Wound dressing

Abstract

The present invention relates to a wound dressing material containing a modified polymer, wherein the modified polymer has the formula (I): 【Chemical 1】 JPEG2026509525000066.jpg35170 (where X is a covalent bond or a linker, and R 1 is, independently for each occurrence, halo, C 1~3 alkyl, C 2~3 alkenyl, C 1~3 haloalkyl, -O-C 1~3 alkyl, -O-C 1~3 haloalkyl and CN, and n is 0, 1, 2 or 3). It relates to a wound dressing material comprising a polymer backbone covalently bonded to a compound of. Similarly, a method of making a wound dressing material and the use of such a wound dressing material are provided. The present invention also relates to a method for detecting the presence of bacteria in a wound, comprising the step of applying the above-mentioned wound dressing material to the wound, and a method of forming a modified polysaccharide, wherein the polysaccharide, when it is part of a wound dressing material, relates to a method of forming a modified polysaccharide capable of detecting bacteria in a wound.

Description

[Technical Field]

[0001] The present invention relates to a wound dressing material comprising a modified polymer useful for detecting the presence of bacteria in a wound. Similarly, methods for preparing a wound dressing material and the use of such a wound dressing material are provided. The present invention also relates to a method for detecting the presence of bacteria in a wound, comprising the step of applying the above-described wound dressing material to a wound, and a method for forming a modified polysaccharide, wherein the polysaccharide is part of a wound dressing material, and the method for forming the modified polysaccharide capable of detecting bacteria in a wound. [Background technology]

[0002] A chronic wound is defined as a wound that does not heal through the usual sequence of stages and often persists for eight weeks without showing standard signs of healing (What Are the Treatment Options for Chronic Wounds?; Institute for Quality and Efficiency in Health Care (IQWiG), 2018). In fact, people with chronic wounds may remain with them for several months. The risk of developing chronic wounds increases with age (Yao, Z.; Niu, J.; Cheng, B. Prevalence of Chronic Skin Wounds and Their Risk Factors in an Inpatient Hospital Setting in Northern China. Advances in Skin & Wound Care 2020, 33 (9), 1-10) and health conditions such as diabetes (Greenhalgh, DG Wound Healing and Diabetes Mellitus. Clin Plast Surg 2003, 30 (1), 37-45) or cancer (Sundaram, GM; Quah, S.; Sampath, P. Cancer: The Dark Side of Wound Healing. The FEBS Journal 2018, 285 (24), 4516-4534). Treating chronic wounds is more complex and costly than treating ordinary wounds, and this can be further complicated by the increased risk of bacterial colonization, which delays healing, and the development of antimicrobial resistance in pathogenic strains. Antimicrobial resistance occurs when certain drugs that were previously inhibitory become ineffective against a microorganism.

[0003] The progression of wound infection can be divided into three stages: contamination of an open wound with non-replicating microorganisms, colonization of replicated strains, and infection where the increase in pathogen numbers affects the wound healing process and invasive cells migrate to deeper parts of the wound, causing inflammation and potential tissue damage (Haesler, E.; Ousey, K. Evolution of the Wound Infection Continuum. Wounds International 2018, 9 (4), 6-10; Li, S.; Renick, P.; Senkowsky, J.; Nair, A.; Tang, L. Diagnostics for Wound Infections. Adv Wound Care (New Rochelle) 2021, 10 (6), 317-32). Most wound infections are caused by bacteria, most commonly Staphylococcus aureus, Escherichia coli, Enterococcus spp., or Pseudomonas aeruginosa (Bowler, PG; Duerden, BI; Armstrong, DG Wound Microbiology and Associated Approaches to Wound Management. Clin Microbiol Rev 2001, 14 (2), 244-269; Saeed, MA; Haque, A.; Ali, A.; Mohsin, M.; Bashir, S.; Tariq, A.; Afzal, A.; Iftikhar, T.; Sarwar, Y. A Profile of Drug Resistance Genes and Integrons in E. Coli Causing Surgical Wound Infections in the Faisalabad Region of Pakistan. J Antibiot 2009, 62 (6), 319-323).

[0004] While the diagnosis of wound infection is usually based on clinical features or patient-reported symptoms, wound swab culture is increasingly being employed because it is a reliable and inexpensive means of pathogen identification (Hsu, J.-T.; Chen, Y.-W.; Ho, T.-W.; Tai, H.-C.; Wu, J.-M.; Sun, H.-Y.; Hung, C.-S.; Zeng, Y.-C.; Kuo, S.-Y.; Lai, F. Chronic Wound Assessment and Infection Detection Method. BMC Med Inform Decis Mak 2019, 19, 99). However, ensuring that samples are free from external contamination to minimize false positive results requires time and skill, which adds to the workload when treating wounds. Furthermore, sending samples to specialized laboratories, which may require culturing over several days, means that obtaining results takes a considerable amount of time. This means there is a greater chance that some kind of infection will progress considerably, making treatment even more difficult. In fact, it is unfortunately true that in some cases chronic wounds become very difficult to treat with antibiotics, and in order to save the patient's life, tissue excision or even amputation of the infected limb becomes necessary.

[0005] Therefore, there is a need for a timely diagnostic system that can be used to determine whether bacterial colonization is occurring in a point-of-care setting.

[0006] Certain enzyme substrates can evoke a chromogenic or fluorescent response in the presence of bacteria through enzymatic hydrolysis. These enzyme substrates may be specific to a narrower range of bacteria (e.g., substrates for L-alanylaminopeptidase cleaved by Gram-negative strains). However, some enzyme targets (e.g., L-leucylaminopeptidase) are thought to be common to all bacteria and can provide information about the presence of common bacteria (Colloms, SD Chapter 333 - Leucyl Aminopeptidase PepA. In Handbook of Proteolytic Enzymes (third Edition); Rawlings, ND, Salvesen, G., Eds.; Academic Press, 2013; pp 1484-1492). M. Cellier, et al., Bioorg. Med. Chem., 22, 2014, 5249-5269 disclose a series of chromogenic aminopeptidase substrates for the detection and identification of clinically important microorganisms.

[0007] An objective of certain embodiments of the present invention is to provide a convenient system that enables the detection of the presence of bacteria in a wound by, for example, causing a color change in a wound dressing. When removing the dressing from the patient, this would allow an observer to observe the distinguishable color on the surface of the dressing. [Overview of the project]

[0008] According to a first aspect of the present invention, a wound dressing material comprising a modified polymer, wherein the modified polymer is of formula (I): [ka] (In the formula, X is a covalent bond or linker, R 1 Each appearance is independent of the others, halo, C 1~3 Alkyl, C 2~3 Alkenil, C 1~3Haloalkyl, -OC 1~3 Alkyl, -OC 1~3 Selected from haloalkyl and CN, n is 0, 1, 2, or 3. [ka] (This indicates a bond point to the polymer backbone.) A wound dressing is provided which contains a polymer skeleton covalently bonded to the compound.

[0009] The covalent bonding of the compound of formula (I) to the polymer backbone results in a single-component modified polymer that can come into contact with the wound surface.

[0010] According to a second aspect of the present invention, a sterile package is provided comprising a wound dressing material according to a first aspect of the present invention. Advantageously, by providing the wound dressing material in a sterile package, the risk of the wound dressing material containing bacteria before application to the wound is minimized.

[0011] According to a third aspect of the present invention, the use of a wound dressing material according to the first aspect of the present invention is provided for detecting the presence of bacteria.

[0012] A fourth aspect of the present invention provides a method for detecting the presence of bacteria in a wound, comprising the steps of: applying a wound dressing material according to the first aspect of the present invention to the wound; removing the wound dressing material from the wound; and observing the coloration of the wound dressing material if any coloration is present.

[0013] According to a fifth aspect of the present invention, formula (III): [ka] (In the formula, X 1 is a covalent bond or linker, X 2is selected from NH, O and S, W is a monosaccharide residue, R 1 is, independently for each occurrence, halo, C 1~3 alkyl, C 2~3 alkenyl, C 1~3 haloalkyl, -O-C 1~3 alkyl, -O-C 1~3 haloalkyl and CN, n is 0, 1, 2 or 3), A method for forming a modified polysaccharide comprising a modified monosaccharide residue of), a) contacting an acidic polysaccharide with a compound of formula (IV):

Chemical formula

[0014] A sixth aspect provides a modified polysaccharide obtainable by the method of the fifth aspect or obtained by this method.

[0015] According to a seventh aspect of the present invention, there is provided a method for forming a wound dressing material comprising a modified polysaccharide, comprising the step of forming a modified polysaccharide by the method of the fifth aspect.

[0016] An eighth aspect of the present invention provides a wound dressing material obtainable by the method of the seventh aspect or obtained by this method.

[0017] According to a ninth aspect of the present invention, a method for forming a packaged wound dressing, i) The step of forming a wound dressing by the method of the seventh embodiment, ii) The step of packaging the wound dressing material to form the packaged wound dressing material, iii) Depending on the case, the step of sterilizing the packaged wound dressing and A method is provided that includes this.

[0018] A tenth aspect of the present invention provides a packaged wound dressing material that can be obtained by the method of the ninth aspect, or obtained by the method (optionally sterilized).

[0019] Embodiments of the present invention are further described herein with reference to the accompanying drawings. [Brief explanation of the drawing]

[0020] [Figure 1] This diagram shows the materials of alginate wound dressings: unmodified (left), and modified with compound 10 (center left) or compound 8 (center right and right). [Figure 2] This figure shows the materials of modified alginate wound dressings treated with EC 3a after overnight incubation. Left plate: top view; Right plate: bottom view. [Figure 3] This figure shows the materials of modified alginate wound dressings treated with EC 3b after overnight incubation. Left plate: top view; Right plate: bottom view. [Figure 4] This figure shows the materials of modified alginate wound dressings treated with PSA 3a after overnight incubation. Left plate: top view; Right plate: bottom view. [Figure 5]This figure shows the materials of modified alginate wound dressings treated with PSA 3b after overnight incubation. Left plate: top view; Right plate: bottom view. [Figure 6] This figure shows the materials of modified alginate wound dressings treated with SA 3a after overnight incubation. Left plate: top view; Right plate: bottom view. [Figure 7] This figure shows the materials of modified alginate wound dressings treated with SA at 3b after overnight incubation. Left plate: top view; Right plate: bottom view. [Modes for carrying out the invention]

[0021] Throughout this description and claims, the words “comprise” and “contain,” and their variations, mean “including, but not limited to,” and they are not intended to exclude (and do not exclude) other parts, additives, components, integers, or steps. Throughout this description and claims, the singular includes the plural unless the context specifically requires otherwise. In particular, where the indefinite article is used, this specification should be understood to intend both the singular and the plural unless the context specifically requires otherwise.

[0022] Features, integers, properties, compounds, chemical parts, or groups described in connection with specific aspects, embodiments, or examples of the present invention should be understood to be applicable to any other aspects, embodiments, or examples described herein, provided that they are not incompatible. All features disclosed herein (including any appended claims, abstract, and drawings) and / or all steps of any method or process disclosed herein may be combined in any way, except in combinations where at least some of such features and / or steps are mutually exclusive. The present invention is not limited to the details of any of the aforementioned embodiments. The present invention extends to any novel one or any novel combination of features disclosed herein (including any appended claims, abstract, and drawings), or any novel one or any novel combination of any steps of any method or process disclosed herein.

[0023] The reader's attention is directed to all documents and papers filed concurrently with or prior to this application and readily accessible in the public forum, all of which are incorporated herein by reference.

[0024] definition The term "linker" is intended to encompass any organic divalent radical. Examples of such organic divalent radicals include alkylenes (e.g., C1-C12). 12 Alkylene), PEG linker (e.g., -(OCH2CH2) m -or-(CH2CH2O) m - and m is an integer from 1 to approximately 100) and / or contains at least one of -S-, -SO2- or 1,2,3-triazolylene.

[0025] The term "polymer skeleton" is intended to encompass (unmodified) polymers that are converted into modified polymers. In other words, a "polymer skeleton" is a polymer that is covalently bonded to a compound of formula (I) in order to form a modified polymer. Suitable polymer skeletons for wound dressings may include synthetic polymers such as polyurethane, or polysaccharides such as cellulose, carboxymethylcellulose, carboxymethyl starch, pectin, alginic acid, carrageenan, heparin, gellan gum, agarose, hyaluronic acid, chitosan, or pharmaceutically acceptable salts thereof.

[0026] Biopolymers are all natural polymers, such as proteins, polynucleotides, and polysaccharides, that are formed by living organisms.

[0027] A polysaccharide is a polymer chain (often branched) containing multiple monosaccharide units linked together by glycosidic bonds. Naturally occurring polysaccharides can be found, for example, in plants, animals, microorganisms, and algae. However, the term “polysaccharide” is also intended to include synthetic polysaccharides or synthetically modified polysaccharides, such as sulfated, phosphorylated, or carboxymethylated polysaccharides.

[0028] Acidic polysaccharides are acidic groups, often -COOH (or -COO - ) and / or -OSO3H (or -OSO3 - This refers to a polysaccharide containing ). The acidic group may be in free form (i.e., -COOH and -OSO3H) or in salt form, i.e., -COO - and / or OSO3 - The group may also be in a form paired with a suitable counterion, such as sodium, potassium, calcium, or ammonium. The acidic group may also be naturally esterified, for example, with a methoxy group or an ethoxy group.

[0029] A "monosaccharide residue" (W) is a monosaccharide unit of an acidic polysaccharide that, upon modification, forms a modified monosaccharide residue of formula (III). For example, if W is guluronic acid, the modified monosaccharide residue of formula (III) has the following structure: [ka] It can have.

[0030] The "amino acids" (bound to the rest of the enzyme substrate by amide bonds) are typically protein-constituting amino acids, namely alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, selenocysteine, or pyrrolicin. The amino acids may also be α-amino acids, such as α-leucine, or β-amino acids, such as β-alanine. Usually, the amino acids are L-amino acids.

[0031] The term "pendant group" (or side-chain group) refers to a side branch from a chain that is neither an oligomer nor a polymer. For example, a phenyl group is a pendant group on a polystyrene chain.

[0032] Semi-closed dressings (semi-permeable dressings) protect wounds from external fluids while allowing the wound to "breathe" (air can penetrate and exit). Closed dressings are impermeable to both air and water.

[0033] The wound dressing material of the present invention makes it possible to indicate the presence of bacteria in a wound by coloring (color change) on the surface of the wound dressing material. The presence of a chromophore causes the coloring. The chromophore may be a blue chromophore that causes blue coloring (blue light that typically has wavelengths between 450 and 495 nm). Therefore, a blue chromophore typically absorbs light with wavelengths outside the range of 450 to 495 nm.

[0034] Term C m~n This refers to a group having m to n carbon atoms.

[0035] The term "alkyl" refers to a monovalent linear or branched saturated hydrocarbon chain. For example, C 1~3 Alkyl can refer to methyl, ethyl, n-propyl, or isopropyl.

[0036] The term "alkylene" refers to a divalent linear or branched saturated hydrocarbon chain. For example, C 1~8 Alkylene can refer to methylene, ethylene, n-propylene, isopropylene, n-butylene, sec-butylene, tert-butylene, n-pentylene, n-hexylene, or n-heptylene. The term "alkylene" is synonymous with the term "alkanediyl".

[0037] The term "alkenyl" refers to a monovalent linear or branched saturated hydrocarbon chain containing at least one double bond. The double bond can exist as an E isomer or a Z isomer. The double bond can be located at any possible position in the hydrocarbon chain. For example, "C 2~3 "Alkenyl" can refer to either ethenyl or propenyl.

[0038] The term "alkenylene" refers to a divalent linear or branched saturated hydrocarbon chain containing at least one double bond. The double bond can exist as an E isomer or a Z isomer. The double bond can be located at any possible position in the hydrocarbon chain. For example, "C 2~8"Alkenylene" can refer to etenylene, propenylene, butenylene, butadienylene, pentenylene, pentadienylene, hexenylene, hexadienylene, heptenylene, heptadienylene, or octenylene.

[0039] The term "alkynyl" refers to a monovalent linear or branched saturated hydrocarbon chain containing at least one triple bond. The triple bond can be located at any possible position in the hydrocarbon chain. For example, "C 2~4 "Alkinyl" can refer to ethynyl, propynyl, or butynyl.

[0040] The term "alkynylene" refers to a divalent linear or branched saturated hydrocarbon chain containing at least one triple bond. The triple bond can be located at any possible position in the hydrocarbon chain. For example, "C 2~8 "Alkinyl" can refer to ethynylene, propynylene, butynylene, pentynylene, hexynylene, heptynylene, or octinylene.

[0041] The term "haloalkyl" refers to an alkyl group (alkyl is defined above) in which one or more hydrogen atoms are substituted with a corresponding number of halogens. For example, the term "C 1~3 "Haloalkyl" is intended to encompass any saturated linear or branched alkyl moiety having 1 to 3 carbon atoms in which one or more hydrogen atoms are substituted by a corresponding number of halogen atoms. For example, "C 1~3 "Haloalkyl" includes, but is not limited to, trifluoromethyl (-CF3), 2,2,2-trifluoroethyl, 3-bromopropyl, and others.

[0042] The term "arirene" refers to any divalent aromatic carbocyclic ring system (i.e., a ring system containing 2(2n+1)π electrons). Arirenes may be monocyclic or polycyclic. An arirene group can have 6 to 12 carbon atoms in its ring system. Arirene groups are usually phenylene groups. Arirene groups may also be naphthylene groups or biphenylene groups.

[0043] The term "heteroarylene" refers to any 5-12 member divalent aromatic (i.e., a ring system containing 2(2n+1)π electrons) ring system containing 1-4 heteroatoms independently selected from O, S, and N (in other words, 1-4 atoms forming the ring system are selected from O, S, and N, and the rest are carbon). Heteroarylenes may be monocyclic or polycyclic. Heteroarylenes can be 6-membered heteroarylenes, such as pyridinylenes, in which the heteroaromatic ring is substituted with 1-3 (e.g., 1-2) nitrogen atoms.

[0044] When the term “substituted” is used herein in reference to a particular part, it means that one or more hydrogen atoms in that part, for example, one, two, or three, are independently replaced by the corresponding number of substituents described.

[0045] Naturally, it will be understood that substituents exist only in positions where they are chemically possible, and that those skilled in the art can determine, without undue effort (either experimentally or theoretically), whether a particular substitution is possible or not. For example, a hydrogen-free amino or hydroxyl group may be unstable when bonded to a carbon atom having an unsaturated (e.g., olefin) bond. Furthermore, it will be understood that the substituents described herein may themselves be substituted with any substituents, subject to the above-mentioned limitations on appropriate substitutions recognized by those skilled in the art.

[0046] When the arrangement of substituents on a particular group is determined by steric factors, the isomer with the lowest stereostructural energy may be preferred.

[0047] Where a compound, part, process, or product is described as "occasionally" having a certain feature, this disclosure includes both the case where such compound, part, process, or product has that feature and the case where such compound, part, process, or product does not have that feature. Thus, where a part is described as "occasionally substituted," this disclosure includes both the case where the part is not substituted and the case where the part is substituted.

[0048] When it is stated that two or more parts are selected "independently" or "each independently" from a list of atoms or groups, this means that the parts may be the same or different. Therefore, what each part is is independent of what one or more other parts are.

[0049] The term "pharmaceutically acceptable," as used herein, includes describing a compound, substance, composition, and / or dosage form that, within reasonable medical judgment, is suitable for use in contact with human or animal tissue without excessive toxicity, irritation, allergic reactions, or other problems or complications, and that meets a reasonable benefit / risk ratio. The term also includes being acceptable for both human and veterinary purposes.

[0050] The term “pharmaceutically acceptable salt” means a salt of an active compound prepared with a relatively non-toxic acid or base, depending on the specific substituents found in the compounds described herein. If a compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired base in neat or a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include salts or similar salts of sodium, potassium, calcium, ammonium, organic amino or magnesium. If a compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid in neat or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monocarbonate, phosphoric acid, monohydrogen-phosphoric acid, dihydrogen-phosphoric acid, sulfuric acid, monohydrogen-sulfuric acid, hydroiodic acid, or phosphorous acid, as well as salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. Similarly, salts of amino acids such as arginates, and salts of organic acids such as glucuronic acid or galacturonic acid are also included (see, for example, Berge et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present invention contain both basic and acidic functional groups, which enable the compound to be converted into a base addition salt or an acid addition salt.

[0051] The neutral form of the compound is preferably regenerated by contacting the salt with a base or acid and isolating the parent compound using conventional methods. The parent form of the compound differs from the various salt forms in terms of certain physical properties, such as solubility in polar solvents.

[0052] Certain compounds of the present invention may exist in non-solvated and solvated forms, including hydrated forms. Generally, the solvated forms are equivalent to the non-solvated forms and are included within the scope of the present invention. Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. Generally, for the uses envisioned by the present invention, all physical forms are intended to be equivalent and within the scope of the invention.

[0053] Certain compounds of the present invention have a chiral carbon atom (optical center) or a chiral double bond. Racemates, diastereomers, tautomers, geometric isomers, and individual isomers are included within the scope of the present invention. The compounds of the present invention do not include those known in the art to be too unstable to be synthesized and / or isolated.

[0054] The present invention also includes all pharmaceutically acceptable isotope-labeled compounds of formulas (I) to (V) (and isotope-labeled structural units of formulas (A), (Aa), (Ai) to (Aii) and (Bi) to (Biii)) in which one or more atoms are substituted by isotopes of the same element.

[0055] Examples of stable isotopes suitable for inclusion in the compound of the present invention include: 2 Hydrogen such as H, 11 C and 13 Carbon such as C, 15 Nitrogen such as N, as well as 17 O and 18 It contains oxygen isotopes such as O.

[0056] Isotope-labeled compounds can generally be prepared by conventional techniques known to those skilled in the art, or by methods similar to those described, using appropriate isotope-labeled reagents instead of previously used unlabeled reagents.

[0057] The abundance of isotopes can be determined using conventional analytical methods known to those skilled in the art, such as mass spectrometry and nuclear magnetic resonance spectroscopy.

[0058] Throughout this specification, these abbreviations have the following meanings:

[0059] JPEG2026509525000007.jpg112170

[0060] Wound dressing A first aspect of the present invention is a wound dressing comprising a modified polymer, wherein the modified polymer is of formula (I): [ka] (In the formula, X is a covalent bond or linker, R 1 Each appearance is independent of the others, halo, C 1~3 Alkyl, C 2~3 Alkenil, C 1~3 Haloalkyl, -OC 1~3 Alkyl, -OC 1~3 Selected from haloalkyl and CN, n is 0, 1, 2, or 3. [ka] (This indicates a bond point to the polymer backbone.) The present invention provides a wound dressing material containing a polymer skeleton covalently bonded to a compound.

[0061] Equation (I) X may be a covalent bond. However, usually X is a linker. The linker may have a molecular weight of less than 500 Da. The linker may have a molecular weight of less than 250 Da. The linker may have a molecular weight of less than 150 Da. The linker may have a molecular weight of less than 100 Da.

[0062] Linker is C 1~8 Alkylene, PEG linker (i.e., -(OCH2CH2)) m -or-(CH2CH2O) m-), -S-, -SO2- or 1,2,3-triazolylene can be included, where m is an integer selected from 1 to 20, C 1~8 Alkylenes are oxo, halo, and -OC. 1~3 Alkyl, -OC 1~3 Optionally, it is substituted with 1 to 4 substituents selected from haloalkyl and CN.

[0063] The modified polymer may be the product of a click reaction. Click reactions are well known in this field. Click reactions include thiol-ene reactions (see, e.g., AB Lowe, Polym. Chem., 2020, 1, 17-36), thiol-in reactions (see, e.g., R. Hoogenboom, Angew. Chem. Int. Ed., 2010, 49, 3415-7), SuFEx (see, e.g., J. Dong, L. Krasnova, MG Finn, KB Sharpless, Angew. Chem. Int. Ed., 2014, 53, 9430-9448), and azide-alkyne cycloaddition (see, e.g., F. Himo, T. Lovell, R. Hilgraf, VV Rostovtsev, L. Noodleman, KB Sharpless, VV Fokin, J. Am. Chem. Soc., 2005, 127). (See 210-216) When the modified polymer is synthesized by a thiol-ene reaction, the linker may contain -S-, e.g., -S-CH2CH2-. When the modified polymer is synthesized by a thiol-in reaction, the linker may contain -S-, e.g., -S-CH=CH-. When the modified polymer is synthesized by SuFEx, the linker may contain -SO2-. When the modified polymer is synthesized by azide-alkyne cycloaddition, the linker may contain 1,2,3-triazolylene, e.g., [ka] It may include.

[0064] Linker is -YZ 1 -L 1 -Z 2 - may be included, and Y is selected from -C(O)-, -S(O)2- and -S(O)-, Z 1 and Z 2 It is selected independently from O, S, and NH, and L 1 C 1~8 Alkylene, C 2~8 Alkenylene and C 2~8 Selected from alkynylene, L 1 If chemically possible, oxo, halo, -OC 1~3 Alkyl, -OC 1~3 Optionally substituted with 1 to 4 substituents selected from haloalkyl and CN. The linker is -YZ 1 -L 1 -Z 2 That's fine.

[0065] Y may be selected from -C(O)- and -S(O)2-. Y may also be -C(O)-. Z 1 It may also be NH. 2 It may also be NH. 1 and Z 2 Each of these may be NH. 1 C 1~8 Alkylene, for example, C 1~4 Alkylene may also be used. 1 This may be done without substitution.

[0066] R 1 Each appearance is independent of the halo and C. 1~3 It may be selected from alkyl groups. n may be 0, 1, or 2. n may be 0 or 1. Preferably, n is 0.

[0067] The compound of formula (I) is formula (Ia): [ka] (In the formula, Y, Z 1 , Z 2 , L1 and R 1 and n are as defined above) may be a compound of

[0068] The compound of formula (I) may be of formula (Ib):

Chemical formula

[0069] The compound of formula (I) may be selected from

Chemical formula

[0070] Polymer backbone Typically, the polymer backbone contains pendant groups. These pendant groups are reactive and enable (or enable) covalent coupling to the compound of formula (I). The pendant groups are oxo, =NR a , =NOR a , halo, -NO2, -CN, -N3, -NR a R a , -NR a S(O)2R a , -NR a C(O)R a , -NR a CONR a R a , -NR a CO2R a , -OR a , -SR a , -S(O)R a , -S(O)2R a , -S(O)2F, -SO3H, -OSO3H, -S(O)2NR a R a 、 -P(O)(OH)2, -B(OH)2, -CO2R a, -C(O)R a ,-CONR a R a , C 2~4 -alkyl, -C 2~4 -Alkenil, C 2~4 - Alkinyl and C 1~4 -Haloalkyl(R) a The group may be independently selected from H and C1-C4 alkyl groups (or pharmaceutically acceptable salts thereof) or may contain this group. The pendant group may be selected from -COOH, -OH, -NH2, -SH, -CH=CH2, -C≡CH, -S(O)2F, and -N3 or may contain this group.

[0071] Alternatively or further, the polymer backbone may contain heteroatoms, arylenes such as phenylene and heteroarylene, and / or unsaturated hydrocarbons, such as alkenylene or alkynylene, within the linear main chain of the polymer backbone. Such polymers include, for example, polyphenylene sulfide (PPS).

[0072] The polymer backbone may include biopolymers or synthetic polymers.

[0073] The polymer backbone may include synthetic polymers.

[0074] The polymer backbone may include polycarbonate, polyester, polyphthalamide, polyacrylate, polyarylate, polyamide-imide, polyimide, polyamide, polyolefin, polyurethane, chloroethylene, linear hydrocarbon polymer (e.g., polyethylene), poly(vinylnaphthalene), or copolymers thereof. The polymer backbone may also include polyurethane.

[0075] The polymer backbone may contain acrylonitrile butadiene styrene (ABS), polybutylene terephthalate (PBT), polyethylene terephthalate (PET), polyphenylene oxide, polyphenylene ether, polyphenylene sulfide, polystyrene, styrene-acrylonitrile resin (SAN), polyether ketone (PEEK), poly(methyl methacrylate) (PMMA), polyvinyl acetate, polysulfone, polyethersulfone, polyetherimide, polyvinyl chloride, polyethylene naphthalate, polypropylene, polyvinyltoluene, polyvinylpyrrolidone (PVP), or copolymers thereof.

[0076] The polymer backbone may contain a biopolymer. The biopolymer may be a polypeptide or a polysaccharide.

[0077] The polymer backbone may contain polypeptides. Typically, polypeptides are proteins or their (partial) hydrolysates. Proteins may be keratin, elastin, or collagen. The (partial) hydrolysate may be gelatin, which is sometimes referred to as hydrolyzed collagen. Proteins may be glycoproteins, such as fibrinogen. Proteins may be proteoglycans, such as heparan sulfate proteoglycans, chondroitin sulfate proteoglycans, or keratan sulfate proteoglycans.

[0078] The polymer backbone may contain a polysaccharide. The compound of formula (I) may be covalently bonded to the polysaccharide via an ester bond. Alternatively, the compound of formula (I) may be covalently bonded to the polysaccharide via an amide bond.

[0079] Polysaccharides may be glucans or modified forms thereof. Exemplary glucans include dextran, red algae starch, glycogen, pullulan, starch, cellulose, chrysolaminarin, curdlan, laminarin, lentinan, lichenin, oat beta-glucan, pleuran, and zymosan. Glucans may be cellulose or modified forms thereof. Polysaccharides may be modified polysaccharides, such as carboxymethylcellulose or carboxymethyl starch. Polysaccharides may be glycosaminoglycans.

[0080] The polysaccharide may be selected from the group consisting of cellulose, carboxymethylcellulose, carboxymethyl starch, pectin, alginic acid, carrageenan, heparin, gellan gum, agarose, hyaluronic acid, chitosan, or pharmaceutically acceptable salts thereof.

[0081] The polysaccharide may be or may contain an acidic polysaccharide. The acidic polysaccharide may contain acidic monosaccharide units such as uronic acid. Uronic acid includes glucuronic acid, galacturonic acid, iduronic acid, and mannuronic acid. The acidic polysaccharide may contain glucuronic acid and / or mannuronic acid monosaccharide units.

[0082] Acidic polysaccharides may also be glycosaminoglycans.

[0083] The acidic polysaccharide may be selected from the group consisting of heparin, chondroitin sulfate, dermatan sulfate, keratan sulfate, hyaluronic acid, gellan gum, pectin, heparan sulfate, alginic acid, carrageenan, carboxymethylcellulose, and carboxymethyl starch or pharmaceutically acceptable salts thereof.

[0084] The acidic polysaccharide may be alginic acid. The acidic polysaccharide may also be alginate, such as calcium alginate and / or sodium alginate.

[0085] The polymer backbone (and modified polymers) consists of individual structural units. More than 1% of the structural units may be covalently bonded to the compound of formula (I).

[0086] Modified polymers Modified polymers have structural units (A): [ka] (In the formula, W is a monosaccharide residue, and X and R are...) 1 (and n are as defined with respect to equation (I)) It may include.

[0087] Modified polymers have structural units (Aa): [ka] (In the formula, W is a monosaccharide residue, and X and R are...) 1 (and n are as defined with respect to equation (I)) It may include.

[0088] Modified polymers have structural units (Ai) and / or (Bi): [ka] (In the formula, X, R 1 (and n are as defined above) It may include.

[0089] Modified polymers have structural units (Aii) and / or (Bii): [ka] (In the formula, X, R 1 (and n are as defined above) It may include.

[0090] Modified polymers have structural units (Aiii) and / or (Biii): [ka] It may include.

[0091] More than 1% of the structural units of the modified polymer may be structural units (A), (Aa), (Ai), (Bi), (Aii), (Bii), (Aiii), or (Biii).

[0092] The modified polymer may also be a modified polymer matrix.

[0093] Wound dressing Wound dressings typically comprise an absorbent layer that is placed directly over the wound being treated. The absorbent layer is capable of absorbing wound exudate. Wound dressings may also comprise an absorbent layer that contains (for example, is formed from) a modified polymer. Enzyme substrates may be absorbed into the absorbent layer.

[0094] Wound dressings may be semi-occlusive or occlusive. For example, a wound dressing may comprise an absorbent layer and a (semi)occlusive layer located proximal to the absorbent layer so that the wound being treated can be protected from external fluids.

[0095] The wound dressing may contain an antimicrobial agent, such as medical-grade honey, iodine, silver, polyhexanide, dialkylcarbamoyl chloride, or bismuth tribromophenate. The antimicrobial agent may be silver, such as colloidal silver or silver nanoparticles.

[0096] The wound dressing may be a hydrophilic colloidal wound dressing. The wound dressing may further include an adhesive layer for adhering the wound dressing to the skin / tissue surrounding the wound. The wound dressing may have a pad portion (e.g., gauze) of material for taping the wound dressing to the wound.

[0097] The weight percentage of the modified polymer may be >50% by weight (of the total weight of the wound dressing). The weight percentage of the modified polymer may be >75% by weight. The weight percentage of the modified polymer may be >90% by weight, for example, 95% or 97.5% by weight.

[0098] The wound dressing material may, for example, be used to detect the presence of bacteria in a wound. The bacteria may be as defined below in relation to the third and fourth aspects of the present invention.

[0099] enzyme substrate In the embodiment, the wound dressing also comprises formula (II) [ka] It comprises an enzyme substrate or a pharmaceutically acceptable salt thereof, AA is an amino acid that is bound to the rest of the substrate via an amide bond. R 2 Each instance is independent of C 1~5 It is alkyl, p is 0, 1, 2, or 3. R 3 Each appearance is independent of the others, halo, C 1~3 Alkyl, C 2~3 Alkenil, C 1~3 Haloalkyl, -OC 1~3 Alkyl, -OC 1~3 Selected from haloalkyl and CN.

[0100] The enzyme substrate is given by formula (IIa) [ka] (In the formula, R 4 (where q is selected from H and amino acid side chains, q is 0 or 1, and R2, R3 and p are as defined above.) It may be a substrate of or a pharmaceutically acceptable salt thereof.

[0101] R 2 Each instance is independent of C 1~3 Alkyl may also be used. 2 Each instance is independent of C 1~2 Alkyl may also be used. 2 Each instance of it can be represented as Me. 2 Each instance of this can be represented as Et.

[0102] R 3 Each appearance is independent of the halo and C. 1~3 It may be selected from alkyl groups. p can be 0, 1, or 2. p can be 0 or 1. Preferably, p is 0.

[0103] R 4 R may be selected from H, Me, and i-Bu. 4 i-Bus is also acceptable.

[0104] The enzyme substrate is, [ka] You may choose from the following.

[0105] The enzyme substrate is, [ka] You may choose from the following.

[0106] The wound dressing may contain 1 to 100 mg of enzyme substrate per gram of modified polymer. The wound dressing may contain 1 to 80 mg of enzyme substrate per gram of modified polymer. The wound dressing may contain 1 to 60 mg of enzyme substrate per gram of modified polymer. The wound dressing may contain 10 to 60 mg of enzyme substrate per gram of modified polymer. The wound dressing may contain 1 to 40 mg of enzyme substrate per gram of modified polymer. The wound dressing may contain 20 to 80 mg of enzyme substrate per gram of modified polymer.

[0107] The wound dressing may contain 1 to 100 mg of enzyme substrate per gram of wound dressing. The wound dressing may contain 1 to 80 mg of enzyme substrate per gram of wound dressing. The wound dressing may contain 1 to 60 mg of enzyme substrate per gram of wound dressing. The wound dressing may contain 10 to 60 mg of enzyme substrate per gram of wound dressing. The wound dressing may contain 1 to 40 mg of enzyme substrate per gram of wound dressing. The wound dressing may contain 20 to 80 mg of enzyme substrate per gram of wound dressing.

[0108] The enzyme substrate is not covalently bonded to the polymer backbone. Typically, the enzyme substrate is absorbed into the wound dressing. The modified polymer is the modified polymer matrix, and the enzyme substrate may be dispersed throughout the modified polymer matrix.

[0109] Once a wound dressing is applied to a wound, the enzyme substrate diffuses freely within the wound. While we do not wish to be bound by theory, once the enzyme substrate diffuses into the wound and encounters bacteria that produce the appropriate enzyme, formula IIb; [ka] (In the formula, R2, R3, and p are as defined above with respect to formula II.) The compound is released by enzymatic hydrolysis of the enzyme substrate. The compound of formula IIb is thought to disperse within the wound dressing and return to the wound dressing, where it reacts with the compound of formula (I) to form a chromophore.

[0110] The presence of a chromophore causes a color change. This can be observed, for example, by visual inspection of the wound dressing when it is replaced, if the chromophore absorbs wavelengths in the visible range. Therefore, the wound dressing of the present invention makes it possible to indicate the presence of bacteria in a wound by a color change, for example, a blue color change. Since the color change is observed in-situ, treatment can be initiated quickly. Furthermore, the risk of external contamination is reduced compared to alternative bacterial detection systems that rely on swabs and remote analysis.

[0111] Similarly, a reacted wound dressing containing a modified polymer, wherein the modified polymer is of formula (V): [ka] The polymer skeleton is covalently bonded to the compound, and is used as a wound dressing, polymer skeleton, X, R 1 , R 2 , R 3 A wound dressing material is disclosed herein in which n and p are as defined above. The compound of formula (V) is formula (Va): [ka] It may also be a compound of the following.

[0112] Sterilization package A second aspect of the present invention provides a sterile package containing a wound dressing material according to the first aspect of the present invention. The sterile package may be a sterile pouch (or peel-off pack) made of plastic or paper containing the wound dressing material. The package may be sterilized by placing the package in a steam autoclave or by various methods such as dry heating, unsaturated chemical steam sterilization or gamma radiation sterilization.

[0113] How to use / detect In the uses and methods described herein, when referring to a "wound dressing according to a first aspect of the present invention" (or "wound dressing according to an eighth aspect of the present invention"), such wound dressings are included when removed from a sterile package according to a second (or tenth) aspect of the present invention. Thus, such uses and methods may also include a preliminary step of removing the wound dressing from the sterile package.

[0114] A third aspect of the present invention provides, for example, the use of a wound dressing material according to the first aspect (or eighth aspect) of the present invention for detecting the presence of bacteria in a wound.

[0115] A fourth aspect of the present invention provides a method for detecting the presence of bacteria in a wound, comprising the steps of: applying a wound dressing material according to the first aspect (or eighth aspect) of the present invention to the wound; removing the wound dressing material from the wound; and observing the coloration of the wound dressing material if any coloration is present. Coloration in the wound dressing material, for example, blue coloration, indicates the presence of bacteria.

[0116] The bacteria in the third or fourth aspect of the present invention may be a gram-negative strain of bacteria. The gram-negative strain of bacteria may be selected from Acinetobacter baumannii, Stenotrophomonas maltophilia, Escherichia coli, and Pseudomonas aeruginosa. The gram-negative strain of bacteria may be Escherichia coli or Pseudomonas aeruginosa.

[0117] The bacteria may be Gram-positive strains of bacteria. Gram-positive bacterial strains may include Pseudomonas aeruginosa, Enterococcus faecalis, Streptococcus pyogenes, and Staphylococcus aureus (e.g., methicillin-resistant Staphylococcus aureus (MRSA)). The Gram-positive bacterial strain may also be Staphylococcus aureus.

[0118] Bacteria can produce aminopeptidases such as L-alanylaminopeptidase or L-leucylaminopeptidase.

[0119] The wound may be a chronic wound. Typical chronic wounds include crusted wounds, necrotic wounds, or granulation wounds.

[0120] The wound is approximately 1 x 10 10 The wound may contain less than 1 x 10⁻¹⁰ bacteria. 8 The wound may contain less than 1 x 10⁻¹⁰ bacteria. 6 The wound may contain less than 1 x 10⁻¹⁰ bacteria. 4 The wound may contain less than 1 x 10⁻¹⁰ bacteria. 3 The wound may contain more than 1 x 10⁻¹⁰ bacteria. 4 The wound may contain more than 1 x 10⁻¹⁰ bacteria. 3 CFU ~ approximately 1 x 10 12 The wound may include a CFU (Chronic Fault Unit) of approximately 1 × 10⁻⁶. For example, the wound may be approximately 1 × 10⁻⁶. 4 CFU ~ approximately 1 x 10 10 This may include wounds within the range of CFU.

[0121] 1×10 6 The number of bacteria of 6 CFU is considered likely to progress towards causing infection in a wound. The exemplary wound dressing material of the present invention achieves a bacterial detection threshold below this level, i.e., this wound dressing material, when in contact with a wound that contains at least 1×10 6 CFU (or, for example, in the case of a detection threshold below said level, contains at least 1×10<C 5 CFU) for a required period of time, results in a detectable color change when in contact with the wound.

[0122] The method according to the fourth aspect of the present invention may include the step of applying a wound dressing material and removing the wound dressing material from the wound at least about 12 hours later. The method may include the step of applying a wound dressing material and removing the wound dressing material from the wound at least about 24 hours later. The method may include the step of applying a wound dressing material and removing the wound dressing material from the wound at least about 48 hours later. The method may include the step of applying a wound dressing material and removing the wound dressing material from the wound at least about 72 hours later. The method may include the step of observing any coloring on the surface of the wound dressing material while in contact with the wound. The coloring may be a blue coloring.

[0123] A further aspect of the present invention includes the use of a modified polymer as disclosed herein (e.g., a modified polymer of formula (I) or a modified polymer comprising any of structural units (A), (Aa), (Ai)-(Aii) and (Bi)-(Biii)) in a wound dressing material. [[ID=I8]]

[0124] Method for forming a modified polysaccharide The fifth aspect of the present invention is formula (III):

Chemical formula

Chemical formula

[0125] X 1 X 2 and X 3 X 1 may be a covalent bond. However, usually, X 1 is a linker. The linker may be -Y-Z 1 -L 1 -, Y is selected from -C(O)-, -S(O)2- and -S(O)-, Z 1 is selected from O, S and NH, L 1 is C 1~8 alkylene, C 2~8 alkenylene and C 2~8It is alkynylene, L 1 If chemically possible, oxo, halo, -OC 1~3 Alkyl, -OC 1~3 It is optionally substituted with 1 to 4 substituents selected from haloalkyl and CN. Y may be selected from -C(O)- and -S(O)2-. Y may also be -C(O)-. Z 1 L may be NH. 1 C 1~8 Alkylene, for example, C 1~4 It may be an alkylene. L1 may be ethylene. L1 may be unsubstituted.

[0126] X 2 X may be NH, 3 -NH2 may also be used.

[0127] R 1 and n R 1 Each appearance is independent of the halo and C. 1~3 It may be selected from alkyl groups. n may be 0, 1, or 2. n may be 0 or 1. Preferably, n is 0.

[0128] Step a) Step a) may include the step of suspending an acidic polysaccharide in a solvent to form a suspension. The solvent may be an aqueous solvent. For example, the solvent may include deionized water. The solvent may also include an organic solvent. Step a) may further include the step of adding a carboxyl activator to the suspension. Suitable carboxyl activators include carbodiimides and their salts. The carboxyl activator may be EDC, DCC, DIC or their hydrochloride salts. The carboxyl activator may be EDC. The carboxyl activator may be EDC·HCl.

[0129] Step a) may include stirring the suspension at room temperature. The suspension may be maintained at a pH of less than 7. The suspension may be maintained at a pH of less than 5. The suspension may be maintained at a pH of 2 to 4. The pH of the suspension may be adjusted by adding a suitable acid, such as HCl.

[0130] Step a) may include adding the compound of formula (IV) to the suspension to form a mixture. Alternatively, the compound of formula (IV) may be first dissolved in a suitable organic solvent such as DMSO or DMF before being added to the suspension.

[0131] Step a) may include stirring the mixture at room temperature. The mixture may be stirred for 24 hours or longer. The mixture may be stirred for 24 to 72 hours. The mixture may be maintained at a pH of less than 7. The mixture may be maintained at a pH of less than 5. The mixture may be maintained at a pH of 2 to 4. The pH of the mixture may be adjusted by adding a suitable acid, such as HCl.

[0132] Step aa) The method may further include step aa) isolating the modified polysaccharide. Step aa) may include filtering the mixture to obtain a filtrate containing the modified polysaccharide. The filtrate may be washed with an organic solvent, such as ethanol. The filtrate may be resuspended in an organic solvent, such as ethanol, stirred, refiltered, and washed again with an organic solvent, such as ethanol. This process may be repeated until the compound of formula (IV) is no longer detected in the filtrate (e.g., by TLC or HPLC evaluation). Step aa) may further include drying the modified polysaccharide for 3-4 hours, for example, in a vacuum oven.

[0133] Modified polysaccharides (C) and (E) This method uses modified polysaccharides (C): [ka] A method of forming, comprising contacting an acidic polysaccharide with compound (D):

Chem.

[0134] Alternatively, the method is a method of forming a modified polysaccharide (E):

Chem.

Chem.

[0135] Acidic polysaccharide The acidic polysaccharide may contain acidic monosaccharide units such as uronic acids. Uronic acids include glucuronic acid, galacturonic acid, iduronic acid and mannuronic acid. The acidic polysaccharide may contain glucuronic acid and / or mannuronic acid monosaccharide units.

[0136] The acidic polysaccharide may be a glycosaminoglycan.

[0137] The acidic polysaccharide may be selected from the group consisting of heparin, chondroitin sulfate, dermatan sulfate, keratan sulfate, hyaluronic acid, gellan gum, pectin, heparan sulfate, alginic acid, carrageenan, carboxymethyl cellulose and carboxymethyl starch or pharmaceutically acceptable salts thereof.

[0138] The acidic polysaccharide may be alginic acid. The acidic polysaccharide may be an alginate, such as calcium alginate and / or sodium alginate.

[0139] W W may be a uronic acid. For example, W may be glucuronic acid, galacturonic acid, iduronic acid, or mannuronic acid. W may be glucuronic acid. W may be mannuronic acid.

[0140] Enzyme substrate deposition The method may further include step b): depositing the enzyme substrate of formula (II) onto a modified polysaccharide. The enzyme substrate of formula (II) may be as defined above with respect to the first aspect of the present invention.

[0141] Step b) may include contacting the modified polysaccharide with a solution, for example, an aqueous solution containing an enzyme substrate, to form a wet modified polysaccharide. The step of contacting the modified polysaccharide with a solution may include adding the solution dropwise to the modified polysaccharide to form a wet modified polysaccharide. Alternatively, the step of contacting the modified polysaccharide with a solution may include adding the modified polysaccharide to a container containing the solution. The wet modified polysaccharide may then be dried.

[0142] The solution may contain 100-5000 mg / mL of enzyme substrate. The solution may contain 100-4000 mg / mL of enzyme substrate. The solution may contain 200-2000 mg / mL of enzyme substrate.

[0143] Step b) may include a step of depositing 1 to 100 mg of enzyme substrate per gram of modified polysaccharide. Step b) may include a step of depositing 1 to 75 mg of enzyme substrate per gram of modified polysaccharide. Step b) may include a step of depositing 1 to 60 mg of enzyme substrate per gram of modified polysaccharide. Step b) may include a step of depositing 10 to 60 mg of enzyme substrate per gram of modified polysaccharide. Step b) may include a step of depositing 20 to 75 mg of enzyme substrate per gram of modified polysaccharide. [Examples]

[0144] General synthesis Those skilled in the art will recognize that the application of methods known in the art may be applied in the manufacture of wound dressings, compounds, modified polymers, etc., according to the present invention.

[0145] For example, those skilled in the art may use the following as guidelines: “Comprehensive Organic Transformations - A Guide to Functional Group Transformations”, RC Larock, Wiley-VCH (1999 or later edition); “March's Advanced Organic Chemistry - Reactions, Mechanisms and Structure”, MB Smith, J. March, Wiley (5th edition or later edition); “Advanced Organic Chemistry, Part B, Reactions and Synthesis”, FA Carey, RJ Sundberg, Kluwer Academic / Plenum Publications (2001 or later edition); “Organic Synthesis - The Disconnection Approach”, S Warren (Wiley) (1982 or later edition); “Designing Organic Syntheses” S Warren (Wiley) (1983 or later edition); “Heterocyclic Chemistry”, J. Joule (Wiley 2010 edition or later edition); (“Guidebook To Organic Synthesis” RK Mackie and DM Smith) You should be directly familiar with standard textbooks such as (Longman) (1982 or later editions) and the references within them.

[0146] Those skilled in the art are familiar with a certain range of strategies for synthesizing organic molecules, particularly heterocyclic molecules, which represent common general knowledge described in textbooks such as Warren “Organic Synthesis: The Disconnection Approach”; Mackie and Smith “Guidebook to Organic Chemistry”; and Clayden, Greeves, Warren, and Wothers “Organic Chemistry.” Such strategies include "click chemistry," which has been extensively discussed in numerous books and journals, including AB Lowe, Polym. Chem., 2020, 1,17-36; R. Hoogenboom, Angew. Chem. Int. Ed., 2010, 49, 3415-7; J. Dong, L. Krasnova, MG Finn, KB Sharpless, Angew. Chem. Int. Ed., 2014, 53, 9430-9448; and F. Himo, T. Lovell, R. Hilgraf, VV Rostovtsev, L. Noodleman, KB Sharpless, VV Fokin, J. Am. Chem. Soc., 2005, 127, 210-216.

[0147] Those skilled in the art will also be familiar with a certain range of strategies for synthesizing modified derivatives of known polymers. Examples of such syntheses are F. Ilhan, et al., J. Am. Chem. Soc., 2000, 122, 5895-5896; F. Ilhan, et al., Macromolecules, 1999, 32, 6159-6162; F. Ilhan, et al., Macromolecules, 2001, 34, 2597-2601; AK Boal, et al., Tetrahedron, 2002, 58, 765-770; RJ Thibault, et al., J. Am. Chem. Soc., 2002, 124, 15249-15254; RJ Thibault, et al., J. Am. Chem. Soc., 2003, 125, 11249-11252; and described in JB Carroll, et al., Lett. Org. Chem., 2004, 1, 227-230.

[0148] Polysaccharides, such as cellulose, may be modified as described in Ma, H., Hsiao, BS, Chu, B. (2016). Modified Cellulose in Drioli, E., Giorno, L. (eds) Encyclopedia of Membranes. Springer, Berlin, Heidelberg.

[0149] [ka]

[0150] A skilled chemist, with regard to the synthesis of a given target compound, will be trained in their judgment and skill regarding the most efficient sequence of reactions and, where necessary, will use protecting groups. This will depend, in particular, on factors such as the nature of other functional groups present in a particular substrate. Clearly, the type of chemistry involved will influence the selection of reagents used in the synthesis steps, the need and type of protecting groups used, and the sequence in which the protection / deprotection steps are carried out. These and other reaction parameters will be apparent to those skilled in the art by referring to the standard textbook and the examples presented herein.

[0151] Highly sensitive functional groups may need to be protected and deprotected during the synthesis of the compounds of the present invention. This can be achieved, for example, by conventional methods described in PGM Wuts, Greene's Protective Groups in Organic Synthesis, John Wiley & Sons, Inc., Michigan, 2014 and its references.

[0152] All commercially available chemical reagents and solvents were obtained from Sigma-Aldrich, Apollo Scientific, Alfa Aesar, Fluorochem, or Fischer Scientific and used without further purification. Melting points were recorded using a Reichart-Kofler hot-stage microscope and were not corrected. Infrared spectra were recorded using a Perkin Elmer Spectrum BX FT-IR spectrophotometer at 4000–600 cm⁻¹. -1 The NMR spectrum was recorded within the range. 1 For the H spectrum, at 300 MHz, or 13For 14C spectra, the results were obtained using a Bruker Ultrashield300 spectrometer at 75 MHz. High-resolution, high-precision mass measurements were collected by the EPSRC National Mass Spectrometry Facility at Swansea University. Low-resolution mass spectra were recorded using a Bruker Esquire3000plus analyzer with either a positive or negative ion electrospray source. Thin-layer chromatography was performed using Merck silica gel 60F. 254 The above procedure was performed, and the results were analyzed by UV lamp (254 nm and 265 nm) and / or by adding ninhydrin staining (amine, amide-containing). Fischer silica gel 60 (35-70 microns) was used for column chromatography. The samples were pre-absorbed onto silica 60 (35-70 microns). Flash column chromatography was performed using a CombiFlash® NextGen100 system with RediSep Gold® normal-phase silica for normal-phase chromatography and RediSep Gold® C18 for reverse-phase chromatography.

[0153] Synthesis of enzyme substrates The enzyme substrate of formula (IIa) can be prepared in accordance with the synthesis disclosed in M. Cellier, et al., Bioorg. Med. Chem., 22, 2014, 5249-5269;DE2646033A1;EP2292613A1; and US6653312B1 A. The enzyme substrate of formula (IIa) can also be prepared by the method described in Scheme 1, or by a method in accordance with the specific synthesis of enzyme substrate 3a or 3b, or by a similar method.

[0154] [ka] Scheme 1: Reaction conditions: (i) Dry THF, IBCF, NMM, room temperature, overnight, N2 atmosphere; (ii) Ether, HCl (2M), room temperature, 2 hours.

[0155] tert-butyl(1-{[4-(dimethylamino)phenyl]amino}-4-methyl-1-oxopentan-2-yl)carbamate(2a)

[0156] [ka]

[0157] Under an inert atmosphere, Boc-L-leucine (1.02 g, 4.41 mmol, 1.2 equivalents) was dissolved in 20 mL of anhydrous THF in an ice bath. To this solution, NMM (0.49 mL, 4.41 mmol, 1.2 equivalents) and IBCF (0.58 mL, 4.41 mmol, 1.2 equivalents) were added dropwise, and the mixture was stirred at 0°C for 30 minutes. Dimethyl-4-phenylenediamine (1a) (0.50 g, 3.67 mmol, 1.0 equivalent) was added to the reaction mixture, and the ice bath was removed after 30 minutes. The reaction product was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, and the crude product was resuspended in DCM, then extracted with 0.1 M aqueous citric acid solution (2 × 30 mL), and then with 10% NaHCO3 aqueous solution (3 × 30 mL). The organic layer was washed with water (30 mL) and brine (30 mL) and dehydrated with MgSO4. The solvent was evaporated under vacuum, and the crude product was purified by flash column chromatography (gradient solvent method with silica gel solid phase, petroleum ether, and RINKAN mobile phase) to obtain 2a as a white powder (0.62 g, 1.8 mmol, 59%). Melting point: 144 - 145°C; 1 H NMR (300 MHz, CDCl3): δ 8.12 (s, 1H, 6-H), 7.34 (d, J o = 9.1 Hz, 2H, 3-H), 6.66 (d, J o = 9.0 Hz, 2H, 2-H), 5.11 (d, J HH= 8.3 Hz, 1H, 12-H), 4.31 - 4.18 (m, 1H, 8-H), 2.89 (s, 6H, 5-H), 1.88 - 1.54 (m, 3H, 9-H, 10-H), 1.44 (s, 9H, 15-H), 0.96 (d, J HH = 6.4 Hz, 11-H), 0.95 (d, J HH = 6.2 Hz, 11'-H) ppm; 13 C NMR (75 MHz, CDCl3): δ 170.4 (7-C), 156.1 (13-C), 148.0 (4-C), 127.8 (1-C), 121.6 (3-C), 113.0 (2-C), 80.3 (14-C), 77.43, 77.20, 77.00, 76.58, 53.7 (8-C), 41.1 (9-C), 40.9 (5-C), 28.3 (15-C), 24.8 (10-C), 22.0 (11-C, 11'-C) ppm; IR (υ max / cm -1 ): 3327 (NH), 2956 (CH), 1685 (C=O), 1653 (C=O), 1519 (NH), 1249 (CN), 1165 (CO); LRMS (ESI) m / z: 350.2 (MH+); HRMS (ESI) m / z actual value: 350.2446 (MH+); C 19 H 31 Calculated values ​​for N3O3: 349.2365 and C 19 H 32 N3O3 + The calculated value is 350.2438.

[0158] tert-butyl(1-{[4-(diethylamino)phenyl]amino}-4-methyl-1-oxopentan-2-yl)carbamate(2b)

[0159] [ka]

[0160] Under an inert atmosphere, Boc-L-leucine (1.02 g, 4.41 mmol, 1.2 equivalents) was dissolved in 20 mL of anhydrous THF in an ice bath. To this solution, NMM (0.49 mL, 4.41 mmol, 1.2 equivalents) and IBCF (0.58 mL, 4.41 mmol, 1.2 equivalents) were added dropwise, and the mixture was stirred at 0°C for 30 minutes. Diethyl-4-phenylenediamine (1b) (0.50 g, 3.67 mmol, 1.0 equivalent) was added to the reaction mixture, and the ice bath was removed after 30 minutes. The reaction product was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, and the crude product was resuspended in DCM, then extracted with 0.1 M aqueous citric acid solution (2 × 30 mL), and then with 10% NaHCO3 aqueous solution (3 × 30 mL). The organic layer was washed with water (30 mL) and brine (30 mL) and dehydrated with MgSO4. The solvent was evaporated under vacuum, and the crude product was purified by flash column chromatography (gradient solvent method with silica gel solid phase, petroleum ether, and SiO mobile phase) to obtain 2b as a white powder (0.91 g, 2.7 mmol, 75%). Melting point: 130 - 132°C; 1 H NMR (300 MHz, CDCl3) δ 7.93 (s, 1H, 7-H), 7.31 (d, J o = 9.1 Hz, 2H, 3-H), 6.62 (d, J o = 9.1 Hz, 2H, 2-H), 5.02 (d, J HH = 8.3 Hz, 1H, 13-H), 4.24-4.18 (m, 1H, 9-H), 3.31 (q, J HH = 7.1 Hz, 4H, 5-H), 1.82-1.51 (m, 3H, 10-H, 11-H), 1.45 (s, 9H, 16-H), 1.12 (t, J HH = 7.0 Hz, 4H), 0.96 (d, J HH = 6.4 Hz, 12-H), 0.95 (d, J HH = 6.2 Hz, 12'-H) ppm; 13C NMR (75 MHz, CDCl3) δ 170.2 (8-C), 156.0 (14-C), 145.2 (4-C), 126.6 (1-C), 122.0 (3-C), 112.5 (2-C), 80.3 (15-C), 53.7 (9-C), 44.6 (5-C), 41.1 (11-C), 28.3 (16-C), 24.8 (10-C), 22.9 (12-C, 12'-C), 22.05, 12.5 (6-C) ppm; IR (υmax / cm-1): 3290 (NH), 2966 (CH), 1653 (C=O), 1514 (NH), 1249 (CN), 1165 (CO); LRMS (ESI) m / z: 378.3 (MH+); HRMS (ESI) m / z actual value: 378.2757 (MH+); C 21 H 35 Calculated values ​​for N3O3: 377.2678 and C 21 H 36 N3O3 + The calculated value is 378.2751.

[0161] (2S)-2-amino-N-[4-(dimethylamino)phenyl]-4-methylpentanamide hydrochloride (3a)

[0162] [ka]

[0163] N-Boc protective amide 2a (1.0 equivalent) was stirred in a 2M HCl solution in diethyl ether (10.0 equivalents) at room temperature for 2 hours. After the reaction was complete, the mixture was filtered and washed with diethyl ether. Melting point: 248°C (Literature value: 193 - 197°C) 11 ); 1 H NMR (300 MHz, MeOD): δ 7.89 (d, J o = 9.2 Hz, 2H, 3-H), 7.69 (d, J o= 9.2 Hz, 2H, 2-H), 4.16-4.11 (m, 1H, 8-H), 3.29 (s, 6H, 5-H), 1.88-1.72 (m, 3H, 10-H, 9-H), 1.042 (d, J HH = 6.3 Hz, 3H, 11-H), 1.039 (d, J HH = 6.3 Hz, 3H, 11'-H) ppm; 13 C NMR (75 MHz, MeOD): δ 169.6 (7-C), 140.9 (1-C), 139.9 (4-C), 122.6 (2-C), 122.5 (3-C), 53.9 (8-C), 47.3 (5-C), 41.7 (9-C), 25.6 (10-C), 23.3 (11'-C), 22.2 (11-C) ppm; IR (υ max / cm -1 ): 2956 (CH), 2870 (CH), 1693 (C=O), 1514 (NH), 1261 (CN), 1116 (CN); LRMS (ESI) m / z: 250.2 (MH+); HRMS (ESI) m / z actual value: 250.1917 (MH+); C 14 H 23 Calculated values ​​for N3O, 249.1841 and C 14 H 24 N3O + The calculated value is 250.1914.

[0164] (2S)-2-amino-N-[4-(diethylamino)phenyl]-4-methylpentanamide hydrochloride (3b)

[0165] [ka]

[0166] N-Boc protective amide 2b (1.0 equivalent) was stirred in a 2M HCl solution in diethyl ether (10.0 equivalents) at room temperature for 2 hours. After the reaction was complete, the mixture was filtered and washed with diethyl ether. Melting point: 245 - 248°C;1 H NMR (300 MHz, MeOD): δ 7.91 (d, J o = 9.1 Hz, 2H, 3-H), 7.62 (d, J o = 9.1 Hz, 2H, 2-H), 4.17 (t, J HH = 7.1 Hz, 1H, 9-H), 3.67 (q, J HH = 7.2 Hz, 4H, 5-H), 3.52 (q, J = 7.0 Hz, 4H), 1.16 (t, J HH = 7.2 Hz, 6H, 6-H) ppm; 13 C NMR (75 MHz, MeOD): δ 169.9 (8-C), 140.8 (1-C), 134.3 (4-C), 124.5 (2-C), 123.1 (3-C), 55.1 (5-C), 53.9 (9-C), 41.6 (10-C), 27.3, 25.5 (11-C), 23.3 (12'-C), 22.3 (12-C), 10.8 (6-C) ppm; IR (υ max / cm -1 ): 2956 (CH), 2877 (CH), 1693 (C=O), 1508 (NH), 1257 (CN); LRMS (ESI) m / z: 278.2 (MH+); HRMS (ESI) m / z actual value: 278.2231 (MH+); C 16 H 27 Calculated values ​​for N3O: 277.2154 and C 16 H 28 N3O + The calculated value is 278.2227.

[0167] Synthesis of α-naphthol derivatives α-naphthol derivatives (e.g., compounds of formula (IV)) can be prepared by methods conforming to or similar to the synthesis of α-naphthol derivatives 8 or 10.

[0168] 4-amino-1-(8-hydroxynaphthalene-1-yl)butan-1-one(8) The amide-type 1-naphthol derivative (8) was synthesized on a gram scale in total yield of 38% from 1,8-naphthalic anhydride (4) using inexpensive commercial starting materials and simple reaction conditions in accordance with Scheme 2.

[0169] [ka] Scheme 2: Preparation of amide-type scavenger 8. Reaction conditions: (i) pyridine, HONH2·HCl, p-TsCl, reflux, 2 hours; (ii) H2O / EtOH, NaOH, reflux, 1 hour; (iii-1) aqueous NaOH (0.5M), reflux, overnight; (iii-2) slowly add NaNO2 at less than 5°C; add concentrated H2SO4 until pH < 4, raise temperature to 85°C; (iv) THF, ethylenediamine, reflux conditions, 1 hour.

[0170] (1,3-Dioxobenzo[de]isoquinoline-2-yl)4-methylbenzenesulfonate (5)

[0171] [ka]

[0172] 10.00 g of naphthal anhydride (4) (50.0 mmol, 1 equivalent) and 3.54 g of hydroxylamine hydrochloride (50.0 mmol, 1 equivalent) were added to 60 mL of pyridine, and the mixture was refluxed for 1 hour. Next, heating was stopped, and 21.00 g of p-toluenesulfonyl chloride (100.0 mmol, 2 equivalents) was added in four portions over 15 minutes. Heating was resumed for another hour, after which the mixture was added to distilled water (250 mL), the precipitate was filtered, and washed with 0.1 M NaOH solution (3 × 100 mL) and water (100 mL) to obtain 5 (18.00 g, 48.5 mmol, 97%) as a yellow powder. Melting point: 228 - 229°C (Literature value: 228 - 229°C) 12 ); 1 H NMR (400 MHz; DMSO-d6): δ 8.55 (dd, 2H, J o= 8.24 Hz, J m = 0.92 Hz, Ar-H), 8.50 (dd, 2H, J o = 7.33 Hz, J m = 0.92 Hz, Ar-H), 7.97 - 7.89 (m, 4H, Ar-H), 7.53 (d, 2H, J o = 8.24 Hz, Ar-H), 2.48 (s, 3H, 12-H) ppm; 13 C NMR (75 MHz, DMSO-d6): δ 160.1 (C-7), 147.1 (Ar-C), 136.0 (Ar-C), 132.1 (Ar-C), 132.1 (Ar-C), 132.0 (Ar-C), 130.6 (Ar-C), 129.5 (Ar-C), 128.0 (Ar-C), 127.2 (Ar-C), 122.4 (Ar-C), 21.8 (12-C) ppm; IR (υ max / cm -1 ): 3098 (C-H), 3061 (C-H), 1699 (C=O), 1176 (S=O); LRMS (ESI) m / z: 368.1 (MH + ); HRMS (ESI) m / z measured value: 368.0588 (MH + ); C 19 H 13 NO5S calculated value, 367.0(514) and C 19 H 14 NO5S + calculated value, 368.0587.

[0173] (1H-Benz[cd]indol-2-one)(6)

[0174]

Chemical Structure

[0175] A mixture consisting of 17.00 g of compound 5 (46.0 mmol, 1 equivalent) and 10.00 g of NaOH (250.0 mmol, 5.5 equivalents) was stirred under reflux for 1 hour in a mixture of distilled water and ethanol (200 mL, 3:1). Heating was continued for another hour, during which time the ethanol was evaporated. The mixture was cooled and acidified with concentrated HCl (30 mL) to generate CO2 and precipitate the product. The crude product was filtered to obtain compound 6 (6.5 g, 38.2 mmol, 83%) as a yellow powder (6.5 g, 38.2 mmol, 83%). Melting point: 180°C (Literature value: 180 - 182°C) 13 ); 1 H NMR (400 MHz; DMSO-d6): δ10.80 (s, 1H, 9-H), 8.18 (d, 1H, J o = 7.79 Hz, Ar-H), 8.03 (d, 1H, J o = 6.87 Hz, Ar-H), 7.78 (dd, J o = 8.0 Hz, J o = 7.0 Hz, 1H, Ar-H), 7.61 (d, 1H, J o = 8.24 Hz, Ar-H), 7.48 (dd, J o = 8.4 Hz, J o = 7.0 Hz, 1H, Ar-H), 7.00 (d, 1H, J o = 6.87 Hz, Ar-H) ppm; 13 C NMR (75 MHz; DMSO-d6): δ 169.4 (10-C), 138.6 (Ar-C), 131.4 (Ar-C), 129.6 (Ar-C), 129.5 (Ar-C), 129.4 (Ar-C), 127.4 (Ar-C), 126.2 (Ar-C), 124.2 (Ar-C), 119.9 (Ar-C), 106.6 (Ar-C) ppm; IR (υ max / cm -1): 3067 (NH), 3089 (CH), 1716 (C=O), 1494 (NH); HRMS (ESI) m / z actual value: 170.0606 (MH+); LRMS (ESI) m / z: 170.1 (MH+); C 11 Calculated values ​​of H7NO, 169.0528 and C 11 H8NO + The calculated value is 170.0600.

[0176] 2-Oxatricyclo[6.3.1.0^{4,12}]dodeca-1(11),4,6,8(12),9-pentaen-3-one(7)

[0177] [ka]

[0178] 5.00 g of 6 (29.6 mmol, 1.2 equivalents) was dissolved in 200 mL of 0.5 M NaOH (aqueous solution), and the mixture was stirred overnight under reflux. The mixture was cooled using an ice bath and maintained below 5°C. 1.74 g of NaNO2 (25.00 mmol, 1 equivalent) was added to this solution and stirred for 10 minutes. In a conical flask, approximately 100 mL of water was prepared with concentrated H2SO4 to a pH of less than 2, and this solution was cooled in an ice bath to below 5°C. The reaction mixture was slowly added to the acidic solution under pH and temperature control until the addition was complete and the crude product crashed out of the solution. The resulting mixture was then slowly heated to 85°C to generate N2. The mixture was stirred at room temperature for 2 hours, after which the precipitate was filtered and washed with water. When the product was dried, compound 7 was obtained as a brown solid (3.80 g, 22.5 mmol, 76%). Melting point: 105 - 108°C (Literature value: 105 - 107.5°C) 14 ); 1 H NMR (300 MHz, DMSO-d6): δ 8.40 (d, J o = 8.2 Hz, 1H, Ar-H), 8.29 (d, Jo = 7.1 Hz, 1H, Ar-H), 7.96 (dd, J o = 8.0 Hz, J o = 7.2 Hz, 1H, Ar-H), 7.85 (d, J o = 8.5 Hz, 1H, Ar-H), 7.69 (dd, J o = 8.4 Hz, J o = 7.4 Hz, 1H, Ar-H), 7.37 (d, J o = 7.3 Hz, 1H, Ar-H) ppm; 13 C NMR (75 MHz, DMSO-d6): δ 167.0 (9-C), 149.8 (Ar-C), 133.1 (Ar-C), 130.6 (Ar-C), 130.0 (Ar-C), 129.9 (Ar-C), 128.8 (Ar-C), 127.2 (Ar-C), 121.6 (Ar-C), 120.7 (Ar-C), 106.6 (Ar-C) ppm; IR (υ max / cm -1 ): 3072 (C-H), 1774 (C=O), 1226 (C-O); LRMS (ESI) m / z: 171.0 (MH+); HRMS (ESI) m / z measured value: 171.0445 (MH+); C 11 H6O2 calculated value, 170.0368 and C 11 H7O2 + calculated value, 171.0441.[[ID=​​​​​​​​​​​​​​Ethylenediamine (1.60 mL, 23.91 mmol, 4.0 equivalents) and compound 7 (1.02 g, 5.98 mmol, 1.0 equivalent) were dissolved in 30 mL of anhydrous THF. This mixture was refluxed for 1 hour, filtered, washed with THF, and the resulting product was crystallized from methanol to obtain compound 8 as a pink powder (0.92 g, 4.00 mmol, 68%). Melting point: 216 - 217°C (Literature value: 200 - 201°C) 15 ); 1 H NMR (300 MHz, DMSO-d6): δ 8.15 (brs, 1H, 9-H), 7.83 (dd, J o = 8.2 Hz, J m = 1.3 Hz, 1H, Ar-H), 7.40 (dd, J o = 8.2 Hz, J o = 7.0 Hz, 1H, Ar-H), 7.36-7.20 (m, 3H, Ar-H), 6.82 (dd, J o = 8.8 Hz, J m = 1.4 Hz, 1H, Ar-H), 3.31 (t, 2H, J HH = 6.03 Hz, 12-H), 2.75 (t, 2H, J HH = 6.03 Hz, 13-H) ppm; 13 C NMR (75 MHz, DMSO-d6): δ 172.4 (C-10), 155.1 (Ar-C), 135.7 (Ar-C), 134.9 (Ar-C), 129.2 (Ar-C), 127.4 (Ar-C), 125.8 (Ar-C), 124.8 (Ar-C), 121.8 (Ar-C), 118.3 (Ar-C), 110.9 (Ar-C), 42.9 (12-C), 41.5 (13-C) ppm; IR (υ max / cm -1): 3263 (NH), 2970 (CH), 1639 (C=O), 1552 (NH), 1271 (CN), 1012 (CO); LRMS (ESI) m / z: 230.3 (MH+); HRMS (ESI) m / z actual value: 231.1131 (MH+); C 13 H 14 Calculated values ​​for N2O2: 230.1055 and C 13 H 15 N2O2 + The calculated value is 231.1128.

[0182] N-(2-aminoethyl)-8-hydroxynaphthalene-1-sulfonamide(10) The sulfonamide-type scavenger (10) was synthesized in a one-step reaction in 80% yield on a gram scale using naphthosultone (9) as the starting material, in accordance with Scheme 3. [ka] Scheme 3: Synthesis of sulfonamide molecule 10. Reaction conditions: (i): THF, ethylenediamine, reflux, 1 hour.

[0183] N-(2-aminoethyl)-8-hydroxynaphthalene-1-sulfonamide(10)

[0184] [ka]

[0185] Ethylenediamine (3.88 mmol) was added to a solution of naphthosultone (9) (0.97 mmol) in THF (4.95 mL), and the solution was refluxed for 1 hour. After removing the THF under reduced pressure, the product was precipitated by adding ethanol and filtered. A yellow solid (207 mg, 80%) was obtained. Melting point: 120°C; 1H-NMR (300 MHz; d6-DMSO) δ: 8.06 (1H, d, J = 7.35 Hz, CH-Ar), 7.98 (1H, d, J = 7.72 Hz, CH-Ar), 7.48-7.30 (3H, m, CH-Ar), 6.91 (1H, d, J = 6.78 Hz, CH-Ar), 2.95-2.87 (2H, m, CH2), 2.78-2.65 (2H, m, CH2); IR (υ max / cm -1 ): 3510.45w (NH), 3300 wide (OH), 2970s (CH2), 1519.91s (C=C), 1276.88s (SO2).

[0186] Modified polysaccharides The modified polysaccharides of the present invention can be prepared by a method conforming to or similar to the method described in Scheme 4.

[0187] [ka]

[0188] Wound dressing An exemplary wound dressing of the present invention can be prepared by a method conforming to or similar to the method described in Scheme 4. [ka] This forms a portion of the wound dressing material. The wound dressing material of the present invention can be prepared by a method in accordance with or similar to the method described in "Modification of ActivHeal® Alginate" below.

[0189] Modification of ActivHeal® alginate A sample of ACTIVHEAL® simply A (10 × 20 cm) non-adhesive foam was provided by Advanced Medical Solutions Ltd. Using scissors cleaned with ethanol, the wound dressing material was cut into squares of approximately 5-10 × 5-10 mm. A maximum amount of 0.20 g of the material was added to 10 mL of DI water, and the pH was adjusted to approximately 3 by adding 0.1 M HCl aqueous solution. This solution was stirred at room temperature for 10 minutes, and then 0.33 g of EDC·HCl (1.7 mmol) was added, and the pH was maintained at approximately 3 by adding 0.1 M HCl aqueous solution. This solution was stirred at room temperature for 10 minutes, and then 0.88 g of compound 8 dissolved in DMSO or DMF was added. This mixture was stirred at room temperature for 48 hours. The wound dressing material was filtered and washed with ethanol. To ensure complete removal of any unbound amides (8), the foam sample was resuspended in ethanol, stirred for 10 minutes, filtered, and repeated until amides (8) were undetectable in the filtrate (this could be maintained by TLC or HPLC evaluation). The sample was then dried in a vacuum oven for 3–4 hours.

[0190] Aqueous solutions of 3a or 3b were prepared as follows: The desired amount of enzyme substrate (3.0 mg for 300 mg / L, 6.0 mg for 600 mg / L, 9.0 mg for 900 mg / L, 15.0 mg for 1500 mg / L, and 30.0 mg for 3000 mg / L) was weighed using a balance with an error limit of ±0.3 mg to four decimal places, placed in a 10 mL volumetric flask, and adjusted to 10.0 mL with DI water. This solution was absorbed into the foam sample and dried overnight in a vacuum oven at 40°C.

[0191] microbiology All culture media and reagents used in the microbiological evaluation were obtained from either Fisher Scientific (Loughborough, UK) or Sigma Aldrich (Dorset, UK) and were of general laboratory grade unless otherwise specified in the text.

[0192] All bacterial strains used were produced by and published by the National Collection of Type Cultures. They are summarized in the table below.

[0193] JPEG2026509525000047.jpg53170

[0194] All cultures were maintained at -80°C using Microbank Cryobeads (Pro-Lab Diagnostics, CA). The cultures were then restored from the cryopreservation chamber to tryptic soy agar (TSA) working stock plates.

[0195] Bacterial strains were cultured in tripty soy broth (TSB) from a preservation solution using standard methods and incubated overnight at 37°C with shaking at 150 RPM. The overnight culture was centrifuged at 3000 × g for 5 minutes, the supernatant was discarded, and the pellet was resuspended in 10 mL of phosphate-buffered saline (PBS). 1-3 × 10 8 The bacterial inoculum was standardized by optical density at 600 nm using an Amersham Biosciences Ultrospec10 spectrophotometer (Amersham Biosciences, Amersham, UK) to contain between CFU / mL.

[0196] A lower concentration bacterial solution is prepared by diluting 500 μL of standard microbial solution in 4500 μL of PBS or sterile physiological saline, and 10 7 CFU / mL was obtained.

[0197] Microbiological evaluation of bacterial suspensions overnight To model fluid exudate from chronic wounds, wound dressings prepared according to the "Modification of ActivHeal® Alginate" described above were placed on agar plates, and test pathogens were rubbed onto the surface of the agar.

[0198] The enzyme substrate was tested at four different concentrations (300, 600, 900, and 3000 mg / L) using DI water as the solvent. These solutions were absorbed into wound dressing samples and dried overnight under vacuum at 40°C.

[0199] Two Gram-negative bacterial strains (Escherichia coli-EC and Pseudomonas aeruginosa-PSA) and a Gram-positive bacterium (Staphylococcus aureus-SA) were evaluated against dimethyl(3a) and diethyl(3b) substituents. Overnight cultures were rubbed directly from LB broth onto Columbia blood agar without measuring OD. Wound dressings containing absorbed enzyme substrates were placed on the agar and incubated overnight (see Figures 2-7).

[0200] The test results for N,N-dimethyl substrate 3a and N,N-diethyl substrate 3b are summarized in the table. In Table 2, the intensity of the observable color change is pale blue < light blue < blue < dark blue. The stronger the color change, the easier it is to identify the presence of bacteria. Both Gram-negative strains (EC and PSA) showed a blue color change with a 300 mg / L concentration of substrate 3b, but only SA showed a color change when a higher concentration (3000 mg / L) of substrate 3b was applied. The N,N-diethyl substrate (3b) produced a stronger color change than the dimethyl analog (3a).

[0201] It is important to note that applying a substrate at a very high concentration (in this case, 3000 mg / L) may lead to the diffusion of oxidation products of hydrolyzed aniline, which can cause the agar to turn brown.

[0202] JPEG2026509525000048.jpg69170

[0203] Microbiological evaluation of standardized bacterial suspensions Microbiological evaluation of the bacterial suspension overnight showed promising results. However, for desirable clinical applications, rapid evaluation of lower bacterial concentrations is essential. Three different solutions (300, 600, and 1500 mg / L) were prepared from substrates 3a and 3b in microfiltered DI water as the solvent. These solutions were absorbed into wound dressing samples and dried overnight under vacuum at 40°C.

[0204] Two Gram-negative bacterial strains (Escherichia coli-EC and Pseudomonas aeruginosa-PSA) and a Gram-positive bacterium (Staphylococcus aureus-SA) were evaluated. The bacteria were incubated overnight, centrifuged at 3000 × g for 5 minutes, the supernatant was discarded, and the residual pellet was resuspended in 10 mL of PBS. (1-3 × 10⁻⁶) 8 The inoculum was standardized by measuring the optical density at 600 nm to ensure a CFU / mL concentration. The standardized solution was further diluted to 1–3 × 10⁻⁶. 6 and 1-3 × 10 4 A suspension of CFU / mL was prepared. 1 × 10 6 CFU is considered an ideal threshold for a positive result because this concentration closely matches the level of colonization where progression and infection are likely to have occurred.

[0205] A 100 μL bacterial suspension was smeared onto an agar plate. A wound dressing sample was added to the top of the agar and swirled several times (to ensure the entire area was covered and to facilitate visual observation of the color change). The plate was incubated at 37°C, and results were recorded at 24, 48, and 72 hours. The results are shown in Tables 3-5. - indicates no color change, +- indicates a relatively recognizable blue change, + indicates a moderately strong blue change, and ++ indicates a strong blue change.

[0206] JPEG2026509525000049.jpg57170

[0207] JPEG2026509525000050.jpg58170

[0208] JPEG2026509525000051.jpg58170

[0209] After 24 hours of incubation, EC(10 8 CFU showed a weak color change in both substrates, but the blue color was barely visible.

[0210] After 48 hours of incubation, the EC and PSA strains, which are Gram-negative strains (10 8 Both CFUs, at higher substrate concentrations (900 or 1500 mg / L), produced a strong blue color change in the case of substrate 3b, while EC(10 6 CFU and 10 4 CFU showed a moderate color change at lower bacterial concentrations. Both EC and PSA, which are Gram-negative bacteria, showed a moderate color change at higher or lower bacterial concentrations (10) with N,N-diethyl substrate 3b (900 mg / L). 8 CFU, 10 6 CFU and 10 4 CFU produced a strong blue color change, while N,N-dimethyl substrate 3a produced a moderate or strong color change at higher substrate concentrations (900 or 1500 mg / L). The SA of the Gram-positive organisms evaluated showed a weak but distinguishable color change when the evaluated substrate (3a or 3b at 1500 mg / L) was present at the highest concentration.

[0211] 72 hours later, 10 8 , 10 6 or 10 4 Both Gram-negative strains EC and PSA in the CFU showed moderate to strong blue color changes at the tested substrate concentrations for both substrates 3a and 3b. The Gram-positive organism SA, which was evaluated, showed only a weak, almost invisible color change when the evaluated substrate (1500 mg / L of 3a or 3b) was present at its highest concentration.

Claims

1. A wound dressing comprising a modified polymer, wherein the modified polymer is of formula (I): 【Chemistry 1】 It contains a polymer backbone that is covalently bonded to the compound, X is a covalent bond or linker, R １ Each appearance is independent of the others, halo, C １～３ Alkyl, C ２～３ Alkenil, C １～３ Haloalkyl, C １～３ Alkoxy, C １～３ Selected from haloalkoxys and CN, n is 0, 1, 2, or 3. 【Chemistry 2】 This is a wound dressing material that shows a bonding point to the polymer backbone.

2. The aforementioned linker, C １～８ alkylene, -(OCH ２ CH ２ ), ｍ -(CH ２ CH ２ O), ｍ -S-, -SO ２ - or 1,2,3-triazolylene (m is an integer selected from 1 to 20, and C １～８ alkylene is optionally substituted by 1 to 4 substituents selected from oxo, halo, C １～３ alkoxy, C １～３ haloalkoxy and CN), and / or -Y-Z １ -L １ -Z ２ -(Y is -C(O)-, -S(O)) ２ - and -S(O)- are selected, Z １ and Z ２ It is selected independently from O, S, and NH, and L １ C １～８ Alkylene, C ２～８ Alkenylene and C ２～８ Selected from alkynylene, L １ Where chemically possible, oxo, halo, C １～３ Alkoxy, C １～３ (Optionally substituted with 1 to 4 substituents selected from haloalkoxys and CNs.) A wound dressing according to claim 1, comprising:

3. The compound of formula (I) is of formula (Ia): 【Transformation 3】 It is a compound of, Y is -C(O)-, -S(O) ２ - and -S(O)- are selected, Z １ and Z ２ It is selected independently from O, S, and NH. L １ C １～８ Alkylene, C ２～８ Alkenylene and C ２～８ It is alkynylene, L １ Where chemically possible, oxo, halo, C １～３ Alkoxy, C １～３ The wound dressing according to claim 1 or 2, optionally substituted with one to four substituents selected from haloalkoxys and CNs.

4. A wound dressing according to any one of claims 1 to 3, wherein n is 0.

5. The compound of formula (I) above, 【Chemistry 4】 A wound dressing according to any one of claims 1 to 4, selected from the above.

6. The polymer backbone is -COOH or -COO － A wound dressing material according to claim 5, comprising a base.

7. The wound dressing according to claim 5 or 6, wherein the polymer skeleton comprises a polysaccharide.

8. The wound dressing according to claim 7, wherein the compound of formula (I) is covalently bonded to the polysaccharide via an ester bond or an amide bond.

9. The wound dressing according to claim 7 or 8, wherein the polysaccharide comprises an acidic polysaccharide or a pharmaceutically acceptable salt thereof.

10. The wound dressing according to any one of claims 7 to 9, wherein the polysaccharide comprises alginic acid or a pharmaceutically acceptable salt thereof.

11. The modified polymer is a structural unit (A) and / or (B): 【Transformation 5】 A wound dressing according to claim 7, comprising:

12. The wound dressing material is of formula (II): 【Transformation 6】 Further comprising the enzyme substrate or a pharmaceutically acceptable salt thereof, AA is an amino acid that is bound to the rest of the substrate via an amide bond. R ２ Each instance is independent of C １～５ It is alkyl, p is 0, 1, 2, or 3. R ３ Each appearance is independent of the others, halo, C １～３ Alkyl, C ２～３ Alkenil, C １～３ Haloalkyl, C １～３ Alkoxy, C １～３ (Selected from haloalkoxy and CN) A wound dressing according to any one of claims 1 to 11.

13. The enzyme substrate is given by formula (IIa): 【Transformation 7】 A substrate or a pharmaceutically acceptable salt thereof, R ４ It is selected from H and amino acid side chains, The wound dressing material according to claim 12, wherein q is 0 or 1.

14. A wound dressing according to claim 12 or 13, wherein p is 0.

15. The aforementioned substrate, 【Transformation 8】 A wound dressing according to any one of claims 12 to 14, selected from the above.

16. The wound dressing according to any one of claims 12 to 15, wherein the enzyme substrate is absorbed into the wound dressing.

17. A sterile package comprising a wound dressing material according to any one of claims 1 to 16.

18. Use of the wound dressing material according to any one of claims 1 to 16 for detecting the presence of bacteria.

19. A method for detecting the presence of bacteria in a wound, comprising the steps of: applying a wound dressing material according to any one of claims 1 to 16 to the wound; removing the wound dressing material from the wound; and observing the coloration of the wound dressing material if it is present.

20. The use according to claim 18 or the method according to claim 19, wherein the bacterium is a gram-negative strain of bacteria.

21. The use or method according to claim 20, wherein the Gram-negative bacterial strain is Escherichia coli or Pseudomonas aeruginosa.

22. The use according to claim 18 or the method according to claim 19, wherein the bacterium is a Gram-positive bacterial strain.

23. The use or method according to claim 22, wherein the Gram-positive bacterial strain is Staphylococcus aureus.

24. Formula (III): 【Chemistry 9】 A method for forming a modified polysaccharide containing a modified monosaccharide residue, X １ is a covalent bond or linker, X ２ It is selected from NH, O, and S. W is a monosaccharide residue, R １ Each appearance is independent of the others, halo, C １～３ Alkyl, C ２～３ Alkenil, C １～３ Haloalkyl, C １～３ Alkoxy, C １～３ Selected from haloalkoxys and CN, n is 0, 1, 2, or 3. a) Acidic polysaccharide formula (IV): 【Chemistry 10】 The step of contacting the compound to form the modified polysaccharide is included. X ３ is, -NH ２ Selected from -OH and -SH, However, X ２ If X is NH, ３ Ha-NH ２ X ２ If X is O, ３ is -OH, and X ２ If S, then X ３ The method is -SH.

25. X １ This is a linker, and in some cases the linker is -Y-Z １ -L １ - and Y is -C(O)-, -S(O) ２ - and -S(O)- are selected, Z １ is selected from O, S and NH, L １ C １～８ Alkylene, C ２～８ Alkenylene and C ２～８ It is alkynylene, L １ Where chemically possible, oxo, halo, C １～３ Alkoxy, C １～３ The method according to claim 24, optionally substituted with one to four substituents selected from haloalkoxys and CNs.

26. The method according to claim 24 or 25, wherein n is 0.

27. The above method involves modified polysaccharide (C): 【Chemistry 11】 A method for forming compound (IV), wherein the compound of formula (IV) is compound (D): 【Chemistry 12】 The method is as follows: Modified polysaccharide (E): 【Chemistry 13】 A method for forming compound (IV), wherein the compound of formula (IV) is compound (F): 【Chemistry 14】 The method according to any one of claims 24 to 26.

28. The method according to any one of claims 24 to 27, wherein the acidic polysaccharide is alginic acid or a pharmaceutically acceptable salt thereof.

29. b) A step of depositing an enzyme substrate of formula (II) onto the modified polysaccharide, wherein formula (II) is as described in any one of claims 12 to 15. The method according to any one of claims 24 to 28, further comprising:

30. A method for forming a wound dressing containing a modified polysaccharide, comprising the steps of forming the modified polysaccharide according to the method of any one of claims 24 to 28, and optionally depositing an enzyme substrate onto the modified polysaccharide according to step b) of the method of claim 29.

31. The method according to claim 30, wherein the acidic polysaccharide forms part of the unmodified wound dressing.

32. A method for forming a packaged wound dressing, i) The step of forming a wound dressing according to the method described in claim 30 or 31, ii) The step of packaging the wound dressing material to form the packaged wound dressing material, iii) Depending on the circumstances, the step of sterilizing the packaged wound dressing and Methods that include...

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