Antibacterial nanoworms

Abstract

At least one nanoworm comprises a plurality of alkene units and a plurality of macro-CTA polymer units, the macro-CTA polymer units comprising R 1 from a reversible addition-fragmentation chain transfer agent. 1 In some embodiments, the R 1 The groups may be functional groups such as, for example, carboxylic acids, alkynes, pyridines, dopamines, thiolactones, biotin, azides, peptide sequences, sugar sequences, proteases, glycanases, polymers, other functional groups, and combinations thereof. In some embodiments, the macroCTA polymer units comprise quaternized amines. In some embodiments, the macroCTA polymer units comprise functionalized quaternized amines, such as alkyl groups, carboxylic acids, alkynes, pyridines, dopamines, thiolactones, biotin, azides, peptide sequences, sugar sequences, proteases, glycanases, polymers, other functional groups, and combinations thereof. In some embodiments, the coating comprises at least one nanoworm.

Description

[Technical Field]

[0001] Cross-reference of related applications

[0001] This application claims priority to U.S. Provisional Patent Application No. 62 / 899,983 filed on 13 September 2019 and U.S. Provisional Patent Application No. 63 / 049,813 filed on 9 July 2020, both of which are incorporated herein by reference in their entirety.

[0002] Technical field

[0002] The present disclosure provides antimicrobial nanoworms such as coatings of antimicrobial nanoworms on the surface of transporters, buildings, wearables, filters, or any other object. [Background technology]

[0003]

[0003] Pandemics caused by viruses (such as SARS, SARS-CoV-2, swine flu, and Ebola) have a significant impact on the global economy by reducing the number of passengers on airlines. Seasonal influenza and sterilization of aircraft cabins are also ongoing concerns for airline passengers. Similarly, the space transport and habitation industries are concerned with preventing the transmission of microorganisms. People traveling in space may be more susceptible to the transmission of microbial diseases and may be more prone to immunosuppression. Furthermore, in a weightless or radiation-shielded environment, microorganisms may replicate more easily and become more virulent.

[0004]

[0004] Preventing disease transmission in aircraft and spacecraft has traditionally focused on improving air filtration systems, such as HEPA air filter systems. Replacing and maintaining HEPA filters can be costly or impractical, such as replacing and maintaining HEPA filters in space. Furthermore, such systems may not be effective in reducing or preventing the transmission of microorganisms from surfaces. Bacteria and viruses can remain on surfaces for several days to up to a week.

[0005]

[0005] Therefore, an antimicrobial surface coating is needed that is effective in reducing the transmission of microorganisms. [Overview of the project]

[0006]

[0006] The present disclosure provides nanoworms such as antimicrobial nanoworm coatings on the surface of transporters, buildings, wearables, filters, or any other object.

[0007]

[0007] At least one nanoworm comprises a plurality of alkene units and a plurality of macro CTA polymer units. The macro CTA polymer units are derived from a reversible addition-cleavage chain transfer agent. 1 Contains a group. In one embodiment, the macro CTA polymer unit R 1 The groups are, for example, functional groups such as carboxylic acids, alkynes, pyridines, dopamine, thiolactones, biotin, azides, peptide sequences, sugar sequences, proteases, glycanases, polymers, other functional groups, and / or combinations thereof. In some embodiments, the macro-CTA polymer unit comprises a quaternized amine. In some embodiments, the macro-CTA polymer unit comprises a functionalized quaternized amine, for example, an alkyl group, a carboxylic acid, an alkyne, a pyridine, dopamine, thiolactone, biotin, azide, a peptide sequence, a sugar sequence, a protease, a glycanase, a polymer, other functional groups, and / or combinations thereof. In some embodiments, the coating comprises at least one nanoworm.

[0008]

[0008] At least one nanoworm comprises a plurality of alkene units, a first set of a plurality of macro-CTA polymer units, and a second set of a plurality of macro-CTA polymer units. The macro-CTA polymer units of the first set and the second set are derived from a reversible addition-cleavage chain transfer agent. 1 Includes a group. The first set of multiple macro CTA polymer units is different from the second set of multiple macro CTA polymer units. In one embodiment, the R of the macro CTA polymer units 1The groups are, for example, functional groups such as carboxylic acids, alkynes, pyridines, dopamine, thiolactones, biotin, azides, peptide sequences, sugar sequences, proteases, glycanases, polymers, other functional groups, and / or combinations thereof. In some embodiments, the macro-CTA polymer unit comprises a quaternized amine. In some embodiments, the macro-CTA polymer unit comprises a functionalized quaternized amine, for example, an alkyl group, a carboxylic acid, an alkyne, a pyridine, dopamine, thiolactone, biotin, azide, a peptide sequence, a sugar sequence, a protease, a glycanase, a polymer, other functional groups, and / or combinations thereof. In some embodiments, the coating comprises at least one nanoworm.

[0009]

[0009] At least one nanoworm comprises a plurality of alkene units, a plurality of macro-CTA polymer units, and a plurality of grafted polymers. The first set of macro-CTA polymer units are derived from a reversible addition-cleavage chain transfer agent. 1 The group is included. Multiple grafted polymers are grafted onto at least a portion of a first set of multiple macro-CTA polymer units. In one embodiment, the R of the macro-CTA polymer units 1 The groups include, for example, carboxylic acids, alkynes, pyridines, dopamine, thiolactones, biotin, azides, peptide sequences, sugar sequences, proteases, glycanase polymers, polymers, grafted polymers, other functional groups, and / or combinations thereof. In some embodiments, the macro-CTA polymer unit and / or grafted polymer unit comprises a quaternized amine. In some embodiments, the macro-CTA polymer unit and / or grafted polymer comprises a functionalized quaternized amine, for example, alkyl groups, carboxylic acids, alkynes, pyridines, dopamine, thiolactones, biotin, azides, peptide sequences, sugar sequences, proteases, glycanase polymers, other functional groups, and / or combinations thereof. In some embodiments, the macro-CTA polymer unit comprises a functionalized quaternized amine containing a grafted polymer. In some embodiments, the coating comprises at least one nanoworm.

[0010]

[0010] To enable a detailed understanding of the above-mentioned features of the Disclosure, a more specific description of the Disclosure, which has been briefly summarized above, can be obtained by referring to the embodiments, some of which are illustrated in the accompanying drawings. However, it should be noted that the accompanying drawings illustrate only typical embodiments of the Disclosure, as the Disclosure allows for other equally valid embodiments, and therefore should not be considered to limit the scope of the Disclosure. [Brief explanation of the drawing]

[0011] [Figure 1]

[0011] Figure 1 is a schematic diagram showing a nanoworm according to a certain embodiment. [Figure 2A-2B]

[0012] This is a schematic diagram showing a nanoworm having an R1 group containing a polymer, which has been further functionalized according to a certain embodiment. [Figure 3]

[0013] This is a schematic diagram showing a nanoworm having a quaternized amine group according to a certain embodiment. [Figure 4A-4B]

[0014] This is a schematic diagram showing a nanoworm having a quaternary amine group containing a polymer, which has been further functionalized according to a certain embodiment. [Figure 5]

[0015] This is a schematic cross-sectional view of a coating containing multiple nanoworms on top of a substrate, according to one embodiment. [Figure 6]

[0016] This is a schematic cross-sectional view of a coating comprising a plurality of nanoworms on a substrate having a first set of macro CTA polymer units having different LCSTs and a second set of macro CTA polymer units, according to one embodiment. [Figure 7]

[0017] This graph shows an example of determining the LCST of macro CTA in water at different weight fractions and pH levels according to a certain method. [Figure 8]

[0018] This graph shows an example of incorporating cationic and hydrophobic moieties into macro CTA in a certain manner. [Figure 9]

[0019] Examples of LCST profiles of macro CTA after quaternization with iodomethane and iodooctane at different pH levels, according to a certain embodiment, are shown. [Figure 10]

[0020] An example of the antimicrobial activity against E. coli on a glass surface coated with nanoworms, according to one embodiment, is shown. [Figure 11]

[0021] An example of antimicrobial activity against the H3N2 influenza virus on a surface coated with nanoworms, according to one embodiment, is shown. [Figure 12]

[0022] An example of antimicrobial activity against the AAV-HA virus on a surface coated with nanoworms, according to a certain embodiment, is shown. [Figure 13]

[0023] An example of antimicrobial activity against the AAV-HA virus on a surface coated with nanoworms, according to a certain embodiment, is shown. [Figure 14]

[0024] This is a schematic diagram showing a nanoworm having two different R1 groups according to a certain embodiment. [Figure 15A]

[0025] A schematic diagram is shown illustrating a synthetic alkyne-γ-thiolactone PDMAEMA nanoworm quaternized with 10% equivalent iodooctane. [Figure 15B]

[0026] A schematic diagram is shown illustrating synthetic alkyne-γ-thiolactone PDMAEMA nanoworms quaternized with 10% equivalent iodooctane and 90% equivalent iodomethane. [Figure 15C]

[0027] A schematic diagram is shown illustrating synthetic alkyne-γ-thiolactone PDMAEMA nanoworms quaternized with an equivalent amount of propargyl bromide. [Figure 16A]

[0028] A schematic diagram shows a synthesized conjugated guanidine azide on a quaternized alkyne-γ-thiolactone PDMAEMA nanoworm, which contains approximately 10% tertiary amine groups quaternized by octyl groups. [Figure 16B]

[0029] A schematic diagram shows a synthesized conjugated guanidine azide in a quaternized alkyne-γ-thiolactone PDMAEMA nanoworm, which contains approximately 10% tertiary amine groups quaternized with octyl groups and approximately 90% tertiary amines quaternized with methyl groups. [Figure 17A]

[0030] A schematic diagram is shown illustrating quaternized P(NIPAM55-co-DMAEMA48) grafted onto an alkyne-γ-thiolactone PDMAEMA nanoworm. [Figure 17B]

[0031] A schematic diagram is shown illustrating P(NIPAM55-co-DMAEMA48), which is produced as a result of grafting alkyne-γ-thiolactone PDMAEMA nanoworms. [Figure 17C]

[0032] A schematic diagram is shown illustrating the conjugation resulting from guanidine azide to quaternized grafted alkyne PDMAEMA nanoworms. [Figure 17D]

[0033] A schematic diagram is shown illustrating the resulting conjugation of guanidine azide and polygalactose azide onto quaternized grafted alkyne PDMAEMA nanoworms. [Figure 18-21]

[0034] This shows a decrease in viral activity when applied to various surfaces. [Modes for carrying out the invention]

[0012]

[0035] To facilitate understanding, the same reference numerals were used where possible to indicate identical elements common to multiple figures. The figures are not drawn to scale and may be simplified for clarity. Elements and features of one embodiment are considered to be advantageously incorporated into other embodiments without further explanation.

[0013]

[0036] The use of the term "comprising" in the summary of the invention or the modes for carrying out the invention shall mean "comprising," "consisting essentially," and / or "consisting of."

[0014]

[0037] This disclosure provides nanoworms such as antimicrobial nanoworm coatings on the surface of transporters, buildings, wearables, filters, or any object. The nanoworms have antimicrobial properties that are effective in reducing or killing microorganisms and / or reducing microbial transmission. Microorganisms may be viruses, bacteria, fungi, and / or other microorganisms.

[0015]

[0038] In some embodiments, a surface coated with nanoworms is or can become hydrophilic. For example, a surface coated with nanoworms is or can become hydrophilic, allowing droplets such as mucosal droplets, blood, urine, sweat, other bodily fluids, and other non-bodily fluids to be wetted across the nanoworm-coated surface. In some embodiments, microorganisms on the surface of a droplet or suspended within a droplet can be captured and / or killed by the nanoworm-coated surface.

[0016]

[0039] In some embodiments, a surface coated with nanoworms is affected by the environmental conditions of the droplet and the surrounding external conditions. For example, a surface coated with nanoworms can be affected by one or more environmental factors such as temperature, pH, salinity, and / or light, which can help capture and kill microorganisms. For example, when a droplet evaporates, the surface coated with nanoworms can change from hydrophilic to water-insoluble. This change from hydrophilic to water-insoluble can enhance the capture and killing of microorganisms within the droplet. For example, one or more nanoworms on a surface coated with nanoworms may have multiple adhesion or contact points with microorganisms, where the change from hydrophilic to water-insoluble can exert mechanical strain on the microorganisms, causing them to dissociate or decompose.

[0017]

[0040] In some embodiments, the chemical composition and functionality of multiple nanoworms may be selected to enhance the capture and killing of microorganisms. For example, a surface coated with nanoworms may be chemically modified with carboxylic acid groups, alkynes, pyridines, dopamines, thiolactones, biotins, azides, peptide sequences (including one or more amino acids and / or combinations thereof), nucleic acid sequences (including one or more nucleic acids and / or combinations thereof), sugar sequences (including one or more monosaccharides, polysaccharides and / or combinations thereof), proteases, glycanases, grafted polymers, quaternized amine groups, derivatives thereof, and / or combinations thereof to capture / kill a wide range of microorganisms or specific microorganisms, for example, depending on a particular outbreak of a virus.

[0018]

[0041] Nanoworm coatings can be non-toxic. For example, a surface coated with nanoworms is not toxic to humans, animals, and / or plants, and is antimicrobial. For instance, the antimicrobial compounds on a nanoworm-coated surface are covalently bonded to the nanoworms. Because the nanoworms adhere strongly to the surface, the antimicrobial compounds on the nanoworm-coated surface are prevented from being ingested or absorbed by the human body through the skin.

[0019]

[0042] In one aspect, the nanoworm coating can be washable because it can be washed and reused. For example, a surface coated with nanoworms can be washed with water (e.g., rinsing), a cleaning agent (detergent, soap, surfactant, etc.), a disinfectant, and / or a bactericide. By washing the surface coated with nanoworms to remove the captured or killed microorganisms from the antibacterial compounds of the nanoworms, the surface coated with nanoworms can be made new. The newly made nanoworms can capture and kill additional microorganisms on the surface coated with nanoworms. For example, the antibacterial compounds can be selected to capture and kill without covalently bonding to the microorganisms. Thus, by washing the nanoworms, the captured or killed microorganisms are released from the antibacterial compounds, enabling the antibacterial compounds to be made new for the capture and killing of additional microorganisms.

[0020]

[0043] In one aspect, the nanoworm comprises a copolymer of macro chain transfer agent (macro CTA) polymer units and alkene units. The macro CTA polymer is a polymer formed by reversible addition fragmentation chain transfer (RAFT) using a RAFT agent in the polymerization of one or more ethylenically unsaturated monomers. In one aspect, the RAFT agent is incorporated into the macro CTA polymer, which can be further polymerized by the addition of reactants.

[0021]

[0044] In a further aspect, the RAFT agent has the general formula (I): TIFF0007872223000001.tif30170[where R 1 is an x-valent group and x is an integer ≧1]. R 1 groups can be monovalent, divalent, trivalent, or higher valent. In one aspect, x is an integer in the range of 1 to 20, such as 1 to 10 or 1 to 5. Thus, R 1 is optionally a polymer that may be substituted, and the remainder of the RAFT agent is represented as a multi-component group pendant from the polymer chain. R 1The group may be an organic group or a substituted organic group that functions as a free radical leaving group under the polymerization conditions used. The Z group can be independently selected from organic groups and / or substituted organic groups that give the C=S moiety of the RAFT agent a sufficiently high reactivity to free radical addition.

[0022]

[0045] R in equation (I) 1 Examples of which may be substituted include alkyl, alkenyl, alkynyl, aryl, acyl, carbocykyl, heterocyclyl, heteroaryl, alkylthio, alkenylthio, alkynylthio, arylthio, acylthio, carbocykrillthio, heterocyclylthio, heteroarylthio, alkylalkenyl, alkylalkynyl, alkylaryl, alkylacyl, alkylcarbocykyl, alkylheterocyclyl, alkylheteroaryl, alkyloxyalkyl, alkenyloxyalkyl, alkynyloxyalkyl, aryloxyalkyl, alkylacyloxy, alkylcarbocykyloxy, alkylheterocyclyloxy, alkylheteroaryloxy, alkylthioalkyl, alkenylthioalkyl, alkynylthioalkyl, arylthioalkyl This includes alkylacylthio, alkylcarbocyclylthio, alkylheterocyclylthio, alkylheteroarylthio, alkylalkenylalkyl, alkylalkynylalkyl, alkylarylalkyl, alkylacylalkyl, arylalkylaryl, arylalkenylaryl, arylalkynylaryl, arylacylaryl, arylacyl, arylcarbocyclyl, arylheterocyclyl, arylheteroaryl, alkenyloxyaryl, alkynyloxyaryl, aryloxyaryl, alkylthioaryl, alkenylthioaryl, alkynylthioaryl, arylthioaryl, arylacylthio, arylcarbocyclylthio, arylheterocyclylthio, arylheteroarylthio, and polymer chains.

[0023]

[0046] R in equation (I) 1Examples include optionally substituted alkyl groups, saturated, unsaturated, or aromatic carbocyclic or heterocyclic groups, alkylthio groups, dialkylamino groups, organometallic species, and polymer chains.

[0024]

[0047] R in equation (I) 1 In certain examples, C1-C may be substituted. 18 Alkyl, C2-C 18 Alkenyl, C2-C 18 Alkinyl, C6-C 18 Ariel, C1-C 18 Ashiru, C3-C 18 Carbocyclyl, C2-C 18 Heterocyclyl, C3-C 18 Heteroaryl, C1-C 18 Alkylthio, C2-C 18 Alkenylthio, C2-C 18 Alkinylthio, C6-C 18 Arylthio, C1-C 18 Asyltio, C3-C 18 Carbocyclilthio, C2-C 18 Heterocyclilthio, C3-C 18 Heteroarylthio, C3-C 18 Alkyl alkenyl, C3-C 18 Alkylalkynyl, C7-C 24 Alkylaryl, C2-C 18 Alkylacyl, C4-C 18 Alkylcarbocyryl, C3-C 18 Alkyl heterocyclyl, C4-C 18 Alkyl heteroaryl, C2-C 18 Alkyloxyalkyl, C3-C 18 Alkenyloxyalkyl, C2-C 18 Alkynyloxyalkyl, C7-C 24 Aryloxyalkyl, C2-C 18 Alkyl acyloxy, C2-C 18 Alkylthioalkyl, C3-C 18 Alkenylthioalkyl, C3-C 18 Alkinylthioalkyl, C7-C 24Arylthioalkyl, C2-C 18 Alkyl acylthio, C4-C 18 Alkylcarbocycrylthio, C3-C 18 Alkyl heterocyclilthio, C4-C 18 Alkyl heteroarylthio, C4-C 18 Alkylalkenylalkyl, C4-C 18 Alkylalkynylalkyl, C8-C 24 Alkylarylalkyl, C3-C 18 Alkyl acylalkyl, C 13 -C 24 Arylalkylaryl, C 14 -C 24 Arylalkenylaryl, C 14 -C 24 Arylalkynylaryl, C 13 -C 24 Aryl acyl aryl, C7-C 18 Arylacyl, C9-C 18 Arylcarbocykrill, C8-C 18 Aryl heterocyclyl, C9-C 18 Aryl heteroaryl, C8-C 18 Alkenyloxyaryl, C8-C 18 Alkynyloxyaryl, C 12 -C 24 Aryloxyaryl, C7-C 18 Alkylthioaryl, C8-C 18 Alkenylthioaryl, C8-C 18 Alkinylthioaryl, C 12 -C 24 Arylthioaryl, C7-C 18 Arylacylthio, C9-C 18 Arylcarbocycrine, C8-C 18 Aryl heterocyclilthio, C9-C 18 This includes aryl heteroarylthio and polymer chains having a number-average molecular weight in the range of about 500 to about 80,000, for example, in the range of about 500 to about 30,000.

[0025]

[0048] Examples of Z in formula (I) include F, Cl, Br, I, alkyl, aryl, acyl, amino, carbocykrill, heterocyclyl, heteroaryl, alkyloxy, aryloxy, acyloxy, acylamino, carbocykrilloxy, heterocyclyloxy, heteroaryloxy, alkylthio, arylthio, acylthio, carbocykrillthio, heterocyclylthio, heteroarylthio, alkylaryl, alkylacyl, alkylcarbocykrill, alkylheterocyclyl, alkylheteroaryl, alkyloxyalkyl, aryloxyalkyl, alkylacyloxy, alkylcarbocykrilloxy, alkylheterocyclyloxy, alkylheteroaryloxy, alkylthioalkyl, arylthioalkyl, alkylacylthio, alkylcarbocykrillthio, alkylheterocyclylthio, This includes alkylheteroarylthio, alkylarylalkyl, alkylacylalkyl, arylalkylaryl, arylacylaryl, arylacyl, arylcarbocykrill, arylheterocyclyl, arylheteroaryl, aryloxyaryl, arylacyloxy, arylcarbocykrilloxy, arylheterocyclyloxy, arylheteroaryloxy, alkylthioaryl, arylthioaryl, arylacylthio, arylcarbocykrillthio, arylheterocyclylthio, arylheteroarylthio, dialkyloxy-, diheterocyclyloxy- or diaryloxy-phosphinyl, dialkyl-, diheterocyclyl- or diaryl-phosphinyl, cyano (i.e., -CN), and -SR (where R is as defined with respect to formula (I)).

[0026]

[0049] Specific examples of Z in formula (I) include F, Cl, and C1-C which may be substituted. 18 Alkyl, C6-C 18 Ariel, C1-C 18 Acyl, amino, C3-C 18 Carbocyclyl, C2-C 18 Heterocyclyl, C3-C 18 Heteroaryl, C1-C 18 Alkyloxy, C6-C 18Aryloxy, C1-C 18 Acryloxy, C3-C 18 Carbocyclryloxy, C2-C 18 Heterocyclyloxy, C3-C 18 Heteroaryloxy, C1-C 18 Alkylthio, C6-C 18 Arylthio, C1-C 18 Acrylthio, C3-C 18 Carbocyclrylthio, C2-C 18 Heterocyclylthio, C3-C 18 Heteroarylthio, C7-C 24 Alkylaryl, C2-C 18 Alkylacyl, C4-C 18 Alkylcarbocyclryl, C3-C 18 Alkylheterocyclyl, C4-C 18 Alkylheteroaryl, C2-C 18 Alkyloxyalkyl, C7-C 24 Aryloxyalkyl, C2-C 18 Alkylacryloxy, C4-C 18 Alkylcarbocyclryloxy, C3-C 18 Alkylheterocyclyloxy, C4-C 18 Alkylheteroaryloxy, C2-C 18 Alkylthioalkyl, C7-C 24 Arylthioalkyl, C2-C 18 Alkylacrythio, C4-C 18 Alkylcarbocyclrylthio, C3-C 18 Alkylheterocyclylthio, C4-C 18 Alkylheteroarylthio, C8-C 24 Alkylarylalkyl, C3-C 18 Alkylacylalkyl, C 13 -C 24 Arylalkylaryl, C 13 -C 24 Arylacylaryl, C7-C 18 Arylacyl, C9-C 18 Arylcarbocyclryl, C8-C 18 Arylheterocyclyl, C9-C18 Arylheteroaryl, C 12 -C 24 Aryloxyaryl, C7-C 18 Aryl acyloxy, C9-C 18 Arylcarbocyroxy, C8-C 18 Aryl heterocyclyloxy, C9-C 18 Aryl heteroaryloxy, C7-C 18 Alkylthioaryl, C 12 -C 24 Arylthioaryl, C7-C 18 Arylacylthio, C9-C 18 Arylcarbocycrine, C8-C 18 Aryl heterocyclilthio, C9-C 18 Arylheteroarylthio, dialkyloxy-, diheterocyclyloxy- or diaryloxy-phosphinyl (i.e., -P(=O)OR k 2) Dialkyl-, diheterocyclyl-, or diaryl-phosfinyl, (i.e., -P(=O)R k 2)(R k C1-C may be substituted. 18 Alkyl, possibly substituted C6-C 18 Aryl, possibly substituted C2-C 18 Heterocyclyl, and C7-C which may be substituted. 24 These include alkylaryls (selected from alkylaryls), cyanos (i.e., -CN), and -SRs (where R is as defined with respect to formula (I)).

[0027]

[0050] R 1 In the example of Z, the multicomponent group includes subgroups in any order. For example, the multicomponent group of alkylaryl includes arylalkyl. 1 Alternatively, Z may be branched and / or substituted. 1 Alternatively, Z may contain an alkyl moiety that is substituted, and any substituents may be -O-, -S-, -NR a-, -C(O)- (i.e., carbonyl), -C(O)O- (i.e., ester), and -C(O)NR a - This includes cases where it is replaced by a group selected from (i.e., an amide), where R a The elements are selected from halogens, alkyls, alkenyls, alkynyls, aryls, carbocyrills, heteroaryls, heterocyclyls, arylalkyls, and acyls.

[0028]

[0051] In this specification, references to the x-valent, polyvalent, or divalent "forms of..." are intended to mean that a particular group is an x-valent, polyvalent, or divalent radical, respectively. For example, when x is 2, a particular group is intended to be a divalent radical. In this case, a divalent alkyl group is effectively an alkylene group (e.g., -CH2-). Similarly, the divalent form of an alkylaryl group can be represented, for example, -(C6H4)-CH2-, a divalent alkylarylalkyl group can be represented, for example, -CH2-(C6H4)-CH2-, a divalent alkyloxy group can be represented, for example, -CH2-O-, and a divalent alkyloxyalkyl group can be represented, for example, -CH2-O-CH2-. When the term "optionally substituted" is used in combination with such x-valent, polyvalent, or divalent, the group can be substituted or condensed as described herein. If an x-valent group, polyvalent group, or divalent group contains two or more subgroups, for example [group A][group B][group C] (e.g., alkylarylalkyl), then one or more of these subgroups may be substituted.

[0029]

[0052] In one embodiment, some or all of the RAFT agent is incorporated into the macro-CTA polymer. In another embodiment, R 1And -S-(S=O)-Z are incorporated into the macro-CTA polymer. Examples of RAFT polymerized macro-CTAs include, but are not limited to, poly(N-isopropylacrylamide) (PNIPAM), poly(N,N-(dimethylamino)ethyl methacrylate) (F), poly(N-acetoxyethylacrylamide) (PNAEAA), poly(acryloylglycine ethyl ester) (PNAGEE), poly((ethylene glycol)methyl ether methacrylate) (PEGMEMA), poly((propylene glycol) methacrylate) (PPGMA), and poly(N,N-dimethyl This includes acrylamide (PDMA), poly(N-decylacrylamide) (PDcA), poly(N,N-diethylacrylamide) (PDEA), poly(N-acryloylglycine) (PNAG), poly(N-acryloylglycine methyl ester) (PNAGME), poly(N-acryloylglycine ethyl ester) (PNAGEE), and poly(N-acryloylglycine propyl ester) (PNAGPE), other polyacrylamides, other polyacrylates, and copolymers thereof.

[0030]

[0053] For example, macro CTAs containing poly(NIPAM) are given by general formula (II): TIFF0007872223000002.tif39170Z-(C=S)-S-(NIPAM) x -R 1 (II) [In the formula, Z and R 1 (Functionalization or Functionalization R 1 The RAFT agent has a component (including the base). In one embodiment, x is any positive integer. In another embodiment, x is an integer from 10 to 100.

[0031]

[0054] For example, macro CTAs containing poly(NIPAM-co-DMAEMA) are given by general formula (III): TIFF0007872223000003.tif68170Z-(C=S)-S-[(NIPAM) x -(DMAEMA) y ]-R 1 (III) [In the formula, Z and R 1 (Non-functionalization or functionalization R 1 The group (including) is a component of the RAFT agent. In some embodiments, x and y are any independently selected positive integers. In some embodiments, x is an integer from 10 to 100 and y is an integer from 10 to 100. The sequence of monomers of the macro-CTA copolymer, such as the NIPAM and DMAEMA monomers of the macro-CTA copolymer of formula (III), can be any sequence, such as a random sequence, an alternating sequence, a statistical sequence, a periodic sequence, or a block sequence.

[0032]

[0055] In one embodiment, macro-CTA polymers are used to form nanoworms by further polymerizing macro-CTA polymer units and alkene monomers. For example, nanoworms can be produced by first generating macro-CTA polymer units using a RAFT agent in which the RAFT agent is incorporated into each macro-CTA polymer unit. The macro-CTA polymer units and alkene monomers are polymerized together to form nanoworms. The RAFT agent incorporated into the macro-CTA polymer polymerizes the alkene units and macro-CTA units to form a nanoworm with general formula (IV): (Alkene units) m (Macro CTA unit) n (IV) [In the formula, each of the macro CTAs is either non-functionalized or functionalized as a component of the RAFT agent.] 1 The nanoworm is formed containing the group. The nanoworm contains macro CTA units, alkene units, and components of the RAFT agent. The alkene units and macro CTA units can be formed within the nanoworm in any sequence, such as random, alternating, statistical, periodic, or block sequences.

[0033]

[0056] Polyalkenes are formed by polymerization of any suitable alkene monomer and / or combinations thereof. Examples of suitable alkene monomers include ethylene, propylene, 1-butene, 1-pentene, 4-methyl-1-pentene, 1-hexene, 1-heptene, 1-octene, 1-decene, norbornene, styrene, acrylates, methacrylates, other vinyl compounds, their substituted compounds, and their derivatives. In some embodiments, the polyalkene unit of a nanoworm contains m monomer units, such as 20 to 400 alkene monomer units, e.g., 25 to 35 alkene monomer units. In some embodiments, the polyalkene contains polystyrene. In some embodiments, m and n are any independently selected positive integers. In some embodiments, the macro CTA unit of a nanoworm contains n macro CTA units, such as 1 to 200, e.g., 2 to 100 macro CTA units.

[0034]

[0057] For example, a nanoworm containing polystyrene and a macro CTA of poly(NIPAM-co-DMEA) has the general formula (V): TIFF0007872223000004.tif64170 (Styrene) m [(NIPAM) x -(DMAEMA) y -R 1 ] n (V) [In the formula, R 1 This refers to non-functionalized or functionalized R as a component of RAFT agents. 1 It has a base. In one embodiment, x, y, m, and n are any independently chosen positive integers. In one embodiment, x is an integer from 10 to 100, y is an integer from 10 to 100, m is an integer from 20 to 400, and n is an integer from 1 to 200.

[0035]

[0058] Macro CTA polymer units may be selected to provide properties to nanoworms. Macro CTA polymer units may be configured to respond to temperature, pH, salinity, light, and / or combinations thereof. For example, macro CTA polymer units can change their miscibility with droplets based on temperature alone, or based on temperature, pH, salinity, light, and / or other external environmental conditions.

[0036]

[0059] In some embodiments, macro-CTA polymer units may contain temperature-responsive monomers and / or functional groups in any appropriate amount. Temperature-responsive macro-CTA polymer units may have an LCST, an upper critical eutectic temperature (UCST), or both an LCST and an UCST. Examples of temperature-responsive monomers or functional groups include those having amine functional groups, carbonyl functional groups, and / or combinations thereof. In some embodiments, macro-CTA polymer units have an LCST in water from about -20°C to about +100°C.

[0037]

[0060] In some embodiments, macro CTA units may contain pH-responsive monomers and / or functional groups in any appropriate amount. Examples of pH-responsive monomers include vinyl monomers, e.g., acrylic acid, methacrylic acid, and other alkyl-substituted acrylic acids, maleic anhydride, maleic acid, 2-acrylamido-2-methyl-l-propanesulfonic acid, N-vinyl formaldehyde, N-vinyl acetamide, aminoethyl methacrylate, phosphoryl ethyl acrylate, or methacrylate. Other examples of pH-responsive monomers include amino acid-derived polypeptides (e.g., polylysine and polyglutamic acid), polysaccharides (e.g., alginic acid, hyaluronic acid, carrageenan, chitosan, carboxymethyl, and cellulose), or nucleic acids (e.g., deoxyribonucleic acid (DNA), ribonucleic acid (RNA), messenger RNA (mRNA), and their fragments). Examples of pH-sensitive functional groups include, but are not limited to, -OPO(OH)2, -COOH, or -NH2.

[0038]

[0061] In some embodiments, macro-CTA polymer units may contain salt-responsive monomers and / or functional groups in any appropriate amount. Examples of salt-responsive monomers and / or functional groups include ureidoamides, amines, carboxylic acid side groups, and other functional groups. Examples of salt-responsive macro-CTA polymer units include LCST polymers and / or UCST polymers.

[0039]

[0062] In some embodiments, macro CTA polymer units may contain photoresponsive monomers and / or functional groups in any appropriate amount. Examples of photoresponsive monomers and / or functional groups include those having chromophore functional groups. A chromophore functional group is any functional group sensitive to electromagnetic radiation (i.e., visible or invisible light). As used herein, “visible light” is defined as electromagnetic radiation having wavelengths from 380 nm to 750 nm. As used herein, “invisible light” is defined as electromagnetic radiation having wavelengths shorter than 380 nm (e.g., gamma rays, X-rays, ultraviolet rays) or longer than 750 nm (e.g., infrared rays, microwaves, radio waves). Examples of chromophore functional groups include groups that are trans-to-cis isomerized or capable of causing isomerization, and / or relatively nonpolar, hydrophobic groups that are capable of causing or capable of causing transitions from a non-ionized state to a hydrophilic ionic state, and / or groups polymerized with other monomers or comonomers in response to electromagnetic radiation. Examples of chromophore functional groups include azide-containing fluorescent dyes that can be functionalized to macro CTA polymer units via the CUAAC reaction, such as 3-azide-7-hydroxycoumarin, azide-BDP-FL, 5-FAM-azide, 6-FAM-azide, picolyl-azide-5 / 6-FAM, AF488-azide, AF488-picolyl-azide, 110-PEG3-azide, 5-SIMA-azide, 5-TAMRA-azide, 5 / 6-TAMRA-PEG3-azide, picolyl-azide-5 / 6-TAMRA, Cy3-azide, and sulfo. This includes -Cy3-azid, picolyl-azid-sulfo-Cy3, AF546-azid, AF546-picolyl-azid, AF555-azid, AF555-picolyl-azid, 5 / 6-Texas Red-PEG3-azid, AF594-azid, AF594-picolyl-azid, Cy5-azid, sulfo-Cy5-azid, picolyl-azid-sulfo-Cy5, AF647-azid, AF647-picolyl-azid, Cy5.5-Azid, picolyl-azid-Cy5.5, Cy7-azid, and picolyl-azid-Cy7.

[0040]

[0063] R of RAFT agent 1 The base, macro CTA polymer R1 Base, or R of nanoworm 1 The group can be pre-functionalized or post-functionalized. For example, the R of nanoworms. 1 The base is the R of RAFT agents. 1 Functionalizing the group, or the R of macro CTA polymers 1 By functionalizing the group, post-functionalization can be performed after the formation of nanoworms, and pre-functionalization can be performed before the formation of nanoworms. For example, the R of macro CTA polymers 1 The base is the R of RAFT agents. 1 By functionalizing the groups, post-functionalization can be performed after the formation of the macro-CTA polymer, and pre-functionalization can be performed before the formation of the macro-CTA polymer.

[0041]

[0064] RAFT agent, macro CTA polymer, and / or nanoworm R 1 The group may be functionalized to include carboxylic acid groups, alkynes, pyridines, dopamine, thiolactones, biotin, azides, peptide sequences, nucleic acid sequences, sugar sequences, proteases, glycans, polymers, chromophore functional groups, other functional groups, derivatives thereof, and / or combinations thereof. RAFT agents, macro-CTA polymers, and / or nanoworms 1 Specific examples of the basis include, but are not limited to, the following: TIFF0007872223000005.tif211170TIFF0007872223000006.tif75170 The functional group may be unsubstituted or substituted, non-halogenated or halogenated, or derivatives thereof.

[0042]

[0065] In some embodiments, R 1The group is functionalized to contain a peptide sequence. The peptide sequence contains one or more amino acids and / or one or more amino acid components. Examples of amino acid peptide sequences include, but are not limited to, GRGD (Gly-Arg-Gly-Asp), RGD (Arg-Gly-Asp), and other peptide sequences. Examples of amino acid peptide sequences include glycoproteins such as vivonectin, fibronectin, and other glycoproteins. Examples of amino acid components include guanidine, butylammonium, imidazolium, and other groups. In one example, a peptide sequence using a peptide azide contains an alkyne functional group R 1 It can be bound to RAFT agents, macro-CTA polymers, or nanoworms through a Cu(I)-catalyzed alkyne-azide (CuAAC) click reaction with the group. In another example, a peptide sequence using a peptide alkyne contains an azide functional group. 1 It can be bound to RAFT agents, macro-CTA polymers, or nanoworms through a CuAAC click reaction with a base.

[0043]

[0066] In some embodiments, R 1 The group is functionalized, for example, through a CuAAC click reaction, to include a sugar sequence. The sugar sequence includes one or more monosaccharides, polysaccharides, and / or combinations thereof. Examples of sugars include fucose, glucose, mannose, galactose, GalNac, GlcNAc, sialic acid, other glycans, other amino sugars, other acid sugars, their derivatives, their isomers, their polysaccharides, and / or combinations thereof. In some embodiments, unless expressly stated in the claims, the functionalized sugar sequence R attached to the glycoprotein of an enveloped virus is not bound by any particular theory. 1 The substance is thought to interfere with the glycosylation of enveloped viruses, prevent them from attaching to host cells, and / or prevent them from entering host cells.

[0044]

[0067] In some embodiments, R 1The base is functionalized, for example, through a CuAAC click reaction, to include a protease (referring to any compound that breaks down peptides or amino acids). Examples of proteases include common peptide denaturants and asparagine, serine, threonine, or specific peptide denaturants that target their binding. In some embodiments, unless expressly stated in the claims, the invention is not bound to any particular theory, but includes functionalized proteases R bound to the glycoprotein of enveloped viruses. 1 The compound is thought to interfere with the glycosylation of enveloped viruses, bind to the capsid of non-enveloped viruses, prevent the attachment of viruses (enveloped or non-enveloped) to host cells, and / or prevent the entry of viruses (enveloped or non-enveloped) into host cells.

[0045]

[0068] In some embodiments, R 1 The base is functionalized, for example, through a CuAAC click reaction, to include a glycanase (referring to any compound that breaks down glycans). Examples of glycans include common glycan denaturants and specific glycan denaturants that target the binding of fucose, glucose, mannose, galactose, GalNac, GlcNAc, sialic acid, or their binding. In some embodiments, unless expressly stated in the claims, the functionalized glycanase R bound to the glycoprotein of an enveloped virus is not bound to any particular theory. 1 The substance is thought to interfere with the glycosylation of enveloped viruses, prevent them from attaching to host cells, and / or prevent them from entering host cells.

[0046]

[0069] Figure 1 is a schematic diagram showing a nanoworm 100 in a certain embodiment. The skeleton or core 100 of the nanoworm 100 contains alkene units and macro CTA polymer units. The nanoworm 100 contains R derived from the macro CTA polymer units. 1 Includes base 130. R 1 Group 130 is a component of the RAFT agent and can be pre-functionalized or post-functionalized. 1Base 130 is any appropriate R 1 Includes the group R 1 The base may be selected to modify the capture and killing efficiency of the nanoworm 100, and / or to modify the responsiveness of the nanoworm 100 (e.g., temperature, pH, salinity, light, and / or combinations thereof).

[0047]

[0070] In one embodiment, nanoworms are formed by further polymerizing two or more sets of macro-CTA polymers and alkene monomers using two or more sets of macro-CTA polymer units. For example, the nanoworm is initially R 1 The base generates macro CTA-A units, R 1 It can be generated by generating macro CTA-B units from the base. R of macro CTA-A units 1 R of the base and macro CTA-B units 1 The groups may be the same or different. Macro CTA-A units, macro CTA-B units, and alkene monomers are polymerized together to form nanoworms. The RAFT agent incorporated within the macro CTA polymerizes the alkene units and macro CTA polymer units to form general formula (VI): (Alkene units) m (Macro CTA-A unit) n (Macro CTA-B unit) (VI) [In the formula, each of macro CTA-A is either non-functionalized or functionalized as a component of the RAFT agent.] 1 It contains a group, and each of macro CTA-B is either unfunctionalized or functionalized as a component of the RAFT agent. 1 The nanoworms include macro CTA-A units, macro CTA-B units, and components of the RAFT agent. In some embodiments, m, n, and o are any independently selected positive integers. In some embodiments, m is an integer from 20 to 400, n is an integer from 1 to 200, and o is an integer from 1 to 200. The alkene units, macro CTA-A units, and macro CTA-B units can be formed within the nanoworms in any sequence, such as random, alternating, statistical, periodic, or block sequences.

[0048]

[0071] Figure 14 shows two different R in one embodiment. 1 This is a schematic diagram showing a nanoworm 1400 having a group. The core 1410 of nanoworm 100 contains alkene units, macro CTA-A units, and macro CTA-B units. Nanoworm 1400 contains R derived from macro CTA-A units. 1 Contains base 1430-A and R derived from macro CTA-B unit 1 Includes base 1430-B. R 1 Groups 1430A and 1430B are components of the RAFT agent, and can be de-functionalized or functionalized (i.e., pre-functionalized or post-functionalized). 1 Base 1430A-B is R 1 Alkynes and other components of base 1430A and R 1 Any suitable R such as the B-thiolactone group of group 1430B 1 Includes a base (as shown in Figure 14). The same or different R of macro CTA-A and macro CTA-B. 1 The base may be selected to modify the capture and killing efficiency of the nanoworm 100, and / or to modify the responsiveness of the nanoworm 100 (e.g., temperature, pH, salinity, light, and / or combinations thereof).

[0049]

[0072] Figure 2A-2B shows R in a certain embodiment. 1 Figure 1 shows a schematic diagram of nanoworm 100, such as nanoworm 100 in Figure 1, in which group 130 is further functionalized to include polymer 150 (as shown in Figure 2B). Figures 2A-B use the same numbers as in Figure 1 for ease of explanation. As shown in Figure 2A, nanoworm 100 contains an alkyne group R 1 It contains group 130. As shown in Figure 2B, the polymer azide reacts with the alkyne group in Figure 2A via a Cu(I) catalyzed alkyne-azide (CuAAC) click reaction to functionalize the macro CTA polymer unit containing polymer 150 at its ends. 1Group 130 can be formed. For example, the polymer azide includes a peptide azide, a nucleic acid azide, a sugar azide, a protease azide, a glyconase azide, a polymer azide, or other polymer azides, and each forms a polymer 150 or other polymer containing a peptide sequence, a nucleic acid sequence, a sugar sequence, a protease, a glyconase, or a grafted polymer. In other embodiments, any polymer can be quaternized by an arbitrary reaction scheme, for example, by the reaction of macro-CTA polymer units with an azide group reacting with a polymer alkyne, thereby forming the R of macro-CTA polymer units. 1 It contains a group, or is bonded to one.

[0050]

[0073] Figure 3 is a schematic diagram showing a quaternized nanoworm 100 with amine groups 140, such as the nanoworm 100 in Figure 1, according to one embodiment. Figure 3 uses the same figures as Figure 1 for ease of explanation. One or more of the macro CTA polymer units have one or more tertiary amine groups. The tertiary amine groups are any appropriate R 2 It can be quaternized by the group. The R of the quaternized amine group 2 The groups may include or be functionalized to include alkyl, carboxylic acid, alkyne, pyridine, dopamine, thiolactone, biotin, azide, peptide sequence, nucleic acid sequence, sugar sequence, protease, glycanase, grafted polymer, chromophore functional group, other functional groups, derivatives thereof, and / or combinations thereof. For example, a macro CTA polymer unit may include or be functionalized to include two or more R groups to modify the capture and killing efficiency of nanoworms and / or to target common or specific microorganisms, including alkyl, carboxylic acid, alkyne, pyridine, dopamine, thiolactone, biotin, azide, peptide sequence, nucleic acid sequence, sugar sequence, protease, glycanase, grafted polymer, chromophore functional group, and other functional groups. 2 It may contain a group. The tertiary amine group is R 2 Reaction with halides or other R 2 It can be quaternized by a compound. 2 Specific examples of groups include alkyl moieties, alkyne moieties, and / or combinations thereof.

[0051]

[0074] In some embodiments, R 2 is an alkyl group of any suitable carbon length. Alkyl halides can quaternize tertiary amines to form the alkyl moiety. Nanoworms having macro CTA polymer units with multiple tertiary amine groups have alkyl groups in specific ratios of different groups. 2 It can be quaternized. The ratio of quaternized short-chain alkyl groups of 1 to 4 carbon atoms to long-chain alkyl groups of 5 or more carbon atoms can be selected to adjust the properties of Nanoworm 100. For example, the individual alkylamino groups of the monomers of the macro-CTA polymer units of Nanoworm 100 can be functionalized with quaternized methyl groups or quaternized octyl groups. For example, a Nanoworm containing macro-CTA polymer units of poly(NIPAM-co-DMAEMA) can be quaternized with z% long-chain alkyl groups (1-z%) and general formula (VII): TIFF0007872223000007.tif82170[where, R 1 (Non-functionalization or functionalization R 1 The group (including the group) may have a short-chain alkyl group which is a component of the RAFT agent. In one embodiment, a, b, x, y, m, and n are any independently selected positive integers, and z is independently any number between 0% and 100%. In one embodiment, x is an integer from 10 to 100, y is an integer from 10 to 100, z is a number between 0% and 100%, m is an integer from 20 to 400, n is an integer from 1 to 200, a is an integer greater than or equal to 4, and b is an integer from 1 to 4, where the monomer of the macro CTA polymer unit is in any sequence, and the styrene and macro CTA polymer unit are in any sequence.

[0052]

[0075] Macro-CTA polymer units may be quaternized with alkyl groups to modify the capture and killing efficiency of macro-CTA polymer units and / or to modify the responsiveness of macro-CTA polymer units (e.g., temperature, pH, salinity, light, and / or combinations thereof). In some embodiments, increasing the ratio of short-chain alkyl quaternization groups of 1 to 4 carbons to long-chain alkyl quaternization groups of 5 or more carbons, such as 5 to 20 carbons, increases the lower critical eutectic temperature (LCST) of macro-CTA polymer units in water. Unless expressly stated in the claims, we are not bound by any particular theory, but alkyl groups of 5 or more carbons, such as 5 to 20 carbons 2 It is thought that this can provide cell membrane penetration of the alkyl moiety into the hydrophobic portion of the cell membrane (such as the viral cell envelope, non-enveloped viral cell capsid, bacterial cell membrane, or fungal cell membrane). Unless explicitly stated in the claims, it is not bound by any particular theory, but it is thought that quaternary ammonium cations can provide interaction with the cell membrane surface (such as interaction with the phosphate portion of the phospholipid bilayer on the cell membrane surface). Unless explicitly stated in the claims, it is not bound by any particular theory, but it is thought that quaternary ammonium cations provide hydrophilicity to macro CTA polymer units, preventing the alkyl portion of the alkylamino group from being embedded in the polyalkene.

[0053]

[0076] In some embodiments, R 2The group is functionalized to contain a peptide sequence. The peptide sequence contains one or more amino acids and / or one or more amino acid components. Examples of amino acid peptide sequences include, but are not limited to, GRGD (Gly-Arg-Gly-Asp), RGD (Arg-Gly-Asp), and other peptide sequences. Examples of amino acid peptide sequences include glycoproteins such as vivonectin, fibronectin, and other glycoproteins. Examples of amino acid components include guanidine, butylammonium, imidazolium, and other groups. In one example, a peptide sequence using a peptide azide may be bonded to a quaternized amine group via a Cu(I)-catalyzed alkyne-azide (CuAAC) click reaction with an alkyne functional group. In another example, a peptide sequence using a peptide alkyne may be bonded to a quaternized amine group via a CuAAC click reaction with an azide functional group.

[0054]

[0077] In some embodiments, R 2 The group is functionalized, for example, through a CuAAC click reaction, to include a sugar sequence. The sugar sequence includes one or more monosaccharides, polysaccharides, and / or combinations thereof. Examples of sugars include fucose, glucose, mannose, galactose, GalNac, GlcNAc, sialic acid, other glycans, other amino sugars, other acid sugars, their derivatives, their isomers, their polysaccharides, and / or combinations thereof. In some embodiments, unless expressly stated in the claims, the functionalized sugar sequence R attached to the glycoprotein of an enveloped virus is not bound by any particular theory. 2 The substance is thought to interfere with the glycosylation of enveloped viruses, prevent them from attaching to host cells, and / or prevent them from entering host cells.

[0055]

[0078] In some embodiments, R 2The base is functionalized, for example, through a CuAAC click reaction, to include a protease (referring to any compound that breaks down peptides or amino acids). Examples of proteases include common peptide denaturants and asparagine, serine, threonine, or specific peptide denaturants that target their binding. In some embodiments, unless expressly stated in the claims, the invention is not bound to any particular theory, but includes functionalized proteases R bound to the glycoprotein of enveloped viruses. 2 The compound is thought to interfere with the glycosylation of enveloped viruses, bind to the capsid of non-enveloped viruses, prevent the attachment of viruses (enveloped or non-enveloped) to host cells, and / or prevent the entry of viruses (enveloped or non-enveloped) into host cells.

[0056]

[0079] In some embodiments, R 2 The base is functionalized, for example, through a CuAAC click reaction, to include a glycanase (referring to any compound that breaks down glycans). Examples of glycans include common glycan denaturants and specific glycan denaturants that target the binding of fucose, glucose, mannose, galactose, GalNac, GlcNAc, sialic acid, or their binding. In some embodiments, unless expressly stated in the claims, the functionalized glycanase R bound to the glycoprotein of an enveloped virus is not bound to any particular theory. 2 The substance is thought to interfere with the glycosylation of enveloped viruses, prevent them from attaching to host cells, and / or prevent them from entering host cells.

[0057]

[0080] Figure 4A shows a nanoworm 100 having a macro-CTA polymer having a quaternized amine group 140 containing an alkyne moiety, according to one embodiment. As shown in Figure 4B, a polymeric azide can react with the alkyne moiety in Figure 4A via a CuAAC click reaction to form a polymer 170 bonded to the macro-CTA polymer of the core 110 according to one embodiment. For example, the polymeric azide includes a peptide azide, a nucleic acid azide, a sugar azide, a protease azide, a glyconase azide, a polymer azide, or other polymeric azides, each forming a polymer 170 or other polymer containing a peptide sequence, a nucleic acid sequence, a sugar sequence, a protease, a glyconase, or a grafted polymer, respectively. In other embodiments, any polymer can be bonded to the quaternized amine group 140 of the macro-CTA polymer unit by any reaction scheme, for example, through the reaction of a macro-CTA polymer unit quaternized with an azide group reacting with a polymeric alkyne.

[0058]

[0081] Polymer 150 (as shown in Figure 2B) and / or Polymer 170 (as shown in Figure 4B) independently include, but are not limited to, grafted polymers having one or more properties, including temperature-responsive (LCST, UCST, or both) polymers, pH-responsive polymers, photoresponsive polymers, salt-responsive polymers, and / or combinations thereof. Grafted polymers include polymers obtained by any suitable polymerization method such as addition polymerization (including anionic and cationic polymerization), chain polymerization, free radical or living radical polymerization (including atom transfer polymerization or ATRP), metal-catalyzed polymerization, nitroxide polymerization, exchange chain transfer polymerization, RAFT, single electron transfer living radical polymerization or SET-LRP, condensation polymerization, and / or combinations thereof.

[0059]

[0082] Grafted polymers of polymer 150 (as shown in Figure 2B) and / or polymer 170 (as shown in Figure 4B) may be further functionalized. Grafted polymers may contain, or may be functionalized to contain, alkyl groups, carboxylic acids, alkynes, pyridines, dopamine, thiolactones, biotin, azides, peptide sequences, nucleic acid sequences, sugar sequences, proteases, glyconases, chromophore functional groups, other functional groups, derivatives thereof, and / or combinations thereof. For example, a grafted polymer may contain a quaternized and functionalized tertiary amine group. The quaternized amine group of the grafted polymer may be further functionalized to contain peptide sequences, nucleic acid sequences, sugar sequences, proteases, glyconases, and / or combinations thereof. The grafted polymer may contain two or more of the following functional groups to modify the capture and killing efficiency of nanoworms and / or to target common or specific microorganisms: alkyl, carboxylic acid, alkyne, pyridine, dopamine, thiolactone, biotin, azide, peptide sequence, nucleic acid sequence, sugar sequence, protease, glycanase, chromophore functional group, and other functional groups.

[0060]

[0083] The tertiary amine groups of grafted polymers may be quaternized with alkyl groups to modify the capture and killing efficiency of the grafted polymers and / or to modify the responsiveness of the grafted polymers (e.g., temperature, pH, salinity, light, and / or combinations thereof). In some embodiments, increasing the ratio of short-chain alkyl quaternization groups of 1 to 4 carbons to long-chain alkyl quaternization groups of 5 or more carbons, such as 5 to 20 carbons, increases the lower critical soluble temperature (LCST) of the grafted polymers in water. Unless expressly stated in the claims, we are not bound by any particular theory, but alkyl groups of 5 or more carbons, such as 5 to 20 carbons 2It is thought that this can provide cell membrane penetration of the alkyl portion into the hydrophobic portion of the cell membrane (such as the viral cell envelope, non-enveloped viral cell capsid, bacterial cell membrane, or fungal cell membrane). Unless explicitly stated in the claims, it is not bound by any particular theory, but it is thought that quaternary ammonium cations can provide interaction with the cell membrane surface (such as interaction with the phosphate portion of the phospholipid bilayer on the cell membrane surface). Unless explicitly stated in the claims, it is not bound by any particular theory, but it is thought that quaternary ammonium cations provide hydrophilicity to the grafted polymer, preventing the alkyl portion of the alkylamino group from becoming embedded in the core of the nanoworm.

[0061]

[0084] In some embodiments, the quaternized amine groups of the grafted polymer are functionalized to include a peptide sequence. The peptide sequence includes one or more amino acids and / or one or more amino acid components. Examples of amino acid peptide sequences include, but are not limited to, GRGD (Gly-Arg-Gly-Asp), RGD (Arg-Gly-Asp), and other peptide sequences. Examples of amino acid peptide sequences include glycoproteins such as vivonectin, fibronectin, and other glycoproteins. Examples of amino acid components include guanidine, butylammonium, imidazolium, and other groups. In one example, a peptide sequence using a peptide azide may be bonded to a quaternized amine group via a Cu(I)-catalyzed alkyne-azide (CuAAC) click reaction with a quaternized amine group containing an alkyne functional group. In another example, a peptide sequence using a peptide alkyne may be bonded to a quaternized amine group via a CuAAC click reaction with a quaternized amine group containing an azide functional group.

[0062]

[0085] In some embodiments, the quaternized amine groups of the grafted polymer are functionalized, for example, through a CuAAC click reaction, to include a sugar sequence. The sugar sequence includes one or more monosaccharides, polysaccharides, and / or combinations thereof. Examples of sugars include fucose, glucose, mannose, galactose, GalNac, GlcNAc, sialic acid, other glycans, other amino sugars, other acid sugars, their derivatives, their isomers, their polysaccharides, and / or combinations thereof. In some embodiments, although not bound by any particular theory unless expressly stated in the claims, a functionalized sugar sequence R attached to the glycoprotein of an enveloped virus is included. 2 The substance is thought to interfere with the glycosylation of enveloped viruses, prevent them from attaching to host cells, and / or prevent them from entering host cells.

[0063]

[0086] In some embodiments, the quaternized amine groups of the grafted polymer are functionalized, for example, through a CuAAC click reaction, to include proteases (referring to any compound that degrades peptides or amino acids). Examples of proteases include common peptide denaturants and asparagine, serine, threonine, or specific peptide denaturants that target their binding. In some embodiments, although not bound by any particular theory unless expressly stated in the claims, functionalized protease groups bound to the glycoprotein of an enveloped virus are thought to interfere with the glycosylation of the enveloped virus, bind to the capsid of a non-enveloped virus, interfere with the attachment of the virus (enveloped or non-enveloped) to the host cell, and / or interfere with the entry of the virus (enveloped or non-enveloped) into the host cell.

[0064]

[0087] In some embodiments, the quaternized amine groups of the grafted polymer are functionalized, for example, through a CuAAC click reaction, to include a glycanase (referring to any compound that breaks down glycans). Examples of glycans include common glycan denaturants and specific glycan denaturants that target the binding of fucose, glucose, mannose, galactose, GalNac, GlcNAc, sialic acid, or their binding. In some embodiments, although not bound by any particular theory unless expressly stated in the claims, a functionalized glycanase group bound to the glycoprotein of an enveloped virus is thought to interfere with the glycosylation of the enveloped virus, interfere with the attachment of the enveloped virus to a host cell, and / or interfere with the entry of the enveloped virus into a host cell.

[0065]

[0088] In one embodiment, a nanoworm comprising one or more sets of MacoCTA polymer units and one or more grafted polymers is general formula (VIII) (Alkene units) m (Macro CTA unit) n (Grafted polymer) (VIII) [In the formula, each of the macro CTAs is either non-functionalized or functionalized R 1 [Includes]. In one embodiment, the grafted polymer is the R of macro CTA. 1 It can be grafted onto the base. In another embodiment, the grafted polymer is grafted onto the quaternary amine group of the macro-CTA polymer unit. In yet another embodiment, the grafted polymer is grafted onto the R of the macro-CTA. 1 The group is grafted onto both the base and the quaternary amine group of the macro-CTA polymer unit. In some embodiments, m, n, and o are any independently selected positive integers. In some embodiments, m is an integer from 20 to 400, n is an integer from 1 to 200, and o is an integer from 1 to 10,000.

[0066]

[0089] In some embodiments of the nanoworms described herein, such as nanoworms of general formulas (IV)-(VII), the Z groups of macro-CTA polymer units as components of the RAFT agent may remain or be cleaved. In some embodiments, nanoworms containing Z groups of macro-CTA polymers as components of the RAFT agent may be further polymerized and / or crosslinked with other nanoworms.

[0067]

[0090] The nanoworms in Figures 1 to 21 are selected macro CTA polymer units, selected quaternization of tertiary amine groups of macro CTA polymer units, and R of macro CTA polymer units. 1 Selection of groups, quaternization of tertiary amine groups in macro CTA polymers 2 Selection of the group, including R for polymers 1 Selection of functionalization of the base, including R for polymers 2 Depending on the selection of the functionalization of the base, the selection of the grafted polymer incorporated into the nanoworm, the selection of the functionalization of the grafted polymer incorporated into the nanoworm, and / or combinations thereof, antimicrobial properties such as capture, killing, inactivating, degrading, degenerating, sterilizing, poisoning, and / or removing microorganisms may be obtained. Such selections may affect the responsiveness of the nanoworm to temperature, pH, salinity, and / or light.

[0068]

[0091] Macro CTA R 1 Based on the R of macro CTA 2 The virus-killing or inhibitory peptides or peptide mimeographs bound to the nanoworms and / or grafted polymers include, but are not limited to, the peptides listed in Tables 1-3. Table 1 includes peptides that inhibit viral adhesion and virus-cell membrane fusion. Table 2 includes peptides that interfere with the viral envelope. Table 3 includes peptides that inhibit viral replication. The peptides may target specific viruses or viruses across a broad spectrum. Other suitable peptides or peptide mimeographs may also be bound to the nanoworms to provide antimicrobial properties. TIFF0007872223000008.tif174170TIFF0007872223000009.tif77170TIFF0007872223000010.tif92170

[0069]

[0092] Figure 5 is a schematic cross-sectional view of a coating 400 comprising a plurality of nanoworms 100 on a substrate 420, such as one or more of the nanoworms from Figures 1 to 4 or Figures 6 to 21, according to one embodiment. An adhesion promoter 422 may optionally be used to increase the adhesion of the coating 400 on the substrate 420. The nanoworms 100 can be deposited on the substrate 420 by spraying, brushing, rolling, or other suitable deposition methods. The coating 400 provides antimicrobial properties to the substrate 420. The substrate 420 includes polymers, metals, cloths, glass, stone, ceramics, paper, other materials, and / or combinations thereof. The nanoworms 100 can be deposited as a dry powder. The nanoworms 100 can be deposited as a solution, such as diluted with water, organic solvents, inorganic solvents, and combinations thereof. Stored as a powder or as a water-diluted solution, the nanoworms 100 have a low or no risk of explosion.

[0070]

[0093] The coating 400 contains multiple identical nanoworms 100 or multiple different nanoworms 100. Some or all of the macro CTA polymer units 120 of the nanoworms may be exposed on the upper surface of the coating 400. The macro CTA polymer units 120 exposed on the upper surface of the coating 400 may be the same or different and may originate from multiple identical nanoworms or multiple different nanoworms 100. The macro CTA polymer units 120 exposed on the upper surface of the coating 400 may be individually hydrophobic or hydrophilic, or individually miscible or miscible with droplets, depending on the responsiveness of each individual macro CTA polymer unit 120, independently of the polyalkene properties of the nanoworms. For example, one or more of the macro CTA polymer units 120 may individually contain an LCST polymer, which is a class of water-soluble, thermoresponsive polymers that are miscible with droplets below the LCST of the macro CTA polymer unit 120. Above the LCST, the macro CTA polymer unit 120 is partially or entirely miscible with droplets. In other words, macro CTA polymer unit 120 is hydrophilic below its LCST and hydrophobic above its LCST.

[0071]

[0094] As shown in Figure 5, the droplet 450 may be miscible with the individual macro CTA polymer units 120 on the surface of the coating 400 at temperatures below the LCST of the individual macro CTA polymer units 120. The individual macro CTA polymer units 120, which are miscible with the droplet 450, may bind to or interact with the microorganisms 460 suspended in the droplet 450 or on the droplet 450, thereby providing antimicrobial properties to the substrate 420. The coating 400 may provide rapid or immediate antimicrobial properties to the microorganisms suspended in the droplet 450 or on the droplet 450 without waiting for the droplet 450 to evaporate. The coating 400 does not require a long evaporation time for the droplet in order to be effective in capturing or killing the microorganisms suspended in the droplet or on the droplet and preventing transmission. In contrast, antimicrobial hydrophobic coatings require a long evaporation time for the droplet to be effective. For example, a water droplet with an initial temperature of 20°C and an initial diameter of 3 mm will take approximately 111 minutes to completely evaporate at an ambient temperature of 24.8°C.

[0072]

[0095] In some embodiments, macro-CTA polymer units 120 can migrate between partially or entirely miscible and immiscible states accompanied by mucosal droplets. For example, macro-CTA polymer units 120 can migrate from below their LCST to above their LCST, or from above their LCST to below their LCST. Such migrations can result from changes in temperature, pH, salinity, other parameters, and / or combinations thereof. Migration of macro-CTA polymer units 120 between LCST conditions can impart compression, tension, or other mechanical forces to microorganisms, thereby providing antimicrobial properties.

[0073]

[0096] The coating 400 may be transparent, translucent, or opaque. The transparency of the coating can be determined by the packing density of the nanoworms 100 that produce the coating 400 and the selection of the polyalkene for the core 110 of the nanoworms 100. For example, a transparent coating allows for a light transmittance of at least about 80%, such as at least about 85%, at least about 90%, at least about 95%, or at least about 99% at one or more wavelengths from 380 nm to 740 nm. The light transmittance is determined by ASTM D 1003-00 (total transmittance), where the value indicated by T1 is for the transparent glass slide and the value indicated by T2 is for the nanoworm coating above the transparent glass slide. For example, a translucent coating allows for a light transmittance of at least about 30% to about 80% at one or more wavelengths from 380 nm to 740 nm. For example, an opaque coating allows for a light transmittance of less than 30% at one or more wavelengths from 380 nm to 740 nm. The opacity of the coating 400 can be determined by the packing density of the nanoworms 100 used to produce the coating 400, the selection of polyalkenes for the cores 110 of the nanoworms 100, and the addition of dyes and other additives.

[0074]

[0097] The coating 400 can be made permeable by controlling the packing density of the nanoworms 100 that make up the coating. The coating 400 can be made permeable by perforating, embossing, stretching, and / or calendering to form micropores. For example, the micropores may be in size from about 0.1 microns to about 10 microns.

[0075]

[0098] Figure 6 is a schematic cross-sectional view of a coating including a plurality of nanoworms 100 on a substrate 420 having a first set of macro CTA polymer units 120A and a second set of macro CTA polymer units 120B, where the first set of macro CTA polymer units 120A and the second set of macro CTA polymer units 120B have different LCSTs according to a certain embodiment. For example, both macro CTA polymer units 120A from the first set and macro CTA polymer units 120B from the second set can bind to microorganisms, while both macro CTA polymer units 120A and 120B have less than their LCST and are miscible with mucosal droplets 450. Subsequently, as shown in Figure 6, the macro CTA polymer units 120B are made partially or completely miscible with the droplets 450. Microorganisms 460 can be degraded by being bound to macro CTA polymer units 120A that are miscible with mucosal droplets 450, and by being partially or entirely linked to macro CTA polymer units 120B that are miscible with the droplets.

[0076]

[0099] A coating 400 containing multiple nanoworms, as shown in Figures 1 to 21, can be applied to any object. For example, a coating 400 containing multiple nanoworms 100 can be applied to a substrate 420 inside an aircraft (e.g., airplanes and helicopters) to provide antimicrobial properties to the aircraft's surface. The substrate 420 could be a tray table, headrest, seat back pocket, upper part of the seat back, seat armrest, seat, bathroom door lock, sink, toilet, in-flight magazine, safety card, overhead air vent, seat belt buckle, window shade, window, entertainment screen, interior wall, floor, pillow, blanket, and other surfaces of the aircraft. A coating 400 containing multiple nanoworms 100 can be applied to a substrate 420 inside a hospital to provide antimicrobial properties to the hospital's surface. The substrate 420 could be a bed, chair, table, counter, interior wall, floor, door handle, bathroom surface, and other surfaces of the hospital. A coating 400 containing multiple nanoworms 100 can be applied on top of a substrate 420, such as mounted on an exemplary substrate as described herein, in transport vehicles such as aircraft, spacecraft, buses, trains, subway cars, taxis, automobiles, ferries, boats, cruise ships, ride attractions, and other transport vehicles, to provide antimicrobial properties to the surface of the transport vehicle. The substrate 420 may be seats, handrails, doors, other similar objects as described herein, and other surfaces of the transport vehicle. A coating 400 containing multiple nanoworms 100 can be applied on top of a substrate 420 on the surface of a building such as an office building, school building, retail building, restaurant building, university building, daycare building, and other buildings, to provide antimicrobial properties to the surface of the building. The substrate 420 may be desks, chairs, tables, bathrooms, floors, interior walls, and other surfaces of the building. A coating 400 containing multiple nanoworms 100 can be applied on top of a substrate 420 on a sidewalk or people mover, to provide antimicrobial properties to the sidewalk or people mover. The substrate 420 may be a staircase, escalator, elevator, moving walkway, walkway handrail, elevator control button, and other walkway or people mover surface.A coating 400 containing multiple nanoworms 100 can be applied to a substrate 420 of food packaging to provide antimicrobial properties to the food packaging. The food packaging coating 400 may be at least partially transparent so that consumers can see the contents of the food packaging. The partially transparent coating allows for at least 30% light transmittance at one or more wavelengths from 380 nm to 740 nm. The light transmittance is determined by ASTM D 1003-00 (total transmittance), where the value indicated by T1 is for the transparent glass slide and the value indicated by T2 is for the nanoworm coating above the transparent glass slide. The food packaging coating 400 is breathable and can preserve the quality of the packaged food. The coating 400 containing multiple nanoworms 100 can be applied to a substrate 420 of an electronic device. For example, the electronic device may be a mobile device, headphones, keyboard, mouse, touchscreen, computer, or other electronic device. A coating 400 containing multiple nanoworms 100 can be applied to a wearable substrate 420, such as a substrate containing natural, synthetic, composite fibers, and fabrics, to provide antimicrobial properties to the wearable. Such wearables may be face masks, face shields, gloves, surgical gowns, hospital gowns, infant clothing, toddler clothing, or any wearable for which antimicrobial properties are desired. A coating 400 containing multiple nanoworms 100 can be applied to a medical device substrate 420. For example, medical devices include eye lenses, stents (e.g., coronary artery stents), artificial joints (e.g., knees), screws, pins, plates, rods, intrauterine devices, artificial discs, implants (e.g., breasts), prostheses, cardiac pacemakers, artificial hips, defibrillators, and other medical devices. A coating 400 containing multiple nanoworms 100 can be applied to a substrate 420 of a filter for any fluid. For example, the filter can filter air, blood, water, or other fluids to remove or kill microorganisms.

[0077]

[0100] A surface coated with nanoworms can provide antimicrobial activity to any amount of aqueous solution. For example, a surface coated with nanoworms can remove or kill microorganisms from blood in vivo and / or in vitro. For example, a surface coated with nanoworms can be used to filter donated blood in vitro to remove diseases such as coronavirus, HIV, hepatitis, syphilis, and other infectious diseases. For example, a surface coated with nanoworms may be used to treat a human patient by recirculating human patient-derived blood in vivo through a blood filter containing a nanoworm-coated surface to treat a viral or bacterial infection by removing or killing the virus or bacteria. For example, a surface coated with nanoworms may be used to treat a human patient by recirculating human patient-derived blood in vivo through a blood filter containing a nanoworm-coated surface to treat blood cancer, for example, by removing or killing cancerous leukemia, lymphoma, or myeloma cells.

[0078]

[0101] In some embodiments, the nanoworm composition can be applied to human skin as a solution or cream as a disinfectant for removing or killing microorganisms. In other embodiments, the nanoworm composition can be used as a drug applied topically, intravenously, or orally to the human body as a drug that targets common microorganisms, such as a general antiviral drug, or as a drug that targets specific microorganisms, such as a specific virus.

[0079] Clause

[0102] Clause 1. A nanoworm comprising multiple alkene units and R derived from a reversible addition-cleavage chain transfer agent. 1 A nanoworm comprising a first set of multiple macro CTA polymer units, each containing a group.

[0080]

[0103] Clause 2. The nanoworm according to Clause 1, wherein the first set of multiple macro CTA polymer units has a lower critical eutectic temperature (LCST) in water from -20°C to +100°C.

[0081]

[0104] Clause 3. The nanoworm according to Clause 1 or 2, wherein the first set of multiple macro CTA polymer units is configured to respond to temperature and to environmental conditions selected from the group consisting of pH, salinity, and light.

[0082]

[0105] Clause 4. R of the first set of macro CTA polymer units 1 A nanoworm according to any one of the clauses 1 to 3, wherein the group is a functional group selected from the group consisting of carboxylic acids, alkynes, pyridines, dopamine, thiolactones, biotin, azides, peptide sequences, sugar sequences, proteases, glycans, polymers, and combinations thereof.

[0083]

[0106] Clause 5. The nanoworm according to any one of Clauses 1 to 4, wherein the macro CTA polymer units of the first set comprise a quaternized amine.

[0084]

[0107] Clause 6. A nanoworm according to any one of Clauses 1 to 5, wherein the first set of macro-CTA polymer units is a functionalized quaternized amine selected from functional groups consisting of alkyl groups, carboxylic acids, alkynes, pyridines, dopamins, thiolactones, biotins, azides, peptide sequences, sugar sequences, proteases, glycans, polymers, and combinations thereof.

[0085]

[0108] Clause 7. A nanoworm according to any one of Clauses 1 to 6, wherein the first set of macro CTA polymer units comprises two or more sets of functionalized quaternary amines selected from functional groups consisting of alkyl groups, carboxylic acids, alkynes, pyridines, dopamins, thiolactones, biotins, azides, peptide sequences, sugar sequences, proteases, glycans, polymers, and combinations thereof.

[0086]

[0109] Clause 8. A nanoworm according to any one of Clauses 1 to 7, wherein the first set of macro CTA polymer units comprises a first set of functionalized quaternary amines of short-chain alkyl quaternary groups and a second set of functionalized quaternary amines of long-chain alkyl quaternary groups, wherein the short-chain alkyl group has 1 to 4 carbon atoms and the long-chain alkyl quaternary group has 5 or more carbon atoms.

[0087]

[0110] Clause 9. R derived from reversible addition-cleavage chain transfer agent 1 A nanoworm according to any one of the clauses 1 to 8, further comprising a second set of multiple macro CTA polymer units including a group, wherein the first set of multiple macro CTA polymer units is different from the second set of multiple macro CTA polymer units.

[0088]

[0111] Clause 10. The macro CTA polymer units of the first set are poly(N-isopropylacrylamide) (PNIPAM), poly(N,N-(dimethylamino)ethyl methacrylate) (F), poly(N-acetoxyethylacrylamide) (PNAEAA), poly(acryloylglycine ethyl ester) (PNAGEE), poly((ethylene glycol) methyl ether methacrylate) (PEGMEMA), poly((propylene glycol) methacrylate) (PPGMA), poly(N,N-dimethylacrylamide) (PDMA), A nanoworm according to any one of the claims 1 to 9, comprising a polymer selected from the group consisting of poly(N-decylacrylamide) (PDcA), poly(N,N-diethylacrylamide) (PDEA), poly(N-acryloylglycine) (PNAG), poly(N-acryloylglycine methyl ester) (PNAGME), poly(N-acryloylglycine ethyl ester) (PNAGEE), and poly(N-acryloylglycine propyl ester) (PNAGPE), polyacrylamide, polyacrylate, and copolymers thereof.

[0089]

[0112] Clause 11. A nanoworm as described in any one of Clauses 1 to 10, comprising at least a hydrophilic portion.

[0090]

[0113] Clause 12. A nanoworm as described in any one of Clauses 1 to 10, comprising a hydrophilic portion and a hydrophobic portion.

[0091]

[0114] Clause 13. A nanoworm according to any one of Clauses 1 to 12, further comprising a plurality of grafted polymers grafted onto at least a portion of a first set of a plurality of macro CTA polymer units.

[0092]

[0115] Clause 14. The nanoworm according to Clause 13, wherein the grafted polymer comprises a functionalized quaternized amine selected from functional groups consisting of alkyl, carboxylic acid, alkyne, pyridine, dopamine, thiolactone, biotin, azide, peptide sequence, sugar sequence, protease, glycanase, and combinations thereof.

[0093]

[0116] Clause 15. The nanoworm according to Clause 13 or 14, wherein the grafted polymer comprises two or more sets of functionalized quaternized amines selected from functional groups consisting of alkyl, carboxylic acid, alkyne, pyridine, dopamine, thiolactone, biotin, azide, peptide sequence, sugar sequence, protease, glycanase, and combinations thereof.

[0094]

[0117] Clause 16. The nanoworm according to any one of Clauses 13 to 15, wherein the grafted polymer comprises a first set of functionalized quaternary amines of short-chain alkyl quaternary groups and a second set of functionalized quaternary amines, wherein the short-chain alkyl group has 1 to 4 carbon atoms and the long-chain alkyl quaternary group has 5 or more carbon atoms.

[0095]

[0118] Clause 17. A nanoworm according to any one of Clauses 13 to 16, wherein the grafted polymer comprises a first set of functionalized quaternary amine groups containing a peptide sequence and a second set of functionalized quaternary amine groups containing a sugar sequence.

[0096]

[0119] Clause 18. The grafted polymer is R of the first set of macro CTA polymer units. 1A nanoworm as described in any one of clauses 13 to 17, grafted onto a base.

[0097]

[0120] Clause 19. A nanoworm according to any one of Clauses 13 to 18, wherein a grafted polymer is grafted onto a quaternary amine of a first set of macro-CTA polymer units.

[0098]

[0121] Clause 20. A first set of multiple grafted polymers is R of the macro CTA polymer units of the first set. 1 A nanoworm according to any one of the clauses 13 to 19, wherein a second set of multiple grafted polymers is grafted onto a quaternary amine of macro-CTA polymer units of the first set.

[0099]

[0122] Clause 21. The nanoworm according to any one of claims 13 to 20, wherein the grafted polymer is formed by a polymerization method selected from the group consisting of addition polymerization, chain polymerization, radical polymerization, metal-catalyzed polymerization, nitroxide polymerization, exchange chain transfer polymerization, RAFT, SET-LRP, condensation polymerization, and combinations thereof.

[0100]

[0123] A nanoworm according to any one of clauses 13 to 21, comprising 400 alkene units from clause 22.20, a first set of 1 to 200 macro-CTA polymer units, and 1 to 10,000 grafted polymers.

[0101]

[0124] Clause 23. A nanoworm according to any one of Clauses 1 to 22, comprising a peptide sequence capable of inhibiting viral attachment and viral-cell membrane fusion.

[0102]

[0125] Clause 24. A nanoworm according to any one of Clauses 1 to 23, comprising a peptide sequence capable of disrupting the viral envelope.

[0103]

[0126] Clause 25. A nanoworm according to any one of Clauses 1 to 24, comprising a peptide sequence capable of inhibiting viral replication.

[0104]

[0127] Clause 26. A nanoworm according to any one of Clauses 1 to 25, comprising a quaternized alkylquaternary ammonium cation capable of killing bacteria.

[0105]

[0128] Clause 27. A nanoworm as described in any one of Clauses 13 to 26, comprising at least a hydrophilic portion.

[0106]

[0129] Clause 28. A nanoworm as described in any one of Clauses 13 to 26, comprising a hydrophilic portion and a hydrophobic portion.

[0107]

[0130] Clause 29. A composition comprising a first set of a plurality of nanoworms and a second set of a plurality of nanoworms as described in any one of Clauses 1 to 28.

[0108]

[0131] Clause 30. A coating comprising one or a combination of the nanoworms described in any one of Clauses 1 to 28.

[0109]

[0132] Clause 31. The coating described in Clause 30, which is washable to replenish the antimicrobial properties of the nanoworms.

[0110]

[0133] Clause 32. The coating described in Clause 30 or 31, wherein the nanoworms are non-toxic.

[0111]

[0134] Clause 33. A coating as described in any one of Clauses 30 to 32, which is at least partially transparent.

[0112]

[0135] Clause 34. A coating according to any one of Clauses 30 to 33, including at least a hydrophilic portion.

[0113]

[0136] Clause 36. A coating according to any one of Clauses 30 to 33, comprising a hydrophilic portion and a hydrophobic portion.

[0114]

[0137] Clause 37. A transport body containing the coating described in any one of Clauses 30 to 35.

[0115]

[0138] Clause 38. An object containing a coating as described in any one of Clauses 30 to 35. [Examples]

[0116] material

[0139] Unless otherwise stated, all chemicals were used as received. Solvents included dichloromethane (DCM, Aldrich AR grade), DMSO (Aldrich, 99.9%), n-hexane (Emsure, ACS reagent), chloroform (Emsure, ACS reagent), acetone (ChemSupply, AR grade), petroleum refined (BR 40-60℃, Univar, AR grade), toluene (EMSURE, ACS reagent, ISO, Reag. Ph Eur), ethyl acetate (ChemSupply, AR grade), and N,N-dimethylacetamide (Aldrich, >99%). Other materials included activated basic alumina (Aldrich: Brockmann). I, standard grade, approximately 150 mesh, 58A), magnesium sulfate (anhydrous, ultrapure), sodium chloride (ChemSupply, AR grade), Milli-Q water (Biolab, 18.2 MΩm), sodium dodecyl sulfate (SDS, Aldrich, 99%), N,N′-dicyclohexylcarbodiimide (DCC, Aldrich, 99%), 4-(dimethylamino)pyridine (DMAP, Merck, 99%), 1-butanethiol (Aldrich, 99%), propargyl alcohol (Aldrich, 99%), lithium chloride (Aldrich, 99%), tripotassium phosphate (Aldrich, ≥98%), sodium bicarbonate (Aldrich Aldrich, 99.5%, hydrochloric acid (36%, Ajax, AR grade), sulfuric acid (Aldrich, 98%), hydrogen peroxide (Aldrich, 30 wt.% in water, ACS reagent), carbon disulfide (Aldrich, >99.9%), 2-bromo-2-methylpropionic acid (Aldrich, 98%), methyl-2-bromopropionate (MBP, Aldrich, 98%), iodooctane (Aldrich, 98%), iodomethane (Aldrich, 99%, containing copper as a stabilizer), copper(II) sulfate (Aldrich, 99%), L-ascorbic acid (Aldrich, 99%), poly(ethyleneimine) solution (PEI, Aldrich, 50 wt.% in water, Mn It contained 1800 (Mw 2000), GRGD (Gly-Arg-Gly-Asp) azide (Auspep, 97%), and a glass surface.Styrene (STY, Aldrich, >99%) and N,N-(dimethylamino)ethyl methacrylate (DMAEMA, Aldrich, 98%) were passed through a basic alumina column to remove all inhibitors. N-isopropylacrylamide (NIPAM, Aldrich, 97%) was recrystallized from n-hexane / toluene (9 / 1, v / v). Azobisisobutyronitrile (AIBN, Riedel-de Haen) was recrystallized twice from methanol before use. Ethyl α-bromoisobutyrate (EBiB, Aldrich, 98%).

[0117] Analysis method

[0140] nuclear magnetic resonance (NMR) All NMR spectra were performed using a Bruker DRX 400 MHz spectrometer with external locking (CDCl3 or DMSO-d6).

[0118]

[0141] Size exclusion chromatography (SEC) and triple detection size exclusion chromatography (TD-SEC) The molecular weight distribution of the polymers was analyzed using a Polymer Laboratories GPC50 Plus equipped with a differential refractive index detector. The absolute molecular weight of the polymers was determined using a Polymer Laboratories GPC50 Plus equipped with a dual-angle laser light scattering detector, a viscometer, and a differential refractive index detector. High-performance liquid chromatography (HPLC) grade N,N-dimethylacetamide (DMAc, containing 0.03 wt% LiCl) was used as the eluent at a flow rate of 1.0 mL / min. Separation was achieved using two PLGel Mixed B (7.8 × 300 mm) SEC columns connected in series and maintained at a constant temperature of 50°C. The triple detection system was calibrated using a 5 mg / mL 110 K polystyrene (PSTY) standard. Samples of known concentrations were freshly prepared in DMAc + 0.03 wt% LiCl and passed through a 0.45 μm PTFE syringe filter before injection. The absolute molecular weight and dn / dc values ​​were determined using Polymer Laboratories Multi Cirrus software based on quantitative mass recovery.

[0119]

[0142] Dynamic light scattering (DLS) Dynamic light scattering was performed using a Malvern Zetasizer Nano Series 3000HS running DTS software with a 4mW He-Ne laser at 0.633nm. The analysis was performed at an angle of 173° and a temperature of 25°C. Number-mean hydrodynamic particle diameter (D h ) and the multivariance index (PDI) DLS ) was measured. PDI DLS This explains the width of the particle size distribution and was calculated from the cumulant analysis of the intensity autocorrelation function measured by DLS. It is also related to the standard deviation of the assumed Gaussian distribution (i.e., PDI). DLS =σ 2 / ZD 2 [In the formula, σ is the standard deviation and ZD is the mean size of Z.]

[0120]

[0143] Lower critical cosolution temperature (LCST) To determine the lower critical eutectic temperature (LCST) of macro-CTA, macro-CTA was dissolved in 10 mg / mL Milli-Q water in an ice bath. The solution was then filtered directly into a DLS cuvette using a 0.45 μm cellulose syringe filter. The polymer solution was cooled to 5°C, and the cuvette was placed in the DLS instrument. Measurements were performed by slowly raising the temperature from 5°C to 70°C at a ramp rate of 2°C / min, controlled by standard operating procedure software.

[0121]

[0144] Upper critical solution temperature (LCST) To determine the lower critical eutectic temperature (LCST) of macro-CTA, macro-CTA was dissolved in Milli-Q water in a 70°C water bath. The solution was then filtered into a DLS cuvette using a 0.45 μm (micrometer) cellulose syringe filter. The cuvette was placed in a DLS instrument. Measurements were performed by slowly cooling the polymer solution from 70°C to less than 1°C.

[0122] RAFT agent

[0145] Ester-functionalized RAFT agentsEster-functionalized RAFT agents (ester RAFT agents) of methyl 2-(butylthiocarbonothiothio)propanoate (MCEBTTC) RAFT agent were synthesized according to the following reaction scheme (I). TIFF0007872223000011.tif82170

[0123]

[0146] Carboxylic acid-functionalized RAFT agent Carboxylic acid-functionalized RAFT agents (acid RAFT agents) were synthesized according to reaction scheme (II). TIFF0007872223000012.tif82170

[0124]

[0147] Alkyne-functionalized RAFT agent Alkyne-functionalized RAFT agents (alkyne RAFT agents) were synthesized according to reaction scheme (III). TIFF0007872223000013.tif80170

[0125] Synthesis of poly(NIPAM) macroCTA using ester RAFT agents

[0148] Macro CTAs of NIPAM polymers were synthesized according to reaction scheme (IV). TIFF0007872223000014.tif113170

[0126]

[0149] The concentration ratio of NIPAM:RAFT(MCEBTTC):AIBN was 44:1:0.1, and the ratio of DMSO to NIPAM was 2 / 1 (v / w). NIPAM (4.31 g, 3.81 × 10⁻² mol), MCEBTTC (0.219 g, 8.68 × 10⁻⁴ mol), and AIBN (14.2 mg, 8.65 × 10⁻⁵ mol) were dissolved in DMSO (8.6 mL). The mixture was deoxygenated by purging with argon for 40 minutes and heated at 70°C for 18 hours. Polymerization was then interrupted by cooling in an ice bath at 0°C and exposure to air. The solution was diluted with chloroform (200 mL) and washed five times with 40 mL of Milli-Q water. The chloroform was then dried over anhydrous MgSO₄, filtered, and the volume was reduced by rotary evaporation. The polymer was recovered by precipitation in a large excess of diethyl ether (400 mL), isolated by filtration, and then dried under vacuum at room temperature for 24 hours to obtain a yellow powder product. The polymer product was then processed into macro(PNIPAM). 44 This is called )-A (macro CTA-A).

[0127] Synthesis of poly(NIPAM / DMAEMA) macroCTA using alkyne RAFT agents

[0150] Macro-CTA copolymers of NIPAM and DMAEMA were synthesized according to reaction scheme (V). TIFF0007872223000015.tif107170

[0128]

[0151] The concentrations of NIPAM:DMAEMA:RAFT:AIBN were 50:30:1:0.15, and the ratio of DMSO to NIPAM and DMAEMA was 1.1 / 1 (v / w). NIPAM (1.99 g, 1.76 × 10⁻² mol), DMAEMA (1.66 g, 1.05 × 10⁻² mol), RAFT alkyne (0.102 g, 3.51 × 10⁻⁴ mol), and AIBN (8.6 mg, 5.27 × 10⁻⁵ mol) were dissolved in DMSO (4 mL). The mixture was deoxygenated by purging with argon for 40 minutes, heated to 70°C, and polymerized for 17 hours. The reaction was then interrupted by cooling in an ice bath at 0°C and exposure to air. The solution was then diluted with 4 mL of chloroform, precipitated in a large amount of excess petroleum extract (250 mL), and isolated by centrifugation. The dissolution and precipitation cycle was repeated three times. The product was then dissolved in Milli-Q water and freeze-dried to recover a yellow powder. The polymer product was then processed using macro(P(NIPAM). 50 -co-DMAEMA 35 This is called ))-B (macro CTA-B).

[0129] Synthesis of poly(NIPAM / DMAEMA) macro-CTA using ester RAFT agents

[0152] Macro CTAs of NIPAM and DMAEMA copolymers were synthesized according to reaction scheme (VI). TIFF0007872223000016.tif114170

[0130]

[0153] The concentrations of NIPAM:DMAEMA:RAFT:AIBN were 50:30:1:0.15, and the ratio of DMSO to NIPAM and DMAEMA was 1.1 / 1 (v / w). NIPAM (2 g, 1.76 × 10⁻² mol), DMAEMA (1.66 g, 1.05 × 10⁻² mol), RAFT MCEBTTC (0.089 g, 3.53 × 10⁻⁴ mol), and AIBN (8.7 mg, 5.29 × 10⁻⁵ mol) were dissolved in DMSO (4 mL). The mixture was deoxygenated by purging with argon for 40 minutes, then heated at 70°C and polymerized for 16 hours. The reaction was then interrupted by cooling in an ice bath at 0°C and exposure to air. The solution was then diluted with 4 mL of chloroform and precipitated in a large amount of excess petroleum extract (250 mL), and then isolated by centrifugation. The dissolution and precipitation cycle was repeated three times. The product was then dissolved in Milli-Q water and freeze-dried to recover a yellow powder. The polymer product was then processed using macro(P(NIPAM). 50 -co-DMAEMA 32 This is called ))-C (macro CTA-C).

[0131] Classification of poly(NIPAM / DMAEMA) macro CTA into quaternary classes.

[0154] Macro CTAs of NIPAM and DMAEMA copolymers were synthesized according to reaction scheme (VII). TIFF0007872223000017.tif179170

[0132]

[0155] Alkyne RAFT polymerizer P(NIPAM50-co-DMAEMA35) (macro(P(NIPAM50-co-DMAEMA35)-B, 50 mg, mw=11451)) was dissolved in DCM (1.2 mL). Then, iodooctane (240.13 g / mol, 1.33 g / cm³) was added to the polymer solution, and the mixture was shaken at 23°C for 8 hours. Next, iodomethane (141.94 g / mol, 2.28 g / cm³) was added, and the mixture was shaken further at 23°C for 11 hours. The polymer solution was dialyzed against acetone (3 × 500 mL), followed by dialyzed against Milli-Q water (3 × 500 mL) (MWCO 3500). The sample was lyophilized to obtain a white powder as the product. Different ratios of iodooctane and iodomethane were used according to Table 4. TIFF0007872223000018.tif46170

[0133] Temperature-Directed Morphological Change (TDMT) Method for Generating Nanoworms

[0156] Nanoworms were generated using styrene emulsion polymerization with macro(PNIPAM44)-A and macro(P(NIPAM50-co-DMAEMA35))-B. In a Schlenk tube, macro(PNIPAM44)-A (40 wt.%, 70 mg, 1.3 × 10⁻⁵ mol), macro(P(NIPAM50-co-DMAEMA35))-B (60 wt.%, 105 mg, 9.2 × 10⁻⁶ mol), and SDS (7.25 mg, 2.5 × 10⁻⁵ mol) were dissolved in cold Milli-Q water (3.25 mL). The mixture was deoxygenated by purging with argon for 20 minutes. AIBN (0.37 mg, 2.2 × 10⁻⁶ mol) was dissolved in styrene (0.1304 g, 1.3 × 10⁻³ mol), and the solution was injected into a macro CTA. This was then purged with argon in an ice bath for an additional 5 minutes before heating to 70°C. Polymerization was interrupted after 4 hours by exposing the reaction mixture to air at 70°C.

[0134]

[0157] 70 °C latex (1 mL) was mixed with 15 - 20 μL of toluene, cooled to 23 °C, and left standing at 23 °C for 1 hour. Then, the solution was gradually cooled to 10 °C over 10 minutes and left standing at 10 °C for 20 hours. The nanostructure was characterized by TEM to confirm the formation of worm-like nanostructures, and then freeze-dried to obtain a white powder.

[0135] Protocol for post-modification of alkyne PDMAEMA nanoworms (40 WT% macro CTA-A and 60 WT% macro CTA-B)

[0158] Nanoworm Example 1 (Alkyne P (DMAEMA) Nanoworm) : Alkyne-terminated poly(DMAEMA) nanoworms containing poly(NIPAM) and alkyne-terminated poly(NIPAM-co-DMAEMA) were generated as follows. In a Schlenk tube, macro(PNIPAM44)-A (70 mg, 1.3×10−5 mol), macro(P(NIPAM50-co-DMAEMA35))-B (105 mg, 9.2×10−6 mol), and SDS (7.25 mg, 2.5×10−5 mol) were dissolved in cold Milli-Q water (3.25 mL). The mixture was deoxygenated by purging with argon for 20 minutes. AIBN (0.37 mg, 2.2×10−6 mol) was dissolved in styrene (0.1304 g, 1.3×10−3 mol), and the solution was injected into the macro CTA mixture, which was then purged with argon in an ice bath for an additional 5 minutes before heating to 70 °C. The reaction was terminated after 4 hours by exposing the reactants to air at 70 °C. 70 °C latex (1 mL) was mixed with 20 microliters (μL) of toluene, cooled to 23 °C, and left standing at 23 °C for 1 hour. Then, the solution was gradually cooled to 10 °C over 10 minutes and left standing at 10 °C for 20 hours. The nanostructure was characterized by TEM to confirm the formation of worm-like nanostructures, and then freeze-dried to obtain a white powder.

[0136]

[0159] Nanoworm Example 2 (Peptide P (DMAEMA) Nanoworm): By functionalizing the alkyne-terminated poly(NIPAM-co-DMAEMA) of nanoworm Example 1 to be terminated with the GRGD peptide, peptide-terminated poly(DMAEMA) nanoworms were generated as follows. GRGD-azide (1 equivalent) and alkyne worm (nanoworm Example 1, 1 equivalent) were dispersed in a mixture of Milli-Q water and DMSO (10% v.) (2 mL). The mixture was deoxygenated by purging with argon for 40 minutes. CuSO4 (3 equivalents) was dissolved in 0.6 mL of Milli-Q water / DMSO (10% v.) and purged with Ar for 20 minutes. Ascorbic acid (7 equivalents) was dissolved in 0.6 mL of Milli-Q water / DMSO (10% v.) and purged with Ar for 20 minutes. The ascorbic acid solution was injected into the suspension of worm and GRGD-azide using a degassed syringe, followed by the injection of the CuSO4 solution. After stirring at 23 °C for 19 hours, the reaction was interrupted by exposing to air, and the suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO). The resulting solution was lyophilized to obtain a powder.

[0137]

[0160] Nanoworm Example 3 (Alkyne P (DMAEMA) nanoworm quaternized with iodomethane) : The alkyne poly(DMAEMA) nanoworms quaternized with iodomethane were generated as follows by quaternizing the alkyne-terminated poly(NIPAM-co-DMAEMA) of nanoworm Example 1 with a methyl group. Nanoworm Example 1 (VB-B10-R67A, 20 mg, MW 12619) was resuspended in Milli-Q water (90%) / DMSO (10%) (total volume 1 mL). Then, 1.6 μL of iodomethane (141.94 g / mol, 2.28 g / cm 3 ) was added to the nanoworm suspension, and shaking was continued at 23 °C for 19 hours. The suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO). The resulting solution was lyophilized to obtain a powder.

[0138]

[0161] Nanoworm Example 4 (Alkyne P (DMAEMA) nanoworms quaternized with iodoctane (10%) and iodomethane (90%))Alkyne poly(DMAEMA) nanoworms quaternized with iodomethane were produced by quaternizing the alkyne-terminated poly(NIPAM-co-DMAEMA) of Nanoworm Example 1 with methyl and octyl groups, as follows: The DMAEMA group of the macro CTA of Nanoworm 1 was quaternized with methyl and octyl groups. Nanoworm Example 1 (VB-B10-R67A, 20 mg, MW 12619) was resuspended in Milli-Q water (90%) / DMSO (10%) (total volume 1 mL). Then, 0.4 μL of iodooctane (240.13 g / mol, 1.33 g / cm3) was added using the stock solution, and the mixture was shaken at 23°C for 8 hours. Subsequently, 1.3 μL of iodomethane (141.94 g / mol, 2.28 g / cm³) was added, and the mixture was shaken at 23°C for 11 hours. The suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0139]

[0162] Nanoworm Example 5 (Peptide P(DMAEMA) nanoworm quaternized with iodomethane) : Peptide poly(DMAEMA) nanoworms quaternized with iodomethane were produced by quaternizing the peptide terminal poly(DMAEMA) of Nanoworm Example 2 with a methyl group, as follows: Nanoworm Example 2 (MW 12824) was resuspended in Milli-Q water (90%) / DMSO (10%) (total volume 1 mL). Then, 0.92 μL of iodomethane (141.94 g / mol, 2.28 g / cm3) was added to the nanoworm suspension and shaken continuously at 23°C for 19 hours. The suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO). The obtained solution was freeze-dried to obtain a powder.

[0140]

[0163] Nanoworm Example 6 (Quaternized (peptide P(DMAEMA) nanoworm) with iodoctane (10%) and iodomethane (90%)):Peptide poly(DMAEMA) nanoworms quaternized with iodooctane (10%) and iodomethane (90%) were produced by quaternizing the peptide terminal poly(DMAEMA) of Nanoworm Example 2 with methyl and octyl groups, as follows: Nanoworm 1, functionalized with GRGD peptide (VB-B10-R69, mg, MW 12824), was resuspended in Milli-Q water (90%) / DMSO (10%) (total volume 1 mL). Then, 0.24 μL of iodooctane (240.13 g / mol, 1.33 g / cm3) was added using the stock solution, and the mixture was shaken at 23°C for 8 hours. Then, 1 μL of iodomethane (141.94 g / mol, 2.28 g / cm3) was added, and the mixture was shaken at 23°C for 11 hours. The suspension was dialyzed in Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0141]

[0164] Nanoworm Example 7 (Alkyne P (DMAEMA) nanoworm quaternized with iodoctane (10%) and iodomethane (90%)) Alkyne poly(DMAEMA) nanoworms quaternized with iodooctane (10%) and iodomethane (90%) were produced by quaternizing the alkyne-terminated poly(NIPAM-co-DMAEMA) of Nanoworm Example 1 with methyl and octyl groups, as follows: Nanoworm Example 1 (40 mg, MW 11994) was resuspended in Milli-Q water (85%) / DMSO (15%) (total volume 1.5 mL). Then, 0.1 equivalents of iodooctane (0.864057 μL, 240.13 g / mol, 1.33 g / cm3) were added and the mixture was shaken at 23°C for 8 hours. Then, 0.9 equivalents of iodomethane (2.681392 μL, 141.94 g / mol, 2.28 g / cm3) were added and the mixture was shaken at 23°C for 11 hours. The suspension was dialyzed in Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0142]

[0165] Nanoworm Example 8 (Alkyne P (DMAEMA) nanoworm quaternized with iodoctane (30%) and iodomethane (70%))Alkyne poly(DMAEMA) nanoworms quaternized with iodooctane (30%) and iodomethane (70%) were produced by quaternizing the alkyne-terminated poly(NIPAM-co-DMAEMA) of Nanoworm Example 1 with methyl and octyl groups, as follows: Nanoworm Example 1 (40 mg, MW 11994) was resuspended in Milli-Q water (85%) / DMSO (15%) (total volume 1.5 mL). Then, 0.3 equivalents of iodooctane (2.592172 μL, 240.13 g / mol, 1.33 g / cm3) were added and the mixture was shaken at 23°C for 8 hours. Then, 0.7 equivalents of iodomethane (2.085527 μL, 141.94 g / mol, 2.28 g / cm3) were added and the mixture was shaken at 23°C for 11 hours. The suspension was dialyzed in Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0143]

[0166] Nanoworm Example 9 (Alkyne P (DMAEMA) nanoworm quaternized with iodoctane (50%) and iodomethane (50%)) Alkyne poly(DMAEMA) nanoworms quaternized with iodooctane (50%) and iodomethane (50%) were produced by quaternizing the alkyne-terminated poly(NIPAM-co-DMAEMA) of Nanoworm Example 1 with methyl and octyl groups, as follows: Nanoworm Example 1 (40 mg, MW 11994) was resuspended in Milli-Q water (85%) / DMSO (15%) (total volume 1.5 mL). Then, 0.5 equivalents of iodooctane (4.320287 μL, 240.13 g / mol, 1.33 g / cm3) were added and the mixture was shaken at 23°C for 8 hours. Then, 0.5 equivalents of iodooctane (1.489662 μL, 141.94 g / mol, 2.28 g / cm3) were added and the mixture was shaken at 23°C for 11 hours. The suspension was dialyzed in Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0144]

[0167] Nanoworm Example 10 (Alkyne P (DMAEMA) nanoworm quaternized with iodoctane (70%) and iodomethane (30%))Alkyne poly(DMAEMA) nanoworms quaternized with iodooctane (70%) and iodomethane (30%) were produced by quaternizing the alkyne-terminated poly(NIPAM-co-DMAEMA) of Nanoworm Example 1 with methyl and octyl groups, as follows: Nanoworm Example 1 (40 mg, MW 11994) was resuspended in Milli-Q water (85%) / DMSO (15%) (total volume 1.5 mL). Then, 0.7 equivalents of iodooctane (6.048402 μL, 240.13 g / mol, 1.33 g / cm3) were added and the mixture was shaken at 23°C for 8 hours. Then, 0.3 equivalents of iodomethane (0.893797 μL, 141.94 g / mol, 2.28 g / cm3) were added and the mixture was shaken at 23°C for 11 hours. The suspension was dialyzed in Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0145]

[0168] Nanoworm Example 11 (Alkyne P (DMAEMA) nanoworm quaternized with iodoctane (90%) and iodomethane (10%)) Alkyne poly(DMAEMA) nanoworms quaternized with iodooctane (90%) and iodomethane (10%) were produced by quaternizing the alkyne-terminated poly(NIPAM-co-DMAEMA) of Nanoworm Example 1 with methyl and octyl groups, as follows: Nanoworm Example 1 (40 mg, MW 11994) was resuspended in Milli-Q water (85%) / DMSO (15%) (total volume 1.5 mL). Then, 0.9 equivalents of iodooctane (7.776517 μL, 240.13 g / mol, 1.33 g / cm3) were added and the mixture was shaken at 23°C for 8 hours. Then, 0.1 equivalents of iodomethane (0.297932 μL, 141.94 g / mol, 2.28 g / cm3) were added and the mixture was shaken at 23°C for 11 hours. The suspension was dialyzed in Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0146]

[0169] Nanoworm Example 12 (Quaternized (peptide P(DMAEMA) nanoworm) with iodoctane (90%) and iodomethane (10%)):Peptide poly(DMAEMA) nanoworms quaternized with iodooctane (90%) and iodomethane (10%) were functionalized with GRGD peptide at the end of the alkyne-terminated poly(NIPAM-co-DMAEMA) of nanoworm Example 7, which was to be terminated, as follows: GRGD-azide (1 equivalent) and alkyne worm (nanoworm Example 7, 1 equivalent) were dispersed in a degassed mixture of Milli-Q water and DMSO (10% v.) (1.4 mL). Degassed aqueous ascorbic acid (7 equivalents) was injected into the suspension of worm and GRGD-azide using a degassed syringe, followed by the injection of aqueous CuSO4 (3 equivalents). After stirring at 23°C for 19 hours, the reaction was interrupted by exposure to air, and the suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0147]

[0170] Nanoworm Example 13 (Quaternized (peptide P(DMAEMA) nanoworm) with iodoctane (30%) and iodomethane (70%)) :Peptide poly(DMAEMA) nanoworms quaternized with iodooctane (30%) and iodomethane (70%) were functionalized with GRGD peptide to the alkyne-terminated poly(NIPAM-co-DMAEMA) of nanoworm Example 8, which was to be terminated, as follows: GRGD-azide (1 equivalent) and alkyne worm (nanoworm Example 8, 1 equivalent) were dispersed in a degassed mixture of Milli-Q water and DMSO (10% v.) (1.4 mL). Degassed aqueous ascorbic acid (7 equivalents) was injected into the suspension of worm and GRGD-azide using a degassed syringe, followed by the injection of aqueous CuSO4 (3 equivalents). After stirring at 23°C for 19 hours, the reaction was interrupted by exposure to air, and the suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0148]

[0171] Nanoworm Example 14 (Quaternized (peptide P(DMAEMA) nanoworm) with iodoctane (50%) and iodomethane (50%)):Peptide poly(DMAEMA) nanoworms quaternized with iodooctane (50%) and iodomethane (50%) were functionalized with GRGD peptide to the alkyne-terminated poly(NIPAM-co-DMAEMA) of nanoworm Example 9, which was to be terminated, as follows: GRGD-azide (1 equivalent) and alkyne worm (nanoworm Example 9, 1 equivalent) were dispersed in a degassed mixture of Milli-Q water and DMSO (10% v.) (1.4 mL). Degassed aqueous ascorbic acid (7 equivalents) was injected into the suspension of worm and GRGD-azide using a degassed syringe, followed by the injection of aqueous CuSO4 (3 equivalents). After stirring at 23°C for 19 hours, the reaction was interrupted by exposure to air, and the suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0149]

[0172] Nanoworm Example 15 (Quaternized (peptide P(DMAEMA) nanoworm) with iodoctane (70%) and iodomethane (30%)) :Peptide poly(DMAEMA) nanoworms quaternized with iodooctane (70%) and iodomethane (30%) were prepared by functionalizing the alkyne-terminated poly(NIPAM-co-DMAEMA) of nanoworm Example 10, which was to be terminated, with GRGD peptide as follows: GRGD-azide (1 equivalent) and alkyne worm (nanoworm Example 10, 1 equivalent) were dispersed in a degassed mixture of Milli-Q water and DMSO (10% v.) (1.4 mL). Degassed aqueous ascorbic acid (7 equivalents) was injected into the suspension of worm and GRGD-azide using a degassed syringe, followed by the injection of aqueous CuSO4 (3 equivalents). After stirring at 23°C for 19 hours, the reaction was interrupted by exposure to air, and the suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0150]

[0173] Nanoworm Example 16 (Quaternized (Peptide P(DMAEMA) Nanoworm) with Iodooctane (90%) and Iodomethane (10%)):Peptide poly(DMAEMA) nanoworms quaternized with iodooctane (90%) and iodomethane (10%) were functionalized with GRGD peptide to the alkyne-terminated poly(NIPAM-co-DMAEMA) of nanoworm Example 11, which was to be terminated, as follows: GRGD-azide (1 equivalent) and alkyne worm (nanoworm Example 11, 1 equivalent) were dispersed in a degassed mixture of Milli-Q water and DMSO (10% v.) (1.4 mL). Degassed aqueous ascorbic acid (7 equivalents) was injected into the suspension of worm and GRGD-azide using a degassed syringe, followed by the injection of aqueous CuSO4 (3 equivalents). After stirring at 23°C for 19 hours, the reaction was interrupted by exposure to air, and the suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO). The resulting solution was freeze-dried to obtain a powder.

[0151] Temperature-responsive macro CTA

[0174] Three pH-responsive and temperature-responsive macro-CTAs were synthesized with different amounts of NIPAM and DMAEMA prepared by reversible addition-cleavage chain transfer (RAFT) polymerization. All macro-CTAs contained trithioester (or RAFT) terminal groups, allowing for further polymerization with other monomers to form copolymers, which could then be transferred in situ to nanoworms. The three macro-CTAs were characterized using size exclusion chromatography (SEC) and NMR, and are listed in Table 5. For further copper-catalyzed alkyne-azide addition-cyclization (CuAAC) "click" reactions, macro-CTA-B consisted of alkyne functional groups. Within the relatively narrow molecular weight distribution, polymerization was all well controlled.

[0152]

[0175] The lower critical eutectic temperature (LCST) of three macro CTAs was determined in water using a DLS instrument. The LCST was characterized by the increase in LCST shape size at a specific temperature. All three macro CTAs shown in Table 5 have an LCST close to 30°C, which provides an excellent LCST for generating nanoworms that offer antimicrobial properties. TIFF0007872223000019.tif61170TIFF0007872223000020.tif60170

[0153]

[0176] The conversion rate (%) was calculated by comparing the incorporation of the polymer and the residual monomer from 1H NMR. The total conversion rate (total) of the polymer was calculated as follows: total conversion = [[(conversion (NIPAM) × mNIPAM) + (conversion (DMAEMA) × mDMAEMA)] / (mNIPAM + mDMAEMA)] × 100. Mn (theoretical) was calculated from the procedure described in J. Am. Chem. Soc., 2015, 137(50), 15652 - 15655. Mn (1H NMR) was determined from 1H NMR. Using polystyrene as a calibration standard and eDMAc + 0.03 wt.% LiCl as an eluent, Mn (SEC Abs) was calculated. The LCST is defined here as the lowest temperature at which the macro CTA is partially or entirely insoluble in the specified liquid medium.

[0154]

[0177] Figure 7 is a graph showing an example of determining the LCST of macro CTA in water at different weight fractions and different pHs according to one embodiment. LCST profiles of macro CTA-C (0.05 wt.%) at different wt% in sodium chloride solution (71 mg / mL). The pH was adjusted to 6.592 with HCl (0.14 M) (curve a). Then, additional macro CTA-C was added to obtain concentrations of 3 mg / mL (0.3 wt%, curve b), 10.5 mg / mL (1.04 wt%, curve c), or 57.7 mg / mL (5.5 wt%, curve d). The pH of the solution was changed to approximately 9 - 10. The LCST at each weight fraction of the polymer was measured.

[0155]

[0178] Different weight fractions and pH levels simulate the change in polymer properties (or polymer structure from coil to sphere) from water-soluble to water-insoluble when water (or mucous) droplets adhere to a surface coated with nanoworms, and the surface properties of the coating are primarily governed by the macro CTA polymer units of the nanoworms. The aqueous solution contains a large amount of salt (71 mg / mL) to simulate the large amount of salt found in bodily fluids such as saliva, urine, sweat, and other bodily fluids. With low weight fraction polymer (0.05 wt%) at pH=6.5, no LCST was observed up to 70°C, suggesting that the droplet initially changes the polymer-coated surface to be highly water-soluble, resulting in rapid wetting of the droplet across the entire surface and thus providing high microbial capture efficiency. When the weight fraction and pH were increased to 0.3 wt% at pH=8-9, the LCST was approximately 19.5°C. When the weight fraction and pH were increased to 1.04 wt% at pH = 9-10, the LCST was approximately 14°C. When the weight fraction and pH were further increased to 5.5 wt% at a pH of approximately 9.3, the LCST was approximately 13°C.

[0156]

[0179] This demonstrated that the initial droplets on the surface reduce the concentration of macro-CTA polymer units, but as the droplets evaporate, the weight fraction of the macro-CTA polymer increases. As the weight fraction increases, the buffering capacity of the macro-CTA polymer units in alkali DMAEMA increases, which increases the pH. At a certain weight fraction and pH, the polymer surface reverts to its initial water-insoluble state.

[0157]

[0180] Figure 8 is a graph showing an example of incorporating cationic and hydrophobic moieties into macro CTA according to a certain embodiment. The LCST profiles of macro CTA-B after post-modification with different iodine compounds (Milli-Q 10 mg / mL in water) are as follows: (a) No quaternization (LCST 27°C); (b) 100% iodomethane (LCST 55°C); (c) iodomethane (90%) and iodocatane (10%) (LCST 40°C); (d) iodomethane (70%) and iodocatane (30%) (LCST 35°C); (e) iodomethane (50%) and iodocatane (50%) (LCST 35°C); (f) iodomethane (30%) and iodocatane (70%) (LCST 35°C); (g) iodomethane (10%) and iodocatane (90%) (LCST 29°C).

[0158]

[0181] The cationic and hydrophobic portions are highly effective in killing bacteria. As shown in Scheme 1A, iodomethane and iodoctane were reacted with amines in DMAEMA to produce quaternary cationic amines with different ratios of methane and octane groups on the surface of nanoworms. The LCST was determined for macro CTAs with various ratios, as shown in Figure 8. As the octane ratio increased, the LCST decreased from approximately 55°C (100% methane) to 30°C (90% octane). This demonstrated that the LCST of nanoworms can be fine-tuned to specific environmental conditions.

[0159]

[0182] Figure 9 shows examples of LCST profiles of macro-CTA-B (10 mg / mL) after quaternation with iodomethane (50%) and iodoctane (50%) at different pH levels in Milli-Q water or sodium chloride (NaCl) solution, according to one embodiment: (a) Milli-Q water (pH=8.28; LCST=35°C); (b) Milli-Q water (pH=6.05; LCST=45°C); (c) Milli-Q water (pH=9.60; LCST=33°C); (d) NaCl solution (71 mg / mL, pH=6.19; no LCST was observed in the temperature range investigated); (e) NaCl solution (71 mg / mL, pH=9.53, LCST=45°C).

[0160]

[0183] To test the changes in the polymer structure of quaternized macro-CTA-B containing 50% methane and 50% octane, the LCST was determined by varying both pH and salt content. Curve (a) represents a solution of Q macro-CTA-B dissolved in Milli-Q water, where the pH was determined to be 8.28 and the LCST was 35°C. Adjusting the pH to 6.05 using HCl increased the LCST to 45°C, while raising the pH to 9.6 using NaOH decreased the LCST to 33°C. Adding a salt (NaCl) close to the amount of salt found in artificial body fluids resulted in no LCST being observed at pH 6.19, but raising the pH to 9.53 resulted in an LCST being observed at 45°C. This data indicates that salt increases the LCST, while increasing pH decreases it.

[0161] TDMT method for generating nanoworms

[0184] The molecular weights of the macro-CTA and polystyrene (PSTY) copolymer were determined by SEC and NMR. Table 6 shows that the copolymer is well controlled with 45 to 48 units of polystyrene after three polymerization cycles. The initial nanoparticle sizes at 70°C, before transformation into nanoworms at low temperatures, ranged from 163 to 215 nm, indicating a very narrow particle size distribution (PDIDLS << 0.1). By gradually cooling these emulsions from 70°C to 10°C, nanoworms were reproducibly generated, as confirmed by TEM microscopy. The nanoworms shown in Scheme 1A consisted of alkyne-terminated groups that could further bind to a wide range of molecules and polymers. TEM images of nanoworms were prepared by TDMT from latex spheres obtained by RAFT emulsion polymerization of styrene using macro-CTA-A (40 wt%) and macro-CTA-B (60 wt%). Three replication polymerizations were performed to investigate reproducibility. TIFF0007872223000021.tif34170

[0162]

[0185] Post-modification of nanoworms with either or both iodine compounds (i.e., iodomethane and iodooctane) and / or the integrin-conjugated peptide GRGD.

[0163]

[0186] The above nanoworms were modified with methane and octane groups in various ratios, with or without GRGD (see Table 7). The zeta potential of all nanoworms (i.e., nanoworm examples) was generally above +30mV, with the only exception being the nanoworm example representing the initial nanoworm before modification. Post-modification did not alter the nanoworm structure. TIFF0007872223000022.tif163170

[0164] Surface wettability

[0187] Nanoworm deposition on glass surfaces by spin coating Prior to spin coating, the glass surface was washed with a "piranha" solution (H2SO4 + H2O2), followed by five washes in a 70% ethanol solution. The surface was prepared by spin coating a nanoworm aqueous solution onto the glass surface. 100 microliters (μL) of nanoworm aqueous solution (5 mg / mL) was added to the dry glass surface and spin-coated at 2000 rpm for 60 seconds with a spin acceleration of 400 rpm / second. The spin-coated surface was then dried at ambient temperature for 10 hours.

[0165]

[0188] Surface wetting test using concentrated salt solution at 23°C A 10 μL salt solution (71 mg / mL sodium chloride in Milli-Q water, pH 6.5) was placed on a test surface at 23°C, and a video of the droplet behavior on the surface was recorded for 1 minute. This salt solution has a salt content and pH similar to artificial saliva.

[0166]

[0189] To determine how the pH of this salt solution affects surface wetting, four salt solutions (71 mg / mL sodium chloride in Milli-Q water, 0.04 mg / mL orange II sodium salt) were prepared at different pH values ​​of 6.25, 7.60, 8.35, and 9.30. Then, 5 μL of each of these salt solutions at 23°C were placed on a test surface at 23°C, and the behavior of droplets on the surface was observed.

[0167]

[0190] Surface wetting test using artificial saliva solution at 50°C A 10 μL salt solution (71 mg / mL sodium chloride in Milli-Q water, pH 6.5) at 50°C was placed on a test surface at 23°C, and the behavior of the liquid on the surface was observed.

[0168]

[0191] Surface wetting test using Milli-Q water (0.04 mg / mL Orange II sodium chloride) at different pH levels at 23°C. 5 μL of Milli-Q water at pH 5.95 was placed on a nanoworm-coated surface at 23°C, and the behavior of droplets on the surface was observed. The same procedure was repeated on nanoworm-coated surfaces using Milli-Q water with pH 7.29, 8.75, or 9.96.

[0169]

[0192] Surface wetting test using Milli-Q water (0.04 mg / mL Orange II sodium chloride) at different pH levels at 37°C. 5 μL of Milli-Q water at 37°C and pH 5.95 was placed on a test surface at 37°C, and the behavior of droplets on the nanoworm-coated surface was observed. The same procedure was repeated using Milli-Q water at pH 7.29, 8.75, or 9.96.

[0170]

[0193] Results of surface wettingThe glass surface was first cleaned with piranha solution and then spin-coated with nanoworms as shown in Table 7. Droplets of water, with or without NaCl (similar to the salts found in artificial bodily fluids), at 23°C or 50°C, were dropped onto the nanoworm-coated slide (23°C). The slide coated with Nanoworm Example 1 was first tested with NaCl droplets at either 23°C or 50°C. Regardless of the droplet temperature, the droplet rapidly wetted the entire surface in just a few seconds, and a photograph was taken after 60 seconds. This suggests that the droplet temperature on the surface rapidly equilibrates to the ambient temperature (23°C in this case). When a droplet was placed on the surface coated with or without NaCl (71 mg / mL), the entire surface was quickly wetted. When the droplet temperature and surface temperature were 37°C in pure water, no observable wetting occurred after 60 seconds. This demonstrated that the surface is water-insoluble. When slides were coated with Nanoworm Example 3 and Nanoworm Example 4, the LCST decreased as the octane group content in the nanoworm increased, and therefore the surface became water-insoluble under these conditions. In summary, the ability of surfaces designed to be highly water-soluble or water-insoluble under various environmental conditions was demonstrated. Rapid wetting demonstrates that under many types of conditions (similar to artificial body fluid conditions), microorganisms have a greater chance of being captured compared to current antimicrobial surfaces.

[0171] Antibacterial properties of nanoworms

[0194] The following protocol was used to test nanoworm-coated surfaces against E. coli. E. coli was cultured at OD(600nm)=0.2, 100 microliters (μL) were taken out and centrifuged at 8000×g, 4°C, for 5 minutes. The pellet was resuspended in 900 μL of PBS and completely resuspended by vortexing at maximum speed for 3 minutes. The above solution was diluted 1000-fold. 10 μL of the diluted E. coli was taken out and added to the polymer-coated coverslip surface. Incubated at RT for 3 hours. The coverslip was transferred to a tube containing 300 μL of PBS and incubated at room temperature for 5 minutes. The tube was vortexed for 5 seconds at low power (e.g., speed 1-2). 300 μL of PBS was transferred to a new Eppendorf 1.5 mL tube. Briefly vortex a 1.5 mL tube and transfer 60 μL of the solution onto an LB agar plate (containing kanamycin, 50 μg / mL). Incubate the plate overnight at 37°C. Count the clones on the plate.

[0172]

[0195] Figure 10 shows an example of the antimicrobial activity against E. coli on a glass surface coated with nanoworms in one embodiment. Each surface was replicated three times. The surfaces are as follows: Untreated corresponds to a bare glass surface; PEI corresponds to a glass surface coated with PEI (Mn 1800); No. 1 corresponds to Nanoworm Example 1; No. 2 corresponds to a glass surface coated with Nanoworm Example 2 (GRGDPDMAEMA worm); No. 4 corresponds to a glass surface coated with Nanoworm Example 4; No. 8 corresponds to a glass surface coated with Nanoworm Example 8; No. 9 corresponds to a glass surface coated with Nanoworm Example 9; No. 12 corresponds to a glass surface coated with Nanoworm Example 12; No. 13 corresponds to a glass surface coated with Nanoworm Example 13; No. 14 corresponds to a glass surface coated with Nanoworm Example 14.

[0173]

[0196] Glass slips were coated with various nanoworms shown in Table 7, and their antimicrobial (E. coli) activity was tested. To determine the killing efficiency of the nanoworm surface, a control surface of PEI was used and set to approximately 40% killing efficiency. PEI is known to kill E. coli rapidly and effectively. Nanoworm Examples 1, 2, 4, 8, 9, 12, 13, and 14 tested killed more bacteria than PEI. A surface coated with Nanoworm Example 1 (having a low positive zeta potential) achieved a killing efficiency of approximately 45%. When GRGD was applied (Nanoworm Example 2), the killing efficiency increased to approximately 75% compared to Nanoworm Example 1. Nanoworms functionalized with methyl and octyl groups showed increased killing efficiency compared to Nanoworm Example 1. Nanoworm Examples 12, 13, and 14, which possessed both GRGD groups and alkyl groups, resulted in near 100% elimination efficiency.

[0174] Nanoworms based on alkyne-γ-thiolactone PDMAEMA

[0197] Synthesis of alkyne-γ-thiolactone PDMAEMA nanoworms

[0198] In a Schlenk tube, non-functional PNIPAM 44 -S(C=S)SC4H9(30wt.%, 0.8077g, 1.51×10 -4 mol), γ-thiolactone P (NIPAM) 43 -co-DMA 20 )-S(C=S)SC4H 9( 10 wt.%, 0.2692 g, 3.74 × 10 -5 mol), alkynes (P(NIPAM) 50 -co-DMAEMA 35 )-S(C=S)SC4H 9( 60 wt.%, 1.6154 g, 1.55 × 10 -4 (mol), and SDS (0.1115g, 3.87×10) -4 0.202 mL of styrene (1.76 × 10⁻⁶ mol) was dissolved in degassed cold Milli-Q water (50 mL). -3The mixture was deoxygenated by adding AIBN (0.0056 g, 3.43 × 10) via syringe and purging with argon for 1 hour. -5 mol) to styrene (2 mL, 1.818 g, 1.75 × 10) -2 The solution was dissolved in (mol) and injected into a macro CTA mixture, which was then purged with argon in an ice bath for a further 25 minutes before being heated to 70°C. Polymerization was interrupted after 5 hours by exposing the reactants to air at 70°C.

[0175]

[0199] Figure 14 shows a schematic diagram of the synthesized alkyne-γ-thiolactone PDMAEMA nanoworm 1400. The polyalkene units and macro-CTA polymer units form the core 1410. Functionality R at the ends of the macro-CTA 1 The base extends from the core and forms macro CTA hairs 1420 located at arbitrary positions along the core. For example, alkyne (P(NIPAM) 50 -co-DMAEMA 35 )-S(C=S)SC4H9 macro CTA alkyne R 1 The group extends from core 1410. For example, γ-thiolactone P (NIPAM) 43 -co-DMA 20 )-S(C=S)SC4H9 macro CTA γ-thiolactone R 1 The base extends from core 1410.

[0176] Quaternization of alkyne-γ-thiolactone PDMAEMA nanoworms

[0200] Quaternization with 10% equivalent iodoctane

[0201] Figure 14 shows alkyne-γ-thiolactone PDMAEMA nanoworms (DMAEMA group, 1.6g, 1.83 × 10⁻⁶). -3 The iodoctane (33 μL, 0.044 g, 1.83 × 10) was redispersed in 25 mL of Milli-Q water. -4Iodooctane (mol) was dissolved in 4.41 mL of DMSO. The solution containing iodooctane in DMSO was added to an alkyne-γ-thiolactone PDMAEMA nanoworm dispersion, and the reaction mixture was shaken at 23°C for 19 hours. Subsequently, the reaction mixture was dialyzed against a 0.01 M sodium thiosulfate solution (3500 MWCO, 3 × 1 L, alternating every 4 hours), followed by dialyzed against Milli-Q water (3500 MWCO, 3 × 1 L, alternating every 4 hours). The reaction mixture was freeze-dried, and the product was isolated as a white powder.

[0177]

[0202] Figure 15A shows a schematic diagram of synthetic alkyne-γ-thiolactone PDMAEMA nanoworm 1500 quaternized with 10% equivalent iodooctane. It is estimated that approximately 10% of the tertiary amine groups in the PDMAEMA polymer unit of core 1510 are quaternized with octyl groups.

[0178]

[0203] Quaternization with 10% equivalent iodooctane and 90% equivalent iodomethane

[0204] Figure 14 shows alkyne-γ-thiolactone PDMAEMA nanoworms (DMAEMA group, 1.96 g, 2.05 × 10⁻⁶). -3 The iodoctane (37 μL, 0.049 g, 2.05 × 10) was redispersed in 25 mL of Milli-Q water. -4 Iodooctane (115 μL, 0.261 g, 1.84 × 10⁻¹⁶) was dissolved in 4.38 mL of DMSO. The solution containing iodooctane in DMSO was added to an alkyne-γ-thiolactone PDMAEMA nanoworm dispersion, and the reaction mixture was shaken at 23°C for 9 hours. Subsequently, iodomethane (115 μL, 0.261 g, 1.84 × 10⁻¹⁶) was added. -3 The reaction mixture was further reacted at 23°C for 15 hours after adding (mol) of the reaction solution. The reaction mixture was then dialyzed against a 0.01 M sodium thiosulfate solution (3500 MWCO, 3 × 1 L, alternating every 4 hours), followed by dialyzed against Milli-Q water (3500 MWCO, 3 × 1 L, alternating every 4 hours). The reaction mixture was freeze-dried to isolate the product as a white powder.

[0179]

[0205] Figure 15B shows a schematic diagram of synthetic alkyne-γ-thiolactone PDMAEMA nanoworm 1500, which has been quaternized with 10% equivalent iodooctane and 90% equivalent iodomethane. It is estimated that approximately 10% of the tertiary amine groups of the PDMAEMA polymer units of core 1510 are quaternized with octyl groups, and approximately 90% of the tertiary amine groups of the PDMAEMA polymer units of core 1510 are quaternized with methyl groups. Nanoworm 1600 corresponds to Nanoworm Example 17(P3).

[0180]

[0206] Quaternization with 100% equivalent propargyl bromide

[0207] The alkyne-γ-thiolactone PDMAEMA nanoworm (1.7084 g of DMAEMA groups, 1.76 × 10⁻³ mol) shown in Figure 14 was redispersed in 20 mL of Milli-Q water. Propargyl bromide (80 wt.% in toluene, 196 μL, 0.209 g, 1.76 × 10⁻³ mol) was dissolved in 3.53 mL of DMSO. The solution containing propargyl bromide in DMSO was added to the alkyne-γ-thiolactone PDMAEMA nanoworm dispersion, and the reaction mixture was shaken at 23°C for 19 hours. Subsequently, the reaction mixture was dialyzed against Milli-Q water (3500 MWCO, 5 × 1 L, changed every 4 hours). The reaction mixture was freeze-dried, and the product was isolated as a white powder.

[0181]

[0208] Figure 15C shows a schematic diagram of synthetic alkyne-γ-thiolactone PDMAEMA nanoworm 1500, quaternized with 100% equivalent propargyl bromide. It is estimated that approximately 100% of the tertiary amine groups in the PDMAEMA polymer unit of core 1510 are quaternized with propargyl groups.

[0182] Conjugation of guanidine azide to quaternary alkyne-γ-thiolactone PDMAEMA nanoworms via CuAAC Conjugation of quaternized alkyne-γ-thiolactone PDMAEMA nanoworms containing approximately 10% of tertiary amine groups quaternized by octyl groups.

[0210] A quaternated alkyne-γ-thiolactone PDMAEMA nanoworm (1.485 g of alkyne groups, 5.55 × 10⁻⁵ mol) having approximately 10% of tertiary amine groups quaternized by octyl groups, as shown in Figure 15A, was dispersed in 20 mL of a degassed solution (85% / 15%, vv.) of Milli-Q water and DMSO. A solution containing guanidine azide (0.0095 g, 6.66 × 10⁻⁵ mol) in 1 mL of a degassed solution (85% / 15%, vv.) of Milli-Q water and DMSO was injected into the nanoworm dispersion via a degassed syringe. A degassed solution (85% / 15%, vv.) of Milli-Q water and DMSO containing 5 mL of ascorbic acid (0.0684 g, 3.88 × 10⁻⁴ mol) was injected into a dispersion of nanoworms and guanidine azide via a degassed syringe. Subsequently, a degassed solution (85% / 15%, vv.) of Milli-Q water and DMSO containing 5 mL of CuSO₄ (0.0265 g, 1.66 × 10⁻⁴ mol) was injected into the reaction mixture via a degassed syringe. The reaction was carried out under an argon atmosphere at 23°C for 19 hours. The reaction was interrupted by exposure to air, and the suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO, 9 × 1 L, changed every 4 hours). The reaction solution was freeze-dried to obtain the product as a white powder.

[0183]

[0211] Figure 16A shows a schematic diagram of a conjugated guanidine azide synthesized on a quaternized alkyne-γ-thiolactone PDMAEMA nanoworm having approximately 10% of tertiary amine groups quaternized by octyl groups. Nanoworm 1600 corresponds to Nanoworm Example 18 (P6).

[0184]

[0212] Conjugation of quaternized alkyne-γ-thiolactone PDMAEMA nanoworms having approximately 10% tertiary amine groups quaternized with octyl groups and approximately 90% tertiary amines quaternized with methyl groups.

[0185]

[0213] Figure 15B shows quaternary alkyne-γ-thiolactone PDMAEMA nanoworms (0.25g of alkyne group, 8.47 × 10⁻⁶). -6 The guanidine azide (0.0014 g, 1.02 × 10⁻⁶) was dispersed in 10 mL of degassed Milli-Q water and DMSO solution (85% / 15%, vv.). -5A solution containing (mol) Milli-Q water and DMSO in 1 mL of degassed solution (85% / 15%, vv.) was injected into the nanoworm dispersion via a degassed syringe. 2 mL of ascorbic acid (0.0104 g, 5.93 × 10⁻⁶) was added. -5 A degassed solution of Milli-Q water and DMSO (85% / 15%, vv.) containing (mol) was injected into a dispersion of nanoworms and guanidine azide via a degassed syringe. Subsequently, 2 mL of CuSO4 (0.0041 g, 2.54 × 10) was added. -5 A degassed solution (85% / 15%, vv.) of Milli-Q water and DMSO containing 1 mol was injected into the reaction mixture via a degassed syringe. The reaction was carried out under an argon atmosphere at 23°C for 19 hours. The reaction was interrupted by exposure to air, and the suspension was dialyzed against Milli-Q water for 36 hours (3500 MWCO, 9 × 1 L, changed every 4 hours). The reaction solution was freeze-dried to obtain the product as a white powder.

[0186]

[0214] Figure 16B shows a schematic diagram of a conjugated guanidine azide synthesized on a quaternized alkyne-γ-thiolactone PDMAEMA nanoworm having approximately 10% quaternized tertiary amine groups with octyl groups and approximately 90% quaternized tertiary amines with methyl groups. Nanoworm 1600 corresponds to Nanoworm Example 19 (P10).

[0187] Nanoworms based on P(NIPAM-co-DMAEMA)-N3

[0215] P(NIPAM) by single-electron transfer living radical polymerization 55 -co-DMAEMA 48 ) synthesis

[0216] Macro CTAs of NIPAM and DMAEMA copolymers were synthesized by single-electron transfer living radical polymerization according to reaction scheme (VIII). TIFF0007872223000023.tif133170

[0188]

[0217] In a 10 mL Schlenk tube equipped with a magnetic stirrer, add Cu(II)Br 2(0.0197g, 8.84 × 10 -5 (mol) and NaBH4 (0.0033g, 8.84 × 10⁻⁶) -5 (mol) was added. The flask was sealed with a rubber septum and purged with Ar for 30 minutes. Me6TREN (24 μL, 0.02 g, 8.84 × 10) was added to a 5 mL glass vial. -5 Add (mol) and Milli-Q water (2.564 mL), seal the vial, and purge the solution with Ar for 30 minutes. Transfer this solution to a Cu(II)Br2 / NaBH4 Schlenk tube using a cannula, place it in an ice bath, and here Cu II The reduction was allowed to proceed for 30 minutes. NIPAM (1g, 8.84 × 10 -3 (mol) and DMAEMA (0.893 mL, 0.833 g, 5.30 × 10) -3 (mol) and initiator EBiB (16 μL, 0.0215 g, 1.10 × 10) -4 Another mixture (mol) was dissolved in isopropanol (2.564 mL) in a 5 mL glass vial, sealed, purged with argon at 0°C for 30 minutes, and then transferred to a polymerization Schlenk tube via cannula. Polymerization was carried out at 0°C for 90 minutes.

[0189]

[0218] P(NIPAM 55 -co-DMAEMA 48 )-N 3 synthesis

[0219] P(NIPAM 55 -co-DMAEMA 48 Macro CTA of )-N3 was synthesized according to reaction scheme (IX). TIFF0007872223000024.tif143170

[0190]

[0220] 2 mL of water containing 20 equivalents of NaN3 was added to the polymerization mixture of reaction scheme (VIII), and the mixture was heated to 25°C and then stirred overnight. The reaction mixture was then dialyzed against acetone (3 × 1 L, alternating every 3 hours). The dialyzed solution was passed through activated basic alumina to remove the copper salts. The solvent was removed by rotary evaporation, 5 mL of Milli-Q water was added, and the residue was dried and frozen to isolate the product as a white powder.

[0191]

[0221] P(NIPAM) in 10% equivalent iodoctane 55 -co-DMAEMA 48 )-N 3 Classification to four levels

[0222] Macro CTAs of NIPAM and DMAEMA copolymers were synthesized according to reaction scheme (X). TIFF0007872223000025.tif122170

[0192]

[0223] P(NIPAM 55 -co-DMAEMA 48 )-N3(DMAEMA group, 7.7178g, 2.64×10 -2 (mol) was dissolved in 50 mL of DCM. Then, iodooctane (0.476 mL, 0.634 g, 2.64 × 10) -3 A mol of thiosulfate was added, and the reaction was carried out at 23°C for 19 hours. The reaction mixture was then dialyzed against a 0.01 M sodium thiosulfate solution (3500 MWCO, 3 × 1 L, alternating every 4 hours), followed by dialyzed against Milli-Q water (3500 MWCO, 3 × 1 L, alternating every 4 hours). The reaction mixture was freeze-dried, and the product was isolated as a white powder. Quaternization (NIPAM) was performed with the octyl group according to reaction scheme (X). 55 -co-DMAEMA 48 )-N3 was formed. Here, X was approximately 55, Y(1-Z%) was approximately 43.2, and Y(Z%) was approximately 4.8.

[0193] Grafting of quaternized P(NIPAM-CO-DMAEMA)-N3 onto alkyne-γ-thiolactone PDMAEMA nanoworms

[0224] Quaternized P(NIPAM 55 -co-DMAEMA 48 )-N 3 CuAAC-mediated grafting of alkyne-γ-thiolactone PDMAEMA nanoworms

[0225] Quaternized P(NIPAM) containing 10% equivalents of iodooctane (reaction product of scheme X) 55 -co-DMAEMA 48 )-N3(5.484g, 3.75×10 -4 The mol of propargyl group (alkyne group) was dissolved in 90 mL of Milli-Q water / DMSO mixture (85% / 15%, vv.). Then, this solution was dissolved in the propargyl group (alkyne group) shown in Figure 15C (0.5975 g, 6.25 × 10⁻⁶). -4 Quaternally quaternated alkyne-γ-thiolactone PDMAEMA nanoworms were quaternized (mol) and the mixture was purged with argon for 3 hours. 10 mL of ascorbic acid (0.7706 g, 4.38 × 10⁶) -3 A degassed solution of Milli-Q water (85% / 15%, vv.) containing 100ml of CuSO4 (0.2993g, 1.88×10) and DMSO was injected into the reaction mixture via a degassed syringe. Then, 10 mL of CuSO4 (0.2993g, 1.88×10) was added. -3 A degassed solution (85% / 15%, vv.) of Milli-Q water and DMSO containing 1 mol was injected into the reaction mixture via a degassed syringe. The reaction was carried out under an argon atmosphere at 23°C for 19 hours. The reaction was interrupted by exposure to air, and the suspension was dialyzed against 0.01 M EDTA disodium salt (3500 MWCO, 3 × 1 L, alternating every 4 hours), followed by dialyzed against Milli-Q water for 36 hours (3500 MWCO, 9 × 1 L, alternating every 4 hours). The resulting solution was freeze-dried to obtain the product as a bright blue powder.

[0194]

[0226] Figure 17 shows quaternized P(NIPAM) grafted onto alkyne-γ-thiolactone PDMAEMA nanoworm 1700A. 55 -co-DMAEMA 48A schematic diagram is shown. The obtained nanoworm 1700A contains a core 1710 containing an alkyne-γ-thiolactone PDMAEMA nanoworm. Quaternized P(NIPAM) 55 -co-DMAEMA 48 )-N3 portion 1730 is the alkyne R of alkyne-γ-thiolactone PDMAEMA nanoworm 1 It is grafted onto the base. Quaternary P(NIPAM) 55 -co-DMAEMA 48 The 1740 portion of )-N3 is grafted onto the alkyne group of the propargyl group of the quaternized alkyne-γ-thiolactone PDMAEMA nanoworm.

[0195]

[0227] Quaternization of grafted PDMAEMA nanoworms with 30% propargyl bromide, and conjugation of guanidine azide to quaternized grafted alkyne PDMAEMA nanoworms.

[0228] Figure 17A: Grafted PDMAEMA nanoworm (2.2759 g of DMAEMA group, 5.94 × 10⁶) -3 The mol of propargyl bromide was redispersed in 80 mL of Milli-Q water. Propargyl bromide (80 wt.% in toluene, 198 μL, 0.212 g, 1.78 × 10⁻¹⁶) -3 A mol (mol) of propargyl bromide was dissolved in 14.12 mL of DMSO. The solution containing propargyl bromide in DMSO was added to a grafted PDMAEMA nanoworm dispersion, and the reaction mixture was shaken at 23°C for 19 hours. The reaction mixture was then dialyzed against Milli-Q water (3500 MWCO, 6 × 1 L, changed every 4 hours). The reaction mixture was freeze-dried, and the product was isolated as a bright blue powder.

[0196]

[0229] Figure 17B shows the obtained P(NIPAM) grafted onto the alkyne-γ-thiolactone PDMAEMA nanoworm 1700B. 55 -co-DMAEMA 48 A schematic diagram showing ) is shown. P(NIPAM 55 -co-DMAEMA 48 Parts 1730 and 1740 of ) were quaternized with propargyl and octyl groups. Approximately 30% of the remaining tertiary amine group Y(1-Z%) was quaternized with propargyl groups.

[0197]

[0230] Figure 17B shows quaternary grafted alkyne-γ-thiolactone PDMAEMA nanoworm (1.5067 g of alkyne group, 1.15 × 10⁻⁶). -3 The guanidine azide (0.163 g, 1.15 × 10⁻⁶ mol) was dispersed in 18 mL of degassed solution (85% / 15%, vv.) of Milli-Q water and DMSO. -3 A solution containing 10 mL of ascorbic acid (1.412 g, 8.02 × 10) in a 2 mL degassed solution of Milli-Q water and DMSO (85% / 15%, vv.) was injected into the nanoworm dispersion via a degassed syringe. -3 A degassed solution of Milli-Q water and DMSO (85% / 15%, vv.) containing 10 ml of CuSO4 (0.5484 g, 3.44 × 10) was injected into a dispersion of nanoworms and guanidine azide via a degassed syringe. Subsequently, 10 mL of CuSO4 (0.5484 g, 3.44 × 10) was added. -3 A degassed solution (85% / 15%, vv.) of Milli-Q water and DMSO containing 1 mol was injected into the reaction mixture via a degassed syringe. The reaction was carried out under an argon atmosphere at 23°C for 19 hours. The reaction was interrupted by exposure to air, and the suspension was dialyzed against 0.01 M EDTA disodium salt (3500 MWCO, 3 × 1 L, alternating every 4 hours), followed by dialyzed against Milli-Q water for 36 hours (3500 MWCO, 9 × 1 L, alternating every 4 hours). The resulting solution was freeze-dried to obtain the product as a light green powder.

[0198]

[0231] Figure 17C shows a schematic diagram illustrating the conjugation resulting from guanidine azide to quaternized grafted alkyne PDMAEMA nanoworm 1700C. Guanidine azide is used in portions 1730 and 1740 of P(NIPAM). 55 -co-DMAEMA 48 It was conjugated to the propargyl group of ). Nanoworm 1700C corresponds to Nanoworm Example 20 (P8).

[0199]

[0232] Quaternization of grafted PDMAEMA nanoworms with 50% propargyl bromide, and conjugation of guanidine azide to quaternized grafted alkyne PDMAEMA nanoworms.

[0233] By quaternizing approximately 50% of the remaining tertiary amine group Y(1-Z%) with propargyl groups, and by conjugating guanidine azide to propargyl quaternization groups, quaternized grafted alkyne PDMAEMA nanoworms similar to those in Figure 17C were formed. These formed nanoworms correspond to Nanoworm Example 21(P9).

[0200]

[0234] Conjugation of guanidine azide and polygalactose azide into quaternary grafted alkyne PDMAEMA nanoworms

[0235] Figure 17B shows quaternary grafted alkyne-γ-thiolactone PDMAEMA nanoworm (0.5g of alkyne group, 3.80 × 10⁻⁶). -4 The guanidine azide (0.046 g, 3.23 × 10⁻⁶) was dispersed in 4 mL of a degassed solution of Milli-Q water and DMSO (85% / 15%, vv.). -4 A solution containing (0.1558 g, 5.70 × 10) of polygalactose azide (5.70 × 10) was injected into a nanoworm dispersion via a degassed syringe. -5 A solution containing 2 mL of degassed Milli-Q water and DMSO (85% / 15%, vv.) was injected into the nanoworm dispersion via a degassed syringe. 4 mL of ascorbic acid (0.4686 g, 2.66 × 10⁻¹⁴) was added. -3 A degassed solution (85% / 15%, vv.) of Milli-Q water and DMSO containing a mol(10) solution was injected into a dispersion of nanoworms, guanidine azide, and polygalactose azide via a degassed syringe. Subsequently, 4 mL of CuSO4 (0.1820 g, 1.14 × 10) was added. -3A degassed solution (85% / 15%, vv.) of Milli-Q water containing a mol(1) solution of DMSO was injected into the reaction mixture via a degassed syringe. The reaction was carried out under an argon atmosphere at 23°C for 19 hours. The reaction was interrupted by exposure to air, and the suspension was dialyzed against 0.01 M EDTA disodium salt (3500 MWCO, 3 × 1 L, alternating every 2 hours), followed by dialyzed against Milli-Q water for 24 hours (3500 MWCO, 6 × 1 L, alternating every 4 hours). The resulting solution was freeze-dried to obtain the product as a light green powder.

[0201]

[0236] Figure 17D shows a schematic diagram illustrating the resulting conjugation of guanidine azide and polygalactose azide to quaternized grafted alkyne PDMAEMA nanoworms. Guanidine azide and polygalactose azide are used in portions 1730 and 1740 of P(NIPAM). 55 -co-DMAEMA 48 It was conjugated to the propargyl group of ). Nanoworm 1700D corresponds to Nanoworm Example 22 (P11).

[0202] Preparation of polygalactose azide

[0237] Step 1. SET-LRP of 6-O-acryloyl-1,2:3,4-di-O-isopropylididene-D-galactopyranose

[0238] Single-electron transfer living radical polymerization was carried out according to the following reaction scheme (XI). TIFF0007872223000026.tif168170

[0203]

[0239] Protected sugar acrylate (0.5g, 1.59 x 10) -3 mol), Me6tren (4.3μL, 0.0037g, 1.59×10 -5 mol), CuBr2 / Me6tren (0.0072g, 1.59×10 -5 A 4 mL vial was filled with mol) and DMSO (1 mL), cooled to 0°C, and purged with argon for 30 minutes to remove oxygen. Cu(0) powder (0.001 g, 1.59 × 10) -5The EBiB (mol) was added to a 15 mL Schlenk tube and purged with argon for 30 minutes. Then, the initiator EBiB (23 μL, 0.031 g, 1.59 × 10) was added. -4 A 1 mL degassed solution containing (mol) DMSO was added to a 4 mL vial via a degassed syringe. The reaction mixture from the 4 mL vial was transferred to a Schlenk tube via a degassed syringe. The Schlenk tube was placed in a temperature-controlled oil bath at 25°C. Samples for SEC were taken at time to monitor the reaction progress. The reaction was interrupted after 4 hours by quenching with liquid nitrogen. Subsequently, 10 mL of acetone was added again to the reaction mixture, and this solution was passed through a basic alumina column. The column was washed several times with acetone. The acetone was evaporated using a rotary evaporator, and the residue was used directly in the azide step.

[0204]

[0240] Step 2. Azidation of protected polygalactose-Br

[0241] The azidation of protected polygalactose-Br was carried out according to the following reaction scheme (XII). TIFF0007872223000027.tif151170

[0205]

[0242] Polymer (0.35g, 1.05 x 10) -4 (mol) was dissolved in 15 mL of DMF. Then NaN 3( 0.1363g, 2.10 × 10 -3 A mol of salt was added to the polymer solution. The reaction was allowed to proceed for 19 hours. DMF was removed by nitrogen flow overnight. The reaction mixture was dissolved again in 12 mL of chloroform, and the undissolved salt was filtered out. Chloroform was removed by nitrogen flow to obtain a white powder as the product.

[0206]

[0243] Step 3. Protect polygalactose-N 3 Deprotection

[0244] Polymer (acetal group) 0.3g, 1.8 x 10 -3 The mol of TFA was dissolved in 4 mL of DCM. Then, 2 mL of TFA (2.978 g, 2.6 × 10) was added. -2(14.4 equivalents, mol) was added. The reaction mixture was stirred for 24 hours. The reaction mixture was then dialyzed against acetone (2 × 500 mL). The acetone was removed by nitrogen flow, and the residue was dried under high vacuum to obtain a brown solid as the product.

[0207] Antibacterial properties of nanoworms

[0245] Live H3N2 influenza virus

[0246] Surfaces coated with nanoworms were tested against live H3N2 influenza enveloped virus according to the following procedure. 10 μL droplets of virus at 1.3 × 10⁵ PFU / mL were added to the nanoworm-coated surface. The virus used was strain A / Switzerland / 9715293 / 2013(H3N2). The surface was incubated at room temperature for 30 minutes, then washed with 190 μL of PBS + trypsin (2 μg / mL). 2-fold serial dilutions of the wash solution and 50 μL of dilution were added to Vero cells in each well. The wells were incubated at 37°C for 1 hour. The virus wash solution was removed, and a stratified medium (M199 medium with 1.5% CMC, 2% FCS, 1:100 PenStrep, 2 μg / mL Trypsin) was added, followed by incubation at 37°C for 72 hours. The stratified and fixed cells were removed by adding 100 μL / well of well-chilled 80% acetone / 20% PBS, and incubated at -20°C for 20 minutes. The acetone was removed, and the plates were air-dried overnight. Immunostain plates were then prepared using 2 μg / mL of hFI6v3 and a 1:2000 dilution of goat anti-human IRDYE800 secondary antibody. Plaque-forming units were counted, and the number of plaque-forming units per ml was calculated. Figure 11 shows an example of antimicrobial activity against H3N2 influenza virus on a nanoworm-coated surface in one embodiment. Each of the nanoworm-coated surfaces (A, B, C, D) showed antimicrobial properties with reduced levels of plaque-forming units compared to the control.

[0208]

[0247] Cloned attenuated AAV-HA virus using a solution assay protocol

[0248] Surfaces coated with nanoworms were tested against cloned attenuated AAV-HA virus according to the following procedure. Solution assays of 0.002, 0.2, and 20 micrograms (μg) of nanoworm polymer were diluted in 250 microliters (μL) of DMEM (FCS-free) medium. 1 μL of AAV-HA solution containing 1.0 × 10⁹ PFU / mL was added and incubated at room temperature for 30 minutes. 250 μL / well was added to HEK293 cells and incubated at 37°C for 1 hour. The virus solution was removed from the cells, DMEM medium was added to the wells, and incubated for a further 1 hour at 37°C. Cells were harvested and genomic DNA was extracted for real-time PCR. Quantification of extracted DNA correlated with pfu / ml. Figure 12 shows an example of antimicrobial activity against AAV-HA virus for nanoworm-coated surfaces in one embodiment. Each of the nanoworm-coated surfaces (A, B, C, D) showed increased antimicrobial properties when the amount of nanoworm polymer was increased to 20 μg.

[0209]

[0249] Cloned attenuated AAV-HA virus by surface assay protocol

[0250] Surfaces coated with nanoworms were tested against cloned attenuated AAV-HA virus according to the following procedure. 10 μL of a 1.0 × 10⁶ PFU / mL surface assay of AAV-HA virus diluted in PBS buffer was added to the nanoworm-coated surface and incubated at room temperature for 30 minutes. The surface was washed with 250 μL of PBS and vortexed five times for 2 seconds each. 250 μL was added to each well containing HEK293 cells and incubated at 37°C for 1 hour. The virus solution was removed from the cells, DMEM medium was added to the wells, and incubated for a further 1 hour at 37°C. Cells were harvested and genomic DNA was extracted for real-time PCR. Quantification of the extracted DNA correlated with pfu / ml. Figure 13 shows an example of antimicrobial activity against AAV-HA virus for a nanoworm-coated surface in one aspect. Each of the nanoworm-coated surfaces (A, B, C, D) showed antimicrobial properties with reduced levels of plaque-forming units compared to the control and compared to the antimicrobial properties of PEI.

[0210]

[0251] Surface sprayed with nanoworms against AAV-HA recombinant virus (capsid)

[0252] Surfaces sprayed with nanoworms were tested against recombinant AAV-HA virus (capsid). A 10 μl solution containing 1,000,000 AAV-HA virus genome copies was buffered to pH 6.5 and applied to various surfaces. Figure 18 shows the decrease in viral activity when applied to various surfaces. Surface 1810 was a control glass slip surface treated with piranha solution and not sprayed with any nanoworm coating. Surface 1820 was an armrest surface not sprayed with any nanoworm coating. Surface 1830 was an armrest surface with the nanoworm coating of Nanoworm Example 17 (P3). Surface 1840 was an armrest surface with the nanoworm coating of Nanoworm Example 18 (P6). Surface 1850 was an armrest surface with the nanoworm coating of Nanoworm Example 20 (P8). Surface 1860 was the surface of an armrest having a nanoworm coating according to Nanoworm Example 21 (P9). Surface 1870 was the surface of an armrest having a nanoworm coating according to Nanoworm Example 19 (P10). Surfaces 1830-1870 were formed by spraying the nanoworm solution five times. Each of the surfaces 1830-1870 with the nanoworm coating showed an increased antibacterial effect compared to surfaces 1810-1820.

[0211]

[0253] Surfaces sprayed with nanoworms were tested against recombinant AAV-HA virus (capsid). A 10 μl solution containing 1,000,000 AAV-HA virus genome copies was buffered to pH 6.5 and applied to various surfaces. Figure 19 shows the decrease in viral activity when applied to various surfaces. Surface 1910 was a control glass slip surface treated with piranha solution and not sprayed with any nanoworm coating. Surface 1920 was an armrest surface not sprayed with any nanoworm coating. Surface 1930 was an armrest surface with the nanoworm coating of Nanoworm Example 17 (P3). Surfaces 1940-1950 were armrest surfaces with the nanoworm coating of Nanoworm Example 20 (P8). Surface 1960 was an armrest surface with the nanoworm coating of Nanoworm Example 21 (P9). Surface 1970 was the surface of an armrest having a nanoworm coating according to Nanoworm Example 19 (P10). Surface 1980 was the surface of an armrest having a nanoworm coating according to Nanoworm Example 22 (P11). Surface 1940 was formed by spraying the nanoworm solution twice. Surfaces 1950-1980 were formed by spraying the nanoworm solution twice. Each of the surfaces 1940-1980 with the nanoworm coating showed an increased antibacterial effect compared to surfaces 1910-1920.

[0212]

[0254] Surfaces sprayed with nanoworms were tested against recombinant AAV-HA virus (capsid). A 10 μl solution containing 1,000,000 AAV-HA virus genome copies was buffered to pH 7.4 and applied to various surfaces. Figure 20 shows the decrease in viral activity when applied to various surfaces. Surface 2010 was a control glass slip surface treated with piranha solution and not sprayed with any nanoworm coating. Surface 2020 was an armrest surface not sprayed with any nanoworm coating. Surface 2030 was an armrest surface with the nanoworm coating of Nanoworm Example 17 (P3). Surface 2040 was an armrest surface with the nanoworm coating of Nanoworm Example 20 (P8). Surface 2050 was an armrest surface with the nanoworm coating of Nanoworm Example 21 (P9). Surface 2060 was the surface of an armrest having a nanoworm coating according to Nanoworm Example 19 (P10). Surface 2070 was the surface of an armrest having a nanoworm coating according to Nanoworm Example 22 (P11). Surfaces 2030-2070 were formed by spraying the nanoworm solution five times. Each of the surfaces 2030-2070 with the nanoworm coating showed an increased antibacterial effect compared to surfaces 2010-2020.

[0213]

[0255] Nanoworm sprayed surface against coronavirus (enveloped virus)

[0256] Surfaces sprayed with nanoworms were tested against SARS-CoV-2 (enveloped virus) capsid. The amount of SARS-CoV-2 sample buffered at pH 6.5 applied to various surfaces is shown. Figure 21 shows the decrease in viral activity after 30 minutes of exposure to different surfaces. Surface 2010 was the surface of an armrest without any nanoworm coating. Surface 2020 was the surface of an armrest with the nanoworm coating of Nanoworm Example 20 (P8). Surface 2030 was the surface of an armrest with the nanoworm coating of Nanoworm Example 22 (P11). Surface 2030 showed a decrease in viral activity below the detection limit. It is thought that Nanoworm Example 22 (P11) showed a higher antiviral effect compared to Nanoworm Example 20 (P8) due to the glycan-mimicking targeting of polysaccharide groups in Nanoworm Example 22, compared to Nanoworm Example 20 (P8), where polysaccharide groups are absent.

[0214]

[0257] The descriptions of the various embodiments of this disclosure are presented for illustrative purposes only and are not intended to be exhaustive or limitful to the embodiments disclosed. A number of modifications and changes will become apparent to those skilled in the art without departing from the scope and spirit of the embodiments described. The terminology used herein has been chosen to best illustrate the principles, practical applications, or technical improvements of embodiments of the art found in the market, or to make the embodiments disclosed herein understandable to others skilled in the art.

Claims

1. It is a nanoworm, Multiple alkene units, R derived from reversible addition-cleavage chain transfer agent １ Multiple first macro CTA polymer units containing a group and Includes, A nanoworm in which the first macro CTA polymer unit contains a quaternized amine.

2. The nanoworm according to claim 1, wherein the plurality of first macro CTA polymer units have a lower critical eutectic temperature (LCST) in water from -20°C to +100°C.

3. The nanoworm according to claim 1 or 2, wherein the plurality of first macro CTA polymer units are configured to respond to temperature and to environmental conditions selected from the group consisting of pH, salinity, and light.

4. The R of the first macro CTA polymer unit １ The nanoworm according to any one of claims 1 to 3, wherein the group is a functional group selected from the group consisting of carboxylic acids, alkynes, pyridines, dopamine, thiolactones, biotin, azides, peptide sequences, sugar sequences, proteases, glycans, polymers, and combinations thereof.

5. The nanoworm according to any one of claims 1 to 4, wherein the first macro CTA polymer unit comprises a functionalized quaternized amine, wherein the first macro CTA polymer unit is functionalized with a functional group selected from alkyl groups, carboxylic acids, alkynes, pyridines, dopamins, thiolactones, biotins, azides, peptide sequences, sugar sequences, proteases, glycans, polymers, and combinations thereof.

6. The nanoworm according to any one of claims 1 to 5, wherein the first macro CTA polymer unit comprises two or more functionalized quaternary amines selected from functional groups consisting of alkyl groups, carboxylic acids, alkynes, pyridines, dopamines, thiolactones, biotins, azides, peptide sequences, sugar sequences, proteases, glycans, polymers, and combinations thereof.

7. The nanoworm according to any one of claims 1 to 6, wherein the first macro CTA polymer unit comprises a functionalized quaternary amine of a first short-chain alkyl quaternary group and a functionalized quaternary amine of a second long-chain alkyl quaternary group, wherein the short-chain alkyl group has 1 to 4 carbon atoms and the long-chain alkyl quaternary group has 5 or more carbon atoms.

8. A plurality of second macro CTA polymer units, wherein the second macro CTA polymer units are derived from a reversible addition-cleavage chain transfer agent R １ It further comprises a plurality of second macro CTA polymer units containing a group, The plurality of first macro CTA polymer units are different from the plurality of second macro CTA polymer units. A nanoworm according to any one of claims 1 to 7.

9. The first macro CTA polymer unit is poly(N-isopropylacrylamide) (PNIPAM), poly(N,N-(dimethylamino)ethyl methacrylate) (F), poly(N-acetoxyethylacrylamide) (PNAEAA), poly(acryloylglycine ethyl ester) (PNAGEE), poly((ethylene glycol) methyl ether methacrylate) (PEGMEMA), poly((propylene glycol) methacrylate) (PPGMA), poly(N,N-dimethylacrylamide) (PDMA), poly(N- A nanoworm according to any one of claims 1 to 8, comprising a polymer selected from the group consisting of decylacrylamide (PDcA), poly(N,N-diethylacrylamide) (PDEA), poly(N-acryloylglycine) (PNAG), poly(N-acryloylglycine methyl ester) (PNAGME), poly(N-acryloylglycine ethyl ester) (PNAGEE), and poly(N-acryloylglycine propyl ester) (PNAGPE), polyacrylamide, polyacrylate, and copolymers thereof.

10. The nanoworm according to any one of claims 1 to 9, further comprising a plurality of grafted polymers grafted onto at least a portion of the plurality of first macro CTA polymer units.

11. The nanoworm according to claim 10, wherein the grafted polymer comprises a functionalized quaternized amine selected from functional groups consisting of alkyl, carboxylic acid, alkyne, pyridine, dopamine, thiolactone, biotin, azide, peptide sequence, sugar sequence, protease, glycanase, and combinations thereof.

12. The nanoworm according to claim 10 or 11, wherein the grafted polymer comprises two or more functionalized quaternary amines selected from functional groups consisting of alkyl, carboxylic acid, alkyne, pyridine, dopamine, thiolactone, biotin, azide, peptide sequence, sugar sequence, protease, glycanase, and combinations thereof.

13. The nanoworm according to any one of claims 10 to 12, wherein the grafted polymer comprises a first functionalized quaternary amine of a short-chain alkyl quaternary group and a second functionalized quaternary amine, wherein the short-chain alkyl group has 1 to 4 carbon atoms and the long-chain alkyl quaternary group has 5 or more carbon atoms.

14. The grafted polymer is A first functionalized quaternized amine group containing a peptide sequence, A second functionalized quaternary amine group containing a sugar sequence and A nanoworm according to any one of claims 10 to 13, including the nanoworm.

15. The grafted polymer is the R of the first macro CTA polymer unit. １ A nanoworm according to any one of claims 10 to 14, grafted onto a substrate.

16. The nanoworm according to any one of claims 10 to 15, wherein the grafted polymer is grafted onto a quaternary amine of the first macro CTA polymer unit.

17. Multiple first grafted polymers are the R of the first macro CTA polymer unit. １ A base is grafted, and a plurality of second grafted polymers are grafted onto the quaternary amine of the first macro CTA polymer unit. A nanoworm according to any one of claims 10 to 16.

18. The nanoworm according to any one of claims 10 to 17, wherein the grafted polymer is formed by a polymerization method selected from the group consisting of addition polymerization, chain polymerization, radical polymerization, metal-catalyzed polymerization, nitroxide polymerization, exchange chain transfer polymerization, RAFT, SET-LRP, condensation polymerization, and combinations thereof.

19. A nanoworm according to any one of claims 10 to 18, comprising 20 to 400 alkene units, 1 to 200 first macro CTA polymer units, and 1 to 10,000 grafted polymers.

20. A nanoworm according to any one of claims 1 to 19, comprising a peptide sequence capable of inhibiting virus attachment and virus-cell membrane fusion.

21. A nanoworm according to any one of claims 1 to 20, comprising a peptide sequence capable of disrupting the viral envelope.

22. A nanoworm according to any one of claims 1 to 21, comprising a peptide sequence capable of inhibiting viral replication.

23. A nanoworm according to any one of claims 1 to 22, comprising a quaternized alkylquaternary ammonium cation capable of killing bacteria.

24. A composition comprising one or a combination of the nanoworms described in any one of claims 1 to 23.

25. A coating comprising one or a combination of the nanoworms described in any one of claims 1 to 23.

26. The coating according to claim 25, which can be washed and reused to replenish the antibacterial properties of the nanoworms.

27. A transporter comprising the coating according to claim 25 or 26.

28. An object comprising the coating according to claim 25 or 26.

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