Antipathogenic nanostructures
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- THE BOEING CO
- Filing Date
- 2024-05-15
- Publication Date
- 2026-05-29
Smart Images
Figure 2026517422000001_ABST
Abstract
Description
[Technical Field]
[0001] Related applications
[0001] This application claims the benefit and priority of U.S. Provisional Patent Application No. 63 / 467746, filed on 19 May 2023 (the entire contents of which are incorporated herein by reference).
[0002]
[0002] The present disclosure provides antipathogenic nanostructures such as coatings of antipathogenic nanostructures that cover the surface of vehicles, buildings, clothing, filters or any other suitable objects. [Background technology]
[0003]
[0003] Pandemics have long-term and devastating effects on society. Reducing the potential for transmission of pathogens such as viruses and microorganisms can reduce the transmission of pathogens on highly contacted surfaces and in enclosed environments such as vehicles, offices, transportation facilities, and homes.
[0004]
[0004] One of the goals of airlines is the cleanliness of aircraft cabins and space transport, and the habitat-related industries are interested in reducing the possibility of transmission of pathogens such as viruses and microorganisms. In fact, certain pathogens, such as mRNA pathogens like SARS-CoV-2 and its variants, are sense RNA viruses. Sense RNA viruses can translate proteins even after the host cell has been lysed. The characteristics of such mRNA pathogens make it difficult to prevent the transmission of pathogens because standard cell lysis techniques are often ineffective against these sense RNA viruses. Furthermore, these lysis techniques often require the use of undesirable chemicals to synthesize the molecules used to lyse pathogenic cells.
[0005]
[0005] Currently, disease transmission reduction technologies focus on improving the mechanical characteristics of air filtration systems. Unfortunately, this cannot reduce or stop the transmission of pathogens through surfaces. Other disease reduction technologies focus on lysing host cells carrying pathogens. However, these technologies may not be able to inactivate the RNA and RNA translation of sense RNA viruses. Therefore, sense RNA viruses can remain on the surface.
[0006]
[0006] Therefore, there is a need for an improved self-disinfecting surface coating agent.
Summary of the Invention
[0007]
[0007] The present disclosure provides a nanostructure comprising a compound or a salt thereof. The compound comprises a plurality of N-alkylacrylamide units. The compound has the following formula: TIFF2026517422000002.tif19170(where R 1 is C1-C 20 alkyl). The compound has the following formula: TIFF2026517422000003.tif28170(where R 2 , R 3 and R 4 are each independently hydrogen or C1-C 20 alkyl). The compound has the following formula: TIFF2026517422000004.tif20170(where R 5 is C1-C 10 aryl). The compound has a plurality of parts represented by the following formula: TIFF2026517422000005.tif61170(where Q is O or N, R ** is C1-C 20 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc., and R 6 , R 7 and R 8R is independently a C1-C6 alkyl group or hydrogen atom. 9 If present, C1-C 16 Chromophores such as alkyl, C1-C6 alkylene (alkylyne), azole, diguanidine, polysaccharide, coumarin, and one or more combinations thereof, where q is an integer of 0 or 1, and in at least one of the multiple examples R 9 (There exists an oligomer of guanidine (e.g., diguanidine)) It includes multiple parts represented by .
[0008]
[0008] The disclosure also provides a method for depositing nanostructures on a surface. In some embodiments, the nanostructure includes a compound or a salt thereof. The compound includes a plurality of N-alkylacrylamide units. The compound has the following formula: TIFF2026517422000006.tif19170 (in the formula, R 1 is C1-C 20 The compound contains a portion represented by the alkyl group. The compound is given by the following formula: TIFF2026517422000007.tif28170 (in the formula, R 2 , R 3 and R 4 Each is independently hydrogen or C1-C 20 The compound contains a portion represented by the alkyl group. The compound is given by the following formula: TIFF2026517422000008.tif23170 (in the formula, R 5 is C1-C 10 The compound contains multiple parts represented by aryl compounds. The compound is given by the following formula: TIFF2026517422000009.tif61170(In the formula, Q is O or N, R ** is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. 6 , R 7 and R 8 R is a C1-C6 alkyl group or hydrogen, 9 If present, C1-C16 Alkyl, C1-C6 alkylene, azole, guanidine, guanidine oligomer (e.g., diguanidine), polysaccharide, chromophore, and one or more combinations thereof, where q is an integer of 0 or 1, and in at least one of the multiple examples R 9 (There exists an oligomer of guanidine (e.g., diguanidine)) It includes multiple parts represented by .
[0009]
[0009] To enable a more detailed understanding of the features of the Disclosure described above, a more specific description of the Disclosure, which is briefly summarized above, can be obtained by referring to the embodiments. Some embodiments are shown in the accompanying drawings. However, it should be noted that the accompanying drawings show only typical embodiments of the Disclosure and are therefore not limiting to its scope, as the Disclosure may recognize other equally effective embodiments. [Brief explanation of the drawing]
[0010] [Figure 1A]
[0010] This is a schematic diagram showing a nanoworm according to a particular aspect of the present disclosure. [Figure 1B]
[0011] A diagram of a synthesis scheme for forming nanostructures according to a particular aspect of this disclosure is shown. [Figure 2]
[0012] The 1H NMR (CDCl3) spectrum of a crude mixture of 1,1'-(azandiylbis(propane-3,1-diyl))diguanidine hydrobromide according to a particular aspect of this disclosure is shown. [Figure 3]
[0013] The 1H NMR (CDCl3) spectrum of a crude mixture of 1,1'-(azandiylbis(propane-3,1-diyl))diguanidine hydrobromide according to a particular aspect of this disclosure is shown. [Figure 4]
[0014] The ¹H NMR (CDCl3) spectrum of a crude mixture of 2-chloro-N,N-bis(3-guanidinopropyl)acetamide hydrobromide according to a particular aspect of this disclosure is shown. [Figure 5]
[0015] The ¹H NMR (CDCl3) spectrum of a crude mixture of 2-chloro-N,N-bis(3-guanidinopropyl)acetamide hydrobromide according to a particular aspect of this disclosure is shown. [Figure 6]
[0016] The 1H NMR (CDCl3) spectrum of a crude mixture of macro CTA-A immediately after polymerization (i.e., before dialysis) according to a particular aspect of this disclosure is shown. [Figure 7]
[0017] The molecular weight distribution of macro CTA-A, determined by SEC (RI detection, using DMAc as the eluent) according to a particular aspect of this disclosure, is shown. [Figure 8]
[0018] The 1H NMR (CDCl3) spectra of macro-CTA-A after purification by dialysis and freeze-drying in water, according to a particular aspect of this disclosure, are shown. [Figure 9]
[0019] The 1H NMR (CDCl3) spectrum of the crude mixture of macro CTA-B immediately after polymerization (i.e., before dialysis) is shown. The DMAEMA conversion rate is 100%. According to certain embodiments of this disclosure, the NIPAM conversion rate is 90%. [Figure 10]
[0020] The molecular weight distribution of macro CTA-B determined by SEC (RI detection, using DMAc as the eluent) according to a particular aspect of this disclosure is shown. [Figure 11]
[0021] The 1H NMR (in CDCl3) spectrum of macro CTA-B after purification by dialysis and freeze-drying in water, according to a particular aspect of this disclosure, is shown. According to a particular aspect of this disclosure, n(NIPAM)=45; n(DMAEMA)=30. [Figure 12]
[0022] The following describes a procedure for synthesizing nanoworms according to a particular aspect of this disclosure. [Figure 13]
[0023] The ¹H NMR (CDCl3) spectrum of a crude mixture obtained by emulsion polymerization according to a particular aspect of this disclosure is shown. [Figure 14]
[0024] The SEC trace of the purified polymer after dialysis and lyophilization is shown (RI detector, DMAc as eluent). According to certain aspects of this disclosure, Mn,TD = 19389, DRI 1.36. [Figure 15]
[0025] The 1H NMR (DMSO-d6) spectra of the purified emulsion latex after dialysis and lyophilization, according to a particular aspect of this disclosure, are shown. [Figure 16A-C]
[0026] TEM images of the nanostructure according to a specific aspect of this disclosure are shown. Figure 16A shows nanoworms (spherical particles) without toluene addition. Figure 16B shows the nanostructure (nanoworms) after toluene addition. Figure 16C shows the removal of toluene from the nanoworms by rotary evaporation (the nanoworm structure is maintained). [Figure 17]
[0027] The following describes a procedure for synthesizing nanoworms according to a particular aspect of this disclosure. [Figure 18]
[0028] The 1H NMR (CDCl3) spectrum of the polymer latex collected after emulsion polymerization at 70°C for 5 hours, exposure to air at 70°C for 4 hours, and cooling from 70°C to 25°C for 24 hours, according to a particular aspect of the present disclosure, is shown. [Figure 19]
[0029] The images show TEM images of nanostructured spherical particles produced after (i) emulsion polymerization at 70°C for 5 hours, (ii) exposure of the polymer latex to air at 70°C for 4 hours, and (iii) cooling of the polymer latex from 70°C to 25°C for 24 hours, according to a particular aspect of the present disclosure. [Figure 20A-B]
[0030] Figure 20A shows TEM images of nanoworms (NW) heated at 50°C for 1 hour according to a particular aspect of this disclosure. Figure 20A shows 8.2 wt% nanoworms in water. Figure 20B shows 4.1 wt%. [Figure 21]
[0031] This disclosure describes a synthetic process for producing nanoworms using PGal, poly(D-galactose 6-O-acrylate), 7HC-N3, and 3-azido-7-hydroxycoumarin, according to a particular aspect of this disclosure. [Figure 22]
[0032] This disclosure describes a specific aspect of the synthesis process for quaternization of nanoworms using diguanidine chloroacetamide (digua-Cl) at 60°C. [Figure 23]
[0033] The 1H NMR (DMSO-d6) spectra of the crude reaction mixture before and after the reaction are shown according to a particular aspect of this disclosure. [Figure 24]
[0034] The 1H NMR (DMSO-d6) spectra of the purified polymer before and after quaternization using digua-Cl (post-dialysis) according to a particular aspect of this disclosure are shown. [Figure 25]
[0035] This disclosure describes a synthetic process for quaternization of nanoworms at 50°C with diguanidine chloroacetamide prior to functionalization using propargyl bromide, 1-iodooctane, polysugar azide, and coumarin azide, according to a particular aspect of this disclosure. [Figure 26]
[0036] The 1H NMR (DMSO-d6) spectrum of quaternization of nanoworms at 50°C with di-gua-Cl according to a particular aspect of this disclosure is shown. [Figure 27A-B]
[0037] Figure 27A shows TEM images of nanoworms after reaction with digua-Cl at 50°C according to a particular aspect of this disclosure. Figure 27A shows the reaction after 14 hours. Figure 27B shows the reaction after 36 hours. [Figure 28]
[0038] This disclosure describes a specific aspect of the synthesis process for quaternization of nanoworms using propargyl bromide. [Figure 29]
[0039] The ¹H NMR (DMSO-d6) spectrum for the quaternization of nanoworms at room temperature with propargyl bromide according to a particular aspect of this disclosure is shown. The enlarged portion focuses on the CH2 peak of propargyl bromide. Reactions A and B were carried out simultaneously. The toluene detected by NMR was from the propargyl bromide solution. [Figure 30]
[0040] The 1H NMR (DMSO-d6) spectrum of the purified sample after quaternization with propargyl bromide, according to a particular aspect of this disclosure, is shown. [Figure 31]
[0041] This disclosure describes a specific aspect of the synthesis process for quaternization of nanoworms using 1-iodooctane. [Figure 32]
[0042] The 1H NMR (in DMSO-d6) spectra of the purified sample after further quaternization with 1-iodooctane, according to a particular aspect of this disclosure, are shown in comparison with those of the purified spherical nanoparticles. [Figure 33]
[0043] This disclosure describes a specific process for synthesizing copper-catalyzed azido-alkyne cycloaddition (CuAAC) involving the bonding of polysaccharide azides and coumarin azides. [Figure 34]
[0044] The 1H NMR spectra (in DMSO-d6) of the crude reaction mixture before and after CuAAC, according to a particular aspect of this disclosure, are shown. [Figure 35]
[0045] This is a schematic diagram of the click reaction to nanoworms according to a particular aspect of the present disclosure. [Figure 36A-B]
[0046] Post-dialysis TEM images are shown for two replicate reactions according to a particular aspect of this disclosure. Figure 36A shows the post-dialysis TEM image of reaction A. Figure 36B shows the post-dialysis TEM image of reaction B. [Figure 37]
[0047] In accordance with a particular aspect of this disclosure, a comparison of 1H NMR (in DMSO-d6) spectra between samples (one of parallel reactions) before and after CuAAC for tracking polysaccharides is shown. [Figure 38]
[0048] In accordance with a particular aspect of this disclosure, a comparison of 1H NMR (in DMSO-d6) spectra between samples before and after CuAAC (the other side of a parallel reaction) for tracking polysaccharides is shown. [Figure 39A]
[0049] Figures 39A to 39C show an analysis of a nanostructure according to a particular aspect of the present disclosure. Figure 39A shows a compound of the nanostructure. [Figure 39B] The 1H NMR spectrum of the purified sample from reaction A is shown. [Figure 39C] The 1H NMR spectrum after adding D2O to the DMSO-d6 NMR mixture is shown. [Figure 40]
[0050] The 1H NMR spectra of the purified sample of reaction B, one of the reactions obtained by replication experiments according to a particular aspect of this disclosure, and the sample after adding D2O to the DMSO-d6 NMR mixture are shown. [Figure 41]
[0051] This disclosure describes a synthetic process for quaternization of nanoworms at 50°C with diguanidine chloroacetamide, following functionalization using propargyl bromide, 1-iodooctane, polysaccharide azide, and coumarin azide, according to a particular aspect of this disclosure. [Figure 42A-B]
[0052] TEM images of nanostructures according to specific embodiments of this disclosure are shown. Figure 42A shows a TEM image after quaternization with digua-Cl. Figure 42B shows TEM images of two parallel reactions. [Figure 43A]
[0053] Figures 43A-43C show the analysis of nanostructures according to a particular aspect of this disclosure. Figure 43A shows the 1H NMR spectrum (in DMSO-d6) of purification reaction A after quaternization using digua-Cl at 50°C. [Figure 43B] The 1H NMR spectrum (in DMSO-d6) of purification reaction B after quaternization using digua-Cl at 50°C is shown. [Figure 43C] The 1H NMR spectrum (in DMSO-d6) of purified nanoworms after quaternization using digua-Cl at 50°C is shown. [Figure 44]
[0054] This disclosure describes a specific aspect of a synthetic process for generating nanostructures via quaternization of nanoworms at high temperatures using proposed polysaccharide pseudohalides, digua-Cl, and iodooctane components. [Figure 45A-E]
[0055] Figures 43A–45F illustrate synthesis processes for generating nanostructures via quaternization of nanoworms according to specific embodiments of this disclosure. Figure 45A shows a first synthesis process. Figure 45B shows a second synthesis process. Figure 45C shows a third synthesis process. Figure 45D shows a fourth synthesis process. Figure 46E shows a fifth synthesis process. [Figure 45F] This shows a typical synthesis process. [Figure 46]
[0056] This disclosure describes a synthetic process for generating nanostructures via quaternization of nanoworms, according to a particular aspect of this disclosure. [Figure 47A-C]
[0057] Figure 47A shows a TEM image of a nanostructure according to a specific aspect of the present disclosure. Figure 47A shows a TEM image after quaternization without the addition of a plasticizer. Figure 47B shows a TEM image of a parallel reaction after quaternization without the addition of a plasticizer. Figure 47C shows a TEM image of a parallel reaction after quaternization without the addition of a plasticizer. [Figure 48]
[0058] This disclosure describes a synthesis process for CuAAAC "click" nanoworms using polysaccharides and chromophores according to a particular aspect of this disclosure. [Figure 49A-C]
[0059] TEM images of nanostructures according to specific embodiments of this disclosure are shown. Figure 49A is a TEM image after CuAAC using a polysaccharide and a chromophore. Figure 49B is a TEM image of a parallel reaction after CuAAC using a polysaccharide and a chromophore. Figure 49C is a TEM image of a parallel reaction after CuAAC using a polysaccharide and a chromophore. [Figure 50]
[0060] This is a schematic diagram of the conversion from spherical particles to nanoworms according to a particular aspect of the present disclosure. [Figure 51A-B]
[0061] TEM images of nanostructures according to specific embodiments of this disclosure are shown. Figure 51A shows nanoworms obtained from a first mixture. Figure 51B shows nanoworms obtained from a second mixture. [Figure 52]
[0062] This disclosure describes a large-scale synthesis procedure for nanoworms according to a specific aspect of this disclosure. [Figure 53]
[0063] This disclosure shows an SEC trace of a large nanoworm in a specific aspect. [Figure 54]
[0064] This disclosure describes a large-scale synthetic procedure for quaternization of nanoworms according to a specific aspect of this disclosure. [Figure 55A-C]
[0065] Figure 55A shows a TEM image of a nanostructure according to a specific aspect of this disclosure. Figure 55A shows a TEM image after quaternization without the addition of a plasticizer. Figure 55B shows a TEM image of a parallel reaction after quaternization without the addition of a plasticizer. Figure 55C shows a TEM image of a parallel reaction after quaternization without the addition of a plasticizer. [Figure 56A-D]
[0066] Figure 56A shows a sample on a tray of a nanostructure according to a specific aspect of this disclosure. Figure 55B shows a sample on a tray of a control sample under ambient light. Figure 56C shows a sample on a tray of a sample coated with nanostructure A under ambient light. Figure 55D shows a sample on a tray of a sample coated with nanostructure A under UV light. [Figure 57]
[0067] In accordance with a particular aspect of this disclosure, the TCID50 spectra of the titer of an omicron mutant of SARS-CoV-2, comparing an uncoated sample with a control sample, are shown. [Figure 58]
[0068] In accordance with certain aspects of this disclosure, qRT-PCR of omicron mutants of SARS-CoV-2 is shown, comparing uncoated samples, comparative samples, and the nanostructures of this disclosure. [Figure 59]
[0069] In accordance with a particular aspect of this disclosure, qRT-PCR of a delta mutant of SARS-CoV-2 is shown comparing an uncoated sample with the nanostructure of this disclosure. [Figure 60]
[0070] In accordance with a particular aspect of this disclosure, qRT-PCR of an alpha mutant of SARS-CoV-2 is shown comparing an uncoated sample with the nanostructure of this disclosure. [Figure 61]
[0071] In accordance with a particular aspect of this disclosure, the TCID50 spectra of influenza infectivity titers are shown, comparing an uncoated sample with a comparison sample. [Figure 62]
[0072] In accordance with a particular aspect of this disclosure, qRT-PCR of the PR8 genome is shown comparing an uncoated sample, a comparison sample, and the nanostructure of this disclosure. [Modes for carrying out the invention]
[0011]
[0073] The global surge in outbreaks of the disease is due to an increase in coronavirus variants and other sense RNA or DNA viruses. For example, mutations in the spike protein on the surface of the virion membrane not only allow for greater transmission but also raise concerns about vaccine effectiveness. To prevent SARS-CoV-2, SARS-CoV-2 variants, and other sense RNA viruses from spreading from person to person through airborne or surface transmission, the viruses must be inactivated.
[0012]
[0074] This disclosure provides nanostructures such as coatings of antipathogenic nanoworms on the surface of vehicles, buildings, wearable articles, filters, or any other suitable objects (e.g., solids and / or those having a porous or non-porous composition such as woven or nonwoven fabrics). Here, the antipathogenic nanoworms are nanoworms suitable for reducing one or more pathogens on the surface to be coated. The coating has antipathogenic properties that are effective in reducing or eliminating pathogens and / or reducing the transmission of pathogens.
[0013]
[0075] The coating can be applied by any aqueous method, such as aqueous spray-on nanocoating. This spray-on nanocoating can inactivate proteins or virion particles and degrade viral DNA or RNA. Although not bound by theory, it is thought that the nanostructure of this coating binds to the viral membrane, and subsequent large conformational changes of the nanoworms cause the nanoworms to rupture the viral membrane. Subsequently, the nanostructure of this coating binds to the genetic material of the virus, degrading it and inactivating viruses such as SARS-CoV-2 (VIC01), the evolved B.1.1.7 (alpha) mutant, influenza A, or viruses containing surrogate capsid pseudoviruses that express hemagglutinin, the adherent glycoprotein of influenza A virus. Polygalactose functional groups on the nanostructure are known to catalyze the degradation of the viral RNA genome by targeting the conserved S2 subunit of the SARS-CoV-2 virion surface spike glycoprotein for stronger binding, and further by attaching guanidine groups.
[0014]
[0076] In several examples, the nanostructures of this disclosure are coated onto the surface of personal protective equipment such as masks, face shields, rebreathers, filter cartridges, or combinations thereof. By coating surgical masks with the nanostructures, enveloped VIC01 and B.1.1.7 can be completely inactivated, providing a powerful control measure against SARS-CoV-2 and its variants. Inactivation of enveloped influenza A virus and AAV-HA capsid pseudovirus has also been observed, demonstrating that broad viral inactivation is possible when using the nanoworms of this disclosure. The technology described herein is a coating with a proposed nanomechanical mechanism for inactivating both enveloped and capsid viruses. This functionalized nanostructure can be modified to target other known and unknown viruses and is suitable for large-scale manufacturing processes.
[0015]
[0077] In certain embodiments, a surface coated with nanostructures can be hydrophilic. For example, a surface coated with nanostructures can be hydrophilic (water-soluble), allowing droplets such as mucosal droplets, blood, urine, sweat, other bodily fluids, and other non-bodily fluids to wet the surface. In certain embodiments, pathogens suspended on or within a droplet can be captured and inactivated by the nanostructure-coated surface. The coatings described herein contain polymers and may have a transparent appearance when applied to a surface. In some embodiments, the coating is useful for inactivating one or more variants of SARS-CoV-2, e.g., all variants. While not bound by theory, it is thought that the coating targets the highly glycosylated spike protein on the virion surface and / or hemagglutinin, the adherent glycoprotein of influenza A virus, disrupting the viral membrane through a process of conformational change of the nanoworm, causing mechanical rupture of the viral membrane.
[0016]
[0078] In certain embodiments, the nanostructures of the present disclosure are prepared using diguanidine reagents (e.g., diguanidine chloroacetamide (digua-Cl), diguanidine acetate, diguanidine phosphate, diguanidine platinum, etc.) that simplify the synthesis process compared to conventional nanostructures. Diguanidine reagents are preferred because they are more stable than other guanidine reagents (e.g., guanidine azide). For example, but not limited to, diguanidine chloroacetamide exhibits higher stability than guanidine azide. Furthermore, or alternatively, diguanidine reagents such as diguanidine chloroacetamide can be mass-produced to ensure commercial applicability.
[0017]
[0079] In certain embodiments, personal protective equipment (e.g., face masks, face shields, rebreathers, filter cartridges, or combinations thereof) coated with the antiviral coating of the Disclosure and treatment of high-contact surfaces with the antiviral coating of the Disclosure can provide long-term disinfection (e.g., days, weeks, months, etc.) of contaminated surfaces, thereby reducing or eliminating the spread of SARS-CoV-2 and / or its variants.
[0018] definition
[0080] The term "DNA" refers to a polymer composed of two polynucleotide chains that wrap around each other to form a double helix. Also known as deoxyribonucleic acid, DNA contains one or more of the following: adenine, cytosine, guanidine, and thymine. DNA may also contain modified bases, such as 5-methylcytosine, N6-carbamoylmethyladenine, N6-methyladenine (methadenine), 7-deazaguanine, 7-methylguanine, N4-methylcytosine, 5-carboxylcytosine, 5-formylcytosine, 5-glycosylhydroxymethylcytosine, 5-hydroxycytosine, 5-methylcytosine, alpha-glutamythymidine, alpha-ptresinylthymine, base j, uracil, 5-dihydroxypentauracil, 5-hydroxymethyldeoxyuracil, deoxyalkaeosine, 2,6-diaminopurine, or combinations thereof. DNS viruses include double-stranded DNA viruses and single-stranded DNA viruses.
[0019]
[0081] As used herein, "nanostructure" refers to a three-dimensional structure provided by two or more functional groups (which may have the same or different chemical structures). The nanostructure ranges from approximately 1 nm to approximately 100 μm. For example, approximately 1 nm to approximately 5 μm, approximately 500 nm to approximately 1 μm, or approximately 10 nm to approximately 250 nm. It is any suitable nanostructure having length or width.
[0020]
[0082] A "nanoworm" is an example of a nanostructure having a high aspect ratio (length ÷ width), where a high aspect ratio means the length of the nanostructure is more than approximately 1000 times its width. A "nanorod" is an example of a nanostructure having a lower aspect ratio (length ÷ width) compared to a nanoworm, where a low aspect ratio means the length of the nanostructure is approximately 10 to approximately 1000 times its width. As used herein, the term "pathogen" refers to viruses, bacteria, fungi, and / or other microorganisms or pathogens. The coating agents described herein can reduce or eliminate the presence and / or transmission of a wide range of pathogens, such as SARS-CoV-2 and its variants, e.g., alpha, beta, delta, omicron, or combinations thereof.
[0021]
[0083] The term “pharmaceutically acceptable” means that it is suitable for use in a pharmaceutical formulation, is generally considered safe for such use, is formally approved by a national or state regulatory authority for such use, or is listed in the United States Pharmacopeia or other generally recognized pharmacopoeias for use in animals, more specifically in humans.
[0022]
[0084] The term "pharmaceutically acceptable salt" refers to a salt that can enhance the desired pharmacological activity. Examples of pharmaceutically acceptable salts include acid addition salts, metal salts, and amine salts produced by reactions with inorganic or organic acids. Examples of acid addition salts with inorganic acids include salts with hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Examples of acid addition salts with organic acids include salts with acetic acid, propionic acid, hexanoic acid, heptanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, o-(4-hydroxybenzoyl)-benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, and p-chloro Examples of salts with benzenesulfonic acid, 2-naphthalenesulfonic acid, p-toluenesulfonic acid, camphorsulfonic acid, 4-methyl-bicyclo[2.2.2]octo-2-ener-carboxylic acid, glucoheptonic acid, 4,4'-methylenebis(3-hydroxy-2-naphthoic acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid. Examples of metal salts include salts with sodium ions, potassium ions, calcium ions, magnesium ions, aluminum ions, iron ions, and zinc ions. Examples of amine salts include salts with ammonia and salts with organic nitrogen bases strong enough to produce salts with carboxylic acids.
[0023]
[0085] The term "RNA" refers to ribonucleic acid present in viruses or cells. RNA may contain one or more of adenine, cytosine, guanine, and / or uracil. RNA may contain phosphate groups attached to the 3' position of one ribose and the 5' position of the next ribose. RNA may also be sense RNA. The term "sense RNA virus" refers to a type of virus that contains either positive sense single-stranded RNA or negative sense single-stranded RNA. Sense RNA viruses can function as mRNA and be directly translated into proteins within the host.
[0024]
[0086] The term "therapeutic dose" refers to the amount of a compound administered to a subject to treat a particular condition that is sufficient to achieve treatment for that condition. The therapeutic dose can vary depending on the compound, the condition and its severity, age, and the body weight of the subject being treated.
[0025]
[0087] The term "virus" typically refers to a supermicroscopic infectious agent comprising a non-living complex molecule that includes a protein coat surrounding a core of RNA or DNA genetic material, lacks a semipermeable membrane, has the ability to grow and reproduce within living cells, and can cause disease in humans, animals, or plants. In at least one embodiment, the virus may be a sense RNA virus, such as a positive sense RNA virus or a negative sense virus. For example, the virus may be SARS-CoV-2 or a variant thereof.
[0026]
[0088] The compounds of this disclosure include tautomers, geometric isomers, or stereoisomers of the compounds. Furthermore, esterified, oxime, onium, hydrated, solvated, and N-oxide forms of the compounds are also included in this disclosure. This disclosure considers all such compounds, including cis and trans geometric isomers (Z- and E-geometric isomers), R- and S-enantiomers, diastereomers, d-isomers, l-isomers, atropisomers, epimers, conformational isomers, rotational isomers, mixtures of isomers, and racemates thereof.
[0027]
[0089] As used herein, the term “SARS-CoV-2 mutant” refers to a virus that has mutated from SARS-CoV-2. Mutations can include approximately 1 to 75 mutations across the entire viral genome, e.g., approximately 25 to 50 mutations. One or more mutations may include mutations in the viral spike protein, e.g., approximately 1 to 40 mutations in the spike protein, e.g., approximately 32 mutations. While not theoretically bound, certain known mutants are thought to have enhanced binding to the ACE2 receptor, primarily through receptor-binding domains on the spike protein found in human throat and lung cells. Upon cell binding, mutations near the S1 / S2 region of the SARS-CoV-2 spike glycoprotein further facilitate cleavage by cell surface serine proteases (e.g., TMPRSS2), exposing the hydrophobic region of the spike to fuse and release viral RNA within the cell, or facilitating intercellular fusion in large multinucleated cells. Different mutants may exhibit different responses to vaccination, different transmission rates, and different symptoms during infection. Antigenic shifts can result from numerous mutations in specific variants, such as omicron spikes, and from widespread replication in immunocompromised hosts or from cross-transmission between humans and rodents. In some embodiments, infected hosts may release SARS-CoV-2 into the environment through sneezing, coughing, or skin contact, resulting in the transmission of the virus to surrounding surfaces. Infectious SARS-CoV-2 has been shown to persist on many different surfaces in laboratory-based studies.
[0028] compound
[0090] The compounds described herein comprise a plurality of N-alkylacrylamide units. In at least one embodiment, which can be combined with other embodiments described herein, the N-alkylacrylamide is of the following formula: TIFF2026517422000010.tif34170 (in the formula, R 10 and R 11 Each of them independently consists of hydrogen or C1-C 20 It is alkyl, R 10 or R 11 At least one of them is C1-C20 Represented by an alkyl group (e.g., methyl, ethyl, n-propyl, or isopropyl). In at least one embodiment which can be combined with other embodiments described herein, R 10 or R 11 At least one of them is isopropyl.
[0029]
[0091] In at least one embodiment that can be combined with other embodiments described herein, the compound is of the formula TIFF2026517422000011.tif19170 (in the formula, R 1 is C1-C 20 It includes a portion represented by (which is alkyl). In some embodiments that can be combined with other embodiments described herein, R 1 These include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl. In some embodiments which can be combined with other embodiments described herein, R 1 It is butyl.
[0030]
[0092] In at least one embodiment that can be combined with other embodiments described herein, the compound is of the formula TIFF2026517422000012.tif32170 (in the formula, R 2 , R 3 and R 4 Each is independently hydrogen or C1-C 20 It includes a portion represented as an alkyl group. In some embodiments, R 2 , R 3 and R 4 Each is independently hydrogen. In some embodiments which can be combined with other embodiments described herein, R 2 , R 3 , and R 4 Each is independently C1-C 20Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 and R 3 is hydrogen, R 4 is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 and R 4 is hydrogen, R 3 is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 3 and R 4 is hydrogen, R 2 is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 is hydrogen, R 4 and R 3 C1-C is independent 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 3 is hydrogen, R 4 and R 2 C1-C is independent 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 and R 4 is methyl, and R 3is hydrogen. In some embodiments that can be combined with other embodiments described herein, R 4 is hydrogen, and R 3 and R 2 are independently C1-C 20 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments that can be combined with other embodiments described herein, R 2 and R 3 are methyl, and R 4 is hydrogen.
[0031]
[0093] In at least one embodiment that can be combined with other embodiments described herein, the compound has the formula TIFF2026517422000013.tif21170 (where R 5 is C4-C 10 allyl). In some embodiments that can be combined with other embodiments described herein, R 5 is C6 aryl. In some embodiments that can be combined with other embodiments described herein, R 5 is phenyl. In some embodiments that can be combined with other embodiments described herein, phenyl can be independently substituted with 1, 2, 3, 4, or 5 moieties such as C1-C 10 alkyl moieties. In at least one embodiment that can be combined with other embodiments described herein, the compound has the following formula: TIFF2026517422000014.tif61170 (where Q is O or N, R ** is C1-C 20 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc., and R 6 , R 7 and R 8 are independently C1-C6 alkyl or hydrogen). In some embodiments that can be combined with other embodiments described herein, R6 , R 7 , and R 8 are each independently C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments, R 6 , R 7 and R 8 are each methyl. In some embodiments that can be combined with other embodiments described herein, R 6 and R 7 are hydrogen, and R 8 is C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments that can be combined with other embodiments described herein, R 6 and R 8 are hydrogen, and R 7 is C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments that can be combined with other embodiments described herein, R 7 and R 8 are hydrogen, and R 6 is C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments that can be combined with other embodiments described herein, R 6 is hydrogen, and R 7 and R 8 are each independently C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments that can be combined with other embodiments described herein, R 7 is hydrogen, and R 6 and R 8 are each independently C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments that can be combined with other embodiments described herein, R 8 is hydrogen, and R 6 and R 7 are each independently C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc.
[0032]
[0094] q is 0 or 1. In some embodiments which can be combined with other embodiments described herein, R 9 At least one of (multiple) is C1-C 16 Alkyls include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl. Although not bound by theory, R 9 If the alkyl group is C5 or higher, the alkyl moiety can more easily penetrate the hydrophobic portion of the cell membrane (such as that of a viral cell), potentially accelerating the lysis of viral cells. Furthermore, the cationic nitrogen moiety can provide Coulomb interactions between the compound (such as the quaternary ammonium moiety) and the cell membrane surface (such as the phosphate portion of the phospholipid bilayer), further promoting the lysis of viral cells. In addition, the quaternary ammonium salt can confer sufficient hydrophilicity, thereby preventing the alkyl group bonded to the quaternary ammonium salt from being substantially embedded within the three-dimensional structure (for example, if the composition has a three-dimensional structure of a nanoworm or nanorod).
[0033]
[0095] In some embodiments that can be combined with other embodiments described herein, R 9 These are C1-C6 alkylenes, such as C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, etc. In some embodiments, R 9 R is an azole. In some embodiments which can be combined with other embodiments described herein, R 9 R is an oligomer of guanidine, for example, diguanidine. In some embodiments which can be combined with other embodiments described herein, 9 The following structure: This is phenyl represented by TIFF2026517422000015.tif56170.
[0034]
[0096] In some embodiments that can be combined with other embodiments described herein, R 9 The following structure: TIFF2026517422000016.tif62170(In the formula, x is an integer from 1 to 20, for example an integer of 10, and R* is hydrogen, -OH, or It is a polygalactose that has TIFF2026517422000017.tif20170.
[0035]
[0097] In some embodiments that can be combined with other embodiments described herein, R 9 R is a coumarin such as 7-hydroxycoumarin. In some embodiments which can be combined with other embodiments described herein, 9 R is a combination of azole and polygalactose (e.g., polygalactose-substituted azole). In some embodiments which can be combined with other embodiments described herein, 9 This is a combination of azole and coumarin (e.g., 3-azido-7-hydroxycoumarin), and is given by the following formula: It has TIFF2026517422000018.tif23170. For example, though not limited to, q is 1 and R 9 It is diguanidine.
[0036]
[0098] In some embodiments that can be combined with other embodiments described herein, a portion of the present invention is given by the following formula: TIFF2026517422000019.tif63170 (in the formula, R 6 , R 7 and R 8 R is represented as (independently C1-C6 alkyl or hydrogen). In some embodiments which can be combined with other embodiments described herein, 6 , R 7 , and R 8 Each is independently a C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments which can be combined with other embodiments described herein, R 6 , R 7 and R 8Each of these is methyl. In some embodiments which can be combined with other embodiments described herein, R 6 and R 7 is hydrogen, R 8 The C1-C6 alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments which can be combined with other embodiments described herein, R 6 and R 8 is hydrogen, R 7 The C1-C6 alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments which can be combined with other embodiments described herein, R 7 and R 8 is hydrogen, R 6 The C1-C6 alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments which can be combined with other embodiments described herein, R 6 is hydrogen, R 7 and R 8 R is independently a C1-C6 alkyl group, such as methyl, ethyl, propyl, butyl, pentyl, or hexyl. In some embodiments that can be combined with other embodiments described herein, R 7 is hydrogen, R 6 and R 8 R is independently a C1-C6 alkyl group, such as methyl, ethyl, propyl, butyl, pentyl, or hexyl. In some embodiments that can be combined with other embodiments described herein, R 8 is hydrogen, R 6 and R 7 These are independently C1-C6 alkyl groups, such as methyl, ethyl, propyl, butyl, pentyl, and hexyl.
[0037]
[0099] q is 0 or 1. In some embodiments which can be combined with other embodiments described herein, R 9 At least one of (multiple) is C1-C 16Alkyls include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl. Although not bound by theory, R 9 If the alkyl group is C5 or higher, the alkyl moiety can more easily penetrate the hydrophobic portion of the cell membrane (such as that of a viral cell), potentially accelerating the lysis of viral cells. Furthermore, the cationic nitrogen moiety can provide Coulomb interactions between the compound (such as the quaternary ammonium moiety) and the cell membrane surface (such as the phosphate portion of the phospholipid bilayer), further promoting the lysis of viral cells. In addition, the quaternary ammonium salt can confer sufficient hydrophilicity, thereby preventing the alkyl group bonded to the quaternary ammonium salt from being substantially embedded within the three-dimensional structure (for example, if the composition has a three-dimensional structure of a nanoworm or nanorod).
[0038]
[0100] In some embodiments that can be combined with other embodiments described herein, R 9 These are C1-C6 alkylenes, such as C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, etc. In some embodiments which can be combined with other embodiments described herein, R 9 R is an azole. In some embodiments which can be combined with other embodiments described herein, R 9 R is an oligomer of guanidine, for example, diguanidine. In some embodiments which can be combined with other embodiments described herein, 9 The following structure: This is phenyl represented by TIFF2026517422000020.tif56170.
[0039]
[0101] In some embodiments that can be combined with other embodiments described herein, R 9 The following structure: TIFF2026517422000021.tif62170(In the formula, x is an integer from 1 to 20, for example an integer of 10, and R* is hydrogen, -OH, or It is a polygalactose that has TIFF2026517422000022.tif20170.
[0040]
[0102] In some embodiments that can be combined with other embodiments described herein, R 9 R is a coumarin such as 7-hydroxycoumarin. In some embodiments which can be combined with other embodiments described herein, 9 R is a combination of azole and polygalactose (e.g., polygalactose-substituted azole). In some embodiments which can be combined with other embodiments described herein, 9 This is a combination of azole and coumarin (e.g., 3-azido-7-hydroxycoumarin), and is given by the following formula: It has TIFF2026517422000023.tif21170. For example, though not limited to, q is 1 and R 9 It is diguanidine.
[0041]
[0103] In at least one embodiment that can be combined with other embodiments described herein, the compound is of formula (I): Represented by TIFF2026517422000024.tif56170(I) or a pharmaceutically acceptable salt thereof, where n, m, and p are each independently integers from 1 to 100. Each example of q is independently an integer of 0 or 1. In some embodiments that can be combined with other embodiments described herein, n is an integer from 1 to 100, e.g., 20 to 50, 25 to 35, etc. In some embodiments, n is 30. In some embodiments that can be combined with other embodiments described herein, m is an integer from 1 to 100, e.g., 30 to 60, 35 to 55, etc. In some embodiments that can be combined with other embodiments described herein, n is 45. In some embodiments that can be combined with other embodiments described herein, p is an integer from 1 to 100, e.g., 45 to 55, 48 to 52, etc. In some embodiments that can be combined with other embodiments described herein, p is 50. In some embodiments that can be combined with other embodiments described herein, p is 52.
[0042]
[0104] In some embodiments that can be combined with other embodiments described herein, R of formula (I) 2 , R 3 and R 4 Each is independently hydrogen. In some embodiments which can be combined with other embodiments described herein, R 2 , R 3 , and R 4 Each is independently C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 and R 3 is hydrogen, R 4 is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 and R4 is hydrogen, R 3 is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 3 and R 4 is hydrogen, R 2 is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 is hydrogen, R 4 and R 3 C1-C is independent 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 3 is hydrogen, R 4 and R 2 C1-C is independent 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 And R4 is methyl, R 3 R is hydrogen. In some embodiments which can be combined with other embodiments described herein, R 4 is hydrogen, R 3 and R 2 C1-C is independent 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 and R 3 is methyl, and R 4 It is hydrogen.
[0043] In some embodiments that can be combined with other embodiments described herein, R of formula (I) 5 is a C6 aryl. In some embodiments, R 5 is phenyl. In some embodiments, phenyl is, for example, independently C1-C 10 The alkyl moiety can be substituted with 1, 2, 3, 4, or 5 moieties. This embodiment can be combined with any other embodiment described herein.
[0044]
[0106] In some embodiments that can be combined with other embodiments described herein, R of formula (I) 6 , R 7 , and R 8 Each is independently a C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments which can be combined with other embodiments described herein, R 6 , R 7 and R 8 Each of these is methyl. In some embodiments which can be combined with other embodiments described herein, R 6 and R 7 is hydrogen, R 8 The C1-C6 alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments which can be combined with other embodiments described herein, R 6 and R 8 is hydrogen, R 7 The C1-C6 alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments which can be combined with other embodiments described herein, R 7 and R 8 is hydrogen, R 6 The C1-C6 alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments which can be combined with other embodiments described herein, R 6 is hydrogen, R 7 and R 8is independently C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments that can be combined with other embodiments described herein, R 7 is hydrogen, R 6 and R 8 are independently C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. In some embodiments that can be combined with other embodiments described herein, R 8 is hydrogen, R 6 and R 7 are independently C1-C6 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, etc.
[0045]
[0107] In some embodiments that can be combined with other embodiments described herein, q of formula (I) is 0 or 1. In some embodiments that can be combined with other embodiments described herein, at least one of R 9 (s) is C1-C 16 alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments that can be combined with other embodiments described herein, R 9 is C1-C6 alkylene, such as C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, etc. In some embodiments that can be combined with other embodiments described herein, R 9 is azole. In some embodiments, R 9 is an oligomer of guanidine, such as diguanidine. In some embodiments that can be combined with other embodiments described herein, R 9 is the following structure: It is diguanidine represented by TIFF2026517422000025.tif56170.
[0046]
[0108] In some embodiments that can be combined with other embodiments described herein, R of formula (I)9 It is a polysaccharide such as polygalactose. In some embodiments that can be combined with other embodiments described herein, polygalactose has the following structure: TIFF2026517422000026.tif62170(where x is an integer from 1 to 20, for example, an integer of 10, and R* is hydrogen, -OH, or TIFF2026517422000027.tif20170).
[0047]
[0109] In some embodiments that can be combined with other embodiments described herein, R in formula (I) 9 is a coumarin such as 7-hydroxycoumarin. In some embodiments that can be combined with other embodiments described herein, R 9 is a combination of an azole and polygalactose (e.g., polygalactose-substituted azole). In some embodiments that can be combined with other embodiments described herein, R 9 is a combination of an azole and coumarin (e.g., 3-azido-7-hydroxycoumarin), and has the following formula: TIFF2026517422000028.tif21170. In at least one embodiment that can be combined with other embodiments described herein, q is 1 and R 9 is diguanidine.
[0048]
[0110] In some embodiments that can be combined with other embodiments described herein, R in formula (I) 10 and R 11 each independently is hydrogen or C1-C 20 alkyl, and at least one of R 10 or R 11 is C1-C 20 alkyl, such as methyl, ethyl, n-propyl or isopropyl. In at least one embodiment that can be combined with other embodiments described herein, at least one of R 10 or R 11 is isopropyl.
[0049]
[0111] In at least some embodiments that can be combined with other embodiments described herein, the nanostructure is of formula (II): It may be represented as TIFF2026517422000029.tif55170(II) or a pharmaceutically acceptable salt thereof. In some embodiments which can be combined with other embodiments described herein, each of r, s, t, u, m, and p in formula (II) is independently between 1 and 100. In some embodiments which can be combined with other embodiments described herein, r is an integer between 1 and 100, e.g., 5 to 40, 5 to 15, etc. In some embodiments which can be combined with other embodiments described herein, p is 11 or 12. In some embodiments which can be combined with other embodiments described herein, s is an integer between 1 and 100, e.g., 5 to 40, 5 to 15, etc. In some embodiments which can be combined with other embodiments described herein, p is 11 or 12. In some embodiments which can be combined with other embodiments described herein, t is an integer between 1 and 100, e.g., 1 to 20, 1 to 5, etc. In some embodiments which can be combined with other embodiments described herein, t is 3 or 4. In some embodiments that can be combined with other embodiments described herein, u is an integer from 1 to 100, for example, 1 to 20, 1 to 5, etc. In some embodiments that can be combined with other embodiments described herein, u is 3 or 4. In some embodiments that can be combined with other embodiments described herein, m is an integer from 1 to 100, for example, 25 to 60, 35 to 55, etc. In some embodiments that can be combined with other embodiments described herein, m is 45. In some embodiments that can be combined with other embodiments described herein, p is an integer from 1 to 100, for example, 30 to 70, 40 to 60, etc. In some embodiments that can be combined with other embodiments described herein, p is 50.
[0050]
[0112] In some embodiments which can be combined with other embodiments described herein, R of formula (II)1 is hydrogen or C1-C 20 It is alkyl. In some embodiments which can be combined with other embodiments described herein, R 1 R is butyl. In some embodiments which can be combined with other embodiments described herein, R 2 , R 3 and R 4 Each is independently hydrogen. In some embodiments which can be combined with other embodiments described herein, R 2 , R 3 , and R 4 Each is independently C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 and R 3 is hydrogen, R 4 is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 and R 4 is hydrogen, R 3 is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 3 and R 4 is hydrogen, R 2 is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 is hydrogen, R 3 and R 4 C1-C is independent 20Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 3 is hydrogen, R 2 and R 4 C1-C is independent 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 and R 4 is methyl, and R 3 R is hydrogen. In some embodiments which can be combined with other embodiments described herein, R 4 is hydrogen, R 2 and R 3 C1-C is independent 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 2 and R 3 is methyl, and R 4 It is hydrogen.
[0051]
[0113] In some embodiments which can be combined with other embodiments described herein, R of formula (II) 5 R is a C6 aryl compound. In some embodiments which can be combined with other embodiments described herein, R 5 is phenyl. In some embodiments that can be combined with other embodiments described herein, phenyl is independently C1-C 10 It can be substituted with one, two, three, four, or five parts, such as the alkyl portion.
[0052]
[0114] In some embodiments which can be combined with other embodiments described herein, R of formula (II) 6 , R 6’ , R 6’’ and R6’’’ Each of these is independently a C1-C6 alkyl group, such as methyl, ethyl, propyl, butyl, pentyl, or hexyl. In some embodiments that can be combined with other embodiments described herein, R 6 , R 6’ , R 6’’ and R 6’’’ Each of these is methyl. In some embodiments which can be combined with other embodiments described herein, R 7 , R 7’ , R 7’’ and R 7’’’ Each of these is independently a C1-C6 alkyl group, such as methyl, ethyl, propyl, butyl, pentyl, or hexyl. In some embodiments that can be combined with other embodiments described herein, R 7 , R 7’ , R 7’’ and R 7’’’ Each of these is methyl. In some embodiments which can be combined with other embodiments described herein, R 8 , R 8’ , R 8’’ and R 8’’’ Each of these is independently a C1-C6 alkyl group, such as methyl, ethyl, propyl, butyl, pentyl, or hexyl. In some embodiments that can be combined with other embodiments described herein, R 8 , R 8’ , R 8’’ and R 8’’’ Each of these is methyl.
[0053]
[0115] In some embodiments which can be combined with other embodiments described herein, R of formula (II) 10 and R 11 Each of them independently consists of hydrogen or C1-C 20 It is alkyl, R 10 or R 11 At least one of them is C1-C 20 Alkyl, for example, methyl, ethyl, n-propyl or isopropyl. In at least one embodiment which can be combined with other embodiments described herein, R10 or R 11 At least one of them is isopropyl.
[0054]
[0116] In some embodiments which can be combined with other embodiments described herein, R of formula (III) 12 R is an oligomer of guanidine, for example, diguanidine. In some embodiments which can be combined with other embodiments described herein, 12 The following structure: This is digunidine represented as TIFF2026517422000030.tif56170.
[0055]
[0117] In some embodiments which can be combined with other embodiments described herein, R of formula (III) 13 These are C1-C6 alkylenes, such as C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, etc. In some embodiments which can be combined with other embodiments described herein, R 13 R is a C3 alkylene. In some embodiments which can be combined with other embodiments described herein, R 13 This is a combination of azole and polygalactose (e.g., polygalactose-substituted azole), and is given by the following formula: TIFF2026517422000031.tif62170(In the formula, x is an integer from 1 to 20, for example, an integer of 10, and R* is hydrogen, -OH, or It has TIFF2026517422000032.tif20170). In some embodiments which can be combined with other embodiments described herein, R 13 This is a combination of azole and coumarin (e.g., 3-azido-7-hydroxycoumarin), and is given by the following formula: It has TIFF2026517422000033.tif19170.
[0056]
[0118] In some embodiments which can be combined with other embodiments described herein, R of formula (II) 14At least one of (multiple) is C1-C 16 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. In some embodiments which can be combined with other embodiments described herein, R 14 It is Octill.
[0057]
[0119] Although the aminoethyl acrylate units of formula (II) are shown as blocks, it should be understood that unsubstituted and substituted aminoethyl acrylate units can be randomly dispersed along the entire block of aminoethyl acrylate units, and the integer values of r, s, t, and u correspond not necessarily to blocks, but to the total number of each unit. Random arrangements of aminoethyl acrylate units and substituted aminoethyl acrylate units can be obtained by the functionalization methods described herein. Similarly, although the polyamide units of formula (II) are shown as blocks, they can instead exist as random units together with aminoethyl acrylate units. Random arrangements of polyamide units and aminoethyl acrylate units (substituted or unsubstituted) can be obtained by the polymerization methods described herein.
[0058] Nanostructure coating
[0120] Figure 1 is a schematic diagram showing a nanostructure such as a nanoworm 100 in a particular embodiment. The skeleton or core 110 of the nanoworm 100 comprises alkene units and macro CTA polymer units. The nanoworm 100 contains functional groups 120 of the macro CTA polymer units. Each functional group 120 is a component derived from a reversible addition-cleavage chain transfer (RAFT) agent and may be pre-functionalized or post-functionalized. In some examples, each functional group 120 is selected to modify the capture and deactivation / inactivation efficiency of the nanoworm 100 and / or to modify the responsiveness of the nanoworm 100 (e.g., temperature, pH, salt concentration, light, and / or combinations thereof).
[0059] Three-dimensional structure of a compound or composition
[0121] The compounds or compositions of this disclosure (e.g., two or more different nanostructures) may have a three-dimensional structure that is a nanoworm or a nanorod. Nanorods may have an aspect ratio (i.e., length:width ratio) of about 10:1 to about 1000:1, for example, about 10:1 to about 100:1, for example, about 25:1 to about 75:1. Nanorods may have a diameter of about 10 nm to about 20 nm and a length of about 100 nm to about 10 microns, for example, about 1 micron to about 2 microns. Nanoworms may have an aspect ratio greater than about 1000:1.
[0060]
[0122] The compounds and compositions of this disclosure may also have spherical, vesicular, donut-shaped, or lamellar sheet-like three-dimensional structures. The three-dimensional structures of the compositions of this disclosure are stable in water for extended periods (e.g., nanoworms stable at room temperature for more than one year) and can be freeze-dried and rehydrated without structural rearrangement. For example, a nanoworm solution can be freeze-dried to obtain a dry powder. This freeze-dried product can be rehydrated in Milli-Q water at -8% by weight for 2 hours. The freeze-dryability of the compositions of this disclosure enables stable transport of the compositions of this disclosure.
[0061] Nanoworm synthesis
[0123] A method for forming nanostructures is provided. In some embodiments, polymer nanostructures having a nanoworm morphology can be directly produced in water using emulsion polymerization. This method involves introducing a styrene monomer into a reactor to form a mixture, comprising (1) a first polymer having N-alkylacrylamide units and (2) a second polymer having N,N-(dialkylamino)(divalent alkyl)alkyl acrylate units and N-alkylacrylamide units. The polystyrene block can provide a high glass transition temperature (Tg) component (the Tg of 100% polystyrene is approximately 100°C). A high Tg results in the stability of the nanostructure at body temperature. Furthermore, the poly(N-isopropylacrylamide) (poly-NIPAM) block can provide three-dimensional conformation of nanoworms (or nanorods) under aqueous conditions, for example, in an aqueous solution containing sodium dodecyl sulfate (SDS). In some embodiments, the introduction of styrene monomers comprising a first polymer having N-alkylacrylamide units and a second polymer having N,N-(dialkylamino)(divalent alkyl)alkyl acrylate units and N-alkylacrylamide units is carried out at a temperature of about -10°C to about 10°C. The first polymer comprises (1) an N-isopropylacrylamide unit as the N-alkylacrylamide unit, and (2) a portion represented by the following formula: TIFF2026517422000034.tif19170 (in the formula, R 1 (is alkyl), and the part represented by formula (3): TIFF2026517422000035.tif27170 (in the formula, R 2 is alkyl (branched or linear, substituted or unsubstituted), R 3 and R 4 These can independently consist of hydrogen or alkyl (branched or linear, substituted or unsubstituted).
[0062]
[0124] In some embodiments, the first polymer does not include N,N-(dialkylamino)(divalent alkyl)alkyl acrylate units. The second polymer includes (1) N,N-(dialkylamino)(divalent alkyl)alkyl acrylate units as N,N-(dimethylamino)ethyl methacrylate units, N-alkylacrylamide units as N-isopropylacrylamide units, a moiety represented by the formula: TIFF2026517422000036.tif19170(where R 1 is alkyl), and (2) a moiety represented by the formula: TIFF2026517422000037.tif30170[where R 2 is alkyl (branched or straight-chain, substituted or unsubstituted), and R 3 and R 4 are independently hydrogen or alkyl (branched or straight-chain, substituted or unsubstituted)].
[0063]
[0125] This method includes introducing an initiator compound into the above mixture to form a second mixture having the nanostructure. The initiator compound is a peroxide, hydroperoxide or azo initiator. In some examples, the initiator is azobisisobutyronitrile.
[0064]
[0126] As shown in Figure 1B, the nanostructure can be bound to various functional groups, including hydrophobic octane (O), diguanidine (DG), a fluorescent probe (C) (e.g., coumarin), and a polysaccharide, such as polygalactose (S). Binding to the highly glycosylated spike S protein was targeted via (i) strong polyvalent binding with polygalactose, and (ii) electrostatic interactions between the negatively charged viral particle and the positively charged guanidine and N,N-(dimethylamino)ethyl methacrylate (DMAEMA) groups. The bound octane group facilitates the rupture of the viral membrane, in which case the viral mRNA is degraded by the diguanidine group or electrostatically captured by the polymer coating. The polymer nanostructures described herein can be coated onto surfaces, including surgical masks, to readily inactivate sense RNA viruses, such as influenza A virus, ancestral isolates of SARS-CoV-2, alpha mutants, and omicron mutants.
[0065]
[0127] In certain embodiments, the nanostructure comprises a copolymer of macro-chain transfer agent (macro-CTA) polymer units and alkene units. The macro-CTA polymer is a polymer formed by RAFT using a RAFT agent in the polymerization of one or more ethylenically unsaturated monomers.
[0066]
[0128] In some cases, two types of macrochain transfer RAFT poly(N-isopropylacrylamide) (PNIPAM) agents can be produced from one non-functional RAFT agent. Emulsification polymerization using two types of macrochain transfer agents in the presence of styrene (e.g., initiated with azobisisobutyronitrile (AIBN) in a 500 mL reactor at 70°C) can produce spherical particles consisting of two block copolymers, macro-CTA A and B, with a polymer content of approximately 8 wt% in water. After adding a small amount of plasticizer for polystyrene, the spherical nanoparticles transform into nanoworms upon cooling to room temperature. This synthesis process is called temperature-directed morphogenesis (TDMT) and can be used to produce a wide range of polymer nanoparticles, including worm-like, rod-like, vesicle-like, toroidal, tadpole-like, stacked toroidal rattle-type nanoparticles (nanorattle), and other forms, or combinations thereof.
[0067]
[0129] Subsequently, these polymer nanoworms can be bonded to functional groups (O, G) via quaternization, dialyzed, freeze-dried, and rehydrated with water to produce a 1.5 wt% polymer / water dispersion. Then, the polymer nanoworms can be bonded to functional groups (S and C) via a copper-catalyzed azidoalkyne cycloaddition (CuAAC) reaction. A combination of CuSO4 and sodium ascorbate can be used for the CuAAC. These samples can then be dialyzed, freeze-dried, and rehydrated with water to produce a 1.5 wt% polymer / water dispersion. Finally, this polymer (NWS) ,S,O,C,G The dispersion can be coated onto the surface with 1 to 5 sprays. The amount of polymer per unit area can be determined by measuring the dry weight of the polymer on the glass slide surface using a microbalance.
[0068] Synthesis of nanorods
[0130] Nanorods can be obtained by temperature-directed morphogenesis (TDTM) and ultrasonic cutting of nanoworms. In at least one embodiment, 6 mL of a latex solution of nanoworms can be transferred to two warmed vials (3 mL each) containing 60 μL of toluene in each vial. These vials can then be sealed and shaken. The suspensions in each vial can be cooled to 23°C. These suspensions can be cooled from 70°C to 15°C in approximately 30 minutes. The formation of worm-like nanostructures can be confirmed by characterizing these nanostructures with a transmission electron microscope (TEM). To form rods, the worms are diluted with 10 mL of Milli-Q water and cut at 35% amplitude for 3 minutes in an ice bath using an ultrasonic probe (3 seconds on and 2 seconds off as one pulse cycle) (Sonics & Materials VC-750 system, 3 mm tapered microtip). After ultrasonic cutting, the nanostructures can be characterized again by TEM to confirm rod formation.
[0069]
[0131] Ultrasonic cutting of nanoworms into nanorods can also be performed by applying probe-type ultrasound at different pulse cycles (15 seconds on and 10 seconds off as one pulse cycle): (B) 12 cycles (3 minutes), (C) 36 cycles (9 minutes), and (D) 48 cycles (12 minutes).
[0070]
[0132] In at least one embodiment, when a nanoworm or nanorod composition of the present disclosure is heated to a temperature above the lower critical dissolution temperature (LCST) of the PNIPAM block (e.g., about 37°C), a gel is obtained that can be redissociated into a sol upon cooling, and the process is reversible. Nanoworms can form a gel in an aqueous solution when the minimum weight fraction of nanoworms relative to the total volume of the aqueous solution is about 0.1% to about 10% (e.g., about 1% to about 8%). Nanorods can form a gel when the minimum weight fraction of nanorods in an aqueous solution is about 2% to about 16%. Nanostructured samples have a distribution of nanostructure length (aspect ratio), and gel formation may depend on the aspect ratio present in the nanostructured sample. Although not bound by theory, gels are advantageous because they can dissociate with increasing temperature (e.g., from room temperature to the body temperature of an object such as a human). As a result, the three-dimensional structure of the nanoworm can dissociate and travel through the bloodstream.
[0071]
[0133] The weight percentage of nanorods in water capable of forming a gel at 37°C can be measured by the following method: Generally, lyophilized nanorods (e.g., 20 mg) can be redispersed in Milli-Q water at 25°C by vortexing in a 1.5 ml Eppendorf tube at 25°C with 30% by weight of nanorods. The tube is then capped and immersed in a water bath at 37°C for 2 minutes. The tube is then inverted underwater, and gel formation can be observed. Gel formation is defined as the absence of fluid flow within 30 seconds. The weight percentage can be reduced by adding more Milli-Q water and vortexing. Gel formation can then be confirmed again. The minimum weight percentage of nanorods (e.g., in water) that forms a gel at 37°C is defined as the gel-forming weight percentage.
[0072] Methods for depositing compounds and compositions
[0134] The compounds and compositions of this disclosure can be deposited on the surface of an object by any suitable deposition method. The surface of the object may be any suitable surface of any suitable object. The surface may be porous or nonporous. The deposition method may include one or more of the following: coating, dipping, spraying, marking, taping, brush coating, spin coating, roll coating, and doctor blade coating. Before deposition, the compounds or compositions of this disclosure may be diluted with a polar solvent, a protic solvent, an aprotic solvent, for example, water. After deposition, the solvent evaporates at room temperature, forming a compound / composition layer on the object.
[0073]
[0135] In at least one embodiment, the object is an interior surface of an aircraft / spacecraft / ship or an air filter surface of an aircraft / spacecraft / ship (such as the surface of an air conditioning system or filtration system). The object is an interior floor of an aircraft, a seat surface (including but not limited to armrests / headrests), a seat belt buckle, a seat pocket, a tray table, an overhead luggage rack surface, a ceiling surface, a trim surface, a screen surface, a window surface, a door surface, and / or a door handle surface.
[0074]
[0136] In at least one embodiment, the compound or composition of the Disclosure is applied to the surface of an object for about 1 second to about 10 minutes, for example, about 30 seconds to about 2 minutes (e.g., by spraying, depositing, printing, etc.). In at least one embodiment, the compound or composition is applied to the surface of an object in an amount of about 1 mL to about 25 kL, for example, about 100 L to about 1 kL (e.g., by spraying). The compound or composition of the Disclosure can be applied so that the surface appears wet due to the solvent of the composition.
[0075]
[0137] Compounds or compositions of the present disclosure placed on an object can prevent, reduce, and / or eliminate the presence of bacteria and viruses (such as SARS-CoV-2), thereby preventing, reducing, and / or eliminating contact between such bacteria and viruses and humans. Compounds or compositions of the present disclosure bind to bacteria and / or viruses as described herein, in which case conformational changes occur in nanoworms and / or nanorods, causing the bacterial and / or viral membranes to rupture and resulting in the inactivation of bacteria and / or viruses. Inactivation reduces the amount of contact between bacteria and / or viruses and humans.
[0076]
[0138] The composition may have any suitable pH, for example, between approximately 6.5 and approximately 7.4. For example, a pH of approximately 6.5 mimics the pH of mucosal droplets. The composition may have a pH that is interactable with humans and is within the human biological tolerance range. Therefore, the composition is suitable for administration to biological membranes (such as human mucosa). While we do not wish to be bound by theory, the pH of the composition may enhance its antibacterial or antiviral capabilities, and a pH that mimics the pH of mucosal droplets may help reduce the presence of bacteria or viruses.
[0077]
[0139] Compositions containing the nanostructures (e.g., nanorods or nanoworms) of this disclosure are advantageous for deposition on surfaces. This is because, for example, antimicrobial and antiviral compounds can be applied as a single layer while maintaining the effectiveness of both compounds. Applying the nanostructured composition as a single layer also reduces the cost and time required to apply the compounds to the surface compared to applying two or more layers. Using an aqueous solution ensures end-user safety and results in reduced application time and cost to the surface. Alternatively, in some examples, thicker layers and / or multiple layers can be applied. In some examples, based on the desired level of antimicrobial or antiviral protection, the surface is reapplied or supplemented with one or more additional layers of the nanostructured composition at once after the initial application (one or more layers).
[0078]
[0140] In some embodiments, the method involves impregnating a cloth (woven or nonwoven) or fiber. In some embodiments, a method for arranging nanostructures on a surface is described herein. In some embodiments, the method involves arranging a layer of a solution containing the nanostructures on a surface. The nanostructures include compounds or salts thereof.
[0079]
[0141] The surface to be treated with the coating may be any suitable surface or any suitable object that can be coated or impregnated. In some non-limiting embodiments, the object is a mask, and the surface is an interior part of an aircraft fuselage or other suitable surface. In some examples, the surface is the surface (interior or exterior) of an aircraft, ship, train, terminal building (such as a bus, train, or airport), or spacecraft.
[0080]
[0142] In some embodiments, the emulsion or solution has a concentration of the nanostructures of about 0.5% to about 3% by weight relative to the total volume of the solution.
[0081] Method of use as a pharmaceutical product
[0143] In some embodiments, the Disclosure further provides a method for treating a condition in a subject who has or is susceptible to a certain condition by administering a therapeutically effective amount of one or more compounds or compositions of the Disclosure to the subject. In some embodiments, the treatment is prophylactic. In other embodiments, the treatment is palliative. In other embodiments, the treatment is regenerative.
[0082]
[0144] A method for treating a certain condition may involve administering a therapeutically effective amount of a nanostructure or a pharmaceutically acceptable salt thereof (or a composition having a nanostructure or a pharmaceutically acceptable salt thereof) to a target.
[0083]
[0145] Methods for treating a certain condition are described herein. In some embodiments, the methods include administering a therapeutically effective amount of nanostructures to a target.
[0084]
[0146] The coatings described herein are scalable and can be applied directly to surfaces as aqueous solutions to act as effective virucidal agents that render concerning SARS-CoV-2 variants non-infectious. The design of polygalactose (e.g., about 2 to about 20 galactose units) bound to polymer nanostructures and the potential specific binding interactions with highly glycosylated SARS-CoV-2 provide a binding motif that is independent of the mutations found in viral variants and viral spike-attached glycoproteins. In some embodiments, the polygalactose has more than 20 galactose units, e.g., up to about 1000 galactose units. The combination of polygalactose binding and octane moieties, along with the responsiveness of the nanostructures that mechanically attach to viral particles, destroy them, and render them non-infectious, is at work. While not bound by theory, the SARS-CoV-2 viral RNA genome would likely be degraded by the digunidine group or electrostatically captured by the cationic group bound to the polymer and subsequently biodegraded. After the virus interacted with a polymer-coated surface, the viral RNA genome was found to be undetectable, demonstrating the polymer's complete antiviral activity. This polymer coating is thought to inactivate newly emerging, concerning SARS-CoV-2 variants while simultaneously retaining the ability to be redesigned through functionalization to target other viruses. Ultimately, the polymer was confirmed to be non-toxic upon oral ingestion by rats, and showed little to no skin sensitization when applied to mouse skin. This suggests its potential for safe use as a component in personal protective equipment and high-contact surfaces that come into contact with the skin. The nanostructured composition can also be administered to subjects as a therapeutic treatment.
[0085] 1.Conditions
[0147] Conditions that can be treated in accordance with this disclosure include, but are not limited to, conditions caused by toxins (such as antigens) and inflammatory diseases such as septic shock. Conditions that can be treated in accordance with this disclosure include, but are not limited to, viral infections, bacterial infections, chronic inflammatory diseases, acute inflammatory diseases, and cancer. In some embodiments, conditions that are eligible for treatment include bacterial infections, viral infections, or conditions associated with cancer immunotherapy. Cancer immunotherapy also includes cervical cancer, such as that caused by human papillomavirus infection of the cervix.
[0086]
[0148] Viral infections include Ebola hemorrhagic fever, influenza, SARS (such as SARS-CoV-2), norovirus (gastroenteritis), or Zika fever. Viral infections also include viral respiratory infections (e.g., infections of the nose, throat, upper respiratory tract, lungs, etc.) such as pneumonia, laryngotracheobronchitis, and bronchiolitis. Viral infections include viral gastroenteritis, such as gastroenteritis caused by norovirus or rotavirus. Viral infections include viral liver infections, such as hepatitis. Viral infections include viral neurological infections, such as encephalitis caused by rabies or West Nile virus. Viral infections include warts and / or infections caused by human papillomavirus (HPV). Viral infections also include cancer-causing infections, such as infections caused by Epstein-Barr virus, hepatitis B, hepatitis C, herpesvirus 8, or human papillomavirus. Symptoms of viral infections include fever, muscle pain, cough, sneezing, runny nose, headache, chills, diarrhea, vomiting, rash, or weakness.
[0087]
[0149] Bacterial infections include pneumonia, meningitis, food poisoning, and those caused by Staphylococcus or Streptococcus, as well as cellulitis, folliculitis, impetigo, and abscesses. Bacterial infections (e.g., food poisoning) include infections caused by Escherichia coli, Campylobacter jejuni, Clostridium botulinum, Listeria monocytogenes, Salmonella, and Vibrio. Bacterial infections also include bacterial meningitis, otitis media, urinary tract infections, and respiratory infections such as pharyngitis, bronchitis, sinusitis, and pneumonia. Symptoms of bacterial infections include nausea, vomiting, diarrhea, fever, chills, and abdominal pain.
[0088]
[0150] In some embodiments, the methods described herein are used to treat patients having disorders resulting from the production, stability, secretion, and post-translational processing of dysregulated cytokines, enzymes, and / or inflammatory mediators. Examples of cytokines that may cause dysregulation include interleukins 1, 2, 6, 8, 10, 12, 17, 22, 23, tumor necrosis factor alpha, interferon alpha, beta, and gamma. Examples of inflammatory mediators that may cause dysregulation include nitric oxide, prostaglandins, and leukotrienes. Examples of enzymes include cyclooxygenase, nitric oxide synthase, and matrix metalloproteinases.
[0089]
[0151] Examples of inflammatory conditions related to the technology of the present invention include, but are not limited to, sepsis, septic shock, endotoxin shock, exotoxin-induced toxic shock, Gram-negative sepsis, and toxic shock syndrome. Inflammatory conditions may include those experienced by immunocompromised individuals, as well as "superbugs," including bacterial and viral strains resistant to current therapeutic agents.
[0090] 2. Target
[0152] Suitable subjects to be treated in accordance with this disclosure include mammalian subjects. Mammals as disclosed include, but are not limited to, humans, dogs, cattle, cattle, goats, horses, sheep, pigs, rodents, rabbits, and primates, and also include mammals in utero. Subjects may be of any sex and at any stage of development.
[0091] 3. Dosage and Administration
[0153] The compounds or compositions disclosed herein can be administered to a subject in a therapeutically effective amount.
[0092]
[0154] The compounds or compositions of this disclosure can be administered via any suitable route, in the form of a pharmaceutical composition adapted to such route, in a dose effective for the intended treatment. The effective dose is typically in the range of about 0.001 mg to about 100 mg per kg of body weight per day, for example, about 0.01 mg to about 30 mg / kg / day, and is administered as a single dose or in divided doses. Depending on age, species, and the condition being treated, doses below the lower limit of this range may be appropriate. In other cases, higher doses may be administered without side effects. Higher doses may also be divided into multiple small doses for administration throughout the day.
[0093] Pharmaceutical composition
[0155] For the treatment of the above condition, the compounds described herein may be administered as follows:
[0094] Oral administration
[0156] The compounds or compositions of this disclosure can be administered orally, including by swallowing, so that the compounds enter the gastrointestinal tract, or they can be absorbed directly into the bloodstream from the mouth (e.g., buccal or sublingual administration).
[0095]
[0157] Compositions suitable for oral administration include solid formulations such as tablets, lozenges, and capsules, which may contain liquids, gels, or powders. Compositions for oral administration can be formulated as immediate-release or modified-release (including delayed-release or sustained-release) formulations and may optionally be coated with an enteric coating.
[0096]
[0158] Liquid formulations include solutions, syrups, and suspensions, which can be encapsulated in soft capsules or hard capsules. These formulations may contain pharmaceutically acceptable carriers, such as water, ethanol, polyethylene glycol, cellulose, or oil. They may also contain one or more emulsifiers and / or suspending agents.
[0097]
[0159] In tablet dosage forms, the amount of compound present can be about 0.05% to about 95% by weight of the dosage form, for example, about 2% to about 50% by weight. Furthermore, tablets may contain about 0.5% to about 35% by weight of a disintegrant, for example, about 2% to about 25% by weight. Examples of disintegrants include methylcellulose, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, polyvinylpyrrolidone, hydroxypropylcellulose, or starch.
[0098]
[0160] Lubricants suitable for use in tablets may be present in an amount of approximately 0.1% to 5% by weight. Examples of lubricants include calcium stearate, zinc stearate, magnesium stearate, or sodium stearyl fumarate.
[0099]
[0161] Suitable binders for use in tablets include gelatin, polyethylene glycol, sugar, gum, starch, and hydroxypropyl cellulose. Suitable diluents for use in tablets include mannitol, xylitol, lactose, dextrose, sucrose, sorbitol, and starch.
[0100]
[0162] Suitable surfactants and lubricants for use in tablets may be present in amounts ranging from approximately 0.1% to approximately 3% by weight, based on the weight of the surfactant in the tablet. Examples of surfactants and lubricants include polysorbate 80, sodium dodecyl sulfate, talc, or silicon dioxide.
[0101] Parenteral administration
[0163] The compounds and compositions of this disclosure can be administered directly into the bloodstream, muscles, or internal organs. Suitable methods for parenteral administration may include intravenous, intramuscular, subcutaneous, intra-arterial, intraperitoneal, intrathecal, or intracranial administration. Suitable devices for parenteral administration include syringes (including needle-equipped and needle-free syringes) and infusion methods.
[0102]
[0164] Compositions for parenteral administration can be formulated as immediate-release or modified-release (including delayed-release or sustained-release) formulations.
[0103]
[0165] Most parenteral formulations are aqueous solutions containing additives such as salts, buffers, and carbohydrates. Parenteral formulations may also include non-aqueous or organic solutions containing additives such as salts, buffers, and carbohydrates.
[0104]
[0166] Parenteral formulations can also be prepared in dehydrated form (e.g., by lyophilization) or as sterile, non-aqueous solutions. These formulations may contain water. Solubility enhancers can also be used in the preparation of parenteral solutions.
[0105] Local administration
[0167] The compounds and compositions of this disclosure can be administered topically or transdermally to the skin. Formulations for topical administration include lotions, solutions, creams, gels, hydrogels, ointments, foams, implants, and patches. Pharmaceutically acceptable carriers for topical formulations include water, alcohol, mineral oil, glycerin, and polyethylene glycol. Topical administration can be performed by electroporation, iontophoresis, or phonophoresis.
[0106]
[0168] Compositions for topical administration can be formulated as immediate-release or modified-release (including delayed-release or sustained-release) formulations.
[0107] Combination and combination therapy
[0169] The compounds and compositions of this disclosure can be used alone or in combination with other pharmaceutically active compounds to treat the conditions described above. The compounds / compositions of this disclosure and other pharmaceutically active compounds can be administered simultaneously (in the same dosage form or in separate dosage forms) or sequentially. Accordingly, in at least one embodiment, this disclosure includes a method for treating a certain condition by administering to a subject one or more of the compounds of this disclosure and one or more additional different pharmaceutically active compounds, each in a therapeutically effective dose.
[0108]
[0170] In another embodiment, a pharmaceutical composition is provided comprising one or more compounds of the present disclosure, one or more additional pharmaceutically active compounds, and a pharmaceutically acceptable carrier.
[0109]
[0171] In another embodiment, one or more additional different pharmaceutically active compounds are one or more anti-inflammatory agents, anti-atherosclerotic agents, immunosuppressants, immunomodulators, cell division inhibitors, antiproliferative agents, angiogenesis inhibitors, kinase inhibitors, cytokine blockers, or inhibitors of cell adhesion molecules.
[0110]
[0172] Furthermore, the compounds and compositions of this disclosure may be used in combination with other therapeutic agents selected for their therapeutic value for the condition being treated. Generally, in embodiments in which combination therapy is employed, the compounds and compositions described herein and other agents do not need to be administered in the same pharmaceutical composition and may be administered via different routes of choice due to their differing physical and chemical properties. The initial dose is usually administered according to an established protocol, and thereafter, the dose, mode of administration, and timing of administration may be modified based on the observed effects. In certain cases, it is appropriate to administer the compounds of this disclosure in combination with another different therapeutic agent. For example, if one of the side effects experienced by a patient administered with a compound of this disclosure is a rash, it is appropriate to administer an antihistamine in combination with the initial therapeutic agent. Alternatively, for example, the therapeutic effect of the compounds of this disclosure may be enhanced by the administration of another therapeutic agent (including a therapeutic regimen) that has therapeutic benefits. Regardless of the disease, disorder, or condition being treated, the overall benefit experienced by the patient may simply be the sum of the two therapeutic agents, or the patient may experience a synergistic benefit.
[0111]
[0173] When therapeutic agents are used in combination, the therapeutically effective dose varies. Methods for experimentally determining the therapeutically effective doses of drugs and other agents used in combination therapy regimens are documented as methodologies. Combination therapy also includes periodic treatments that are initiated and discontinued at various times to support the patient's clinical management. In any case, multiple therapeutic agents (one of which is a compound of this disclosure) are administered in any order or concurrently. If administered concurrently, the multiple therapeutic agents are provided, at their discretion, in a single unified form or in multiple forms (for example, as a single tablet or as two separate tablets).
[0112]
[0174] In some embodiments, one or both therapeutic agents are administered multiple times. If not simultaneously, the timing between multiple administrations is arbitrarily varied, ranging from 0 weeks to less than 12 weeks.
[0113]
[0175] Furthermore, the combination methods, compositions, and formulations are not limited to the use of only two drugs, but also envision the use of multiple therapeutic combinations. The administration regimen for treating, preventing, or improving the condition for which relief is sought may be selectively modified depending on various factors. These factors include the disorder the subject is suffering from, as well as the subject's age, weight, sex, diet, and medical condition. Therefore, the medication regimen actually used may vary considerably in some embodiments and may deviate from the medication regimens defined herein.
[0114]
[0176] The agents constituting the combination therapies disclosed herein may optionally be in combination dosage forms or individual dosage forms intended for substantially simultaneous administration. The agents constituting the combination therapy may optionally be administered sequentially, with one of the agents being administered in a two-step regimen. The two-step regimen may optionally require sequential administration of the active agents or intervald administration of separate active agents. The time between multiple administration steps may range from several minutes to several hours, depending on the characteristics of each agent, such as their potency, solubility, bioavailability, plasma half-life, and kinetic profile. Diurnal variation in target molecule concentrations may optionally be used to determine the optimal administration interval.
[0115]
[0177] The compounds of this disclosure or compositions comprising the compounds of this disclosure may be used (e.g., administered) in combination with agents of the following drug classes: NSAIDs, immunosuppressants, immunomodulators, cell division inhibitors, antiproliferative agents, angiogenesis inhibitors, biological agents, steroids, vitamin D3 analogs, retinoids, other kinase inhibitors, cytokine blockers, corticosteroids, and inhibitors of cell adhesion molecules. If a subject has or is at risk of developing atherosclerosis or a condition associated with atherosclerosis, the compounds or compositions of this disclosure may be used optionally, in any combination, with one or more agents or methods for treating atherosclerosis or a condition associated with atherosclerosis. Examples of therapeutic agents / treatments for atherosclerosis or conditions associated with atherosclerosis include, but are not limited to, torcetrapib, aspirin, niacin, HMG-CoA reductase inhibitors (e.g., atorvastatin, fluvastatin, lovastatin, pravastatin, rosuvastatin, and simvastatin), cholesveram, cholestyramine, colestipol, gemfibrozil, probucol, and clofibrate.
[0116]
[0178] If the subject is exhibiting or at risk of exhibiting an inflammatory condition, the compounds or compositions of this disclosure may be optionally used in combination with one or more agents or methods for treating the inflammatory condition. Examples of therapeutic agents / methods for treating autoimmune diseases and / or inflammatory conditions include, but are not limited to, corticosteroids, nonsteroidal anti-inflammatory drugs (NSAIDs) (e.g., ibuprofen, naproxen, acetaminophen, aspirin, fenoprofen (Nalphon®), flurbiprofen (Anseid®), ketoprofen, oxaprozin (DAYPRO®), diclofenac sodium (Volta) Len®, diclofenac potassium (Cataphram®), etodolac (Rodin®), indomethacin (Indocine®), ketorolac (TORADOL®), sulindac (Clinoryl®), tolmetin (Trectin®), meclofenamic acid (MECLOMEN®), mefenamic acid (PONSTEL®), nabumetone (RELAFEN®), piroxicam (Felden®), Cox-2 inhibitors (e.g., celecoxib (CELEBREX®)), immunosuppressants (e.g., methotrexate (Rheumatrex®), leflunomide (ARAVA®), azathioprine (Imuran®), cyclosporine (Neoral®, Sandimmune®), tacrolimus and cyclophosphamide (CYTOXAN®), CD20 blockers (rituximab®), tumor necrosis factor (TNF) blockers (e.g., etanercept) (Enbrel®), infliximab (Remicade®), adalimumab (Humira®), abatacept (CTLA4-Ig), interleukin-1 receptor antagonists (e.g., anakinra (Kineret®)), interleukin-6 inhibitors (e.g., Actemra®), interleukin-17 inhibitors (e.g., AIN457), Janus kinase inhibitors (e.g., Tasocitinib), SYK inhibitors (e.g., R788), chloroquine and its derivatives.
[0117]
[0179] For use in cancer and neoplastic diseases, the compounds or compositions of this disclosure are optionally used in combination with one or more of the following drug classes: where the anticancer agent is an EGFR kinase inhibitor, MEK inhibitor, VEGFR inhibitor, anti-VEGFR2 antibody, KDR antibody, AKT inhibitor, PDK-1 inhibitor, PI3K inhibitor, c-kit / Kdr tyrosine kinase inhibitor, Bcr-Abl tyrosine kinase inhibitor, VEGFR2 inhibitor, PDGFR-beta inhibitor, KIT inhibitor, Flt3 tyrosine kinase inhibitor, PDGF receptor family inhibitor, Flt3 tyrosine kinase inhibitor, RET tyrosine kinase receptor family inhibitor, VEGF-3 receptor antagonist, Raf protein kinase family inhibitor, angiogenesis inhibitor, Erb2 inhibitor, mTOR inhibitor, IGF-1R antibody, NFκB inhibitor, proteosome inhibitor, chemotherapeutic agent, or hypoglycemic agent. [Examples]
[0118]
[0180] Reagents: Unless otherwise noted, all chemicals were used as received. The solvents used were either HPLC or AR grade and included: dichloromethane (DCM, Aldrich AR grade), DMSO (Aldrich, 99.9%), n-hexane (Emsure, ACS), chloroform (Emsure, ACS), methanol (Merck, Emsure, ACS), acetonitrile (LiChrosolv, hypergrade for LC-MS), petroleum spirits (BR 40°C~60°C, Univar, AR), toluene (Merck, EMSURE ACS for analysis, ISO, Reag.Ph Eur), ethyl acetate (ChemSupply, AR), ethanol (ChemSupply, AR), N,N-dimethylformamide (DMF:Labscan, AR grade), and N,N-dimethylacetamide (Aldrich, >99%).Activated basic alumina (Aldrich: Brockmann I, standard grade, approx. 150 mesh, 58 Å), silica gel (Aldrich, 230-400 mesh, 60 Å), magnesium sulfate (anhydrous, Aldrich), Milli-Q water (Biolab, 18.2 MΩm), sodium dodecyl sulfate (SDS, Aldrich, 99%), 1-butanethiol (Aldrich, 99%), D-(+)-galactose (Aldrich, ≥99%), propargyl bromide solution (Aldrich, 80% by weight in toluene, containing 0.3% magnesium oxide as a stabilizer), lithium chloride (Aldrich, 99%), tripotassium phosphate (Aldrich, ≥98%), potassium hydroxide (Aldrich), 3-chloropropylamine hydrochloride (Aldrich, 98%), triethylamine (Aldrich, ≥99.5%), acryloyl chloride (Mel) Use the following ingredients as obtained: (Stabilized with phenothiazine), sodium bicarbonate (Aldrich, 99.5%), sodium azide (Aldrich, ≥99.5%), hydrochloric acid (36%, Ajax, AR), sulfuric acid (Aldrich, 98%), trifluoroacetic acid (Merck, >99%), carbon disulfide (Aldrich, >99.9%), methyl-2-bromopropionate (MBP, Aldrich, 98%), 2-ethyl-2-thiopsoid urea hydrobromide (Aldrich, 98%), iodooctane (Aldrich, 98%), copper(II) sulfate (Aldrich, 99%), anhydrous copper(II) sulfate powder (Aldrich, ≥99.99% based on trace metals), copper powder (Aldrich, <425 μm, based on 99.5% based on trace metals), and L-ascorbic acid (Aldrich, 99%).
[0119]
[0181] Monomers, initiators, and ligands: N-isopropylacrylamide (NIPAM, Aldrich, 97%) and N,N-(dimethylamino)ethyl methacrylate (DMAEMA, Aldrich, 98%) were dissolved in ethanol with activated basic alumina, filtered, and used directly for the synthesis of macro-chain transfer agents (macro-CTA). Styrene (STY, Aldrich, >99%) was passed through a basic alumina column to remove inhibitors. Azobisisobutyronitrile (AIBN, Riedel-de Haan) was recrystallized twice from methanol before use. Tris(2-dimethylamino)ethyl)amine (Me6TREN), 1 Cu(II)Br2 / Me6TREN complex, 2 3-azido-7-hydroxycoumarin azide (coumarin azide) 3 were synthesized according to literature procedures.
[0120]
[0182] RAFT agent: Methyl 2-butylthiocarbonothiolthio)propanoate (MCEBTTC) RAFT agent was synthesized according to the literature procedure.
[0121]
[0183] Nuclear Magnetic Resonance (NMR): All NMR spectra were recorded using either a Bruker DRX400 or a 500 MHz spectrometer with an external lock (CDCl3, DMSO-d6, or D2O).
[0122]
[0184] Size exclusion chromatography (SEC) and triple detection size exclusion chromatography (TD-SEC): The molecular weight distribution of polymers was analyzed using a Polymer Laboratories GPC50 Plus equipped with a differential refractive index detector. The absolute molecular weight of polymers was measured using a Polymer Laboratories GPC50 Plus equipped with a dual-angle laser light scattering detector, a viscometer, and a differential refractive index detector. 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 performed using two PLGel Mixed B (7.8 × 300 mm) SEC columns connected in series and maintained at a constant temperature of 50°C. InfinityLab EasiVial polystyrene standards were used for SEC column calibration. Samples of known concentration were freshly prepared with 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 the Polymer Laboratories Multi Cirrus software based on the quantitative mass recovery technique.
[0123]
[0185] Dynamic Light Scattering (DLS): Particle size and zeta potential were measured by DLS. DLS was performed using a Malvern Zetasizer Nano series running DTS software and a 4mW He-Ne laser at 633nm. The analysis was performed at a constant angle of 173° and temperature of 25°C. Number-mean hydrodynamic particle size and PDI(DLS) are reported. PDI(DLS) is used to describe the width of the particle size distribution and is calculated from the cumulant analysis of the intensity autocorrelation function measured by DLS, and is related to the standard deviation of a hypothetical Gaussian distribution (i.e., PDI(DLS) = σ² / ZD², where σ is the standard deviation and ZD is the Z-mean particle size).
[0124]
[0186] The appearance of the nanostructures was observed using an HT-7700 transmission electron microscope at an acceleration voltage of 80 kV, a spot size of 1, and room temperature. A typical TEM grid preparation was performed as follows: The sample was diluted at room temperature with Milli-Q water to approximately 0.02–0.05 wt% of the total sample volume. A copper TEM grid pre-coated with a form bar was immersed in this solution, excess droplets were absorbed, and then it was air-dried before TEM imaging.
[0125]
[0187] Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy (ATR-FTIR): ATR-FTIR spectra were acquired using a Nicolet Nexus 870 FT-IR with a single-reflection horizontal diamond ATR accessory. The spectra were obtained at 4000 cm⁻¹. -1 ~500cm -1 The area was scanned 32 times, with a resolution of 4 cm. -1 , OPD speed 0.6289cm / s -1 The data was recorded by directly pressing a solid onto the diamond internal reflective element of the ATR, and no further sample preparation was performed.
[0126]
[0188] Synthesis of 1,1'-(azandiylbis(propane-3,1-diyl))diguanidine hydrobromide TIFF2026517422000038.tif30170
[0190] Isobutyl bromide (34.26g, 2.50x10 -1 (mol) to thiourea (19.03g, 2.50x 10-1 It was added to 100 mL of an EtOH suspension (1.00 x 10⁻⁶ mol), and the mixture was refluxed at 80°C for 5 hours. Over time, the heterogeneous mixture became homogenized, and the solution was then cooled to room temperature. Bis(3-aminopropyl)amine (13.12 g, 1.00 x 10⁻⁶ mol) -1(mol) was added, and the solution was stirred at room temperature for 15 hours. Initially, the solution was heterogeneous, but after stirring for 45 minutes it became homogeneous, and then after stirring for another 60 minutes a precipitate was observed. 300 mL of diethyl ether was added to this stirred solution, and the suspension was stirred for a further 30 minutes, after which it was filtered and washed with diethyl ether (100 mL x 2). The residue was dried under high pressure to obtain 1,1'-(azandiylbis(propane-3,1-diyl))diguanidine hydrobromide as a white solid in nearly quantitative yield. 1 ¹H NMR (400 MHz, D2O): δ 3.25 (t, J = 6.9 Hz, 4H), 2.68 (t, J = 7.5 Hz, 4H), 1.80 (p, J = 7.0 Hz, 4H); ¹³C NMR (10¹ MHz, D2O): δ 156.8, 45.5, 39.1, 27.5. See Figures 3 and 4.
[0127]
[0191] Synthesis of 2-chloro-N,N-bis(3-guanidinopropyl)acetamide hydrobromide TIFF2026517422000039.tif28170
[0192] 1,1'-(azandiylbis(propane-3,1-diyl))diguanidine hydrobromide (22.63g, 6.00x10) in 200mL of dried DMF -2 Suspend (mol) and cool to 0°C, then add triethylamine (18.22g, 1.80x10) -1 Add (mol), followed by 2-chloroacetyl chloride (20.33g, 1.80x10) -1A mol (of 10¹⁴) was added. The reaction was stirred at 0°C for about 1 hour, then warmed to room temperature. To stop the reaction of excess acyl chloride, 4.85 mL of methanol was added to the solution. The solution was stirred for a further 30 minutes, and then filtered to remove the triethylamine salt. The concentrated filtrate was then concentrated by distillation using a short-pass distillation apparatus (65°C, <1 mBar) to remove excess DMF. The viscous residue was precipitated with methanol and excess DCM (500 mL x 2). The precipitate was filtered, further washed with DCM, and dried under reduced pressure to obtain 2-chloro-N,N-bis(3-guanidinopropyl)acetamide hydrobromide as an off-white solid (yield = 57%, 15.6 g, 3.44 x 10¹⁴). -2 mol 1 H NMR (400 MHz, D2O) δ 4.28 (s, 2H), 3.44-3.35 (m, 4H), 3.20 (t, J = 6.8 Hz, 2H), 3.13 (t, J = 6.8 Hz, 2H), 1.90 (p, J = 7.0 Hz, 2H), 1.80 (p, J = 6.8 Hz, 2H); 13 ¹³C NMR (10¹ MHz, D₂O) δ 169.4, 156.8, 156.8, 45.6, 43.6, 41.3, 38.6, 38.4, 27.0, 25.7 (see Figures 5 and 6).
[0128]
[0193] Synthesis of macro CTA -- Macro CTA-A(PNIPAM44-S(C=S)SC4H9) TIFF2026517422000040.tif37170
[0194] In a clean, dry round-bottom flask, recrystallized NIPAM (106.8 mmol) and AIBN (0.238 mmol) were added as solids, followed by the addition of RAFT agent (2.38 mmol) dissolved in 25 mL of EtOH. The mixture was stirred until all components were dissolved, and the resulting solution was degassed with argon (g) for approximately 1 hour. The polymerization mixture was placed in an oil bath preheated to 60°C and polymerized for 15.5 hours. A sample was taken and shown in Figure 6. 1The conversion rate was measured by 1H NMR. Another sample was taken and purified by dialysis (MWCO 3.5kDa, against tap water, 24 hours, 5 buffer changes) for molecular weight analysis by SEC shown in Figure 7. Subsequently, this purified sample is shown in Figure 8. 1 Analysis was performed by 1H NMR.
[0129]
[0195] The molecular weight characteristics of macro CTA-A (PNIPAM44) were measured by triple detection (TD) and refractive index (RI) detection size exclusion chromatography (SEC), and the results are shown in Table 1. TIFF2026517422000041.tif35170
[0130]
[0196] Macro CTA-A:P(NIPAM) 45 ) synthesis TIFF2026517422000042.tif30170
[0197] Macro CTA-A was synthesized as follows: The concentration ratio of NIPAM / MCEBTTC RAFT agent / AIBN was 45 / 1 / 0.15, and the ratio of ethanol to NIPAM was maintained at 2 / 1 (v / w). NIPAM (20.22 g, 1.79 × 10⁻⁶) -1 Dissolve the mol) in 38.9 mL of ethanol and stir with basic alumina (150 mg) for 30 minutes to remove the inhibitor. After filtering this mixture, MCEBTTC RAFT agent (1.02 g, 4.04 x 10) was added. -3 (mol) and AIBN (98mg, 5.97x10) -4 The solution was degassed by bubbling with argon for 60 minutes after adding (mol) of the solution. Polymerization of macro CTA-A was carried out at 60°C for 15.5 hours. The reaction was stopped by exposure to air, and the solution was then used in the emulsion polymerization process. The molecular weight characteristics of macro CTA-A (PNIPAM45) were measured by TD and RI detection SEC, and are shown in Table 2. TIFF2026517422000043.tif37170
[0131]
[0198] Macro CTA-B(P(NIPAM51-co-DMAEMA29)-S(C=S)SC4H9) TIFF2026517422000044.tif38170
[0199] Recrystallized NIPAM, purified DMAEMA (by passing through a basic Al2O3 column), and AIBN initiator were added to a round-bottom flask, followed by the addition of RAFT agent dissolved in 25 mL of EtOH. The mixture was stirred until all components were dissolved, and the resulting solution was degassed with argon (g) for approximately 1 hour. The polymerization mixture was placed in an oil bath preheated to 70°C and polymerized for 15.5 hours. A sample was taken and shown in Figure 9. 1 The conversion rate was measured by 1H NMR. Another sample was taken and purified by dialysis (MWCO 3.5kDa, against tap water, 24 hours, 5 buffer changes) for molecular weight analysis by SEC shown in Figure 10. Subsequently, this purified sample is shown in Figure 11. 1 Analysis was performed by 1H NMR.
[0132]
[0200] Macro CTA-B(P(NIPAM 44 -co-DMAEMA 30 The molecular weight characteristics of )-RAFT) were measured using triple detection (TD) and refractive index (RI) detection SEC, and are shown in Table 3. TIFF2026517422000045.tif35170
[0133]
[0201] Macro CTA-B:P(NIPAM) 50 -co-DMAEMA 30 ) synthesis TIFF2026517422000046.tif25170
[0202] Macro CTA-B was synthesized as follows: The concentration ratio of NIPAM / DMAEMA / MCEBTTC RAFT agent / AIBN was 50 / 30 / 1 / 0.15, and the ratio of ethanol to NIPAM was maintained at 2 / 1 (v / w). NIPAM (22.49 g, 1.99 × 10⁻⁶) -1 (mol) and DMAEMA (18.77g, 1.19 x 10) -1 The mol) was dissolved in 52.5 mL of ethanol and stirred with basic alumina (150 mg) for 30 minutes to remove the inhibitor. After filtering this mixture, the MCEBTTC RAFT agent (1.02 g, 4.03 x 10) was added. -3(mol) and AIBN (101 mg, 6.15 x 10) -4 The solution was degassed by bubbling with argon for 60 minutes after adding (mol) of the solution. Polymerization of macro CTA-B was carried out at 70°C for 15.5 hours. The reaction was stopped by exposure to air, and the product was used directly in the emulsion polymerization process. The molecular weight characteristics of macro CTA-B P (NIPAM50-co-DMAEMA30) were measured by TD and RI detection SEC, and are shown in Table 4. TIFF2026517422000047.tif45170
[0134]
[0203] Synthesis of multifunctional nanoworms (NW) A -- Synthesis of base nanoworms (nanoworms)
[0204] Referring to Figure 8, the emulsion polymerization of styrene using macro-CTA-A and macro-CTA-B was as follows: Macro-CTA-A (2.38 mmol) and macro-CTA-B (1.72 mmol), dissolved in ethanol, were transferred to a 1 L round-bottom flask (i.e., directly from the polymerization without purification). A mixture of ice water (506 mL) was added, and the polymerization mixture was placed in an ice water bath while vigorously stirring. A surfactant (SDS) (4.28 mmol) was added in powder form, and the mixture was stirred for a further 30 minutes while degassing with argon. Then, AIBN (0.614 mmol) dissolved in styrene (229.4 mmol) was added to the mixture, and an emulsion was formed while degassing with argon (g) for a further 1 hour. The polymerization mixture was placed in an oil bath preheated to 70°C and polymerized for 7 hours. A sample was taken. 1 The conversion rate was measured by 1H NMR (CDCl3), and the styrene conversion rate reached 90%. To remove unpolymerized styrene, the remaining polymerization system was exposed to air for about 4 hours. The resulting latex was then subjected to a rotary evaporator to remove ethanol (60 mBar, 60°C bath, 4 hours) and stored in a Schott bottle. The polymer weight percentage was determined to be 9.7% from three lyophilized samples. Aliquots of the latex were dialyzed (MWCO 3.5 kDa, versus tap water, 5 buffer changes over 24 hours) and lyophilized for NMR and SEC analysis. A sample of the crude emulsion polymerization mixture is shown in Figure 9. 1Analysis was performed by 1H NMR. Another sample was taken and purified by dialysis (MWCO 3.5kDa, against tap water, 24 hours, 5 buffer changes) for molecular weight analysis by SEC as shown in Figure 10, and then freeze-dried (RI detector, DMA as eluent). Subsequently, this purified sample is shown in Figure 11. 1 Analysis was performed by 1H NMR.
[0135]
[0205] The molecular weight characteristics of the block copolymers formed after emulsion polymerization were measured using triple detection (TD) and refractive index (RI) detection SEC, and are shown in Table 5. TIFF2026517422000048.tif52170
[0136]
[0206] The obtained latex was analyzed by TEM. After cooling the sample from 70°C to room temperature, it was diluted with H2O (4 μL / mL v / v latex:H2O) (Figure 12A). As shown in Figure 12B, toluene was added (20 μL of toluene was added to 1 mL of latex), and another sample was taken at 70°C and cooled to room temperature while shaking. Then, as shown in Figure 12C, the toluene was removed by rotovap (<22°C, 30 mBar, 4 hours). Subsequently, the NW latex was diluted in aqueous solution to 8.2% by weight of nanoworms and stored in a refrigerator. The pH of the nanoworm latex was 9.5.
[0137]
[0207] Synthesis of multifunctional nanoworms (NW) B -- Synthesis of base nanoworms (nanoworms)
[0208] Referring to Figure 17, the emulsion polymerization of styrene using macro-CTA-A and macro-CTA-B was as follows: Macro-CTA-A solution (18.56 g) (MacroCTA-A+EtOH) 7.52g (MacroCTA-A) , 1.45x10 -3 mol (MacroCTA-A) ), Macro CTA-B (23.53g) (MacroCTA-B+EtOH) 11.25g (MacroCTA-B) , 1.13x10 -3 mol (MacroCTA-B) ) and SDS (0.77g, 2.66x10 -3AIBN (64 mg, 3.87 x 10) was dissolved in 300 mL of cold water at 4°C for 24 hours. This solution was degassed by bubbling with argon for 45 minutes. Subsequently, AIBN (64 mg, 3.87 x 10) was added. -4 (mol) and styrene (15.17g, 1.44x10) -1 A solution of (mol) was injected into the polymerization mixture. This mixture was further degassed by bubbling with argon for 15 minutes, and then polymerized at 70°C for 5 hours. This reaction was stopped by exposing it to ambient air at 70°C for 4 hours, and then the polymer latex was cooled to 25°C and left for 24 hours to obtain approximately 11.5% by weight of polymer latex. This polymer is shown in Figure 18 and Table 6. 1 Analysis was performed by 1H NMR. TIFF2026517422000049.tif66170
[0138]
[0209] The obtained latex was analyzed by TEM. After cooling the sample from 70°C to room temperature (e.g., approximately 20°C) o Approximately 25 minutes from C o C) Diluted with H2O (4 μL / mL v / v latex:H2O) (Figure 19).
[0139]
[0210] Morphology of nanoworms at 50°C
[0211] Nanoworms were subjected to a functionalization test in water at 50°C. First, the stability of the nanoworms at 50°C was tested. 1 mL of nanoworm latex (8.2 wt%) was heated to 50°C, stirred for 1 hour, and then cooled to room temperature. The resulting latex was examined by TEM (Figure 20A). Subsequently, the latex (1 mL) was diluted to 4.1 wt% with water (1 mL) and heated at 50°C for 1 hour. The resulting latex was then cooled to room temperature, and a TEM image was taken as shown in Figure 20B. The morphology of the nanoworms in the two weight fractions of the nanoworm dispersion was maintained and stable under these experimental conditions.
[0140]
[0212] Nanostructure synthesis from diguanidine-functionalized nanoworms
[0213] Here, the synthetic pathway to the nanostructure is described with reference to Figure 21. Stepwise quaternization and the "click" method were carried out at room temperature as follows: (i) propargyl bromide, (ii) 1-iodooctane, (iii) coumarin azide for CuAAC, and (iv) polysaccharide azide for CuAAC. For CuAAC, a combination of CuSO4 and sodium ascorbate was used. Subsequently, the final step was quaternization with digua-Cl by heating the functional nanoworm to 50°C. To test reproducibility, two replication experiments were performed and labeled as reaction A and reaction B.
[0141]
[0214] Quaternization of nanoworms with diguanidine-chloroacetamide (60°C)
[0215] Referring to Figure 22, latex containing spherical particles (9.7 wt%, 7.84 g latex, 760 mg polymer, 1.0 mmol DMAEMA units) was obtained from each of reactions A and B and weighed in a round-bottom flask equipped with a stirring bar. To this mixture, a solution of NaI (56 mg, 0.37 mmol) in H2O (2 mL) and solid digua-Cl (178 mg, 0.37 mmol) were added. After taking a sample (200 μL) for NMR analysis shown in Figure 23, the reaction flask was placed in an oil bath preheated to 60°C before the reaction to track the changes in digua-Cl. The loss of α-protons from digua-Cl and the loss of CH3 protons from the DMAEMA side group (b) after 14 hours are shown. After stirring the reaction for 14 hours, an aliquot (200 μL) was taken for NMR analysis.
[0142]
[0216] Samples were taken from both reactions (e.g., reaction A and reaction B), dialyzed (MWCO 3.5 kDa, against tap water, 24 hours, 5 buffer changes), and lyophilized for NMR analysis as shown in Figure 24.
[0143]
[0217] Table 7 shows the binding efficiency of reactions A and B of diguanidine-chloroacetamide to spherical particles. TIFF2026517422000050.tif42170
[0144]
[0218] Quaternation of nanoworms with diguanidine chloroacetamide at 50°C before functionalization with propargyl bromide, 1-iodooctane, polysaccharide azide, and coumarin azide.
[0219] Referring to Figure 25, nanoworm latex (8.2 wt%, 3.2 g, 0.3 mmol DMAEMA units) and H2O (3 mL) were added to obtain an aqueous latex solution containing 4.1 wt% nanoworm latex at room temperature. Diguanidine chloroacetamide (digua-Cl solid, 52.1 mg, 0.115 mmol) and NaI (17.3 mg, 0.115 mmol) were added to the latex, and the mixture was placed in an oil bath at 50°C and stirred at 100 rpm for 14 hours (i.e., overnight). Based on the NMR analysis of the crude mixture shown in Figure 26, the conversion rate of digua-Cl when nanoworms were quaternized with diguanidine was 54% (after 14 hours), and this increased to 92% after 36 hours. The morphology after quaternization did not show any change to the nanoworm structure, as shown in Figures 27A and 27B.
[0145]
[0220] Quaternization of nanoworms by propargyl bromide
[0221] Referring to Figure 28, a toluene solution of propargyl bromide (80% by weight, 17.4 mg in 500 μL EtOH) was added to each mixture of reaction A and reaction B described above, and the reaction was stirred at room temperature at a stirring speed of less than 50 rpm. 1 As shown in Figure 29, the propargyl bromide was completely consumed to below the detection limit of the NMR spectrum within 2 hours by 1H NMR. The reaction was allowed to stand for a further 4 hours before proceeding to the next step.
[0146]
[0222] Samples were collected, purified by dialysis (MWCO 3.5kDa, against tap water, 24 hours, 5 buffer changes), and freeze-dried to measure binding efficiency. Although not constrained by any particular theory, and likely due to the protonation of DMAEMA units after quaternization, the binding efficiency based on the integration of DMAEMA shows a value higher than 100%, as shown in Figure 30.
[0147]
[0223] Table 8 shows the binding efficiency of propargyl bromide to spherical particles in reactions A and B. TIFF2026517422000051.tif42170
[0148]
[0224] Quaternization of nanoworms by 1-iodooctane
[0225] Referring to Figure 31, a solution of 1-iodooctane in ethanol (30.4 mg in 0.395 mL) was added to the latex obtained from each of reaction A and reaction B, and the mixture was kept at room temperature overnight, for example, for about 8 to 16 hours. The sample was taken and purified by dialysis (MWCO 3.5 kDa, against tap water, 24 hours, 5 buffer changes), as shown in Figure 32. 1 Analysis was performed by 1H NMR. The binding efficiency was over 100%. Table 9 shows the binding efficiency of 1-iodooctane to spherical particles for reactions A and B. TIFF2026517422000052.tif42170
[0149]
[0226] CuAAC coupling with polysaccharide azides and coumarin azides.
[0227] Referring to Figure 33, half of each latex obtained was used to carry out the CuAAC reaction. Sodium ascorbate (38 mg), polysaccharide azide solid (Mn3200, 107 mg), and coumarin azide (5.1 mg in 250 μL EtOH) were added to each latex. The resulting mixture was degassed by argon bubbling for 30 minutes. Then, degassed copper sulfate pentahydrate aqueous solution (48 mg, 0.19 mmol, 2.85 mL of 100 mg / 6 mL solution) was added by syringe. The reaction was maintained under argon for 24 hours. After the CuAAC click reaction, as shown in Figure 34, the polysaccharide signal was 1Almost no protons were observed by 1H NMR. The sample before CuAAC was taken from the latex obtained by adding only polysaccharide-N3 in the final step. The sample after CuAAC was treated with neutral Al2O3 (stirred together with a portion of the latex, and then the supernatant was used). When polysaccharide-N3 was added to the latex without the addition of a copper catalyst, protons derived from the polysaccharide were detected. The proton signal was reduced because the mobility of the polysaccharide bound to the nanoworm was restricted.
[0150]
[0228] After the reaction, the mixture was exposed to air, and an aqueous solution of trisodium EDTA salt was added to each reaction (0.5 M, 381 μL, 0.19 mmol, 1 equivalent relative to the CuSO4·5H2O added above). Since the pH was found to be less than 7.0 due to oxidized sodium ascorbate, solid K2CO3 (74 mg) was added to adjust the basicity to pH 9-10 in preparation for the next reaction step.
[0151]
[0229] CuAAC click reaction between poly(galactose)-azide and 3-azide-7-hydroxycoumarin
[0230] As shown in Figure 35, from the latex mixture (i.e., after quaternization), a 1 mL fraction (containing 0.015 mmol of propargyl groups) was taken, to which a solution of PGal-N3 (Mn=3200, 27.2 mg, 0.0085 mmol) and 7HC-N3 (1.3 mg) in EtOH (100 μL) dissolved in H2O (1 mL) was added and stirred to mix thoroughly. A sample (200 μL) of this crude mixture was immediately taken for NMR analysis. Solid ascorbic acid (17 mg, 0.1 mmol, 6 equivalents relative to the above propargyl bromide) was added, and the mixture was degassed under argon for about 15 minutes. In a separate round-bottom flask, CuSO4 (24.2 mg, 6 equivalents relative to the propargyl groups) was dissolved in H2O (2 mL), and the mixture was degassed for about 15 minutes. This CuSO4 was transferred to nanoworm latex using a syringe. Prior to the TEM analysis shown in Figures 36A and 36B, the reaction was gently stirred at 50 rpm under argon for 24 hours. These samples were then dialyzed (in water containing MWCO 10 kDa, 0.2 wt% EDTA-Na3 salt, with 5 buffer changes over 48 hours, followed by 4 hours in milli-Q water).
[0152]
[0231] 1 ¹H NMR was performed to measure the changes in polysaccharides before and after the CuAAC "click" reaction. After coupling, as shown in Figures 37 and 38, the polysaccharides became much harder to observe in the NMR of the crude mixture, and were no longer observable after purification.
[0153]
[0232] When D2O was added, polysaccharide protons could be observed, while signals from polystyrene and other molecules were suppressed, allowing triazole H-5 to be observed (Figures 39 and 40).
[0154]
[0233] Quaternation of nanoworms with diguanidine chloroacetamide at 50°C after functionalization with propargyl bromide, 1-iodooctane, polysaccharide azide, and coumarin azide.
[0234] Referring to Figure 41, digua-Cl solid (87 mg, 0.19 mmol) and NaI (28.8 mg, 0.19 mmol) were added to the latex. The reaction was then heated at 50°C for 36 hours. TEM showed that the nanoworm structure was maintained after the reaction (shown in Figure 42). Small preparative samples were taken for NMR analysis to determine the binding efficiency and purified by dialysis (MWCO 10 kDa, tap water, 24 hours, 5 buffer changes) (shown in Figure 43). As shown in Table 10, the binding efficiencies in the replication experiments (e.g., reaction A and reaction B) were 17% and 39%, respectively. TIFF2026517422000053.tif42170
[0155]
[0235] Quaternization of nanoworms at high temperatures using polysaccharide pseudohalides, diguanidine-chloroacetamide, and iodooctane components.
[0236] Referring to Figure 44, a method for binding polysaccharides via quaternization rather than CuAAC is described. By not using ascorbate and EDTA, the binding efficiency of diguanidine chloroacetamide can be improved.
[0156]
[0237] Preparation of quaternary worms from base spherical particles (synthesis of nanostructure A A-E, small scale, 10 mL)
[0238] Referring to Figures 45A-F, nanostructure A was synthesized using multiple synthesis procedures, e.g., synthesis A, synthesis B, synthesis C, synthesis D, synthesis E, and synthesis F. Polymer latex (11.5 wt%, 11.4 g latex, 1.3 g polymer, 1.30 mmol DMAEMA units) was placed in a water bath controlled at 60°C. To this mixture, iodooctane (40.6 mg, 1.69 x 10¹³) dissolved in 1.4 mL of EtOH was added. -4 Add (mol) and stir the reaction for 24 hours. Next, add propargyl bromide (23.2 mg, 1.56 x 10) dissolved in 1.4 mL of EtOH. -4 The reaction was stirred for 24 hours after adding 72 mg of NaI (4.81 x 10) dissolved in 0.36 mL of water. -4Diguanidine-Cl(140.3 mg, 3.09 x 10) dissolved in 0.70 mL of water (mol) -4 (mol) was added, and the reaction was stirred for 24 hours. After the reaction was complete, the polymer latex was cooled to 25°C. Table 11 shows the visualization of colloidal stability during the quaternization reaction. TIFF2026517422000054.tif62170
[0157]
[0239] Fourth-class efficiency 1 The results were measured by 1H NMR and are shown in Table 12 below. TIFF2026517422000055.tif62170
[0158]
[0240] Preparation of quaternary worms from base spherical particles (synthesis of nanostructure A A-E, small scale, 10 mL)
[0241] Referring to Figure 46, polymer latex (11.5% by weight, 11.4 g latex, 1.3 g polymer, 1.30 mmol DMAEMA units) was placed in a water bath controlled at 25°C. To this mixture, iodooctane (40.6 mg, 1.69 x 10¹³) dissolved in 1.4 mL of EtOH was added. -4 Add (mol) and stir the reaction for 24 hours. Next, add propargyl bromide (23.2 mg, 1.56 x 10) dissolved in 1.4 mL of EtOH. -4 The reaction was stirred for 24 hours after adding 72 mg of NaI (4.81 x 10) dissolved in 0.36 mL of water. -4 Diguanidine-Cl(140.3 mg, 3.09 x 10) dissolved in 0.70 mL of water (mol) -4 (mol) was added and the reaction was stirred for 5 minutes. The reaction mixture was then heated to 60°C and stirred for 24 hours. After the reaction was complete, the polymer latex was cooled to 25°C. An increase in the viscosity of the polymer latex was observed after reaching room temperature. TEM characterization showed that the quaternized spherical particles were converted into nanoworms even without the addition of toluene as a plasticizer. Table 12 shows the visualization of colloidal stability during the quaternization reaction. TIFF2026517422000056.tif33170
[0159]
[0242] Fourth-class efficiency 1 The results were measured by 1H NMR and are shown in Table 13 below. TIFF2026517422000057.tif33170
[0160]
[0243] TEM micrographs of the quaternary grading without the addition of plasticizers are shown in Figures 47A-47C.
[0161]
[0244] Preparation of quaternary nanoworms using CuAAC
[0245] Referring to Figure 48, polymer latex (8% by weight, 11.4 g latex, 1.3 g polymer, and 0.16 mmol propargyl group) contains P (galactose) - -N3 (153.3 mg, 6.13 x 10) -5 (mol) was added and degassed by bubbling with argon for 20 minutes. 3-azido-7-hydroxycoumarin (13.9 mg, 6.83 x 10) -5 The ascorbic acid (mol) was dissolved in 4.23 mL of EtOH, degassed by bubbling with argon for 20 minutes, and then injected into the reaction mixture. -4 The mol of CuSO4 was dissolved in 1 mL of water, degassed by bubbling with argon for 20 minutes, and then injected into the reaction mixture. CuSO4(149.3 mg, 9.35 x 10) -4 The solution (mol) was dissolved in 1 mL of water, degassed by bubbling with argon for 20 minutes, and then injected into the reaction mixture. The CuAAC reaction was stirred overnight under argon, and then purified by dialysis against water (MWCO 10 kDa) for 12 hours. TEM images are shown in Figures 49A-49C.
[0162]
[0246] Conversion from spherical particles to nanoworms
[0247] Referring to Figure 50, for each reaction mixture, a sample (200 μL × 4 = 800 μL) was taken for analysis at each step, leaving 7 mL of latex remaining from the original latex (7.8 mL). Toluene (140 μL, 20 μL / mL toluene: latex v / v) was added to each latex mixture at 60°C. The resulting mixtures were cooled to room temperature and allowed to stand for 24 hours. Subsequently, toluene was removed using a rotary evaporator (20°C water bath, 30 mBar, 2 hours). Since water was lost during toluene removal, the mixture was diluted back to 7 mL. After cooling, TEM images were acquired and shown in Figures 51A and 51B.
[0163]
[0248] Fluorescence test
[0249] Fluorescence tests were performed using a fluorescence spectrophotometer to measure fluorescence emission at different concentrations of nanoworms. A control was used for comparison.
[0164]
[0250] The freeze-dried samples were weighed and then suspended in H2O at room temperature to obtain a 1.5% by weight suspension. This mixture was placed in an ice bath for 1 hour and shaken at room temperature for 1 hour. Subsequently, this suspension was diluted to different concentrations. Prior to the fluorescence tests shown in Table 14, samples of different concentrations were added to 96-well plates (200 μL each). TIFF2026517422000058.tif98170
[0165]
[0251] Preparation of nanostructure A (large scale, reproducibility)
[0252] Synthesis of macro CTA-A:P(NIPAM45) TIFF2026517422000059.tif29170
[0253] Macro CTA-A was synthesized as follows: The concentration ratio of NIPAM / MCEBTTC RAFT agent / AIBN was 45 / 1 / 0.15, and the ratio of ethanol to NIPAM was maintained at 2 / 1 (v / w). NIPAM (7.20 g, 6.37 × 10⁻⁶) -2The solution (mol) was dissolved in 14.4 mL of ethanol and stirred with basic alumina (50 mg) for 30 minutes to remove the inhibitor. After filtering this mixture, the MCEBTTC RAFT agent (357.5 mg, 1.42 x 10) was extracted. -3 (mol) and AIBN (35 mg, 2.13 x 10) -4 The solution was degassed by bubbling with argon for 40 minutes. Polymerization of macro CTA-A was carried out at 60°C for 15.5 hours. The reaction was stopped by exposure to air, and the product was used directly in the emulsion polymerization process. Molecular weight analysis was performed, and the results are shown in Table 15. TIFF2026517422000060.tif29170
[0166]
[0254] Macro CTA-B: P(NIPAM) 50 -co-DMAEMA 30 ) synthesis
[0255] Macro CTA-B was synthesized as follows: The concentration ratio of NIPAM / DMAEMA / MCEBTTC RAFT agent / AIBN was 50 / 30 / 1 / 0.15, and the ratio of ethanol to NIPAM was maintained at 2 / 1 (v / w). NIPAM (6.32g, 5.58 × 10⁻³) -2 (mol) and DMAEMA (5.27g, 3.35 x 10) -2 The mol) was dissolved in 12.6 mL of ethanol and stirred with basic alumina (50 mg) for 30 minutes to remove the inhibitor. After filtering this mixture, the MCEBTTC RAFT agent (281.4 mg, 1.11 x 10) was extracted. -3 (mol) and AIBN (27.8 mg, 1.69 x 10) -4 The solution was degassed by bubbling with argon for 40 minutes. Polymerization of macro CTA-B was carried out at 70°C for 15.5 hours. The reaction was stopped by exposure to air and the product was used directly in the emulsion polymerization process. Molecular weight analysis was performed, and the results are shown in Table 16. TIFF2026517422000061.tif39170
[0167]
[0256] Preparation of spherical base particles by emulsion polymerization (300 mL scale)
[0257] Referring to Figure 52, the emulsion polymerization of styrene using macro-CTA-A and macro-CTA-B was as follows: Macro-CTA-A solution (18.96g) (MacroCTA-A+EtOH) 7.51g (MacroCTA-A) , 1.42x10 -3 mol (MacroCTA-A) ), Macro CTA-B (21.87g) (MacroCTA-B+EtOH) 11.70g (MacroCTA-B) , 1.12x10 -3 mol (MacroCTA-B) ) and SDS (0.77g, 2.66x10 -3 AIBN (63 mg, 3.80 x 10) was dissolved in 300 mL of cold water at 4°C for 24 hours. This solution was degassed by bubbling with argon for 45 minutes. Then, AIBN (63 mg, 3.80 x 10) was added. -4 (mol) and styrene (14.80g, 1.42x10) -1 A solution of (mol) was injected into the polymerization mixture. This mixture was further degassed by bubbling with argon for 15 minutes, and then polymerized at 70°C for 5 hours. The reaction was stopped by exposing it to 70°C air for 4 hours, and then the polymer latex was cooled to 25°C and left for 24 hours. Molecular weight analysis and SEC trace analysis were performed, and the results are shown in Table 17 and Figure 53. TIFF2026517422000062.tif39170
[0168]
[0258] Preparation of nanostructured A quaternary worms from base spherical particles (large scale, 300 mL)
[0259] Here, referring to Figure 54, polymer latex (300 mL, 3.35 × 10 DMAEMA units) -2 The mixture was placed in a water bath controlled at 25°C. To this mixture, iodooctane (1.0453 g, 4.35 x 10) dissolved in 27.5 mL of EtOH was added. -3 Next, add (0.5975 g, 4.02 x 10) of propargyl bromide dissolved in 27.5 mL of EtOH and stir the reaction for 24 hours. -3 The reaction was stirred for 24 hours after adding NaI(1.8571g, 1.24x10) dissolved in 9.3 mL of water. -2Diguanidine-Cl(3.6152g, 7.97x10) dissolved in 18.1 mL of water (mol) -3 (mol) was added and the reaction was stirred for 5 minutes. The reaction mixture was then heated to 60°C and stirred for 24 hours. After the reaction was complete, the polymer latex was cooled to 25°C. An increase in the viscosity of the polymer latex was observed after reaching room temperature. TEM characterization showed that the quaternized spherical particles were converted into nanoworms even without the addition of toluene as a plasticizer. Table 18 shows the visualization of colloidal stability during the quaternization reaction. TIFF2026517422000063.tif33170
[0169]
[0260] Fourth-class efficiency 1 The results were measured by 1H NMR and are shown in Table 19 below. TIFF2026517422000064.tif33170
[0170]
[0261] TEM micrographs of the quaternary grading without the addition of plasticizers are shown in Figures 55A-55C.
[0171]
[0262] Virucidal activity
[0263] Referring to Figures 56A-56D, the samples on the trays prepared for virus testing were either uncoated or coated with nanostructure A sprayed five times. The trays were analyzed under natural light and ultraviolet light.
[0172]
[0264] To measure the antiviral activity, several samples were analyzed as shown in Table 20.
[0265] TIFF2026517422000065.tif127170
[0173]
[0266] Viral activity of nanostructure A against SARS-CoV-2
[0267] The inoculum was prepared by adding an equal volume of virus stock to filter-sterilized Sørensen pH buffer at pH 6.5. 50 μL of the inoculum was dropped onto the provided tray (2 × 2 cm). The sample was incubated for 30 minutes (room temperature, with the plate covered in a BSC Class II hood). After 30 minutes of incubation (step 2), 0.5 mL of infection medium (MEM + antibiotic) was added to the sample, and unbound virions / viral debris were removed from the coating surface by vigorous pipetting. The surface was not damaged by pipetting. The medium was collected to quantify infectious virus titers by TCID50 (P13 and untreated samples only) and qRT-PCR (all samples). TCID50 was performed immediately after sample collection. For RNA extraction, the sample was immediately placed in lysis buffer, and RNA was extracted. The RNA was stored at -80°C until processed by qRT-PCR.
[0174]
[0268] 50% Tissue Culture Infectious Dose Assay (TCID) 50 The assay was performed according to the following protocol. 24 hours prior to the assay, approximately 95% of Vero cells were seeded onto plates to form a monolayer. After confirming the quality / density of the monolayer membrane, the plates were washed with infection medium (to remove cell debris) and transferred to the PC3 laboratory. Samples were prepared, serially diluted, and then inoculated into each well with a known amount of MEM infection medium (containing pen / strep, glutamine, and HEPES, but without FBS) + TPCK trypsin (1 ug / mL). The plates were returned to the incubator (37°C, 5% CO2), and cytopathic effects (CPE) on cells were examined microscopically on post-infection day 5 (Omicron takes 48 hours longer to induce CPE in Vero cells than other VOC isolates). To confirm the presence or absence of virus-induced CPE, the samples were subjected to a second passage in Vero cells. Subsequently, the TCID50 / mL of infectious virus present in the original samples was measured.
[0175]
[0269] Quantitative real-time polymerase chain reaction (qRT-PCR) was performed on omicron, delta, and alpha variants of SARS-CoV-2. RNA was extracted from samples using the commercially available QiaAmp Virus RNA Mini Extraction Kit from Qiagen, following the manufacturer's instructions. Samples were stored at -80°C until ready for processing. Reaction solutions for detecting the SARS-CoV-2 envelope (E) gene were prepared, containing known amounts of standards, thawed samples, and controls, and processed using an RT-PCR instrument under appropriate thermal cycling conditions. The amount of genomic RNA present in each sample was determined using CT (optical density) values and interpolated from the created standard curve. Figures 57-60 show the results of significant degradation of the SARS-CoV-2 omicron, delta, and alpha genomes by nanostructure A.
[0176]
[0270] Viral activity of nanostructure A against influenza type A
[0271] The inoculum was prepared by diluting a high-concentration egg stock of PR8 in filter-sterilized Sørensen pH buffer at pH 6.5. 50 μL of the inoculum was dropped onto a provided tray (2 × 2 cm). The sample was incubated for 30 minutes (room temperature, with the plate covered in a BSC class II hood). After 30 minutes of incubation (step 2), 0.5 mL of infection medium (RPMI + antibiotic) was added to the sample and vigorously pipettered to remove unbound virions / viral debris from the coating surface. The surface was not damaged by pipetting. The medium was collected for quantification of infectious virus titers by TCID50 and qRT-PCR. TCID50 was performed immediately after sample collection. For RNA extraction, the sample was immediately placed in lysis buffer, and RNA was extracted. The RNA was stored at -80°C until processed by qRT-PCR.
[0177]
[0272] As shown in Figures 61 and 62 and Table 21, a significant reduction was observed in all samples exposed to nanostructure A compared to the uncoated control.
[0273] TIFF2026517422000066.tif79170
[0178] Additional aspects
[0274] This disclosure provides, but is not limited to, the following embodiments. Each embodiment may be deemed to optionally include any alternative embodiment.
[0179]
[0275] Clause 1. A nanostructure comprising a compound or a salt thereof, wherein the compound is Multiple N-isopropylacrylamide units; the part represented by the following formula: TIFF2026517422000067.tif19170 (in the formula, R 1 is C1-C 20 It is alkyl; The part represented by the following equation: TIFF2026517422000068.tif28170 (in the formula, R 2 , R 3 and R 4 Each is independently hydrogen or C1-C 20 It is alkyl; Multiple parts represented by the following equation: TIFF2026517422000069.tif20170 (in the formula, R 5 is C1-C 10 It is aryl; and Multiple parts represented by the following equation: TIFF2026517422000070.tif61170(In the formula, Q is O or N, R ** is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. 6 , R 7 and R 8 R is a C1-C6 alkyl group or hydrogen, 9 If present, C1-C 16Alkyl, C1-C6 alkylene, azole, guanidine, guanidine oligomer (e.g., diguanidine), polysaccharide, chromophore, and one or more combinations thereof, with at least one R 9 (There exists a digunidine.) Nanostructures, including
[0180]
[0276] Clause 2. A nanostructure as described in Clause 1, wherein the nanostructure is of formula: TIFF2026517422000071.tif56170(wherein n, m and p are each an integer between 1 and 100 independently, q is 0 or 1, and R of equation (I) 2 , R 3 and R 4 Each of them is independently hydrogen or C1-C 20 It is alkyl, and R of formula (I) 5 is C4-C 10 It is an aryl and R in equation (I). 6 , R 7 , and R 8 R is a C1-C6 alkyl or hydrogen atom, and is part of formula (I). 9 If present, C1-C 16 Alkyl, C1-C6 alkylene, azole, guanidine, guanidine oligomer (e.g., diguanidine), polysaccharide, chromophore, and one or more combinations thereof, R 10 and R 11 Each of them is independently hydrogen or C1-C 20 A nanostructure represented by (alkyl).
[0181]
[0277] Clause 3. The nanostructure described in Clause 2, wherein n is between 20 and 50.
[0182]
[0278] Clause 4. The nanostructure described in Clause 3, wherein n is 30.
[0183]
[0279] Clause 5. A nanostructure as described in any of Clauses 2 to 4, wherein n is between 30 and 60.
[0184]
[0280] Clause 6. The nanostructure described in Clause 5, wherein m is 45.
[0185]
[0281] Clause 7. A nanostructure as described in any of Clauses 2 to 6, wherein n is between 45 and 55.
[0186]
[0282] Clause 8. The nanostructure described in Clause 7, wherein p is 50.
[0187]
[0283] Clause 9. The nanostructure described in Clause 7, wherein p is 52.
[0188]
[0284] Article 10. R 1 A nanostructure according to any of clauses 2 to 9, wherein is butyl.
[0189]
[0285] Article 11. R 2 A nanostructure according to any of clauses 2 to 10, wherein is methyl.
[0190]
[0286] Article 12. R 3 A nanostructure according to any of clauses 2 to 11, wherein is a C1-C20 alkyl group.
[0191]
[0287] Article 13. R 3 The nanostructure described in Clause 12, wherein the molecule is methyl.
[0192]
[0288] Article 14. R 3 A nanostructure as described in Clause 12, wherein hydrogen is present.
[0193]
[0289] Article 15. R 4 A nanostructure according to any of clauses 2 to 14, wherein is a C1-C20 alkyl group.
[0194]
[0290] Article 16. R 4 The nanostructure described in Clause 15, wherein the molecule is methyl.
[0195]
[0291] Article 17. R 4 A nanostructure as described in Clause 15, wherein hydrogen is present.
[0196]
[0292] Article 18. R 5 A nanostructure as described in any of clauses 2 to 17, wherein is phenyl.
[0197]
[0293] Article 19. R 6 However, the nanostructure is a C1-C6 alkyl group, as described in any of clauses 2 to 18.
[0198]
[0294] Article 20. R 6 The nanostructure described in Clause 19, wherein the molecule is methyl.
[0199]
[0295] Article 21. R 7 However, the nanostructure is a C1-C6 alkyl group, as described in any of clauses 2 to 20.
[0200]
[0296] Article 22. R 7 The nanostructure described in Clause 21, wherein is methyl.
[0201]
[0297] Article 23. R 8 However, the nanostructure is a C1-C6 alkyl group, as described in any of clauses 2 to 22.
[0202]
[0298] Article 24. R 8 The nanostructure described in Clause 23, wherein the molecule is methyl.
[0203]
[0299] Article 25. R 9 However, C1-C 16 A nanostructure that is alkyl, as described in any of clauses 2 to 24.
[0204]
[0300] Article 26. R 9 The nanostructure described in Clause 25, wherein is a C7 alkyl group.
[0205]
[0301] Article 27. R 9 However, the nanostructure is a C1-C6 alkylene, as described in any of clauses 2 to 26.
[0206]
[0302] Article 28. R 9 The nanostructure described in Clause 26, wherein the C3 alkylene is present.
[0207]
[0303] Article 29. R 9 A nanostructure according to any one of the clauses 2 to 28, wherein is an azole.
[0208]
[0304] Article 30. R 9 The nanostructure described in Clause 29 is a combination of azole and polygalactose.
[0209]
[0305] Article 31. R 9 The nanostructure described in Clause 29 is a combination of azole and coumarin.
[0210]
[0306] Article 32. The following formula: TIFF2026517422000072.tif55170(wherein r, s, t, u, m and p are integers from 1 to 100, R 1 It is butyl, and R 2 , R 3 , R 6 , R 6’ , R 6’’ , R 6’’’ , R 7 , R 7’ , R 7’’ , R 7’’’ , R 8 , R 8’ , R 8’’ and R 8’’’ Each of them is methyl, R 4 is hydrogen, R 5 is phenyl, and R 10 It is isopropyl, and R 11 is hydrogen, R 12 It is diguanidine, and R 13 is an azole, and R 14 A nanostructure as described in any of clauses 2 to 31, represented by (where is octyl).
[0211]
[0307] Clause 33. A method for depositing nanostructures on a surface, wherein the nanostructures are Multiple N-isopropylacrylamide units; the part represented by the following formula: TIFF2026517422000073.tif19170 (in the formula, R 1 is C1-C 20 It is alkyl; The part represented by the following equation: TIFF2026517422000074.tif28170 (in the formula, R 2 , R 3 and R 4 Each is independently hydrogen or C1-C 20 It is alkyl; Multiple parts represented by the following equation: TIFF2026517422000075.tif20170 (in the formula, R 5 is C1-C 10 It is aryl; and Multiple parts represented by the following equation: TIFF2026517422000076.tif61170(In the formula, Q is O or N, R ** is C1-C 20 Alkyl, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc. 6 , R 7 and R 8 R is a C1-C6 alkyl group or hydrogen, 9 If present, C1-C 16 Alkyl, C1-C6 alkylene, azole, guanidine, guanidine oligomer (e.g., diguanidine), polysaccharide, chromophore, and one or more combinations thereof, with at least one R 9 (There exists a digunidine.) A method for depositing nanostructures, including [specific elements], onto a surface.
[0212]
[0308] Clause 34. The method according to Clause 33, wherein the deposition is carried out using an aqueous solution containing the nanostructure at a concentration of approximately 0.5% to approximately 3% by weight.
[0213]
[0309] Clause 35. The method according to Clause 33, wherein the deposition is carried out using an aqueous emulsion containing the nanostructure at a concentration of approximately 0.5% to approximately 3% by weight.
[0214]
[0310] Clause 36. The method according to any one of Clauses 33 to 35, further comprising depositing the aqueous solution on a surface and then evaporating the water from the aqueous solution.
[0215]
[0311] Clause 37. The method according to any one of Clauses 33 to 36, wherein the deposition of the above structure on a surface is carried out by coating the surface, dipping the surface, spraying the surface, taping the surface, brushing the surface, spin-coating the surface, roll-coating the surface, doctor-blade coating the surface, or a combination of one or more thereof, using the above nanostructure.
[0216]
[0312] Clause 38. The method described in any one of Clauses 33 to 37, wherein the surface is the surface of personal protective equipment.
[0217]
[0313] Clause 39. The method according to any one of Clauses 33 to 38, wherein the surface is an internal or external surface of an aircraft, ship, train, boat, terminal building, or spacecraft.
[0218]
[0314] Clause 40. The method according to any one of Clauses 33 to 39, wherein the surface is the surface of the vehicle's air filter.
[0219]
[0315] Clause 41. The method according to any one of Clauses 33 to 40, wherein the surface is a floor surface, a seat surface, a tray table surface, an overhead luggage rack surface, a ceiling surface, a door surface, or a door handle surface.
[0220]
[0316] Clause 42. The method according to any one of Clauses 33 to 41, wherein the nanostructure is a nanoworm.
[0221]
[0317] Clause 43. The method according to any one of Clauses 33 to 42, wherein the nanostructure is a nanorod.
[0222]
[0318] While aspects of this disclosure have been described, other and further aspects of this disclosure can be devised without departing from the basic scope of this disclosure, and the scope of this disclosure is defined by the following claims.
Claims
1. A nanostructure comprising a compound or a salt thereof, wherein the compound is Multiple N-isopropylacrylamide units; The part represented by the following equation: (In the formula, R １ C １ -C ２０ (It is alkyl); The part represented by the following equation: (In the formula, R ２ , R ３ and R ４ Each is independently either hydrogen or C １ -C ２０ (It is alkyl); Multiple parts represented by the following equation: (wherein, R ５ is C １ -C １０ aryl); and Multiple parts represented by the following equation: (In the formula, Q is O or N, and R ＊＊ is C １ -C ２０ It is alkyl, R ６ , R ７ and R ８ is C １ -C ６ Alkyl or hydrogen, R ９ If it exists, C １ -C １６ Alkyl, C １ -C ６ Alkylene, azole, guanidine, guanidine oligomer, polysaccharide, chromophore, and one or more combinations thereof, and at least one R ９ (It exists, and it is diguanidine.) Nanostructures, including
2. Equation (I): (In equation (I), n, m, and p are each independent integers from 1 to 100, q is 0 or 1, and R in equation (I) ２ , R ３ and R ４ Each of them is independently hydrogen or C １ -C ２０ It is alkyl, and R of formula (I) ５ is C ４ -C １０ It is aryl, and R in equation (I). ６ , R ７ , and R ８ is C １ -C ６ Alkyl or hydrogen, and R of formula (I) ９ If it exists, C １ -C １６ Alkyl, C １ -C ６ Alkylene, azole, guanidine, diguanidine, and other guanidine oligomers, polysaccharides, chromophores, and one or more combinations thereof, and R of formula (I) １０ and R １１ Each of them independently contains hydrogen or C １ -C ２０ The nanostructure according to claim 1, represented by (being alkyl).
3. The nanostructure according to claim 1 or 2, wherein n is 20 to 50.
4. A nanostructure according to any one of claims 1 to 3, wherein m is 30 to 60.
5. The nanostructure according to any one of claims 1 to 4, wherein p is 45 to 55.
6. The nanostructure according to any one of claims 1 to 5, wherein p is 52.
7. R １ The nanostructure according to any one of claims 1 to 6, wherein is butyl.
8. R ２ The nanostructure according to any one of claims 1 to 7, wherein is methyl.
9. R ３ The nanostructure according to any one of claims 1 to 8, wherein is hydrogen.
10. R ４ The nanostructure according to any one of claims 1 to 9, wherein is methyl.
11. R ５ The nanostructure according to any one of claims 1 to 10, wherein is phenyl.
12. R ６ , R ７ , and R ８ Each of them independently C １ -C ６ The nanostructure according to any one of claims 1 to 11, wherein the nanostructure is alkyl.
13. R ６ , R ７ , and R ８ The nanostructure according to claim 12, wherein each of them is independently methyl.
14. R ９ C ７ The nanostructure according to any one of claims 1 to 13, wherein the nanostructure is alkyl.
15. R ９ C ３ The nanostructure according to any one of claims 1 to 13, wherein the nanostructure is alkylene.
16. A nanostructure according to claim 2, represented by formula (II), wherein the following formula: (In the formula, r, s, t, u, m and p are integers from 1 to 100, R １ It is butyl, and R ２ , R ３ , R ６ , R ６’ , R ６’’ , R ６’’’ , R ７ , R ７’ , R ７’’ , R ７’’’ , R ８ , R ８’ , R+, and R ８’’’ Each of them is methyl, R ４ is hydrogen, R ５ is phenyl, and R １０ It is isopropyl, and R １１ is hydrogen, R １２ It is diguanidine, and R １３ is an azole, R １４ A nanostructure represented by (where is octyl).
17. A method for depositing nanostructures on a surface, comprising depositing a nanostructure containing a compound or a salt thereof on a surface, wherein the compound is Multiple N-isopropylacrylamide units; the part represented by the following formula: (In the formula, R １ C １ -C ２０ (It is alkyl); The part represented by the following equation: (wherein, R ２ , R ３ and R ４ are each independently hydrogen or C １ -C ２０ alkyl); Multiple parts represented by the following equation: (In the formula, R ５ C １ -C １０ (It is alkyl); and The following formula: A plurality of moieties represented by (where Q is O or N, and R ＊＊ is C １ -C ２０ alkyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc., and R ６ , R ７ and R ８ are C １ -C ６ alkyl or hydrogen, and R ９ is, when present, C １ -C １６ alkyl, C １ -C ６ alkylene, an oligomer of guanidine such as azole, guanidine, diguanidine, polysaccharide, chromophore, and one or a combination of more of these, and at least one R ９ is present, which is diguanidine) A method for depositing nanostructures, including [specific elements], onto a surface.
18. The method according to claim 17, wherein the deposition is carried out using an aqueous solution containing the nanostructure at a concentration of about 0.5% to about 3% by weight.
19. The method according to claim 17 or 18, further comprising depositing the aqueous solution on the surface and then evaporating the water from the aqueous solution.
20. The method according to any one of claims 1 to 19, wherein the nanostructure is a nanoworm.