not a gel / jelly. A "slurry" is a fluid mixture of a powdered / crushed solid and a liquid (such as water). A slurry behaves like a thick fluid in some respects, flows under gravity, and can be pumped if not too thick. A slurry can be functionally regarded as a thin, watery paste, but a slurry generally contains more water than a paste. Substantially water-insoluble powder particles (such as cross-linked gelatin particles) will form a paste or a slurry when mixed with an aqueous medium.

[0040] "Percentage". Unless otherwise indicated, percentages are by weight: % w / w or wt%. Ratios are indicated as weight ratios (w / w).

[0041] Detailed Description

[0042] The present disclosure provides a composition having improved hemostatic properties and a method for preparing the hemostatic composition.

[0043] Thus, in one aspect, the present disclosure provides a hemostatic composition comprising:

[0044] a) one or more biocompatible polymers in particulate form,

[0045] b) fibrinogen in an amount of 1 to 150 mg per gram of biocompatible polymer, and

[0046] c) thrombin in an amount of 100 to 5000 IU per gram of biocompatible polymer.

[0047] In a second aspect, the present disclosure relates to a method for preparing a hemostatic composition, which comprises the steps of:

[0048] a) providing a dry hemostatic composition as described herein, and

[0049] b) adding a quantity of aqueous medium to the dry hemostatic composition of a).

[0050] The advantages of the hemostatic composition disclosed herein are numerous and include:

[0051] · Improved hemostatic efficacy, for example, more severe bleeding can be stopped in a shorter time.

[0052] · Less time is spent in preparing the hemostatic composition, for example, bleeding can be stopped more quickly.

[0053] · Since there are fewer processing steps, the risk of compromising the sterility of the hemostatic composition during preparation is reduced.

[0054] · Since the preparation of the paste is simplified, the risk of errors during preparation is reduced.

[0055] · Reliably and consistently reconstituted within a short period of time.

[0056] · Superior consistency and adhesion properties reduce the need for pressure.

[0057] · Well-suited for minimally invasive surgery (MIS), including robotic surgery.

[0058] · Can be applied to sprayable patches.

[0059] · Avoids the time-consuming and error-prone dilution step of standard hemostatic composition products.

[0060] · Minimizes operating room costs because the preparation of the currently described product is so simple and rapid that there is no reason to pre-prepare hemostatic fluids that need to be discarded before surgery.

[0061] Dry hemostatic composition

[0062] The present invention relates to a hemostatic composition comprising:

[0063] a) one or more biocompatible polymers in particulate form,

[0064] b) fibrinogen, and

[0065] c) thrombin.

[0066] In one embodiment, the present disclosure relates to a hemostatic composition comprising:

[0067] a) one or more biocompatible polymers in particulate form,

[0068] b) fibrinogen in an amount of 1 to 150 mg per gram of biocompatible polymer, and

[0069] c) thrombin in an amount of 100 to 5000 IU per gram of biocompatible polymer.

[0070] The hemostatic composition is typically provided in a substantially dry, storage-stable form. In one embodiment, the dry composition is storage-stable at room temperature for at least 12 months, preferably at least 24 months.

[0071] In one embodiment, the hemostatic composition comprises one or more biocompatible polymers in particulate form in an amount of at least 80% by weight of the composition, such as at least 81% by weight of the composition, such as at least 83% by weight of the composition, such as at least 85% by weight of the composition, such as at least 87% by weight of the composition, such as at least 90% by weight of the composition, such as at least 91% by weight of the composition, such as at least 95% by weight of the composition.

[0072] In one embodiment, the hemostatic composition comprises one or more biocompatible polymers in particulate form in an amount of 80% to 99% by weight of the composition, such as 81% to 99% by weight of the composition, such as 82% to 99% by weight of the composition, such as 83% to 99% by weight of the composition, such as 84% to 99% by weight of the composition, such as 85% to 99% by weight of the composition, such as 86% to 99% by weight of the composition, such as 87% to 99% by weight of the composition, such as 88% to 99% by weight of the composition, such as 89% to 99% by weight of the composition, such as 90% to 99% by weight of the composition.

[0073] In one embodiment, the hemostatic composition comprises one or more biocompatible polymers in particulate form, in an amount of 85% to 99% by weight of the composition, such as 85% to 98% by weight of the composition, such as 85% to 97%, such as 85% to 96%, such as 85% to 96%, such as 85% to 95%.

[0074] For example, in one embodiment, the hemostatic composition comprises one or more biocompatible polymers in particulate form, in an amount of 83% to 97% by weight of the composition, such as 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96% or 97% by weight of the composition.

[0075] In one embodiment, the hemostatic composition comprises one or more biocompatible polymers in particulate form in an amount of 85% to 97% by weight of the composition. In one embodiment, the hemostatic composition comprises one or more biocompatible polymers in particulate form in an amount of 89% to 97% by weight of the composition. In one embodiment, the hemostatic composition comprises one or more biocompatible polymers in particulate form in an amount of 90% to 97% by weight of the composition. In one embodiment, the hemostatic composition comprises one or more biocompatible polymers in particulate form in an amount of 92% to 97% by weight of the composition. In one embodiment, the hemostatic composition comprises one or more biocompatible polymers in particulate form in an amount of 92%, 93%, 94%, 95%, 96% or 97% by weight of the composition.

[0076] In one embodiment, the present disclosure relates to a hemostatic composition comprising:

[0077] a) one or more biocompatible polymers in particulate form,

[0078] b) fibrinogen, in an amount of 1 to 150 mg per gram of biocompatible polymer, and

[0079] c) thrombin, in an amount of 100 to 5000 IU per gram of biocompatible polymer;

[0080] wherein one or more biocompatible polymers in particulate form are present in an amount of at least 80% by weight of the composition, such as at least 81% by weight of the composition, such as at least 83% by weight, such as at least 85% by weight, such as at least 87% by weight, such as at least 90% by weight, such as at least 91% by weight, such as at least 95% by weight.

[0081] In one embodiment, the present disclosure relates to a hemostatic composition comprising:

[0082] a) one or more biocompatible polymers in particulate form,

[0083] b) fibrinogen in an amount of 1 to 150 mg per gram of biocompatible polymer, and

[0084] c) thrombin in an amount of 100 to 5000 IU per gram of biocompatible polymer;

[0085] wherein one or more biocompatible polymers in particulate form are present in an amount of 85% to 97% by weight of the composition, such as 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96% or 97% by weight of the composition.

[0086] In one embodiment, the present disclosure relates to a hemostatic composition comprising:

[0087] a) one or more biocompatible polymers in particulate form,

[0088] b) fibrinogen in an amount of 1 to 150 mg per gram of biocompatible polymer, and

[0089] c) thrombin in an amount of 100 to 5000 IU per gram of biocompatible polymer;

[0090] wherein one or more biocompatible polymers in particulate form are present in an amount of 90% to 97% by weight of the composition.

[0091] In one embodiment, the hemostatic composition contains less than 10% water by weight, preferably less than 5% water by weight, preferably less than 1% water by weight.

[0092] Biocompatible polymer

[0093] The biocompatible polymers of the present disclosure can be biological or non-biological polymers. Suitable biological polymers include proteins such as gelatin, collagen, albumin, hemoglobin, casein, fibrinogen, fibrin, fibronectin, elastin, keratin, and laminin; or derivatives or combinations thereof. Particularly preferred is the use of gelatin or collagen, and more preferably gelatin. Other suitable biological polymers include polysaccharides such as glycosaminoglycans, starch derivatives, xylan, cellulose derivatives, hemicellulose derivatives, agarose, alginate, and chitosan; or derivatives or combinations thereof. Suitable non-biological polymers will be selected to be degradable by either of two mechanisms, namely (1) polymer backbone decomposition or (2) generation of water-soluble side chain degradation. Exemplary non-biological polymers include synthetic materials such as polyacrylates, polymethacrylates, polyacrylamides, polyethylene resins, polylactide-glycolide, polycaprolactone, and polyethylene oxide; or derivatives or combinations thereof. Combinations of different types of polymers are also possible.

[0094] In one embodiment, the biocompatible polymer in particulate form comprises a biocompatible polymer selected from the group consisting of or consisting of a biocompatible polymer selected from the group consisting of: gelatin, collagen, chitin, chitosan, alginate, cellulose, oxidized cellulose, carboxymethyl cellulose, polyglycolic acid, polyacetic acid, and combinations thereof.

[0095] In one embodiment, the biocompatible polymer comprises or consists of powder particles that are substantially insoluble in an aqueous medium.

[0096] In one embodiment, the biocompatible polymer is bioabsorbable. Examples of suitable bioabsorbable materials include gelatin, collagen, chitin, chitosan, alginate, cellulose, oxidized cellulose, polyglycolic acid, polyacetic acid, and combinations thereof. It should be understood that the present disclosure also encompasses various forms thereof, such as linear or crosslinked forms, salts, esters, and the like. In a preferred embodiment of the present invention, the bioabsorbable material comprises or consists of gelatin. Gelatin is preferred because gelatin has a high degree of bioabsorbability. In addition, gelatin has a high degree of biocompatibility, meaning that it is non-toxic to animals (such as humans) when entering the bloodstream or upon long-term contact with human tissues.

[0097] Gelatin is typically derived from porcine sources, but can be derived from other animal sources such as bovine or fish sources. Gelatin can also be prepared synthetically, i.e., by recombinant means.

[0098] In a preferred embodiment, the biocompatible polymer is crosslinked. Crosslinking generally renders the polymer substantially insoluble in aqueous media. In one embodiment, the biocompatible polymer consists of powder particles that are substantially insoluble in aqueous media. Any suitable crosslinking method known to those skilled in the art can be used, including chemical crosslinking methods and physical crosslinking methods.

[0099] In one embodiment of the present disclosure, the polymer has been physically crosslinked, such as by dry heat crosslinking. The dry heat treatment is typically carried out at a temperature of 100 °C to 250 °C, such as about 110 °C to about 200 °C. In particular, the temperature can be in the range of 110 °C to 160 °C, for example in the range of 110 °C to 140 °C, or in the range of 120 °C to 180 °C, or in the range of 130 °C to 170 °C, or in the range of 130 °C to 160 °C, or in the range of 120 °C to 150 °C. The crosslinking period can be optimized by those skilled in the art and is typically a period of about 10 minutes to about 12 hours, such as about 1 hour to about 10 hours, for example about 2 hours to about 10 hours, such as about 4 hours to about 8 hours, for example about 5 hours to about 7 hours, such as about 6 hours.

[0100] In another embodiment, the polymer has been chemically crosslinked, i.e., by exposure to a chemical crosslinking agent. Examples of suitable chemical crosslinking agents include, but are not limited to, aldehydes (especially glutaraldehyde and formaldehyde), acyl azides, carbodiimides, hexamethylene diisocyanate, polyether oxides, 1,4-butanediol diglycidyl ether, tannic acid, aldoses (such as D-fructose), genipin, and dye-mediated photooxidation. Specific compounds include, but are not limited to, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and dithiobis(propionyl hydrazide) (DTP).

[0101] In one embodiment, the biocompatible polymer particles according to the present disclosure are obtained from crosslinked sponges such as gelatin or collagen, especially crosslinked sponges of gelatin (such as commercially available sponges and sponges). The crosslinked sponges are micronized by methods known in the art to obtain the crosslinked biocompatible polymer in powder form, such as by a rotary bed, extrusion, granulation, and treatment in a high-intensity mixer, or grinding (e.g., by using a hammer mill or a centrifugal mill).

[0102] The available from Ethicon is a gelatin-based crosslinked absorbable hemostatic sponge. It absorbs >35 g of blood / g and is completely absorbed by the human body within 4 - 6 weeks.

[0103] In one embodiment, the biocompatible polymer in particulate form comprises or consists of cross-linked gelatin particles.

[0104] In one embodiment, the cross-linked gelatin particles are obtained from micronized porous gelatin sponges or dried hydrogels. In one embodiment, the porous gelatin sponge or dried hydrogel has been cross-linked by dry heat treatment.

[0105] The porous gelatin sponge can be prepared as follows: A quantity of soluble gelatin is mixed with an aqueous medium to produce a foam containing a discontinuous gas phase, the foam is dried and the dried foam is cross-linked by exposure to dry heat. The resulting cross-linked sponge can be micronized by methods known in the art. The gelatin foam typically has a gelatin concentration of about 1% to 70% by weight, usually 3% to 20% by weight. Drying is typically carried out at about 20°C to about 40°C for about 5 to 20 hours. The dried foam is typically cross-linked by exposure to a temperature of about 110°C to about 200°C for about 15 minutes to about 8 hours, such as exposure to about 150°C to about 170°C for about 5 to 7 hours. The cross-linking period can be optimized by those skilled in the art and is typically a period of about 10 minutes to about 12 hours, such as about 1 hour to about 10 hours, for example about 2 hours to about 10 hours, such as about 4 hours to about 8 hours, for example about 5 hours to about 7 hours, such as about 6 hours.

[0106] Drying of the foam or hydrogel can also be achieved by freeze-drying by methods known to those skilled in the art.

[0107] In one embodiment, the cross-linked gelatin particles are obtained by micronizing a cross-linked porous gelatin sponge or a substantially non-porous cross-linked dried hydrogel. The gelatin can be cross-linked, for example, by exposure to glutaraldehyde (e.g., 0.01% to 0.05% w / w, overnight in an aqueous buffer at 0°C to 15°C), sodium periodate (e.g., 0.05 M, maintained at 0°C to 15°C for 48 hours), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (e.g., 0.5% to 1.5% w / w, overnight at room temperature) or by exposure to about 0.3 to 3 Mrad of gamma or electron beam radiation. When cross-linked with glutaraldehyde, the cross-linking occurs via the formation of a Schiff base that can be stabilized by subsequent reduction (e.g., by treatment with sodium borohydride).

[0108] The particles can be obtained from the dried product by methods known to those skilled in the art. In one embodiment, the gelatin particles are obtained by micronization of the dried product, such as by granulation and treatment in a high-shear mixer, grinding, for example by using a hammer mill, ball mill or centrifugal mill. In another embodiment, the particles are obtained by grinding the dried product to an appropriate size. This can be carried out, for example, by mortar and pestle, crushing and any other available physical procedures.

[0109] In one embodiment, the particles are obtained by hammer milling a sponge or a dried hydrogel. Preferably, the hammer mill has a built-in sieve to produce the desired particle size distribution.

[0110] In one embodiment of the present invention, the gelatin particles have a diameter of about 1 μm to 1000 μm, such as about 10 μm to 800 μm, for example about 50 μm to 600 μm, such as about 100 μm to 500 μm, for example about 200 μm to 500 μm, such as about 450 μm.

[0111] In one embodiment, the particle size is less than about 1000 microns such that it is capable of passing through a 1×1 mm sieve.

[0112] Generally, at least 90% of the powder particles have a size of 1 μm to 1200 μm.

[0113] In another embodiment, the average particle size of the dried particles is 1 μm to 1000 μm, such as about 10 μm to 800 μm, for example about 50 μm to 600 μm, such as about 100 μm to 500 μm, for example about 200 μm to 500 μm, such as about 450 μm.

[0114] The average particle size of the dried particles can be measured, for example, by laser diffraction.

[0115] In one embodiment, the biocompatible polymer in particulate form is present in an amount of about 0.05 to 20 g, such as about 0.2 to 10 g, such as about 0.5 to 2 g, such as about 1 g.

[0116] In one embodiment, the biocompatible polymer in particulate form is present in an amount of about 1 g.

[0117] The biocompatible polymer in particulate form used in the present disclosure is generally provided in a sterile form.

[0118] Fibrinogen

[0119] Fibrinogen or factor I is a glycoprotein complex that circulates in vertebrate blood. During tissue and vascular injury, it is enzymatically converted to fibrin by thrombin and participates in blood clot formation.

[0120] In one embodiment, the fibrinogen is human fibrinogen.

[0121] In one embodiment, the fibrinogen is recombinant human fibrinogen.

[0122] In other embodiments, the fibrinogen is derived from a mammal other than human, such as bovine fibrinogen.

[0123] In one embodiment, the fibrinogen is a dried fibrinogen composition. For example, the fibrinogen can be in the form of granules or powder. The dried fibrinogen composition can be prepared by any method known to those skilled in the art and is typically provided in a sterile form. Thus, in one embodiment, the dried fibrinogen composition is sterile.

[0124] The fibrinogen can also be coated on a biocompatible polymer in the form of granules. The coating of fibrinogen on the biocompatible polymer can be obtained by any method known in the art, such as by spray techniques, which can be carried out in any spray device. For example, a well-known method of coating granules by spraying is the fluidized bed process. Thus, in one embodiment, the fibrinogen is coated on the biocompatible polymer granules to obtain a fibrinogen layer on the biocompatible polymer granules. In one embodiment, the fibrinogen is sprayed onto the biocompatible polymer granules, for example, by fluidized bed process technology.

[0125] In one embodiment, the hemostatic composition comprises fibrinogen in an amount of about 1 to about 150 mg per gram of biocompatible polymer, such as about 5 to about 150 mg of fibrinogen per gram of biocompatible polymer, such as about 10 to about 150 mg, such as about 15 to about 150 mg, such as about 20 to about 150 mg, such as about 25 to about 150 mg, such as about 30 to about 150 mg, such as about 30 to about 125 mg, such as about 30 to about 100 mg of fibrinogen.

[0126] In one embodiment, the hemostatic composition comprises fibrinogen in an amount of about 20 to about 80 mg of fibrinogen per gram of biocompatible polymer, such as about 30 mg per gram of biocompatible polymer, such as about 35 mg, such as about 40 mg, such as about 45 mg, such as about 50 mg, such as about 55 mg, such as about 60 mg, such as about 70 mg, such as about 75 mg of fibrinogen. In one embodiment, the composition comprises about 35 mg of fibrinogen per gram of biocompatible polymer. In one embodiment, the composition comprises about 70 mg of fibrinogen per gram of biocompatible polymer.

[0127] In one embodiment, the hemostatic composition comprises fibrinogen in an amount of from about 20 mg to about 100 mg of fibrinogen per gram of biocompatible polymer, such as from about 25 mg to about 100 mg of fibrinogen per gram of biocompatible polymer, such as from about 30 mg to about 100 mg, such as 30 mg to about 95 mg, such as 30 mg to about 90 mg, such as 30 mg to about 85 mg of fibrinogen.

[0128] In one embodiment, the hemostatic composition comprises no more than 100 mg of fibrinogen per gram of biocompatible polymer. In one embodiment, the composition comprises more than 10 mg of fibrinogen per gram of biocompatible polymer.

[0129] In one embodiment, the hemostatic composition comprises from about 20 to about 100 mg of fibrinogen per gram of biocompatible polymer, such as from about 20 to about 30 mg of fibrinogen per gram of biocompatible polymer, such as from about 30 mg to about 40 mg, such as from about 40 mg to about 50 mg, such as from about 50 mg to about 60 mg, such as from about 60 mg to about 70 mg, such as from about 70 mg to about 80 mg, such as from about 80 mg to about 90 mg, such as from about 90 mg to about 100 mg of fibrinogen.

[0130] In one embodiment, when in dry form, i.e., before mixing with an aqueous medium, the hemostatic composition comprises from 1% to 15% by weight, such as from 1% to 10% by weight, such as 1% to 2%, such as 2% to 3%, such as 3% to 4%, such as 4% to 5%, such as 5% to 6%, such as 6% to 7%, such as 7% to 8%, such as 8% to 9%, such as from 9% to 10% by weight of fibrinogen.

[0131] In one embodiment, before mixing with an aqueous medium, the hemostatic composition comprises from 2% to 15% by weight, such as from 2.5% to 15% by weight, such as from 3% to 15% by weight, such as from about 3% by weight, such as from about 3.5% by weight, such as from about 4% by weight, such as from about 4.5% by weight, such as from about 5% by weight, such as from about 5.5% by weight, such as from about 6% by weight, such as from about 6.5% by weight, such as from about 7% by weight, such as from about 7.5% by weight, such as from about 8% by weight, such as from about 8.5% by weight, such as from about 9% by weight, such as from about 9.5% by weight, such as from about 10% by weight of fibrinogen.

[0132] Thrombin

[0133] Thrombin is a "trypsin-like" serine protease protein that is encoded by the F2 gene in humans. Prothrombin (coagulation factor II) is proteolytically cleaved in the coagulation cascade to form thrombin, which ultimately stops blood loss. Thrombin in turn acts as a serine protease that converts soluble fibrinogen into insoluble fibrin strands and catalyzes many other coagulation-related reactions. In the coagulation pathway, thrombin is used to convert factor XI to factor XIa, factor VIII to factor VIIIa, factor V to factor Va, and fibrinogen to fibrin.

[0134] In one embodiment, the thrombin is human thrombin.

[0135] In one embodiment, the thrombin is recombinant human thrombin.

[0136] In other embodiments, the origin of the thrombin is from a mammal other than a human, such as bovine thrombin.

[0137] In one embodiment, the thrombin is in the form of prothrombin.

[0138] In one embodiment, the thrombin is a dried thrombin composition. For example, the thrombin can be in the form of granules or powder. The dried thrombin composition can be prepared by any method known to those skilled in the art and is typically provided in a sterile form. Thus, in one embodiment, the dried thrombin composition is sterile.

[0139] Thrombin can also be coated on a biocompatible polymer in the form of granules. The coating of thrombin on the biocompatible polymer can be obtained by any method known in the art, such as by spray techniques, which can be carried out in any spray device. For example, a well-known method for spray-coating granules is the fluidized bed process. Thus, in one embodiment, thrombin is coated on biocompatible polymer granules to obtain a thrombin layer on the biocompatible polymer granules. In one embodiment, thrombin is sprayed onto the biocompatible polymer granules.

[0140] In some embodiments, the biocompatible polymer in the form of granules is coated with both thrombin and fibrinogen using known coating methods. Those skilled in the art will know how to perform such coatings in a spray device, such as in a fluidized bed process. For example, a first coating of fibrinogen or thrombin can be applied to the biocompatible polymer granules to obtain a first layer on the biocompatible polymer granules. Thereafter, a second coating of thrombin or fibrinogen can be applied to obtain a second layer on the first layer.

[0141] It may also combine thrombin and fibrinogen in the same layer. For example, a layer incorporating fibrinogen and thrombin is obtained by coating both thrombin and fibrinogen onto biocompatible polymer particles simultaneously. For example, thrombin and fibrinogen can be sprayed simultaneously from separate spray sources or from the same spray source.

[0142] In some embodiments, the medium for spraying thrombin and / or fibrinogen is a non-aqueous medium to prevent the catalytic reaction of thrombin on fibrinogen.

[0143] In one embodiment, a mixture of particles separately coated with thrombin and fibrinogen is obtained by: coating one subset of polymer particles with thrombin and another subset of polymer particles with fibrinogen, and then combining the two coated subsets at an appropriate ratio to obtain an optimal polymer:fibrinogen:thrombin ratio.

[0144] In one embodiment, the dried thrombin composition is prepared by spray drying or freeze drying.

[0145] In one embodiment, the dried thrombin composition is prepared by freeze drying.

[0146] In one embodiment, the dried thrombin composition contains less than 2% water, such as less than 1% water.

[0147] In one embodiment, the hemostatic composition contains thrombin in an amount of 400 to 4000 IU of thrombin per gram of biocompatible polymer, such as 400 to 600 IU per gram of biocompatible polymer, such as 600 to 800 IU, such as 800 to 1000 IU, such as 1000 to 1200 IU, such as 1200 to 1400 IU, such as 1400 to 1600 IU, such as 1600 to 1800 IU, such as 1800 to 2000 IU, such as 2000 to 2200 IU, such as 2200 to 2400 IU, such as 2400 to 2600 IU, such as 2600 to 2800 IU, such as 2800 to 3000 IU, such as 3000 to 3200 IU, such as 3200 to 3400 IU, such as 3400 to 3600 IU, such as 3600 to 3800 IU, such as 3800 to 4000 IU, such as 4000 to 4200 IU of thrombin.

[0148] In one embodiment, the hemostatic composition comprises thrombin in an amount of about 500 IU to 2500 IU of thrombin per gram of biocompatible polymer, such as about 600 IU to 2500 IU per gram of biocompatible polymer, such as about 700 IU to 2500 IU, such as about 800 IU to 2500 IU, such as about 900 IU to 2500 IU, such as about 1000 IU to 2500 IU, such as about 1100 IU to 2500 IU, such as about 1200 IU to 2500 IU, such as about 1300 IU to 2500 IU, such as about 1400 IU to 2500 IU, such as about 1500 IU to 2500 IU, such as about 1600 IU to 2500 IU, such as about 1700 IU to 2500 IU, such as about 1800 IU to 2500 IU, such as about 1900 IU to 2500 IU of thrombin, such as about 500 IU, such as about 1000 IU, such as about 1500 IU, such as about 2000 IU, such as about 2500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the composition comprises about 2000 IU of thrombin per gram of biocompatible polymer.

[0149] In one embodiment, the hemostatic composition comprises a ratio of thrombin to fibrinogen of 0.5 IU / mg to 5000 IU / mg, such as 1 IU / mg to 2000 IU / mg, such as 2 IU / mg to 1000 IU / mg, such as 2 IU / mg to 300 IU / mg, such as 2 IU / mg to 250 IU / mg, such as 2 IU / mg to 200 IU / mg, such as 2 IU / mg to 150 IU / mg.

[0150] In one embodiment, the hemostatic composition comprises 30 to 40 mg of fibrinogen and 500 to 1000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 30 to 40 mg of fibrinogen and 1000 to 1500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 30 to 40 mg of fibrinogen and 1500 to 2000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 30 to 40 mg of fibrinogen and 2000 to 2500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 40 to 50 mg of fibrinogen and 500 to 1000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 40 to 50 mg of fibrinogen and 1000 to 1500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 40 to 50 mg of fibrinogen and 1500 to 2000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 40 to 50 mg of fibrinogen and 2000 to 2500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 50 to 60 mg of fibrinogen and 500 to 1000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 50 to 60 mg of fibrinogen and 1000 to 1500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 50 to 60 mg of fibrinogen and 1500 to 2000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 50 to 60 mg of fibrinogen and 2000 to 2500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 60 to 70 mg of fibrinogen and 500 to 1000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 60 to 70 mg of fibrinogen and 1000 to 1500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 60 to 70 mg of fibrinogen and 1500 to 2000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 60 to 70 mg of fibrinogen and 2000 to 2500 IU of thrombin per gram of biocompatible polymer.

[0151] In one embodiment, the hemostatic composition comprises 35 mg of fibrinogen and 500 to 1000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 35 mg of fibrinogen and 1000 to 1500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 35 mg of fibrinogen and 1500 to 2000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 35 mg of fibrinogen and 2000 to 2500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 50 mg of fibrinogen and 500 to 1000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 50 mg of fibrinogen and 1000 to 1500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 50 mg of fibrinogen and 1500 to 2000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 50 mg of fibrinogen and 2000 to 2500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 70 mg of fibrinogen and 500 to 1000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 70 mg of fibrinogen and 1000 to 1500 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 70 mg of fibrinogen and 1500 to 2000 IU of thrombin per gram of biocompatible polymer. In one embodiment, the hemostatic composition comprises 70 mg of fibrinogen and 2000 to 2500 IU of thrombin per gram of biocompatible polymer.

[0152] In a particular embodiment, the hemostatic composition comprises:

[0153] a) about 1 g of a biocompatible polymer in particulate form, such as gelatin,

[0154] b) about 35 mg of fibrinogen,

[0155] c) about 2000 IU of thrombin.

[0156] In one embodiment, the hemostatic composition contains less than 10% water by weight, preferably less than 5% water by weight, preferably less than 1% water by weight.

[0157] Hydrophilic compound

[0158] In one embodiment, the hemostatic composition further comprises one or more hydrophilic compounds. Hydrophilic compounds typically contain polar or charged functional groups that render them soluble in water. Inclusion of one or more hydrophilic compounds in the hemostatic compositions of the present disclosure is believed to have a beneficial effect on thrombin stability and can improve the reconstitution efficiency of dry thrombin compositions. Hydrophilic compounds can also improve the consistency of the hemostatic composition.

[0159] In one embodiment, the hydrophilic compound is a hydrophilic polymer. The hydrophilic polymer can be natural or synthetic, linear or branched, and of any suitable length.

[0160] In one embodiment, the hydrophilic polymer is selected from the group consisting of polyethyleneimine (PEI), poly(ethylene glycol) (PEG), poly(ethylene oxide), poly(vinyl alcohol) (PVA), poly(styrene sulfonate) (PSS), poly(acrylic acid) (PAA), poly(allylamine hydrochloride), and poly(vinyl acid).

[0161] In one embodiment, the hydrophilic polymer is poly(ethylene glycol) (PEG).

[0162] In one embodiment, the hydrophilic compound is selected from the group consisting of cetylpyridinium chloride, docusate sodium, glycine, hypromellose, phthalates, lecithin, phospholipids, poloxamer, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, polyvinyl alcohol, sodium lauryl sulfate, sorbitan esters (sorbitan fatty acid esters), and glyceryl trioctanoate.

[0163] In a preferred embodiment, the hydrophilic compound is a polyol. Thus, according to one embodiment of the invention, one or more polyols can be included in the hemostatic composition. Polyols can enhance the reconstitution rate of dry thrombin compositions, stabilize thrombin activity, and play a role in ensuring the optimal consistency of the hemostatic composition.

[0164] As defined herein, a polyol is a compound having multiple hydroxyl functional groups. Polyols include sugars (monosaccharides, disaccharides, and polysaccharides), sugar alcohols, and their derivatives. Sugar alcohols are particularly preferred.

[0165] Monosaccharides include, but are not limited to, glucose, fructose, galactose, xylose, and ribose.

[0166] Disaccharides include, but are not limited to, sucrose (sucrose / saccharose), lactulose, lactose, maltose, trehalose, and cellobiose.

[0167] Polysaccharides include, but are not limited to, starch, glycogen, cellulose, and chitin.

[0168] Sugar alcohols, also known as polyols, are the hydrogenated forms of carbohydrates in which the carbonyl group (aldehyde or ketone, reducing sugar) has been reduced to a primary or secondary hydroxyl group (hence the alcohol). Sugar alcohols have the general formula H(HCHO)n+1H, while sugars have H(HCHO)nHCO. Some common sugar alcohols useful in the methods of the present disclosure include, but are not limited to: ethylene glycol (2 carbons), glycerol (3 carbons), erythritol (4 carbons), threitol (4 carbons), arabitol (5 carbons), xylitol (5 carbons), ribitol (5 carbons), mannitol (6 carbons), sorbitol (6 carbons), dulcitol (6 carbons), fucitol (6 carbons), iditol (6 carbons), inositol (6 carbons; cyclic sugar alcohol), volemitol (7 carbons), isomalt (12 carbons), maltitol (12 carbons), lactitol (12 carbons), polyglycitol.

[0169] In one embodiment, the hemostatic composition comprises a single hydrophilic compound, such as a single polyol.

[0170] In one embodiment of the present invention, the hemostatic composition comprises more than one hydrophilic compound, such as two, three, four, five, six or even more different hydrophilic compounds.

[0171] In a preferred embodiment, the hydrophilic compound is a polyol.

[0172] In one embodiment of the present invention, the hemostatic composition comprises two polyols, such as mannitol and glycerol or trehalose and ethylene glycol.

[0173] In one embodiment of the present invention, the hemostatic composition comprises one or more sugar alcohols, such as one or more sugar alcohols selected from the group consisting of: ethylene glycol, glycerol, erythritol, threitol, arabitol, xylitol, ribitol, mannitol, sorbitol, dulcitol, fucitol, iditol, inositol, volemitol, isomalt, maltitol, lactitol, polyglycitol.

[0174] In one embodiment, the hemostatic composition comprises one or more sugar alcohols and one or more sugars, such as one sugar alcohol and one sugar.

[0175] In one embodiment, the hemostatic composition comprises one sugar alcohol and optionally one or more additional hydrophilic compounds, such as one or more polyols, which may be sugar alcohols or sugars.

[0176] In one embodiment, the hemostatic composition does not contain sugar as the sole polyol.

[0177] In one embodiment of the present invention, the hemostatic composition comprises mannitol.

[0178] In one embodiment of the present invention, the hemostatic composition comprises sorbitol.

[0179] In one embodiment of the present invention, the hemostatic composition comprises glycerol.

[0180] In one embodiment of the present invention, the hemostatic composition comprises trehalose.

[0181] In one embodiment of the present invention, the hemostatic composition comprises a diol, such as propylene glycol.

[0182] In one embodiment of the present invention, the hemostatic composition comprises xylitol.

[0183] In one embodiment of the present invention, the hemostatic composition comprises maltitol.

[0184] In one embodiment of the present invention, the hemostatic composition comprises sorbitol.

[0185] In one embodiment, the hemostatic composition comprises from 0.01 g to 0.5 g of a hydrophilic compound per gram of biocompatible polymer, such as from 0.01 g to 0.4 g per gram of biocompatible polymer, such as from 0.01 to 0.3 g, such as from 0.01 to 0.2 g, such as from 0.01 to 0.1 g, such as from 0.01 to 0.05 g of a hydrophilic compound.

[0186] Other bioactive agents

[0187] In one embodiment of the present invention, the hemostatic composition comprises one or more other bioactive agents capable of stimulating hemostasis, wound healing, bone healing, tissue healing, and / or tendon healing.

[0188] In one embodiment, the hemostatic composition comprises one or more other bioactive agents that stimulate the healing of bone and / or tendon and / or tissue, such as one or more growth factors selected from the group consisting of matrix metalloproteinase (MMP), insulin-like growth factor 1 (IGF-I), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and transforming growth factor β (TGF-β).

[0189] In one embodiment, the hemostatic composition comprises one or more bone morphogenetic proteins (BMPs). Bone morphogenetic proteins (BMPs) are a subgroup of the TGF-β superfamily. Bone morphogenetic proteins (BMPs) are a group of growth factors, also known as cytokines and metabologens. BMPs were originally discovered for their ability to induce bone and cartilage formation and are now considered to constitute a key set of morphogenetic signals that coordinate the tissue architecture of the entire body.

[0190] In one embodiment, the hemostatic composition comprises one or more matrix metalloproteinases (MMPs). MMPs are zinc-dependent endopeptidases. MMPs play a very important role in the degradation and remodeling of the extracellular matrix (ECM) during the wound healing process. Certain MMPs, including MMP-1, MMP-2, MMP-8, MMP-13, and MMP-14, have collagenase activity, meaning that, unlike many other enzymes, they are able to degrade type I collagen fibrils.

[0191] These growth factors all have different roles during the healing process. IGF-1 increases collagen and proteoglycan production during the first stage of inflammation, and PDGF is also present during the early stages after injury and promotes the synthesis of other growth factors as well as DNA synthesis and cell proliferation. Three isoforms of TGF-β (TGF-β1, TGF-β2, TGF-β3) are known to play a role in wound healing and scar formation. VEGF is well known to promote angiogenesis and induce endothelial cell proliferation and migration.

[0192] In one embodiment, the hemostatic composition of the present disclosure comprises sheets or particles of the extracellular matrix (ECM). The ECM is the extracellular part of animal tissue, which generally provides structural support for animal cells and performs various other important functions. The ECM has been shown to have highly beneficial effects in healing because it promotes functional tissue regeneration.

[0193] The variety of other bioactive agents that can be used in combination with the hemostatic composition of the present invention is very broad. Generally, bioactive agents that can be administered via the hemostatic composition of the present invention include, but are not limited to, anti-infective agents, such as antibiotics and antiviral agents; analgesics and analgesic combinations; antihelminthics; anti-arthritics; anticonvulsants; antidepressants; antihistamines; anti-inflammatory agents; anti-migraine preparations; anti-tumor drugs; anti-Parkinson's drugs; antipsychotics; antipyretics; antispasmodics; anticholinergics; sympathomimetics; xanthine derivatives; cardiovascular preparations, including calcium channel blockers and β-blockers, such as pindolol and antiarrhythmic drugs; antihypertensive agents; diuretics; vasodilators, including vasodilators of general coronary, peripheral and cerebral blood vessels; central nervous system stimulants; hormones, such as estradiol and other steroids, including corticosteroids; immunosuppressants; muscle relaxants; parasympathetic blockers; psychostimulants; proteins, polysaccharides, glycoproteins or lipoproteins of natural origin or genetically engineered; oligonucleotides, antibodies, antigens, cholinergic agents, chemotherapeutic agents, radioactive agents, bone inductors, heparin neutralizers for cell growth inhibition, procoagulants and hemostatic agents, such as fibrin, fibronectin, heparinase, factor X / Xa, factor VII / VIla, factor VIII / VIIIa, factor IX / IXa, factor XI / XIa, factor XII / XIIa, factor XIII / XIIIa, tissue factor, batroxobin, ancrod, ecarin, von Willebrand Factor, platelet surface glycoproteins, vasopressin, vasopressin analogs, epinephrine, selectins, procoagulant venoms, plasminogen activator inhibitors, platelet activators and synthetic peptides having hemostatic activity.

[0194] Other compounds

[0195] The hemostatic composition of the present invention may further comprise one or more of the following: DMSO (dimethyl sulfoxide) and / or 2-methyl-2,4-pentanediol (MPD).

[0196] In one embodiment, the hemostatic composition of the present disclosure comprises one or more antimicrobial agents, such as one or more antibacterial agents.

[0197] In one embodiment, the hemostatic composition of the present disclosure comprises benzalkonium chloride (BAC).

[0198] In one embodiment, the hemostatic composition of the present disclosure does not comprise an antimicrobial agent.

[0199] In one embodiment, the hemostatic composition further comprises an extrusion enhancer, i.e., a compound that facilitates extrusion of the paste from a syringe.

[0200] It has previously been demonstrated that providing an appropriate amount of certain extrusion enhancers, such as albumin, enables the use of higher gelatin concentrations because it reduces the amount of force required to extrude the gelatin paste composition from, for example, a syringe. Using a higher gelatin concentration in turn can improve the hemostatic properties of such products. It is necessary to provide an appropriate amount of the extrusion enhancer. The amount is preferably high enough to obtain an extrusion effect, i.e., a flowable paste can be obtained even for a relatively high amount of biocompatible polymer (e.g., cross-linked gelatin), such that the hemostatic composition can be accurately applied by a surgeon using, for example, a syringe containing an applicator tip; on the other hand, the amount should be low enough to prevent potential negative functional properties of the hemostatic composition.

[0201] The extrusion enhancer is preferably albumin, particularly human serum albumin.

[0202] In one embodiment, the hemostatic composition in paste form (i.e., after reconstitution with an aqueous medium) contains an extrusion enhancer, such as albumin, in an amount of from about 0.1% to about 10%, such as from about 0.2% to about 8%, for example from about 0.3% to about 7%, preferably from about 0.5% to about 5%, such as from about 1% to about 4%.

[0203] In one embodiment, the hemostatic composition of the present disclosure contains only trace amounts of albumin, such as less than 0.1%, for example less than 0.01%, such as less than 0.001%, for example less than 0.0001%.

[0204] Method for preparing a hemostatic composition

[0205] In one aspect, the present disclosure provides a method for preparing a hemostatic composition, which comprises the following steps:

[0206] a) providing a hemostatic composition as described herein, and

[0207] b) adding a certain amount of aqueous medium to the hemostatic composition of a).

[0208] In one embodiment, the amount of aqueous medium added is 2 to 12 mL of aqueous medium per gram of the hemostatic composition provided in step a), such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 mL of aqueous medium.

[0209] In one embodiment, the amount of aqueous medium added is 4 to 7 mL, such as 5 to 6 mL, per gram of the hemostatic composition provided in step a).

[0210] In one aspect, the present disclosure provides a method for preparing a hemostatic composition, which comprises the following steps:

[0211] a) Providing a hemostatic composition comprising a biocompatible polymer in particulate form and fibrinogen, wherein the biocompatible polymer and fibrinogen are as described herein, and

[0212] b) Adding an amount of an aqueous medium comprising thrombin.

[0213] In one embodiment, the present disclosure provides a method of preparing a hemostatic composition, comprising the steps of:

[0214] a) Providing a hemostatic composition comprising a biocompatible polymer in particulate form, fibrinogen, and an aqueous medium, wherein the biocompatible polymer, fibrinogen, and aqueous medium are as described herein, and

[0215] b) Adding an amount of an aqueous medium comprising thrombin.

[0216] The components are mixed with the aqueous medium by conventional means, such as by transferring between two connected syringes, to form a paste.

[0217] The hemostatic composition obtained by the method described herein is suitable for hemostasis and / or wound healing.

[0218] In one aspect, the present disclosure provides a hemostatic composition obtained by the method described herein. The hemostatic composition obtained by the method described herein is preferably a flowable paste composition.

[0219] Surprisingly, the inventors have demonstrated that a hemostatic paste composition comprising a biocompatible polymer, thrombin, and fibrinogen remains flowable for more than 1.5 hours after reconstitution, i.e., after mixing with an aqueous medium, as shown in the examples. This finding was highly unexpected because current sealants comprising thrombin and fibrinogen readily react and form fibrin clots that would clog the device upon combination and thus cannot be deployed from a syringe for an extended period of time after the components are combined.

[0220] In one embodiment, the hemostatic composition remains flowable for at least 4 hours after adding the aqueous medium, such as at least 2 hours, such as at least 90 minutes, such as at least 60 minutes, such as at least 30 minutes.

[0221] In one embodiment, the hemostatic composition can be deployed from a syringe for at least 4 hours after adding the aqueous medium, such as at least 2 hours, such as at least 90 minutes, such as at least 60 minutes, such as at least 30 minutes.

[0222] In one embodiment, the hemostatic composition is a paste. Thus, in one embodiment, the amount of aqueous medium added to a biocompatible polymer in particulate form, such as cross-linked gelatin particles, is an amount suitable for forming a paste.

[0223] In one embodiment, the hemostatic composition has a consistency in the range of from about 100 g·sec to about 15,000 g·sec, such as from about 500 g·sec to about 8000 g·sec, for example from about 1000 g·sec to about 5000 g·sec, such as from about 1500 g·sec to about 3000 g·sec.

[0224] In one embodiment, the hemostatic composition has a consistency of less than about 5000 g·sec, for example less than about 4000 g·sec, such as less than about 3000 g·sec, for example less than about 2000 g·sec.

[0225] The consistency can be measured using a texture analyzer (TA.XT.plus, Stable micro systems) under the following TA settings:

[0226] Test mode Compression Pre-test speed 5.00 mm / sec Test speed 0.5 mm / sec Post-test speed 10 mm / sec Distance 30.0 mm Trigger type Automatic Trigger force 4.0g Probe P / 0.5R 1 / 2” diameter cylinder

[0227] For a medical paste to be discharged from a syringe and an applicator tube, it should be flowable when subjected to a force suitable for a syringe. Thus, the term "flowable paste" means a paste having a viscosity that promotes stable flow when subjected to a force suitable for a syringe. The flowability of the paste can be measured, for example, at 25 °C - 30 °C and 65% - 75% relative humidity.

[0228] In one embodiment of the present disclosure, the hemostatic composition in paste form has a viscosity in the range of from about 500 Pa·s to about 8000 Pa·s, such as from about 500 to about 7000 Pa·s, such as from about 500 Pa·s to about 6000 Pa·s, such as from about 600 Pa·s to about 5000 Pa·s, such as from about 700 Pa·s to about 4000 Pa·s, such as from 800 Pa·s to about 3000 Pa·s, such as from about 1000 Pa·s to about 2500 Pa·s, such as 1500 Pa·s. In one embodiment, the hemostatic composition has a viscosity of less than about 2500 Pa·s.

[0229] The viscosity of the paste can be measured by a rheometer, and preferably by a rotational shear-based rheometer. The viscosity can be measured using a Discovery Hybrid Rheometer (DHR-1) from Waters TA Instruments under controlled stress and the following measurement conditions: time sweep oscillatory measurement mode, oscillatory strain of 1%, angular frequency of 1 rad / s, 20 mm plate as the upper geometry diameter, and gap size of 1.25 mm. The measurement can be carried out at a temperature between 25°C and 30°C or thereabouts, and preferably at a temperature of 25°C and a relative humidity of 65% - 75%.

[0230] Thus, in one embodiment, the present disclosure provides a hemostatic composition comprising:

[0231] a) one or more biocompatible polymers in particulate form,

[0232] b) fibrinogen,

[0233] c) thrombin, and

[0234] d) an aqueous medium.

[0235] Preferably, the hemostatic composition comprises an amount of aqueous medium sufficient to form a flowable paste.

[0236] In one embodiment, the aqueous medium is selected from the group consisting of: water, saline, calcium chloride solution, and buffered aqueous media. The water can be WFI (Water for Injection). In one embodiment, the aqueous medium is selected such that the reconstituted paste product is substantially isotonic. The aqueous medium is preferably sterile.

[0237] In one embodiment, the aqueous medium contains calcium ions.

[0238] In one embodiment, the aqueous medium of the present disclosure is a saline solution.

[0239] In one embodiment, the aqueous medium is a calcium chloride solution.

[0240] In other embodiments, the aqueous medium is water.

[0241] The aqueous medium can also be a buffered aqueous medium. Any suitable buffer known to those skilled in the art can be used, such as one or more buffers selected from the group consisting of: sodium citrate; citric acid; acetic acid, sodium acetate; KH 2 HPO 4 、KH 2 PO 4 ;Na 2 HPO 4 、NaH 2 PO 4; CHES; borax, sodium hydroxide; TAPS; bicine; Tris; tricine; TAPSO; HEPES; TES; MOPS; PIPES; cacodylate; SSC; MES or other buffers. The pH of the buffered aqueous medium should be suitable for producing a hemostatic composition intended for human use and can be determined by those skilled in the art.

[0242] The amount of the aqueous medium is an amount sufficient to provide the composition in the form of a paste, such as 2 to 12 mL per gram of biocompatible polymer, such as 4 to 10 mL per gram of biocompatible polymer, such as 4 to 8 mL, such as 4 to 7 mL, such as 5 to 6 mL of aqueous medium.

[0243] In one embodiment, the present disclosure provides a hemostatic composition comprising:

[0244] a) one or more biocompatible polymers in particulate form, in an amount of 7% to 34% by weight,

[0245] b) fibrinogen, in an amount of 0.008% to 5% by weight,

[0246] c) thrombin, in an amount of 7 to 1700 IU per gram, and

[0247] d) an aqueous medium.

[0248] In one embodiment, the hemostatic composition comprises a biocompatible polymer in particulate form in an amount of about 7% to 20% by weight, such as about 9% to 19%, for example about 11% to 18%, such as about 12% - 17%, for example about 14% - 17%.

[0249] In one embodiment, the hemostatic composition comprises a biocompatible polymer in particulate form in an amount of about 7% to 20% by weight, such as about 10% to 20%, for example about 11% to 20%, such as about 12% to 20%, for example about 13% to 20%, such as about 14% to 20%, for example about 14% to 19%, such as about 14% to 18%, for example about 14% to 17%.

[0250] In one embodiment, the hemostatic composition comprises a biocompatible polymer in particulate form in an amount of about 13% to 15%, such as about 14%. In another embodiment, the hemostatic composition comprises a biocompatible polymer in particulate form in an amount of about 16% to 18% by weight, such as about 17%.

[0251] In one embodiment, the hemostatic composition comprises from about 60% to about 93% water, such as from about 70% to about 90% water, such as from about 75% to about 90% water, such as from about 80% to about 90% water.

[0252] In one embodiment, the hemostatic composition in the form of a flowable paste comprises from 0.01% to 2.5% by weight, such as from 0.1% to 2% by weight, such as from 0.2% to 2% by weight, such as from 0.3% to 2% by weight, such as from 0.4% to 2% by weight, such as from 0.4% to 1.8% by weight, such as from 0.4% to 1.7% by weight, such as from 0.4% to 1.6% by weight, such as from 0.4% to 1.5% by weight of fibrinogen. In one embodiment, the hemostatic composition comprises from 0.3% to 1.2% by weight, such as about 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0% or about 1.1% by weight of fibrinogen. In one embodiment, the hemostatic composition comprises from 0.3% to 1.2% by weight of fibrinogen.

[0253] In one embodiment, the hemostatic composition in the form of a flowable paste comprises from 16 to 900 IU / g, such as from 100 to 700 IU / g, such as from 100 to 200 IU / g, such as from 100 to 200 IU / g, such as from 200 to 300 IU / g, such as from 300 to 400 IU / g, such as from 400 to 500 IU / g, such as from 500 to 600 IU / g or from 600 to 700 IU / g of thrombin. In one embodiment, the hemostatic composition comprises from 280 to 350 IU / g of the hemostatic composition of thrombin.

[0254] In one embodiment, the hemostatic composition as described herein is sterile.

[0255] Container

[0256] In one embodiment, the hemostatic composition as described herein is provided in a first container and the aqueous solution is provided in a second container, and the containers can form a final hemostatic paste composition to be applied to the bleeding site after connection.

[0257] The first and second containers can be made of any suitable material, such as plastic, glass, ceramic, plastic or metal, such as stainless steel. Examples of suitable plastic materials include, but are not limited to, polyethylene, polypropylene, polystyrene, polyvinyl chloride and polytetrafluoroethylene (PTFE).

[0258] In one embodiment, the hemostatic composition is provided in a container selected from a syringe, vial, jar, tube, tray or cartridge.

[0259] In a preferred embodiment, the first container containing the hemostatic composition is a medical delivery device suitable for dispensing a flowable hemostatic composition to a patient in need, such as a syringe, such as a single-use plastic syringe.

[0260] The first container is generally made of a material suitable for chemical surface sterilization without affecting the contents of the container. For example, the first container can be made of a material impermeable to ethylene oxide, such as made of metal, glass, or ethylene oxide-impermeable plastic.

[0261] In one embodiment, an aqueous solution is provided in a second container, which can be selected from a syringe, vial, bottle, tube, tray, or cartridge.

[0262] In a preferred embodiment, the container containing the aqueous solution is a medical delivery device suitable for dispensing a flowable hemostatic composition to a patient in need, such as a syringe. In one embodiment, the second container is a single-use plastic syringe.

[0263] In one embodiment, the first and second containers are interconnected. The connector portion can be a standard type of connector portion, such as a Luer lock or Luer slip connector. The connector portion can have a threaded portion for secure connection with a mating connector. The size of the Luer lock or Luer slip connector can be capable of changing the ability to mix air into the hemostatic composition during the mixing of the hemostatic composition and the aqueous medium. In addition, the size of the Luer lock or Luer slip connector can be capable of affecting the consistency of the hemostatic composition.

[0264] In one embodiment, the connector portion includes a static mixer. The size of the static mixer can be capable of changing the ability to mix air into the hemostatic composition during the mixing of the hemostatic composition and the aqueous medium. In addition, the size of the static mixer can be capable of affecting the consistency of the hemostatic composition.

[0265] Sterilization

[0266] The hemostatic composition according to the present disclosure is preferably sterile.

[0267] Thus, in one embodiment, the hemostatic composition described herein is a sterile hemostatic composition.

[0268] Any suitable sterilization technique known in the art can be used. Sterilization refers to any process that effectively kills or eliminates infectious agents (such as fungi, bacteria, viruses, prions, and spores, etc.). Sterilization can be achieved, for example, by applying heat, chemicals, and / or irradiation.

[0269] Sterilization can be achieved by heat sterilization, including autoclaving (using high-temperature steam) and dry heat.

[0270] Sterilization can also be achieved by irradiation (such as ionizing irradiation) to provide sterility to the composition. Such irradiation can include electron beam (β irradiation), X-rays, γ and β rays, UV light, and subatomic particles. The irradiation levels and conditions (including time) for sterilization are those that provide a sterile composition. The sterilization conditions are similar to those currently used in the art and can be determined by a person skilled in the art.

[0271] Sterilization can be carried out by chemical sterilization, such as by using ethylene oxide gas, ozone, nitrogen dioxide, chlorine bleach, glutaraldehyde, formaldehyde, ortho-phthalaldehyde, hydrogen peroxide, and / or peracetic acid.

[0272] Sterile methods can also be used to prepare the hemostatic composition or the biocompatible polymer in particulate form.

[0273] Medical uses

[0274] The present disclosure further relates to a hemostatic composition as described herein or obtained by the methods of the present disclosure, which is used to promote hemostasis and / or wound, bone, tendon, and / or tissue healing in an individual in need thereof.

[0275] In one embodiment, the present disclosure provides a hemostatic composition as described herein, which is used to promote hemostasis in an individual in need thereof. In one embodiment, the present disclosure provides a hemostatic composition as described herein, which is used to promote wound healing in an individual in need thereof.

[0276] The hemostatic composition of the present disclosure can be used, for example, in a series of surgical procedures that require control of bleeding. The hemostatic composition is preferably applied to the patient in the form of a paste, and the paste conforms to the irregular surface and can thus be used to provide rapid hemostasis on rough or uneven surfaces where hemostatic sponges are not effective.

[0277] Due to its superior hemostatic and adhesive properties, the hemostatic composition as disclosed herein is particularly suitable for minimally invasive / robotic surgery, where manual compression is impractical or impossible and / or where classical hemostatic fibrin sealants are ineffective. For example, the hemostatic composition in the form of a paste as disclosed herein can be sprayed onto the bleeding surface during minimally invasive surgery to provide a patch-like hemostatic composition that adheres well to the bleeding site and provides effective hemostasis without the need for compression.

[0278] The hemostatic composition as disclosed herein has also been shown to be capable of controlling severe bleeding. Thus, in one embodiment, the hemostatic composition as disclosed herein can be used to treat severe bleeding classified as grade 4 or 5 bleeding.

[0279] Generally, hemostatic pastes are prepared directly by a practicing physician at the surgical site as needed by adding a liquid to a container (such as a syringe) containing a certain amount of hemostatic product. The hemostatic product can be pre-wetted with the liquid or be substantially dry (e.g., a free-flowing powder). Thus, the paste is typically prepared under conditions of extreme pressure, and thus it is crucial that the process of preparing the paste is simple and fast to ensure that bleeding is stopped as quickly as possible and without error when preparing the paste, such that the nurse can focus on the needs of the surgeon rather than on preparing the hemostatic agent. It is also important that the consistency of the paste is suitable for use as a hemostatic paste and that the consistency of the product is independent of the preparation batch and over time. Currently available flowable paste products ( and ) require separate reconstitution of thrombin and then mechanical mixing of the reconstituted thrombin solution with a biocompatible polymer by passing the biocompatible polymer and the liquid back and forth between two connected syringes multiple times to obtain a substantially homogeneous paste. The reconstitution of thrombin is time-consuming and error-prone, which are two undesirable factors in the operating room (OR) environment. These products are typically pre-prepared in the operating room before surgery for use during surgery, and unused products are typically discarded, resulting in unnecessarily high operating room costs.

[0280] Since the hemostatic composition in a dry, storage-stable form of the present disclosure already includes thrombin and fibrinogen, a hemostatic paste for application to a patient can be more easily prepared because there is no need for separate reconstitution and addition of, for example, thrombin. To obtain the hemostatic composition of the present disclosure in paste form, a suitable amount of aqueous medium can simply be added to a container containing the hemostatic composition (in dry form) and the contents can be mixed by transferring them back and forth between two interconnected syringes multiple times, thereby forming a ready-to-use hemostatic paste.

[0281] A significant advantage of the compositions and methods of the present invention is that they allow for better control of the consistency of the hemostatic composition while providing superior hemostatic efficacy. This is very valuable in the operating room where effective and as-fast-as-possible hemostasis is required.

[0282] Another advantage of the compositions and methods of the present disclosure is that, compared to current hemostatic flowable kits, kits consisting of fewer components can be prepared. Preparation of the flowable paste composition in the operating room requires only a dry, storage-stable hemostatic composition contained within a first container (such as a syringe), and a second container (such as a syringe) containing an aqueous solution. After connecting and mixing the two, a ready-to-use flowable paste containing all the agents required for efficient hemostasis is formed. Thus, no additional syringes, vial adapters, needles, and mixing bowls are required. This means that manufacturing costs can be reduced and good patient safety is also ensured, since there are fewer components for operating room staff to track during surgery. The needleless preparation of the hemostatic agent also ensures the safety of operating room staff.

[0283] In one embodiment, the present disclosure relates to a method of stopping bleeding / promoting hemostasis in an individual in need by applying a hemostatic composition as disclosed herein to a bleeding site. When applied, the hemostatic composition is preferably in the form of a flowable paste.

[0284] The hemostatic compositions of the present disclosure can be used in any type of surgery, including general surgery, cardiothoracic surgery, vascular surgery, plastic surgery, pediatric surgery, colorectal surgery, transplant surgery, surgical oncology, trauma surgery, endocrine surgery, breast surgery, dermatologic surgery, otolaryngology, gynecology, oral and maxillofacial surgery, dental surgery, plastic reconstructive surgery, neurosurgery, ophthalmology, podiatric surgery, urology. The hemostatic compositions are particularly suitable for minimally invasive / robotic surgery, where manual pressure is impractical or impossible.

[0285] In one embodiment, the present disclosure relates to a method of promoting wound healing in an individual in need by applying a hemostatic composition as disclosed herein to a wound.

[0286] "Wound" broadly refers to an injury to the skin and / or underlying (subcutaneous) tissue, which can originate in different ways (such as pressure ulcers from prolonged bed rest and wounds induced by trauma) and have different characteristics. Depending on the depth of the wound, the wound can be classified into one of four grades: i) Grade I: wounds limited to the epithelium; ii) Grade II: wounds extending into the dermis; iii) Grade III: wounds extending into the subcutaneous tissue; and iv) Grade IV (or full-thickness wound): wounds with bone exposure (such as bone pressure points, such as the greater trochanter or the sacrum). The present disclosure relates to treating any type of wound mentioned above using the hemostatic compositions of the present disclosure.

[0287] Treatment of a wound can in principle cause wound healing or accelerate wound healing. Accelerated healing can be, for example, the result of administering substances that promote wound healing. Optionally, wound healing can be promoted by preventing bacterial or viral infections or by reducing the risk of infections that would otherwise prolong the wound treatment process.

[0288] In one embodiment, the present disclosure relates to a method for promoting the healing of bone and / or tendon in an individual in need thereof by applying a hemostatic composition as disclosed herein to an injured bone / tendon.

[0289] As used herein, an "individual" can be any mammal, including but not limited to mammals of the order Rodentia (such as mice and hamsters) and mammals of the order Lagomorpha (such as rabbits). Preferably, the mammal is from the order Carnivora, including felines (cats) and canines (dogs). More preferably, the mammal is from the order Artiodactyla, including bovines (cows) and suids (pigs); or the order Perissodactyla, including equines (horses). Most preferably, the mammal is of the order Primates, suborder Anthropoidea or monkeys (monkeys) or suborder Hominoidea (humans and apes). Particularly preferred mammals are humans.

[0290] In one embodiment, the present disclosure relates to a hemostatic composition as disclosed herein for treating a wound, e.g., for stopping bleeding or for promoting wound healing.

[0291] Hemostatic kit

[0292] The present disclosure also relates to a hemostatic kit comprising a dry hemostatic composition as described herein and an aqueous solution or aqueous medium such that upon mixing, a hemostatic composition suitable for hemostasis will be formed.

[0293] Thus, in one embodiment, the present disclosure relates to a hemostatic kit comprising:

[0294] a) a first syringe containing a dry hemostatic composition as described herein,

[0295] b) a second syringe containing an aqueous medium; and

[0296] c) an optional outer package,

[0297] wherein the two syringes are interconnectable.

[0298] Thus, in one embodiment, the present disclosure relates to a hemostatic kit comprising:

[0299] a) a first syringe containing a dry hemostatic composition as described herein, the dry hemostatic composition comprising one or more biocompatible polymers in particulate form (such as crosslinked gelatin particles), thrombin, and fibrinogen,

[0300] b) a second syringe containing an aqueous medium; and

[0301] c) an optional outer package,

[0302] Two of the syringes are interconnectable.

[0303] In one embodiment, the kit further comprises one or more applicator tips.

[0304] The kit may optionally contain instructions for use of the kit.

[0305] The components of the hemostatic kit may be as described elsewhere herein.

[0306] In one embodiment, the kit comprises an outer package. The outer package is generally made of a flexible, semi-rigid or rigid material and generally consists of materials such as plastics, aluminum foils and / or plastic laminates, wherein the plastics are optionally selected from the group consisting of PET, PETG, PE, LLDPE, CPP, PA, PETP, METPET, Tyvek, and are optionally bonded or co-extruded with an adhesive (such as polyurethane).

[0307] In one embodiment, the outer package is an aluminum foil outer package.

[0308] The outer package preferably forms a complete moisture barrier.

[0309] The outer package preferably is capable of withstanding a sterilization process (such as sterilization by radiation).

[0310] Item

[0311] 1. A hemostatic composition comprising:

[0312] a) one or more biocompatible polymers in particulate form,

[0313] b) fibrinogen in an amount of 1 to 150 mg per gram of biocompatible polymer, and

[0314] c) thrombin in an amount of 100 to 5000 IU per gram of biocompatible polymer.

[0315] 2. The hemostatic composition according to item 1, wherein the one or more biocompatible polymers comprise or consist of powder particles that are substantially insoluble in an aqueous medium.

[0316] 3. The hemostatic composition according to any one of the preceding items, wherein the one or more biocompatible polymers in particulate form comprise or consist of crosslinked gelatin particles.

[0317] 4. The hemostatic composition according to any one of the preceding items, wherein the composition comprises 5 to 100 mg of fibrinogen per gram of biocompatible polymer, such as 20 to 80 mg per gram of biocompatible polymer, such as about 35 mg or such as about 70 mg of fibrinogen.

[0318] 5. The hemostatic composition according to any one of the preceding items, wherein the composition comprises 400 to 4000 IU of thrombin per gram of biocompatible polymer, such as 300 to 3000 IU per gram of biocompatible polymer, such as 500 to 2500 IU, such as about 2000 IU of thrombin.

[0319] 6. The hemostatic composition according to any one of the preceding items, wherein the composition comprises:

[0320] a) about 1 g of a biocompatible polymer in particulate form, such as gelatin,

[0321] b) about 20 to 100 mg of fibrinogen,

[0322] c) about 2000 IU of thrombin.

[0323] 7. The hemostatic composition according to any one of the preceding items, wherein the composition contains less than 10% by weight of water, preferably less than 5% by weight of water, preferably less than 1% by weight of water.

[0324] 8. A method for preparing a hemostatic composition, comprising the steps of:

[0325] a) providing a hemostatic composition according to any one of items 1 to 7, and

[0326] b) adding a quantity of aqueous medium to the hemostatic composition of a).

[0327] 9. A hemostatic composition obtained by the method according to item 8.

[0328] 10. The hemostatic composition according to item 9, wherein the composition remains flowable for at least 4 hours, such as at least 2 hours, such as at least 1 hour, such as at least 30 minutes after addition of the aqueous medium.

[0329] 11. A hemostatic composition, comprising:

[0330] a) one or more biocompatible polymers in particulate form, in an amount of 14% to 18% by weight of the hemostatic composition,

[0331] b) fibrinogen, in an amount of 0.3% to 1.2% by weight of the hemostatic composition,

[0332] c) thrombin, in an amount of 150 to 700 IU per gram of the hemostatic composition, and

[0333] d) an aqueous medium,

[0334] Wherein the hemostatic composition is a paste.

[0335] 12. The hemostatic composition according to any one of items 1 to 7 or 9 to 11, which is used to promote hemostasis and / or wound, bone, tendon and / or tissue healing in an individual in need thereof.

[0336] 13. A container comprising the hemostatic composition according to any one of items 1 to 7 or according to any one of items 9 to 11.

[0337] 14. The container according to item 13, wherein the container is an applicator, such as a syringe.

[0338] 15. A kit comprising:

[0339] a) A first container containing the composition according to any one of items 1 to 7,

[0340] b) A second container containing an aqueous medium; and

[0341] c) Optionally, an outer package,

[0342] wherein the two containers are interconnected. Examples

[0343] Example 1: Deployability of a paste composition containing thrombin, fibrinogen and a biocompatible polymer

[0344] Objective

[0345] To study the time for which the composition according to the present disclosure remains deployable from a syringe.

[0346] Materials

[0347] 5 mL of a gelatin paste containing 1 g of cross-linked gelatin particles and human recombinant fibrinogen in a 10 mL syringe (first syringe). Three different amounts of fibrinogen were tested: 7, 35 and 70 mg.

[0348] 1 mL of a 2000 IU human recombinant thrombin solution in a 10 mL syringe (second syringe).

[0349] The gelatin particles used in the paste were obtained from ground cross-linked gelatin sponges (Spongostan).

[0350] Method

[0351] The two syringes were interconnected via a Luer lock, and the paste was transferred back and forth between the first syringe and the second syringe until mixed.

[0352] The mixed fibrinogen-containing paste contains approximately 14% w / w gelatin, approximately 285 IU of thrombin per gram of paste, and approximately 0.1% w / w (7 mg), 0.5% w / w (35 mg), or 1% w / w (70 mg) of fibrinogen, respectively.

[0353] The syringe was left at room temperature for the period indicated in Table 1, and the clogging of the paste in the syringe and the ability to deploy the paste from the syringe via manual pressure on the plunger were evaluated.

[0354] Results

[0355] As can be seen in Table 1, the compositions with different amounts of fibrinogen according to the present disclosure remained deployable from the syringe for at least 90 minutes after mixing. In contrast, the composition without the biocompatible polymer clogged in the syringe and could not be deployed at any time point. A paste containing gelatin, thrombin, and 105 mg of fibrinogen per gram of gelatin was also tested (data not shown). The paste containing 105 mg of fibrinogen was deployable from the syringe, but the force required to deploy the sample was found to be significantly increased compared to the samples with less fibrinogen, and the resulting paste was more likely to break or crack, thus affecting the integrity and efficacy of the paste.

[0356] Table 1: Testing the ability of different compositions to be deployed from the syringe.

[0357]

[0358] a The control contained no gelatin and 35 mg of fibrinogen.

[0359] Conclusions

[0360] The results unexpectedly demonstrated that the paste compositions according to the present disclosure remained deployable from the syringe for at least 90 minutes after mixing. Thus, paste compositions comprising a biocompatible polymer (such as gelatin), thrombin, and fibrinogen can be prepared in a deployable paste form and maintain that form for a period compatible with clinical use during surgery.

[0361] Example 2: Efficacy testing of gelatin-thrombin pastes containing different amounts of fibrinogen

[0362] Objectives

[0363] To study the hemostatic efficacy of the paste composition of Example 1, i.e., a paste composition comprising crosslinked gelatin, thrombin, and fibrinogen, compared to a paste composition containing only gelatin and thrombin. The hemostatic efficacy was tested in a porcine spleen biopsy model as described below.

[0364] Materials and methods

[0365] Experimental model: The porcine spleen biopsy-drilling model was used, creating an 8-mm puncture (3 mm deep) in the spleen with a 5-second initial pressure period, followed by a 120-second evaluation period and a 5-second subsequent pressure period.

[0366] The porcine spleen biopsy-drilling model is a recognized model for evaluating the in vivo hemostatic efficacy of hemostatic pastes (Hutchinson et al., 2015, Surgical Technology International XXVII). The porcine spleen biopsy-drilling model studied in the present invention is similar to the model used by Hutchinson et al., 2015.

[0367] Experimental animals: Pigs were the animals selected for this model because they have a large volume of blood (70 ml / kg) and a large-vessel spleen, enabling many hemostasis comparisons to be made in a single animal.

[0368] Sample preparation: The paste was prepared as described in Example 1. Three different amounts of fibrinogen were studied: 7, 35, and 70 mg. As a control, a paste without added fibrinogen was used. Except for the difference in fibrinogen content, the chemical and water contents in the test samples and the control were the same.

[0369] Surgical procedure: A midline abdominal incision was made to expose the spleen. An 8-mm drill hole (3 mm deep) was created in the spleen. The bleeding intensity was evaluated on a scale of 0-5 as described below. Only bleeding intensities of 3 and 4 were considered acceptable. The drill holes in the control sample or test samples were then ready. A new drill hole was created in each test sample. Each sample type was tested 7 times (n = 7). The samples were tested in a random order.

[0370] At the start and end of the test period, a 12-minute negative control using only a wet gauze was performed on each pig. The negative control served as an indication of the animal's bleeding ability throughout the study.

[0371] The main test parameter was the measurement of the time to haemostasis (TTH). TTH was defined as the total time minus the last haemostasis evaluation period to ensure that bleeding no longer occurred (i.e., no rebleeding).

[0372] The evaluation of bleeding intensity and the application of test samples and negative controls are described in detail below.

[0373] Bleeding intensity: The bleeding intensity of each drill hole was evaluated by a surgeon on a scale of 0-5 (see the table below). For each drill hole, the bleeding intensity was noted at t = 0. Only the tests on wounds with a bleeding intensity of 3-4 were used for further analysis.

[0374] Table: Hemorrhage intensity levels

[0375]

[0376] Negative control: Place a wet gauze directly on the drill hole. Apply digital pressure for 30 seconds, followed by a 120 - second hemostasis assessment period. Assess hemostasis (defined as no blood oozing from under the test article for 30 seconds). If hemostasis is not achieved within 120 seconds, apply an additional 30 seconds of digital pressure and re - assess hemostasis for 120 seconds. Apply tamponade and observe until bleeding stops and hemostasis is achieved, or until the test period reaches 12 minutes. For the negative control, hemostasis was not achieved within the 12 - minute test period, thus demonstrating the pig's ability to bleed throughout the study period.

[0377] Administration of the test sample: Apply approximately 1 - 2 mL of the paste directly into the drill hole using the applicator tip. During application, the tip penetrates into the drill hole to ensure tissue contact. After application, place a gauze moistened with 0.9% saline on the drill hole. Apply digital pressure (tamponade) for 5 seconds. Stop pressing and remove the gauze, then assess hemostasis. If no blood is found oozing from under the test article for 120 seconds, conclude that hemostasis has been achieved and end the experiment. If blood oozes from under the test article within the 120 - second time frame, record the bleeding time and re - apply digital pressure for 5 seconds, then test for hemostasis. Continue this procedure until hemostasis is achieved or until 12 minutes have elapsed, whichever comes first.

[0378] Calculation example for assessing the hemostasis time: Apply digital pressure for 5 seconds, test for hemostasis: bleeding occurs after 39 seconds, re - apply digital pressure for 5 seconds, test for hemostasis for 120 seconds: no bleeding, conclusion: hemostasis is achieved after 5 + 39 + 5 seconds = 49 seconds, i.e., the last observation period is not included when calculating TTH.

[0379] Results

[0380] The results are shown in Table 2. The results demonstrate that, compared with 7 mg fibrinogen or the control (gelatin paste with thrombin but without fibrinogen), shorter mean hemostasis times were achieved using samples containing 70 and 35 mg fibrinogen.

[0381] Table 2: TTH of gelatin - thrombin paste compositions + / - fibrinogen

[0382]

[0383] a Values are shown as mean ± standard deviation. Each sample was tested 7 times (n = 7) in a random order.

[0384] Conclusion

[0385] Pastes containing gelatin, thrombin and fibrinogen cause hemostasis faster and more consistently than control pastes that do not have fibrinogen. Thus, the studies of the present invention demonstrate the benefit of including fibrinogen in hemostatic paste compositions that contain gelatin and thrombin.

[0386] Example 3: Efficacy Testing of Gelatin-Thrombin Paste Containing Fibrinogen

[0387] Objective

[0388] To investigate the hemostatic efficacy of the compositions according to the present disclosure.

[0389] Materials

[0390] 1 g of dry cross-linked gelatin particles, 2000 IU of thrombin, and 70 or 105 mg of fibrinogen in a 10 mL syringe (first syringe).

[0391] 6 mL of aqueous solution in a 10 mL syringe (second syringe).

[0392] The gelatin particles were obtained from ground cross-linked gelatin sponges (Spongostan), and the chemical and water contents in the test samples and the control were the same except for the difference in fibrinogen content.

[0393] Methods

[0394] The two syringes were interconnected via a Luer lock, and 6 mL of the aqueous solution was transferred into the syringe containing the dry powder composition. The resulting mixture was transferred back and forth between the first syringe and the second syringe until mixed, yielding a hemostatic composition contained in the second syringe.

[0395] The same porcine spleen biopsy-borehole model as described in Example 2 was used. Each sample type was tested 11 times (n = 11). The samples were tested in a random order.

[0396] Results

[0397] The results of the reconstituted hemostatic pastes are shown in Table 3. The results demonstrate that a shorter mean hemostasis time was achieved using the reconstituted paste with fibrinogen compared to the control.

[0398] Table 3: TTH of Gelatin-Thrombin Paste Compositions + / - Fibrinogen

[0399]

[0400] a Values are shown as mean ± standard deviation. Each sample was tested 11 times (n = 11) in a random order.

[0401] Compared with the sample containing 70 mg of fibrinogen, the force required to deploy the sample with 105 mg of fibrinogen is significantly increased, and the resulting paste is more likely to break or crack, thus affecting the integrity and efficacy of the paste.

[0402] Conclusion

[0403] The hemostatic composition according to the present invention causes hemostasis faster and more consistently than a control paste without fibrinogen.

Claims

1. A hemostatic composition comprising: a) one or more biocompatible polymers in particulate form, b) fibrinogen in an amount of 1 to 150 mg per gram of biocompatible polymer, and c) thrombin in an amount of 100 to 5000 IU per gram of biocompatible polymer.

2. The hemostatic composition according to claim 1, wherein the one or more biocompatible polymers in particulate form are present in an amount of 80% to 99% by weight of the composition, such as 81% to 99% by weight of the composition, such as 82% to 99% by weight of the composition, such as 83% to 99% by weight of the composition, such as 84% to 99% by weight of the composition, such as 85% to 99% by weight of the composition, such as 86% to 99% by weight of the composition, such as 87% to 99% by weight of the composition, such as 88% to 99% by weight of the composition, such as 89% to 99% by weight of the composition, such as 90% to 99% by weight of the composition.

3. The hemostatic composition according to any one of claims 1 to 2, wherein the one or more biocompatible polymers in particulate form are present in an amount of 0.05 to 20 g, such as 0.2 to 10 g, such as 0.5 to 2 g, such as about 1 g.

4. The hemostatic composition according to any one of the preceding claims, wherein the one or more biocompatible polymers in particulate form comprise a biocompatible polymer selected from the group consisting of or consist of a biocompatible polymer selected from the group consisting of consisting of: gelatin, collagen, chitin, chitosan, alginate, cellulose, oxidized cellulose, carboxymethyl cellulose, polyglycolic acid, polyacetic acid, and combinations thereof.

5. The hemostatic composition according to any one of the preceding claims, wherein the one or more biocompatible polymers comprise or consist of powder particles that are substantially insoluble in an aqueous medium.

6. The hemostatic composition according to any one of the preceding claims, wherein the one or more biocompatible polymers are crosslinked.

7. The hemostatic composition according to any one of the preceding claims, wherein the one or more biocompatible polymers are bioabsorbable.

8. The hemostatic composition according to any one of the preceding claims, wherein the one or more biocompatible polymers in particulate form comprise or consist of crosslinked gelatin particles.

9. The hemostatic composition according to claim 8, wherein the gelatin is obtained from micronized gelatin sponge or hydrogel.

10. The hemostatic composition according to any one of claims 1 to 9, wherein the biocompatible polymer particles have an average size of 1 to 1000 μm, such as 100 to 800 μm, such as 300 to 500 μm, such as about 450 microns, as measured by laser diffraction.

11. The hemostatic composition according to any one of the preceding claims, wherein the fibrinogen is human fibrinogen.

12. The hemostatic composition according to any one of the preceding claims, wherein the fibrinogen is recombinant human fibrinogen.

13. The hemostatic composition according to any one of the preceding claims, wherein the composition comprises 5 to 100 mg of fibrinogen per gram of the biocompatible polymer, such as 20 to 80 mg per gram of the biocompatible polymer, such as about 35 mg or such as about 70 mg of fibrinogen.

14. The hemostatic composition according to any one of the preceding claims, wherein the composition comprises 20 to 100 mg of fibrinogen per gram of the biocompatible polymer, such as about 30 mg to about 100 mg.

15. The hemostatic composition according to any one of the preceding claims, wherein the composition comprises fibrinogen in an amount of 1% to 10% by weight, such as 2% to 5% by weight, such as about 3.5% by weight.

16. The hemostatic composition according to any one of the preceding claims, wherein the thrombin is human thrombin.

17. The hemostatic composition according to any one of the preceding claims, wherein the thrombin is recombinant human thrombin.

18. The hemostatic composition according to any one of the preceding claims, wherein the composition comprises 400 to 4000 IU of thrombin per gram of the biocompatible polymer, such as 300 to 3000 IU per gram of the biocompatible polymer, such as 500 to 2500 IU, such as about 2000 IU of thrombin.

19. The hemostatic composition according to any one of the preceding claims, wherein the composition comprises a ratio of thrombin to fibrinogen of 0.5 IU / mg to 5000 IU / mg, such as 1 IU / mg to 2000 IU / mg, such as 2 IU / mg to 150 IU / mg, such as about 10 to 100 IU / mg.

20. The hemostatic composition according to any one of the preceding claims, wherein the composition comprises: a) about 1 g of a biocompatible polymer in particulate form, such as gelatin, b) about 35 mg of fibrinogen, c) about 2000 IU of thrombin.

21. The hemostatic composition according to any one of the preceding claims, which further comprises one or more additional active ingredients capable of stimulating hemostasis, wound healing, bone healing, tissue healing, and / or tendon healing.

22. The hemostatic composition according to claim 21, wherein the one or more active ingredients are selected from the group consisting of factor XIII, tranexamic acid, bone morphogenetic protein, metalloproteinase, insulin-like growth factor 1 (IGF-I), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), basic fibroblast growth factor, transforming growth factor β (TGF-β), and flakes or granules of extracellular matrix (ECM).

23. The hemostatic composition according to any one of the preceding claims, which further comprises one or more hydrophilic compounds.

24. The hemostatic composition according to claim 23, wherein the one or more hydrophilic compounds comprise polyethylene glycol (PEG).

25. The hemostatic composition according to claim 23, wherein the one or more hydrophilic compounds are one or more polyols selected from sugar alcohols, sugars, and / or their derivatives.

26. The hemostatic composition according to claim 25, wherein the sugar alcohol is selected from the group consisting of ethylene glycol, glycerol, erythritol, threitol, arabitol, xylitol, ribitol, mannitol, sorbitol, dulcitol, fucitol, iditol, inositol, heptitol, isomaltulose, maltitol, lactitol, polyhydric sugar alcohols, and mixtures thereof.

27. The hemostatic composition according to any one of claims 23 to 26, wherein the hydrophilic compound is present in an amount of 0.01 g to 0.5 g of hydrophilic compound per gram of biocompatible polymer, such as 0.01 g to 0.4 g per gram of biocompatible polymer, such as 0.01 to 0.3 g, such as 0.01 to 0.2 g, such as 0.01 to 0.1 g, such as 0.01 to 0.05 g of hydrophilic compound.

28. The hemostatic composition according to any one of the preceding claims, further comprising one or more extrusion enhancers, such as albumin, preferably human serum albumin.

29. The hemostatic composition according to any one of the preceding claims, wherein the composition contains less than 10% water by weight, preferably less than 5% water by weight, preferably less than 1% water by weight.

30. The hemostatic composition according to any one of the preceding claims, wherein the composition is sterile.

31. A method for preparing a hemostatic composition, which comprises the following steps: a) providing a hemostatic composition according to any one of claims 1 to 30, and b) adding a certain amount of aqueous medium to the hemostatic composition of a).

32. The method according to claim 31, wherein the amount of the aqueous medium added is 2 to 12 mL per gram of the hemostatic composition provided in step a), such as 3 to 10 mL per gram of the hemostatic composition, such as 4 to 8 mL, such as 4 to 7 mL, such as 5 to 6 mL.

33. A hemostatic composition obtained by the method according to any one of claims 31 to 32.

34. A hemostatic composition obtained by the method according to any one of claims 31 to 32, wherein the hemostatic composition is a flowable composition.

35. The hemostatic composition according to any one of claims 33 to 34, wherein the composition remains flowable for at least 4 hours, such as at least 2 hours, such as at least 1 hour, such as at least 30 minutes after adding the aqueous medium.

36. The hemostatic composition according to any one of claims 33 to 35, wherein the composition can be deployed from a syringe for at least 4 hours, such as at least 2 hours, such as at least 1 hour, such as at least 30 minutes after adding the aqueous medium.

37. A hemostatic composition comprising: a) one or more biocompatible polymers in particulate form, in an amount of 7% to 34% by weight, b) fibrinogen, in an amount of 0.008% to 5% by weight, c) thrombin, in an amount of 7 to 1700 IU per gram of the hemostatic composition, and d) an aqueous medium.

38. The hemostatic composition according to claim 37, wherein the hemostatic composition is as defined in any one of claims 1 to 36.

39. The hemostatic composition according to any one of claims 37 to 38, wherein the biocompatible polymer is present in an amount of 10% to 30% by weight, such as 10% to 25% by weight, such as 10% to 20% by weight, such as about 15% by weight.

40. The hemostatic composition according to any one of claims 37 to 39, wherein the fibrinogen is present in an amount of 0.01% to 2.5% by weight, such as 0.1% to 1.5% by weight, such as 0.3% to 1.2% by weight.

41. The hemostatic composition according to any one of claims 37 to 40, wherein the fibrinogen is present in an amount of 0.5% to 1.2% by weight.

42. The hemostatic composition according to any one of claims 37 to 41, wherein the thrombin is present in an amount of 16 to 900 IU per gram of the hemostatic composition, such as 150 to 700 IU per gram of the hemostatic composition.

43. The hemostatic composition according to any one of claims 37 to 42, wherein the thrombin is present in an amount of 280 to 350 IU per gram of the hemostatic composition.

44. The hemostatic composition according to any one of claims 37 to 43, wherein the hydrophilic compound is present in an amount of 1% to 20% by weight, such as 1% to 15% by weight, such as 1% to 10% by weight.

45. The hemostatic composition according to any one of claims 37 to 44, wherein the aqueous medium is selected from the group consisting of water, saline, calcium chloride solution, and buffered aqueous medium.

46. The hemostatic composition according to any one of claims 37 to 45, wherein the aqueous medium contains calcium ions.

47. The hemostatic composition according to any one of claims 37 to 46, wherein the composition is sterile.

48. The hemostatic composition according to any one of claims 37 to 47, wherein the composition is a flowable composition.

49. The hemostatic composition according to any one of claims 37 to 48, wherein the composition is a paste.

50. The hemostatic composition according to any one of claims 1 to 30 or 33 to 49, which is used to promote hemostasis and / or wound, bone, tendon, and / or tissue healing in an individual in need.

51. A container, which contains the hemostatic composition according to any one of claims 1 to 30 or the hemostatic composition according to any one of claims 33 to 49.

52. The container according to claim 51, wherein the container is an applicator, such as a syringe.

53. A kit, which contains: a) a first container, the first container containing the composition according to any one of claims 1 to 30, b) a second container, the second container containing an aqueous medium; and c) an optional outer package, wherein the two containers are interconnected.

54. The kit according to claim 53, wherein the first container and / or the second container is a syringe.

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