Modified spider silk fibers and their uses

Abstract

Derivatized major ampullate spidroin protein (MaSp)-based fibers are disclosed. Further disclosed are compositions and / or composites comprising the derivatized fibers and methods for making the same.

Description

[Technical Field]

[0001] (Cross-reference of related applications) This application is a U.S. Provisional Patent Application No. 63 / 014,444 filed on April 23, 2020, for the invention titled "Anti-Microbial Compositions"; No. 63 / 048,135 filed on July 5, 2020, for the invention titled "Cosmetic Compositions Comprising Dragline Spider Silk"; No. 63 / 144,089 filed on February 1, 2021, for the invention titled "Cosmetic Compositions Comprising Dragline Spider Silk"; and No. 63 / 144,089 filed on September 17, 2020, for "Modified Spider Silk Fiber and Use The invention claims priority rights under 35 U.S.C. § 119(e) to patent no. 63 / 079,621, entitled “Extruded Compositions Comprising Dragline Spider Silk,” filed on January 6, 2021, and the entirety of their disclosures is incorporated herein by reference.

[0002] In some embodiments, the present invention relates to compositions containing proteins derived from MaSp (large bottle-shaped glandular spidoin) protein, the preparation thereof, and its use as a medicinal cosmetic or antimicrobial material. [Background technology]

[0003] Dragline spider silk is known in the art as the silk used by orbweb-spinning spiders to construct the frame and radius of their webs, as well as to create lifelines for falling or escaping danger. To enable them to perform these tasks, dragline fibers exhibit remarkably high toughness due to a combination of high elasticity and strength, making them one of the toughest fibers, whether natural or artificial. For example, dragline is six times stronger than high-tensile steel in its diameter and three times stronger than Kevlar, one of the strongest synthetic fibers ever created.

[0004] Dragline silk consists of two main polypeptides, usually called large vial-shaped glandular spidoins (MaSp) 1 and 2, and ADF-3 and ADF-4 in the garden spider (Araneus diadematus). These proteins have apparent molecular weights ranging from 200 to 720 kDa, depending on the sample age and analytical conditions. Known dragline silk spidoins consist of highly repeating blocks alternating between alanine-rich segments that form crystalline β-sheets within the fiber and glycine-rich segments that are more flexible and largely lack ordered structure. The C-terminal region is non-repeating and highly conserved across species, forming an α-helix structure. The N-terminal region of dragline silk proteins has also been found to be highly conserved among different spidoins and across different spider species. [Overview of the project]

[0005] According to one embodiment, the derivatized porous large bottle-shaped glandular spidoin protein (MaSp) fiber comprises at least 10 m 2The provided derivative porous MaSp fiber is characterized by a BET surface area of ​​1 / g and comprises a functional moiety covalently bonded to the tyrosine of the porous MaSp fiber, the functional moiety comprising any one of amino, carboxy, nitro, sulfonate, carbonyl, ester, anhydride, carbonate ester, carbamate, cyano, hydroxy, polymer, or any combination thereof.

[0006] In one embodiment, the load of the functionalized portion into the derivatized porous MaSp fiber is 0.01 μmol / g to 10 mmol / g.

[0007] In one embodiment, the functional moiety is covalently bonded to the tyrosine side chain via a diazo bond, a silyl group, or any combination thereof.

[0008] In one embodiment, the polymer is covalently bonded to a chelating agent, an antimicrobial agent, or any combination thereof.

[0009] In one embodiment, the chelating agent comprises (i) a metal chelating group that can bond to a metal or a salt thereof, (ii) a metal oxide chelating group, or both (i) and (ii).

[0010] In one embodiment, the metal chelating group includes iminodiacetic acid (IDA), DOTA, NOTA, NODA, EDTA, HBED-CC (including any salts, derivatives, or combinations thereof).

[0011] In one embodiment, the metal oxide chelating group is selected from salicylic acid, phosphonic acid, hydroxamic acid, malonic acid, pyrogallol, and 5-hydroxy-1,4-naphthoquinone (including any salts or any combination thereof).

[0012] In one embodiment, the polymer is selected from the group consisting of polyglutaraldehyde (PGA), polyvinyl alcohol (PVA), polyacrylic acid (PAA), polyethyleneimine (PEI), polyacrylamide (PAAm), polylysine, polyaniline, polyurethane, polyamide, polyvinyl chloride, silicone crosspolymer, polyvinylpyrrolidone, or any combination thereof.

[0013] In one embodiment, the w / w ratio of the polymer to the porous MaSp fiber is 0.001 to 5.

[0014] In one embodiment, the functional portion is further bonded to a dye or pigment.

[0015] In one embodiment, the MaSp fiber has a decomposition temperature (T) of 280°C to 350°C, as determined by differential scanning calorimetry (DSC). d ), and a glass transition temperature of 200°C to 250°C (T) as determined by DSC. g ) is a characteristic feature.

[0016] In one embodiment, the MaSp fiber is given by formula 10:((X1) Z X2GPGGYGPX3X4X5GPX6GX7GGX8GPGGPGX9X 10 The formula includes a repeating region containing the amino acid sequence shown, where X1 is independently A or G in each case, Z is an integer from 5 to 30, X2 is S or G, X3 is G or E, X4 is G, S or N, X5 is Q or Y, X6 is G or S, X7 is P or R, X8 is Y or Q, X9 is G or S, and X 10 It is either S or G.

[0017] In another embodiment, a composite material is provided comprising the derivatized porous MaSp fibers of the present invention bonded to one or any combination thereof of a metal, a salt thereof, and metal oxide particles.

[0018] In one embodiment, metal, its salt, or metal oxide particles are bonded to a porous MaSp-based fiber that has been derivatized via a chelating agent.

[0019] In one embodiment, the chelating agent comprises (i) a metal chelating group that can bond to a metal or a salt thereof, (ii) a metal oxide chelating group, or both (i) and (ii).

[0020] In one embodiment, the metal chelating group includes iminodiacetic acid (IDA), DOTA, NOTA, NODA, EDTA, HBED-CC (including any salts, derivatives, or combinations thereof), and the metal oxide chelating group is selected from salicylic acid, phosphonic acid, hydroxamic acid, malonic acid, pyrogallol, and 5-hydroxy-1,4-naphthoquinone (including any salts or combinations thereof).

[0021] In one embodiment, the molar ratio of the chelating agent to the derivatized porous MaSp fiber is 0.01 to 1.

[0022] In one embodiment, the metal oxide particles are selected from titania, zirconia, silica, or any combination thereof. In one embodiment, the metal oxide particles are characterized by a particle size of 10 to 5,000 nm.

[0023] In one embodiment, the w / w ratio of derivatized porous MaSp fibers to metal oxide particles in the composite material is 0.01 to 100.

[0024] In another embodiment, a method for synthesizing a derivatized porous MaSp fiber of the present invention, wherein the porous MaSp fiber is given formula I: [ka] Or formula II: [ka] A method is provided for obtaining a derivatized porous MaSp fiber, comprising the step of reacting with a reagent represented by the formula, where R comprises the functional part of the present invention, A is selected from substituted or unsubstituted aryl, heteroaryl, and alkyl (including any combination thereof), each R1 independently comprises one of hydrogen, alkyl, hydroxy, or alkoxy (including any combination thereof), the wavy bond represents a linker or bond, and the reacting step comprises conditions sufficient to covalently bond the functional part to the tyrosine of a porous MaSp fiber.

[0025] In one embodiment, the covalent bond is mediated by a diazo bond or C-Si.

[0026] In one embodiment, R1 is -OC 1~10 Contains alkyl.

[0027] In one embodiment, A includes substituted or unsubstituted phenyl.

[0028] In one embodiment, the linker is substituted or unsubstituted C 1~10 Contains alkyl.

[0029] In one embodiment, the method further includes a step of reacting a functional portion with a polymer that is reactive to the functional portion, thereby covalently bonding the polymer to the porous MaSp fiber, wherein the functional portion contains an amine or a carboxyl group.

[0030] In one embodiment, the method further includes the step of reacting the polymer with a chelating agent that is reactive with the polymer, thereby obtaining the chelating agent covalently bonded to the polymer.

[0031] In one embodiment, the polymer includes PGA.

[0032] In another embodiment, a composition is provided comprising a large bottle-shaped glandular spidoin protein (MaSp) system fiber containing a microbial interaction protein.

[0033] In some embodiments, the microbial interaction protein is a viral interaction protein. In some embodiments, the viral interaction protein is a viral-binding receptor. In some embodiments, the virus is a coronavirus.

[0034] In some embodiments, the composition further comprises at least one antimicrobial agent. In some embodiments, the antimicrobial agent is selected from the group consisting of reactive oxygen species (ROS) sources, carboxylic acids, quaternary amines, bactericides, transition metals, electrophilic reactive groups, oxidizing agents, antimicrobial polymers, or any combination thereof.

[0035] In some embodiments, the antimicrobial agent is bonded to the MaSp fiber by hydrogen bonds, van der Waals bonds, or both. In some embodiments, the MaSp fiber is coated with a polymer layer, which is further bonded to a transition metal. In some embodiments, the antimicrobial agent is covalently bonded to the MaSp fiber. In some embodiments, the covalent bond is mediated through a linker containing a diazo group. In some embodiments, the MaSp fiber is covalently bonded to a polymer containing a chelate moiety. In some embodiments, the polymer contains polyglutaraldehyde, and the chelate moiety contains iminodiacetic acid.

[0036] In some embodiments, the composition includes a transition metal bonded to a chelate portion. In some embodiments, the transition metal is in further contact with a further transition metal.

[0037] In another embodiment, a composition is provided comprising a large vial-shaped glandular spidoin protein (MaSp) system fiber modified with any one of an antimicrobial agent and a transition metal. In some embodiments, the antimicrobial agent is selected from the group consisting of reactive oxygen species (ROS) sources, carboxylic acids, quaternary amines, bactericides, transition metals, electrophilic reactive groups, oxidizing agents, antimicrobial polymers, or any combination thereof.

[0038] In some embodiments, the modification includes covalent, non-covalent, or both.

[0039] In some embodiments, the MaSp fibers are coated with a polymer layer, which is further bonded to a transition metal. In some embodiments, the covalent bond is mediated through a linker containing a diazo group. In some embodiments, the MaSp fibers are covalently bonded to a polymer containing a chelate moiety.

[0040] In some embodiments, the composition includes a transition metal bonded to the chelate portion. In some embodiments, the transition metal is in further contact with a further transition metal. In some embodiments, the polymer includes polyglutaraldehyde, and the chelate portion includes iminodiacetic acid.

[0041] In some embodiments, the compositions or MaSp fibers of the present invention are characterized by porosity suitable for capturing microorganisms in the range of 50 to 500 nm. In some embodiments, the porosity includes median pore diameters in the range of 1 to 500 nanometers.

[0042] In some embodiments, the ratio of MaSp fiber to antibacterial agent is 0.01:1 to 1:1.

[0043] In some embodiments, the composition is intended for use in providing an antimicrobial effect to one of a cream, fabric, or substrate.

[0044] In some embodiments, the composition comprises 0.01% to 50% (w / w) of MaSp fibers and further polymers.

[0045] In some embodiments, further polymers are selected from the group consisting of thermoplastic polymers, thermosetting polymers, epoxy, polyester, polyamide, polyol, polyurethane, polyethylene, nylon, polyacrylate, polycarbonate, polylactic acid (PLA) or its copolymers, silicone, liquid crystal polymer, maleic anhydride grafted polypropylene, polycaprolactone (PCL), rubber, cellulose, or any combination thereof.

[0046] In another embodiment, an article comprising the composition described herein is provided. In some embodiments, the article further comprises 0.01% to 50% w / w of additional polymers.

[0047] In some embodiments, the article is in the form of a fibrous material, a polymer layer, or a coating. In some embodiments, the article is characterized by at least one improved antimicrobial property compared to an article without the composition.

[0048] Unless otherwise specified, all technical and / or scientific terms used herein have the same meaning as commonly understood by those skilled in the art in the industry to which the present invention pertains. Similar or equivalent methods and materials may be used in the implementation or testing of embodiments of the present invention, but exemplary methods and / or materials are described below. In case of any conflict, the present specification, including definitions, shall prevail. Furthermore, materials, methods, and examples are illustrative and not necessarily limiting.

[0049] Further embodiments and the full scope of the present invention will become apparent from the detailed description given below. However, it should be understood that the detailed description and specific examples illustrating preferred embodiments of the present invention are given merely as examples, as various variations and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. [Brief explanation of the drawing]

[0050] [Figure 1] This describes the multilayer structure and stepwise preparation process of Cu-coated MsSp fibers (referred to herein as SVXE). [Figure 2] This image shows a nylon fiber reinforced with SVXE coated with Cu (10% w / w). [Figure 3] Figures 3A and 3B show micrographs of metal-coated SVXE fibers. Figure 3A: Micrograph of Pd-doped SVXE fiber. Figure 3B: Micrograph showing a Cu layer (Cu-SVXE) deposited on a Pd-doped SVXE fiber. [Figure 4] This image shows the antimicrobial properties of nylon fibers reinforced with 10% w / w Cu-SVXE compared to nylon fibers in their original state. ZOI (Zone of Inhibition) indicates areas where bacterial growth is absent. [Figure 5] This graph shows the IR spectra of carboxylated MaSp fibers (derivativeized via aminobenzoic acid through a diazo bond) and the original MaSp fibers. * indicates peaks corresponding to the OH stretching vibration of the carboxyl group. The IR spectrum of carboxylated MaSp fibers shows increased intensity of the corresponding peaks compared to the original MaSp fibers, supporting efficient carboxylation of MaSp fibers. ** indicates peaks corresponding to the CH stretching vibration of aromatic hydrogens in the tyrosine side chains. The IR spectrum of carboxylated MaSp fibers shows a significant decrease in peak intensity compared to the original MaSp fibers, supporting efficient tyrosine diazotization of carboxylated MaSp fibers. [Figure 6] This graph shows the zeta potentials of amination-modified MaSp fibers (amine), carboxylated MaSp fibers (acid), PAA-modified amination-modified MaSp fibers (PAA), PEI-modified carboxylated MaSp fibers (PEI), and the original MaSp fibers (SVXE). [Figure 7] Figures 7A and 7B are SEM images of porous MaSp fibers bonded to polyacrylate (Figure 7A) or polyglutaraldehyde (Figure 7B). [Figure 8] A graph showing the differential scanning calorimetry (DSC) curves of SVXE at a temperature increase from 25°C to 280°C (curve 1), cooling to 50°C (curve 2), and a new increase to 350°C (curve 3).

Mode for Carrying Out the Invention

[0051] In some embodiments, the present invention relates to chemically modified or derivatized MaSp-based fibers. As demonstrated below in this specification, the inventors have synthesized conjugates of derivatized MaSp-based fibers (e.g., containing functional moieties attached to tyrosine of MaSp-based fibers via diazo bonds or silyl groups as described below in this specification) with various polymers such as PGA, PAA, PVA, PEI, PAAm, or combinations thereof (such as PGA-co-PEI). Exemplary and non-limiting derivatized MaSp-based fibers such as MaSp-based fibers modified with an amino or PEI having a cationic group exhibited a positive zeta potential. Surprisingly, MaSp-based fibers characterized by a positive zeta potential showed excellent adhesion to hydrophobic surfaces such as hair (e.g., damaged hair) and glass surfaces.

[0052] Furthermore, the inventors utilized some of the derivatized MaSp-based fibers of the present invention to complex a metal (such as the Cu element) in order to obtain MaSp-based fibers characterized by excellent conductivity.

[0053] According to one aspect, the present invention provides a derivatized porous major ampullate spidroin (MaSp)-based fiber, comprising a derivatized porous MaSp-based fiber containing a functional moiety covalently bonded to an amino acid of the porous MaSp-based fiber. In some embodiments, the porous MaSp-based fiber is at least 10 m 2It is characterized by a BET surface area of ​​ / g. In some embodiments, the porous MaSp fiber is as described below in this specification. In some embodiments, the derivatized porous MaSp fiber includes a functional moiety covalently bonded to the amino acid side chain of the porous MaSp fiber, and the porous MaSp fiber is at least 10m 2 It is characterized by a BET surface area of ​​ / g.

[0054] In some embodiments, the derivatized porous MaSp fiber comprises a functional moiety covalently bonded to the amino acid side chain of the porous MaSp fiber, and the porous MaSp fiber comprises (i) at least 10 m 2 The material is characterized by a BET surface area of ​​1 / g, (ii) a decomposition temperature of 280-350°C, (iii) a glass transition temperature of 200-250°C, and (iv) at least one of the amino acid sequences in which each repeat region independently comprises an amino acid sequence shown in the following formula 10, or a combination of (i), (ii), (iii), and (iv).

[0055] In some embodiments, the derivatized porous MaSp fiber comprises a functional moiety covalently bonded to the amino acid side chains of the porous MaSp fiber, the porous MaSp fiber comprises repeating regions, each repeating region independently comprises the amino acid sequence shown in the following formula 10, and the derivatized porous MaSp fiber comprises (i) at least 10 m 2 The fiber is characterized by a BET surface area of ​​1 / g, (ii) a decomposition temperature of 280–350°C, and (iii) a glass transition temperature of 200–250°C, or a combination of (i), (ii), and (iii). In some embodiments, the terms “porous MaSp fiber” and “MaSp fiber” are used interchangeably herein.

[0056] In some embodiments, the functional moiety is covalently bonded to an amino acid of a porous MaSp fiber. In some embodiments, the functional moiety is covalently bonded to an amino acid, which is selected from tyrosine, serine, cysteine ​​and lysine, threonine, histidine, arginine, aspartic acid and glutamic acid, or any combination thereof. In some embodiments, the functional moiety is covalently bonded to the side chain of an amino acid, which is selected from tyrosine, serine, cysteine ​​and lysine, threonine, histidine, arginine, aspartic acid and glutamic acid, or any combination thereof. In some embodiments, the functional moiety is covalently bonded to the side chain of tyrosine, serine, cysteine ​​and lysine, or any combination thereof. In some embodiments, the functional moiety is covalently bonded to the side chain of tyrosine (e.g., a phenol ring).

[0057] In some embodiments, the derivatized porous MaSp fibers of the present invention include a functional moiety covalently bonded to at least one tyrosine side chain via a diazo bond, a silyl group, or both. In some embodiments, the derivatized porous MaSp fibers of the present invention include diazotized tyrosine, silylated tyrosine, or both. In some embodiments, 1-99%, 10-99%, 10-90%, 10-80%, 10-70%, and 10-60% (including any range or value in between) of tyrosine residues (i.e., side chains) in the derivatized porous MaSp fibers of the present invention are diazotized and / or silylated.

[0058] In some embodiments, the derivatized porous MaSp fibers of the present invention include a functional moiety (also referred to herein as "diazotized tyrosine") covalently bonded to the side chain of at least one tyrosine via a diazo bond. In some embodiments, the diazotized tyrosine is bonded via a diazo bond: [ka] It contains a functional moiety covalently bonded to the phenyl ring of tyrosine via [a specific linkage].

[0059] In some embodiments, the functionalized portion is covalently bonded to the porous MaSp fiber via one of the following: diazo, silyl, carbonyl, amide, ester, maleimide, or any combination thereof.

[0060] In some embodiments, the functional moiety is covalently bonded to at least one tyrosine residue of a porous MaSp fiber. In some embodiments, the functional moiety is covalently bonded to at least one tyrosine residue via a bond selected from diazo, silyl, ester, carbamate, carbonyl, (O or S)-thiocarbamate, or any combination thereof. In some embodiments, the functional moiety is covalently bonded to at least one tyrosine residue via a diazo bond. In some embodiments, the functional moiety is covalently bonded to at least one tyrosine residue via a linker containing any reactive group that can form a diazo bond (e.g., an alkyldiazonium or aryldiazonium group) with the phenol ring of tyrosine, and / or (ii) a Si-C bond (e.g., an alkoxysilyl or halosilyl) with the phenol ring of tyrosine.

[0061] In some embodiments, the composition substantially lacks functionalized moieties and / or polymers adsorbed onto MaSp fibers.

[0062] In some embodiments, the derivatized porous MaSp fiber of the present invention, which includes a functionalized moiety covalently bonded to the MaSp fiber via a diazo bond, is given by formula 1: [ka] It is expressed by, in the formula, [ka] ∫ represents a MaSp fiber, where A is selected from aryl (e.g., bicyclic / condensed aromatic rings such as phenyl or naphthalene), heteroaryl (e.g., C5-6 aromatic rings containing 1, 2, 3 or 4 heteroatoms selected from O, N and S), and substituted or unsubstituted alkyl (e.g., C1-C10 linear or branched alkyl), where R is the functional part of the present invention or contains it, and wavy bond represents a linker (or spacer) as described herein.

[0063] In some embodiments, the derivatized porous MaSp fiber of the present invention, which includes a functional moiety covalently bonded to the MaSp fiber via a silyl group, is given by formula 2: [ka] Represented by the formula, where the wavy bond and R are as described herein, and each R1 independently comprises one of the following: hydrogen, optionally substituted alkyl (e.g., C1-C10 linear or branched alkyl), hydroxyl, or alkoxy (e.g., C1-C10 alkoxy).

[0064] In some embodiments, the functional moiety of the present invention comprises one of the following: a hydroxyl group, a mercapto group, an amino group, a carboxylate group, a nitro group, a sulfonate group, a carbonyl group, an anhydride, a carbonate ester, a carbamate group, a polymer, or any combination thereof. In some embodiments, the functional moiety comprises a hydroxyl group, a mercapto group, an amino group, a carboxylate group, and a polymer, or any combination thereof. In some embodiments, the functional moiety is covalently bonded to the amino acids of the porous MaSp fiber via a linear or branched linker. In some embodiments, each derivatized amino acid of the porous MaSp fiber comprises one or more functional moieties covalently bonded thereto.

[0065] In some embodiments, the functionalized portion of the present invention is linked to a tyrosine residue via a diazoaryl linker. In some embodiments, the functionalized portion of the present invention is linked to a diazoaryl linker. In some embodiments, the terms “linker” and “spacer” are used interchangeably herein.

[0066] In some embodiments, the functionalized moiety (or R) comprises electrophilic and / or nucleophilic groups. Numerous electrophiles and nucleophiles are well known in the art.

[0067] Non-limiting examples of electrophilic reactive groups include, but are not limited to, aldehydes, ketones, carboxyls, esters, imines, oximes, acyl halides, active esters (e.g., N-hydroxysuccinimide), chloroformates, anhydrides, epoxides, isocyanates, nitros, sulfonates, trialkylammoniums, and any alkyl and / or aryl derivatives (such as carboxyalkyls, carboxyaryls, alkylcarbonyls, haloalkyls, haloaryls, etc.) or any combination thereof.

[0068] Non-exclusive examples of nucleophilic reactive groups include, but are not limited to, hydroxy, mercapto, amino, phosphine, or any combination thereof.

[0069] In some embodiments, the functionalized moiety comprises multiple electrophilic and / or nucleophilic groups. In some embodiments, the functionalized moiety comprises a polyamine (e.g., a linear polyamine such as spermine or spermidine, or a branched polyamine such as tris(2-aminoethyl)amine). In some embodiments, the functionalized moiety comprises a polyol (e.g., pentaerythritol, xylitol). In some embodiments, the functionalized moiety comprises a di or tricarboxylic acid (e.g., citric acid, malic acid, succinic acid).

[0070] In some embodiments, the functional moiety (or R) is selected from alkyl groups (e.g., linear or branched C1-10 alkyl groups such as methyl, ethyl, propyl, butyl, isobutyl, pentyl, isopentyl, hexyl, heptyl, octyl, nonyl, decyl), C1-10 alkoxy groups (e.g., methoxy, ethoxy, propoxy, octyloxy), vinyl groups (e.g., vinyloxy), C1-10 alkylamino groups (e.g., methylamino, ethylamino, propylamino, butylamino, pentylamino, hexylamino), and protected amines (e.g., Fmoc, Boc, benzyl, CBz, etc.). It comprises any one of the following: amine protecting group, hydroxy (optionally protected by a hydroxy protecting group such as tert-butyl or trimethylsilyl), mercapto (optionally protected by a thiol protecting group), silyl, siloxane (e.g., trialkoxysilane), nitro, sulfonate, cyano, halo, trialkylammonium, aldehyde, ketone, carboxy, ester, imine, oxime, halide acyl, active ester (e.g., N-hydroxysuccinimide), chloroformate, anhydride, epoxide, isocyanate, or any combination thereof.

[0071] In some embodiments, the linker (or spacer) comprises an alkyl optionally substituted with any one of the following: carboxy, halo, hydroxy, amino, cycloalkyl, alkyl, nitro, sulfonate, cyano, or any combination or derivative thereof. In some embodiments, the linker comprises an optionally substituted alkoxy, thioalkyl, aminoalkyl, glycol, or any combination thereof. In some embodiments, alkyls comprising any derivative thereof are as described below herein. In some embodiments, the linker comprises a disubstituted alkyl (e.g., aminohexanoic acid) or a disubstituted heteroalkyl group. In some embodiments, the linker comprises C1-C10 alkyl, C1-C10 aminoalkyl, C1-C10 alkoxy, C1-C10 mercaptoalkyl, and carbonyl derivatives (e.g., -C(O)NH-, -C(O)O-, -C(O)-, -C(O)S-, -C(NH)NH-, -C(NH)O-, -C(NH)S-), and optionally contains one or more heteroatoms (e.g., S, N, O) within the framework of the linker or any combination thereof.

[0072] In some embodiments, the linker (or spacer) comprises natural and / or unnatural amino acids, alkyl, amide bonds, ester bonds, thioester bonds, and urea bonds (including any derivatives or combinations thereof). In some embodiments, the linker of the present invention comprises a click reaction product (e.g., a succinimide-thioether moiety formed via covalent bonding and / or a click reaction, such as a cyclization reaction product). Further linkers or spacers are well known in the art.

[0073] Click reactions are well known in the art and include, in particular, Michael addition of maleimides and thiols (resulting in the formation of succinimide-thioethers), azido-alkynecycloaddition, Diels-Alder reactions (e.g., direct and / or inverse electron-requiring Diels-Alder), dibenzylcyclooctin 1,3-nitrone (or azido)cycloaddition, and alkene-tetrazole photoclick reactions.

[0074] As used herein, the term “heteroalkyl” refers to an alkyl group as defined herein, in which one or more of its constituent carbon atoms are substituted with nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkyl group may be further substituted with one, two, three, or four substituents, as described herein for alkyl groups. Examples of heteroalkyl groups are polyglycols or polyalkoxylates, such as polyethylene glycol.

[0075] In some embodiments, the derivatized porous MaSp-based fibers of the present invention are as follows: [ka] As shown in the formula, where the wavy bond represents a linker, and the linker is as described herein. In some embodiments, R is as described herein. In some embodiments, the linker is or contains an optionally substituted C1-C6 or C1-C10 alkyl group. In some embodiments, R includes amino, halo, nitro, carbonyl, ester, or carboxyl groups.

[0076] In some embodiments, the derivatized porous MaSp-based fibers of the present invention are as follows: [ka] As shown in the formula, where R is as described herein, n, m and o are each independently integers from 0 to 20, and each R2 is independently H, or halogen, -NO2, -CN, -OH, -CONH2, -CONR2, -CNNR2, -CSNR2, -CONH-OH, -CONH-NH2, -NHCOR, -NHCSR, -NHCNR, -NC(=O)OR, -NC(=O)NR, -NC(=S)OR, -NC(=S)NR, -SO2R, -SOR, -SR, -SO2OR, -SO2N(R)2, -NHNR2, -NNR, C1-C6 haloalkyl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1-C6 alkyl) -N(C1~C6alkyl)2, C1~C6alkoxy, C1~C6haloalkoxy, hydroxy(C1~C6alkyl), hydroxy(C1~C6alkoxy), alkoxy(C1~C6alkyl), alkoxy(C1~C6alkoxy), C1~C6alkyl-NR2, C1~C6alkyl-SR, -CONH(C1~C6alkyl), -CON(C1~C6alkyl)2, -CO2H, -CO2R, -OCOR, -OCOR, -OC(=O)OR, -OC(=O)NR, -OC(=S)OR, -OC(=S)NR or a combination thereof, and X1 is a heteroatom (e.g., O, S, N, NH) or absent. In some embodiments, n, m, and o are each independently integers of 0 or 0-20, 1-20, 1-3, 3-5, 5-7, 7-10, 10-15, 15-20 (including any range between them). In some embodiments, R is amino, carboxy, [ka] or a combination thereof, including the formula where X1 is as described herein, R3 is H, and C1-C are optionally substituted. 10 Alkyl, optionally substituted aryl, optionally substituted heteroaryl, or any combination thereof, or containing thereof. In some embodiments, X1 is N or NH, or containing thereof. In some embodiments, R2 is H or optionally substituted C1-C6 alkyl, or containing thereof.

[0077] In some embodiments, the derivatized porous MaSp-based fibers of the present invention are as follows: [ka] As shown in the formula, n, R2 and R3 are as described herein.

[0078] In some embodiments, the derivatized porous MaSp-based fibers of the present invention are as follows: [ka] As shown in the formula, where R2 and n are as described above in this specification.

[0079] In some embodiments, the derivatized porous MaSp-based fibers of the present invention are as follows: [ka] As shown in the formula, where R and n are as described herein, and each R1 is independently hydrogen, alkyl (e.g., C1-C6 alkyl or C1-C 10 Alkyl, hydroxy, or C1-C 10 It contains one of the following alkoxys (e.g., methoxy, ethoxy, propoxy, pentoxy, etc.).

[0080] In some embodiments, the derivatized porous MaSp-based fibers of the present invention are as follows: [ka] As shown in the formula, R1, n, R3, and R2 are as described herein.

[0081] In some embodiments, the functional moiety is covalently bonded to one of histidine, arginine, aspartic acid, glutamine, or any combination thereof. Those skilled in the art will understand that the functional moiety can be covalently bonded to a carboxylic acid side chain residue via an amino, hydroxy, or mercapto-substituted linker.

[0082] In some embodiments, the derivatized porous MaSp fibers are as follows: [ka] As shown in the formula, where X represents a side-chain heteroatom selected from O, S, and N, and R is as described above herein. In some embodiments, X represents a heteroatom of any one of the amino acids of the MaSp fiber.

[0083] In some embodiments, the derivatized porous MaSp fibers are as follows: [ka] As shown in the formula, where X represents one of the side chain heteroatoms of the amino acids in the MaSp fiber, [ka] represents a bonding point to the MaSp fiber, the wavy bond represents a linker, and R is as described herein. In some embodiments, R includes amino, halo, nitro, carbonyl, ester, or carboxy.

[0084] In some embodiments, the functional portion is covalently bonded to the hydroxyl group of at least one tyrosine residue of the MaSp fiber.

[0085] In some embodiments, the derivatized porous MaSp fibers are as follows: [ka] As shown in the formula, where X is as described herein.

[0086] In some embodiments, the functional moiety is covalently bonded to at least one amino acid of the MaSp fiber via a silyl bond. In some embodiments, the amino acid is a nucleophilic amino acid (e.g., serine, cysteine, threonine, and lysine). In some embodiments, the derivatized porous MaSp fiber is as follows: [ka] As shown in the formula, where X represents one of the side-chain heteroatoms (e.g., S, NH, or O) of the amino acids in the MaSp fiber, [ka] represents a bonding point to the MaSp fiber, the wavy bond represents a linker, R is as described herein, and each R1 independently contains one of hydrogen, alkyl, hydroxy, or alkoxy.

[0087] In some embodiments, the derivatized porous MaSp fibers are as follows: [ka] As shown in the formula, where X represents one of the side chain heteroatoms of the amino acids in the MaSp fiber, [ka] represents a binding site to the MaSp fiber. In some embodiments, X represents a cysteine ​​and / or tyrosine side chain heteroatom.

[0088] In some embodiments, the derivatized porous MaSp fibers are as follows: [ka] As shown in the formula, where X represents a heteroatom selected from O, S, and N, and R is as described above in this specification.

[0089] In some embodiments, the functional moiety is covalently bonded to the MaSp fiber via at least one nucleophilic amino acid selected from serine, cysteine, threonine, and lysine. In some embodiments, the functional moiety is covalently bonded to the nucleophilic amino acid of the MaSp fiber via a bond selected from silyl, ester, carbamate, carbonyl, (O or S)-thiocarbamate, or a combination thereof.

[0090] In some embodiments, the functionalized moiety imparts reactivity to the MaSp fiber. Those skilled in the art will understand that the functionalized moiety, such as an amine, can react with electrophiles (e.g., haloalkyl or ester). Therefore, by introducing the functionalized moiety into the MaSp fiber, the MaSp fiber can subsequently react with any reagent (e.g., small molecule or polymer) that is reactive to the functionalized moiety. In some embodiments, the functionalized moiety induces or increases the reactivity of the MaSp fiber. In some embodiments, the functionalized moiety induces or increases the reactivity of the MaSp fiber to any reagent that can react with it.

[0091] In some embodiments, the derivatized porous MaSp fibers of the present invention include a functional moiety (e.g., amino or carboxy) covalently bonded to a polymer. In some embodiments, the polymer is covalently bonded to the functional moiety via a functional group that is reactive to the functional moiety (e.g., the functional moiety is carboxy or contains carboxy and the functional group of the polymer is amino, mercapto or hydroxy, or the functional moiety is amino or contains amino and the functional group of the polymer is halo, carbonyl or carboxy). Those skilled in the art will understand that there are further known reactive groups (e.g., via click reactions) that can be used for the covalent bonding of the polymer to the functional moiety of the present invention. In some embodiments, the polymer is covalently bonded to the functional moiety via any of -C(O)NH-, -C(O)O-, -C(O)-, -C(O)S-, -C(NH)NH-, -C(NH)O-, -C(NH)S-, -NC(O)-, -N(C)- or a combination thereof.

[0092] In some embodiments, the functionalized portion of the derivatized porous MaSp fiber is covalently bonded to the polymer. In some embodiments, the polymer is positively and / or negatively charged. In some embodiments, the polymer is neutral.

[0093] In some embodiments, the polymer is selected from cationic polymers (e.g., PEI, polylysine, polyarginine, chitosan) (including any derivatives and / or copolymers thereof), anionic polymers (e.g., PAA), and / or nonionic polymers (e.g., PVA, PVC, silane, polyamide). Other cationic polymers, anionic polymers, and / or nonionic polymers are well known in the art.

[0094] In some embodiments, the polymer is selected from the group consisting of polyglutaraldehyde (PGA), polyvinyl alcohol (PVA), polyacrylic acid (PAA), polyacrylate, linear or branched polyethyleneimine (PEI), polyacrylamide (PAAm), polylysine, polyarginine, polyaniline, polyurethane, polyamide (e.g., nylon), polyvinyl chloride, polysilane, chitosan, N-haramine polymer, N-haramid polymer, polysilane-co-polyolefin, silane-crosslinked polyolefin, and polyvinylpyrrolidone (PVP) (including any combination or copolymer thereof). In some embodiments, the polymer is a linear polymer. In some embodiments, the polymer is a branched polymer. In some embodiments, the polymer is a copolymer. In some embodiments, the polymer is a graft copolymer.

[0095] In some embodiments, the polymer is covalently bonded to a nucleophilic or electrophilic functional group of a derivatized porous MaSp fiber. Those skilled in the art will understand that derivatized MaSp fibers can be obtained by reacting aminated MaSp fibers with a carboxyl group of a polymer (e.g., PAA) or a carbonyl group of a polymer (e.g., PGA) to obtain derivatized MaSp fibers covalently bonded to a polymer. Furthermore, carboxylated MaSp fibers can be reacted with an amino group of a polymer (e.g., PEI) or a hydroxyl group of a polymer (e.g., PVA). The inventors have succeeded in synthesizing conjugates of derivatized MaSp fibers (including, for example, functional moieties bonded to tyrosine of the MaSp fiber via diazo bonds or silyl groups, as described above) with various polymers such as PGA, PAA, PVA, PEI, PAAm, or combinations thereof (e.g., PGA-co-PEI). Furthermore, the inventors have succeeded in synthesizing the above conjugates using MaSp proteins having mutated amino acid sequences (also used herein as "mutated MaSp proteins").

[0096] The inventors of this invention have proposed formula 3: [ka] We successfully synthesized PAAm-modified derivatized MaSp fibers by in-situ polymerization on amination-modified MaSp fibers represented by [formula]. In some embodiments, the PAAm-modified derivatized MaSp fibers are as follows: [ka] As shown in the equation, each n is independently 0-10, 0-20, 1-20, 1-3, 3-5, 5-7, 7-10, 10-15, 15-20 (including any range between them). In some embodiments, m is 1-10000, 1-10, 10-100, 100-1000, 1000-10000 (including any range between them).

[0097] In some embodiments, the w / w ratio of MaSp fibers to the polymer is 0.01:1 to 1:1, 0.02:1 to 1:1, 0.05:1 to 1:1, 0.09:1 to 1:1, 0.1:1 to 1:1, 0.5:1 to 1:1, or 0.9:1 to 1:1 (including any range in between).

[0098] In some embodiments, the w / w ratio of MaSp fibers to the polymer is 100:1~1:100, 95:1~1:100, 80:1~1:100, 60:1~1:100, 50:1~1:100, 30:1~1:100, 20:1~1:100, 10:1~1:100, 9:1~1:100, 5:1~1:100, 2:1~1:100, 100:1~1:80, 95:1~1:80, 80:1~1:80, 60:1~1:80, 50:1~1:80, 30:1~1:80, 20:1~1:80, 10:1~1:80, 9:1~1:80, 5:1~1 :80, 2:1~1:80, 100:1~1:50, 95:1~1:50, 80:1~1:50, 60:1~1:50, 50:1~1:50, 30:1~1:50, 20:1~1:50, 10:1~1:50, 9:1~1:50, 5:1~1:50, 2:1~1:50, 100:1~1:10, 95:1~1:10, 80:1~1:10, 60:1~1:10, 50:1~1:10, 30:1~1:10, 20:1~1:10, 10:1~1:10, 9:1~1:10, 5:1~1:10, or 2:1~1:10 (including any range in between).

[0099] In some embodiments, the functional groups of the derivatized MaSp fibers are bonded to a polymer containing multiple reactive groups. In some embodiments, the reactive groups include nucleophilic groups (such as amino, hydroxy, and thiol), electrophilic groups (such as carbonyl, carboxy, ester, succinimide ester, halo, nitro, and azide), or both. In some embodiments, the polyaldehyde polymer (e.g., polyglutaraldehyde) bonded to the MaSp fibers is as follows: [ka] As shown in the equation, dashed lines represent any combination, and m and n are integers. In some embodiments, each n is independently 0-10, 0-20, 1-20, 1-3, 3-5, 5-7, 7-10, 10-15, 15-20 (including any range between them). In some embodiments, m is 1-10000, 1-10, 10-100, 100-1000, 1000-10000 (including any range between them).

[0100] In some embodiments, the derivatized MaSp fibers are bonded to a polymer containing multiple chelating agents. In some embodiments, the chelating agents include (i) a metal chelating group that can bond to a metal or a salt thereof, (ii) a metal oxide chelating group, or both (i) and (ii).

[0101] In some embodiments, a metal chelating group can complexate a metal or a salt thereof (by coordinate bonding).

[0102] In some embodiments, the metal or its salts include transition metals. Non-limiting examples of transition metals include, but are not limited to, gold (Au), copper (Cu), palladium (Pd), zinc (Zn), aluminum (Al), tungsten (W), titanium (Ti), silicon (Si), zirconium (Zr), hafnium (Hf), tin (Sn), gallium (Ga), molybdenum (Mo), nickel (Ni), vanadium (V), platinum (Pt), tantalum (Ta), germanium (Ge), and niobium (Nb), or any combination thereof.

[0103] In some embodiments, the metal chelating group includes thiols, amines, phenols, and carboxyls (including any derivatives thereof). In some embodiments, the metal chelating group includes crown ethers. In some embodiments, the metal chelating group is a cyclic molecule comprising multiple carboxyl and / or hydroxyl groups configured to complex a metal or a salt thereof. In some embodiments, metal chelating groups including DOTA, NOTA, NODA, EDTA, and HBED-CC (including any salts, derivatives, or combinations thereof) are well known in the art. In some embodiments, the metal chelating group includes iminodiacetic acid (IDA), its salts, or derivatives.

[0104] In some embodiments, polymers containing multiple metal chelate groups include: [ka] This is expressed as follows, where CM represents the chelating agent, k is 10-10000, 10-100, 100-1000, 100-10000 (including any range in between), m is 0-10, 0-3, 3-5, 5-10 (including any range in between), and each n independently represents an integer between 0-10, 0-20, 1-20, 1-3, 3-5, 5-7, 7-10, 10-15, 15-20 (including any range in between).

[0105] In another embodiment, the derivatized MaSp fiber comprises a functional moiety (e.g., a polymer) bonded to a metal oxide chelate group, wherein the functional moiety is as described herein. In some embodiments, the metal oxide chelate group has an affinity for metal oxides or particles containing them. In some embodiments, the metal oxide chelate group can form a complex with metal oxides or particles containing them (e.g., by coordination bonds). In some embodiments, the metal oxide particles are as described herein.

[0106] In some embodiments, the term “complexing” refers to the stable (e.g., chemically stable) complexation of a metal and / or metal oxide (including any particles containing it). In some embodiments, stable complexation refers to the ability of a metal-bonded derivatized MaSp fiber (also referred to herein as “composite” or “stable composite”) to retain at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% of its initial metal content upon exposure to a solvent (e.g., organic solvent, aqueous solvent, etc.) or storage under ambient conditions for a period of at least 1 month (m), at least 2 m, at least 6 m, at least 12 m, at least 2 years (y), at least 3 y, at least 10 y (including any range in between).

[0107] In some embodiments, the weight / weight (w / w) ratio of the functional portion in the derivatized fiber of the present invention to the porous MaSp fiber is 0.01-30%, 0.01-0.1%, 0.1-0.5%, 0.5-1%, 1-5%, 5-10%, 10-20%, 20-30% (including any range in between).

[0108] In some embodiments, the loading of the functionalized portion into the derivatized porous MaSp fiber is 0.01 μmol / g to 10 mmol / g, 0.01 μmol / g to 0.1 μmol / g, 0.1 μmol / g to 0.5 μmol / g, 0.5 μmol / g to 1 μmol / g, 1 μmol / g to 10 μmol / g, 10 μmol / g to 30 μmol / g, 30 μmol / g to 50 μmol / g, 50 μmol / g to 100 μmol / g, 100 μmol / g to 500 μmol / g, 0.5 to 1 mmol / g, 1 to 5 mmol / g, and 5 to 10 mmol / g (including any range in between).

[0109] In some embodiments, at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 80%, at least 90%, at least 95%, and at least 99% (including any range in between) of the tyrosine residues in the derivatized MaSp fiber of the present invention are replaced by functional moieties, where the functional moieties are as described herein. In some embodiments, the degree of tyrosine residue substitution in the derivatized MaSp fiber is 1-99%, 1-5%, 5-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, and 90-99% (including any range in between).

[0110] In some embodiments, the derivatized porous MaSp fiber of the present invention includes a functional moiety covalently bonded to at least one tyrosine side chain such that the degree of substitution of tyrosine residues (i.e., side chains) within the derivatized MaSp system is 1-90%, 1-99%, 10-99%, 10-90%, 10-80%, 10-70%, 10-60% (including any range or value in between).

[0111] In some embodiments, the degree of substitution of tyrosine residues (i.e., side chains) within the derivatized MaSp fibers is up to 90%, up to 80%, up to 70%, up to 65%, and up to 60% (including any range in between). The inventors have succeeded in substituting up to approximately 60% of tyrosine residues (i.e., side chains) with various functional moieties (some of which are described in the Examples section).

[0112] In some embodiments, the functionalized moiety is selectively bound to at least one tyrosine residue of the MaSp fiber. In some embodiments, the selectivity includes at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, and at least 99% (including any range in between).

[0113] In some embodiments, the functionalized portion imparts a positive and / or negative charge to the MaSp fiber. In some embodiments, the functionalized portion modifies the surface charge of the MaSp fiber. In some embodiments, the functionalized portion modifies the properties of the MaSp fiber, which are selected from wettability, water contact angle, dispersibility, or solubility (e.g., in water and / or organic solvents).

[0114] In some embodiments, derivatized (e.g., amino-derivatized) porous MaSp fibers are characterized by a positive zeta potential of 1 to 50 at a pH of about 7. As illustrated herein, amination-derivatized MaSp fibers are characterized by a positive zeta potential of about 20 at a pH of about 7, while unmodified MaSp fibers have a zeta potential of about -20 at a pH of about 7.

[0115] In some embodiments, derivatized (e.g., amino-derivatized) porous MaSp fibers have a zeta potential that is at least 50%, at least 70%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 700%, at least 800%, and at least 1000% greater than the zeta potential of unmodified porous MaSp fibers.

[0116] In some embodiments, derivatized (e.g., carboxy-derivatized) porous MaSp fibers are characterized by a zeta potential value of -20 to -100 at a pH of about 7. As illustrated herein, carboxylated MaSp fibers are characterized by a zeta potential value of about -40 at a pH of about 7, and unmodified MaSp fibers have a zeta potential value of about -20 at a pH of about 7.

[0117] In some embodiments, derivatized (e.g., carboxy-derivatized) porous MaSp fibers have a zeta potential that is at least 50%, at least 70%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 700%, at least 800%, and at least 1000% lower than that of unmodified porous MaSp fibers. Those skilled in the art will understand that the exact zeta potential is determined by the pH and the loading of functional groups into the porous MaSp fibers (i.e., the w / w ratio).

[0118] In some embodiments, the derivatized MaSp fibers retain the porosity of the original (e.g., underivativeized) MaSp fibers. In some embodiments, the derivatized MaSp fibers are at least 10 m 2 It is characterized by a BET surface area of ​​ / g.

[0119] SEM images of derivatized MaSp fibers are shown in Figures 3A and 3B. As shown in Figures 3A and 3B, both the MaSp protein and the mutated MaSp protein exhibit a highly porous structure, thus demonstrating the excellent porosity of the derivatized MaSp fibers (e.g., at least 10 m). 2 The porosity (determined by the BET surface area per gram) is sequence-independent. Furthermore, the porosity of derivatized MaSp fibers is substantially retained compared to that of unmodified MaSp fibers.

[0120] In some embodiments, the derivatized porous MaSp fibers of the present invention comprise a functional moiety covalently bonded to a polymer, a linker, and a chelate moiety or any combination thereof. In some embodiments, the functional moiety is covalently bonded to a linker, the linker being as described herein. In some embodiments, the functional moiety is covalently bonded to a chelate moiety, the chelate moiety being one of a metal chelate group, a metal oxide chelate group or a combination thereof, the metal chelate group and metal oxide chelate group being as described herein.

[0121] In some embodiments, the derivatized porous MaSp fibers of the present invention include dyes or pigments bonded to a functional moiety, which is as described herein. In some embodiments, the dyes or pigments are bonded to the functional moiety via covalent or non-covalent bonds. In some embodiments, the dyes or pigments are bonded to the functional moiety via hydrogen bonds, van der Waals interactions, electrostatic interactions, pp stacking, or any combination thereof.

[0122] Examples of dyes, though not limited to them, include anionic dyes (e.g., Congo Red, Alizarin Pure Blue B, Acid Red 88, Trypan Blue), cationic dyes (e.g., methine dyes, anthraquinone dyes, azo dyes, Coomassi, Methylene Blue), and neutral dyes (e.g., Neutral Orange RL, Neutral Red GRL, Neutral Gray 2BL), Brilliant Carmine 6B, Lake Red C, Watching Red, Diazo Yellow, Hansa Yellow, Phthalocyanine Blue, Phthalocyanine Green, Alkali Blue, and Aniline Black, or any combination thereof. Other neutral or charged organic dyes are well known in the art.

[0123] The inventors have successfully implemented the PEI and / or amino-modified derivatized fibers of the present invention in the hair coloring compositions described in the Examples section, along with various dyes.

[0124] In some embodiments, the composition of the present invention comprises the derivatized porous MaSp fiber of the present invention and further components optionally selected from addition polymers, dyes and / or pigments.

[0125] In some embodiments, the composition comprises MaSp fibers in amounts of 0.01% to 50%, 0.01% to 1%, 1% to 5%, 5% to 10%, 10% to 15%, 15% to 20%, and 20% to 50% (w / w) (including any range between these) and further components.

[0126] In some embodiments, this simple is suitable for concentrations of 0.001%~95% (w / w), 0.005%~95% (w / w), 0.009%~95% (w / w), 0.01%~95% (w / w), 0.05%~95% (w / w), 0.09%~95% (w / w), 0.1%~95% (w / w), 0.5%~95% (w / w), 0.9%~95% (w / w), 1%~95% (w / w), and 5%~95%. %(w / w), 10%~95%(w / w), 15%~95%(w / w), 20%~95%(w / w), 30%~95%(w / w), 50%~95%(w / w), 0.01%~80%(w / w) ), 0.05%~80%(w / w), 0.09%~80%(w / w), 0.1%~80%(w / w), 0.5%~80%(w / w), 0.9%~80%(w / w), 1%~80%(w / w) , 5%~80%(w / w), 10%~80%(w / w), 15%~80%(w / w), 20%~80%(w / w), 30%~80%(w / w), 50%~80%(w / w), 0.001%~ 50%(w / w), 0.005%~50%(w / w), 0.009%~50%(w / w), 0.01%~95%(w / w), 0.01%~50%(w / w), 0.05%~50%(w / w) This includes derivatized porous MaSp fibers in the following concentrations: 0.09%~50%(w / w), 0.1%~50%(w / w), 0.5%~50%(w / w), 0.9%~50%(w / w), 1%~50%(w / w), 5%~50%(w / w), 10%~50%(w / w), 15%~50%(w / w), 20%~50%(w / w), or 30%~50%(w / w) (including any range in between).

[0127] In some embodiments, the composition of the present invention essentially consists of the derivatized porous MaSp fibers of the present invention and optionally further components described herein. In some embodiments, at least 80%, at least 85%, at least 90%, at least 92%, at least 95%, at least 97%, at least 99%, and at least 99.9% by weight of the composition (including any range in between) consists of the derivatized porous MaSp fibers of the present invention and optionally further components described herein.

[0128] In some embodiments, the derivatized porous MaSp fiber essentially consists of any one of the derivatized porous MaSp fibers described herein. In some embodiments, the derivatized porous MaSp fiber of the present invention substantially lacks further fibers (e.g., derivatized fibers) and / or further polymers and / or further organic or inorganic materials or particles.

[0129] In some embodiments, at least 80%, at least 85%, at least 90%, at least 92%, at least 95%, at least 97%, at least 99%, and at least 99.9% (including any range in between) by weight of the derivatized porous MaSp fibers of the present invention consist of any one of the derivatized porous MaSp fibers described herein.

[0130] In some embodiments, the derivatized porous MaSp fibers and / or compositions of the present invention are stable. In some embodiments, the derivatized porous MaSp fibers of the present invention are said to be stable if the derivatized fibers retain their physical and / or chemical properties, and / or are chemically and / or physically stable when dispersed in solution and / or during long-term storage under ambient storage conditions and / or when exposed to heat at temperatures up to 300°C, up to 200°C, up to 100°C, up to 80°C, and up to 60°C (including any range in between).

[0131] In some embodiments, the compositions of the present invention are stable when the derivatized porous MaSp fibers of the present invention are stably bound to further components (for example, the compositions are chemically stable when dispersed in solution and / or during long-term storage under ambient storage conditions and / or when exposed to heat at temperatures up to 300°C, up to 200°C, up to 100°C, up to 80°C, and up to 60°C (including any range in between)).

[0132] In some embodiments, ambient conditions include exposure to any one of inert chemicals such as a solvent (organic solvent and / or aqueous solvent, where the solvent is inert, i.e., does not chemically react with any of the components of the composition), thermal exposure to temperatures up to 300°C, 200°C, 100°C, 80°C, and 60°C (including any range in between), exposure to UV / vis radiation (and / or electromagnetic radiation, IR radiation, microwave radiation, etc.), and exposure to moisture and / or atmospheric gases. In some embodiments, ambient conditions include repeated exposure to inert chemicals. In some embodiments, ambient conditions include exposure to temperatures below the melting point and / or decomposition point of any of the components of the composite (e.g., MaSp fibers or derivatized MaSp fibers). Those skilled in the art will understand that the precise definition of ambient storage conditions may include further parameters or conditions well known in the art.

[0133] In some embodiments, the compositions and / or derivatized porous MaSp fibers of the present invention are said to be stable if they substantially maintain their structure and physical properties (e.g., mechanical stability, porosity, tensile strength, etc.) and chemical properties (wettability, zeta potential, hydrophobic / hydrophilic, reactivity) and / or if further components remain in contact with or bound to the derivatized MaSp fibers of the present invention (e.g., substantially do not disintegrate).

[0134] In some embodiments, the compositions and / or derivatized MaSp fibers of the present invention are said to be stable if they substantially maintain their chemical composition.

[0135] In some embodiments, the compositions and / or derivatized MaSp fibers of the present invention are substantially chemical and / or physically stable for at least 1 month (m), at least 2 m, at least 6 m, at least 12 m, at least 2 years (y), at least 3 y, at least 10 y (including any range in between), substantially as described below herein. In some embodiments, the compositions and / or derivatized MaSp fibers of the present invention are substantially stable for the periods described herein under ambient storage conditions.

[0136] composite material In another embodiment, a composite material is provided comprising the derivatized porous MaSp fibers of the present invention bonded to a metal, a metal salt, or metal oxide particles, or any combination thereof. In some embodiments, the metal oxide particles are bonded to the derivatized porous MaSp fibers via coordination bonds, electrostatic interactions, or both.

[0137] In some embodiments, the composite material of the present invention comprises a derivatized porous MaSp fiber of the present invention bonded to a metallic component selected from a metal, a metal salt, or a metal oxide, or any combination thereof, wherein the metallic component is in the form of particulate matter or distinct atoms, and the metallic component is in an elemental state or an oxidized state. In some embodiments, the metallic component comprises a metal and / or a metal salt, wherein the metallic component comprises a first metal and / or a second metal of the present invention.

[0138] In some embodiments, the composite material of the present invention comprises derivatized porous MaSp fibers of the present invention bonded to a metal component described herein via one or more chelating agents of the present invention.

[0139] In some embodiments, a metal component is coordinately bonded to a derivatized MaSp fiber via one or more chelating agents of the present invention, where one or more chelating agents of the present invention are covalently bonded to a functional portion of the present invention (e.g., a polymer described herein). In some embodiments, the composite material of the present invention comprises a derivatized porous MaSp fiber of the present invention bonded to a metal component, where the derivatized porous MaSp fiber comprises a plurality of chelating agents covalently bonded to a polymer described herein. In some embodiments, the chelating agents are as described herein (e.g., (i) a metal chelating group that can bond to a metal or a salt thereof, (ii) a metal oxide chelating group, or both).

[0140] In some embodiments, the composite material of the present invention comprises metal oxide particles bonded to porous MaSp fibers derivatized via a chelating agent. In some embodiments, the chelating agent is a metal oxide chelating group described herein. In some embodiments, the metal oxide particles are complexed with the chelating agent. In some embodiments, the metal oxide particles are stably bonded to the derivatized porous MaSp fibers (for example, the composite material is chemically stable when dispersed in solution and / or during long-term storage under ambient storage conditions and / or when exposed to heat at temperatures up to 300°C, up to 200°C, up to 100°C, up to 80°C, and up to 60°C (including any range in between)).

[0141] In some embodiments, ambient conditions include exposure to any one of inert chemicals such as a solvent (organic solvents and / or aqueous solvents, where the solvent is inert, i.e., does not chemically react with any of the components of the composite), thermal exposure to temperatures up to 300°C, 200°C, 100°C, 80°C, and 60°C (including any range in between), exposure to UV / vis radiation (and / or electromagnetic radiation, IR radiation, microwave radiation, etc.), and exposure to moisture and / or atmospheric gases. In some embodiments, ambient conditions include repeated exposure to inert chemicals. In some embodiments, ambient conditions include exposure to temperatures below the melting point and / or decomposition point of any of the components of the composite (e.g., MaSp fibers or derivatized MaSp fibers). Those skilled in the art will understand that the precise definition of ambient storage conditions may include further parameters or conditions well known in the art.

[0142] In some embodiments, the composites of the present invention are said to be stable if they substantially maintain their structure and physical properties (e.g., mechanical stability, porosity, tensile strength, conductivity, etc.) and chemical properties (wettability, zeta potential, hydrophobic / hydrophilicity, reactivity) and / or if the metallic components remain in contact with or bonded to the derivatized MaSp fibers of the present invention (e.g., substantially do not disintegrate), where substantially means as described herein.

[0143] In some embodiments, the composite material of the present invention is said to be stable if the composite material substantially maintains its chemical composition.

[0144] In some embodiments, the composites of the present invention are substantially chemical and / or physically stable for at least 1 month (m), at least 2 m, at least 6 m, at least 12 m, at least 2 years (y), at least 3 y, at least 10 y (including any range in between), substantially as described below herein. In some embodiments, the composites of the present invention are substantially stable under ambient storage conditions for the periods described herein.

[0145] In some embodiments, the composite material of the present invention comprises derivatized MaSp fibers of the present invention doped with a metal component. In some embodiments, the metal component is homogeneously dispersed in the composite material (e.g., on or within porous nanofibrils).

[0146] In some embodiments, the metal oxide is selected from the group consisting of titanium oxide, aluminum oxide, iron(II / III) oxide, zirconium oxide, zinc oxide, silicon oxide, or any mixture thereof. In some embodiments, the metal oxide chelating group has affinity and / or selectivity for titanium oxide. In some embodiments, the chelating agent has affinity and / or selectivity for metal oxide particles. In some embodiments, the chelating agent has affinity and / or selectivity for titanium oxide particles. In some embodiments, the metal oxide particles are as described above herein.

[0147] In some embodiments, the derivatized porous MaSp fibers include functional moieties covalently bonded to metal oxide chelate groups. In some embodiments, the metal oxide chelate groups are covalently bonded to the functional moieties via linkers, where the linkers are as described herein. In some embodiments, the metal oxide chelate groups are covalently bonded to the functional moieties via PGA linkers. In some embodiments, each PGA chain is covalently bonded to multiple metal oxide chelate groups.

[0148] In some embodiments, the functionalized portion of a derivatized porous MaSp fiber covalently bonded to a metal oxide chelate group is given by formula 4: [ka] This is expressed as follows, where the dashed line represents any combination, and each n is an independent integer between 1 and 10000.

[0149] In some embodiments, the metal oxide chelate group comprises carboxy and / or hydroxyl groups. In some embodiments, the metal oxide chelate group comprises small molecules and / or polymers. In some embodiments, the metal oxide chelate group is a monodentate ligand, a bidentate ligand, a tridentate ligand, and a tetradentate ligand. In some embodiments, the metal oxide chelate group comprises a ligand having affinity and / or selectivity for titania. In some embodiments, the metal oxide chelate group comprises a ligand containing one or more carboxyl groups and optionally one or more hydroxyl groups. In some embodiments, the metal oxide chelate group comprises a ligand, i.e., a cyclic polydentate ligand or a linear ligand.

[0150] In some embodiments, the metal oxide chelate group is a polymer containing carboxyl side chain groups and / or hydroxyl side chain groups (such as PVA, polyacrylates, polyglycolates (including any mixture or copolymer thereof)).

[0151] Non-exclusive examples of metal oxide chelating groups include, but are not limited to, salicylic acid, phosphonic acid, hydroxamic acid, malonic acid, pyrogallol, 5-hydroxy-1,4-naphthoquinone, quinone, or any combination thereof. Other metal oxide chelating groups with affinity for titania are well known in the art.

[0152] In some embodiments, the metal oxide chelate group includes an oxidized tyrosine side chain (e.g., dihydroxyphenyl or quinone). In some embodiments, the derivatized MaSp fiber includes at least one oxidized tyrosine residue (e.g., in the form of dihydroxyphenyl or quinone). Those skilled in the art will understand that oxidized tyrosine can be obtained, for example, by reacting a MaSp fiber with tyrosinase to obtain at least a portion of the oxidized tyrosine residue (e.g., in the form of dihydroxyphenyl or quinone).

[0153] In some embodiments, the metal oxide chelate group is bonded to a polymer, which is described herein. In some embodiments, the metal oxide chelate group is covalently bonded to a polymer, which is described herein. In some embodiments, the chelate portion is covalently bonded to a functional group of a derivatized MaSp fiber.

[0154] In some embodiments, the composite material of the present invention comprises a derivatized porous MaSp fiber containing a plurality of metal oxide chelate groups bonded thereto, where at least a portion (e.g., 20-99%, 20-30%, 30-40%, 40-60%, 60-80%, 80-90%, 90-99% (including any range in between)) of the metal oxide chelate groups are bonded to a metal oxide (e.g., the metal oxide particles of the present invention).

[0155] Those skilled in the art will understand that there are many options for covalent bonding of the chelate portion (e.g., a metal oxide chelate group) to the polymer. For example, an aminated chelate portion can be bonded to a carboxylated MaSp fiber. Alternatively, a carboxylated chelate portion can be bonded to an aminated MaSp fiber. Furthermore, an aminated chelate portion can be bonded to a polymer containing a carboxyl group or a carbonyl group (e.g., PGA). Alternatively, a carboxylated chelate portion can be bonded to a polymer containing a hydroxyl group or an amino group (e.g., polylysine or PEI). The inventors have succeeded in synthesizing an aminated MaSp fiber bonded to PGA, in which PGA is further bonded to a metal oxide chelate group (salicylic acid) as shown herein.

[0156] In some embodiments, the composite material of the present invention comprises a derivatized porous MaSp fiber containing a functional moiety covalently bonded to a metal chelating group. In some embodiments, the metal chelating group is covalently bonded to the functional moiety via a linker, where the linker is as described herein. In some embodiments, the metal chelating group is covalently bonded to the functional moiety via a PGA linker. In some embodiments, each PGA chain is covalently bonded to one of the metal chelating groups described above.

[0157] In some embodiments, the composite material of the present invention comprises a derivatized porous MaSp fiber containing a plurality of metal chelating groups bonded thereto, where at least a portion of the metal chelating groups (e.g., 20-99%, 20-30%, 30-40%, 40-60%, 60-80%, 80-90%, 90-99% (including any range in between)) is bonded to a metal, where the metal is as described herein. In some embodiments, at least a portion of the metal chelating groups is bonded to Pd or a salt thereof (e.g., Pd(acetate)2, PdCl2). In some embodiments, at least a portion of the metal chelating groups is bonded to the metal in an elemental state (e.g., ground state, also known as the 0 oxidation state). In some embodiments, at least a portion of the metal chelating groups is bonded to the metal in an oxidation state (e.g., +2). In some embodiments, the metal chelating groups are bonded to a metal that is at least partially reduced.

[0158] In some embodiments, the composition of the present invention comprises a derivatized fiber of the present invention doped with a metal (the first and / or second metal described herein), where the metal is coordinately bonded to a polymer containing multiple metal chelating groups. In some embodiments, the composition of the present invention comprises Pd complexed with a polyglutaraldehyde containing multiple metal chelating groups (such as IDA), where the polyglutaraldehyde is covalently bonded to the derivatized MaSp fiber of the present invention.

[0159] In some embodiments, polymers containing multiple metal chelate groups that complex with a metal or metal cation include: [ka] This is expressed as follows, where k is between 10 and 10000, M represents a transition metal (e.g., Pd or a salt thereof), and m and n each independently represent an integer between 0 and 10.

[0160] In another aspect of the present invention, the metal atoms (or first metal) complexed by the polymer of the present invention are further bonded to further metal atoms, where the further metal atoms are the same or different. In some embodiments, the metal atoms complexed by the polymer of the present invention are further bonded to multiple metal atoms, thereby forming aggregates. In some embodiments, the first metal (e.g., Pd) complexed by the polymer of the present invention is uniformly dispersed on the outer surface of the derivatized fibers of the present invention. In some embodiments, the first metal (e.g., Pd) forms a metal layer (or first metal layer) on the derivatized fibers of the present invention. In some embodiments, the metal layer (e.g., first metal layer and / or second metal layer) is 1 to 10 atoms thick (including any range between them). In some embodiments, the first metal (e.g., Pd) is in colloidal form. In some embodiments, the first metal (e.g., Pd) is in particle form ranging from 1 to 500 nm.

[0161] In some embodiments, the first metal, which is either in the form of particles or distinct atoms, forms aggregation centers suitable for bonding the second metal thereon. In some embodiments, Pd (in its elemental state) complexed with the polymer of the present invention as described herein forms aggregation centers to enable the deposition of the second metal (e.g., Cu). In some embodiments, the second metal is deposited by electroless plating. In some embodiments, the first metal can facilitate the electroless plating of the second metal.

[0162] In some embodiments, the second metal (e.g., Cu) is in colloidal form. In some embodiments, the second metal (e.g., Cu) is in the form of particles in the ranges of 1–500 nm, 1–10 nm, 10–50 nm, 50–100 nm, 100–200 nm, and 200–500 nm (including any range in between).

[0163] In some embodiments, the second metal is either in the form of particulate matter or as distinct atoms, where the distinct atoms are in an elemental state or an oxidized state. In some embodiments, the second metal, which is either in the form of particulate matter or as distinct atoms, is homogeneously dispersed on the derivatized fibers of the present invention. In some embodiments, the first metal and / or the second metal is in the form of a homogeneous layer on the derivatized fibers of the present invention. In some embodiments, the first metal and / or the second metal is in an amorphous state or a crystalline state (e.g., forming substantially crystalline particles). In some embodiments, at least a portion of the first metal and / or the second metal is in a crystalline state.

[0164] "Uniform" or "homogeneous" is intended to mean a distribution of different sizes (or thicknesses) within a range of, for example, ±60%, ±50%, ±40%, ±30%, ±20%, or less than ±10% (including any value in between).

[0165] In some embodiments, the term “layer” refers to a substantially uniform thickness of a substantially homogeneous material. In some embodiments, the shell comprises a single layer or multiple layers.

[0166] In some embodiments, the derivatized MaSp fiber is deposited on or bonded to a second metal (e.g., Cu) on a first metal (e.g., Pd). 0The invention includes a layer of ) in which the first metal is complexed with polyglutaraldehyde covalently bonded to the MaSp fiber. In some embodiments, the first metal (e.g., Pd) is chelated or complexed with a metal chelate moiety (e.g., IDA). In some embodiments, the metal chelate moiety is covalently bonded to the polyglutaraldehyde as illustrated herein. The inventors have derived the MaSp fiber into Cu 0 We succeeded in doping with it, and here the derivatized MaSp-based fibers contain PGA bonded to an IDA metal chelate group.

[0167] In some embodiments, the first metal and / or the second metal are in the form of particulate matter or separate atoms as described herein, where the first metal and the second metal are independently in an elemental state or an oxidized state.

[0168] In some embodiments, a metal layer (e.g., a first metal layer) in contact with or bonded to a polymer layer forms multiple aggregation sites for a second metal. In some embodiments, the first metal has a high affinity for the coating polymer of the polymer layer. In some embodiments, the first metal is bonded to the coating polymer.

[0169] In some embodiments, the second metal is bonded or aggregated on top of the first metal. In some embodiments, the second metal is in an elemental state. In some embodiments, the second metal is in an oxidized state (e.g., +1 or +2). In some embodiments, the second metal forms a layer on top of the first metal layer. In some embodiments, the second metal has affinity for the first metal. In some embodiments, the second and first metals are in the form of a layered structure, where each metal layer is separated. In some embodiments, the second and first metals are mixed together in a metal layer. In some embodiments, the second and first metals are mixed together to form a single metal layer on top of a polymer layer. In some embodiments, the fiber is at least partially coated with either the first and second metals or a combination of the first and second metals.

[0170] In some embodiments, the first metal and optionally the second metal are transition metals as described herein. Transition metals are well known in the art and are referred to as d-electron-containing metals.

[0171] In some embodiments, the first metal has a reduction potential suitable for chemical reduction. In some embodiments, the first and second metals are compatible with electroless plating methods. In some embodiments, the first metal can directly reduce the second metal.

[0172] In some embodiments, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, and at least 99% (including any range in between) of the chelating agent is bonded to metal oxide particles.

[0173] In some embodiments, the w / w ratio of derivatized porous MaSp fibers to metal oxide particles is 0.01:1~100:1, 0.01:1~10:1, 0.01:1~0.05:1, 0.05:1~0.1:1, 0.1:1~0.2:1, 0.2:1~0.3:1, 0.3:1~0.4:1, 0.4:1~0.5:1, The ranges are 0.5:1 to 0.7:1, 0.7:1 to 0.9:1, 0.5:1 to 1:1, 0.9:1 to 1:1, 1:1 to 1.5:1, 1.5:1 to 2:1, 2:1 to 3:1, 3:1 to 5:1, 5:1 to 7:1, 7:1 to 10:1, 10:1 to 30:1, 30:1 to 50:1, and 50:1 to 100:1 (including any range in between).

[0174] In some embodiments, the metal oxide particles are characterized by particle sizes of 25-5000 nm, 25-50 nm, 50-100 nm, 100-150 nm, 150-200 nm, 200-300 nm, 300-500 nm, 500-1000 nm, 1000-2000 nm, 2000-3000 nm, 3000-4000 nm, and 4000-5000 nm (including any range in between).

[0175] In some embodiments, the composite material is stable for at least 1 month (m), at least 2m, at least 3m, at least 4m, at least 5m, at least 6m, at least 7m, at least 8m, at least 9m, at least 10m, and at least 12m (including any range in between).

[0176] In some embodiments, the metal oxide within the metal oxide particles is in an amorphous state. In some embodiments, the metal oxide within the metal oxide particles is in a crystalline state. In some embodiments, at least a portion of the metal oxide within the metal oxide particles is in an amorphous state.

[0177] In some embodiments, the composite material is characterized by enhanced dispersibility in aqueous and / or organic solutions compared to the original metal oxide particles (e.g., those without MaSp fibers). The composite material of the present invention, containing titanium oxide particles having a particle size greater than 300 nm, showed significantly improved dispersibility in aqueous and / or organic solutions compared to the original titania particles. Aqueous dispersions containing titanium oxide particles bonded to salicylic acid or PVA-derivativeized MaSp fibers (e.g., exemplary composite materials of the present invention) showed superior stability compared to control dispersions containing underivativeized MaSp fibers. Furthermore, the composite material containing titanium oxide particles bonded to salicylic acid-derivativeized MaSp fibers exhibited excellent dispersibility (e.g., the formation of stable dispersions), where the w / w ratio of titanium oxide particles to derivatized MaSp fibers was approximately 1:1.

[0178] In some embodiments, the aqueous dispersion formed from the composite material of the present invention further contains a surfactant. Exemplary surfactants used for forming a stable dispersion include, but are not limited to, TRITON and sodium dodecyl sulfate (SDS).

[0179] In some embodiments, the composite material is characterized by enhanced absorption of UV radiation compared to a control. In some embodiments, the control is a derivatized or underivatized MaSp fiber that does not contain metal oxide particles.

[0180] In some embodiments, the composites of the present invention are characterized by electrical conductivity (also referred to herein as "conductive"), where the composites are or include a derivatized MaSp system bonded to or doped with a metal (e.g., a metal in its elemental state as described herein). In some embodiments, the metal induces or enhances the conductivity of the composites of the present invention. In some embodiments, a derivatized MaSp system bonded to or doped with a metal (e.g., a metal in its elemental state as described herein) is characterized by improved conductivity compared to a derivatized MaSp fiber that does not contain a metal. In some embodiments, a polymer reinforced with derivatized MaSp fibers (e.g., reinforced by 1-30%, 1-10%, 10-20%, 20-30% (including any value in between) relative to the total weight of the composites) is characterized by improved conductivity compared to the polymer in its original state (e.g., without derivatized MaSp fibers).

[0181] In some embodiments, the original MaSp fibers (e.g., underivativeized fibers) and / or derivatized MaSp fibers that do not contain metals are substantially nonconductive. In some embodiments, the original MaSp fibers (e.g., underivativeized fibers) and / or derivatized MaSp fibers that do not contain metals are at least 10 13 ohm*cm, at least 10 14 ohm*cm, at least 10 15 It is characterized by a volume resistivity of ohm*cm (including any range between them).

[0182] In some embodiments, the conductivity of the composite is at least 50%, at least 100%, at least 200%, at least 500%, at least 1000%, at least 10000%, and at least 10000000000% (including any range in between) greater than the conductivity of the original derivatized MaSp fibers (e.g., lacking the bonded metal).

[0183] In some embodiments, the conductivity of the composite material is at least 10, at least 100, at least 1,000, at least 10,000, at least 100,000, at least 1,000,000, and at least 10,000,000 times greater (including any range in between) than the conductivity of the original derivatized MaSp fibers.

[0184] In some embodiments, the composite material of the present invention is 10 12 ~10 -5 ohm / m, 10 12 ~10 10 ohm / m, 10 10 ~10 8 ohm / m, 10 8 ~10 6 ohm / m, 10 6 ~10 4 ohm / m, 10 4 ~10 2 ohm / m, 10 2 ~1 ohm / m, 1~10 -5 ohm / m, 1-10 -2 ohm / m, 10 -2 and 10 -3 ohm / m, 10 -3 ~10 -5 It is characterized by a resistivity of ohm / m (including any range in between). In some embodiments, this resistivity is called the electrical resistivity normalized with respect to the length of the sample.

[0185] In some embodiments, derivatized MaSp fibers bonded to or doped with metal are characterized by antimicrobial activity that substantially reduces or prevents the amount of microorganisms on the surface of the derivatized MaSp fibers or on the surface of further polymers reinforced with the derivatized MaSp fibers.

[0186] method In another embodiment, a method for synthesizing a derivatized porous MaSp fiber of the present invention, wherein the porous MaSp fiber is given formula I: [ka] Or formula II: [ka] A method is provided for obtaining a derivatized porous MaSp fiber, comprising the step of reacting with a reagent represented by the formula, where R comprises the functional part of the present invention, A is selected from substituted or unsubstituted aryl, heteroaryl, and alkyl (including any combination thereof), each R1 independently comprises one of hydrogen, alkyl, hydroxy, or alkoxy (including any combination thereof), the wavy bond represents a linker or bond, and the reacting step comprises conditions sufficient to covalently bond the functional part to tyrosine of a porous MaSp fiber. In some embodiments, the linker is as described herein.

[0187] In some embodiments, at least one R1 contains a leaving group. In some embodiments, each R1 is independently hydrogen or -OC 1~10 It contains any one of the alkyl groups. In some embodiments, A contains a substituted or unsubstituted aromatic ring. In some embodiments, A contains a substituted or unsubstituted phenyl group.

[0188] In some embodiments, the linker is substituted or unsubstituted C 1~10 Contains alkyl.

[0189] In some embodiments, the method for synthesizing the derivatized porous MaSp fibers of the present invention includes the steps of preparing underivativeized porous MaSp fibers and reacting them with a diazonium salt represented by formula I, thereby covalently bonding the reagent to the tyrosine of the MaSp fibers via diazo bonds. In some embodiments, the diazonium salt is a nitrite salt represented by formula 5: [ka] It is synthesized by reaction with an amine, where R and A are as described herein. In some embodiments, the diazonium salt is synthesized by reaction in an aqueous solution (e.g., aqueous buffer). In some embodiments, the pH of the aqueous solution is 1 to 7, 1 to 3, 5 to 7, 3 to 5 (including any range in between).

[0190] In some embodiments, underivativeized porous MaSp fibers are reacted with a reagent of formula I in an aqueous solution (or aqueous suspension) or a dispersion containing a polar organic solvent. In some embodiments, sufficient conditions for the covalent bonding of the functional moiety of the porous MaSp fibers to tyrosine include a pH of the aqueous solution between 3 and 10, 5 and 7, 3 and 5, 7 and 10 (including any range in between). In some embodiments, sufficient conditions for the covalent bonding of the functional moiety of the porous MaSp fibers to tyrosine include a temperature below 15°C, below 10°C, below 5°C, and below 3°C (including any range in between).

[0191] In some embodiments, the method for synthesizing the derivatized porous MaSp fibers of the present invention includes the steps of preparing underivativeized porous MaSp fibers and reacting them with a reagent represented by formula II, thereby covalently bonding the reagent to the tyrosine of the MaSp fibers via silyl bonds (C-Si bonds).

[0192] In some embodiments, underivativeized porous MaSp fibers are reacted with the reagent of formula II in an aqueous solution (or aqueous suspension) or dispersion containing a polar organic solvent. In some embodiments, conditions sufficient for the covalent bonding of the functional moiety of the porous MaSp fibers to tyrosine include a pH of the aqueous solution between 6 and 12, 5 and 7, 6 and 10, 7 and 10, 7 and 9, and 9 and 12 (including any range in between). In some embodiments, conditions sufficient for the covalent bonding of the functional moiety of the porous MaSp fibers to tyrosine include a temperature below 15°C, below 10°C, below 5°C, and below 3°C (including any range in between).

[0193] In some embodiments, the method further comprises reacting a functional moiety (e.g., an amine or carboxyl group) with a polymer having reactivity to the functional moiety, thereby covalently bonding the polymer to a porous MaSp fiber, wherein the functional moiety comprises an amine or carboxyl group. In some embodiments, the functional moiety (e.g., an amine) is reacted with the carboxyl group of the polymer by adding a coupling agent (such as HATU, HOBt, CDI, EDC, NHS, or a mixture thereof). In some embodiments, the functional moiety (e.g., carboxyl group) is reacted with the amine group of the polymer by adding a coupling agent (such as HATU, HOBt, CDI, EDC, NHS, or a mixture thereof). Various coupling agents are well known in the art, along with reaction conditions that are sufficiently precise for the coupling of amines and carboxyls.

[0194] In some embodiments, a functional moiety (e.g., an amine) is reacted with a carbonyl group (ketone or aldehyde) of the polymer to form an imine. In some embodiments, the method further includes a step of reducing the imine to an amine, for example, with sodium borohydride or by hydrogen reduction with a suitable catalyst. Sufficient reaction conditions for imine bond formation and further imine-amine reduction are well known in the art.

[0195] In some embodiments, the method further includes the step of reacting the polymer (e.g., PGA) with a chelating agent that is reactive with the polymer to obtain a chelating agent covalently bonded to the polymer. In some embodiments, the method further includes the step of reacting the polymer (e.g., PGA) with a chelating agent to obtain a chelating agent covalently bonded to the polymer via imine or amide bonds, wherein the polymer contains a carbonyl or carboxyl group, and the chelating agent contains an amine group.

[0196] Porous MaSp fibers According to some embodiments, the present invention provides a composition comprising derivatized porous MaSp-based fibers. In some embodiments, the derivatized MaSp-based fibers are present at a concentration of 0.1% to 90% by total weight.

[0197] In some embodiments, the porous MaSp-based fibers comprise at least one MaSp-based fiber. In some embodiments, the at least one MaSp-based fiber is present at a concentration of 0.1% to 25%, 0.1% to 20%, 0.1% to 15%, 0.5% to 30%, 1% to 30%, 5% to 30%, or 10% to 30% (including any range therebetween) by total weight.

[0198] In some embodiments, the porous MaSp-based fibers are MaSp-based polymers in the form of particles having a size in the range of 0.5 μm to 1.5 μm. In some embodiments, the MaSp-based fibers are insoluble polymers. In some embodiments, the porous MaSp-based fibers have a DSC pattern showing an endothermic peak in the range of at least 200°C to 280°C. In some embodiments, the porous MaSp-based fibers have an amide peak in the range of 1615 cm -1 ~1635 cm -1 as characterized by FTIR analysis.

[0199] According to some embodiments, a composition comprising a MaSp-based polymer (e.g., a synthetic MaSp-based polymer) is provided, wherein the MaSp-based polymer has at least one property evaluation selected from: a) being an insoluble polymer; b) being in the form of particles having a size in the range of 0.5 μm to 1.5 μm; c) having a DSC pattern showing an endothermic peak in the range of at least 200°C to 280°C; and d) having an amide peak in the range of 1615 cm -1 ~1638 cm -1 as measured by FTIR analysis.

[0200] In some embodiments, when determined by differential scanning calorimetry (DSC), the MaSp-based fiber has a decomposition temperature (T d ) of 280°C to 350°C, 290°C to 350°C, 300°C to 350°C, 310°C to 350°C, 320°C to 350°C, 280°C to 330°C, 290°C to 330°C, 300°C to 330°C, 310°C to 330°C or 320°C to 330°C (including any range therebetween). Each possibility represents a separate embodiment of the invention.

[0201] In some embodiments, when determined by DSC, the MaSp-based fiber has a glass transition temperature (T g ) of 200°C to 250°C, 210°C to 250°C, 220°C to 250°C, 230°C to 250°C, 200°C to 240°C, 210°C to 240°C, 220°C to 240°C, 230°C to 240°C, 200°C to 230°C or 210°C to 230°C (including any range therebetween). Each possibility represents a separate embodiment of the invention.

[0202] In some embodiments, when determined by DSC, the MaSp-based fiber has a T g of 260°C to 320°C, 270°C to 320°C, 280°C to 320°C, 290°C to 320°C, 260°C to 310°C, 270°C to 310°C, 280°C to 310°C or 290°C to 310°C (including any range therebetween). Each possibility represents a separate embodiment of the invention.

[0203] In some embodiments, the MaSp-based fiber is characterized by a DSC pattern showing an endothermic peak of at least 280°C to 350°C, 290°C to 350°C, 300°C to 350°C, 310°C to 350°C, 280°C to 330°C, 290°C to 330°C, 300°C to 330°C, 310°C to 330°C or 320°C to 330°C (including any range therebetween). Each possibility represents a separate embodiment of the invention.

[0204] As used herein, the term "decomposition temperature (T d )" refers to the temperature at which decomposition occurs. Thermal decomposition is a process of extensive chemical species change caused by heat.

[0205] The term "glass transition temperature (T)" as used herein refers to the glass transition temperature (T) used in this specification. g The term ) refers to a material that is an elastic and viscous amorphous liquid (T>T g ) from brittle, glassy amorphous solid (T <T g This refers to the temperature at which the transition occurs to liquid-glass. This liquid-glass transition (or simply glass transition) is a reversible transition. g ) is generally the melting temperature (T) of the crystalline state of the material if it exists. m It is lower than ).

[0206] According to some embodiments, porous MaSp-based fibers are provided, comprising a synthetic MaSp-based polymer in the form of particles. In some embodiments, the particles have sizes in the range of 0.5 μm to 1.5 μm, 0.7 μm to 1.5 μm, 0.8 μm to 1.5 μm, 0.9 μm to 1.5 μm, 0.5 μm to 1 μm, 0.7 μm to 1 μm, 0.8 μm to 1 μm, 0.9 μm to 1 μm, 0.5 μm to 1.3 μm, 0.5 μm to 1.2 μm, 0.7 μm to 1.3 μm, 0.7 μm to 1.2 μm, or 0.9 μm to 1.2 μm (including any range in between).

[0207] In some embodiments, the MaSp fiber comprises or consists of an insoluble MaSp polymer. In some embodiments, the insoluble MaSp polymer is in the form of particles. In some embodiments, the insoluble MaSp polymer is insoluble in organic solvents. In some embodiments, the insoluble MaSp polymer is insoluble in aqueous solutions. The terms “MaSp polymer” and “MaSp fiber” as used herein are interchangeable herein.

[0208] As used herein, the term “insoluble” refers to a material that does not dissolve when exposed to an excess of solvent but can be dispersed to varying degrees. In some embodiments, “insoluble” refers to a material that is soluble in the solvent at concentrations of less than 10%, less than 5%, less than 2%, or less than 1%. In some embodiments, “insoluble” refers to a material that can be partially dissolved in the solvent at concentrations of less than 0.01% by weight. Solvents according to the present invention include organic solvents and aqueous solutions. In some embodiments, the solvent includes an aqueous surfactant solution. In some embodiments, the solvent includes an aqueous urea solution.

[0209] In some embodiments, the MaSp fiber is characterized by a specified differential scanning calorimetry (DSC) pattern. In some embodiments, “DSC pattern” is intended to refer to the location of the peak. In some embodiments, “peak” is intended to refer to the exothermic peak. Throughout this specification, “location of the peak” or “peak position” refers to the peak along the temperature axis in the thermogram pattern, and in some embodiments, it may refer to the location of the peak at any peak intensity. Those skilled in the art will understand that the data obtained in a DSC measurement is determined in part by the equipment used and the environmental conditions (e.g., humidity) at the time the measurement is performed.

[0210] In some embodiments, the MaSp-based polymer is characterized by a DSC pattern showing an endothermic peak in the range of at least 200°C to 280°C. In some embodiments, the disclosed composition is characterized by a DSC pattern showing an endothermic peak in the range of at least 200°C to 270°C, 200°C to 260°C, 200°C to 250°C, 210°C to 280°C, 212°C to 280°C, 215°C to 280°C, 216°C to 280°C, 220°C to 280°C, 210°C to 250°C, 212°C to 250°C, 215°C to 250°C, 216°C to 250°C, 220°C to 250°C, 210°C to 245°C, 210°C to 242°C, or 215°C to 245°C (including any range in between).

[0211] In some embodiments, the MaSp polymer is characterized by a DSC pattern exhibiting endothermic peaks that are at least 5°C to 100°C, at least 10°C to 100°C, at least 15°C to 100°C, at least 12°C to 100°C, at least 25°C to 100°C, at least 5°C to 80°C, at least 10°C to 80°C, at least 15°C to 80°C, at least 12°C to 80°C, at least 25°C to 80°C, at least 5°C to 50°C, at least 10°C to 50°C, at least 15°C to 50°C, at least 12°C to 50°C, or at least 25°C to 50°C lower than the DSC pattern of the corresponding composition containing (MaSp) fibers.

[0212] In some embodiments, MaSp polymers lack DSC peaks in the range of approximately -100°C to approximately 190°C. In some embodiments, the disclosed compounds lack DSC peaks in the range of approximately -100°C to approximately 25°C. In some embodiments, the disclosed compositions are characterized by a DSC pattern that shows the absence of exothermic peaks in the range of at least 40°C to 70°C.

[0213] In some embodiments, the MaSp polymer lacks a DSC peak in the range of approximately -100°C to approximately -50°C. In some embodiments, the disclosed compound lacks a DSC peak in the range of approximately -50°C to approximately 0°C. In some embodiments, the disclosed compound lacks a DSC peak in the range of approximately -0°C to approximately -25°C.

[0214] In some embodiments, the MaSp polymer yielded 1615 cm³ when measured by FTIR analysis. -1 ~1635cm -1 It is characterized by having an amide peak in the range of . In some embodiments, the disclosed composition has a peak of 1620 cm when measured by FTIR analysis. -1 ~1635cm -1 , 1620cm -1 ~1630cm -1 , 1621cm -1 ~1630cm -1 or 1620cm -1~1625cm -1 It is characterized by having an amide peak within the range (including any range within that range).

[0215] In some embodiments, the MaSp polymer yielded 1700 cm³ when measured by FTIR analysis. -1 ~1800cm -1 The peak is missing within this range.

[0216] In one embodiment, the MaSp polymer of the present invention assembles by self-assembly. "Self-assembly" means that the monomers of the present invention, i.e., synthetic spider silk proteins, spontaneously combine with each other in an energetically favorable manner under normal physiological conditions or at room temperature to form a macromolecular structure having the properties described herein. Furthermore, the MaSp polymer of the present invention is extremely resilient and, once assembled, can withstand extreme chemical attacks such as solubilization in a 10% surfactant solution and boiling for at least one hour.

[0217] "Tenacity" or "tensile strength" refers to the weight of a filament that can withstand before fracture. The maximum specific stress produced by a tensile test to fracture a material is typically the one in a filament, yarn, or fabric. According to specific embodiments, the MaSp polymer of the present invention has a tensile strength of approximately 100-3000 MPa (MPa = N / mm²). 2 It has a tensile strength of approximately 300-3000 MPa, approximately 500-2700 MPa, approximately 700-2500 MPa, approximately 900-2300 MPa, approximately 1100-2000 MPa, approximately 1200-1800 MPa, approximately 1300-1700 MPa, or approximately 1400-1600 MPa. More specifically, it is approximately 1500 MPa.

[0218] "Toughness" refers to the energy required to break a MaSp-based polymer. This is the area under the stress-strain curve, and is sometimes called "breaking energy" or "work of fracture." According to certain embodiments, the MaSp-based polymer of the present invention has a toughness of approximately 20-1000 MJ / m². 3 , about 50~950MJ / m 3, approximately 100 - 900 MJ / m 3 , approximately 120 - 850 MJ / m 3 , approximately 150 - 800 MJ / m 3 , approximately 180 - 700 MJ / m 3 , approximately 180 - 750 MJ / m 3 , approximately 250 - 700 MJ / m 3 , approximately 280 - 600 MJ / m 3 , approximately 300 - 580 MJ / m 3 , approximately 310 - 560 MJ / m 3 , approximately 320 - 540 MJ / m 3 or approximately 350 - 520 MJ / m 3 , most particularly approximately 350 - 520 MJ / m 3 has toughness.

[0219] "Elasticity" refers to the property of a body that tends to return to its original size and shape after deformation. Plasticity, i.e., deformation without recovery, is the opposite of elasticity. The recoverable, i.e., elastic deformation in the molecular arrangement of MaSp-based polymers is possible by the elongation (reorientation) of interatomic and intermolecular structural bonds. Conversely, the breaking of intermolecular bonds and reformation into new stabilized positions causes non-recoverable, i.e., plastic deformation.

[0220] "Elongation" refers to an increase in length expressed as a ratio or fraction of the initial length.

[0221] "Fineness" means the average diameter of MaSp-based polymers or filaments (e.g., biofilaments), which is usually expressed in microns (micrometers).

[0222] MaSp-based fibers The terms “large bottle gland spidoin protein” and “spirulina protein” are used synonymously throughout this specification and encompass all known large bottle gland spidoin proteins, which are typically abbreviated as “MaSp” or, in the case of the garden spider, “ADF.” These large bottle gland spidoin proteins are generally of two types, 1 and 2. These terms further include non-natural proteins disclosed herein that have a high degree of identity and / or similarity to at least the repeating regions of known large bottle gland spidoin proteins. Further preferred spider silk proteins include MaSp2, MiSp, MiSp2, AcSp, FLYS, FLAS, and flagellar gland-derived proteins.

[0223] As used herein, the terms “repeating region,” “repeating sequence,” or “repeat” refer to recombinant protein sequences derived from repeat units that occur multiple times naturally in the spider silk amino acid sequence (e.g., in the MaSp-1 protein). Those skilled in the art will understand that the primary structure of a spider silk protein is considered to consist primarily of a series of small variations of a unit repeat. These unit repeats in naturally occurring proteins are often different from one another; that is, there is little or no exact overlap of unit repeats along the length of the protein. In some embodiments, the synthetic spider silk of the present invention is prepared such that the primary structure of the protein contains many exact repeats of a single unit repeat. In further embodiments, the synthetic spider silk of the present invention contains many repeats of one unit repeat along with many repeats of a second unit repeat. Such structures are similar to typical block copolymers. Alternatively, unit repeats of several different sequences may be combined to obtain a synthetic spider silk protein with properties suitable for a particular application. As used herein, the term “direct repeat” refers to a tandem repeat (head-tail arrangement) having similar repeats. In another embodiment, the repeats used to form the synthetic spider silk of the present invention are direct repeats. In some embodiments, such repeats do not exist in nature (i.e., they are not naturally occurring amino acid sequences).

[0224] An exemplary sequence containing a repeating sequence is ADF-4:(SEQ ID NO: 1). In some embodiments, the synthetic repeating sequence of the present invention is based on one or more repeating sequences derived from ADF-4(SEQ ID NO: 1) (e.g., having a high degree of identity as defined herein). As used herein, the term “based on” refers to a sequence having a high degree of homology to the repeating sequence.

[0225] In some embodiments, each repeat sequence includes up to 60 amino acids, up to 55 amino acids, up to 50 amino acids, up to 49 amino acids, up to 48 amino acids, up to 47 amino acids, up to 46 amino acids, up to 45 amino acids, up to 44 amino acids, up to 43 amino acids, up to 42 amino acids, up to 41 amino acids, up to 40 amino acids, up to 39 amino acids, up to 38 amino acids, up to 37 amino acids, up to 36 amino acids, or up to 35 amino acids, where each possibility represents a separate embodiment of the present invention. In some embodiments, each repeat sequence includes 5 to 60 amino acids, 10 to 55 amino acids, 15 to 50 amino acids, 20 to 45 amino acids, 25 to 40 amino acids, an acid, 25 to 39 amino acids, or 28 to 36 amino acids, where each possibility represents a separate embodiment of the present invention. In some embodiments, each repeat sequence comprises 30–40 amino acids, 31–39 amino acids, 32–38 amino acids, 33–37 amino acids, and 34–36 amino acids, where each possibility represents a distinct embodiment of the present invention. In further embodiments, each repeat sequence comprises 35 amino acids.

[0226] In some embodiments, the repeating region is independently the amino acid sequence (X1) shown in formula 10. Z X2GPGGYGPX3X4X5GPX6GX7GGX8GPGGPGX9X 10 The formula includes such that X1 is independently A or G in each case.

[0227] In some embodiments, (X1) Z At least 50% of X is A, Z is an integer between 5 and 30, X2 is S or G, X3 is G or E, X4 is G, S or N, X5 is Q or Y, X6 is G or S, X7 is P or R, X8 is Y or Q, X9 is G or S, X 10 It is either S or G.

[0228] In another embodiment, the repeat region of the MaSP1 protein includes the amino acid sequence shown in SEQ ID NO: 2 (SGPGGYGPGSQGPSGPGGYGPGGPGSS). In yet another embodiment, the repeat region of the MaSP1 protein includes the amino acid sequence shown in SEQ ID NO: 3 (AAAAAAAASGPGGYGPGSQGPSGPGGYGPGGPGSS).

[0229] In another embodiment, homologs of repeat regions of the MaSP1 protein are provided that share at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology with SEQ ID NO: 1.

[0230] In another embodiment, the homologue shares at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology with sequence number 2.

[0231] In another embodiment, the repeat region of the MaSP1 protein has the amino acid sequence shown in SEQ ID NO: 1.

[0232] In another embodiment, the MaSP1 protein includes a single N-terminal region selected from the group consisting of SEQ ID NO: 4 (MSYYHHHHHHDYDIPTTENLYFQGAMDPEFKGLRRRAQLV), SEQ ID NO: 5 (MSYYHHHHHHDYDIPTTENLYFQGAMDPEFKGLRRRAQLVRPLSNLDNAP), SEQ ID NO: 6 (MSYYHHHHHHDYDIPTTENLYFQGAMDPEFKGLRRRAQLVDPPGCRNSARAGSS), or any functional homologue, variant, derivative, or fragment thereof. In another embodiment, the C-terminal region homologue shares at least 70% homology with any one of SEQ ID NOs: 4-6.

[0233] In another embodiment, the MaSP1 protein further comprises a single C-terminal region selected from the group consisting of SEQ ID NO: 7 (VAASRLSSPAASSRVSSAVSSLVSSGPTNGAAVSGALNSLVSQISASNPGLSGCDALVQALLELVSALVAILSSASIGQVNVSSVSQSTQMISQALS), SEQ ID NO: 8 (GPSGPGAYGPSPSASASVAASRLSSPAASSRVSSAVSSLVSSGPTNGAAVSGALNSLVSQISASNPGLSGCDALVQALLELVSALVAILSSASIGQVNVSSVSQSTQMISQALS), or any functional homologue, variant, derivative, fragment, or mutant thereof. In another embodiment, the N-terminal region homologue shares at least 70% homology with SEQ ID NOs: 7-8.

[0234] In some embodiments, the MaSp fiber comprises a mixture of proteins disclosed in International Publication No. 2017025964.

[0235] In some embodiments, the MaSp fiber contains a mutated protein obtained by expressing a mutated nucleic acid sequence.

[0236] In some embodiments, the MaSP1 protein further comprises at least one tag sequence. Non-limiting examples of tags that can be used in the present invention include His tags, HA tags, and T7 tags. Those skilled in the art are well aware of other suitable tags or other fusion partners.

[0237] MaSp-based fibers containing microbial interaction peptides According to some embodiments, the MaSp system fiber contains a microbial interaction protein. In some embodiments, the microbial interaction protein is a virus-binding receptor.

[0238] In some embodiments, the MaSp fiber contains a peptide that can interact with microorganisms (e.g., viruses). In some embodiments, the MaSp fiber contains a peptide that can bind to microorganisms (e.g., viruses).

[0239] In some embodiments, the microbial interaction protein is a virus-binding receptor. In some embodiments, the virus-binding receptor is a receptor used for viral entry. In some embodiments, the virus interaction protein is a host cell surface component recognized by the virus. In some embodiments, viral recognition is recognized as an entry and exit point for cell entry.

[0240] In some embodiments, the virus is a coronavirus. In some embodiments, the coronavirus is an alpha-coronavirus. In some embodiments, the coronavirus is 229E or NL63. In some embodiments, the coronavirus is a beta-coronavirus. In some embodiments, the coronavirus is OC43, HKU1, or MERS-CoV. In some embodiments, the coronavirus is SARS-CoV-1. In some embodiments, the coronavirus is SARS-CoV-2.

[0241] In some embodiments, the virus-binding receptor is angiotensin-converting enzyme 2 (ACE2). In some embodiments, the virus is a coronavirus and the virus-binding receptor is angiotensin-converting enzyme 2 (ACE2).

[0242] In some embodiments, ACE2 is a mutated ACE2. In some embodiments, the mutated ACE2 is a cleavage-resistant ACE2 mutant.

[0243] Table 1 shows two non-restrictive sequences that can be used as virus-binding receptors. [Table 1]

[0244] In some embodiments, the MaSp fiber of the present invention comprises an N-terminus, a repeating region (e.g., 24 repeats), and a C-terminus. The repeats define a specific structure, and the C-terminus is important for self-assembly. In some embodiments, a microbial interaction protein (e.g., a virus-binding receptor) can be inserted into the N-terminal region, which has little to no effect on the structure (e.g., porosity) of the resulting protein.

[0245] A non-restrictive example of a polynucleotide sequence encoding a virus-binding receptor is GCCAAATGTATCCACTACAAGAAATTCAGAATGGTGAGGAACAGGCCAAGACATTTTTGGACAAGTTTAACCACGAAGCCGAAGACCTGTTCTATCAAAGTTCAGGACTGGGGAAGGGCGACTTCAGG (Sequence ID 11).

[0246] Since the N-terminus of MaSp fibers is a polylinker with multiple restriction sites, any restriction site that appears in the polylinker but not in the insert may be used for introducing a polynucleotide sequence.

[0247] Table 2 provides a list of possible restriction enzymes for introducing polynucleotide sequences encoding virus-binding receptors into the N-terminal region of MaSp filaments. [Table 2]

[0248] As used herein, “amino acids” refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimes that function similarly to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code and those that are later modified, such as hydroxyproline, γ-carboxyglutamic acid, and O-phosphoserine. “Amino acid analogs” refer to compounds that have the same basic chemical structure as naturally occurring amino acids, namely hydrogen, a carboxyl group, an amino group, and an alpha carbon bonded to an R group, such as homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. Such analogs have a modified R group or a modified peptide skeleton but retain the same basic chemical structure as naturally occurring amino acids. “Amino acid mimes” refer to compounds that have a different structure from the general chemical structure of amino acids but function similarly to naturally occurring amino acids. In this specification, amino acids may be referred to by either their commonly known three-letter or one-letter symbols as recommended by the IUPAC-IUB Biochemical Nomenclature Commission.

[0249] An "amino acid sequence" or "peptide sequence" is the order in which amino acid residues linked by peptide bonds are located within a chain of peptides and proteins. This sequence is generally reported from the N-terminus, which contains a free amino group, to the C-terminus, which contains a free carboxyl group. While amino acid sequences are often called peptides, or protein sequences, when they represent the primary structure of a protein, proteins are defined as amino acid sequences folded into a specific three-dimensional structure and typically undergo post-translational modifications such as phosphorylation, acetylation, glycosylation, sulfhydryl bond formation, and cleavage. Therefore, the terms "amino acid sequence" or "peptide sequence" must be distinguished from the term "protein."

[0250] As used herein, “isolated” or “substantially purified” means, in the context of the synthetic spider silk amino acid sequences or nucleic acid molecules encoding them as illustrated by the present invention, that the amino acid sequences or polynucleotides have been removed from their natural environment or altered from their natural state. Thus, “isolated” does not necessarily reflect the degree to which the amino acid sequences or nucleic acid molecules are purified. However, it will be understood that such molecules that are purified to some extent are “isolated.” If a molecule does not exist in its natural environment, i.e., does not exist naturally, then the molecule is “isolated,” regardless of where it exists. For example, an amino acid sequence or polynucleotide that does not exist naturally in humans is “isolated,” even if it is present in humans.

[0251] The terms “isolated” or “substantially purified,” when applied to amino acid sequences or nucleic acids, mean that the amino acid sequence or nucleic acid essentially does not contain other cellular components to which it is naturally associated. It may exist in a homogeneous state, or in either dry or aqueous solution. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high-performance liquid chromatography. The amino acid sequence or nucleic acid, which is the main species present in the formulation, is substantially purified.

[0252] In some embodiments, the repeat is a homolog, variant, or derivative of the repeat region of the MaSp1 protein or a fragment thereof. In some embodiments, the repeat is a homolog, variant, or derivative of the repeat region of the ADF-4 protein or a fragment thereof.

[0253] As used herein in relation to "functional homologues, variants, derivatives, or fragments," the term "functionality" refers to an amino acid sequence having a biological function or activity identified by a specified functional assay. More specifically, the specified functional assay is the formation of self-assembling fibers in cells expressing the functional homologue, variant, derivative, or fragment.

[0254] An amino acid sequence or nucleic acid sequence is a homolog of the corresponding amino acid sequence or nucleic acid if it is determined that the homology is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, or at least 99%.

[0255] The homology used herein may be determined based on the percentage of identity between two amino acids (peptides) or DNA sequences. Generally, the two sequences to be compared are aligned to obtain the greatest correlation between them. The alignment of the two sequences is examined, the number of positions that give an exact amino acid (or nucleotide) correspondence between the two sequences is determined, and the percentage of identity is obtained by dividing this number by the total length of the alignment and multiplying by 100. This percentage of identity may be determined over the entire length of the sequences being compared, which is particularly suitable for sequences of the same or very similar length that are highly homologous, or it may be determined over a shorter specified length, which is more suitable for sequences of unequal length, i.e., sequences with a lower level of homology. Methods for comparing the identity of two or more sequences are well known in the art. Therefore, for example, the percentage of identity between two amino acid sequences and the percentage of identity between two polynucleotide sequences may be determined using programs available in version 9.1 of the Wisconsin Sequence Analysis Package, such as the GAP and BESTFIT programs. BESTFIT uses Smith and Waterman's "local homology" algorithm to find the best single region of similarity between two sequences. BESTFIT is better suited for comparing two polypeptides or polynucleotide sequences of different lengths, and the program assumes that the shorter sequence represents a portion of the longer sequence. In comparison, GAP aligns the two sequences and finds the "maximum similarity" according to Needleman and Wunsch's algorithm. GAP is better suited for comparing sequences of approximately the same length, and alignment is expected to be across the entire length. Preferably, the "gap weight" and "length weight" parameters used in each program are 50-3 for polynucleotide sequences and 12-4 for polypeptide sequences, respectively. Preferably, the percentages of identity and similarity are determined when the two sequences being compared are optimally aligned.

[0256] The terms “identical,” “substantial identity,” “substantial homology,” or “identical percentage” refer to two or more amino acid or nucleic acid sequences that, when measured using the BLAST or BLAST2.0 sequence comparison algorithm with the default parameters described below, or by manual alignment and visual inspection, are either the same amino acid residue or nucleotide, or have a specified percentage of the same amino acid residue or nucleotide (i.e., approximately 60% identity, or at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or 99% identity, when compared and aligned for maximum match across a comparison window or specified region). Such sequences are then said to be “substantial identity.” This definition may also refer to, or apply to, complementary sequences of test sequences. This definition includes sequences with deletions and / or additions, as well as sequences with substitutions. Preferred algorithms can account for gaps, etc.

[0257] Typically, one sequence acts as a reference sequence for sequence comparison, and the test sequence is compared to it. When using a sequence comparison algorithm, the test and reference sequences are input into a computer, subsequence coordinates are specified if necessary, and sequence algorithm program parameters are specified. Preferably, default program parameters can be used, or other parameters can be specified. The sequence comparison algorithm then calculates the degree of sequence identity of the test sequence to the reference sequence based on the program parameters.

[0258] Naturally, the present invention further encompasses amino acid sequences comprising 2 to 70 repeats of any one variant of SEQ ID NOs: 1, 2, or 3. As used herein, the terms “variant” or “substantially similar” include amino acid or nucleotide sequences that differ from a specifically identified sequence in which one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, or 25) amino acid residues or nucleotides are deleted, substituted, or added. Variants may be naturally occurring allelic variants or non-natural variants. Variants or substantially similar sequences refer to amino acid sequences or nucleic acid fragments that may be characterized by a degree of identity between those amino acid or nucleotide sequences and the amino acid or nucleotide sequences described herein, as determined by common algorithms used in state-of-the-art art. Preferred fragments of amino acids or nucleic acids have an amino acid or nucleotide sequence having at least about 40 or 45% sequence identity, preferred about 50 or 55% sequence identity, more preferably about 60 or 65% sequence identity, more preferably about 70 or 75% sequence identity, more preferably about 80 or 85% sequence identity, and even more preferably about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity when compared to a reference sequence.

[0259] In one embodiment, the MaSp polymer is a fiber.

[0260] In one embodiment, the MaSp polymer consists of monomers. In one embodiment, multiple monomers are arranged within nanofibrils. In one embodiment, multiple nanofibrils are arranged within or constitute a fiber. In one embodiment, the monomers or nanofibrils within the MaSp polymer or fiber have a diameter of 4 to 16 nm. In one embodiment, the monomers or nanofibrils within the MaSp polymer or fiber have a diameter of 6 to 14 nm. In one embodiment, the monomers or nanofibrils within the MaSp polymer or fiber have a diameter of 8 to 12 nm. In one embodiment, the fiber or MaSp polymer has a diameter of 70 to 450 nm. In one embodiment, the protein fiber or MaSp polymer has a diameter of 80 to 350 nm. In one embodiment, the fiber or MaSp polymer has a diameter of 80 to 300 nm. In one embodiment, the fiber or MaSp polymer has a diameter of 150 to 250 nm. In one embodiment, the fiber or MaSp polymer is arranged as a coil. In one embodiment, a single fiber or a single MaSp-based polymer is arranged as a coil. In one embodiment, the coil has a diameter of 5 to 800 micrometers. In one embodiment, the coil has a diameter of 5 to 500 micrometers. In one embodiment, the coil has a diameter of 5 to 30 micrometers. In one embodiment, the coil has a diameter of 5 to 20 micrometers. In one embodiment, the fiber or MaSp-based polymer has a length of 5 to 800 micrometers. In one embodiment, the fiber or MaSp-based polymer has a length of 30 to 300 micrometers.

[0261] In one embodiment, the fiber or MaSp polymer is branched. In one embodiment, the fiber or MaSp polymer contains 1 to 10 branches. In one embodiment, the fiber or MaSp polymer does not contain carbohydrates. In one embodiment, the fiber or MaSp polymer is not glycosylated. In one embodiment, the fiber or MaSp polymer does not contain fats or fatty acids. In one embodiment, the fiber or MaSp polymer does not contain phosphorus. In one embodiment, the fiber or MaSp polymer does not contain further non-MaSp proteins. In one embodiment, the fiber or MaSp polymer does not contain further polymers (e.g., synthetic polymers, non-MaSp peptides, non-MaSp proteins). In one embodiment, the fiber or MaSp polymer substantially does not contain further polymers. In one embodiment, "does not contain" means "lacks" or essentially "lacks."

[0262] In one embodiment, the length-to-diameter aspect ratio of the fiber, i.e., the MaSp-based polymer, is at least 1:10. In one embodiment, the length-to-diameter aspect ratio of the fiber or MaSp-based polymer is at least 1:10 to 1:1500. In one embodiment, the length-to-diameter aspect ratio of the fiber or MaSp-based polymer is at least 1:50 to 1:1000. In one embodiment, the length-to-diameter aspect ratio of the fiber or MaSp-based polymer is at least 1:100 to 1:1200. In one embodiment, the length-to-diameter aspect ratio of the fiber or MaSp-based polymer is at least 1:100 to 1:1000. In one embodiment, the length-to-diameter aspect ratio of the fiber or MaSp-based polymer is at least 1:500 to 1:1000.

[0263] As used herein, the terms derivative and functional derivative mean the amino acid sequences of the present invention having any insertions, deletions, substitutions, and modifications.

[0264] Naturally, as used herein, the term “insertion” means the addition of any number of amino acid residues to the sequence of the present invention, specifically 1 to 50 amino acid residues, more specifically 20 to 1 amino acid residues, and more specifically 1 to 10 amino acid residues. Most specifically, these are 1, 2, 3, 4, 5, 6, 7, 8, 9 to 10 amino acid residues. Furthermore, the amino acid sequence of the present invention may be extended at its N-terminus and / or C-terminus by a variety of identical or different amino acid residues.

[0265] An amino acid "substitution" is the result of replacing one amino acid with another amino acid having similar structural and / or chemical properties, i.e., a conservative amino acid substitution. Amino acid substitutions may be made based on the similarity of the residues involved in terms of polarity, charge, solubility, hydrophobicity, hydrophilicity, and / or amphiphilicity. For example, nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine; polar, neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine; positively charged (basic) amino acids include arginine, lysine, and histidine; and negatively charged (acidic) amino acids include aspartic acid and glutamic acid.

[0266] In another embodiment, the repeat sequence of the present invention has 17 or fewer, 16 or fewer, 15 or fewer, 14 or fewer, 13 or fewer, 12 or fewer, 11 or fewer, 10 or fewer, 9 or fewer, 8 or fewer, or 7 or fewer amino acid substitutions in any one of sequence numbers 2 or 3. In one embodiment, the repeat sequence of the present invention has at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 amino acid substitutions in any one of sequence numbers 2 or 3.

[0267] Those skilled in the art with respect to amino acid sequences will recognize that individual substitutions, deletions, or additions to an amino acid, nucleic acid, peptide, polypeptide, or protein sequence that alter, add, or delete a single amino acid or a small proportion of amino acids in the encoded sequence are “conservatively modified variants” in which the alteration results in the substitution of an amino acid with a chemically similar amino acid. Tables of conservative substitutions that provide functionally similar amino acids are well known in the art. Such conservatively modified variants also include, but are not excluded from, the polymorphic variants, interspecific homologs, and alleles of the present invention.

[0268] For example, substitutions may be made in which an aliphatic amino acid (G, A, I, L, or V) is replaced with another member of the following groups, i.e., substitutions in which one polar residue is replaced with another, such as replacing arginine with lysine, glutamic acid with aspartic acid, or glutamine with asparagine. Each of the following eight groups includes other exemplary amino acids in which substitutions are conserved: 1) alanine (A), glycine (G); 2) aspartic acid (D), glutamic acid (E); 3) asparagine (N), glutamine (Q); 4) arginine (R), lysine (K); 5) isoleucine (I), leucine (L), methionine (M), valine (V); 6) phenylalanine (F), tyrosine (Y), tryptophan (W); 7) serine (S), threonine (T); and 8) cysteine ​​(C), methionine (M).

[0269] A conservative nucleic acid substitution is a nucleic acid substitution that yields the conservative amino acid substitution defined above.

[0270] The amino acid sequence variant of the present invention may have at least 80% sequence similarity, at least 85% sequence similarity, 90% sequence similarity, or at least 95%, 96%, 97%, 98%, or 99% sequence similarity at the amino acid level with the repeat unit represented by either one of SEQ ID NOs: 2 or 3.

[0271] The amino acid sequence of the present invention may contain 2 to 70 repeats of SEQ ID NO: 1 or 3 or any fragment thereof. A “fragment” constitutes an amino acid or DNA sequence fraction of a specific region. A peptide sequence fragment is at least one amino acid shorter than the specific region, and a DNA sequence fragment is at least one base pair shorter than the specific region. The fragment may be cleaved at the C-terminus, N-terminus, or both. The amino acid fragment may contain at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 24, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, or at least 34 amino acids from SEQ ID NO: 1 or 3.

[0272] The amino acid sequence mutants of the present invention are characterized by the exchange of one or more amino acids (point mutants) for one or more other amino acids, with a maximum of about 10 amino acids. These are the result of corresponding mutations at the DNA level that lead to different codons.

[0273] Furthermore, the present invention relates to derivatives of the amino acid sequence of the present invention. Derivatives of the amino acid sequence of the present invention are derivatized by functional groups such as amino, hydroxyl, mercapto, or carboxyl groups, for example, by glycosylation, acylation, amidation, or esterification. In glycosylated derivatives, oligosaccharides are usually bonded to asparagine, serine, threonine, and / or lysine. Acylated derivatives are acylated particularly with naturally occurring organic or inorganic acids, such as acetic acid, phosphoric acid, or sulfuric acid, and it is usually particularly at the N-terminal amino or hydroxyl group of tyrosine or serine, respectively. Esters are esters of naturally occurring alcohols, such as methanol or ethanol. Further derivatives are salts, particularly pharmaceutically acceptable salts, such as metal salts, such as alkali metal and alkaline earth metal salts, such as sodium, potassium, magnesium, calcium, or zinc salts, or ammonium salts formed with ammonia or suitable organic amines, such as triethylamine, hydroxy-lower alkylamines, such as 2-hydroxyethylamine.

[0274] In some embodiments, the silk protein of the present invention lacks post-translational modifications.

[0275] In some embodiments, the silk proteins of the present invention are biodegradable. This property can always be important, for example, in the medical field, when the silk proteins are intended for in vivo use where biological degradation is desired. This property may be applied to suture materials and wound closure and covering systems.

[0276] According to several embodiments, the MaSp fibers of the present invention are produced using an expression vector containing a suitable nucleic acid sequence, where the nucleic acid sequence is under the expression control of an operablely linked promoter and optionally a regulatory sequence. Exemplary expression systems, such as the expression system disclosed in PCT / Israeli Patent No. 2020 / 050752, are known in the art.

[0277] In some embodiments, MaSp proteins self-assemble to form a predetermined structure. In some embodiments, MaSp proteins exist as a network. In some embodiments, MaSp proteins exist as a complex. In some embodiments, MaSp proteins induce predetermined secondary structures, such as β-turns, γ-turns, β-sheets, and α-helix structures.

[0278] In some embodiments, the MaSp proteins or MaSp polymers used interchangeably herein are in the form of fibers. As used herein, “fiber” means a thin string of fibrous material consisting of two or more filaments twisted together. “Filament” means an elongated, thread-like object or structure of indeterminate length ranging from microscopic to more than one mile. Specifically, synthetic spider silk filaments are microscopic and proteinaceous. “Biofilament” means a filament made from a protein containing recombinant spider silk protein. In some embodiments, the term “fiber” does not include unstructured aggregates or precipitates.

[0279] In some embodiments, the protein fiber is characterized by a size of at least one dimension (e.g., diameter, length). For example, but not limited to, the diameter of the fiber may be 10 nm to 1 μm, 20 to 100 nm, or 10 to 50 nm.

[0280] In some embodiments, the fiber consists of nanofibrils. In some embodiments, the nanofibrils have a diameter of, for example, 1 nm, about 2 nm, about 3 nm, about 4 nm, about 5 nm, about 6 nm, about 7 nm, about 8 nm, about 9 nm, about 10 nm, about 11 nm, about 12 nm, about 13 nm, about 14 nm, about 15 nm, about 16 nm, about 17 nm, about 18 nm, about 19 nm, about 20 nm, about 21 nm, about 22 nm, about 23 nm, about 24 nm, about 25 nm, about 26 nm, about 27 nm, about 28 nm, about 29 nm, about 30 nm, about 31 nm, about 32 nm, about 33 nm, about 34 nm, about 35 nm, about 36 nm, about 37 nm, about 38 nm, about 40 nm, about 42 nm, about 44 nm, about 46 nm, about 48 nm or about 50 nm (including any value or range in between). In one embodiment, the nanofibril has a diameter of 3 to 7 nm. In another embodiment, the nanofibril has a diameter of 4 to 6 nm.

[0281] In some embodiments, the disclosed fiber lengths are 1–200 μm, 10–100 μm, 100–500 μm, or 200–500 μm.

[0282] In some embodiments of any one of the embodiments described herein, the disclosed fibers (e.g., particles) are characterized by a porous structure. In some embodiments, the porous structure is characterized by at least 30% (e.g., 30-99%) porosity. In some embodiments, the porous structure is characterized by at least 50% (e.g., 50-99%) porosity. In some embodiments, the porous structure is characterized by at least 60% (e.g., 60-99%) porosity. In some embodiments, the porous structure is characterized by at least 70% (e.g., 70-99%) porosity. In some embodiments, the porous structure is characterized by at least 80% (e.g., 80-99%) porosity. In some embodiments, the porous structure is characterized by at least 90% (e.g., 90-99%) porosity. In some embodiments, the porous structure is characterized by about 90% porosity.

[0283] In this specification, the term "porosity" refers to the percentage of volume of voids in a material (e.g., a "sponge-like" material). In another embodiment, porosity is measured according to the voids or lumens within a divided surface area relative to the total surface area (porous and non-porous).

[0284] In some embodiments, the porous structure of the disclosed fibers allows for efficient absorption of water onto the fiber surface. That is, although not bound by any particular theory, this remarkable discovery can be explained in terms of the structure of the disclosed fibers and their porosity, which are strictly distinct from natural spider silk found in nature.

[0285] In some of the embodiments described herein, the disclosed fibers are characterized by having an average diameter of nanoscale.

[0286] In some embodiments, the disclosed fibers are characterized by having an average diameter in the range of 1 to 50 nm. In some such embodiments, the average diameter is in the range of 3 to 50 nm. In some such embodiments, the average diameter is in the range of 5 to 50 nm. In some such embodiments, the average diameter is in the range of 1 to 40 nm. In some such embodiments, the average diameter is in the range of 1 to 30 nm. In some such embodiments, the average diameter is in the range of 5 to 40 nm.

[0287] In some embodiments, the MaSp fibers contain multiple pores. In some embodiments, the porous MaSp fibers contain multiple fibrils (e.g., nanofibrils) as illustrated below this specification (Figures 4A and 4B). In some embodiments, the MaSp fibers are in the form of particles as described below this specification. In some embodiments, the MaSp fibers are as described below this specification. In some embodiments, the composition contains multiple MaSp fibers. In some embodiments, the multiple MaSp fibers contain fibers having different chemical compositions and / or different molecular weights (MW).

[0288] As further illustrated in the examples below, in some embodiments, the plurality of disclosed fibers may be in the form of a self-assembled structure or matrix. In some embodiments, this matrix can be made suitable for biomaterial applications.

[0289] In some embodiments, this matrix is suitable for cell growth and for maintaining or promoting cell viability, as further demonstrated below in this specification.

[0290] In some embodiments, the term "self-assembled" refers to a structure that results from a self-assembly process (e.g., a spontaneous self-assembly process) based on a series of associative chemical reactions between at least two domains of the fiber, where the associative groups on one domain are in a sufficiently proximate state and are oriented to enable a configurational association with another domain. In other words, associative interactions imply proximity that results in the adhesion of domains of one or more fibers to each other. In some embodiments, the adhered domains are not parallel to each other. Configurations are also contemplated where there are three or more domains of a self-assembled structure that engage in different planes.

[0291] In some embodiments, the density of the self-assembled fibers (e.g., about 80% void space) is notable in that it ranges from 0.1 g / cm 3 to 0.4 g / cm 3 or from 0.2 g / cm 3 to 0.3 g / cm 3 In an exemplary embodiment, the density of the self-assembled fibers is about 0.26 g / cm 3 .

[0292] The study of surface wettability at nanoscale spatial resolution and high temporal resolution is an emerging field from both theoretical and practical aspects. The disclosed fibers exhibited a high degree of surface wettability with an excellent fluid absorption capacity compared to their volume and weight.

[0293] In some embodiments, the polymer is hydrophobic. In some embodiments, the polymer is UV curable.

[0294] In some embodiments, the disclosed composite material is biologically stable. In some embodiments, the disclosed composite material is biocleavable. In some embodiments, the disclosed composite material is biodegradable.

[0295] In some embodiments, the term “biological stability” refers to a compound or polymer that remains unchanged under physiological conditions (e.g., not degraded in vivo and therefore non-biodegradable or non-biologically cleavable).

[0296] In some embodiments, the term “biodegradable” refers to a substance that can be broken down into degradation products under physiological and / or environmental conditions. Such physiological and / or environmental conditions include, for example, hydrolysis (degradation by hydrolytic cleavage), enzymatic catalysis (enzymatic degradation), and mechanical interactions. The term typically refers to a substance that degrades under these conditions such that 50% by weight of the substance degrades within a period of less than one year.

[0297] In some embodiments, the term “biodegradable” as used in the context of embodiments of the present invention also encompasses the term “bioresorbable,” which refers to a substance that is bioabsorbed by a host organism under physiological conditions, i.e., a substance that breaks down into degradation products that become metabolites of the host organism’s biochemical system.

[0298] antibacterial composition The present invention is based in part on the discovery that MaSp fibers and / or derivatized MaSp fibers of the present invention can be used to provide an enhanced antimicrobial effect by various means, including but not limited to the incorporation of microbial interaction proteins and / or at least one antimicrobial agent onto MaSp fibers having a nanoporous structure characterized by a large surface area.

[0299] The present invention is further partly based on providing MaSp-based fibers having enhanced wettability, thereby enabling increased incorporation of droplets containing microorganisms onto the fibers. Furthermore, the present invention is further partly based on providing MaSp-based fibers that thereby incorporate antimicrobial agents or metals (e.g., by adsorption or covalent bonding).

[0300] According to several embodiments, the present invention provides a composition comprising a MaSp fiber containing at least one antimicrobial agent and / or a derivatized MaSp fiber of the present invention. According to several embodiments, the antimicrobial agent and the MaSp fiber are bonded via hydrogen bonds, van der Waals bonds, or both. According to several embodiments, the antimicrobial agent and the MaSp fiber are bonded via hydrogen bonds, π-π electrostatic bonds, or both. According to several embodiments, the antimicrobial agent and the MaSp fiber and / or a derivatized MaSp fiber of the present invention are bonded via covalent bonds.

[0301] In some embodiments, the antimicrobial agent is selected from the group consisting of reactive oxygen species (ROS) sources, carboxylic acids, quaternary amines, bactericides, transition metals, electrophilic reactive groups, oxidizing agents, antimicrobial polymers, or any combination thereof.

[0302] In some embodiments, ROS includes oxygen-based reactants. In some embodiments, ROS includes peroxides, singlet oxygen, peroxides, superoxides, hydroxyl radicals, and alpha oxygen or combinations thereof. ROS contributes to the oxidation of the extracellular envelope of microorganisms (e.g., bacteria and / or viruses).

[0303] In some embodiments, the ROS source includes a photosensitizer. In some embodiments, the photosensitizer can generate ROS in situ by activation with a light source. In some embodiments, the photosensitizer includes rose bengal, malachite green, cyanine dyes, methylene blue, and porphyrin dyes. Other photosensitizers are well known in the art and are widely used in photodynamic therapy (PDT) for cancer.

[0304] In some embodiments, the ROS source comprises titania (TiO2). In some embodiments, the titania is in the form of nano- or microparticles. In some embodiments, the titania particles have a diameter of 10 - 30 nm, 30 - 100 nm, 100 nm - 200 nm, 200 - 500 nm, 500 - 800 nm, 800 nm - 1 μm, 1 - 5 μm, 5 - 10 μm, 10 - 100 μm (including any range therebetween).

[0305] TiO2 particles generate electrons (e - ) and positively charged holes (h + ) in the conduction band and valence band of the semiconductor material, respectively, by absorption of light in the UV-A region. Then, in the presence of molecular oxygen, reactive oxygen species (ROS, O2 .- , HO2 . , H2O2 and mainly HO . ) are generated.

[0306] In some embodiments, the ROS source comprises a peroxide source. In some embodiments, the peroxide source is selected from the group consisting of hydrogen peroxide, urea hydrogen peroxide, metal peroxides (such as sodium peroxide, calcium peroxide, etc.) and / or their derivatives, percarbonates (such as sodium percarbonate, calcium percarbonate, etc.) and / or their derivatives, periodates (such as sodium periodate, etc.) and / or their derivatives, persulfates (such as sodium persulfate, ammonium persulfate, etc.) and / or their derivatives, perborates (such as sodium perborate, etc.) and / or their derivatives, silver(II) oxide, perbenzoic acid and / or its derivatives (such as chloroperbenzoic acid or their salts), perchloric acid or its salts, chlorine dioxide, benzoyl peroxide, ketone peroxide, peroxydicarbonate, peroxyester, dialkyl peroxide, peroxyacetic acid (PAA), hydroperoxide, peroxyketal or any combination thereof.

[0307] In some embodiments, the carboxylic acid includes short-chain carboxylic acids. In some embodiments, the carboxylic acid includes propionic acid, acetic acid, butyric acid and α-hydroxycarboxylic acids (e.g., lactic acid), citric acid, amino acids, or any combination thereof. In some embodiments, the oxidizing agent includes silver salts and hydrogen peroxide. In some embodiments, the oxidizing agent is a peroxycarboxylic acid. In some embodiments, the oxidizing agent includes hydrogen peroxide. In some embodiments, the oxidizing agent includes hypochlorite, chlorite, chlorate, perchlorate, hexavalent chromium compounds (chromic acid and dichromate and chromium trioxide, pyridinium chlorochromate (PCC), dichromate, etc.), permanganate compounds (potassium permanganate, etc.), sodium perborate, nitrous oxide (N2O), nitrogen dioxide, dinitrogen tetroxide (NO2 / N2O4), potassium nitrate (KNO3), sodium bismuthate, cerium(IV) compounds (cerium ammonium nitrate and cerium sulfate, etc.), or any combination thereof.

[0308] In some embodiments, the electrophilic reactive group includes any or any combination thereof of aldehydes, ketones, oximes, acyl halides, active esters (e.g., N-hydroxysuccinimide), chloroformates, anhydrides, benzaldehyde (CHO), epoxides, isocyanates (such as carbamate derivatives of hexamethylene diisocyanate), thiols (such as ester derivatives of thiopropionic acid), benzaldehyde (such as ester derivatives of 4-formylbenzoic acid), isothiocyanates, maleimides, carbonates, sulfonyl chlorides, haloacetamides, acyl azides, imide esters, carbodiimides, vinyl sulfones, thiols (SH), C1-C10 thioalkyls, orthopyridyl disulfides.

[0309] In some embodiments, the antimicrobial polymer includes cationic polymers (chitosan, polylysine, and polyethyleneimine), N-haramine polymers, N-haramid polymers, or any combination thereof. In some embodiments, the antimicrobial agent is an antimicrobial peptide (such as KLAKLAK, KALA, etc.).

[0310] In some embodiments, the antimicrobial agent includes salicylic acid, chlorhexidine, benzalkonium chloride, ethanol, glutaraldehyde, formaldehyde, hydrogen peroxide, and sodium hypochlorite.

[0311] In some embodiments, the antimicrobial agent is adsorbed onto the MaSp fibers. In some embodiments, the antimicrobial agent is in contact with the MaSp fibers to form a layer. In some embodiments, this layer is an outer layer. In some embodiments, this layer is in the form of a homogeneous layer. In some embodiments, the MaSp fibers are coated with the antimicrobial agent. In some embodiments, the MaSp fibers are modified with an antimicrobial agent, where the antimicrobial agent is as described herein. In some embodiments, the fiber comprises the MaSp fibers and a further polymer, where the further polymer is as described herein.

[0312] In some embodiments, the composition comprises 0.01% to 50%, 0.01% to 1%, 1% to 5%, 5% to 10%, 10% to 15%, 15% to 20%, and 20% to 50% (w / w) (including any range between them) of MaSp fibers and further polymers.

[0313] In another aspect of the present invention, an antimicrobial composition is provided. In some embodiments, the antimicrobial composition is configured to capture (immobilize) pathogens. In some embodiments, the antimicrobial composition is configured to prevent parasitism (e.g., bacterial or viral infection). In some embodiments, the antimicrobial composition is configured to prevent pathogens from moving into cells (e.g., human cells). In some embodiments, the antimicrobial composition is characterized by controlled release properties with respect to one or more antimicrobial agents (e.g., carboxylic acids) described herein.

[0314] In some embodiments, the antimicrobial composition comprising MaSp fibers and / or derivatized MaSp fibers of the present invention and an antimicrobial agent (such as glycolic acid and / or lactic acid) adsorbed thereon is characterized by a sustained-release profile of the antimicrobial agent (glycolic acid and / or lactic acid). Such a sustained-release profile has been demonstrated by the inventors.

[0315] In some embodiments, the compositions of the present invention are intended for use in imparting antimicrobial effects to any one of the following: creams, foams, beads, gels, sprays, films, nonwoven meshes, textiles, and substrates. In some embodiments, the compositions of the present invention are in the form of coatings.

[0316] In some embodiments, the composition of the present invention comprises a derivatized MaSp fiber, where the functional portion is a carboxylic acid or contains one. In some embodiments, the carboxylic acid has a polar, i.e., charged side chain, such as cysteic acid, thereby increasing the surface wettability of the fiber to form a superhydrophilic surface. In some embodiments, the surface of the MaSp fiber and / or the derivatized MaSp fiber of the present invention is at least partially modified with cysteic acid.

[0317] In some embodiments, the surface of the fiber is modified with covalently bonded cysteic acid and further bonded to antimicrobial silver particles. In some embodiments, the silver particles have diameters of 1–100 nm, 10–10 nm, 20–50 nm, and 50–100 nm (including any range in between). The combination of a superhydrophilic surface (e.g., having a water contact angle of less than 90°) and antimicrobial silver particles is claimed to be sufficient to induce the inactivation of captured microorganisms (e.g., viruses or bacteria).

[0318] In some embodiments, the composition of the present invention comprises the derivatized MaSp fiber of the present invention, where the functional portion is or comprises an electrophilic reactive group (aldehyde, ketone, oxime, acyl halogen, active ester (e.g., N-hydroxysuccinimide), chloroformate, anhydride, benzaldehyde, etc.). In some embodiments, the electrophilic reactive group induces crosslinking of pathogens (e.g., viruses and / or bacteria) captured by or in contact with the fiber of the present invention. In some embodiments, the electrophilic reactive group induces covalent bond formation with pathogens.

[0319] In some embodiments, the composition of the present invention comprises a derivatized MaSp-based fiber of the present invention, where the functional portion is a polymerizable monomer (e.g., lactone) and optionally a polymerization catalyst (e.g., a Sn-based catalyst). In some embodiments, the composition of the present invention induces polymerization of the monomer upon contact with a pathogen. Such polymerization may involve the formation of polymer chains (e.g., polyester such as PLA) in contact with the pathogen to capture the pathogen on or within the fiber composition of the present invention. In some embodiments, the in-situ polymerized chains capture and / or deactivate the pathogen.

[0320] In some embodiments, the composition of the present invention comprises a derivatized MaSp fiber of the present invention bonded to an antimicrobial polymer (such as polyglutaraldehyde). In some embodiments, the composition of the present invention comprises a derivatized MaSp fiber in contact with or bonded to an antimicrobial metal (such as copper, silver, zinc, nickel, cobalt, gold, or a combination thereof) or an antimicrobial metal salt.

[0321] In some embodiments, the composition is found in concentrations of 0.01%~80% (w / w), 0.05%~80% (w / w), 0.09%~80% (w / w), 0.1%~80% (w / w), 0.5%~80% (w / w), 0.9%~80% (w / w), 1%~80% (w / w), 5%~80% (w / w), 10%~80% (w / w), 15%~80% (w / w), 20%~80% (w / w), 30%~80% (w / w), and 50%~80% (w / w). Contains antimicrobial agents in amounts of w / w, 0.01%~50%(w / w), 0.05%~50%(w / w), 0.09%~50%(w / w), 0.1%~50%(w / w), 0.5%~50%(w / w), 0.9%~50%(w / w), 1%~50%(w / w), 5%~50%(w / w), 10%~50%(w / w), 15%~50%(w / w), 20%~50%(w / w), or 30%~50%(w / w) (including any range in between).

[0322] In some embodiments, the composition is found to be present in concentrations of 0.001%~95% (w / w), 0.005%~95% (w / w), 0.009%~95% (w / w), 0.01%~95% (w / w), 0.05%~95% (w / w), 0.09%~95% (w / w), 0.1%~95% (w / w), 0.5%~95% (w / w), 0.9%~95% (w / w), 1%~95% (w / w), and 5%~ 95%(w / w), 10%~95%(w / w), 15%~95%(w / w), 20%~95%(w / w), 30%~95%(w / w), 50%~95%(w / w), 0.01%~80%( w / w), 0.05%~80%(w / w), 0.09%~80%(w / w), 0.1%~80%(w / w), 0.5%~80%(w / w), 0.9%~80%(w / w), 1%~80%( w / w), 5%~80%(w / w), 10%~80%(w / w), 15%~80%(w / w), 20%~80%(w / w), 30%~80%(w / w), 50%~80%(w / w), 0 .001%~50%(w / w), 0.005%~50%(w / w), 0.009%~50%(w / w), 0.01%~95%(w / w), 0.01%~50%(w / w), 0.05%~5 Includes MaSp fibers in the following percentages: 0% (w / w), 0.09%~50% (w / w), 0.1%~50% (w / w), 0.5%~50% (w / w), 0.9%~50% (w / w), 1%~50% (w / w), 5%~50% (w / w), 10%~50% (w / w), 15%~50% (w / w), 20%~50% (w / w), or 30%~50% (w / w) (including any range in between).

[0323] In some embodiments, the ratio of MaSp fiber to antimicrobial agent is 0.01:1 to 1:1, 0.02:1 to 1:1, 0.05:1 to 1:1, 0.09:1 to 1:1, 0.1:1 to 1:1, 0.5:1 to 1:1, or 0.9:1 to 1:1 (including any range in between).

[0324] In some embodiments, the ratio of MaSp fiber to antibacterial agent is 100:1~1:100, 95:1~1:100, 80:1~1:100, 60:1~1:100, 50:1~1:100, 30:1~1:100, 20:1~1:100, 10:1~1:100, 9:1~1:100, 5:1~1:100, 2:1~1:100, 100:1~1:80, 95:1~1:80, 80:1~1:80, 60:1~1:80, 50:1~1:80, 30:1~1:80, 20:1~1:80, 10:1~1:80, 9:1~1:80, 5:1~1:80, The ranges are 2:1~1:80, 100:1~1:50, 95:1~1:50, 80:1~1:50, 60:1~1:50, 50:1~1:50, 30:1~1:50, 20:1~1:50, 10:1~1:50, 9:1~1:50, 5:1~1:50, 2:1~1:50, 100:1~1:10, 95:1~1:10, 80:1~1:10, 60:1~1:10, 50:1~1:10, 30:1~1:10, 20:1~1:10, 10:1~1:10, 9:1~1:10, 5:1~1:10, or 2:1~1:10 (including any range in between).

[0325] In another aspect of the present invention, a composition or article is provided comprising MaSp-based fibers in contact with or bonded (e.g., non-covalently) to a metal. In some embodiments, the composition comprises layered metal fibers, where the fibers comprise the MaSp-based fibers described herein and optionally further polymers. In some embodiments, the fibers are in contact with or coated with a polymer layer, where the polymer layer is in contact with or coated with a metal, where the metal is as described herein. In some embodiments, the fibers are at least partially coated with a polymer layer.

[0326] In some embodiments, the composition or article comprises a further polymer reinforced with MaSp fibers. In some embodiments, the reinforcement is 1-50%, 5-10%, 10-15%, 15-20% (including any range in between). In some embodiments, the further polymer reinforced with MaSp fibers is in the form of fibers or yarns. In some embodiments, the further polymer reinforced with MaSp fibers is in the form of layers. In some embodiments, the further polymer reinforced with MaSp fibers is in the form of a fibrous material.

[0327] In some embodiments, the composition or article comprising the fibers coated with a polymer layer is a multilayer composition, where the polymer layer is bonded to an outer layer containing a metal. In some embodiments, the polymer layer is homogeneous. In some embodiments, the polymer layer is in the form of a coating. In some embodiments, the polymer layer partially coats the fibers.

[0328] In some embodiments, the polymer (also referred to herein as the coating polymer) is bonded to or trapped within the fibrils or pores of the MaSp fiber. In some embodiments, the coating polymer is electrostatically bonded to the MaSp fiber. In some embodiments, the coating polymer is attached to the MaSp fiber and / or the fiber. In some embodiments, the coating polymer is attached to a further polymer containing the fiber.

[0329] In some embodiments, further polymers are selected from the group consisting of thermoplastic polymers, thermosetting polymers, epoxy, polyester, polyamide, polyol, polyurethane, polyethylene, nylon, polyacrylate, polycarbonate, polyaldehyde, polycarboxylic acid, polyamine, polyimine, polylactic acid (PLA) or its copolymers, silicon, liquid crystal polymer, maleic anhydride grafted polypropylene, polycaprolactone (PCL), rubber, cellulose, or any combination thereof.

[0330] In some embodiments, articles comprising compositions described herein are provided. In some embodiments, the articles are antimicrobial articles. In some embodiments, the articles are conductive devices. In some embodiments, the articles are conductive substrates. In some embodiments, the articles are in the form of coated glass substrates.

[0331] In some embodiments, the article is a cosmetic article comprising a derivatized (e.g., aminated or PEI-modified) MaSp-based fiber of the present invention having a positive zeta potential. In some embodiments, the article is a cosmetic article comprising a composite material of the present invention (e.g., metal oxide particles complexed with the derivatized MaSp-based fiber of the present invention). In some embodiments, the article is an electronic device. In some embodiments, the article is an antimicrobial coating. In some embodiments, the article is a woven or nonwoven fabric. In some embodiments, the article is a fiber or yarn reinforced with a metal-doped fiber of the present invention.

[0332] In some embodiments, a kit comprising the composition or composite material of the present invention is provided. In some embodiments, the kit further comprises further materials, such as polymers, glass, ceramics, particles (e.g., carbon particles), or metal substrates.

[0333] The thickness of the polymer layer is 1 nm to 1 μm, 1 to 50 nm, 1 to 10 nm, 10 to 50 nm, 50 to 100 nm, 100 to 300 nm, 300 to 500 nm, 500 to 1000 nm (including any range in between). In some embodiments, the polymer layer includes a coating polymer.

[0334] In some embodiments, the polymer layer comprises one or more polymers. In some embodiments, the coating polymer is compatible with MaSp-based fibers (e.g., having sufficient adhesive strength to MaSp-based fibers). In some embodiments, the coating polymer is compatible with further polymers within the fibers. In some embodiments, the coating polymer is compatible with metals. In some embodiments, the coating polymer is compatible with metals or the fibers of the present invention.

[0335] In some embodiments, the coating polymer contains polar atoms or polar groups. In some embodiments, the polar atoms or polar groups are intended to provide sufficient affinity (i.e., adhesive strength) to the metal (such as Pd). In some embodiments, the coating polymer contains atoms having an electronegativity less than that of carbon. In some embodiments, the coating polymer contains N, O, S, or a combination thereof. In some embodiments, the coating polymer contains amino, hydroxy, carbonyl, carboxy, ester, ether, amide, or a combination thereof.

[0336] In some embodiments, the coating polymer includes polyethyleneimine (PEI), polylysine, polyarginine, polyester (e.g., PLA, PCL), polyamide (nylon), polyvinyl alcohol, polyether (PEG), or a combination thereof. In some embodiments, the coating polymer includes PEI. In some embodiments, the coating polymer is bonded to a further polymer layer.

[0337] In some embodiments, the polymer layer is in contact with or bonded to the metal. In some embodiments, the metal is uniformly dispersed on the outer surface of the polymer layer. In some embodiments, the metal forms a layer on top of the polymer layer. In some embodiments, the metal layer is 1 to 10 atoms thick (including any range in between). In some embodiments, the metal is in colloidal form. In some embodiments, the metal is in the form of particles ranging from 1 to 500 nm.

[0338] In some embodiments, the polymer layer is in contact with or bonded to a first metal layer containing a first metal. In some embodiments, the polymer layer is in contact with or bonded to a first metal layer and a second metal layer, where the second metal layer contains a second metal. In some embodiments, the first metal and the second metal are the same. In some embodiments, the first metal and the second metal are different.

[0339] In some embodiments, a metal layer (e.g., a first metal layer) in contact with or bonded to a polymer layer forms multiple aggregation sites for a second metal. In some embodiments, the first metal has a high affinity for the coating polymer of the polymer layer. In some embodiments, the first metal promotes the deposition of the second metal (e.g., by electroless plating). In some embodiments, the first metal can reduce a salt of the second metal, thereby forming a second metal layer on or in contact with the first metal. In some embodiments, the first metal is bonded to a coating polymer.

[0340] In some embodiments, the second metal is bonded to or aggregated on the first metal. In some embodiments, the second metal forms a layer on top of the first metal layer. In some embodiments, the second metal has an affinity for the first metal. In some embodiments, the second and first metals are in the form of a layered structure, where each metal layer is separated. In some embodiments, the second and first metals are mixed together in a metal layer. In some embodiments, the second and first metals are mixed together to form a single metal layer on top of a polymer layer. In some embodiments, the fiber is at least partially coated with either the first and second metals or a combination of the first and second metals.

[0341] In some embodiments, the metal layer comprises a chemically reducible metal as the first metal, where chemically reducible means reduction of the first metal salt by a chemical reduction process. In some embodiments, the metal layer comprises an antimicrobial metal as the second metal.

[0342] In some embodiments, the metal layer does not contain a second metal. In some embodiments, the metal layer contains at least 90%, at least 95%, and at least 99% of the first metal, where the first metal is an antimicrobial metal. In some embodiments, the first metal and optionally the second metal are transition metals. Transition metals are well known in the art and are called d-electron metals.

[0343] In some embodiments, the antimicrobial metal includes copper, silver, zinc, or a combination thereof. In some embodiments, the first metal or a salt thereof has a reduction potential suitable for chemical reduction. In some embodiments, the first and second metals are suitable for electroless plating methods.

[0344] In some embodiments, the composition of the present invention comprises (i) a fiber comprising a MaSp system and optionally a further polymer (e.g., nylon or PCL), coated with (ii); a polymer layer comprising (ii) a coating polymer (e.g., PEI), in contact with or attached to (iii); and a metal layer comprising (iii) a first metal (e.g., Pd, Cu, Ag, Zn and optionally salts thereof), and further comprising a second metal such as an antimicrobial metal (e.g., Cu or Ag).

[0345] In some embodiments, the multilayer fiber compositions of the present invention are illustrated by Figure 1.

[0346] In some embodiments, compositions containing MaSp fibers or compositions containing MaSp fibers and a further polymer (e.g., nylon) promote the bonding of metals thereto. In some embodiments, compositions containing MaSp fibers or compositions containing MaSp fibers and a further polymer (e.g., nylon) are characterized by improved metal bonding affinity compared to polymers without MaSp fibers as illustrated by Figure 2. In some embodiments, the fibers or metal-coated fibers of the present invention are characterized by an antimicrobial effect. In some embodiments, the fibers or metal-coated fibers of the present invention are characterized by an improved antimicrobial effect compared to other coated polymers or uncoated polymers. In some embodiments, this antimicrobial effect is illustrated by Figure 4.

[0347] In some embodiments, the composition of the present invention is in the form of a coating on a substrate. In some embodiments, the composition of the present invention forms a mixture with a substrate. In some embodiments, the composition of the present invention is in the form of an antimicrobial additive (e.g., a mixture or a coating). In some embodiments, the composition (e.g., the fiber) is characterized by wettability. In some embodiments, the wettability is suitable for allowing contact between a droplet (e.g., containing a virus particle) and the droplet for a sufficient time to inactivate microorganisms within the droplet. In some embodiments, this wettability is 100 o Less than 90 o Less than 80 o Less than 70 o Less than 60 o It is characterized by a contact angle less than (including any value in between).

[0348] In some embodiments, this wettability is achieved by incorporating superhydrophobic particles or polar molecules (such as cysteic acid) onto or into the MaSp fibers. In some embodiments, the superhydrophobic particles include cysteic acid.

[0349] In some embodiments, the substrate:antimicrobial composition ratio is 1000:1~1:100, 1000:1~1:50, 1000:1~1:10, 1000:1~1:1, 900:1~1:100, 900:1~1:10, 900:1~1:1, 500:1~1:100, 500:1~1:10, 500:1~1:1, 100:1~1:100, 95:1~1:100, 80:1~1:100, 60:1~1:100, 50:1~1:100, 30:1~1:100, 20:1~1:100, 10:1~1:100, 9:1~1:100, 5:1~1:100, 2:1~1:100, 100:1~1:80, 95:1~1:80, 80:1~1:80, 60:1~1:80, 50 :1~1:80, 30:1~1:80, 20:1~1:80, 10:1~1:80, 9:1~1:80, 5:1~1:80, 2:1~1:80, 100:1~1:50, 95:1~1:50, 80:1~1:50, 60:1~1:50, 50:1~1:50, 30:1~1:50, 20:1~1:50, 10:1~1:50, 9:1~ These ranges are 1:50, 5:1~1:50, 2:1~1:50, 100:1~1:10, 95:1~1:10, 80:1~1:10, 60:1~1:10, 50:1~1:10, 30:1~1:10, 20:1~1:10, 10:1~1:10, 9:1~1:10, 5:1~1:10, or 2:1~1:10 (including any range in between).

[0350] In some embodiments, the substrate includes one of the following: a textile substrate, a fabric, a polymer substrate, a glass substrate, and a metal substrate.

[0351] In another aspect of the present invention, a composition is provided comprising the fibers of the present invention covalently bonded to an antimicrobial agent, wherein the antimicrobial agent is as described herein.

[0352] In some embodiments, the fiber, or MaSp fiber, is covalently bonded to the antimicrobial agent via a linker. In some embodiments, the linker is bonded to the tyrosine of the MaSp fiber. In some embodiments, the linker is bonded to the tyrosine via a diazo group. In some embodiments, the phenol ring of tyrosine undergoes diazotization to form a covalent bond with the linker, thereby forming the antimicrobial agent covalently bonded to the MaSp fiber.

[0353] In some embodiments, the MaSp fibers bound to the antimicrobial agent are as follows: [ka] As shown in the formula, where R contains an antimicrobial agent as described herein, [ka] R represents a MaSp-based fiber. In some embodiments, R includes a polymer or antimicrobial peptide such as an antimicrobial polymer (e.g., PGA) as described herein.

[0354] In some embodiments, the linker is 2-(4-aminophenyl)ethylamine. In some embodiments, the MaSp fiber is bonded to the antimicrobial agent via diazotized 2-(4-aminophenyl)ethylamine. In some embodiments, R containing the antimicrobial agent is as follows: [ka] Represented by one of the structures provided in the formula, where X represents a heteroatom, n is an integer from 0 to 10, the dye contains a photosensitizer, and R includes an optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, or a combination thereof.

[0355] General matters As used herein, the term "approximately" refers to a range of ±10%.

[0356] The terms "comprises," "comprising," "includes," "including," and "having," as well as their conjugations, all mean "to include, but not to limit."

[0357] The phrase "consisting of" means "including and limited to."

[0358] The phrase "consisting essentially of" means that the composition, method, or structure may include further components, steps, and / or parts, but only if the further components, steps, and / or parts do not substantially alter the basic and novel properties of the claimed composition, method, or structure.

[0359] As used herein, the term “stablely encapsulated” refers to the ability of the composition to substantially prevent the release of the active ingredient therefrom. As used herein, the term “substantially prevent” refers to the total amount of active ingredient removed by the first tape strip and the second tape strip, as measured by a skin tape test (described in the Examples section).

[0360] In some embodiments, the materials substantially include at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, and at least 99% (including any value in between). As used herein, the term “bonded” refers to covalent bonds (e.g., coordinate bonds, single bonds, double bonds, triple bonds, etc.), non-covalent bonds, physical interactions, or combinations thereof. Non-covalent bonds are well known in the art, and include, in particular, hydrogen bonds, pp-stacking, and van der Waals interactions.

[0361] The term “exemplary” is used herein to mean “to serve as an example, illustration, or representation.” Any embodiment described as “exemplary” should not necessarily be construed as being preferable or advantageous to other embodiments, and / or as precluding the incorporation of features from other embodiments.

[0362] The term “optional” is used herein to mean “provided in some embodiments and not provided in other embodiments.” Any particular embodiment of the present invention may include multiple “optional” features, as long as such features do not contradict each other.

[0363] As used herein, the singular forms “a,” “an,” and “the” refer to multiple subjects unless the context clearly indicates otherwise. For example, the terms “compound” or “at least one compound” may refer to multiple compounds, including mixtures thereof.

[0364] definition As used herein, the term "alkyl" refers to aliphatic hydrocarbons containing linear and branched groups. Preferably, alkyl groups have 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms (or C1-C6 alkyl). Thus, short alkyl groups have 20 or fewer main chain carbon atoms. Alkyl groups may be substituted or unsubstituted as defined herein.

[0365] As used herein, the term “alkyl” includes saturated or unsaturated hydrocarbons, and therefore further includes alkenyls and alkynyls. As used herein, the term “C1-C6 alkyl” includes any C1-C6 alkyl-related compound and refers to any linear or branched alkyl chain containing 1-6, 1-2, 2-3, 3-4, 4-5, or 5-6 carbon atoms (including any range in between). In some embodiments, the C1-C6 alkyl includes any or any combination thereof of methyl, ethyl, propyl, butyl, pentyl, isopentyl, hexyl, and tert-butyl. In some embodiments, the C1-C6 alkyl described herein further includes an unsaturated bond, where the unsaturated bond is located at position 1, 2, 3, 4, 5, or 6 of the C1-C6 alkyl.

[0366] The term "C1-C6 alkyl-related compounds" used herein includes any C1-C6 alkyl-related compounds. 10 The term "alkyl" refers to any linear or branched alkyl chain containing 1 to 6, 1 to 2, 2 to 3, 3 to 4, 4 to 5, 5 to 6, 6 to 8, or 8 to 10 carbon atoms (including any range in between). In some embodiments, C1 to C 10 Alkyl includes any or any combination thereof of methyl, ethyl, propyl, butyl, pentyl, isopentyl, hexyl, nonyl, decyl, and tert-butyl. In some embodiments, C1-C described herein 10 Alkyl further contains unsaturated bonds, where the unsaturated bonds are C1-C 10 It is located at the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, or 10th position of the alkyl group.

[0367] The term "alkenyl" refers to an unsaturated alkyl group as defined herein, having at least two carbon atoms and at least one carbon-carbon double bond. Alkenyls may be substituted or unsubstituted by one or more substituents described herein.

[0368] The term "alkynyl" as defined herein refers to an unsaturated alkyl having at least two carbon atoms and at least one carbon-carbon triple bond. The alkynyl may be substituted or unsubstituted by one or more substituents described herein.

[0369] The term "cycloalkyl" describes any monocyclic or fused ring (i.e., a ring sharing adjacent pairs of carbon atoms) group in which one or more of the rings do not have a fully conjugated π-electron system. Cycloalkyl groups may be substituted or unsubstituted as shown herein. In some embodiments, the term "cycloalkyl" refers to a C3-C10 cyclic ring. In some embodiments, the term "cycloalkyl" refers to a C3-C10 cyclic ring containing one, two, three, or four heteroatoms (e.g., N, NH, O, or S). (C3-C 10 The ) ring refers to an optionally substituted C3, C4, C5, C6, C7, C8, C9, or C10 ring. In some embodiments, (C3~C 10 The ring contains optionally substituted cyclopropane, cyclobutene, cyclopentane, cyclohexane, or cycloheptane.

[0370] The term "aryl" refers to all carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) groups that have a fully conjugated π-electron system. The term "aryl" refers to aromatic (C6~C) groups. 12 ) represents a ring. The aryl group may be substituted or unsubstituted as shown herein. The term "(C6~C)" used herein refers to a ring. 12 The term "(C6~C11 or C12 aromatic ring)" refers to any substituted C6, C7, C8, C9, C10, C11 or C12 aromatic ring. In some embodiments, (C6~C11 or C12 12 Aromatic rings refer to bicyclic aryl or bicyclic heteroaryl rings (e.g., fused rings, spiro rings, and biaryl rings).

[0371] The term "bicyclic heteroaryl" as used herein refers to (C6-C6) 12) refers to a bicyclic heteroaryl ring, and here it refers to a bicyclic (C6~C 10 The ring is as described herein.

[0372] As used herein, the term “bicyclic aryl” means (C6~C 12 ) refers to a bicyclic aryl ring, and here it refers to a bicyclic (C6~C 12 The ring is as described herein.

[0373] The term "alkoxy" refers to both O-alkyl and -O-cycloalkyl groups as defined herein.

[0374] The term "aryloxy" represents the -O-aryl as defined herein.

[0375] In the general formulas herein, the alkyl, cycloalkyl, and aryl groups may each be substituted with one or more substituents, where each substituent independently may be a halide, alkyl, alkoxy, cycloalkyl, nitro, amino, hydroxyl, thiol, thioalkoxy, carboxy, amide, aryl, and aryloxy, depending on the substituted group and its position in the molecule. Further substituents are also possible.

[0376] The terms "halogen," "halogen," or "halo" refer to fluorine, chlorine, bromine, or iodine.

[0377] The term "haloalkyl" refers to an alkyl group as defined herein that is further substituted with one or more halides.

[0378] The term "haloalkoxy" refers to an alkoxy group as defined herein that is further substituted with one or more halides.

[0379] The term "hydroxyl" or "hydroxy" refers to the -OH group.

[0380] The terms "mercapto" or "thiol" represent the -SH group.

[0381] The term "thioalkoxy" refers to both -S-alkyl groups and -S-cycloalkyl groups as defined herein.

[0382] The term "thioaryloxy" refers to both -S-aryl and -S-heteroaryl groups as defined herein.

[0383] The term "amino" describes the -NR'R'' group, where R' and R'' are as defined herein.

[0384] The term "heterocyclyl" refers to a monocyclic or fused ring group having one or more atoms, such as nitrogen, oxygen, and sulfur, in the ring. The ring may also have one or more double bonds. However, the ring does not have a fully conjugated π-electron system. Typical examples include piperidine, piperazine, tetrahydrofuran, tetrahydropyran, and morpholino.

[0385] The terms "carboxy" or "carboxylate" describe a -C(O)OR' group, where R' is hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl (linked by a ring carbon), or heterocyclyl (linked by a ring carbon) as defined herein.

[0386] The term "carbonyl" describes the -C(O)R' group, where R' is as defined herein.

[0387] The above terms also include their thio derivatives (thiocarboxy and thiocarbonyl).

[0388] The term "thiocarbonyl" describes the -C(S)R' group, where R' is as defined herein.

[0389] The "thiocarboxy" group describes the -C(S)OR' group, where R' is as defined herein.

[0390] The "sulfinyl" group describes the -S(O)R' group, where R' is as defined herein.

[0391] The "sulfonyl" or "sulfonate" group describes the -S(O)2R' group, where R' is as defined herein.

[0392] The term "carbamyl" or "carbamate group" describes the -OC(O)NR'R'' group, where R' is as defined herein and R'' is as defined for R'.

[0393] The "nitro" group refers to the -NO2 group.

[0394] As used herein, the term "amide" encompasses C-amides and N-amides.

[0395] The term "C-amide" describes a -C(O)NR'R'' terminal group or -C(O)NR'- bonded group, which are defined herein, where R' and R'' are as defined herein.

[0396] The term "N-amide" describes an -NR''C(O)R' terminal group or an -NR'C(O)- bonded group, which are defined herein, where R' and R'' are as defined herein.

[0397] As used herein, the term “carboxylic acid derivative” includes carboxylates, amides, carbonyls, anhydrides, carbonates, and carbamates.

[0398] The "cyano" or "nitrile" group refers to the -CN group.

[0399] The terms "azo" or "diazo" describe an -N=NR' terminal group or -N=N- bonded group, which are defined herein, and in formulas, R' is as defined herein.

[0400] The term "guanidine" describes the -R'NC(N)NR''R''' terminal group or the -R'NC(N)NR''- bonded group, which are defined herein, wherein R', R'' and R''' are as defined herein.

[0401] As used herein, the term "azide" refers to the -N3 group.

[0402] The term "sulfonamide" refers to the -S(O)2NR'R'' group, where R' and R'' are as defined herein.

[0403] The terms "phosphonyl" or "phosphonic acid ester" describe the -OP(O)-(OR')2 group, where R' is as defined herein.

[0404] The term "phosphenyl" describes the -PR'R'' group, where R' and R'' are as defined herein.

[0405] The term "alkylaryl" refers to an alkyl group as defined herein, substituted with an aryl group as described herein. An exemplary alkylaryl is benzyl.

[0406] The term "heteroaryl" refers to a monocyclic (e.g., C5-C6 heteroaryl ring) or fused ring (i.e., a ring sharing adjacent pairs of atoms) group having one or more atoms in the ring, such as nitrogen, oxygen, and sulfur, and further having a fully conjugated π-electron system. In some embodiments, the terms "heteroaryl" and "C5-C6 heteroaryl" are used interchangeably herein. Examples of heteroaryl groups, but not limited to, include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline, and purine. Heteroaryl groups may or may not be substituted with one or more substituents listed above herein. Typical examples include thiadiazole, pyridine, pyrrole, oxazole, indole, and purine.

[0407] As used herein, the terms “halo” and “halogenated” are interchangeable, but refer to the halogenated atoms that are fluorine, chlorine, bromine, or iodine, also known as fluorides, chlorides, bromides, and iodides.

[0408] The term "haloalkyl" refers to an alkyl group that is further substituted with one or more halides and defined on top of that.

[0409] The terms "substituted" or "substituent" refer to one, two, three, four, or five substituents, where each substituent independently includes (C0-C6) alkyl-aryl, (C0-C6) alkyl-heteroaryl, (C0-C6) alkyl-(C3-C8) cycloalkyl, optionally substituted C3-C8 heterocyclyl, halogen, -NO2, -CN, -OH, -CONH2, -CONR2, -CNNR2, -CSNR2, -CONH-OH, -CONH-NH2, -NHCOR, -NHCSR, -NHCNR, -NC(=O)OR, -NC(=O)NR, -NC(=S)OR, -NC(=S)NR, -SO2R, -SOR, -SR, -SO2OR, -SO2N(R)2, -NHNR2, - Selected from NNR, C1-C6 haloalkyl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(C1-C6 alkyl), hydroxy(C1-C6 alkoxy), alkoxy(C1-C6 alkyl), alkoxy(C1-C6 alkoxy), C1-C6 alkyl-NR2, C1-C6 alkyl-SR, -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -CO2H, -CO2R, -OCOR, -OCOR, -OC(=O)OR, -OC(=O)NR, -OC(=S)OR, -OC(=S)NR (including any combination thereof).

[0410] Throughout this application, various embodiments of the invention may be presented in range form. Naturally, range form is merely for convenience and brevity and should not be interpreted as an unchangeable limitation on the scope of the invention. Therefore, range descriptions should be interpreted to include all possible sub-ranges specifically disclosed, as well as the individual numbers within those ranges. For example, a range description such as 1-6 should be interpreted to include specifically disclosed sub-ranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, as well as the individual numbers within those ranges, e.g., 1, 2, 3, 4, 5, and 6. This applies regardless of the width of the range.

[0411] Wherever a range of numbers is indicated herein, it is intended to include any enumerated numbers (fractions or integers) within that range. The phrases “the range between the first and second indicators” and “the range between the first and second indicators” are used interchangeably herein and are intended to include the first and second indicators and all fractions and integers between them.

[0412] As used herein, the term “method” refers to a manner, means, techniques and procedures for accomplishing a given task, including, but not limited to, modes, means, techniques and procedures readily developed from methods, means, techniques and procedures known or publicly known by practitioners of the chemical, pharmacological, biological, biochemical and medical technology fields.

[0413] As used herein, the term “treat” includes inhibiting, substantially blocking, slowing or reversing the progression of a disease, substantially improving the clinical or aesthetic symptoms of a disease, or substantially preventing the appearance of the clinical or aesthetic symptoms of a disease.

[0414] For clarity, it should be understood that certain features of the Invention described in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, various features of the Invention described in the context of a single embodiment may also be provided separately or as any preferred partial or combination in any other described embodiment of the Invention. Certain features described in the context of different embodiments should not be considered essential features of those embodiments unless the embodiment cannot be carried out without those elements.

[0415] Various embodiments and aspects of the present invention, detailed herein and claimed in the following claims, are experimentally supported in the following examples. [Examples]

[0416] Next, the following embodiments illustrate some embodiments of the present invention in conjunction with the above description, in a non-limiting manner.

[0417] Example 1: Derivatized MaSp-based fiber The amination of MaSp fibers was performed as follows.

[0418] An aqueous dispersion containing MaSp fibers together with an anionic surfactant (also referred to herein as "SVX") was centrifuged and redispersed in deionized (DI) water. The MaSp fibers were then substantially dried according to a well-established procedure and redispersed in DI water to obtain a MaSp suspension.

[0419] An aqueous solution of 2-(4-aminophenyl)ethylamine (APEA) (1-20% w / w) was prepared and then acidified with HCl (1M).

[0420] An excess of 2 to 10 moles of NaNO2 was added to a 2-(4-aminophenyl)ethylamine solution, and the temperature was maintained at 1 to 10°C for 10 to 60 minutes to obtain a diazotized solution. The diazotized solution was then added dropwise to a MaSp suspension while cooling. The resulting mixture was stirred at the completion of the reaction.

[0421] Next, the supernatant was discarded, and the remaining derivatized MaSp fibers were thoroughly washed with DI water, after which the amination fibers were dried.

[0422] The modification efficiency (yield) was determined by calculating the amount of unreacted APEA in the reaction mixture. This calculation was performed by determining the UV absorption of APEA (by UV spectrophotometric method). A maximum of 60% tyrosine diazotization was observed.

[0423] The bonding of amination-modified MaSp fibers to polyglutaraldehyde (PGA) was carried out as follows.

[0424] Preparation of polyglutaraldehyde (average MW of 500-2000 or approximately 1000 Da): 20 mL of glutaraldehyde aqueous solution (25%) was added to a 1 M aqueous solution of K2CO3 and heated to 50°C for 2 hours. The mixture was then cooled to room temperature (20-25°C) and the pH was set to 7 using HCl (37%). The mixture was then centrifuged at 7000 rpm for 10 minutes, and the mixed solution was diluted with acetone to 10 times its volume. The K2CO3 powder was filtered, and the acetone was evaporated. The PGA aqueous solution was freeze-dried and maintained at -20°C.

[0425] Amination of MaSp fibers was added to 1-10 mL of polyglutaraldehyde (PGA) solution [0.01-2 M concentration] in aqueous buffer. The resulting suspension was then cooled to RT and the pH was neutralized with HCl (37%). This reaction was maintained overnight at 1-10°C with stirring to obtain the desired product: SVX-PGA.

[0426] The metal chelate group (IDA) was bonded to polyglutaraldehyde (PGA) as follows.

[0427] MaSp fibers derivatized with PGA were added to a 0.28 M solution of sodium iminodiacetate dibasic hydrate (IDA) buffered with HEPES (0.05 M), left overnight in an ice bath (1-10°C), and then washed by redispersion in HEPES buffer.

[0428] The reaction mixture was centrifuged, the precipitant was redispersed in 10 mL of aqueous buffer, then centrifuged again, and thoroughly washed with DI water. The resulting conjugate (PGA-IDA modified MaSp fiber) was freeze-dried.

[0429] Bonding of amination-modified MaSp fibers with poly(acrylic acid) (PAA) 1.84 g of poly(acrylic acid) (Aldrich 306223, average MW of approximately 3,000,000 Da) was suspended in 0.1 M MES (N-morpholino) ethanesulfonic acid) buffer solution overnight at RT with stirring. Next, amination of MaSp fibers (prepared as described above) was suspended in MES (0.1 M), and the suspension was added to the poly(acrylic acid) suspension. The mixture was left to stand with stirring for at least 4 hours until a homogeneous suspension was obtained. Next, 10-30 mg of EDC (1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide*HCl) aqueous solution was added to the suspension in 5 ml of DI water. Next, 40 mg of NHS (N-hydroxysulfosuccinimide sodium salt) aqueous solution was added to 5 ml of DI water. The resulting mixture was stirred overnight at RT (room temperature). The reaction mixture was centrifuged and resuspended in water several times. The resulting compound (PAA-modified MaSp fiber) was freeze-dried.

[0430] The molar or w / w ratio of PAA / MaSp fibers is 1695 cm². -1 Peak at (corresponding to PAA) and 1620cm -1 The peak intensity ratio of the peaks (corresponding to MaSp fibers) can be determined by FTIR spectroscopy. The peak intensity is normalized with respect to concentration based on a calibration curve.

[0431] The inventors have found a w / w ratio of PAA to MaSp fiber in the range of 1-200%, and 5*10 -4 ~5*10 -2 We successfully synthesized PAA-modified MaSp fibers using a molar ratio of PAA to MaSp fibers within a specified range.

[0432] Bonding of carboxylated MaSp fibers with poly(acrylamide) (PAAm) The carboxylation of MaSp fibers was carried out as follows.

[0433] An aqueous dispersion containing MaSp fibers together with an anionic surfactant (also referred to herein as "SVX") was centrifuged and redispersed in deionized (DI) water. The MaSp fibers were then substantially dried according to a well-established procedure and redispersed in DI water to obtain a MaSp suspension.

[0434] Aqueous solutions (1-20% w / w) of 4-aminobenzoic acid or 4-aminophenylacetic acid were prepared and then acidified with HCl (1M).

[0435] An excess of 2-10 moles of NaNO2 was added to a solution of 4-aminobenzoic acid or 4-aminophenylacetic acid, and the temperature was maintained at 1-10°C for 10-60 minutes to obtain a diazotized solution. The diazotized solution was then added dropwise to a MaSp suspension while cooling. The resulting mixture was stirred at the completion of the reaction.

[0436] Next, the supernatant was discarded, and the remaining derivatized MaSp fibers were thoroughly washed with DI water. Then, the carboxylated fibers were dried to obtain the desired product (SVX-COOH).

[0437] The modification efficiency (yield) was determined by calculating the amount of unreacted 4-aminobenzoic acid or 4-aminophenylacetic acid in the reaction mixture. This calculation was performed by determining the UV absorption of 4-aminobenzoic acid or 4-aminophenylacetic acid (by UV spectrophotometric method). A maximum of 60% tyrosine diazotization was observed.

[0438] Bonding of SVX-COOH and poly(acrylamide) (PAAm) by in situ polymerization.

[0439] 500 mg of acid-modified SVX-COOH (prepared as described above) was centrifuged in an aqueous suspension and resuspended in MES buffer (0.1 M).

[0440] 40 μL of DMPA3-(dimethylamino)-1-propylamine was added to the suspension. Then, 13.3 mg of an aqueous solution of EDC (1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide*HCl) was added to 5 ml of DI water, and then 20 mg of NHS (N-hydroxysulfosuccinimide sodium salt) was added to 5 ml of DI water. The reaction mixture was stirred in RT for 0.5 to 10 hours, then centrifuged, and washed with DI water.

[0441] Subsequently, 0.1-5 g of acrylamide was dissolved in 1-10 mL of water and added to the SVX-COOH-DMPA suspension, and the mixture was shaken vigorously. Then, 100 μL of 10% ammonium persulfate solution was added to the SVX-COOH-DMPA suspension, and the resulting mixture was stirred overnight at RT, then centrifuged, washed with DI water, and the desired conjugate was obtained by in-situ polymerization of PAAm on SVX-COOH.

[0442] The inventors have succeeded in synthesizing conjugates of derivatized MaSp fibers with various polymers such as PGA, PAA, PVA, PEI, PAAm, or combinations thereof (e.g., PGA-co-PEI). Furthermore, the inventors have succeeded in synthesizing the above-mentioned conjugates using MaSp proteins having mutated amino acid sequences (referred to herein as "mutated MaSp proteins").

[0443] Both MaSp proteins and mutated MaSp proteins exhibit a highly porous structure, resulting in very high porosity (e.g., at least 10 m) of derivatized MaSp fibers. 2 It is claimed that the BET surface area (determined by the BET surface area per g) is sequence-independent.

[0444] Example 2: Metal-doped derivatized MaSp-based fibers The inventors have successfully synthesized amination-modified MaSp fibers bonded to PGA, further bonding PGA to a metal chelating group (iminodiacetic acid or IDA=HN(CH2CO2H)2) ​​and synthesizing them according to the procedure of Example 1. Furthermore, they have succeeded in doping the PGA-IDA-modified MaSp fibers with Cu(0) deposited by an electroless plating method.

[0445] Pd doping of PGA-IDA-modified MaSp fibers: PGA-IDA-modified MaSp fibers were dispersed in 0.05 M HEPES buffer. Then, 3 mM aqueous palladium(II) acetate solution was added to the dispersion, and the reaction was maintained overnight at 1–10°C with stirring. The modified MaSp fibers were then separated by centrifugation and washed extensively with DI water. Subsequently, another portion of palladium(II) acetate was added to the modified MaSp fibers, and the reaction mixture was stirred at RT for 1–10 hours.

[0446] The reaction mixture was centrifuged, the precipitant was redispersed in 10 mL of aqueous buffer, then centrifuged again, and thoroughly washed with DI water. The resulting composite material (Pd-modified MaSp-based fiber) was freeze-dried.

[0447] The reduction of Pd to Pd(0): A reduction mixture based on lactic acid and boron was added to Pd-modified MaSp fibers to induce Pd(II) reduction and achieve Pd(0). The resulting gray fibers were washed several times with water to remove all traces of the unreacted reduction mixture.

[0448] Copper deposition: A suspension of Pd(0)-modified MaSp fibers was treated with a standard Cu solution for electroless plating. During this process, the blue color of Cu(II) disappeared, and the fibers turned brown. The resulting Cu(0)-doped MaSp fibers were washed several times with water.

[0449] Alternatively, copper colloids absorbed (non-covalently bonded) on or inside MaSp fibers (as shown in Figure 1) were prepared as follows.

[0450] Processing by PEI The original MaSp fibers, dispersed in water, were mixed with PEI at 40°C for 4 hours while stirring. The resulting suspension was washed several times with water and dried, resulting in the formation of an SVX-PEI composite material.

[0451] Nucleation by Pd: Sodium tetrachloropalladate (Na2PdCl4) was dissolved in water and added to the SVX-PEI composite material dispersed in water. This mixture was stirred for 2 hours to obtain yellow fibers. The resulting suspension of SVX-PEI-Pd(II) was washed with water to remove excess unreacted Pd salt.

[0452] Subsequently, reduction of Pd to Pd(0) and copper deposition were carried out as described above to obtain Cu colloid adsorbed onto the SVX-PEI composite. Then, nylon yarn reinforced with 10% Cu-doped fibers was prepared and tested for conductivity and antimicrobial activity.

[0453] As shown in Figure 4, nylon yarn reinforced with 10% Cu-doped fibers substantially prevented bacterial adhesion.

[0454] The conductivity of nylon yarn reinforced with 10% Cu(0) doped MaSp fibers was determined, and a highly conductive yarn (R=0.06Ω / m) was obtained. In contrast, derivatized MaSp fibers without metal doping showed 10 10 It is characterized by a resistivity exceeding Ω / m.

[0455] As shown in Figures 2 and 3, we successfully coated modified MaSp fibers and nylon fibers reinforced with them with palladium and then copper. In contrast, by applying the above procedure to "original" (i.e., unreinforced) nylon fibers, no Cu deposition was obtained on the original fibers. While not bound by any particular theory, it has been claimed that modified MaSp fibers promote metal (e.g., Pd and / or Cu) deposition.

[0456] Example 3: Hair coloring composition The inventors have successfully incorporated various derivatized porous MaSp fibers into hair coloring compositions (such as the hair coloring compositions described herein). Some of these hair coloring compositions resulted in a uniform and stable hair coating when applied to hair (e.g., human hair). Exemplary hair coloring compositions that successfully achieved hair coating include amination of MaSp fibers (e.g., chemically modified with 4-(2-aminoethyl)aniline, 3-aminopropyltriethoxysilane, or PEI). Furthermore, several dyes (cationic and anionic dyes) were used in the hair coating to obtain colored hair. The colored hair formed by applying the hair coloring compositions described herein retained its color even after extensive washing.

[0457] Furthermore, the inventors utilized various MaSp-based fibers for hair coloring compositions. Stable hair coatings were obtained by using amination-modified MaSp-based proteins and amination-modified mutant MaSp-based proteins.

[0458] Example 4: Sunscreen composition The inventors have succeeded in synthesizing MaSp-based fibers derivatized with PGA modified with a metal oxide chelating agent (succinic acid) represented by formula 4.

[0459] The modified MaSp fiber of formula 4 (SVX-PGA-SA) was synthesized by reacting SVX-PGA (prepared according to the procedure of Example 1) with 4-aminosalicylic acid as follows.

[0460] A 300 mg aqueous dispersion of SVX-PGA at pH 10.5 was mixed with 150 mg (excess) of 4-aminosalicylic acid at 20-40°C for 1-10 hours. The color of the reaction mixture changed to orange-yellow. The reaction mixture was then centrifuged, and the solid SVX-PGA-SA was washed several times to remove any unreacted aminosalicylic acid.

[0461] TiO2-SVX-PGA-SA composite material was obtained by complexing titania particles (particle size 300-500 nm) with SVX-PGA-SA as follows.

[0462] A 200 mg aqueous dispersion of SVX-PGA-SA at pH 3.5 was mixed with 200 mg of titanium dioxide at 40°C for 10 hours. The color of the reaction mixture turned crimson. The reaction mixture was then centrifuged, and the solid TiO2-SVX-PGA-SA was washed several times to remove any unreacted material.

[0463] The composite material of the present invention, containing titanium dioxide particles having a particle size greater than 300 nm, exhibited significantly improved dispersibility in aqueous and / or organic solutions compared to the original titania particles. An aqueous dispersion (e.g., represented by Formula 4) containing titanium dioxide particles (complexed) bonded to PGA-derivativeized MaSp fibers via salicylic acid showed superior stability compared to a control dispersion containing underivativeized MaSp fibers. Furthermore, the composite material containing titanium dioxide particles bonded to salicylic acid-derivativeized MaSp fibers exhibited excellent dispersibility (e.g., it can form a stable dispersion), where the w / w ratio of titanium dioxide particles to derivatized MaSp fibers is approximately 1:1.

[0464] The aforementioned composite material was used to reduce UV exposure (e.g., to human skin).

[0465] Example 5: Thermal stability of large bottle-shaped glandular spidoin protein (MaSp) polymers Analysis of differential scanning calorimetry (DSC) curves of the provided spider silk polymer (SVXE) expressed in bacteria reveals that SVXE does not exhibit a melting peak. Instead, it has a small glass transition temperature (T) of approximately 220°C and 280°C. g The region was shown. We were also able to observe a decomposition peak at approximately 330°C (Figure 8).

[0466] Figure 8 shows the DSC curves of the SVXE as the temperature rises from 25°C to 280°C (curve 1), cools to 50°C (curve 2), and rises again to 350°C (curve 3).

[0467] The thermogravimetric analysis (TGA) curve of SVXE shows that at a heating rate of 10°C / min, weight loss below 100°C was only about 5% of the absorbed water. A significant weight loss (over 1% / hour) can be observed starting above 230°C.

[0468] While the present invention has been described along with its specific embodiments, it is evident that many alternatives, modifications, and variations will be apparent to those skilled in the art. Therefore, it is intended to encompass all such alternatives, modifications, and variations that fall within the spirit and broad scope of the appended claims.

[0469] All publications, patents, and patent applications referenced herein are incorporated herein by reference to the same extent as each individual publication, patent, or patent application is specifically and individually indicated as being incorporated herein by reference. Furthermore, any citation or specification of references in this application should not be construed as an admission that such references are available to the present invention as prior art. Section headings, insofar as they are used, should not necessarily be construed as restrictive.

Claims

1. Derivatized porous bottle-shaped glandular spidoin protein (MaSp) fiber, The derivatized porous MaSp fiber is at least 10 m long. ２ Characterized by a BET surface area of ​​ / g, The derivatized porous MaSp fiber comprises (i) a functional moiety or (ii) a polymer covalently bonded to the tyrosine side chain of the porous MaSp fiber. The functionalized portion comprises one of the following: amino, carboxy, nitro, sulfonate, carbonyl, ester, anhydride, carbonate ester, carbamate, cyano, hydroxy, or any combination thereof. The functionalized portion or polymer is covalently bonded to the tyrosine side chain via a diazo bond, a silyl group, or any combination thereof. Derivatized porous MaSp-based fibers.

2. The derivatized porous MaSp fiber according to claim 1, wherein the load of the functionalized portion into the derivatized porous MaSp fiber is 0.01 μmol / g to 10 mmol / g.

3. Formula I: 【Chemistry 1】 It is expressed by, in the formula, 【Chemistry 2】 The derivatized porous MaSp fiber according to claim 1 or 2, wherein represents a porous MaSp fiber, A is selected from substituted or unsubstituted aryl, heteroaryl, and alkyl, R is the functionalized moiety or polymer, and the wavy bond represents a linker.

4. The derivatized porous MaSp fiber according to claim 3, wherein R is the polymer, and the linker comprises optionally substituted alkyl, amino acid, alkoxy, thioalkyl, aminoalkyl, glycol, -C(O)NH-, -C(O)O-, -C(O)-, -C(O)S-, -C(NH)NH-, -C(NH)O-, -C(NH)S-, urea, or any combination thereof.

5. The derivatized porous MaSp fiber according to claim 1, wherein the polymer is covalently bonded to a chelating agent, an antimicrobial agent, or any combination thereof, and the chelating agent comprises (i) a metal chelating group that can be bonded to a metal or a salt thereof, (ii) a metal oxide chelating group, or both (i) and (ii).

6. The derivatized porous MaSp fiber according to claim 5, wherein the metal chelating group comprises iminodiacetic acid (IDA), DOTA, NOTA, NODA, EDTA, HBED-CC (including any salts, derivatives, or combinations thereof).

7. The derivatized porous MaSp fiber according to claim 5, wherein the metal oxide chelating group is selected from salicylic acid, phosphonic acid, hydroxamic acid, malonic acid, pyrogallol, and 5-hydroxy-1,4-naphthoquinone (including any salt or any combination thereof).

8. The derivative porous MaSp fiber according to claim 1, wherein the polymer is selected from the group consisting of polyglutaraldehyde (PGA), polyvinyl alcohol (PVA), polyacrylic acid (PAA), polyethyleneimine (PEI), polyacrylamide (PAAm), polylysine, polyaniline, polyurethane, polyamide, polyvinyl chloride, silicone crosspolymer, polyvinylpyrrolidone, or any combination thereof.

9. The derivatized porous MaSp fiber according to claim 1, wherein the w / w ratio of the polymer to the porous MaSp fiber is 0.001 to 5.

10. The derivatized porous MaSp fiber according to claim 1, wherein the functional portion is further bonded to a dye or pigment.

11. The derivatized porous MaSp fibers have a decomposition temperature (T) of 280°C to 350°C, as determined by differential scanning calorimetry (DSC). ｄ ), and a glass transition temperature of 200°C to 250°C (T) as determined by DSC. ｇ A derivatized porous MaSp fiber according to claim 1, characterized by the following:

12. The derivatized porous MaSp-based fiber has a repeating region containing the amino acid sequence shown in Formula 10: (X １ ), Ｚ X ２ GPGGYGPX ３ X ４ X ５ GPX ６ GX ７ GGX ８ GPGGPGX ９ X １０ wherein, in each case independently, X １ is A or G, Z is an integer from 5 to 30, X ２ is S or G, X ３ is G or E, X ４ is G, S or N, X ５ is Q or Y, X ６ is G or S, X ７ is P or R, X ８ is Y or Q, X ９ is G or S, and X １０ is S or G. The derivatized porous MaSp-based fiber according to claim 1.

13. A composite material comprising a derivatized porous MaSp fiber according to claim 1, bonded to one or any combination thereof of a metal, a salt thereof, and metal oxide particles.

14. The composite material according to claim 13, wherein the metal, its salt, or the metal oxide particles are bonded to the derivatized porous MaSp fibers via a chelating agent.

15. The composite material according to claim 14, wherein the chelating agent comprises (i) a metal chelating group that can bond to a metal or a salt thereof, (ii) a metal oxide chelating group, or both (i) and (ii).

16. The composite material according to claim 15, wherein the metal chelating group comprises iminodiacetic acid (IDA), DOTA, NOTA, NODA, EDTA, HBED-CC (including any salts, derivatives, or combinations thereof), and the metal oxide chelating group is selected from salicylic acid, phosphonic acid, hydroxamic acid, malonic acid, pyrogallol, and 5-hydroxy-1,4-naphthoquinone (including any salts or combinations thereof).

17. The composite material according to claim 14, wherein the molar ratio of the chelating agent to the derivatized porous MaSp fiber is 0.01 to 1.

18. The composite material according to claim 13, wherein the metal oxide particles are selected from titania, zirconia, silica, or any combination thereof.

19. The composite material according to claim 13, wherein the metal oxide particles are characterized by having a particle size of 10 to 5,000 nm.

20. The composite material according to claim 13, wherein the w / w ratio of the derivatized porous MaSp fibers to the metal oxide particles in the composite material is 0.01 to 100.

21. A method for synthesizing a derivatized porous MaSp fiber according to claim 1, wherein the porous MaSp fiber is given formula I: 【Transformation 3】 Or formula II: 【Chemistry 4】 The process includes reacting with a reagent represented by the formula, where, R is a functionalized moiety or polymer as described in claim 1, and A is selected from substituted or unsubstituted aryl, heteroaryl, and alkyl groups, including any combination thereof, and each R １ A method for obtaining the derivatized porous MaSp fiber, wherein each component independently comprises one of hydrogen, alkyl, hydroxy, or alkoxy (including any combination thereof), the wavy bond represents a linker or bond, and the reaction step includes conditions sufficient to covalently bond the functional portion to the tyrosine of the porous MaSp fiber.

22. The method according to claim 21, wherein the covalent bond is mediated by a diazo bond or C-Si.

23. R １ Ha-O-C １～１０ The method according to claim 21 or 22, comprising an alkyl group.

24. The method according to claim 21, wherein A comprises a substituted or unsubstituted phenyl.

25. The linker is substituted or non-substituted C １～１０ The method according to claim 21, comprising an alkyl group.

26. The method according to claim 21, wherein R is the polymer, the method further comprises the step of reacting the functionalized portion with the polymer to covalently bond the polymer to the porous MaSp fiber, the functionalized portion comprises an amine or a carboxyl group, the polymer is reactive to the functionalized portion, and the linker comprises optionally substituted alkyl, amino acid, alkoxy, thioalkyl, aminoalkyl, glycol, -C(O)NH-, -C(O)O-, -C(O)-, -C(O)S-, -C(NH)NH-, -C(NH)O-, -C(NH)S-, urea, or any combination thereof.

27. The method according to claim 26, further comprising the step of reacting the polymer with a chelating agent having reactivity with the polymer to obtain the chelating agent covalently bonded to the polymer.

28. The method according to claim 27, wherein the polymer comprises PGA.

29. An article comprising the derivatized porous MaSp fiber described in claim 21 or the composite material described in claim 13.

30. The article according to claim 29, wherein the article is a conductive article, an antimicrobial article, or a cosmetic article (including any combination thereof).

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