WO2024042535A1

WO2024042535A1 - Hemostatic composite, its method and applications thereof

Info

Publication number: WO2024042535A1
Application number: PCT/IN2023/050784
Authority: WO
Inventors: Jayanta Haldar; Rajib DEY
Original assignee: Jawaharial Nehru Centre for Advanced Scientific Research
Current assignee: Jawaharial Nehru Centre for Advanced Scientific Research
Priority date: 2022-08-20
Filing date: 2023-08-18
Publication date: 2024-02-29
Anticipated expiration: 2025-02-20

Abstract

The present disclosure relates to a hemostatic porous composite comprising: a. a water- absorbing polymer matrix having a plurality of anti-microbial agent with a quaternary 5 ammonium group; and b. a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the water-absorbing polymer matrix is impregnated with and is bound to the plurality of inorganic material. The present disclosure further provides a process for the preparation of the hemostatic porous composite. Furthermore, the present disclosure provides a kit, a pharmaceutical 10 composition and an article comprising the hemostatic porous composite as disclosed herein.

Description

HEMOSTATIC COMPOSITE, ITS METHOD AND APPLICATIONS THEREOF

TECHNICAL FIELD

[0001] The present disclosure relates to the field of biomaterials, specifically, to hemostatic materials and their application in sealing active bleeding and preventing wound infections. In particular, the present disclosure relates to antimicrobial hemostatic sponges (BACSTAT), that exhibit superior mechanical property, hemostatic ability and potent antimicrobial activity. The present disclosure further relates to a method of its preparation and applications thereof.

BACKGROUND OF INVENTION

[0002] Hemorrhage control is a noteworthy concern for martial and civilian trauma centers throughout the world (Johnson, D. et al. Ann. Med Surg. 2014, 3, 21). The unrestrained bleeding after accident or surgery results in more than 5.8 million deaths worldwide annually (Rossaint, R. et al. Crit Care. 2016, 20, 100). Hence, employing an efficient hemostatic material is of crucial importance during the trauma emergency to hold the bleeding rapidly and effectively. Concurrently, drug-resistant bacterial and fungal infection associated with this combat trauma wound, further deteriorates the situation, resulting in delayed wound healing (Mao, X. et al. Adv. Sci. 2019, 6, 1801555). While many hemostatic agents are existing in the market, they are found to be unable to achieve the clinical necessity, especially for the noncompressible wounds or the deep, irregularly shaped wounds experienced in both battlefield and post-surgery in clinical settings (Zhao, X. et al. Nat. Commun. 2018, 9, 2784 and Zheng, W. et al. Chem. Eng. J. 2022447, 137482). Notably, they are ineffective to tackle the infection as well.

[0003] Therefore, an ideal dressing material with anti-infection, hemostatic, and wound healing properties is the need of the hour to treat, specifically, trauma-associated wounds.

SUMMARY OF THE INVENTION

[0004] In an aspect of the present disclosure, there is provided a hemostatic porous composite comprising: (i) a water-absorbing polymer matrix having a plurality of antimicrobial agent with a quaternary ammonium group; and (ii) a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material.

[0005] In another aspect of the present disclosure, there is provided a process for preparation of a hemostatic porous composite as disclosed herein, comprising: a waterabsorbing polymer matrix having a plurality of anti-microbial agent with a quaternary ammonium group; and a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material, the process comprising: a. mixing the functionalized compound, the monomer of the compound of Formula I, and the antimicrobial agent of a compound of Formula II with the plurality of inorganic material to obtain a reaction mixture; b. curing the reaction mixture to obtain a crosslinked polymerized product; and c. lowering a temperature followed by lyophilizing the crosslinked polymerized product to obtain a hemostatic porous composite.

[0006] In yet another aspect of the present disclosure, there is provided a method for treating an injury in a subject, the method comprising: administering the hemostatic porous composite comprising: a water-absorbing polymer matrix having a plurality of anti-microbial agent with a quaternary ammonium group; and a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material, to an injury site; and allowing the hemostatic porous composite to swell and seal at the injury site.

[0007] In one another aspect of the present disclosure, there is provided a kit, comprising: the hemostatic porous composite comprising: a water-absorbing polymer matrix having a plurality of anti-microbial agent with a quaternary ammonium group; and a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material; and instructions for using the hemostatic porous composite.

[0008] In a further aspect of the present disclosure, there is provided a pharmaceutical composition comprising the hemostatic porous composite comprising: a water-absorbing polymer matrix having a plurality of anti-microbial agent with a quaternary ammonium group; and a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material, together with a pharmaceutically acceptable carrier, optionally in combination with other therapeutic agents.

[0009] In more aspect of the present disclosure, there is provided an article comprising hemostatic porous composite comprising: a water-absorbing polymer matrix having a plurality of anti-microbial agent with a quaternary ammonium group; and a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material.

[0010] These and other features, aspects, and advantages of the present subject matter will be better understood with reference to the following description. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter.

BRIEF DESCRIPTION OF THE ACCOMPANYING FIGURES

[0011] In order that the disclosure may be readily understood and put into practical effect, reference will now be made to exemplary embodiments as illustrated with reference to the accompanying figures. The figures together with a detailed description below, are incorporated in and form part of the specification, and serve to further illustrate the embodiments and explain various principles and advantages, in accordance with the present disclosure wherein:

[0012] Figure 1 depicts a general procedure of BACSTAT sponge preparation, in accordance with an implementation of the present disclosure.

[0013] Figure 2 depicts characterization of BACSTAT sponges, A) IR (Infra-red) spectra of BACSTAT-8 by comparing with i) Dextran -Methacrylate (Dex-MA) and ii) mesoporous silica Santa Barbara Amorphous (SBA-15); B) (i) scanning electron microscopic (SEM) images of the sponges, (ii) SEM images showing the morphology of sponges at different states such as in normal state, compressed state, and self-healing phases after swelling, (iii) pore size of BACSTAT-8 sponge and (iv) porosity% of BACSTAT-8 sponge; C) digital images showing the swelling property of compressed BACSTAT-8 sponge in water; i) by immersing in excess water; ii) by immersing in 2 ml water, iii) swelling ratio of the BACSTAT-8 sponges and iv) fluid absorpotion ratio of the BACSTAT-8 sponges, in accordance with an implementation of the present disclosure.

[0014] Figure 3 depicts the i) digital images of cyclic compression test of BACSTAT-8 sponge in TA rheometer, ii) compression property of sponges, iii) cyclic compression test of BACSTAT-8 sponges, in accordance with an implementation of the present disclosure.

[0015] Figure 4 depicts the bactericidal activity of BACSTAT sponges by counting method with the number of viable bacteria after treatment with sponges, in accordance with an implementation of the present disclosure.

[0016] Figure 5 depicts the antibacterial activity of BACSTAT sponges in various bacterial strains i) by dragging method and ii) by visual turbidity method, and iii) antibacterial activity of BACSTAT- 1 sponge, in accordance with an implementation of the present disclosure.

[0017] Figure 6 depicts the bactericidal kinetics of sponges against i) MRS A R3545; ii) A. baumannnii MTCC1425; iii) P. aeruginosa MTCC424, in accordance with an implementation of the present disclosure.

[0018] Figure 7 depicts the antibacterial activity of sponges against stationary phase bacterial cell, in accordance with an implementation of the present disclosure.

[0019] Figure 8 depicts the fungicidal activity of sponges against clinically isolated C. albicans strains, in accordance with an implementation of the present disclosure.

[0020] Figure 9 depicts hemolysis of sponges, i) digital images of blood cells soup after treatment with sponges; ii) percentage of hemolysis of the sponges, in accordance with an implementation of the present disclosure.

[0021] Figure 10 depicts in vivo biocompatibility of sponges (control and BACSTAT-8) by incorporation into subcutaneous pocket for 72 h, in accordance with an implementation of the present disclosure.

[0022] Figure 11 depicts hemostatic ability of BACSTAT sponges, A) In vitro hemostatic ability of BACSTAT-8; i) digital images of blood clotting experiment with and without water; ii) Blood Clotting indices (BCI) of sponges; iii) SEM images of blood cells upon treatment with BACSTAT-8; and B) In vivo hemostatic ability of BACSTAT- 8; i) Live images of blood clotting of BACSTAT-8 sponge in mice liver puncture model; Blood loss in ii) mice liver puncture model; and iii) mice femoral vein injury model, in accordance with an implementation of the present disclosure.

[0023] Figure 12 depicts percentage of A) red blood cells adhesion; B) platelets adhesion, in accordance with an implementation of the present disclosure.

DETAILED DESCRIPTION OF THE INVENTION

[0024] Those skilled in the art will be aware that the present disclosure is subject to variations and modifications other than those specifically described. It is to be understood that the present disclosure includes all such variations and modifications. The disclosure also includes all such steps, features, compositions, and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any or more of such steps or features.

Definitions

[0025] For convenience, before further description of the present disclosure, certain terms employed in the specification, and examples are delineated here. These definitions should be read in the light of the remainder of the disclosure and understood as by a person of skill in the art. The terms used herein have the meanings recognized and known to those of skill in the art, however, for convenience and completeness, particular terms and their meanings are set forth below.

[0026] The articles “a”, “an” and “the” are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.

[0027] The terms “comprise” and “comprising” are used in the inclusive, open sense, meaning that additional elements may be included. It is not intended to be construed as “consists of only”.

[0028] Throughout this specification, unless the context requires otherwise the word “comprise”, and variations such as “comprises” and “comprising”, will be understood to imply the inclusion of a stated element or step or group of element or steps but not the exclusion of any other element or step or group of element or steps.

[0029] The term “including” is used to mean “including but not limited to”. “Including” and “including but not limited to” are used interchangeably. [0030] In the structural formulae given herein and throughout the present disclosure, the following terms have been indicated meaning, unless specifically stated otherwise.

[0031] The term “at least one” is used to mean one or more and thus includes individual components as well as mixtures/combinations.

[0032] The term “hemostatic sponge”, “hemostatic porous composite”, “BACSTAT sponge”, and “hemostatic materials” are used interchangeably.

[0033] The term “hemostatic porous composite” as used herein, denotes porous materials that carry superior mechanical properties which when applied to the wound site can quickly swell to seal and apply compression to the deep and non-compressible wounds. These materials also exhibit the anti-infection property against the microbes, including drug-resistant microbes, such as bacteria, fungi, virus, etc.

[0034] The term “plurality of inorganic material” as used herein, denotes one or more of same or different types of inorganic materials can be employed during the process of crosslinking of a functionalized compound, a monomer of a compound of Formula I, and an anti-microbial agent of a compound of Formula II, as disclosed herein. The plurality of inorganic material for the purpose of the present disclosure is selected from a group consisting of SBA-15, MCM-41, laponite, and combinations thereof.

[0035] The term “functionalized compound” as used herein, denotes that the monomers employed in the polymer matrix of the present disclosure are modified with polar and nonpolar functionalities, in order to achieve the objectives, in terms of superior mechanical property. For the purpose of the present disclosure, the functionalized compound comprises acrylate functionalized dextran, methacrylate functionalized dextran, acrylate functionalized cellulose, methacrylate functionalized cellulose, acrylate functionalized alginate, methacrylate functionalized alginate, acrylate functionalized chitosan, methacrylate functionalized chitosan, acrylate functionalized gelatin, methacrylate functionalized gelatin, or combinations thereof.

[0036] The phrase “the polymer matrix is impregnated with and is bound to the plurality of inorganic material” as used herein, denotes that the plurality of inorganic material provide a surface for the polymer matrix to undergo cross-linking and such polymer matrix is impregnated with and is bound covalently or/and non-covalently to the plurality of inorganic material. [0037] The term “degree of substitution” as used herein, denotes the average level of substituent groups attached per base unit.

[0038] The terms “injury site” and “wound site” are used interchangeably. Said terms as disclosed herein denote physiological damage caused to any part of the body that results in unrestrained bleeding. Such damage may be caused due to an accident or a result of surgery.

[0039] The term “pharmaceutically acceptable carrier” refers to the carriers or vehicle compounds used in the process of drug delivery which serves to improve the selectivity, effectiveness, and/or safety of drug administration majorly by controlled release of drugs into the systemic circulation of the administered subject.

[0040] The term “anti-inflammatory agents” as used herein, denotes the drugs or reagents that reduce inflammation in the body. The anti-inflammatory agents of the present disclosure include but not limited to aspirin, ibuprofen, nimesulide, naproxen, acetaminophen, dexamethasone, or combinations thereof.

[0041] The term “wound healing promoting agents” as used herein, denotes the drugs or reagents that replace the damaged tissues with newly formed tissues and thereby resulting in healing the wound. The wound healing promoting agents of the present disclosure include but not limited to proline, mucin, collagen, vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), fibrinogen growth factor (FGF), or combinations thereof.

[0042] The term “antioxidants” as used herein, denotes the drugs or reagents that have the capability to interact with and neutralize free radicals in the body and thereby prevent them from damage. The antioxidants of the present disclosure include but not limited to glutathione, ascorbic acid, or combinations thereof.

[0043] The term “antiseptics” or “antiseptic agents” as used herein, denotes the drugs or reagents that are applied on the wound site to prevent infections. The antiseptics of the present disclosure include but not limited to chlorhexidine, chloroxylenol, povidone- iodine, or combinations thereof.

[0044] The term “antiparasitics” or “antiparasitic agents” as used herein, denotes the drugs or reagents that are used for the treatment and management of infections caused by a wide class of parasites. The antiparasitics of the present disclosure include but not limited to chloroquine, doxycycline, nitazoxanide, benznidazole, paromomycin sulfate, pyrimethamine, or combinations thereof.

[0045] The term “antifungal agents” as used herein, denotes the drugs or reagents that are used for the treatment and prevention of infections caused by fungi. The antifungal agents of the present disclosure include but not limited to beclometasone, clotrimazole, fluconazole, amphotericin, ketoconazole, nystatin, terbinafine, clotrimazole, or combinations thereof.

[0046] The term “antibiotic agents” as used herein, denotes the drugs or reagents that are active against a wide spectrum of bacteris, including gram-positive and gram-negative bacteria. The antibiotic agents of the present disclosure include but not limited to vancomycin, cephalosporins, neomycin, chloramphenicol, fusidic acid, colistin, or combinations thereof.

[0047] A term once described, the same meaning applies for it, throughout the disclosure. [0048] 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 disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosure, the preferred methods, and materials are now described. All publications mentioned herein are incorporated herein by reference.

[0049] As discussed in the background, there is a need for a hemostatic material that provides superior hemostasis and can also be fabricated into a variety of shapes that are suitable for use in sealing and controlling bleeding from a variety of wounds. At the same time, the hemostatic material shall also impose anti-infection properties to prevent the wound site against wide range of drug-resistant microbes including bacteria, fungi, protozoa, virus, etc.

[0050] To address the aforementioned issues, various hemostatic technologies have been developed. Recently, hemostatic device has been developed which is an injector system packed with several compressed cellulose sponges which can rapidly expand to seal and apply pressure to the deep, non-compressible wounds. Similarly, various other shape memory cryogels, sponges, polymer foams have been developed as hemostatic agents or wound dressings materials. However, the known hemostatic materials fail to address the problems associated with the immediate chance of infection caused by drug-resistant microbes, such as bacteria, fungi, virus at the accidental injury site or wound site. Hence, an ideal dressing material with shape memory mechanical property, hemostatic ability and most importantly anti-infection property is of current need. Moreover, it is indeed necessary to have an ideal dressing material (sponge) with an economically viable synthetic procedure that can additionally tackle a broad spectrum of pathogens.

[0051] Towards this goal, the present disclosure relates to the development of antimicrobial hemostatic sponge with superior mechanical properties which can swiftly swell to seal and apply compression to deep and non-compressible wounds, once applied. Said hemostatic sponge shall also impose the anti-infection property against the drugresistant pathogens. Thus, the present disclosure provides a hemostatic porous composite comprising a polymer matrix which includes a plurality of anti-microbial agent with a quaternary ammonium group and the polymer matrix is impregnated with and is bound to the plurality of inorganic material. Further, the polymer matrix is a cross-linked polymerized product of: a functionalized compound; a monomer of a compound of Formula I

Formula I wherein,

Ri is selected from hydrogen, Ci-6 alkyl, or C2-6 alkenyl;

R2 is selected from hydrogen, -CO-C1-6 alkyl, or -CO-C2-6 alkenyl; n' is an integer selected from 1 to 500; and an anti-microbial agent of a compound of Formula II:

Formula II wherein,

R’ and R” are independently selected from hydrogen or Ci-6 alkyl;

R3 is selected from hydrogen, C1-6 alkyl, or C2-6 alkenyl;

X is selected from -O-, -N-, or -S-;

R4 is selected from hydrogen, Ci-30 alkyl, C2-30 alkenyl, or O ; wherein Y is selected from -NH or -O-;

Rs is selected from Ci-30 alkyl or C2-30 alkenyl; m is an integer selected from 2 to 6; in a presence of the plurality of inorganic material. The hemostatic porous composite of the present disclosure can rapidly seize the bleeding in the trauma site or would site by activating the platelet and closing the wound fast by forming the new blood vessel.

[0052] In an embodiment of the present disclosure, there is provided a hemostatic porous composite comprising: i. a water-absorbing polymer matrix having a plurality of antimicrobial agent with a quaternary ammonium group; and ii. a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material. In another embodiment of the present disclosure, the plurality of inorganic material is in a weight range of 10-25% with respect to the porous composite. In another embodiment of the present disclosure, the plurality of inorganic material is in a weight range of 15- 20% with respect to the porous composite. In still another embodiment of the present disclosure, the plurality of inorganic material is in a weight range of 17 to 18%, and preferably 17.2% with respect to the porous composite. [0053] In an embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the anti-microbial agent with a quaternary ammonium group is a compound of Formula II:

Formula II wherein,

R’ and R” are independently selected from hydrogen or Ci-6 alkyl;

R3 is selected from hydrogen, C1-6 alkyl, or C2-6 alkenyl;

X is selected from -O-, -N-, or -S-;

R4 is selected from hydrogen, C1-30 alkyl, C2-30 alkenyl, or O ; wherein Y is selected from -NH- or -O-;

Rs is selected from Ci-30 alkyl or C2-30 alkenyl; and m is an integer selected from 2 to 6.

[0054] In another embodiment of the present disclosure the anti-microbial agent with a quaternary ammonium group is a compound of Formula II:

Formula II wherein,

R’ and R” are independently hydrogen or C1-3 alkyl;

R3 is selected from hydrogen, C1-3 alkyl, or C2-6 alkenyl; X is -0-;

R4 is selected from hydrogen, Ci-10 alkyl, C2-10 alkenyl, or O ; wherein Y is selected from -NH- or -O-;

Rs is selected from Ci-10 alkyl or C2-10 alkenyl; and m is an integer selected from 2 to 4.

[0055] In another embodiment of the present disclosure the anti-microbial agent with a quaternary ammonium group is a compound of Formula II:

Formula II wherein,

R’ and R” are independently Ci alkyl;

R3 is Ci alkyl;

X is -O-;

R4 is selected from hydrogen, Ci, alkyl, Cs alkyl, or C10 alkyl, m is an integer selected from 2 to 3.

[0056] In further embodiment of the present disclosure the anti-microbial agent with a quaternary ammonium group is a compound of Formula II:

Formula II wherein, R’ and R” are independently CH3;

R3 is CH₃;

X is -0-;

R4 is Ce alkyl, Cs alkyl, or C10 alkyl, m is 2.

[0057] In an embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the polymer matrix is a cross-linked polymerized product of: a. a functionalized compound; b. a monomer of a compound of Formula I:

Formula I wherein,

Ri is selected from hydrogen, C1-6 alkyl, or C2-6 alkenyl;

R2 is selected from hydrogen, -CO-C1-6 alkyl, or -CO-C2-6 alkenyl; n' is an integer selected from 1 to 500; and c. an anti-microbial agent of a compound of Formula II:

Formula II wherein,

R’ and R” are independently selected from hydrogen or C1-6 alkyl;

R₃ is selected from hydrogen, C1-6 alkyl, or C2-6 alkenyl;

X is selected from -O-, -N-, or -S-; R4 is selected from hydrogen, Ci-30 alkyl, C2-30 alkenyl, or

wherein Y is selected from -NH- or -O-;

Rs is selected from Ci-30 alkyl or C2-30 alkenyl; m is an integer selected from 2 to 6, in a presence of the plurality of inorganic material.

[0058] In an embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the polymer matrix is a cross-linked polymerized product of: a. a functionalized compound; b. a monomer of a compound of Formula I:

Formula I wherein,

Ri is selected from hydrogen, or C1-6 alkyl;

R2 is selected from hydrogen, or -CO-C2-6 alkenyl; n' is an integer selected from 1 to 500; and c. an anti-microbial agent of a compound of Formula II:

Formula II wherein, R’ and R” are independently Ci-6 alkyl;

R3 is Ci-10 alkyl;

X is -0-;

R4 is selected from hydrogen, Ci-10 alkyl, C2-10 alkenyl or

wherein Y is selected from -NH- or -O-;

Rs is selected from Ci-10 alkyl or C2-10 alkenyl; and m is an integer selected from 2 to 3, in presence of the plurality of inorganic material.

[0059] In an embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the polymer matrix is a cross-linked polymerized product of: a. a functionalized compound; b. a monomer of a compound of Formula I:

Formula I wherein,

Ri is hydrogen, C1-6 alkyl, or C2-6 alkenyl;

R2 is -CO-C2-6 alkenyl; n' is an integer selected from 1 to 500; and c. an anti-microbial agent of a compound of Formula II:

Formula II wherein,

R’ and R” are independently Ci-io alkyl;

R₃ is CH₃;

X is -0-;

R4 is C 1-10 alkyl, m is 2, in presence of the plurality of inorganic material.

[0060] In an embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the polymer matrix is a cross-linked polymerized product of: a. a functionalized compound; b. a monomer of a compound of Formula I:

Formula I wherein,

Ri is hydrogen;

R2 is -CO-C2 alkenyl; n' is an integer selected from 1 to 500; and c. an anti-microbial agent of a compound of Formula II:

Formula II wherein,

R’ and R” are independently CH₃; R₃ is CH₃;

X is -0-;

R4 is Ci alkyl, Ce alkyl, Cs alkyl, or C10 alkyl, m is 2, in presence of the plurality of inorganic material.

[0061] In an embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the functionalized compound comprises acrylate functionalized dextran, methacrylate functionalized dextran, acrylate functionalized cellulose, methacrylate functionalized cellulose, acrylate functionalized alginate, methacrylate functionalized alginate, acrylate functionalized chitosan, methacrylate functionalized chitosan, acrylate functionalized gelatin, methacrylate functionalized gelatin, or combinations thereof. In another embodiment of the present disclosure the functionalized compound comprises acrylate functionalized dextran, methacrylate functionalized dextran, or combinations thereof.

[0062] In an embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the functionalized compound is of Formula A:

Formula A, wherein,

R is independently selected from hydrogen, or -CO-C2-6 alkenyl; and n is an integer selected from 50 to 15000.

[0063] In another embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the functionalized compound is of Formula A:

Formula A, wherein,

R is independently selected from hydrogen, or -CO-C2-4 alkenyl; and n is an integer selected from 50 to 15000.

[0064] In still another embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the functionalized compound is of Formula A:

Formula A, wherein,

R is independently selected from hydrogen, or -CO-C2 alkenyl; and n is an integer selected from 50 to 15000

[0065] In an embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the functionalized compound is of Formula A:

Formula A, wherein,

R is independently selected from hydrogen, or -CO-C2-6 alkenyl; and n is an integer selected from 50 to 15000, wherein the functionalized compound of Formula A has a degree of substitution in a range of 1% to 50%. In yet another embodiment, the functionalized compound of Formula A has a degree of substitution in a range of 5% to 25%. In another embodiment, the functionalized compound of Formula A has a degree of substitution in a range of 6% to 8%.

[0066] In an embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the plurality of inorganic material is selected from a group consisting of SBA-15, MCM-41, laponite, and combinations thereof. The plurality of inorganic material is SBA-15.

[0067] In another embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the polymer matrix is cross-linked with the functionalized compound in a weight percentage range of 1-60%, the monomer of the compound of Formula I in a weight percentage range of 1-60%, and the anti-microbial agent with a quaternary ammonium group of Formula II in a weight percentage range of 1-60% in the presence of the plurality of inorganic material in a weight percentage range of 1-50%, with respect to the total weight of the porous composite. In another embodiment of the present disclosure, the polymer matrix is cross-linked with the functionalized compound in a weight percentage range of 5-50%, the monomer of the compound of Formula I in a weight percentage range of 5-50%, and the anti-microbial agent with a quaternary ammonium group of Formula II in a weight percentage range of 5-50%, in the presence of the plurality of inorganic material in a weight percentage range of 5-40%, with respect to the total weight of the porous composite. In yet another embodiment of the present disclosure, the polymer matrix is cross-linked with the functionalized compound in a weight percentage range of 15-30%, the monomer of the compound of Formula I in a weight percentage range of 15-30%, and the anti-microbial agent with a quaternary ammonium group of Formula II in a weight percentage range of 15-30%, in the presence of the plurality of inorganic material in a weight percentage range of 10-30%, with respect to the total weight of the porous composite. In still another embodiment of the present disclosure, the polymer matrix is cross-linked with the functionalized compound in a weight percentage range of 27-28%, the monomer of the compound of Formula I in a weight percentage range of 27-28%, and the anti-microbial agent with a quaternary ammonium group of Formula II in a weight percentage range of 27-28%, in the presence of the plurality of inorganic material in a weight percentage range of 16-18%, with respect to the total weight of the porous composite.

[0068] In another embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the polymer matrix is cross-linked with the functionalized compound in a weight percentage range of 1-60%, the monomer of the compound of Formula I in a weight percentage range of 1-60%, the anti-microbial agent with a quaternary ammonium group of Formula II in a weight percentage range of 1-60% in the presence of the plurality of inorganic material in a weight percentage range of 1 - 50% with respect to the total weight of the porous composite, and the polymer matrix is cross-linked by photocuring, or thermal curing.

[0069] In an embodiment of the present disclosure, there is provided a a hemostatic porous composite as disclosed herein, the composite comprising a functionalized compound (dextran methacrylate, Dex-MA), a monomer of a compound of Formula I (polyethelene glycol diacrylate, PEG-DA), a antimicrobial agent of a compound of Fomula II (lipophilic methacrylate-octane, LipMA-CsHn) and a plurality of inorganic material (mesoporous silica, SBA-15) in a weight ratio of 4:4:4:2.5.

[0070] In another embodiment of the present disclosure, there is provided a hemostatic porous composite as disclosed herein, wherein the composite has a pore size in a range of 1 pm to 100 pm and a specific surface area in a range of 1 m²/kg to 50 m²/kg. In yet another embodiment, the composite has a pore size in a range of 5 pm to 50 pm and specific surface area in a range of 5 m²/kg to 40 m²/kg. In still another embodiment of the present disclosure, the composite has a pore size in a range of 8 pm to 13 pm and specific surface area in a range of 5 m²/kg to 40 m²/kg.

[0071] In an embodiment of the present disclosure, there is provided a hemostatic porous composite comprising: i. a water-absorbing polymer matrix having a plurality of antimicrobial agent with a quaternary ammonium group; and ii. a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material; and the composite possesses both hemostasis and antimicrobial property. [0072] In an embodiment of the present disclosure, there is provided a hemostatic porous composite comprising: i. a water-absorbing polymer matrix having a plurality of antimicrobial agent with a quaternary ammonium group; and ii. a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material, and the composite optionally comprises at least one therapeutic agent selected from the group consisting of antimicrobial agents, anti-inflammatory agents, wound healing promoting agents, anti-oxidant agents, antibiotic agents, antiseptics, antiparasitics, antifungal agents and combinations thereof.

[0073] In an embodiment of the present disclosure, there is provided a process for preparation of hemostatic porous composite as disclosed herein, the process comprising: a. mixing the functionalized compound, the monomer of the compound of Formula I, and the anti-microbial agent of a compound of Formula II with the plurality of inorganic material to obtain a reaction mixture; b. curing the reaction mixture to obtain a crosslinked polymerized product; and c. lowering a temperature followed by lyophilizing the crosslinked polymerized product to obtain a hemostatic porous composite.

[0074] In an embodiment of the present disclosure, there is provided a process of preparation of hemostatic porous composite as disclosed herein, wherein the reaction mixture is stirred for 30 min to 24 h at a temperature in a range of 4 °C to 50 °C in presence of a solvent. The solvent is selected from water or buffer. Furthermore, the buffer is selected from 0.9 % saline, phosphate buffer saline (PBS, pH=7-7.2), 2-(N- morpholino) ethanesulfonic acid (MES) buffer (pH= 7-7.2), Bis-tris buffer (pH= 7-7.2), or Citrate buffer (pH=6.2). Preferably, the solvent is water. In another embodiment of the present disclosure, the reaction mixture is stirred for 5 h to 22 h at a temperature in a range of 20°C to 40 °C in presence of a solvent. In yet another embodiment, the reaction mixture is stirred for about 10 h to 20 h at a temperature in a range of 25°C to 35 °C in presence of a solvent. In still another embodiment, the reaction mixture is stirred for about 12 h to 14 h at room temperature in presence of water as solvent.

[0075] In an embodiment of the present disclosure, there is provided a process of preparation of hemostatic porous composite as disclosed herein, wherein the lowering the temperature is carried out in a presence of liquid nitrogen for a time period of 5 to 60 mins. In a further embodiment, the lowering the temperature is carried out in a presence of liquid nitrogen for a time period of 5 to 30 mins.

[0076] In an embodiment of the present disclosure, there is provided a process of preparation of hemostatic porous composite as disclosed herein, wherein the curing is carried out by photocuring in presence of a photo -initiator selected from 2-Methyl-l-[4- (methylthio)phenyl]-2-(4-morpholinyl)-l -propanone (irgacure 907), or 2-Hydroxy-l-[4- (2-hydroxyethoxy)-phenyl]-2-methyl-l -propanone (irgacure 2959); or by thermal curing using ammonium persulphate (APS) or N,N,N',N'-Tetramethylethylenediamine (TEMED). Preferably, the curing is photocuring in presence of a photo-initiator selected from irgacure 907 or irgacure 2959.

[0077] In an embodiment of the present disclosure, there is provided a process for preparation of hemostatic porous composite as disclosed herein, the process comprising: a. mixing the functionalized compound, the monomer of the compound of Formula I, and the anti-microbial agent of a compound of Formula II with the plurality of inorganic material to obtain a reaction mixture; b. curing the reaction mixture to obtain a crosslinked polymerized product; and c. lowering a temperature followed by lyophilizing the crosslinked polymerized product to obtain a hemostatic porous composite, wherein the functionalized compound is in a concentration range of 1 mg/mL to 100 mg/mL, the monomer of the compound of Formula I is in a concentration range of 1 mg/mE to 100 mg/mL, and the anti-microbial agent of a compound of Formula II is in a concentration range of 1 mg/mL to 100 mg/mL, and the plurality of inorganic material is in a concentration range of 1 mg/mL to 100 mg/mL, in the finally obtained hemostatic porous composite.

[0078] In an embodiment of the present disclosure, there is provided a method for treating an injury in a subject, the method comprising: administering the hemostatic porous composite as disclosed herein to an injury site; and allowing the hemostatic porous composite to swell and seal at the injury site.

[0079] In an embodiment of the present disclosure, there is provided a method for treating an injury in a subject as disclosed herein, wherein the composite imparts hemostasis and antimicrobial property at the injury site. [0080] In an embodiment of the present disclosure, there is provided a method for treating an injury in a subject as disclosed herein, wherein the composite swell when it comes in contact with a fluid and has a swelling ratio in a range of 1.5 to 50 times compared to the initial size. The fluid is selected from water, blood, urine, or stimulated body fluid. In another embodiment of the present disclosure, the composite exhibits a swelling ratio % in a range of 300 to 600% in a fluid. In still another embodiment of the present disclosure, the composite exhibits a swelling ratio % in a range of 350 to 500% in water and a swelling ratio% in a range of 300 to 400% in blood.

[0081] In an embodiment of the present disclosure, there is provided a method for treating an injury in a subject as disclosed herein, wherein the injury is selected from a wound, a hemorrhage, a hematoma, or a bleeding tissue.

[0082] In an embodiment of the present disclosure, there is provided a kit as disclosed herein, the kit comprising: the hemostatic porous composite comprising: i. a waterabsorbing polymer matrix having a plurality of anti-microbial agent with a quaternary ammonium group; and ii. a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material; and instructions for using the hemostatic porous composite.

[0083] In an embodiment of the present disclosure, there is provided a pharmaceutical composition as disclosed herein, the pharmaceutical composition comprising: the hemostatic porous composite comprising: i. a water-absorbing polymer matrix having a plurality of anti-microbial agent with a quaternary ammonium group; and ii. a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material, together with a pharmaceutically acceptable carrier, optionally in combination with other therapeutic agents. The therapeutic agents are selected from antimicrobial agents, anti-inflammatory agents, wound healing promoting agents, antioxidant agents, antibiotic agents, antiseptics, antiparasitics, antifungal agents or combinations thereof.

[0084] In an embodiment of the present disclosure, there is provided an article comprising the hemostatic porous composite comprising: i. a water-absorbing polymer matrix having a plurality of anti-microbial agent with a quaternary ammonium group; and ii. a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material. The article is selected from compressed porous tablets, dressing mat, or injectable compressed sponge. Preferably, the hemostatic porous composite of the present disclosure is in the form of injectable compressed sponge.

[0085] In an embodiment of the present disclosure, there is provided a use of the hemostatic porous composite as disclosed herein; the kit as disclosed herein; the pharmaceutical composition as disclosed herein; or the article as disclosed herein, for the manufacture of medical aid compositions.

[0086] In an embodiment of the present disclosure, there is provided a use of the hemostatic porous composite as disclosed herein; the kit as disclosed herein; the pharmaceutical composition as disclosed herein; or the article as disclosed herein, for the treatment of injuries in a subject.

[0087] In an embodiment of the present disclosure, there is provided a use of the hemostatic porous composite as disclosed herein; the kit as disclosed herein; the pharmaceutical composition as disclosed herein; or the article as disclosed herein, for the treatment of injuries in a subject, wherein the injury is selected from a wound, a hemorrhage, a hematoma, or a bleeding tissue.

[0088] In an embodiment of the present disclosure, there is provided a use of the hemostatic porous composite comprising: i) a water-absorbing polymer matrix having a plurality of anti-microbial agent with a quaternary ammonium group; and ii) a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the polymer matrix is impregnated with and is bound to the plurality of inorganic material; the kit comprising the hemostatic porous composite as disclosed herein and instructions for using the same; the pharmaceutical composition comprising the hemostatic porous composite as disclosed herein together with a pharmaceutically acceptable carrier, optionally in combination with other therapeutic agent; or the article comprising hemostatic porous composite as disclosed herein, for the treatment of injuries in a subject. [0089] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations are possible.

EXAMPLES

[0090] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply.

[0091] Materials and methods: Reagent grade chloroform (CHCI3), methanol (MeOH), dimethyl sulphoxide (DMSO) were obtained from Spectrochem (India). The solvents were dried prior to use wherever necessary. Dextran, glycidyl methacrylate, poly(ethelene glycol) diacrylate, 2-(dimethylamino)ethyl methacrylate, poly(ethylene glycol)-block-poly(propylene glycol) -block-poly (ethylene glycol) (pluronic P123), tetraethyl orthosilicate, 1 -bromohexane, 1 -bromooctane, 1 -bromodecane were procured from Sigma- Aldrich. Potassium carbonate and cone. H2SO4 were purchased from SD Fine Chemical Ltd (India). These chemicals were directly used for reaction. Nuclear magnetic resonance (NMR) spectra were recorded in deuterated solvents employing Bruker AMX-400 spectrometer. Rheological and mechanical properties were recorded in TA rheometer (DHR-3) instrument. FT-IR spectra were recorded on a Bruker IFS 66v/S spectrometer used in attenuated total reflectance Fourier transform (ATR FT-IR) mode. Customized UV-curing chamber was used for coating. Tecan Infinite M200 PRO Microplate Reader was used for measurement of optical density (O.D.). Bacterial strains, Pseudomonas aeruginosa MTCC424, and Acinetobacter baumannii MTCC1425 were purchased from MTCC (Chandigarh, India). Methicillin -resistant Staphylococcus aureus (MRSA) ATCC33591, and Enterococcus faecium ATCC19634 were obtained from ATCC (Rockville, MD, USA). MRSA R3889, P. aeruginosa R590, and A. baumannii R674 were obtained from National Institute of Mental Health and Neurosciences, Bangalore, India. Vancomycin -resistant S. aureus 4 (VRSA 4) and vancomycin-resistant E. faecium 903 (VRE 903) were obtained from Central Drug Research Institute (CDRI). C. albicans AB226 and C. albicans AB399 were acquired from Anthem Biosciences, Bangalore, India. RAW 264.7 (ATCC TIB- 71) cell lines were procured from ATCC (Rockville, MD, USA). Growth media and agar for bacteria, fungus, and mammalian cells were obtained from HIMEDIA, India. The Lactate dehydrogenase (LDH) kit including the cell lyse solution was brought from Thermo Fisher scientific.

Example 1

Example 1.1. Synthesis of Dextran Methacrylate (Dex-MA) (functionalized compound):

[0092] Dextran (from Leuconostoc spp., Mr ~ 40 kDa) (5 g, 30.8 mmol) was dissolved in anhydrous dimethyl sulphoxide (DMSO) (100 mL) at room temperature under the nitrogen atmosphere. After dissolving the polymer, 4-dimethylaminopyridine (1 g, 8.2 mmol) was added to the solution. After 2 h of stirring, glycidyl methacrylate (876 mg, 6.1 mmol) was added to it. The reaction was allowed to stir for 48 h at room temperature under dark condition. Finally, the reaction solution was subjected to extensive dialysis for about 3 days to replace DMSO and other low molecular weight impurities. The dialysate was then lyophilized and the obtained solid product was stored at -20 °C for further use. The degree of substitution was determined by ¹ H-NMR analysis from the ratio of integration of methacrylate proton and C-l ring proton.

[0093] Dex-MA: ^XH-NMR (400 MHz, CDC1₃): 8 6.25 (CH₂=C(CH₃)CO-), 5.82 (CH₂=C(CH₃)CO-), 5.01 (Cl-H, 1H), 3.521-4.023 (C2-, C3-, C4-, C5-, and C6-H, 6H), 1.99 (CH₂=CH(CH₃)CO-, 3H);

[0094] FT-IR (v): 3098-3565 cm’¹ (OH str.), 2907 cm’¹ (CH₂ assym. str.), 1706 cm’¹ (C=O str.), 1631 cm’¹ (C=C str.), 845 cm’¹ (C=C-H ben.) Example 1.2. Synthesis of lipophilic methacrylate (LipMA) monomer (an antimicrobial agent of a compound of Formula II):

[0095] In a solution of (N,N-dimethyl)aminoethylmethacrylate (15 mmol) in dry chloroform (15 mL), various alkyl bromides, e.g., methyl bromide, hexyl bromide, octyl bromide, and decyl bromide (22.5 mmol) were added individually and the reaction solutionswere allowed to stir at 65 °C for 48 h. Subsequently, the reaction solutions were allowed to attain room temperature and transferred to a round bottom flask. The total volume of the reaction mixtures was then further reduced to l/10th of the original volume by using the rotary evaporator. The products were precipitated using excess of diethyl ether (150 mL). The excess organic solvent was then decanted off and the precipitates were washed repeatedly (three times) using diethyl ether followed by evaporating the solvent using a high vacuum pump to get the final products with 90-95% yield. All the compounds were characterized by ’ H-NMR and high-resolution mass spectroscopy (HRMS).

[0096] LipMA-CeH : ‘ H-NMR (400 MHz, CDC1₃): 6 6.128 (CH₂=C(CH₃)CO-, d, 1H), 5.660 (CH₂=C(CH₃)CO-,d, 1H), 4.562-4.611 (CH₂-O-CO-, t, 2H), 4.147-4.153 (-O-CH₂-CH₂-N⁺(CH₃)₂-, t, 2H), 3.612-3.689 (-CH₂-N⁺(CH₃)₂-CH₂, t, 2H), 3.524 (- N⁺(CH₃)₂-CH₂, S, 6H), 1.931 CH₂=C(CH₃)CO-, S, 3H), 1.728-1.742 (-N⁺(CH₃)₂-CH₂- CH₂-, m, 2H), 1.225-1.312 (-N⁺(CH₃)₂-CH₂- (CH₂)₃CH₃, m, 6H), 0.841-0.876 (- N⁺(CH₃)₂-CH₂-(CH₂)₅CH₃, t, 3H). HRMS: calculated m/z 242.2115 [M-Br’]⁺; observed m/z 242.2122 [M-Br’]⁺.

[0097] LipMA-CsHn: ’ H-NMR (400 MHz, CDC1₃): 6 6.134 (CH₂=C(CH₃)CO-, d, 1H), 5.663-5.67 (CH₂=C(CH₃)CO-,d, 1H), 4.637-4.647 (CH₂-O-CO-, t, 2H), 4.162- 4.172 (-O-CH₂-CH₂-N⁺(CH₃)₂-, t, 2H), 3.634 (-CH₂-N⁺(CH₃)₂-CH₂, t, 2H), 3.515 (- N⁺(CH₃)₂-CH₂, S, 6H), 1.940 CH₂=C(CH₃)CO-, S, 3H), 1.749-1.767 (-N⁺(CH₃)₂-CH₂- CH₂-, m, 2H), 1.242-1.333 (-N⁺(CH₃)₂-CH₂- (CH₂)₅CH₃, m, 10H), 0.846-0.888 (- N⁺(CH₃)₂-CH₂-(CH₂)₅CH₃, t, 3H). HRMS: calculated m/z 270.2428 [M-Br’]⁺; observed m/z 270.2437 [M-Br’]⁺.

[0098] LipMA-CioH₂i: ’ H-NMR (400 MHz, CDC1₃): 6 6.148 (CH₂=C(CH₃)CO-, d, 1H), 5.689 (CH₂=C(CH₃)CO-, d, 1H), 4.6421-4.6647 (CH₂-O-CO-, t, 2H), 4.1689- 4.1923 (-O-CH₂-CH₂-N⁺(CH₃)₂-, t, 2H), 3.581-3.6236 (-CH₂-N⁺(CH₃)₂-CH₂, t, 2H), 3.509 (-N⁺(CH₃)₂-CH₂, S, 6H), 1.957 CH₂=C(CH₃)CO- s, 3H), 1.628 (-N⁺(CH₃)₂- CH₂-CH₂- m, 2H), 1.255-1.349 (-N⁺(CH₃)₂-CH₂-(CH₂)₇CH₃, m, 14H), 0.869-0.895 (-N⁺(CH₃)₂-CH₂-(CH₂)₅CH₃, t, 3H). HRMS: calculated m/z 298.2741 [M-Br’]⁺; observed m/z 298.2761 [M-Br“]⁺.

Example 1.3. Synthesis of mesoporous silica (SBA-15) (inorganic material):

[0099] Synthesis of Santa Barbara Amorphous (SBA-15) was followed from a well- established protocol. Briefly, 4 g of pluronic P123 surfactant was dissolved in 105 ml of deionized water and 10.8 ml of cone. H₂SO4 followed by stirring at 40 °C for 3 h. During this step, the surfactant pluronic P123 formed spherical micelles which act as soft templates for directing the structure. Subsequently, 9.063 ml of tetraethyl orthosilicate (TEOS) was added to this solution and allowed to stir for 3.5 h with a higher stirring rate to avoid gelling. During this step, the spherical micelles elongate to give a cylindrical hexagonal array of micelles. The uniform solution after stirring in the round bottom flask was fitted with a condenser and kept for hydrothermal treatment at 100 °C for 30 h without stirring. During the hydrothermal treatment, the micelles expand to mesoporous range. After the treatment, the solution was filtered and washed repeatedly with deionized water to eliminate the extra surfactant. The obtained white powder was then dehydrated at 100 °C overnight and finally, it was calcined for 6 h at 550 °C to remove the organic template.

Example 2. Synthesis of hemostatic sponge (BACSTAT) (hemostatic porous composite):

[00100] In this present disclosure, four different kinds of BACSTAT sponges were prepared. The flow chart illustrating the general scheme for the preparation the hemostatic porous composite of the present disclosure is given in Figure 1. According to the general procedure, Dextran Methacrylate (Dex-MA, 160 mg) was dissolved in 1 mL Millipore water. 160 mg of cationic lipophilic methacrylate monomer (LipMA) was independently dissolved in 1 mL millipore water. The aqueous solution of another monomer PEG-DA was prepared at 160 mg/mL concentration. On the other side, 100 mg SBA-15 was suspended in 1 mL milipore water. All the four aqueous compositions were mixed to make a total volume of 4 mL and was allowed to stir vigorously for overnight. The so obtained well-mixed reaction mixture (4 mL) was then transferred in a 10 mL glass test tube (borosil) and mixed with 4 mg of photo -initiator Iragcure 2959 (final cone. = 1 mg/mL). The resulting solution was then UV crosslinked with 365 nm (5 mW cm^-2) for 10 min to obtain a crosslinked polymerized product. The crosslinked polymerized product hydrogel was immediately freezed down in liquid N2 for 10 minutes and finally lyophilized for 48 h to remove the water crystal, resulting in the respective hemostatic porous composite or hemostatic sponge.

[00101] BACSTAT-0 was produced by following the aforementioned protocol through crosslinking of Dex-MA, PEG-DA and SBA-15 (Dex-MA : PEG-DA : SBA-15 = 4:4:2.5 wt %).

[00102] BACSTAT-1 was produced by following the aforementioned protocol through crosslinking of Dex-MA, PEG-DA, LipMA-CH₃ and SBA-15 (Dex-MA : PEG- DA : LipMA-CH₃ : SBA-15 = 4:4:4:2.5 wt %).

[00103] BACSTAT-6 was produced by following the aforementioned protocol through crosslinking of Dex-MA, PEG-DA, LipMA-C₆Hi₃ and SBA-15 (Dex-MA : PEG-DA : LipMA-C₆Hi₃ : SBA-15 = 4:4:4:2.5 wt %).

[00104] BACSTAT-8 was produced by following the aforementioned protocol through crosslinking of Dex-MA, PEG-DA, LipMA-CsHn and SBA-15 (Dex-MA : PEG-DA : LipMA-CsHn : SBA-15 = 4:4:4:2.5 wt %).

[00105] BACSTAT-10 was produced by following the aforementioned protocol through crosslinking of Dex-MA, PEG-DA, LipMA-CioH_2i and SBA-15 (Dex-MA : PEG-DA : LipMA-CioH_2i : SBA-15 = 4:4:4:2.5 wt%).

[00106] Additionally, other three compositions of BACSTAT-8 sponge were prepared for further enhancement of swelling ratio and fluid absorption properties. According to the general procedure, various composition of Dex-MA, LipMA-CsHn, PEG-DA, and SBA-15 were dissolved in 4 mL Millipore water followed by stirring vigorously for overnight. Then the well- mixed reaction mixture (4 mL) was transferred into a 10 mL glass test tube (borosil) and mixed with 4 mg of photo -initiator Iragcure 2959 (final cone. = 1 mg/mL). The resulting solution was then UV crosslinked with 365 nm radiation (5 mW cm^-2) for 10 min. The crosslinked polymerized product hydrogel so obtained was immediately freezed down in liq N2 for 10 minutes and finally lyophilized for 48 h to remove the water crystal, resulting in the respective hemostatic porous composite or sponge.

[00107] BACSTAT-8-a was produced by following the aforementioned protocol through crosslinking of Dex-MA, PEG-DA, LipMA-CsHn and SBA-15 (Dex-MA : PEG-DA : LipMA-CsHn : SBA-15 = 4:4:4:4 wt %).

[00108] BACSTAT-8-b was produced by following the aforementioned protocol through crosslinking of Dex-MA, PEG-DA, LipMA-C8H17 and SBA-15 (Dex-MA : PEG-DA : LipMA-CsHn : SBA-15 = 4:2:2: 1 wt %).

[00109] BACSTAT-8-c was produced by following the aforementioned protocol through crosslinking of Dex-MA, PEG-DA, LipMA-C8H17 and SBA-15 (Dex-MA : PEG-DA : LipMA-CsHn : SBA-15 = 4:2:2:4 wt%).

[00110] The hemostatic porous composites obtained by the process as disclosed in Examples 1 to 2, were then employed to obtain a kit with instructions for using the hemostatic porous composites.

[00111] Further, the hemostatic porous composites obtained by the process as disclosed herein were used to prepare a pharmaceutical composition along with an appropriate pharmaceutical carrier and a therapeutic agent seleted from antimicrobial agents, antiinflammatory agents, wound healing promoting agents, anti-oxidant agents, antibiotic agents, antiseptics, antiparasitics, antifungal agents or combinations thereof. The pharmaceutical composition or the porous composite was used for the treatment of an injury, a wound, a hemorrhage, a hematoma, or a bleeding tissue. The porous composite or the pharmaceutical composition was applied directly on the injury and allowed to swell and seal at the injury site. The composite swell when it comes in contact with a fluid selected from water, blood, urine, or stimulated body fluid and has a swelling ratio in a range of 1.5 to 50 times compared to the initial size. The porous composite imparts hemostasis and antimicrobial property at the injury site.

Example 3. Characterization of hemostatic porous composite (hemostatic sponge) of the present disclosure: Example 3.1. FT-IR of the sponges:

[00112] The BACSTAT sponges were characterised using the infrared spectroscopy to analyse the presence of the functionalized compound Dex-MA and the inorganic material SB A- 15. Figure 2 A (i) and (ii) provides the infrared spectra of sponges recorded by attenuated total reflectance (ATR) sampling method on IFS6 V/s spectrometer (Bruker). From the Figure 2 A (i), the peaks corresponding to carbonyl stretching frequencies at 1729cm ¹ and 1706 cm'¹ were found as comparable to that of the Dex-MA. Further, Figure 2A (ii) shows that the prominent peaks of SBA-15 (the appearance of the Si-0 asymmetric stretching frequency at 1085 cm-1 and the Si-O-Si bending at 490 cm-1) were visible in the IR peaks of BACSTAT-8.

Example 3.2. Scanning Electron Microscopy of sponges:

[00113] The BACSTAT sponges were characterised using the scanning electron microscopy to analyse the topographical changes brought to the porous composite upon addition of antimicrobial agent of compound of Formula II LipMA-CsHn in comparison with the sponge without antimicrobial agent. The Figure 2 B (i) shows the cross- sections of sponges that were utilized for field emission scanning electron microscopy (FESEM) studies. Each sample was gold- sputtered before imaging. Further, the morphology of sponges at different states such as in normal state, compressed state, and self-healing phases after swelling were determined. From SEM images (Figure 2B (ii)) it was found that in compressed state, the sponge was less porous in structure compared to the normal phase, whereas when this compressed sponge expanded in water, it become porous again like initial state. To determine the pore size of the sponges, cross-sections of sponges were utilized for FESEM studies. Each sample was gold-sputtered before imaging. From SEM images the pore size was determined using ImageJ software. The pore size of BACSTAT-8 was found to be in the range of 8-13 pm (Figure 2 B (iii)).

[00114] To investigate the percentage of porosity of the sponges, 8 mm diameter sponges were immersed into excess volume of water. The absorption of water was measured and the porosity of sponges was investigated by following the equation below.

„ (Weight of water soaked sponge-Weight of initial sponge) Porosity (%)= - — - : - - - —; - , ■ ■ ■ , - * 100 %

Density of water * Volume of initial sponge [00115] From Figure 2B (iv), more than 65% of BACSTAT-8 structure were found porous in nature.

Example 3.3. Swelling property of sponges:

[00116] BACSTAT sponges with 8 mm diameter and 8 mm length were axially compressed. Then these compressed sponges were immersed into excess volume of water and human blood. The swelling of these compressed sponges was monitored and the digital images were captured in Figure 2C (i) and (ii). The increase in volume was calculated by following the equation-

(Volume of swelled sponge-Volume of compressed sponge)

Swelling ratio (%)= - — - - - - - x lOO %

Volume of compressed sponge

[00117] The fluid absorption by the sponge was calculated from the following equation- z (Weight of swelled sponge-Weight of compressed sponge)

Fluid absorption ratio (%)= - . , - - - _; / 100 %

Weight of compressed sponge

[00118] The swelling ratio and fluid absorption property of BACSTAT-8 sponges in different fluids are represented in Figure 2C (iii) and (iv) respectively. The B ACSTAT- 8 sponges exhibited appreciable fluid absorption ratio of 800 to 1100% in water and 500 to 700% in blood. In addition, the swelling ratio of the BACSTAT-8 sponges was found to be 350 to 500% in water and 300 to 400% in blood.

[00119] Furthermore, the swelling property and fluid absorption properties of the composites BACSTAT-8-a, BACSTAT-8-b, and BACSTAT-8-c were found to be lesser than those of the BACSTAT-8 sponge. Hence, BACSTAT-8 sponge was found to be the optimized hemostatic porous composite.

Example 3.4. Mechanical property of sponges:

[00120] The mechanical performance of the sponges were analysed by compression test and cyclic compression test using TA rheometer (DHR-3) at room temperature, as shown in Figure 3. The sponges were cut into cylindrical shapes with a diameter of 8 mm and height of 6 mm. The sponges were firstly swollen in DI (deionized) water, and then the compression test of the wet sponges were conducted with the maximal compression strain up to 80% at a strain speed was 100 pm/s. For the cyclic compression test, a drop of water was added around the wet sponge sample on the platform before the test, and then an 80% compression strain was employed to conduct the cyclic compression test. The compression strain was firstly performed up to 80% strain and then released to 0% strain by using constant compression and release strain rate, which was repeated for several times. In Figure 3, a total of 6 cycles data have been represented.

Example 4: Bactericidal activity of BACSTAT-8 sponge by counting method:

[00121] At first, sponges were cut into 8 mm diameter shape and transferred into 48 wells plate. Then, the 6 h grown mid-log phase bacterial culture was diluted to 10⁵ CFU/mL in I *PBS. Afterward, 100 pL of this bacterial suspension was inoculated and spread eventually onto the sponge. The sponges were then incubated for 3 h at 37 °C. Post incubation, 100 pL of IxPBS was used to wash by crushing sponges. Consequently, the suspensions were 10-fold serially diluted followed by drop-plated onto agar plates. Finally, the plates were incubated at 37 °C for 24 h to count the viable bacterial colony. Figure 4 depicts the bactericidal activity of BACSTAT sponges by counting method with the number of viable bacteria after treatment with sponges, wherein asterisk symbol indicates complete killing, the detection limit of the experiment was 50 CFU/mL, and VRSA stands for vancomycin-resistant S. aureus. BACSTAT-0 was found to be inactive to kill the bacterial burden. BACSTAT-6 displayed moderate bactericidal activity, whereas BACSTAT-8 and BACSTAT- 10 completely killed the challenged bacterial burden.

Example 5. Bactericidal activity of BACSTAT sponges against planktonic bacteria

[00122] At first, sponges were cut into 8 mm diameter shape initially and transferred into 48 wells plate. Then, the 6 h grown mid-log phase bacterial culture (planktonic phase) was diluted to 10⁵ CFU/mL in IxPBS (phosphate buffered saline). Afterward, 100 pL of this bacterial suspension was inoculated and spread eventually onto the sponge. The sponges were then incubated for 3 h at 37 °C. The antibacterial activity of the sponges was determined in two procedures- i) dragging method; and ii) visual turbidity method. In dragging method, (Figure 5 (i)), after incubation, sponges were dragged along the diameter and placed on nutrient agar plates. Then these plates were incubated at 37 °C for 18 hrs followed by imaging of the plates. In visual turbidity method (Figure 5(ii)), after incubation with bacteria for 3 h, sponges were directly immersed in fresh culture media and allowed to grow for 6 h at 37 °C followed by imaging of the visual turbidity. [00123] From Figure 5, the effective performance of the hemostatic porous composite of the present disclosure could be observed. It is clearly illustrated that BACSTAT-0 was inactive, as there was no crosslinked cationic moiety (LipMA, antimicrobial agent of a compound of Formula II). This could be understood from the bacterial growth in the dragged area (Figure 5(i)) and complete turbidity in Figure 5(ii), for BACSTAT-0. BACSTAT-1 was found to be inactive to kill the tested pathogens as shown in Figure 5(iii). BACSTAT-6 showed moderate bactericidal activity, whereas, BACSTAT-8 and BACSTAT-10 showed complete killing against all tested bacterial strains.

Example 6. Bactericidal kinetics of sponge against planktonic bacteria

[00124] Sponges were cut into 8 mm diameter shape initially and transferred into 48 wells plate. Then, the 6 h grown mid-log phase bacterial culture (planktonic phase) was diluted to 10⁵ CFU/mL in l ^zPBS. Afterward, 100 pL of this bacterial suspension was inoculated and spread eventually onto the sponge followed by incubation at different time intervals (0, 1, 2, and 3 h) at 37 °C. At different time intervals, bacteria incubated with sponges were serially diluted and drop-platted on nutrient agar plate to get the bacterial count.

[00125] The bactericidal kinetics of the hemostatic sponges of the present disclosure in Figure 6 clearly demonstrated that the BACSTAT-8 was able to kill completely the challenged bacterial burden of MRS A R3545, A. baumannii MTCC 1425, and P. aeruginosa MTCC424 within 2 h. Therefore, the composite of the present disclosure exhibit excellent antimicrobial activity against a wide range drug-resistant microbes, including bacteria.

Example 7. Bactericidal activity of BACSTAT sponges against stationary phase bacteria [00126] Sponges were cut into 8 mm diameter shape initially and transferred into 48 wells plate. Then, from 6 h grown culture, 3 pL suspension was diluted in 3 mL nutrient broth and incubated at 37 °C for 16 h. Then, stationary phase bacteria were diluted to 10⁵ CFU/mL in UPBS. Afterward, 100 pL of this bacterial suspension was inoculated and spread eventually onto the sponge. The sponges were then incubated for 3 h at 37 °C. The antibacterial activity of the sponges was determined by the dragging method. Figure 7 provides the dragging method, wherein, after incubation, sponges were dragged along the diameter and placed on nutrient agar plates. Then these plates were incubated at 37 °C for 18 hrs followed by imaging of the plates. It is clearly shown that BACSTAT-8 and BACSTAT-10 showed complete killing against all tested bacterial strains, whereas BACSTAT-0 was found inactive.

Example 8. Antifungal activity of BACSTAT sponges

[00127] At first, sponges were cut into 8 mm diameter shape. Besides, Candida albicans were spread-platted on yeast extract-peptonedextrose (YPD) agar plates from glycerol stock kept at -80 °C and incubated at 30 °C for 24 h. A single colony was then grown with 3 mL YPD medium at 30 °C for 10 h. After that, these mid-log phase grown fungi were diluted to 10⁵ CFU/mL in 1* PBS. Afterward, 100 pL of the fungi suspension was incubated with sponge for 3 h. After incubation, 100 pL of 1 * PBS was given on the sponge to wash by crashing it. Consequently, the antifungal activity was determined by counting the viable cells. The suspensions were ten-fold serially diluted and each dilution was drop-plated onto YPD agar. The plates were then incubated for 48 h at 37 °C and no of fungal colonies were counted.

[00128] Figure 8 proved the effective antifungal activity of the hemostatic composite of the present disclosure against Candida albicans. BACSTACT-6, BACSTAT-8 and BACSTAT-10 were shown to have antifungal activity and on the other hand, BACSTAT-0 was not active. Surprisingly, that BACSTAT-8 and BACSTAT-10 were shown to have unexpected and superior antifungal activity.

Example 9. Hemolytic activity of BACSTAT sponges against human erythrocytes. [00129] After cutting the sponges in 8 mm diameter, these were transferred into 48 well plates. By collecting the blood from a healthy donor, red blood cells (hRBCs) were isolated from blood by centrifugation at 3500 rpm for 5 min. The RBC was then resuspended in lx PBS to make the final concentration of 5 vol%. 300 pL of this suspension was then added by spreading on the sponges. 50 pL PBS was taken as a negative control and 0.1 vol% solution of Triton X-100 (TRX, 50 pL) was taken as the positive control. The plates were then incubated for 1 h at 37 °C. Next, cells were centrifuged and 100 pL of supernatants from the wells were relocated to a new 96-well plate for recording absorbance at 540 nm. Consequently, the percentage of hemolysis was calculated from the equation: (A-Ao)/(Atotai-Ao) X 100%, where A denotes the absorbance for the tested samples, A_o is the absorbance for the wells containing only water and erythrocytes suspension, and A_totai represents the absorbance of completely lysed cells (treated with Triton-X), all measured at 540 nm.

[00130] From Figure 9, it was observed that BACSTAT-6 and BACSTAT-8 sponges revealed 3% and 5-6 % of hemolysis respectively. Whereas, BACSTAT-10 showed 24% hemolysis.

Example 10. In-vivo toxicity of BACSTAT sponge

[00131] In-vivo toxicity of BACSTAT sponge was evaluated by incorporating the sponge inside the subcutaneous pocket for 72 h. In brief, male BALB/c mice (age 6-8 weeks) was used for the experiments. Before the experiment, the dorsal area of mice was shaved clearly. Then 8 diameter sponges were incorporated in the subcutaneous pocket. Without any sponge, vacant subcutaneous pocket was taken as control. After 72 h, the subcutaneous layer was cut properly and stored in 10% formalin solution to perform the histological analysis. Figure 10 represents the histopathological data, wherein the arrow indicates the epithelium layer; asterisks indicate the inflammatory cells infiltration, SG stands for subcutaneous gland, HF stands for hail follicles, and AT stands for adipose tissue. In Figure 10, BACSTAT-8 treated subcutaneous layer showed the normal architecture of epidermis containing stratified squamous epithelial cells with cytoplasm and nucleus, lined by keratin layer, the fibrous tissue with oval shaped nucleus, and also containing sweat and sebaceous glands, and hair follicles, adipose tissue containing fat cells with oval nucleus. These results were similar to the untreated normal skin and affirmed the biocompatibility of BACSTAT-8 sponge.

Example 11 In-vitro hemostatic ability of BACSTAT sponge

Example 11.1. Determination of blood clotting index (BCD:

[00132] Heparinised blood (HB) was prepared by collecting blood from healthy C57BL/6 mice in heparin (75 USP units) containing vacutainer. 50 pL of HB was gradually dropped upon the sponge placed in a petri-dish and incubated by shaking gently at 37 °C. After 5 min, 10 mL of millipore water was added carefully to the petri-dish without disturbing the clotted blood. Only HB without any material was taken as a reference. The blood clotting index (BCI) was evaluated by measuring the relative absorbance of 200 pL of blood sample which was diluted with 10 mL of water at 541 nm wavelength. The BCI of various materials was calculated using the following equation

ODmaterial ~ Absorbance of HB incubated with the material after dilution

ODreference ~ Absorbance of HB without any material after dilution

[00133] BACSTAT-8 revealed potent blood clotting efficiency which has been confirmed from Figure HA(i) and HA(ii).

Example 11.2. Visualization of the blood cells clotting on the sponge:

[00134] A small piece (8 mm diameter) of the sponge (BACSTAT-8) was incubated with 50 pL of the whole blood collected from a healthy donor for 5 min at 37 °C. After incubation, the sponge incubated with blood was washed with dPBS three times to remove the un-adhered cells. Then, 2.5% glutaraldehyde solution was used for another 2 h to fix the cells. The sample was air-dried and finally dried overnight in high vacuum. After that, the dried sample was further imaged using the FESEM technique after gold sputtering to visualize the blood cells onto the sponge after incubation. BACSTAT-8 has activated the platelets to clot the blood fast (Figure 11 A(iii)).

Example 11.3. Hemostatic ability in mice liver puncture model: [00135] To estimate the in-vivo hemostatic ability of sponge, the mice liver puncture model (BALB/c mice, 17-20 g, 6 weeks, male) was used. Mice were sedated by IP injection of the brew of ketamine and xylazine. The mice’s liver was open very carefully by abdominal incision, serous fluid was removed very gently and the liver was kept on a pre-weighed filter paper. The liver was then punctured using a 16 G needle and pre-weighted BACSTAT-8 sponge was applied to the incision site immediately. Mice, fixed on the corkboard, were tilled by 45° angle to increase the blood flow through the filter paper. After 2 min, the filter paper with absorbed blood was weighted and compared with the control group without any treatment. Sterile gauze was also used as a reference. Figure 11 B(i) and B(ii) clearly illustrate that BACSTAT-8 sponge seized the blood loss efficiently in mice liver incision model.

Example 11.4. Hemostatic ability in mice femoral vein incision model:

[00136] The BALB/c male mice (6-8 weeks, 18-22 g) were anesthetized by using a cocktail of ketamine-xylazine and fixed on a surgical corkboard. The ventral part of the mouse leg was incised to expose the femoral vein, and a 2 mm incision was made at the femoral vein. Blood was collected, and the amount was measured. The blood collection for the control case (in a centrifuge tube using the pipette) was done until the natural clotting happened (approximately 2-3 min). In the case of treatment, pre-weighted BACSTAT sponges were applied immediately after the injury was created and held there for 2 min. After that, the weight of the blood-soaked sponges was quantified to evaluate the blood loss in sponge-treated cases. Four mice were used in each group for this study. BACSTAT-8 sponge seized the blood loss efficiently in mice liver incision model in Figure 11 B(iii).

Example 12: Adhesion test

[00137] The adhesion of the blood cells upon the obtained sponges were evaluated.

Red blood cell adhesion test:

[00138] The adhesion of human red blood cells (hRBCs) onto the materials (sponge or gauge) was evaluated by measuring the absorbance of hemoglobin. Briefly, 50 pL of freshly collected human RBCs was incubated with the materials for 15 min at 37 °C. After incubation, the non-adherent hRBCs were washed out by rinsing the materials with IxPBS. Subsequently, the RBCs attached onto the material surface were lysed with 1 mL Milli-Q water by incubating for 1 h at 37 °C. The absorbance of all sample solutions was determined at 540 nm wavelength. 50 pL of lysed hRBCs was taken for the reference. The percentage of the attached hRBCs was measured by the following formula-

ODmaterial

[00139] Attachment of RBCs (%) = xl00%, wherein ODmateriai refers to the

ODreference optical density of the material and ODreference refers to the optical density of the reference. [00140] The adhesion of RBCs on BACSTAT-8 materials are represented in Figure 12A, which showed that the porous composite BACSTAT-8 exhibited an appreciable hRBC adhesion of about 60%.

Platelet adhesion test:

[00141] The attachment of platelet cells was evaluated by measuring the lactate dehydrogenase (LDH) activity of the adhered platelet. Firstly, the platelet rich plasma (PRP) was prepared by centrifuging the heparinized blood at 1500 rpm at 4 °C for 10 min. About 50 pL of thus obtained PRP was incubated with the materials (sponges and gauge) for 30-45 min at 37 °C. After that, materials were rinsed with IxPBS to wash out the non-attached platelets. Then, the Lactate dehydrogenase (LDH) kit including the cell lyse solution was used to determine the lactate dehydrogenase activity of the adhered platelets by following the manufacturers’ protocol. 50 pL of PRP without materials was taken as the reference control. The relative optical density (OD490- OD690) was measured by using a Tecan microplate reader. The percentage of the attached platelet was calculated by the subsequent formula-

[00142] Attachment of platelet (

[00143] The platelet adhesions on BACSTAT-8 sponge and gauze are represented in Figure 12B, which showed that the porous composite BACSTAT-8 exhibited a highly appreciable platelet adhesion of about 90%.

Advantages of the present disclosure [00144] The above-mentioned implementation examples as described on this subject matter and its equivalent thereof have many advantages, including those which are described.

[00145] The hemostatic porous composite, also called as hemostatic sponge of the present disclosure exhibit superior mechanical property, hemostatic ability and excellent antimicrobial activity against a wide range drug-resistant microbes including bacteria, fungi, and virus. The hemostatic porous composite of the present disclosure quickly swell to seal and apply compression to the deep and non-compressible wounds. It can rapidly seize the bleeding in the trauma site by activating the platelet and close the wound fast by forming a new blood vessel.

[00146] Although the subject matter has been described in considerable detail with reference to certain embodiments thereof, other embodiments are possible. As such, the spirit and scope of the disclosure should not be limited to the description of the embodiments contained herein.

Claims

I/We Claim:

1. A hemostatic porous composite comprising: i. a water-absorbing polymer matrix having a plurality of anti-microbial agent with a quaternary ammonium group; and ii. a plurality of inorganic material in a weight range of 0.1-30% with respect to the porous composite, wherein, the water-absorbing polymer matrix is impregnated with and is bound to the plurality of inorganic material.

2. The composite as claimed in claim 1, wherein the anti-microbial agent with the quaternary ammonium group is a compound of Formula II:

Formula II wherein,

R’ and R” are independently selected from hydrogen or Ci-6 alkyl;

R3 is selected from hydrogen, C1-6 alkyl, or C2-6 alkenyl;

X is selected from -O-, -N-, or -S-;

R4 is selected from hydrogen, Ci-30 alkyl, C2-30 alkenyl, or

wherein, Y is selected from -NH- or -O-;

3. The composite as claimed in any one of the claims 1 or 2, wherein the polymer matrix is a cross-linked polymerized product of: a. a functionalized compound; b. a monomer of a compound of Formula I:

Formula I wherein,

Ri is selected from hydrogen, Ci-6 alkyl, or C2-6 alkenyl;

R2 is selected from hydrogen, -CO-C1-6 alkyl, or -CO-C2-6 alkenyl; n' is an integer selected from 1 to 500; and c. theanti-microbial agent of a compound of Formula II:

Formula II wherein,

R’ and R” are independently selected from hydrogen or C1-6 alkyl;

R3 is selected from hydrogen, C1-6 alkyl, or C2-6 alkenyl;

X is selected from -O-, -N-, or -S-;

R4 is selected from hydrogen, Ci-30 alkyl, C2-30 alkenyl, or

wherein Y is selected from -NH- or -O-;

4. The composite as claimed in claim 3, wherein the functionalized compound comprises acrylate functionalized dextran, methacrylate functionalized dextran, acrylate functionalized cellulose, methacrylate functionalized cellulose, acrylate functionalized alginate, methacrylate functionalized alginate, acrylate functionalized chitosan, methacrylate functionalized chitosan, acrylate functionalized gelatin, methacrylate functionalized gelatin, or combinations thereof.

5. The composite as claimed in claim 4, wherein the functionalized compound is of Formula A:

Formula A, wherein,

6. The composite as claimed in claim 5, wherein the functionalized compound has a degree of substitution in a range of 1% to 50%.

7. The composite as claimed in claim 1, wherein the plurality of inorganic material is selected from SBA-15, MCM-41, laponite, or combinations thereof.

8. The composite as claimed in claim 3, wherein the polymer matrix cross-linked with the functionalized compound is in a weight range of 1-60%, the monomer of the compound of Formula I is in a weight range of 1-60%, and the antimicrobial agent with a quaternary ammonium group of Formula II is in a weight range of 1-60% in the presence of the plurality of inorganic material is in a weight range of 1-50%, with respect to the total weight of the porous composite.

9. The composite as claimed in claim 8, wherein the polymer matrix is cross-linked by photocuring, or thermal curing.

10. The composite as claimed in claim 1, wherein the composite has a pore size in a range of 1 pm to 100 pm and a specific surface area in a range of 1 m²/kg to 50

11. The composite as claimed in claim 1, wherein the composite possesses both hemostasis and antimicrobial property.

12. The composite as claimed in claim 1, optionally comprises at least one therapeutic agent selected from the group consisting of antimicrobial agents, antiinflammatory agents, wound healing promoting agents, anti-oxidant agents, antibiotic agents, antiseptics, antiparasitics, antifungal agents, and combinations thereof.

13. A process for preparation of hemostatic porous composite as claimed in any one of the claims 1 to 12 , the process comprising: a. mixing the functionalized compound, the monomer of the compound of Formula I, and the anti-microbial agent of a compound of Formula II with the plurality of inorganic material to obtain a reaction mixture; b. curing the reaction mixture to obtain a crosslinked polymerized product; and c. lowering a temperature followed by lyophilizing the crosslinked polymerized product to obtain a hemostatic porous composite.

14. The process as claimed in claim 13, wherein the reaction mixture is stirred for 30 min to 24 h at a temperature in a range of 4 °C to 50 °C in presence of a solvent.

15. The process as claimed in claim 13, wherein the lowering the temperature is carried out in a presence of liquid nitrogen for a time period of 5 to 60 mins.

16. The process as claimed in claim 13, wherein the curing is carried out by photocuring in presence of a photo -initiator selected from 2-Methyl-l-[4- (methylthio)phenyl] -2-(4-morpholinyl)- 1 -propanone, or 2-Hydroxy- 1 - [4-(2- hydroxyethoxy)-phenyl]-2-methyl-l -propanone; or by thermal curing using ammonium persulphate (APS) or N,N,N',N'-Tetramethylethylenediamine (TEMED).

17. The process as claimed in claim 13, wherein the functionalized compound is in a concentration range of 1 mg/mL to 100 mg/mL, the monomer of the compound of Formula I is in a concentration range of 1 mg/mL to 100 mg/mL, and the antimicrobial agent of a compound of Formula II is in a concentration range of 1 mg/mL to 100 mg/mL, and the plurality of inorganic material is in a concentration range of 1 mg/mL to 100 mg/mL, in the hemostatic porous composite.

18. A method for treating an injury in a subject, the method comprising: administering the hemostatic porous composite as claimed in claim 1 to an injury site; and allowing the hemostatic porous composite to swell and seal at the injury site.

19. The method as claimed in claim 18, wherein the composite imparts hemostasis and antimicrobial property at the injury site. 0. The method as claimed in claim 18, wherein the composite swells in contact with a fluid and has swelling ratio in a range of 1.5 to 50 times compared to initial size. 1. The method as claimed in claim 18, wherein the injury is selected from a wound, a hemorrhage, a hematoma, or a bleeding tissue. 2. A kit, comprising: the hemostatic porous composite as claimed in claim 1; and instructions for using the hemostatic porous composite. 3. A pharmaceutical composition comprising the hemostatic porous composite as claimed in claim 1 together with a pharmaceutically acceptable carrier, optionally in combination with other therapeutic agent. 4. The composition as claimed in claim 23, wherein the therapeutic agent is selected from antimicrobial agents, anti-inflammatory agents, wound healing promoting agents, anti-oxidant agents, antibiotic agents, antiseptics, antiparasitics, antifungal agents or combinations thereof. 5. An article comprising hemostatic porous composite as claimed in claim 1.