JP2020502241A - Hemostatic composition containing anion exchanger and calcium salt - Google Patents

Abstract

Provided are compositions comprising an anion exchanger and a calcium salt, and methods for preparing the same, for use as an agent for effecting hemostasis at a bleeding site.

Description

  The present invention relates to the field of hemostatic compositions. More specifically, the present invention relates to a hemostatic composition comprising an anion exchanger and calcium, and a method of using the same.

  Bleeding is a term commonly used to describe the leakage of blood from the vertebrate circulatory system. Bleeding can occur inside the body (internal bleeding) or outside the body (external bleeding). The site of bleeding can be almost any area of the body. Typically, internal bleeding occurs when blood leaks due to damage to blood vessels or organs. External bleeding occurs either when blood exits through skin damage or when blood exits through natural openings in the body, such as the mouth, nose, ears, vagina, or rectum.

  Bleeding can be traumatic injury (including abrasions, scratches, lacerations, incisions, puncture wounds with tools such as needles or knives, crush wounds, and gunshots), or related clotting disorders that are deficient in clotting factors It can be caused by a wide variety of disorders or conditions, including certain medical conditions, such as having a subject. In addition, bleeding may be caused by some non-steroidal anti-inflammatory drugs (NSAIDs) or anticoagulants, such as warfarin, low molecular weight heparin, apixaban (ELIQUIS®), dabigatran (PRADAXA) (Registered trademark)), edoxaban (SAVAYSA®), and) rivaroxaban (XARELTO®).

  Ongoing, untreated bleeding can result in phlebotomy, ie, excessive loss of blood volume (hypovolemia), and can lead to death.

  Stopping or controlling bleeding is called hemostasis, which involves blood clotting, and promoting, accelerating, or enhancing this mechanism is an important part of both first aid and surgery. Agents and compositions that enhance, promote, or accelerate hemostasis are referred to as "hemostatic agents."

  The hemostatic agent may include a hemostat, a sealant, or an adhesive. Typically, hemostatic agents include mechanical hemostatic agents (such as gelatin, collagen, oxidized regenerated cellulose), active hemostatic agents (such as thrombin), flowable hemostatic agents (such as a gelatin matrix used in combination with thrombin), and fibrin sealants. Subdivided.

  Some known hemostats require long preparation times before use, with a number of steps wasting valuable time in an emergency.

  Although biological hemostats are very effective, they have potential safety risks and their production costs are high.

  Many known hemostats require refrigeration, which is not available in developing countries and in various situations (including on the battlefield, in isolated areas, in emergencies), or during electrical hazards Or can be very expensive. In addition, refrigeration requirements increase manufacturing, shipping, and storage costs.

  Many known hemostatic agents are not effective for patients using anticoagulants such as heparin, aspirin, or coumadin.

  Many known hemostatic agents include blood clotting agents with high production costs, short shelf life, and usually requiring refrigeration, with the potential risk of contamination.

  Therefore, there is a need for a safe and effective hemostat that does not have at least some of the disadvantages of the prior art.

  Background art includes U.S. Patent Nos. 5,502,042 and 8,741,335, U.S. Patent Application Publication No. 2009/0062849, and WO 2013/071235 and WO 1993/05822. .

  The present invention provides a hemostatic composition comprising an anion exchanger, calcium, and a pharmaceutically acceptable carrier, and methods of using the same to provide hemostasis.

  According to one aspect of some embodiments of the present invention, there is provided a method of providing hemostasis at a bleeding site in a subject in need thereof, comprising the step of bleeding an effective amount of a hemostatic composition comprising an anion exchanger and a calcium salt. A method is provided that includes applying to a site.

  According to some embodiments, the anion exchanger comprises one or more positively charged groups attached to the matrix.

  According to some embodiments, the positively charged group (also called a polycation) is provided by a base selected from the group consisting of a strong base, a weak base, and combinations thereof.

  According to some embodiments, the strong base comprises a quaternary amino group.

  According to some embodiments, the weak base comprises an amino group selected from the group consisting of a primary amino group, a secondary amino group, a tertiary amino group, and combinations thereof.

  According to some embodiments, the weak base comprises a diethylaminoethyl (DEAE) group.

  According to some embodiments, the matrix is selected from the group consisting of aliphatic polyesters, polysaccharides, polypeptides, polystyrene-divinylbenzene, proteins (such as collagen gelatin or albumin), silica, and combinations thereof. .

  According to some embodiments, the matrix is crosslinked and optionally covalently crosslinked.

  According to some embodiments, the composition is substantially devoid of any protein of the blood clotting cascade.

  According to some embodiments, the composition is in a form selected from the group consisting of a slurry, a powder, a fiber, a film, a patch, and a liquid.

  According to some embodiments, the composition in slurry or liquid form further comprises a pharmaceutically acceptable carrier.

  According to some embodiments, the application is performed by applying pressure to the composition, if desired, toward the site of bleeding.

  According to an aspect of some embodiments of the present invention there is provided a hemostatic composition comprising an anion exchanger, a calcium salt, and optionally a pharmaceutically acceptable carrier.

  According to some embodiments, the anion exchanger comprises one or more positively charged groups attached to the matrix.

  According to some embodiments, the anion exchanger is linked to a solid phase.

  According to some embodiments, the positively charged group comprises a base selected from the group consisting of a strong base, a weak base, and combinations thereof.

  According to some embodiments, the strong base comprises a quaternary amino group.

  According to some embodiments, the weak base comprises an amino group selected from the group consisting of a primary amino group, a secondary amino group, a tertiary amino group, and combinations thereof.

  According to some embodiments, the weak base comprises a diethylaminoethyl (DEAE) group.

  According to some embodiments, the matrix is selected from the group consisting of aliphatic polyesters, polysaccharides, polypeptides, polystyrene-divinylbenzene, silica, and combinations thereof.

  According to some embodiments, the matrix is crosslinked and optionally covalently crosslinked.

  According to some embodiments, the polysaccharide is selected from the group consisting of cellulose, dextran, agarose, and combinations thereof.

  According to some embodiments, the protein is a structural protein, such as collagen or gelatin, or a protein that is abundant in plasma, such as albumin.

  According to some embodiments, the composition is substantially devoid of any protein of the blood clotting cascade.

  According to some embodiments, the composition is in a form selected from the group consisting of a slurry, a powder, a film, a patch, and a liquid.

  According to some embodiments, the composition in slurry or liquid form further comprises a pharmaceutically acceptable carrier.

  According to an aspect of some embodiments of the present invention there is provided a hemostatic composition comprising diethylaminoethyl (DEAE) and a calcium salt bound to a matrix.

  According to an aspect of some embodiments of the present invention, preparing an anion exchanger by covalently attaching one or more positively charged groups to a cross-linked matrix, and adding a calcium salt to the anion exchanger. And a method for preparing a hemostatic composition, the method comprising:

  According to an aspect of some embodiments of the present invention there is provided a hemostatic composition obtainable by a method disclosed herein.

  As used herein, the term "providing hemostasis" refers to causing, causing, promoting, accelerating, and / or enhancing hemostasis.

  As used herein, the term "bleeding site" refers to a site that is actively bleeding and that has bleeding complications, such as, for example, surgical sites, anastomotic sites, and / or suture sites. Refers to sites that may be susceptible to or tend to be affected.

  As used herein, the term "pharmaceutically acceptable carrier" refers to any inert diluent or vehicle that has no biological activity and is suitable for use in humans or other animals. Point. Carriers are phosphate buffered solution (PBS), physiological saline, sodium chloride solution, calcium chloride solution, lactated ringers (LR), 5% dextrose solution in normal physiological saline, various saccharides , Sugar alcohols (such as mannitol, sorbitol), and water for injection, such as, but not limited to, any carrier known in the art.

  As used herein, the term "slurry" refers to a thick, soft, wet material. Typically, a slurry is produced by mixing a dry component (eg, a powder or solid hydrophilic particles) with a liquid. The dry component concentration can be from 0.5% to 99% w / w of the total slurry composition. Typically, the slurry is a material that can be formed in a temperature range of 15 to 40C.

  As used herein, the term "lacking" with respect to components of a composition refers to components that are present in the composition at a concentration of less than 0.1% w / w of the total composition.

  As used herein, “comprising,” “including,” “having,” and grammatical variations thereof, refer to the recited feature, integer, step, or configuration. It will be understood that elements are specified, but does not exclude the addition of one or more additional features, integers, steps, components, or groups thereof. These terms encompass the terms “consisting of” and “consisting essentially of”.

  As used herein, the indefinite articles "one (a)" and "one (an)" mean "at least one" or "one" unless the context clearly indicates otherwise. One or more ".

  As used herein, the term “about” refers to ± 10%.

  Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition, the expressions, materials, methods, and examples are illustrative only and not intended to be limiting. In practicing the present invention, any methods and materials similar or equivalent to those described herein can be used.

  Some embodiments of the present invention are described herein with reference to the accompanying drawings. After reading the description with the drawings, it will become apparent to one skilled in the art how certain embodiments of the invention can be implemented. The drawings are for the purpose of exemplary discussion and are not intended to show structural details in the embodiments in any detail more than is necessary for a basic understanding of the present invention. For clarity, some objects shown in the figures are not to scale.

In the figure:
With a compression time of 30 seconds (for DEAE SEPHADEX ™ A-50) or 60 seconds (for commercial gelatin hemostat), DEAE SEPHADEX ™ A-50 (10% w / v) and commercial gelatin hemostasis Figure 4 shows the reduction of bleeding in an in vivo heparinized porcine spleen circular punch model (4 mm diameter, 2 mm depth) after application of the agent. Figure 3 shows a device used in an intractable bleeding model of pig spleen to create a "bullet-like" wound. Fig. 2b shows a wound made by the device of Fig. 2a in a pig spleen. As seen in FIG. 2b, the wound causes severe bleeding from the spleen. DEAE SEPHADEX ™ A-50 was prepared as a 10% w / v slurry in 20 mM CaCl ₂ and applied to the wound. Following application of the slurry, following a 4 minute compression time, the post-application bleeding intensity was evaluated as described in FIG. FIG. 9 shows a bullet-like wound after application of DEAE SEPHADEX ™ A-50 and tamponade, achieving complete hemostasis of a severely bleeding wound.

  The present invention provides a hemostatic composition comprising an anion exchanger, a calcium salt, and optionally a pharmaceutically acceptable carrier, and methods of using the same to achieve hemostasis.

  The principles, uses, and implementations of the teachings herein may be better understood with reference to the accompanying description. Upon review of this specification, one skilled in the art will be able to practice the invention without undue effort or experimentation.

  Before describing at least one embodiment in detail, it is to be understood that this invention is not necessarily limited in its application to the structure or arrangement of components and / or the details of the methods set forth in the following description. The invention is capable of other embodiments or of being practiced or carried out in various ways.

  The terms and terms used herein are for convenience and should not be construed as limiting.

  According to one aspect of some embodiments of the present invention, there is provided a method of providing hemostasis at a site of bleeding in a subject in need thereof, comprising the steps of: providing an effective amount of a hemostatic composition comprising an anion exchanger and a calcium salt. A method is provided that includes applying

  According to an aspect of some embodiments of the present invention there is provided a hemostatic composition comprising an anion exchanger and a calcium salt for use in providing hemostasis at a bleeding site.

  According to an aspect of some embodiments of the present invention there is provided the use of a hemostatic composition comprising an anion exchanger and a calcium salt in the manufacture of a medicament for providing hemostasis.

  According to an aspect of some embodiments of the present invention there is provided the use of a hemostatic pharmaceutical composition comprising an anion exchanger and a calcium salt in the manufacture of a medicament for providing hemostasis.

  According to some embodiments of the methods, compositions for use, use of the preparations, or methods disclosed herein, the anion exchanger comprises one or more positive ions attached to the matrix. It contains charged groups (at a pH in the range of 2-10) (also called polycations). In some such embodiments, the hemostatic composition lacks a polyanion.

  According to some embodiments of the method, pharmaceutical composition for use, use of the preparation, or method disclosed herein, the anion exchanger comprises one or more than one It contains positively charged groups (at a pH in the range of 2 to 10) (also called polycations). In some such embodiments, the hemostatic composition lacks a polyanion (such as a polyanionic polymer).

  Polyanions are molecules or chemical complexes with more than one negative charge. Polycations are molecules or chemical complexes with more than one positive charge.

  In one embodiment, the matrix may include anionic residues, but the overall net charge of the anion exchanger is positive.

  According to some embodiments, the positively charged groups are present in the hemostatic composition at a total ionic capacity of 2 mmol / g or more, for example, 2-5, 3-4 mmol / g.

  As used herein, the term "total ionic capacity" refers to the total amount of charged sites in the composition available for exchange. Total ionic capacity is expressed on a dry weight, wet weight, or wet volume basis.

  According to some embodiments, the anion exchanger is at a concentration of 0.5-99% w / v of the total hemostatic composition, optionally 5-15% w / v of the total hemostatic composition, e.g. Present at concentrations such as 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13% 14% or 15%.

  According to some embodiments, the anion exchanger is present at a concentration of at least 10% w / v of the total hemostatic composition, and the ratio between the anion exchanger and calcium in the hemostatic composition is: It is in the range of 1: 5 to 1:70. In some embodiments, this ratio is about 1:34.

  According to some embodiments, the positively charged group is selected from the group consisting of a strong base (such as one containing a quaternary amino group), a weak base (a primary amino group, a secondary amino group, a tertiary amino group). And a base selected from the group consisting of combinations thereof.

  According to some embodiments, the weak base comprises a diethylaminoethyl (DEAE) group.

  According to some embodiments, the positively charged group is attached to the matrix, eg, a matrix support, via a linker that is between the matrix support and the positively charged group.

  According to some embodiments, the matrix is an aliphatic polyester, polysaccharide, polypeptide (such as gelatin, bovine serum albumin (BSA), or collagen, or a combination thereof), polyacrylamide, acrylate copolymer, polystyrene-divinyl. It is selected from the group consisting of benzene, silica, and combinations thereof.

  According to some embodiments, the matrix is crosslinked and optionally covalently crosslinked. In some such embodiments, the matrix lacks ionic crosslinks.

  According to some embodiments, the polysaccharide is selected from the group consisting of cellulose, dextran, agarose, and combinations thereof.

  According to some embodiments, the matrix comprises SEPHADEX ™ (dextran), SEPHACEL ™ (cellulose), or TOYOPEARL ™ (hydroxylated methacrylic polymer), or a combination thereof. According to some embodiments, the composition is in a form selected from the group consisting of a slurry, a powder, a film, a patch, and a liquid. According to some such embodiments, the composition in slurry or liquid form further comprises a pharmaceutically acceptable carrier.

  According to some embodiments of the methods disclosed herein, applying the hemostatic composition to the bleeding site comprises applying pressure to the composition toward the bleeding site (eg, by gauze). Will be implemented.

  According to an aspect of some embodiments of the present invention there is provided a hemostatic composition comprising an anion exchanger, a calcium salt, and optionally a pharmaceutically acceptable carrier.

  According to some embodiments, the anion exchanger comprises one or more positively charged groups (also called polycations) attached to the matrix. In some such embodiments, the hemostatic composition is substantially devoid of a polyanion.

  According to some embodiments, the positively charged groups are present in the hemostatic composition at a total ionic capacity of 2 mmol / g or more, for example, 2-5, 3-4 mmol / g.

  According to some embodiments, the anion exchanger is present at a concentration of at least 10% w / v of the total hemostatic composition, and the ratio between the anion exchanger and calcium in the hemostatic composition is: It is in the range of 1: 5 to 1:70. In some embodiments, this ratio is about 1:34.

  According to some embodiments, the anion exchanger is at a concentration of 1-99% w / v of the total hemostatic composition, optionally 5-15% w / v of the total hemostatic composition, for example 5%. , 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% or 15%.

  According to some embodiments, the positively charged group comprises a strong base (such as one containing a quaternary amino group), a weak base (a primary amino group, a secondary amino group, a tertiary amino group, and combinations thereof). Such as those containing an amino group selected from the group), and bases selected from the group consisting of these combinations.

  According to some embodiments, the weak base comprises a diethylaminoethyl (DEAE) group.

  According to some embodiments, the positively charged group is attached to the matrix via a linker present between the matrix support and the positively charged group.

  According to some embodiments, the matrix is a polysaccharide, polypeptide (such as gelatin, bovine serum albumin (BSA), or collagen, or a combination thereof), polyacrylamide, acrylate copolymer, polystyrene-divinyl. It is selected from the group consisting of benzene, silica, and combinations thereof.

  According to some embodiments, the matrix is crosslinked and optionally covalently crosslinked. In some such embodiments, the matrix lacks ionic crosslinks.

  According to some embodiments, the polysaccharide is selected from the group consisting of cellulose, dextran, agarose, and combinations thereof.

  According to some embodiments, the matrix comprises SEPHADEX ™ (dextran), SEPHACEL ™ (cellulose), or TOYOPEARL ™ (hydroxylated methacrylic polymer), or a combination thereof.

  According to some embodiments, the composition is in a form selected from the group consisting of a slurry, a powder, a film, a patch, and a liquid. According to some such embodiments, the composition in slurry or liquid form further comprises a pharmaceutically acceptable carrier.

  According to some embodiments, the salts used herein are positive divalent cations.

  According to some embodiments of the methods, compositions for use, uses, or compositions for hemostasis disclosed herein, the calcium is calcium chloride, calcium acetate, calcium lactate, calcium oxalate, carbonate It is present in the hemostatic composition as a calcium salt, such as calcium, calcium gluconate, calcium phosphate, calcium glycerophosphate, or a combination thereof. In some embodiments, the calcium salt is calcium chloride, optionally present as a solution, and is optionally present at a concentration of 1-100 mM. In some such embodiments, the calcium present in the hemostatic composition is calcium chloride.

  According to some embodiments of the methods, compositions for use, uses, or compositions for hemostasis disclosed herein, the composition for hemostasis comprises substantially all proteins of the blood coagulation cascade. I lack.

  According to some embodiments, the matrix lacks the following polyanionic polymers: alginate and / or hyaluronate.

  According to some embodiments, the matrix comprises polystyrene sulfonate (such as sodium polystyrene sulfonate), polyacrylate (such as sodium polyacrylate), polymethacrylate (such as sodium polymethacrylate), polyvinyl sulfate (such as sodium polyvinyl sulfate), Polyphosphate (such as sodium polyphosphate), iota carrageenan, kappa carrageenan, gellan gum, carboxymethyl cellulose, carboxymethyl agarose, carboxymethyl dextran, carboxymethyl chitin, carboxymethyl chitosan, polymers modified with carboxymethyl groups, alginate (such as sodium alginate) ), A polymer comprising a plurality of carboxylate groups, xanthan gum, Beauty lack one or more crosslinkable polyanionic polymers are selected from the group consisting of.

  According to some embodiments, the polymer of the matrix has not been modified by the addition of carboxymethyl (CM) groups.

  According to some embodiments, the biocompatible polymer has been modified with diethylaminoethyl (DEAE) groups to obtain a cationic functional group, resulting in a polycationic polymer.

  According to some embodiments, the polycationic polymer is modified with chitosan (such as chitosan chloride), chitin, diethylaminoethyl-dextran, diethylaminoethyl-cellulose, diethylaminoethyl-agarose, diethylaminoethyl-alginate, diethylaminoethyl groups. Selected from the group consisting of polymers having a plurality of protonated amino groups, and polypeptides having an average residue isoelectric point of greater than 7, and combinations thereof. Preferably, the polycationic polymer is diethylaminoethyl-dextran (DEAE-dextran).

  According to some embodiments of the methods, compositions for use, uses, or compositions for hemostasis disclosed herein, the matrix lacks alginic acid and pectic acid.

  According to an aspect of some embodiments of the present invention there is provided a hemostatic composition comprising diethylaminoethyl (DEAE) and a calcium salt bound to a matrix.

  According to aspects of some embodiments of the present invention, preparing an anion exchanger by covalently attaching at least one positively charged group to a cross-linked matrix, and adding a calcium salt to the anion exchanger. And a method for preparing a hemostatic composition, comprising:

  According to a further aspect of such an embodiment of the present invention, there is provided a hemostatic composition obtainable by a method disclosed herein.

  According to some embodiments of the methods, compositions for use, uses, hemostatic compositions disclosed herein, the hemostatic composition substantially lacks all biological hemostatic agents. I.e., all protein components of the blood coagulation cascade, i.e., fibrinogen, fibrin, factor V, factor Va, factor VII, factor VIIa, factor VIII, factor VIIIa, factor IX, factor IXa , Factor X, factor Xa, factor XIa, factor XI, factor XII, factor XIIa, tissue factor (TF), and thrombin, and prothrombinase complex, prothrombin, and vWF, tannase complex Body, lacking high-molecular-weight kininogen (HMWK), prekallikrein, kallikrein, thromboplastin That.

  According to some embodiments of the methods, compositions for use, uses, or compositions for hemostasis disclosed herein, the composition for hemostasis comprises all proteins of the blood coagulation cascade (thrombin, prothrombin , And fibrinogen).

  In some embodiments, the anion exchanger comprises a matrix (also referred to as "support", "backing", "background", "base beads" or "resin"), which can be solid or semi-solid. The matrix may optionally be in the form of beads to which one or more positively charged groups are bound.

  Advantageously, the matrix can support, without disruption, the pressure typically applied during surgery when using auxiliary materials to stop bleeding.

  In some embodiments, the matrix comprises a cross-linked polymer. In some embodiments, the solid or semi-solid matrix material does not dissolve or disintegrate until hemostasis is achieved (at least 1 minute from application of the hemostatic composition).

  According to some embodiments, the polymer from which the matrix is formed is insoluble in water, preferably porous, and has an exclusion limit of at least 20 KDa.

  According to some embodiments, the matrix does not collapse when subjected to manual physical compression.

As used herein, the term “positively charged group” refers to ammonium, alkyl ammonium, dialkyl ammonium, trialkyl ammonium, quaternary ammonium, diethylaminoethyl (DEAE), dimethylaminoethyl (DMAE), triethyl aminoethyl, trimethyl aminoethyl, alkyl group, amino functional group _(e.g., NR ^{2 H} +), diethyl - (2-hydroxypropyl) aminoethyl, trimethylamino - hydroxypropyl, and at a pH range of a combination of these two. Refers to a molecule containing a chemical group that carries 0 to 10 positive charges.

  In one embodiment, the ion exchanger has multiple pKa values in the range of 6-14. In a further embodiment, the ion exchanger has a single pKa value greater than 9.

  According to some embodiments, the anion exchanger comprises DEAE bound to any matrix known to have hemostatic properties to enhance the hemostatic effectiveness of the matrix. Examples of suitable matrices include, but are not limited to, gelatin, cellulose, collagen, and starch.

  According to some embodiments, the hemostatic composition comprises at least a blend of an anion exchanger and calcium. The term "formulation" is intended to refer to any form of a homogeneous or heterogeneous mixture of at least an anion exchanger and calcium. The formulation may further comprise other components, if desired.

  According to some embodiments, the formulation is substantially free or lacks any protein component of the blood coagulation cascade (ie, such components comprise 0.1% of the total composition). present in the composition at a concentration of less than w / w), for example, the formulation may be substantially free of or lacking thrombin and fibrinogen.

  According to some embodiments, the formulation is a slurry, powder or liquid formulation.

  According to some embodiments, the formulation is provided in a frozen state, and prior to use, the product is thawed and brought to room temperature (i.e., 15-40 <0> C range) where the formulation is ready for use. To be.

  According to some embodiments, the compositions disclosed herein stop wound bleeding within one minute.

  As used herein, the term “hemostatic agent” or “hemostatic composition” refers to a material or composition that functions by causing blood to clot, ie, provide hemostasis. Typically, hemostatic agents increase blood clotting.

  In one embodiment, the term "providing hemostasis" in reference to a composition refers to a composition that causes the blood to clot by activation of a clotting factor such as prothrombin, resulting in cessation of bleeding or reduced bleeding intensity.

  As used herein, the term `` stop bleeding '' or `` stop bleeding '' with respect to the composition, when applied to a wound site, as described in the Materials and Methods section below, Refers to a composition that does not cause bleeding on a scale of, for example, 0 (no bleeding) to 5.

  As used herein, the term "reducing bleeding intensity" (also referred to herein as "hemostatic efficacy") refers to the difference between the initial bleeding intensity and the bleeding intensity after application.

  As used herein, the term "early bleeding intensity" refers to a morbidity immediately after wound formation and prior to application of the composition, e.g., from 0 to 5 as described in the Materials and Methods section below. Refers to the intensity of bleeding, as assessed on a scale.

  As used herein, the term `` bleeding intensity after application '' for a particular compression time, as described in the Materials and Methods section below, after application of the composition and after the compression time, Refers to the intensity of bleeding, for example, evaluated on a 0-5 scale.

  The hemostatic efficacy of the composition can be evaluated in terms of compression time when applied to a bleeding wound.

  As used herein, the term "compression time" refers to the time after application of the composition that manual compression is applied to the bleeding wound. Typically, this force is equal to the strength normally applied by the surgeon when using an auxiliary product to achieve hemostasis. In some embodiments where no compression is applied, the compression time is referred to as 0 seconds.

  In some embodiments, the compression time is about 8-12 minutes. In some embodiments, for intractable bleeding, the compression time is about 5 minutes. In some embodiments, for bleeding encountered in common surgical procedures, the compression time is about 1-2 minutes. "Intractable bleeding" is defined as class III bleeding and is defined above according to the World Health Organization classification. Typically, class III bleeding is associated with a 30-40% loss of circulating blood volume. Typical symptoms include decreased blood pressure, increased heart rate, and peripheral hypoperfusion (shock) in the patient.

  Without being bound by any theory, we hypothesize that thrombin is generated in situ when the compositions disclosed herein are applied to a bleeding wound or site. Surprisingly, it has been found that this in situ thrombin generation occurs to a degree and at a rate sufficient to achieve hemostasis.

  Advantageously, the presence of the physical matrix allows the hemostatic composition to be easily applied to the bleeding site, optionally by compression. Furthermore, the matrix itself, like oxidized regenerated cellulose (ORC), can contribute to coagulation by encapsulating platelets.

  As shown in the Examples section below, the matrix was found to be necessary for the hemostatic ability of positively charged functional groups such as DEAE. Typically, the matrix does not dissolve upon initial contact with the liquid and maintains its integrity until hemostasis is achieved, for example, for at least 1 minute, and aggregation of blood proteins may occur locally at the wound site. It should be of a nature that is concentrated and thus allows the initiation of the coagulation cascade. Advantageously, the matrix is stable under the pressure normally applied by surgeons during general surgery to achieve hemostasis. Advantageously, the nature of the matrix is such that it is distributed on and / or within the wound after application, optionally using compression.

  According to the present invention, it was found that various Sephadex types and commercial gelatin hemostats failed to stop bleeding (in a liver bleeding model), ie, no reduction in bleeding intensity was observed. .

  Various Sephadex types surprisingly reduced bleeding when containing the same base polymer, ie, cross-linked dextran provided as a powder to prepare a slurry.

  In addition, after application of DEAE Sephadex, the spleen was manually manipulated by folding the organ from both sides and no rebleeding occurred.

  Commercial gelatin hemostats failed to stop bleeding after a compression time of 60 seconds, whereas DEAE SEPHADEX ™ A-50 10% (w / v) was less effective after a compression time of 30 seconds. Was also found to successfully stop bleeding. It has also been found that compositions comprising an anion exchanger, such as DEAE, bound to a matrix, together with a calcium salt, provide complete hemostasis.

  These compositions provide substantially complete hemostasis regardless of the particular matrix used.

  Also, a composition comprising an anion exchanger such as DEAE bound to a matrix, such as SEPHADEX ™, SEPHACEL ™, and TOYOPEARL ™ (dextran, cellulose, and hydroxy methacrylic polymers, respectively) together with a calcium salt. The object has also been found to provide complete hemostasis.

More specifically, it was found that DEAE Sephadexin with CaCl ₂ was able to stop bleeding after 60 and 30 seconds of compression.

  It has been found that application of DEAE SEPHADEX ™ A-50 (8% w / v) can be used without compression to reduce bleeding intensity.

  The hemostatic ability of a composition comprising DEAE Sephadex (such as DEAE SEPHADEX ™ A-50) and a calcium salt may be determined, for example, by using commercially available thrombin when using the same compression time (such as 30 or 10 seconds compression). It has been found to exhibit the same efficacy as a gelatin hemostat containing. However, the hemostatic ability of hemostatic agents based on anion exchangers containing DEAE bound to the matrix and calcium salts is significantly better than that of commercially available gelatin hemostatic agents in the absence of biologically active ingredients such as thrombin. I was

  According to the present invention, it was found that compositions comprising DEAE Sephadex, prepared with NaCl but lacking calcium salts, did not reduce bleeding intensity.

  The results show that the use of a composition comprising a DEAE group attached to a matrix in the presence of a calcium salt achieved effective hemostasis.

  The results also showed that compression times of 30 and 60 seconds after application of DEAE Sephadex in the presence of the calcium salt resulted in complete hemostasis.

  The hemostatic time (TTH) of normal plasma in the presence of calcium (as measured, for example, by a coagulation assay) is about 200 seconds. On the other hand, the TTH of normal plasma in the presence of the calcium and anion exchanger according to the invention is in the range of about 10 to 180 seconds, for example in the range of about 10 to 60 seconds, in the range of about 10 to 30 seconds, about 15 60 seconds, about 15-30 seconds, and about 30-60 seconds. In one embodiment, TTH is about 30 seconds.

  According to the present invention, it was shown that anion exchangers such as DEAE bound to the matrix, together with calcium salts, provided complete hemostasis. This result was obtained regardless of the matrix used. The results are comparable to those obtained when using a commercially available hemostatic agent such as gelatin with thrombin.

  QAE SEPHADEX ™ was found to reduce bleeding with calcium salts.

  Compositions lacking calcium salts and / or matrices did not affect bleeding intensity.

  These results from DEAE and QAE suggest that compositions containing anion exchanger bound to the matrix and containing calcium salts are effective as hemostatic agents.

  The hemostatic ability of DEAE bound to a cross-linked polymer in the presence of a calcium salt was determined by Holcomb JB, Pusateri AE, Harris RA et al., Incorporated herein by reference as if fully set forth herein. Effect of dry fibrin sealant dressings versus gauze packing on blood loss in grade V liver injuries, which is difficult to solve, is disclosed by J.Traum. Model was further supported.

  DEAE bound to a crosslinked matrix according to the invention succeeded in achieving complete hemostasis in vivo in a heparinized porcine spleen circular punch model, while DEAE not bound to the matrix did not reduce bleeding intensity It was shown that.

  The density (ie, presence) of positively charged groups associated with the matrices disclosed herein has been shown to be important in achieving hemostasis. The charge on the matrix may advantageously be present at a density sufficient to achieve hemostasis according to the present invention.

  It has been found according to the present invention that not all positive charges evaluated provide the same level of hemostatic efficacy. Thus, in order to evaluate the charge density and the type of charge, an analysis can be performed to ensure that an optimal density range exists.

  For example, the synthesized matrices are monomerized by acid hydrolysis or the like, and applied to analytical instruments (eg, high pressure liquid chromatography, gas chromatography) capable of separating monomers based on various charges carried by them. can do. In this way, an analysis can be performed to evaluate the charge density on a particular molecule.

  Materials and methods

In vivo circular punch model.
The model is based on the model already described in WO 2012/087774 (A1), with some modifications. This model evaluates the efficacy of a test composition in reducing bleeding in vivo (haemostatic efficacy).

  First, the organ whose hemostasis is to be studied is exposed, then a single biopsy punch (4 mm in diameter, 2 mm in depth in Examples 1 and 5, 4 mm in diameter, 2 mm in depth in the second experiment of Example 2). 2 mm, or 8 mm in diameter, 3 mm in depth). The tissue in the punch was removed. The initial bleeding intensity ("early bleeding") is 0- "no bleeding", 1- "capillary bleeding", 2- "very mild bleeding", 3- "mild bleeding", 4- "moderate bleeding" Bleeding "and 5-" Severe bleeding "on a scale of 0-5.

  To assess the hemostatic efficacy of each test composition, approximately 0.5 ml of slurry or 100 mg of powder (for compositions applied as a powder) was applied to a hemorrhagic punch wound. The composition applied as a slurry was applied to the wound using a syringe, and the composition applied as a powder was applied directly on the wound.

  After application of the composition, manual compression was optionally applied using gauze for a specified period of time (also referred to herein as "compression time"). After compression, the gauze is removed and the bleeding intensity after application is immediately evaluated and after a further minute, qualitatively (yes / no) or quantitatively (using a scale of 0-5 as described above). )evaluated.

  Test compositions that reduce bleeding intensity (starting at least 3) to 1 (capillary bleeding) or 0 (no bleeding) were considered effective. For qualitative determination, the presence or absence of bleeding was visually examined, and gauze pieces pressed against the periphery of the treatment area were observed.

  The heparinized biopsy punch model is considered to be a suitable model for evaluating strong hemostasis.

  Time to Hemostasis (TTH) was evaluated after application of the composition. Time to hemostasis (TTH) was evaluated after application of the composition. TTH is defined as the time interval from application of the composition until complete hemostasis (score 0) is observed.

Example 1: Hemostatic properties of a composition comprising an anion exchanger and a calcium salt in an in vivo spleen model.
An initial assessment of the hemostatic properties of a composition comprising an anion exchanger and a calcium salt was performed using in vivo heparinized porcine spleen as described above using DEAE covalently attached to Sephadex (DEAE-SEPHADEX ™) as the anion exchanger. The test was performed with a circular punch model. In this experiment, the punch size was 4 mm in diameter and 2 mm in depth. After application, a compression time of 30 or 60 seconds was used. DEAE SEPHADEX ™ A-50 was tested at two concentrations. In this experiment, the bleeding intensity after application was qualitatively evaluated.

The following compositions (see details in Table 1 above) were evaluated for hemostatic efficacy:
1. DEAE SEPHADEX ™ A-50 prepared as a 10% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 50 mg DEAE SEPHADEX ™ A-50) (30 sec compression time ),
2. DEAE SEPHADEX ™ A-50 prepared as a 6.6% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 33 mg DEAE SEPHADEX ™ A-50) (60 seconds) Compression time),
3. SEPHADEX ™ G-75 Superfine prepared as a 10% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 50 mg of SEPHADEX ™ G-75 Superfine) (60 sec compression time ),
4. SEPHADEX ™ G-50 Medium prepared as a 10% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 50 mg of SEPHADEX ™ G-50 Medium) (60 sec compression time ),
5. Commercial gelatin hemostat prepared as a slurry (0.5 ml contains 55 mg of gelatin) (60 sec compression time).

  All four SEPHADEX ™ samples contain the same base polymer, cross-linked dextran. Compositions 1-4 were provided as powders for preparing slurries, and slurries were prepared as described in Table 1 above. A commercial gelatin flowable hemostat was used as a control.

  It was found that SEPHADEX (TM) G-50 Medium, SEPHADEX (TM) G-75 Superfine, and a commercially available gelatin hemostat could not stop bleeding, i.e., no decrease in bleeding intensity was observed. (Results not shown).

  Surprisingly, DEAE SEPHADEX ™ A-50 reduced bleeding at all compression times tested. Following application of DEAE SEPHADEX ™ A-50, the spleen was manually manipulated by folding the organ from both sides. No rebleeding occurred following any of the two different compression times at any of the concentrations tested (results not shown). Since hemostasis occurred only in the matrix supplemented with DEAE groups, it was concluded that the hemostatic effect was due to the presence of DEAE groups.

  FIG. 1 shows exemplary results obtained using DEAE SEPHADEX ™ A-50 10% (w / v) and commercially available gelatin. As shown in the figure, the commercial gelatin hemostat failed to stop bleeding after a compression time of 60 seconds, whereas DEAE SEPHADEX ™ A-50 10% (w / v) was 30%. Bleeding was successfully stopped even after a shorter compression time of seconds.

Example 2: Effect of a composition comprising an anion exchanger and calcium on hemostasis in an in vivo pig liver model.
In the following examples, the effects of each component of the composition comprising the anion exchanger and the calcium salt on hemostasis were evaluated separately and in combination using the in vivo heparinized porcine liver circular punch model as described above. . This experiment identifies which of the components of the composition are required to achieve hemostasis.

  The preparation of each composition is described in Table 1 above. The compression times are listed in Table 2 below. In this experiment, the initial bleeding intensity and the post-application bleeding intensity were evaluated according to a 0-5 scale.

The following compositions were evaluated:
1. DEAE SEPHADEX ™ A-50 prepared as an 8% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 40 mg of DEAE SEPHADEX ™ A-50),
2. A commercial gelatin hemostat prepared as a slurry (0.5 ml contains 55 mg of gelatin),
3. A commercially available gelatin hemostat containing thrombin prepared as a slurry (0.5 ml contains 55 mg of gelatin),
4. SEPHADEX ™ G-50 Medium prepared as a 14% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 70 mg of SEPHADEX ™ G-50 Medium),
5. DEAE SEPHADEX ™ A-50 prepared as an 8% w / v slurry in 20 mM NaCl solution (0.5 ml contains 40 mg DEAE SEPHADEX ™ A-50),
6. SP SEPHADEX ™ C-50 prepared as an 8% w / v slurry in a ₂₀ mM CaCl ₂ solution (0.5 ml contains 40 mg of SP SEPHADEX ™ C-50);
7. QAE SEPHADEX ™ prepared as an 8% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 40 mg QAE SEPHADEX ™),
8. 8. DEAE SEPHACEL ™ (100 mg) prepared as a slurry, and TOYOPEARL DEAE-650M (TM) (100 mg) prepared in powder form.

  Table 2 shows the results of compression time and bleeding intensity after application of each test composition. The bleeding intensity reduction was calculated by subtracting the post-application bleeding intensity from the initial bleeding intensity.

^＊初期出血強度から適用後の出血強度を減じることによって計算した。

^* Calculated by subtracting the post-application bleeding intensity from the initial bleeding intensity.

^＊初期出血強度から適用後の出血強度を減じることによって計算した。

^* Calculated by subtracting the post-application bleeding intensity from the initial bleeding intensity.

  In general, it can be seen that a composition comprising an anion exchanger such as DEAE bound to a matrix together with a calcium salt results in complete hemostasis (DEAE SEPHADEX ™ A-50, DEAE SEPHACEL ™, all containing calcium salts, And TOYOPEARL DEAE-650M ™ (see Table 2). These compositions provide substantially complete hemostasis regardless of the particular matrix used. For example, matrices such as SEPHADEX ™, SEPHACEL ™, and TOYOPEARL ™ (dextran, cellulose, and hydroxylated methacrylic polymers, respectively) had similar effects in reducing bleeding intensity.

More specifically, DEAE SEPHADEX ™ A-508 8% w / v in CaCl ₂ was able to stop bleeding after compression for 60 and 30 seconds. The results also showed that application of DEAE SEPHADEX ™ A-50 (8% w / v) could be used without compression to reduce bleeding intensity (Table 2).

  The hemostatic ability of a composition comprising DEAE SEPHADEX ™ A-50 and a calcium salt is demonstrated by the use of commercially available thrombin-containing gelatin hemostasis when using the same compression time (30 seconds) and even only 10 seconds of compression. It showed the same efficacy as the agent. However, the hemostatic ability of hemostatic agents based on anion exchangers containing DEAE bound to the matrix and calcium salts is significantly better than that of commercially available gelatin hemostatic agents in the absence of biologically active ingredients such as thrombin. I was

  As shown in Table 3, a composition prepared with NaCl and containing DEAE SEPHADEX ™ lacking a calcium salt did not result in a decrease in bleeding intensity, concluding that this sample was not effective in stopping bleeding be able to.

  In assessing the effect of ion exchange groups on hemostatic ability, it was shown that SP SEPHADEX ™, which contains an anionic group, sulfopropyl (SP) along with calcium salts, did not reduce bleeding intensity. In other words, materials having negative (SP) groups instead of positive (DEAE) groups were not effective as hemostatic agents.

  It was further shown that the quaternary aminoethyl with calcium salt, QAE SEPHADEX ™, was able to reduce bleeding intensity.

  Similar to Example 1, it was also shown that the matrix alone without functional groups (SEPHADEX ™ G-50 with calcium salt but no DEAE group) did not have hemostatic efficacy. .

  The results show that the use of a composition comprising a DEAE group attached to a matrix in the presence of a calcium salt achieved effective hemostasis.

  The results also show that 30 seconds and 60 seconds of compression after application of DEAE SEPHADEX ™ A-50 (8% w / v) resulted in complete hemostasis, thus defining TTH as 30 seconds. .

  Thus, it was shown that anion exchangers such as DEAE bound to the matrix, together with calcium salts, provided complete hemostasis. This result was obtained regardless of the matrix used. The results are comparable to those obtained when using commercially available hemostatic agents such as gelatin containing thrombin.

  QAE SEPHADEX ™ was found to reduce bleeding with calcium salts.

  Compositions lacking calcium salts and / or matrices did not affect bleeding intensity.

  These results suggest that a composition comprising an anion exchanger and a calcium salt is effective as a hemostat.

Example 3: Effect of a composition comprising an anion exchanger and a calcium salt on hemostasis in an in vivo porcine spleen model.
Previous experiments have shown that compositions comprising an anion exchanger consisting of DEAE conjugated to a crosslinked polymer and a calcium salt were effective in reducing bleeding intensity in an in vivo spleen circular punch model.

  In this experiment, the hemostatic activity of the composition was supported by another model, the in vivo heparinized porcine spleen circular punch model (performed as described above). This model was more severe in terms of bleeding intensity than the previous model. The compression time for all samples was 30 seconds. For samples 1-6, the punch size was 4 mm in diameter and 2 mm in depth. For samples 7-9, the punch size was 8 mm in diameter and 3 mm in depth. Typically, the severity of bleeding was increased by increasing punch size and / or adding heparin.

The following samples were tested for hemostatic efficacy:
1. DEAE SEPHADEX ™ A-50 prepared as an 8% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 40 mg of DEAE SEPHADEX ™ A-50),
2. DEAE SEPHADEX ™ A-50 prepared as a 10% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 50 mg DEAE SEPHADEX ™ A-50), tested in duplicate Done,
3. A commercial gelatin hemostat prepared as a slurry (0.5 ml contains 55 mg of gelatin),
4. A commercially available gelatin hemostat containing thrombin prepared as a slurry (0.5 ml contains 55 mg of gelatin),
5. SEPHADEX ™ G-50 Medium prepared as a 14% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 70 mg of SEPHADEX ™ G-50 Medium),
6. DEAE SEPHADEX ™ A-50 prepared as an 8% w / v slurry in 20 mM NaCl solution (0.5 ml contains 40 mg DEAE SEPHADEX ™ A-50),
7. Pure DEAE,
8. 8. CaCl ₂ (20 mM), and DEAE with CaCl _2.

DEAE SEPHADEX ™ A-508 8% and 10% w / v in the presence of CaCl ₂ can reduce bleeding intensity (approximately a 2-3 point decrease was observed), with a higher rate It was observed to give better results. In this model, the hemostatic ability of DEAE SEPHADEX ™ A-50 was superior to that of a commercial gelatin-based hemostatic agent with or without thrombin.

  As shown in liver experiments, both SEPHADEX ™ G-50 alone and DEAE SEPHADEX ™ with sodium salts did not reduce bleeding intensity.

  Pure DEAE did not reduce bleeding intensity. Therefore, it was concluded that when DEAE was not attached to the crosslinked polymer, it did not function as an effective hemostat (ie, no significant reduction in bleeding occurred). Addition of the calcium salt did not improve the hemostatic ability of the DEAE in the absence of the matrix, as seen in the composition comprising DEAE with the calcium salt composition (zero point reduction). It was also demonstrated that calcium salt alone did not have hemostatic ability.

  This experiment further supported the previous experiment and showed that a combination of calcium salts, DEAE groups, and matrix was required for demonstrated hemostatic efficacy.

Example 4: In vivo porcine spleen Hemostatic properties of a solution containing an anion exchanger and a calcium salt in a difficult bleeding model.
In this example, the hemostatic ability of DEAE bound to a cross-linked polymer in the presence of a calcium salt was determined by Holcomb JB, Pusateri AE, which is hereby incorporated by reference as if fully set forth herein. Harris RA, et al. (Effect of dry fibrin sealant dressings versus gauze packing on blood loss in grade V Liver is a solution from J. insures. It was further tested in a more difficult model of bleeding.

DEAE SEPHADEX ™ A-50 was prepared as a 10% w / v slurry in a 25 mM CaCl ₂ solution. The punch was performed using a dedicated device (see FIG. 2a), each arm was 5.5 cm long, the central hole was 1.5 cm deep, bullet shaped, and X 9.5 mm in diameter. This resulted in a W-shaped wound (FIG. 2b). Wounds represent bleeding that is difficult to resolve (such as a bullet injury).

  About 20 ml of the test composition was applied to the wound. Manual compression was applied to the wound site for 4 minutes. The tested composition stopped bleeding.

  The experiment was performed three times (two more repetitions) with similar results.

  In the first trial, if there was not enough material to achieve complete hemostasis, additional material was applied and compression was applied for another 3 minutes. Then complete hemostasis was achieved.

Example 5: Evaluation of Matrix Requirements In the following examples, the requirements of the matrix to which DEAE binds were further evaluated using the in vivo heparinized porcine spleen circular punch model as described above. The preparation of the composition is described in Table 1 above. The compression time was 60 seconds. In this experiment, the initial bleeding intensity and the post-application bleeding intensity were evaluated according to a 0-5 scale.

  DEAE Dextran 500 is a polycatonic derivative of dextran, prepared from dextran with an average molecular weight of 500 kD, with uncrosslinked dextran chains.

The following compositions were evaluated:
1. DEAE SEPHADEX ™ A-50 prepared as a 10% w / v slurry in ₂₀ mM CaCl ₂ solution (0.5 ml contains 40 mg DEAE SEPHADEX ™ A-50),
_2. DEAE dextran 500 prepared with 10% (w / w) CaCl ₂ powder (100 mg powder contained 90 mg DEAE dextran and 10 mg CaCl ₂ ).

  Table 4 shows the results of the bleeding intensity of various test compositions.

  DEAE bound to a cross-linked matrix succeeded in achieving complete hemostasis, while DEAE not bound to the matrix did not reduce bleeding intensity.

Example 6: Effect of positively charged groups on hemostasis The density (ie, the presence) of positively charged groups associated with the matrices disclosed herein has been shown to be important in achieving hemostasis. . The charge on the matrix should advantageously be present at a density sufficient to achieve hemostasis, as described above.

  Previous examples showed that not all positive charges evaluated provided the same level of hemostatic efficacy. Therefore, to evaluate the charge density and charge type, an analysis is performed to ensure that an optimal density range exists.

  For this purpose, the synthesized matrices are monomerized, such as by acid hydrolysis, and analytical instruments capable of separating monomers based on the various charges they carry (eg, high pressure liquid chromatography, gas chromatography). ). In this way, an analysis is performed to evaluate the charge density on a particular molecule.

  Although particular features of the invention have been described in the context of separate embodiments for clarity, it will be understood that they may also be presented in combination in a single embodiment. Would. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, but may also be implemented separately or in any suitable subcombination or It may suitably be provided in any of the other described embodiments of the invention. Certain features that are described in the context of various embodiments should not be considered as essential features of those embodiments unless the embodiment is inoperable without those elements. .

  While the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variances that fall within the scope of the appended claims.

  Citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention.

(Embodiment)
(1) A composition comprising an anion exchanger and a calcium salt for use as an agent for providing hemostasis at a bleeding site.
(2) The composition according to embodiment 1, wherein the anion exchanger comprises one or more positively charged groups attached to a matrix.
(3) The composition according to embodiment 2, wherein the positively charged group is provided by a base selected from the group consisting of a strong base, a weak base, and a combination thereof.
(4) The composition according to embodiment 3, wherein the strong base comprises a quaternary amino group.
(5) The composition according to embodiment 3, wherein the weak base includes an amino group selected from the group consisting of a primary amino group, a secondary amino group, a tertiary amino group, and a combination thereof.

(6) The composition according to embodiment 3, wherein the weak base comprises a diethylaminoethyl (DEAE) group.
(7) The composition according to any of embodiments 2 to 6, wherein said matrix is selected from the group consisting of aliphatic polyesters, polysaccharides, polypeptides, polystyrene-divinylbenzene, silica, and combinations thereof. .
(8) The composition according to any of embodiments 2 to 7, wherein the matrix is crosslinked.
(9) The composition according to embodiment 8, wherein the matrix is covalently crosslinked.
(10) The composition according to any of embodiments 1-9, wherein said composition is substantially devoid of any protein of the blood coagulation cascade.

(11) The composition according to any one of embodiments 1 to 10, wherein the composition is in a form selected from the group consisting of a slurry, a powder, a film, a patch, and a liquid.
(12) The composition of embodiment 11, wherein the composition in the form of a slurry or a liquid further comprises a pharmaceutically acceptable carrier.
(13) The composition according to any of embodiments 1-12, wherein said use comprises applying pressure to said composition, optionally towards said bleeding site.
(14) an anion exchanger;
Calcium salt,
A hemostatic composition, optionally comprising a pharmaceutically acceptable carrier.
(15) The composition for hemostasis according to embodiment 14, wherein the anion exchanger includes one or more positively charged groups bonded to a matrix.

(16) The hemostatic composition according to embodiment 15, wherein the positively charged group is provided by a base selected from the group consisting of a strong base, a weak base, and a combination thereof.
(17) The composition for hemostasis according to embodiment 16, wherein the strong base contains a quaternary amino group.
(18) The hemostatic composition according to embodiment 16, wherein the weak base comprises an amino group selected from the group consisting of a primary amino group, a secondary amino group, a tertiary amino group, and a combination thereof.
(19) The composition for hemostasis according to embodiment 16, wherein the weak base comprises a diethylaminoethyl (DEAE) group.
(20) The hemostatic agent according to any of embodiments 14 to 19, wherein the matrix is selected from the group consisting of aliphatic polyesters, polysaccharides, polypeptides, polystyrene-divinylbenzene, silica, and combinations thereof. Composition.

(21) The composition for hemostasis according to any of embodiments 15 to 20, wherein the matrix is crosslinked.
(22) The hemostatic composition according to embodiment 21, wherein the crosslinked polymer is covalently crosslinked.
(23) The hemostatic composition according to any of embodiments 20 to 22, wherein the polysaccharide is selected from the group consisting of cellulose, dextran, agarose, and combinations thereof.
(24) The composition for hemostasis according to any of embodiments 15 to 23, wherein the composition is substantially devoid of any protein of the blood coagulation cascade.
(25) The hemostatic composition according to any of embodiments 15 to 24, wherein the composition is in a form selected from the group consisting of a slurry, a powder, a film, a patch, and a liquid.

(26) The hemostatic composition according to embodiment 25, wherein the composition in the form of the slurry or liquid further comprises a pharmaceutically acceptable carrier.
(27) diethylaminoethyl (DEAE) bound to the matrix;
A hemostatic composition comprising a calcium salt.
(28) A method for preparing a hemostatic composition,
Preparing an anion exchanger by covalently attaching one or more positively charged groups to a cross-linked matrix;
Adding a calcium salt to the anion exchanger.
(29) A hemostatic composition obtainable by the method according to the embodiment 28.

Claims (28)

  A composition comprising an anion exchanger and a calcium salt for use as an agent for providing hemostasis at a bleeding site.   The composition of claim 1, wherein the anion exchanger comprises one or more positively charged groups attached to a matrix.   3. The composition of claim 2, wherein the positively charged group is provided by a base selected from the group consisting of a strong base, a weak base, and combinations thereof.   4. The composition according to claim 3, wherein said strong base comprises a quaternary amino group.   The composition of claim 3, wherein the weak base comprises an amino group selected from the group consisting of a primary amino group, a secondary amino group, a tertiary amino group, and combinations thereof.   The composition of claim 3, wherein said weak base comprises a diethylaminoethyl (DEAE) group.   The composition of any one of claims 2 to 6, wherein the matrix is selected from the group consisting of aliphatic polyesters, polysaccharides, polypeptides, polystyrene-divinylbenzene, silica, and combinations thereof.   The composition according to any one of claims 2 to 7, wherein the matrix is crosslinked.   9. The composition of claim 8, wherein said matrix is covalently crosslinked.   10. The composition according to any one of the preceding claims, wherein the composition is substantially devoid of any protein of the blood coagulation cascade.   The composition according to any one of claims 1 to 10, wherein the composition is in a form selected from the group consisting of a slurry, a powder, a film, a patch, and a liquid.   12. The composition according to claim 11, wherein the composition in the form of a slurry or a liquid further comprises a pharmaceutically acceptable carrier.   13. The composition according to any one of the preceding claims, wherein the use comprises applying pressure to the composition, optionally toward the bleeding site. An anion exchanger;
Calcium salt,
A hemostatic composition, optionally comprising a pharmaceutically acceptable carrier.   15. The hemostatic composition of claim 14, wherein the anion exchanger comprises one or more positively charged groups attached to a matrix.   The hemostatic composition according to claim 15, wherein the positively charged group is provided by a base selected from the group consisting of a strong base, a weak base, and a combination thereof.   17. The hemostatic composition according to claim 16, wherein the strong base contains a quaternary amino group.   17. The hemostatic composition according to claim 16, wherein the weak base includes an amino group selected from the group consisting of a primary amino group, a secondary amino group, a tertiary amino group, and a combination thereof.   17. The hemostatic composition according to claim 16, wherein the weak base comprises a diethylaminoethyl (DEAE) group.   The hemostatic composition according to any one of claims 14 to 19, wherein the matrix is selected from the group consisting of aliphatic polyesters, polysaccharides, polypeptides, polystyrene-divinylbenzene, silica, and combinations thereof. object.   The hemostatic composition according to any one of claims 15 to 20, wherein the matrix is cross-linked.   The hemostatic composition according to claim 21, wherein the crosslinked polymer is covalently crosslinked.   The hemostatic composition according to any one of claims 20 to 22, wherein the polysaccharide is selected from the group consisting of cellulose, dextran, agarose, and combinations thereof.   24. The hemostatic composition according to any one of claims 15 to 23, wherein the composition is substantially devoid of any protein of the blood coagulation cascade.   The hemostatic composition according to any one of claims 15 to 24, wherein the composition is in a form selected from the group consisting of a slurry, a powder, a film, a patch, and a liquid.   26. The hemostatic composition according to claim 25, wherein the composition in the form of a slurry or liquid further comprises a pharmaceutically acceptable carrier. Diethylaminoethyl (DEAE) bound to the matrix;
A hemostatic composition comprising a calcium salt. A method for preparing a hemostatic composition,
Preparing an anion exchanger by covalently attaching one or more positively charged groups to a cross-linked matrix;
Adding a calcium salt to the anion exchanger.

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