WO2019236932A1 - Coacervats complexes hémostatiques fluides - Google Patents

Coacervats complexes hémostatiques fluides Download PDF

Info

Publication number
WO2019236932A1
WO2019236932A1 PCT/US2019/035923 US2019035923W WO2019236932A1 WO 2019236932 A1 WO2019236932 A1 WO 2019236932A1 US 2019035923 W US2019035923 W US 2019035923W WO 2019236932 A1 WO2019236932 A1 WO 2019236932A1
Authority
WO
WIPO (PCT)
Prior art keywords
polyphosphate
hemostatic
complex coacervate
equal
chitosan
Prior art date
Application number
PCT/US2019/035923
Other languages
English (en)
Inventor
Russell Stewart
Monika SIMA
Original Assignee
University Of Utah Research Foundation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University Of Utah Research Foundation filed Critical University Of Utah Research Foundation
Publication of WO2019236932A1 publication Critical patent/WO2019236932A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
    • A61L24/08—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/0047—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L24/0073—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material with a macromolecular matrix
    • A61L24/0089—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material with a macromolecular matrix containing inorganic fillers not covered by groups A61L24/0078 or A61L24/0084
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/02—Surgical adhesives or cements; Adhesives for colostomy devices containing inorganic materials
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/04—Materials for stopping bleeding
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/06—Flowable or injectable implant compositions

Definitions

  • Bleeding can occur as a result of surgeries and other medical procedures, military combat, and injuries such as those sustained during traffic and transportation accidents, workplace and household accidents, participation in sporting events, by victims of violent crime, and the like. Uncontrolled bleeding can lead to the need for transfusions and/or additional surgeries for trauma patients and, in many cases, patient death. Reducing the duration and amount of bleeding leads to better patient outcomes and may also reduce costs associated with medical care.
  • Hemostasis is the physiological process that causes bleeding to stop from a wound in a subject.
  • Various hemostatic agents have historically been or are currently in use. However, many of these have drawbacks. Most hemostatic agents are dry powders or bandages. In the case when the hemostatic agent is a powder, it is limited to where agent can be applied, particularly where there is excessive blood flow or large volumes of blood.
  • hemostatic agents that are biocompatible, nontoxic, and biodegradable, but which are not hydrolyzed too quickly by the body’s enzymes to be effective for their intended purpose.
  • these hemostatic agents could be active at all stages of the hemostatic process, from initial vasoconstriction, through formation of a platelet plug to block any holes at the site of injury to a blood vessel, through recruitment of the remainder of the components of a clot.
  • the hemostatic agents would flow easily and would be injectable for example, through a syringe, cannula, or catheter, thus providing a minimally-invasive method to address internal bleeding.
  • these hemostatic agents would be active only where applied or at the site of injury and would not cause formation of platelet plugs or clots at non-injured sites.
  • the complex coacervates are composed of a poly cation and a poly anion.
  • the complex coacervates have properties that are superior when compared to traditional hemostatic agents.
  • “Optional” or“optionally” means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.
  • the phrase“optionally substituted lower alkyl” means that the lower alkyl group can or cannot be substituted and that the description includes both unsubstituted lower alkyl and lower alkyl where there is substitution.
  • alkyl group is a branched or unbranched saturated hydrocarbon group of 1 to 25 carbon atoms, such as methyl, ethyl, «-propyl, isopropyl, «-butyl, isobutyl, /-butyl, pentyl, hexyl, heptyl, octyl, decyl, tetradecyl, hexadecyl, eicosyl, tetracosyl and the like.
  • longer chain alkyl groups include, but are not limited to, a palmitate group.
  • A“lower alkyl” group is an alkyl group containing from one to six carbon atoms.
  • cycloalkyl group is a non-aromatic carbon-based ring composed of at least three carbon atoms.
  • examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.
  • heterocycloalkyl group is a cycloalkyl group as defined above where at least one of the carbon atoms of the ring is substituted with a heteroatom such as, but not limited to, nitrogen, oxygen, sulphur, or phosphorus.
  • aryl group as used herein is any carbon-based aromatic group including, but not limited to, benzene, naphthalene, etc.
  • the term“aryl group” also includes“heteroaryl group,” which is defined as an aromatic group that has at least one heteroatom incorporated within the ring of the aromatic group. Examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, and phosphorus. In one aspect, the heteroaryl group is imidazole. The aryl group can be substituted or unsubstituted.
  • the aryl group can be substituted with one or more groups including, but not limited to, alkyl, alkynyl, alkenyl, aryl, halide, nitro, amino, ester, ketone, aldehyde, hydroxy, carboxylic acid, or alkoxy.
  • inhibitor as used herein is defined as the cessation of bleeding in a subject when a hemostatic complex coacervate as described herein is administered to the wound.
  • the term“reduce” as used herein is the ability of the hemostatic complex coacervate as described herein to reduce the amount of bleeding in a subject when compared to the amount of bleeding from the same subject that is not treated with the hemostatic complex coacervate.
  • admixing refers to a combination of two or more components together when there is no chemical reaction or physical interaction between the components.
  • the terms“admixing” and“admixture” also includes the chemical interaction or physical interaction between any of the components described herein upon mixing to produce the composition.
  • the subject is a non-human animal (domesticated, wild, farm) such as, for example, a horse, cat, dog, cow, pig, sheep, goat, mouse, rabbit, chicken, rat, or guinea pig.
  • “Physiological conditions” refers to condition such as pH, temperature, etc. at a particular location within the subject.
  • Ranges may be expressed herein as from“about” one particular value, and/or to“about” another particular value. When such a range is expressed, another aspect includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent“about,” it will be understood that the particular value forms another aspect. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint.
  • references in the specification and concluding claims to molar amounts of a particular element or component in a composition or article denotes the molar relationship between the element or component and any other elements or components in the composition or article for which an amount is expressed.
  • X and Y are present at a molar ratio of 2:5, and are present in such ratio regardless of whether additional components are contained in the compound.
  • a weight percent of a component is based on the total weight of the formulation or composition in which the component is included.
  • each of the combinations A + E, A + F, B + D, B + E, B + F, C + D, C + E, and C + F is specifically contemplated and should be considered from disclosure of A, B, and C; D, E, and F; and the example combination of A + D.
  • any subset or combination of these is also specifically contemplated and disclosed.
  • the sub-group of A + E, B + F, and C + E is specifically contemplated and should be considered from disclosure of A, B, and C; D, E, and F; and the example combination of A + D.
  • Described herein are flowable hemostatic complex coacervates and applications thereof.
  • Polyelectrolytes such as a poly cation and a polyanion used herein with opposite net charges in aqueous solution can associate into several higher order morphologies depending on the solution conditions and charge ratios. They can form stable colloidal suspensions of polyelectrolyte complexes with net surface charges. Repulsion between like surface charges stabilize the suspension from further association.
  • the initial complexes When the polyelectrolyte charge ratios are balanced, or near balance, the initial complexes can further coalesce and settle out into a dense fluid phase in which the opposite macroion charges are approximately equal. This process is referred to as complex coacervation, and the dense fluid morphology as a complex coacervate.
  • the process is an associative macrophase separation of an aqueous solution of two oppositely charged poly electrolytes into two liquid phases, a dense concentrated polyelectrolyte phase in equilibrium with a poly electrolyte depleted phase.
  • the aqueous coacervate phase can be dispersed into the aqueous depleted phase but quickly settles back out, like oil droplets in water.
  • the spontaneous demixing of paired poly electrolytes into complex coacervates occurs when attractive forces between polyelectrolyte pairs are stronger than repulsive forces, yet the interactions are sufficiently dynamic to result in a liquid state.
  • gel is defined herein as non-fluid colloidal network or polymer network that is expanded throughout its whole volume by a fluid.
  • IUPAC Compendium of Chemical Terminology, 2nd ed. (the "Gold Book”). Compiled by A. D. McNaught and A. Wilkinson. Blackwell Scientific Publications, Oxford (1997).
  • the hemostatic liquid complex coacervates described herein are liquids (i.e., flowable).
  • the hemostatic complex coacervates described herein have a completely different morphology compared to corresponding gels despite the fact that the components in the hemostatic complex coacervates and the gel are identical.
  • Hemostasis is the process by which bleeding stops in the body. Hemostasis can be naturally achieved through the steps of (a) vasoconstriction to reduce blood flow to the site of injury, (b) formation of a platelet plug to temporarily block a break in the blood vessel wall, and (c) formation of a clot to seal the break until such time as tissues can heal. In emergency medicine situations, hemostasis can be aided by various externally applied agents including, but not limited to, chemical agents such as microfibrillar collagen, direct pressure (either by physically pressing or by applying a dressing to a wound), use of sutures, or use of physical agents such as, for example, gelatin sponges.
  • chemical agents such as microfibrillar collagen
  • direct pressure either by physically pressing or by applying a dressing to a wound
  • sutures or use of physical agents such as, for example, gelatin sponges.
  • hemostatic complex coacervates capable of reducing or inhibiting bleeding in a subject.
  • the hemostatic complex coacervates provided herein are effective at speeding the completion of one or more steps of hemostasis as outlined above.
  • the components used to produce the hemostatic complex coacervates described herein as well as their applications thereof are provided below.
  • the poly cation is generally composed of a polymer backbone with a plurality of cationic groups at a particular pH.
  • the cationic groups can be pendant to the polymer backbone and/or incorporated within the polymer backbone.
  • the poly cation is any biocompatible polymer possessing cationic groups or groups that can be readily converted to cationic groups by adjusting the pH.
  • the poly cation is a polyamine compound. The amino groups of the poly amine can be branched or part of the polymer backbone.
  • the amino group can be a primary, secondary, or tertiary amino group that can be protonated to produce a cationic ammonium group at a selected pH.
  • the polyamine is a polymer with a large excess of positive charges relative to negative charges at the relevant pH, as reflected in its isoelectric point (pi), which is the pH at which the polymer has a net neutral charge.
  • pi isoelectric point
  • the number of amino groups present on the poly cation ultimately determines the charge density of the poly cation at a particular pH.
  • the poly cation can have from 10 to 90 mole %, 10 to 80 mole %, 10 to 70 mole %, 10 to 60 mole %, 10 to 50 mole %, 10 to 40 mole %, 10 to 30 mole %, or 10 to 20 mole % amino groups.
  • the polyamine has excess positive charges at a pH of about 7, with a pi significantly greater than 7. As will be discussed below, additional amino groups can be incorporated into the polymer in order to increase the pi value.
  • the amino group can be derived from a residue of lysine, histidine, or arginine attached to the poly cation.
  • arginine has a guanidinyl group, where the guanidinyl group is a suitable amino group useful herein.
  • Any anionic counterions can be used in association with the cationic polymers.
  • the counterions should be physically and chemically compatible with the essential components of the composition and do not otherwise unduly impair product performance, stability or aesthetics.
  • Non-limiting examples of such counterions include halides (e.g., chloride, fluoride, bromide, iodide), sulfate, methylsulfate, acetate and other monovalent carboxylic acids.
  • the poly cation can be a positively-charged protein produced from a natural organism.
  • a recombinant P. californica protein can be used as the poly cation.
  • Pel, Pc2, Pc4-Pcl8 (SEQ ID NOS 1-17) can be used as the poly cation.
  • the type and number of amino acids present in the protein can vary in order to achieve the desired solution properties. For example, Pel is enriched with lysine (13.5 mole %) while Pc4 and Pc5 are enriched with histidine (12.6 and 11.3 mole %, respectively).
  • the poly cation is a recombinant protein produced by artificial expression of a gene or a modified gene or a composite gene containing parts from several genes in a heterologous host such as, for example, bacteria, yeast, cows, goats, tobacco, and the like.
  • the poly cation can be a biodegradable poly amine.
  • the biodegradable polyamine can be a synthetic polymer or naturally-occurring polymer.
  • the mechanism by which the poly amine can degrade will vary depending upon the poly amine that is used. In the case of natural polymers, they are biodegradable because there are enzymes that can hydrolyze the polymer chain. For example, proteases can hydrolyze natural proteins like gelatin. In the case of synthetic biodegradable polyamines, they also possess chemically labile bonds. For example, b-aminoesters have hydrolyzable ester groups.
  • other considerations such as the molecular weight of the poly amine and crosslink density of the hemostatic complex coacervate can be varied in order to modify the rate of biodegradability.
  • the biodegradable polyamine includes a polysaccharide, a protein, or a synthetic polyamine.
  • Polysaccharides bearing one or more amino groups can be used herein.
  • the polysaccharide is a natural polysaccharide such as chitosan or chemically modified chitosan.
  • the protein can be a synthetic or naturally-occurring compound.
  • the biodegradable polyamine is a synthetic polyamine such as poly(P-aminoesters), polyester amines, poly(disulfide amines), mixed poly (ester and amide amines), and peptide crosslinked poly amines.
  • the biodegradable polyamine can be an amine- modified natural polymer.
  • the amine-modified natural polymer can be gelatin modified with one or more alkylamino groups, heteroaryl groups, or an aromatic group substituted with one or more amino groups. Examples of alkylamino groups are depicted in Formulae IV-VI
  • R 13 -R 22 are, independently, hydrogen, an alkyl group, or a nitrogen containing substituent
  • s, t, u, v, w, and x are an integer from 1 to 10;
  • A is an integer from 1 to 50, where the alk lamino group is covalently attached to the natural polymer.
  • the natural polymer has a carboxyl group (e.g., acid or ester)
  • the carboxyl group can be reacted with an alkyldiamino compound to produce an amide bond and incorporate the alkylamino group into the polymer.
  • the amino group NR 13 is covalently attached to the carbonyl group of the natural polymer.
  • the number of amino groups can vary.
  • the alkylamino group is -NHCH 2 NH 2 , -NHCH 2 CH 2 NH 2 ,
  • the amine-modified natural polymer can include an aryl group having one or more amino groups directly or indirectly attached to the aromatic group.
  • the amino group can be incorporated in the aromatic ring.
  • the aromatic amino group is a pyrrole, an isopyrrole, a pyrazole, imidazole, a triazole, or an indole.
  • the aromatic amino group includes the isoimidazole group present in histidine.
  • the biodegradable polyamine can be gelatin modified with ethylenediamine.
  • the poly cation includes a polyacrylate having one or more pendant amino groups.
  • the backbone of the poly cation can be derived from the polymerization of acrylate monomers including, but not limited to, acrylates, methacrylates, acrylamides, and the like.
  • the poly cation backbone is derived from polyacrylamide.
  • the poly cation is a block co-polymer, where segments or portions of the co-polymer possess cationic groups or neutral groups depending upon the selection of the monomers used to produce the co polymer.
  • the poly cation can be a dendrimer.
  • the dendrimer can be a branched polymer, a multi-armed polymer, a star polymer, and the like.
  • the dendrimer is a polyalkylimine dendrimer, a mixed amino/ether dendrimer, a mixed amino/amide dendrimer, or an amino acid dendrimer.
  • the dendrimer is poly(amidoamine), or PAMAM.
  • the dendrimer has 3 to 20 arms, wherein each arm comprises an amino group.
  • the poly cation is a poly amino compound.
  • the polyamino compound has 10 to 90 mole % primary amino groups.
  • the poly cation polymer has at least one fragment of the formula I
  • R 1 , R 2 , and R 3 are, independently, hydrogen or an alkyl group
  • X is oxygen or NR 5 , where R 5 is hydrogen or an alkyl group
  • m is from 1 to 10, or the pharmaceutically-acceptable salt thereof.
  • R 1 , R 2 , and R 3 are methyl and m is 2.
  • the polymer backbone is composed of CH2-CR 1 units with pendant -C(0)X(CH 2 ) m NR 2 R 3 units.
  • the polycation is the free radical polymerization product of a cationic primary amine monomer (3-amino- propyl methacrylate) and acrylamide, where the molecular weight is from 10 to 200 kd and possesses primary monomer concentrations from 5 to 90 mol %.
  • the poly cation is a protamine.
  • Protamines are poly cationic, arginine-rich proteins that play a role in condensation of chromatin into the sperm head during spermatogenesis.
  • commercially available protamines purified from fish sperm, are readily available in large quantity and are relatively inexpensive.
  • a non-limiting example of a protamine useful herein is salmine.
  • the amino acid sequence of salmine, a protamine isolated from salmon sperm, is SEQ ID NO 18. Of the 32 amino acids, 21 are arginine (R).
  • the guanidinyl group on the sidechain of R has a pK a of -12.5, making salmine a densely charged poly cation at physiologically relevant pH. It has a molecular mass of -4,500 g/mol and a single negative charge at the carboxy terminus.
  • the protamine is clupein.
  • the poly cation is a natural polymer wherein one or more amine groups present on the natural polymer have been modified with a guanidine group.
  • the poly cation is a synthetic polymer containing one or more guanidinyl sidechains.
  • the poly cation can be a synthetic polyguanidinyl polymer having an acrylate or methacrylate backbone and one or more guanidinyl sidechains.
  • the poly cation polymer has at least one fragment of the formula VIII
  • R 1 is hydrogen or an alkyl group
  • X is oxygen or NR 5 , where R 5 is hydrogen or an alkyl group
  • m is from 1 to 10, or the pharmaceutically-acceptable salt thereof.
  • R 1 , R 2 , and R 3 are methyl and m is 2.
  • the polymer backbone is composed of CH2-CR 1 units with pendant -C(0)X(CH 2 ) m NC(NH)NH 2 units.
  • the synthetic polyguanidinyl polymer can be derivatized with one or more crosslinkable groups described herein.
  • one or more acrylate or methacrylate groups can be grafted onto the synthetic polyguanidinyl polymer.
  • the poly cationic component of the hemostatic complex coacervates described herein is chitosan.
  • Chitosan is an oligomer of b-(1 4)-linked d-glucosamine.
  • Chitosan can be prepared by the deacetylation and hydrolysis of chitin, which is commonly found in the exoskeletons of arthropods and insects and the cell walls of fungi.
  • Chitosan is water soluble, non-cytotoxic, readily absorbed through the intestine and mainly excreted in the urine.
  • the chitosan used herein has a low molecular weight. In one aspect, the chitosan has a molecular weight of less than 5,000 Da, less than 4,000 Da, less than 3,000 Da, less than 2,000 Da, or less than 1,500 Da. In another aspect, the chitosan has a molecular weight of 100 Da to less than 5,000 Da.
  • the chitosan has a molecular weight of 200 Da, 300 Da, 400 Da, 500 Da, 600 Da, 700 Da, 800 Da, 900 Da, 1,000 Da, 1,100 Da, 1,200 Da, 1,300 Da, 1,400 Da, 1,500 Da, 1,600 Da, 1,700 Da, 1,800 Da, 1,900 Da, or 2,000 Da, where any value can be a lower or upper endpoint of a range (e.g., 500 Da to 1,500 Da).
  • the chitosan can be deacetylated to a specific degree in order to enhance the hemostatic properties of the fluid complex coacervate.
  • the chitosan is deacetylated, ionizable primary amine groups are produced.
  • the amount of deacetylation it is possible to modify the positive charge density as needed of the chitosan.
  • the chitosan has a degree of acetylation greater than or equal to 80%, greater than or equal to 85%, greater than or equal to 90%, greater than or equal to 95%, or up to 100%.
  • the chitosan has a degree of deacetylation greater than or equal to 80% and has a molecular weight of less or equal to 2,000 Da. In another aspect, the chitosan has a degree of deacetylation greater than or equal to 90% and has a molecular weight of less or equal to 1,500 Da.
  • the polyanion can be a synthetic polymer or naturally-occurring.
  • naturally-occurring poly anions include glycosaminoglycans such as chondroitin sulfate, heparin, heparin sulfate, dermatan sulfate, keratin sulfate, and hyaluronic acid.
  • acidic proteins having a net negative charge at neutral pH or proteins with a low pi can be used as naturally- occurring polyanions described herein.
  • the anionic groups can be pendant to the polymer backbone and/or incorporated in the polymer backbone.
  • the polyanion is a synthetic polymer, it is generally any polymer possessing anionic groups or groups that can be readily converted to anionic groups by adjusting the pH.
  • groups that can be converted to anionic groups include, but are not limited to, carboxylate, sulfonate, boronate, sulfate, borate, phosphonate, or phosphate. Any cationic counterions can be used in association with the anionic polymers if the considerations discussed above are met.
  • the poly anionic component of the hemostatic complex coacervates described herein is polyphosphate, polyphosphonate, or a combination thereof.
  • a phosphate group has the general formula PO4 3 , where one or two oxygen atoms are ionizable (i.e., negatively charged oxygen atoms).
  • the phosphate group can be attached to a polymer via an phosphoester bond (i.e., -C-O- PO3 3 ).
  • the phosphate groups can be bonded to one another via phosphoester bonds (i.e., P-O-P).
  • a phosphonate group has the general formula C-PO 3 2 , where one or two oxygen atoms are ionizable (i.e., negatively charged oxygen atoms).
  • the phosphonate group via an organophosphorous bond.
  • the carbon atom in the formula C-PO 3 2 can be part of the polymer backbone or a sidechain bonded to the polymer backbone.
  • the polyphosphate and the polyphosphonate has from 5 to 100 mol % phosphate or phosphonate groups relative to the number of units or monomers comprising the polyphosphate or polyphosphonate. For example, if the
  • polyphosphate is a polyacrylate, and the polyacrylate is produced from 2 moles of acrylic acid and 3 moles of an acrylic monomer bearing a phosphate group, the resulting polyphosphate is composed of 60 mole % phosphate groups.
  • the polyphosphate can be a naturally-occurring compound such as, for example, DNA, RNA, or highly phosphorylated proteins like phosvitin (an egg protein), dentin (a natural tooth phosphoprotein), casein (a phosphorylated milk protein), or bone proteins (e.g. osteopontin), or phosphopeptides derived from these proteins.
  • phosvitin an egg protein
  • dentin a natural tooth phosphoprotein
  • casein a phosphorylated milk protein
  • bone proteins e.g. osteopontin
  • the polyphosphate can be a synthetic polypeptide made by polymerizing the amino acid serine and then chemically phosphorylating the polypeptide to produce a polyphosphoserine.
  • the polyphosphate can be a synthetic polypeptide made by polymerizing the amino acid serine and then chemically phosphorylating the polypeptide to produce a polyphosphoserine.
  • polyphosphoserine can be produced by the polymerization of the N-carboxy anhydride of phosphoserine.
  • the polyphosphate can be produced by chemically or enzymatically phosphorylating a protein (e.g., natural serine- or threonine-rich proteins).
  • the polyphosphate can be produced by chemically phosphorylating a polyalcohol including, but not limited to, polysaccharides such as cellulose or dextran.
  • the polyphosphate or polyphosphonate can be a synthetic compound.
  • the polyphosphate can be a polymer with pendant phosphate or phosphonate groups attached directly to the polymer backbone (i.e., no spacer between the polymer backbone and the phosphate group or phosphonate group) attached to the polymer backbone via a spacer.
  • the phosphate group(s) can be present in the polymer backbone (e.g., a phosphodiester backbone).
  • the polyphosphate or polyphosphonate includes a polyacrylate having one or more pendant phosphate or phosphonate groups.
  • the polyphosphate or polyphosphonate can be derived from the
  • the polyphosphate or polyphosphonate is a block co-polymer, where segments of the co-polymer possess phosphate or phosphonate groups or neutral groups depending upon the selection of the monomers used to produce the co polymer.
  • polyphosphate or polyphosphonate is a polymer having at least one fragment having the formula XI
  • R 4 is hydrogen or an alkyl group; n is from 1 to 10;
  • Y is oxygen, sulfur, or NR 30 , wherein R 30 is hydrogen, an alkyl group, or an aryl group;
  • Z’ is a phosphate group or a phosphonate group
  • Z’ in formula XI is phosphate and n in formulae XI is 2.
  • the polyphosphate is an inorganic polyphosphate possessing a plurality of phosphate groups (e.g., ⁇ ⁇ aP() ; ⁇ :; where n is 3 to 50).
  • n is 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, or 50, where any value can be a lower or upper endpoint of a range (e.g., 3 to 10, 10 to 20, etc.)
  • inorganic phosphates include, but are not limited to, Graham salts, hexametaphosphate salts, and triphosphate salts.
  • the counterion of these salts can be monovalent cations such as, for example, Na + , K + , and NH 4 + .
  • the polyphosphate is a phosphorylated sugar.
  • the sugar can be a hexose or pentose sugar. Additionally, the sugar can be partially or fully phosphorylated. In one aspect, the phosphorylated sugar is inositol hexaphosphate.
  • the preparation of the hemostatic complex coacervates described herein can be performed using a number of techniques and procedures.
  • An exemplary technique for producing a hemostatic complex coacervate is provided in the Examples.
  • the chitosan and polyphosphate are mixed as dilute solutions. Upon mixing, when the chitosan and polyphosphate associate they condense into a liquid phase at the bottom of a mixing chamber (e.g., a tube) to produce a condensed phase.
  • the condensed phase i.e., complex coacervate
  • an aqueous solution of poly cation is mixed with an aqueous solution of polyanion such that the positive/negative charge ratio of the poly cation to the polyanion is from 4 to 0.25, 3 to 0.25, 2 to 0.25, 1.5 to 0.5, 1.10 to 0.95, 1 to 1.
  • the amount of chitosan at a certain degree of deacetylation and polyanion can be varied in order to achieve specific
  • the hemostatic complex coacervate contains water, wherein the amount of water is from 20% to 80% by weight of the composition.
  • the pH of the solution containing the poly cation and the polyanion can vary in order to optimize complex coacervate formation.
  • the pH of the hemostatic complex coacervate is from 6 to 9, 6.5 to 8.5, 7 to 8, or 7 to 7.5.
  • the hemostatic complex coacervates described herein can be modified so that they form a gel in situ when administered to a subject.
  • These complex coacervates are referred to herein as in situ solidifying hemostatic complex coacervates.
  • the in situ solidifying hemostatic complex coacervate is a liquid at ionic strengths higher than the ionic strength at the site of administration, but insoluble ionic hydrogels at the ionic strength at the site of administration.
  • a solid or gel is formed in situ at the administration site as the salt concentration in the herein in situ solidifying hemostatic complex coacervates equilibrates to the administration site concentration.
  • the solid or gel that is subsequently produced is a non-fluid, water insoluble material.
  • the concentration of the monovalent ions in the in situ solidifying hemostatic complex coacervates is 0.5 M to 1.8 M, 0.5 M to 1.6 M, 0.5 M to 1.4 M, or 0.5 M to 1.2 M. In another aspect, the concentration of the monovalent ions in the in situ solidifying hemostatic complex coacervates is 1.5 to 2, 1.5 to 3, 1.5 to 4, 1.5 to 5, 1.5 to 6, 1.5 to 7, 1.5 to 8, 1.5 to 9 or 1.5 to 10 times greater than the concentration of the monovalent salt at the administration site.
  • a salt that produces monovalent ions when added to the poly cation and polyanion in water can be used to produce the in situ solidifying hemostatic complex coacervates described herein.
  • the monovalent salt used to prepare the in situ solidifying hemostatic complex coacervates is sodium chloride, sodium bromide, sodium fluoride, sodium pyruvate, sodium acetate, sodium tosylate, sodium benzenesulfonate, sodium benzoate, sodium lactate, sodium salicylate, sodium glucuronate, sodium galacturonate, sodium nitrite, sodium mesylate, sodium trifluoroacetate, sodium nitrate, sodium gluconate, sodium glycolate, sodium formate, or any combination thereof.
  • the hemostatic complex coacervates can optionally contain one or more multivalent cations (i.e.. cations having a charge of +2 or greater).
  • the multivalent cation can be a divalent cation composed of one or more alkaline earth metals.
  • the divalent cation can be a mixture of Ca +2 and Mg +2 .
  • transition metal ions with a charge of +2 or greater can be used as the multivalent cation.
  • the concentration of the multivalent cations can determine the rate and extent of coacervate formation. Not wishing to be bound by theory, weak cohesive forces between particles in the fluid may be mediated by multivalent cations bridging excess negative surface charges.
  • the amount of multivalent cation used herein can vary. In one aspect, the amount is based upon the number of anionic groups and cationic groups present in the poly cation and polyanion, respectively.
  • hemostatic complex coacervates can be formulated in hypertonic saline solutions that can be used for parenteral or intravenous
  • hemostatic complex coacervates can be formulated in Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, or other buffered saline solutions that can be safely administered to a subject.
  • the hemostatic complex coacervates described herein have numerous benefits and applications where it is desirable to reduce or inhibit bleeding in a subject.
  • the hemostatic complex coacervates are liquids with low viscosity and are readily injectable (i.e., flowable).
  • the hemostatic complex coacervates have other unique properties.
  • the hemostatic complex coacervates do not mix with blood.
  • the hemostatic complex coacervates can be applied to a pool of blood or areas where there are large volumes of blood without the concern of the hemostatic complex coacervate solubilizing in the blood.
  • the hemostatic complex coacervates are also denser than blood and can displace blood.
  • the hemostatic complex coacervates will displace the blood on the tissue and settle on the site of bleeding on the tissue.
  • the hemostatic complex coacervates are water-borne eliminating the need for potentially toxic solvents.
  • the hemostatic complex coacervates are biocompatible, non toxic, and biodegradable. Further in this aspect, the hemostatic complex coacervates can degrade and be eliminated from the body following its period of usefulness, but is stable enough against hydrolysis to last until hemostasis has begun or has been completed. In one aspect, the hemostatic complex coacervates do not cause allergic or sensitivity reactions. In another aspect, the hemostatic complex coacervates, when used, remain at or near body temperature (i.e., do not generate a significant amount of heat during any step of the hemostasis process).
  • the hemostatic complex coacervates described herein can inhibit or reduce bleeding internally in a subject or externally.
  • the source of bleeding is from a wound.
  • the wound can be caused by an accident due to an injury.
  • the source of bleeding can be the result of a surgical procedure.
  • the hemostatic complex coacervates described herein can inhibit or reduce bleeding from a number of different sites in a subject including, but not limited to, lacerations, punctures, scrapes, abrasions, or bleeding from a suture.
  • the hemostatic complex coacervates can be administered to the subject without forming a gel in situ.
  • an in situ solidifying hemostatic complex coacervate can be used to produce a gel upon administration.
  • the flowable hemostatic complex coacervates can be applied laproscopically, endoscopically, or can be physically administered to internal body cavities and organs prior to closing surgical incisions. Further in these aspects, hemostatic complex coacervates will not swell excessively and will not cause pain or discomfort for the patient.
  • the hemostatic complex coacervates have a low viscosity such that they can be introduced to the body in a minimally invasive way such as, for example, through a narrow gauge device, syringe, catheter, needle, cannula, or tubing.
  • the hemostatic complex coacervates can be applied directly at the site of internal bleeding such as, for example, at a biopsy site or an internal surgical incision.
  • the hemostatic complex coacervates can be applied internally to the gastrointestinal tract using an endoscope.
  • a gel can form at physiological ionic strength and/or when coming into contact with physiological fluids.
  • the hemostatic complex coacervates can be applied topically or externally to open wounds (e.g., applied by syringe or spraying), or can be applied to the gums and/or tongue following mouth injuries or oral surgeries to stop bleeding in the oral cavity.
  • the hemostatic complex coacervate can be applied to suture openings, which can further prevent bleeding.
  • the hemostatic complex coacervates are stable at room temperature and can be deployed instantly by emergency medical technicians and/or military medics to those injured on the battlefield, in accidents, and the like. In a further aspect, the hemostatic complex coacervates are sufficient to stop bleeding such that no further treatment is required. In an alternative aspect, the hemostatic complex coacervates can slow or reduce bleeding enough to stabilize a patient for transport to a hospital or field hospital for lifesaving treatment. In a still further aspect, the hemostatic complex coacervates can enhance the clotting process in individuals genetically lacking one or more factors necessary for proper hemostasis to occur (e.g., hemophiliac patients lacking von Willebrand factor or another clotting factor).
  • any of the hemostatic complex coacervates described herein can be applied to a solid substrate to produce a hemostatic substrate.
  • the substrate is a bandage, a sponge (e.g., GELFOAM ® ), a suture, a tissue graft, an implant, a stent, or a tape.
  • the hemostatic complex coacervate can be applied to the solid substrate using techniques in the art such as, for example, spraying or brushing the hemostatic complex coacervate on the solid substrate or dipping the solid substrate into the hemostatic complex coacervate.
  • a hemostatic composition comprises the hemostatic complex coacervate as described herein admixed with a granulated gelatin powder.
  • the granular gelatin powder is a porcine gelatin absorbable powder.
  • the powder can be GELFOAM ® manufactured by Pfizer or SURGIFOAM ® manufactured by Ethicon.
  • the hemostatic complex coacervate is from 1 vol% to 50 vol% of the hemostatic composition, or it is 1 vol%, 5 vol%, 10 vol%, 15 vol%, 20 vol%, 25 vol%, 30 vol%, 35 vol%, 40 vol%, 45 vol%, or 50 vol%, where any value can be a lower and upper end-point of a range (e.g., 5 vol% to 40 vol%, 10 vol%, 50 vol%, etc.).
  • the granular gelatin powder can be admixed with hemostatic complex coacervate using techniques known in the art to produce a paste that can be applied to the site of bleeding. Water or saline can optionally be added with the hemostatic complex coacervate and granular gelatin powder.
  • compositions, and methods described and claimed herein are made and evaluated, and are intended to be purely exemplary and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.) but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in °C or is at ambient temperature, and pressure is at or near atmospheric. There are numerous variations and combinations of reaction conditions, e.g., component concentrations, desired solvents, solvent mixtures, temperatures, pressures and other reaction ranges and conditions that can be used to optimize the product purity and yield obtained from the described process. Only reasonable and routine experimentation will be required to optimize such process conditions.
  • the following protocol describes the process of sodium hexametaphosphate (P6)/chitosan hemostatic complex coacervate made at a 1.2: 1 (-/+) charge ratio in NaCl concentration of 500 mM using 5M NaCl stock. Amounts are provided for a 10 mL; however, amounts can be adjusted accordingly for different batch sizes.
  • the pH of the stock solution is ⁇ 6.0 - 6.1, and the pH is not adjusted.
  • the hemostatic complex coacervate produced above was applied to a bleeding wound in a rat liver. Upon administration of the hemostatic complex coacervate via syringe to the wound, bleeding from the wound stopped shortly after administration.

Landscapes

  • Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Surgery (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Inorganic Chemistry (AREA)
  • Materials Engineering (AREA)
  • Engineering & Computer Science (AREA)
  • Composite Materials (AREA)
  • Materials For Medical Uses (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne des coacervats complexes hémostatiques fluides qui peuvent réduire ou inhiber le saignement chez un sujet. Les coacervats complexes sont composés d'un poly-cation et d'un poly-anion. Les coacervats complexes ont des propriétés qui sont supérieures par rapport aux agents hémostatiques classiques.
PCT/US2019/035923 2018-06-07 2019-06-07 Coacervats complexes hémostatiques fluides WO2019236932A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201862681853P 2018-06-07 2018-06-07
US62/681,853 2018-06-07

Publications (1)

Publication Number Publication Date
WO2019236932A1 true WO2019236932A1 (fr) 2019-12-12

Family

ID=68770786

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2019/035923 WO2019236932A1 (fr) 2018-06-07 2019-06-07 Coacervats complexes hémostatiques fluides

Country Status (1)

Country Link
WO (1) WO2019236932A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114522270A (zh) * 2021-12-22 2022-05-24 季华实验室 止血粉的制备方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016011028A1 (fr) * 2014-07-14 2016-01-21 University Of Utah Research Foundation Coacervats complexes de solidification in situ et leurs procédés de fabrication et d'utilisation
CN106474530A (zh) * 2015-08-24 2017-03-08 中国科学院金属研究所 一种基于壳寡糖的聚电解质海绵止血敷料的制备方法
WO2017152039A1 (fr) * 2016-03-04 2017-09-08 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Protection et administration de protéines thérapeutiques multiples

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016011028A1 (fr) * 2014-07-14 2016-01-21 University Of Utah Research Foundation Coacervats complexes de solidification in situ et leurs procédés de fabrication et d'utilisation
CN106474530A (zh) * 2015-08-24 2017-03-08 中国科学院金属研究所 一种基于壳寡糖的聚电解质海绵止血敷料的制备方法
WO2017152039A1 (fr) * 2016-03-04 2017-09-08 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Protection et administration de protéines thérapeutiques multiples

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114522270A (zh) * 2021-12-22 2022-05-24 季华实验室 止血粉的制备方法
CN114522270B (zh) * 2021-12-22 2023-02-21 季华实验室 止血粉的制备方法

Similar Documents

Publication Publication Date Title
US11471557B2 (en) In situ solidifying solutions and methods of making and using thereof
US11672756B2 (en) Temperature sensitive hydrogel composition including nucleic acid and chitosan
KR101121480B1 (ko) 미세다공성 입자를 포함하는 의학적으로 유용한 겔의 형성 및 사용 방법
JP2020527987A (ja) ヒアルロン酸−カルシウム及びポリリシンを含む創傷被覆材及びその製造方法
US20210162094A1 (en) Two-liquid type hemostatic composition and method for manufacturing the same
US12091471B2 (en) Methods and compositions for achieving hemostasis and stable blood clot formation
WO2013126550A2 (fr) Nouvelles compositions, leur préparation et leur utilisation
CN103189078A (zh) 含有透明质酸和l-精氨酸的抗粘连剂
US10077324B2 (en) Polymers, preparation and use thereof
WO2019236932A1 (fr) Coacervats complexes hémostatiques fluides
WO2019232823A1 (fr) Préparation pour microcapsule mélangée de chitosane et d'alginate de propylène glycol et son application
WO2019147922A2 (fr) Appareil et procédé d'utilisation de coacervats complexes à solidification in situ pour une occlusion vasculaire
JP3825411B2 (ja) リンゴ酸ハイドロゲル
AU2019212513B2 (en) Apparatus and method of using in situ solidifying complex coacervates for vascular occlusion
CN118546437A (zh) 一种含羧甲壳聚糖和氧化透明质酸的水凝胶及其制备方法
KR20240095440A (ko) 합성 외과용 지혈제
WO2013166616A1 (fr) Biomatériau comprenant du chitosane et au moins un glycosaminoglycane

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19816030

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205N DATED 24.03.2021)

122 Ep: pct application non-entry in european phase

Ref document number: 19816030

Country of ref document: EP

Kind code of ref document: A1