Antimicrobial compounds and nanostructures

Abstract

To provide: antimicrobial compounds and nanostructures capable of further functionalization and antimicrobial activity against strong microbes; compositions including the compounds and nanostructures; and methods useful for antimicrobial and / or antiviral applications using the compounds, the nanostructures and the compositions.SOLUTION: The problem is solved by a nanorod and / or a nanoworm described in the claims.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present disclosure provides compounds and nanostructures that include one or more quaternary ammonium salts, compositions that include the compounds and nanostructures, and methods useful for treating conditions using the compounds, nanostructures and compositions. [Background technology]

[0002] Prevention of disease transmission on aircraft has traditionally focused on improving air conditioning / filtration systems. Recent studies have suggested that one way to further improve disease prevention on aircraft and spacecraft may include surface contamination treatment, for example, of aircraft surfaces. However, while some antimicrobial compounds have been produced for antibacterial applications, no compounds have been specifically demonstrated for antiviral applications. For antiviral compounds, the biotechnology and pharmaceutical industries use proteins and small molecules (e.g., molecular weight less than 1,000) that target microorganisms. However, proteins and small molecules have limited systemic half-lives, and as a result, they usually require higher than desirable dosages.

[0003] Furthermore, the hydrophobic bonds of existing antimicrobial materials require long dehydration times before they become effective, during which significant microbial transmission occurs, and therefore the antimicrobial activity is not effective against hardy microorganisms (such as Escherichia coli (E. Coli)).

[0004] Thus, there is a need for antimicrobial compounds and nanostructures that enable further functionalization and stronger antimicrobial activity against microorganisms (such as E. Coli), compositions comprising the compounds and nanostructures, and methods using the compounds, nanostructures and compositions useful for antimicrobial and / or antiviral applications. Summary of the Invention [Problem to be solved by the invention]

[0005] The present disclosure provides compounds and nanostructures having one or more quaternary ammonium salts, compositions comprising the compounds and nanostructures, and methods useful for treating conditions, such as conditions associated with viruses and toxins, using the compounds, nanostructures and compositions. [Means for solving the problem]

[0006] At least one compound has the formula (I): [ka] or a pharma- ceutical acceptable salt thereof, Q is fluoro, chloro, bromo or iodo; each of s, b, and n is independently an integer of about 10 to about 100; Each of v, j, p, z, q, x and m is independently an integer of 1 to about 20.

[0007] At least one composition of the present disclosure comprises a compound of formula (I) or a pharma- ceutically acceptable salt thereof and one or more additional components. The composition may have a three-dimensional shape that is a nanoworm or a nanorod.

[0008] At least one method includes depositing a compound or composition of the present disclosure onto an object, such as an interior surface of an aircraft.At least one method includes a method for treating a condition, comprising administering to a subject a therapeutically effective amount of a compound represented by Formula (I) or a pharma- ceutically acceptable salt thereof (or a composition comprising a compound represented by Formula (I) or a pharma- ceutically acceptable salt thereof), wherein the condition being treated includes a viral infection, a bacterial infection, a chronic inflammatory disorder, an acute inflammatory disorder, or cancer.

[0009] At least one method includes preparing a compound of formula (I) or a pharma- ceutically acceptable salt thereof.At least one method includes preparing a composition comprising a compound of formula (I) or a pharma- ceutically acceptable salt thereof.

[0010] So that the above-mentioned features of the present disclosure may be understood in detail, the present disclosure, briefly summarized above, can be further described by reference to embodiments, some of which are illustrated in the accompanying drawings. It should be noted, however, that the accompanying drawings illustrate only typical embodiments of the present disclosure, and therefore should not be considered as limiting the scope thereof, since the present disclosure recognizes other embodiments that are equally valid. [Brief description of the drawings]

[0011] [Figure 1] FIG. 2 shows a 1H nuclear magnetic spectrum of a polymer in deuterated chloroform according to one embodiment. [Figure 2A] FIG. 2 shows a 1H NMR spectrum of maleimide-functionalized quaternized polyDMAEA in deuterium oxide according to one embodiment. [Figure 2B] FIG. 2 shows a 1H NMR spectrum of maleimide-functionalized quaternized polyDMAEA in deuterium oxide according to one embodiment. [Figure 2C] FIG. 2 shows a 1H NMR spectrum of maleimide-functionalized quaternized polyDMAEA in deuterium oxide according to one embodiment. [Diagram 3] FIG. 2 shows a scheme illustrating the conjugation of maleimide-functionalized cationic polymers with Dopa- and PDS-functionalized nanoworms, according to one embodiment. [Figure 4] 1 is a bar graph showing the inhibitory effect of several polymers of the present disclosure on E. coli culture, according to one embodiment. [Figure 5-1] 5A and 5B are scanning electron microscope images showing the antibacterial activity of several cationic polymer nanoworms of the present disclosure against E. coli, according to one embodiment. [Figure 5-2]5C and 5D are scanning electron microscopy images showing the antimicrobial activity of several cationic polymer nanoworms of the present disclosure against E. coli, according to one embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] For ease of understanding, the same reference numbers have been used, where possible, to designate identical elements common to the figures. The figures are not drawn to scale and may be simplified for clarity. It is contemplated that elements and features of one embodiment may be beneficially incorporated in other embodiments without further recitation.

[0013] The present disclosure provides compounds and nanostructures that include one or more quaternary ammonium salts, compositions that include the compounds and nanostructures, and methods useful for treating conditions using the compounds, nanostructures and compositions.

[0014] compound In at least one embodiment, the compound has formula (I): [ka] or a pharma- ceutically acceptable salt thereof, wherein Q is fluoro, chloro, bromo or iodo, preferably chloro; each of s, b, and n is independently an integer of about 10 to about 100; Each of v, j, p, z, q, x and m is independently an integer of 1 to about 20.

[0015] In at least one embodiment, s can be an integer from about 20 to about 40, such as from about 25 to about 35. b can be an integer from about 30 to about 60, such as from about 40 to about 50. n can be an integer from about 30 to about 60, such as from about 50 to about 60. m can be an integer from about 1 to about 10, such as from about 5 to about 10. x can be an integer from about 5 to about 15, for example, 7, 11, or 15.

[0016] Without being limited by theory, an x ​​value of about 5 to about 15 provides cell membrane penetration of the alkyl moiety into the hydrophobic portion of the cell membrane (such as a viral cell), while the cationic nitrogen moiety of the compound of formula (I) provides Coulombic interaction of the compound of formula (I) (such as a quaternary ammonium moiety) with the cell membrane surface (such as the phosphate ester portion of a phospholipid bilayer). Additionally, the quaternary ammonium salt provides sufficient hydrophilicity such that the alkyl moiety attached to the quaternary ammonium salt is not significantly buried within the core of the three-dimensional structure (e.g., when the composition has a nanoworm or nanorod three-dimensional structure).

[0017] The polystyrene block of the compound of formula (I) can provide the compound of formula (I) with a high glass transition temperature (Tg) component (100% polystyrene has a Tg of about 100°C). The high Tg provides stability to the nanostructures at body temperature. Furthermore, the poly(N-isopropylacrylamide) (polyNIPAM) block can provide the compound of formula (I) with a nanoworm (or nanorod) three-dimensional shape under aqueous conditions, for example, in an aqueous solution containing sodium dodecyl sulfate (SDS).

[0018] The ratio of integer n to integer m can be selected to fine-tune the coulombic binding ability to the cell membrane permeability of the compound of formula (I). A larger value of integer n increases the coulombic binding, and a larger value of integer m increases the cell membrane permeability of the compound of formula (I). In at least one embodiment, the ratio of integer n to integer m is about 1:1 to about 100:1, such as about 5:1 to about 15:1, for example 54:7.

[0019] The values ​​of v, j, p, z, and q can be varied based on the number of, e.g., methylene units, of the starting materials used to generate the compound of formula (I). The value of s can be controlled by controlling the molar ratio of styrene monomer to NIPAM monomer and maleimide starting materials, as described in more detail below. Similarly, the value of b can be controlled by controlling the molar ratio of NIPAM monomer to styrene monomer and maleimide starting materials. The values ​​of n and m can be controlled by the ratio of, e.g., methyl halide to other alkyl halides (e.g., octyl halide) used to form the quaternary ammonium salt moiety.

[0020] In at least one embodiment, the compound of formula (I) has formula (II): [ka] or a pharma- ceutically acceptable salt thereof, wherein Q is fluoro, chloro, bromo or iodo, preferably chloro; each of s, b, and n is independently an integer of about 10 to about 100; Each of x and m is independently an integer of 1 to about 20.

[0021] In at least one embodiment, s in formula (II) can be an integer from about 20 to about 40, such as from about 25 to about 35. b in formula (II) can be an integer from about 30 to about 60, such as from about 40 to about 50. n in formula (II) can be an integer from about 30 to about 60, such as from about 50 to about 60. m in formula (II) can be an integer from about 1 to about 10, such as from about 5 to about 10. x in formula (II) can be an integer from about 5 to about 15.

[0022] In at least one embodiment, the compound of formula (I) or (II) has formula (III): [ka] or a pharma- ceutically acceptable salt thereof (wherein x is an integer of about 5 to about 15, for example, 7, 11, or 15).

[0023] Composition: The compound of formula (I), (II) or (III) or a pharma- ceutically acceptable salt thereof may be present in a composition that includes one or more additional ingredients, which may include one or more pharma- ceutically active compounds.

[0024] In at least one embodiment, the composition comprises a compound having formula (I): [ka] or a pharma- ceutical acceptable salt thereof, wherein Q is fluoro, chloro, bromo or iodo, preferably chloro; each of s, b, and n is independently an integer of about 10 to about 100; Each of v, j, p, z, q, x and m is independently an integer of 1 to about 20.

[0025] In at least one embodiment, s can be an integer from about 20 to about 40, such as from about 25 to about 35. b can be an integer from about 30 to about 60, such as from about 40 to about 50. n can be an integer from about 30 to about 60, such as from about 50 to about 60. m can be an integer from about 1 to about 10, such as from about 5 to about 10. x can be an integer from about 5 to about 15, for example, 7, 11, or 15.

[0026] In at least one embodiment, the ratio of integer n to integer m is from about 1:1 to about 100:1, such as from about 5:1 to about 15:1, for example, 54:7.

[0027] In at least one embodiment, the compound of formula (I) has formula (II): [ka] or a pharma- ceutically acceptable salt thereof, wherein Q is fluoro, chloro, bromo or iodo, preferably chloro; each of s, b, and n is independently an integer of about 10 to about 100; Each of x and m is independently an integer of 1 to about 20.

[0028] In at least one embodiment, s in formula (II) can be an integer from about 20 to about 40, such as from about 25 to about 35. b in formula (II) can be an integer from about 30 to about 60, such as from about 40 to about 50. n in formula (II) can be an integer from about 30 to about 60, such as from about 50 to about 60. m in formula (II) can be an integer from about 1 to about 10, such as from about 5 to about 10. x in formula (II) can be an integer from about 5 to about 15.

[0029] In at least one embodiment, the compound of formula (I) or (II) has formula (III): [ka] or a pharma- ceutically acceptable salt thereof (wherein x is an integer of about 5 to about 15, for example, 7, 11, or 15).

[0030] The compositions of the present disclosure further comprise a compound of formula (IV): [ka] or a pharma- ceutical acceptable salt thereof (wherein, in formula (IV), s and b each independently represent an integer of about 10 to about 100; In formula (IV), v is an integer of about 1 to about 20; R 1 is -O- or -NH-, R 2 is -CH3, biotin, pyridyl disulfide, dopa, thiolactone, or adamantyl. Biotin can be of the structure: [ka] The pyridyl disulfide can be of the structure: [ka] Dopa can be of the following structure: [ka] Thiolactone is [ka] Adamantyl can be a γ-thiolactone of the structure [ka] The structure may be as follows:

[0031] R as pyridyl disulfide 2 are particularly advantageous since they can be further conjugated to a pharma- ceutical active moiety. Pharmaceutically active moieties include polymers, sugars, peptides, oligonucleotides, proteins, or small molecule (e.g., 2,000 g / mol or less) therapeutics, such as anti-cancer drugs. Proteins include glycoproteins, such as Vitronectin. Small molecule therapeutics include Relenza binding agents. Peptides include twin-arginine or translocation (TAT) peptide. Oligonucleotides include siRNA.

[0032] Additionally, pyridyl disulfides can undergo thiol-disulfide exchange or thiol-ene reactions, Dopa can be attached to metal surfaces, γ-thiolactones can undergo amination reactions (e.g., with amino acids), biotin can be attached to biomolecules (e.g., streptavidin), and adamantly can be brominated, fluorinated, carboxylated, or hydroxylated.

[0033] In at least one embodiment, s in formula (IV) is an integer of from about 25 to about 35. b in formula (IV) can be an integer of from about 40 to about 50. v in formula (IV) can be an integer of from 1 to about 10, for example, 2.

[0034] In at least one embodiment, the compound of formula (IV) is one or more of the following compounds of formula (V), formula (VI) or formula (VII) or a pharma- ceutically acceptable salt thereof: [ka] (wherein s is an integer of about 25 to about 35, and b is an integer of about 40 to about 50).

[0035] In at least one embodiment, the ratio of the compound represented by Formula (I), (II) or (III) to the compound represented by Formula (IV), Formula (V), Formula (VI) or Formula (VII) is from about 0.01:1 to about 1:0.01.

[0036] In at least one embodiment, the composition includes a compound represented by formula (I), (II) or (III), a compound represented by formula (V), and an additional compound represented by formula (IV). The ratio of the compound represented by formula (I), (II) or (III) to the compound represented by formula (V) to the compound represented by formula (IV) is about 0.99:0.005:0.005 to about 0.01:0.09:0.009, such as about 0.9:0.05:0.05 to about 0.1:0.08:0.01, such as about 0.8:0.1:0.1 to about 0.1:0.1:0.8.

[0037] Three-dimensional structure of a compound or composition The long, flexible worms have an in vivo blood circulation time approximately 10 times longer than their spherical analogues, and a circulation time significantly longer than any synthetic particle or polyethylene oxide-coated vesicle, whereas the short rods have a much shorter circulation time but are taken up by cells more efficiently.

[0038] The compounds or compositions of the present disclosure may have a three-dimensional structure that is a nanoworm or nanorod. The nanorod may have an aspect ratio of about 10:1 to about 1000:1, such as about 25:1 to about 75:1, such as about 10:1 to about 100:1. The nanorod may have a diameter of about 10 nm to about 20 nm and a length of about 100 nm to about 10 microns, such as about 1 micron to about 2 microns.

[0039] The compounds and compositions of the present disclosure may also have a three-dimensional structure that is a sphere, a vesicle, a donut, or a lamellar sheet. The three-dimensional structure of the compositions of the present disclosure may be stable in water for long periods (e.g., nanoworms are stable at room temperature for more than one year), and can be lyophilized and rehydrated without structural reorganization. For example, nanoworm solutions can be lyophilized to obtain dry power. The lyophilized product can be rehydrated in Milli-Q water at 8 wt% for 2 hours. The ability of the compositions of the present disclosure to be lyophilized allows the compositions of the present disclosure to be transported stably.

[0040] Nanoworm Generation Preparation of functional thermoresponsive polymer nanorods: RAFT-mediated polymerization of styrene using functional poly(NIPAM) macro-chain transfer agent (macro-CTA) in water: A precursor compound (e.g., a compound represented by formula (I) but not including a polystyrene block) can be mixed with styrene, sodium dodecyl sulfate, and azobisisobutyronitrile (AIBN) in water (such as Milli-Q water) to produce a reaction mixture. One or more additional precursor compounds (e.g., a compound represented by formula (IV) but not including a polystyrene block) can also be added to the reaction mixture.

[0041] AIBN can be dissolved in styrene and then the solution can be added to a flask containing the other reaction ingredients to produce a reaction mixture. The solution of AIBN and styrene can be deoxygenated by purging with argon for, e.g., about 15 minutes. The reaction mixture can be purged with argon in an ice bath for, e.g., about 10 minutes, and then heated to 70° C. for about 3.5 hours to produce a product mixture.

[0042] The number average molecular weight (Mn) of the compound of formula (IV) in the product mixture is determined by nuclear magnetic resonance spectroscopy unless otherwise stated. 1 H NMR can be measured by lyophilizing the product mixture to remove all water and low boiling compounds, then dissolving the product mixture in CDCl3. Size exclusion chromatography spectra can be obtained by taking 3 drops of the latex, dissolving in 1 mL of THF, filtering, and injecting the solution into the SEC instrument.

[0043] Alternatively, the conversion of the styrene monomer can be determined gravimetrically. The theoretical molecular weight can be calculated based on the monomer conversion. Molecular weight (M n(SEC) ) and PDI SEC can be obtained by THF SEC. n(NMR) )teeth 1 Obtained from H NMR. Particle size (D h ) and particle size distribution (PDI DLS ) could be measured by dynamic light scattering (DLS) at 70 °C immediately after the reaction was stopped, and the results were the average values ​​based on three measurements. PDI less than 0.1 DLS means that the distribution is narrow.

[0044] Nanoworms cut into nanorods using ultrasound Nanorods can be obtained by temperature directed morphology transformation (TDTM) and ultrasonic cutting of nanoworms. In at least one embodiment, 6 mL of latex solution of nanoworms can be transferred into two hot vials (3 mL each) with 60SL of toluene in each vial. These vials can then be sealed and shaken. The suspension in these vials can be cooled to 23°C. The latex solution can be cooled from 70°C to 15°C for about 30 minutes. To confirm the generation of worm-like nanostructures, the nanostructures can be characterized by transmission electron microscopy (TEM). To generate rods, the worms can be diluted by adding 10 mL of Milli-Q water and cut in an ice bath with an ultrasonic probe (pulsed 3 seconds on and 2 seconds off as one pulse cycle) at 35% power for 3 minutes (3 mm tapered microtip, VC-750 system from Sonics&Materials). After ultrasonic cutting, the nanostructures can be characterized again by TEM to confirm the generation of rods.

[0045] Ultrasonic cutting of nanoworms into nanorods can also be performed by applying probed ultrasound with different pulse cycles (15 s on and 10 s off as one pulse cycle), (B) 12 cycles (3 min), (C) 36 cycles (9 min) and (D) 48 cycles (12 min).

[0046] In at least one embodiment, the nanoworm or nanorod compositions of the present disclosure can be heated above the lower critical solution temperature (LCST) of the PNIPAM block (about 37° C.) to form a gel, which dissociates back into a sol when cooled, i.e., a reversible process. Nanoworms can form gels at a minimum weight fraction of about 1 wt % to about 8 wt % in aqueous solution. Nanorods can form gels at a minimum weight fraction of about 2 wt % to about 16 wt % in aqueous solution. Without being limited by theory, gels are advantageous because they dissociate at elevated temperatures (e.g., from room temperature to the body temperature of a subject, such as a human), allowing dissociation of the worm three-dimensional structure and movement through the blood.

[0047] The weight percent of nanorods in water that can produce a gel at 37° C. can be determined as follows: In general, lyophilized nanorods (e.g., 20 mg) can be redispersed in Milli-Q water by vortexing in a 1.5 ml Eppendorf tube at 25° C. at 30 wt%. The tube can then be capped and immersed in a 37° C. water bath for 2 minutes. The tube can then be inverted under the water bath and observed for gel formation. Gel formation is defined as no fluid flow being observed within 30 seconds. The weight percent can be lowered by adding more Milli-Q water and vortexing. Gel formation can then be checked again. The minimum weight percent of nanorods (e.g., in water) that will produce a gel at 37° C. is defined as the wt% that will produce a gel.

[0048] Preferred Methods for the Synthesis of Compounds of Formula (I) A preferred method for synthesizing a compound of formula (I) is shown in Scheme 1. A pyridyl disulfide-capped diblock copolymer is reacted with a maleimide-capped quaternary ammonium polymer in the presence of tris(2-carboxyethyl)phosphine (TCEP) in water to produce a compound of formula (I). The pyridyl disulfide-capped diblock copolymer can be a compound of formula (IV) or (V) as the starting material to be reacted with a maleimide-capped quaternary ammonium polymer to produce a compound of formula (I). Scheme 1 [ka]

[0049] Methods for depositing compounds and compositions The compounds and compositions of the present disclosure may be deposited on the surface of an object by any suitable deposition method.Deposition methods may include one or more of painting, dipping, spraying, marking, taping, brushing, spin coating, roll coating, doctor blade coating.Before deposition, the compounds or compositions of the present disclosure may be diluted in a solvent such as water.After deposition, the solvent may then evaporate at room temperature to form a compound / composition layer on the object.

[0050] In at least one embodiment, the object is an interior surface of an aircraft / spacecraft / boat or an air filter surface of an aircraft / spacecraft / boat, such as the surface of an air conditioning or filtration system.The object can be the interior floor surface, seat surface, overhead bin surface, ceiling surface, door surface and / or door handle surface of an aircraft.

[0051] In at least one embodiment, the compound or composition of the present disclosure is sprayed onto the surface of the object for about 1 second to about 10 minutes, such as about 30 seconds to about 2 minutes. In at least one embodiment, the compound or composition is sprayed onto the surface of the object in an amount of about 1 mL to about 25 kL, such as about 100 L to about 1 kL.

[0052] The compounds or compositions of the present disclosure disposed on an object can prevent, reduce and / or eliminate the presence of bacteria and viruses, thereby preventing, reducing and / or eliminating human contact with such bacteria and viruses.

[0053] Compositions comprising the nanostructures (e.g., nanorods or nanoworms) of the present disclosure are advantageous for deposition on a surface, for example, because antibacterial and antiviral compounds can be applied as a monolayer, maintaining the efficacy of both compounds. As described above, for example, a composition comprising a conventional antimicrobial compound can be applied to a surface to form a first layer on the surface. For example, a composition comprising a conventional antiviral compound can be applied to the first layer to form a second layer. However, the second layer (antiviral layer) will mask (hinder the antimicrobial capabilities of) the antimicrobial first layer.

[0054] Applying the nanostructured composition as a single layer also reduces the cost and time of applying the compound to a surface as compared to applying two or more layers.

[0055] Methods for use as a medicine The present disclosure further provides a method for treating a condition in a subject who is in such a condition or is susceptible to such a condition by administering to the subject a therapeutically effective amount of one or more of the above-mentioned compounds or compositions.In one embodiment, the treatment is a preventive treatment.In another embodiment, the treatment is a symptomatic treatment.In another embodiment, the treatment is a restorative treatment.

[0056] A method for treating a condition can include administering to a subject a therapeutically effective amount of a compound represented by formula (I) or a pharma- ceutical acceptable salt thereof (or a composition comprising a compound represented by formula (I) or a pharma- ceutical acceptable salt thereof).

[0057] 1. Status Conditions that can be treated according to the present disclosure include, but are not limited to, conditions caused by toxins (such as antigens) and inflammatory disorders such as septic shock. Conditions that can be treated according to the present disclosure include, but are not limited to, viral infections, bacterial infections, chronic inflammatory disorders, acute inflammatory disorders, and cancer. Preferably, the conditions treated include bacterial infections, viral infections, or cancer immunotherapy. Cancer immunotherapy can include cervical cancer, for example, resulting from infection of the cervix by human papillomavirus.

[0058] Viral infections may include those caused by Ebola, influenza, SARS, norovirus (gastric) or Zika. Viral infections may include viral respiratory infections (e.g., of the nose, pharynx, upper respiratory tract or lungs), such as pneumonia, laryngotracheobronchitis, bronchiolitis. Viral infections may include viral gastrointestinal infections, such as gastroenteritis caused by norovirus or rotavirus. Viral infections may include viral liver infections, such as hepatitis. Viral infections may include viral nervous system infections, such as encephalitis caused by rabies or West Nile. Viral infections include warts and / or infections caused by human papillomavirus (HPV). Viral infections may include infections that cause cancer, such as infections caused by Epstein-Barr virus, hepatitis B, hepatitis C, herpes virus 8 or human papillomavirus. Symptoms of a viral infection may include fever, muscle aches, cough, sneezing, runny nose, headache, chills, diarrhea, vomiting, rash or weakness.

[0059] Bacterial infections may include pneumonia, meningitis, food poisoning and bacterial skin infections, such as those caused by Staphylococcus or Streptococcus, cellulitis, folliculitis, impetigo and boils. Bacterial infections (e.g., from food poisoning) may include infections caused by E. coli, Campylobacter jejuni, Clostridium botulinum, Listeria monocytogenes, Salmonella and Vibrio. Bacterial infections may include bacterial meningitis, otitis media, urinary tract infections, and respiratory tract infections, such as pharyngitis, bronchitis, sinusitis and pneumonia. Symptoms of bacterial infections may include nausea, vomiting, diarrhea, fever, chills and abdominal pain.

[0060] In some embodiments, the methods described herein are used to treat patients with disorders caused by dysregulated cytokine, enzyme and / or inflammatory mediator production, stability, secretion, post-translational processing.Examples of cytokines that can be dysregulated include interleukin 1, 2, 6, 8, 10, 12, 17, 22 and 23, along with tumor necrosis factor alpha and interferon alpha, beta and gamma.Examples of inflammatory mediators that can be dysregulated include nitric oxide, prostaglandins and leukotrienes.Examples of enzymes include cyclooxygenase, nitric oxide synthase and matrix metalloprotease.

[0061] Examples of inflammatory conditions relevant to the present technology 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 immunosuppressed individuals and may also include "superbugs," including bacterial and viral strains that are resistant to current therapies.

[0062] 2. Target Suitable subjects to be treated according to the present disclosure include mammalian subjects. Mammals according to the present disclosure include, but are not limited to, humans, dogs, cats, cows, goats, horses, sheep, pigs, rodents, lagomorphs, primates, etc., including mammals in utero. Subjects may be of either sex and at any stage of development.

[0063] 3. Administration and Dosing The compounds or compositions of the disclosure can be administered to a subject in a therapeutically effective amount.

[0064] The compounds or compositions of the present disclosure can be administered by any suitable route, in the form of a pharmaceutical composition adapted to such route, and in a dose effective for the intended treatment. The effective dose is typically within the range of about 0.001 to about 100 mg / kg body weight / day, preferably about 0.01 to about 30 mg / kg / day, in a single dose or in divided doses. Depending on the age, species and condition being treated, dosage levels below the lower end of this range may be appropriate. In some cases, even higher doses can be used without side effects. The higher dose can also be divided into several lower doses for administration throughout the day.

[0065] Pharmaceutical Compositions For the treatment of the above-mentioned conditions, the compounds described herein can be administered as follows.

[0066] Oral route A compound or composition of the disclosure can be administered orally, including by swallowing, so that the compound enters the digestive tract or is absorbed into the bloodstream directly from the mouth (e.g., buccal or sublingual administration).

[0067] 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 or modified release, including delayed or sustained release, optionally with an enteric coating.

[0068] Liquid preparations may include solutions, syrups and suspensions, which can be used in soft or hard capsules.Such preparations may include pharma-ceutically acceptable carriers, such as water, ethanol, polyethylene glycol, cellulose or oil.The preparations may also include one or more emulsifiers and / or suspending agents.

[0069] In tablet dosage forms, the amount of the compound of formula (I) present may be from about 0.05% to about 95% by weight, such as from about 2% to about 50% by weight of the dosage form. Additionally, the tablet may contain a disintegrant comprising from about 0.5% to about 35% by weight, such as from about 2% to about 25% by weight of the dosage form. Examples of disintegrants include methylcellulose, sodium or calcium carboxymethylcellulose, croscarmellose sodium, polyvinylpyrrolidone, hydroxypropylcellulose or starch.

[0070] Lubricants suitable for use in tablets may be present in an amount of about 0.1% to about 5% by weight and may include calcium, zinc or magnesium stearate or sodium stearyl fumarate.

[0071] Binders suitable for use in tablets include gelatin, polyethylene glycol, sugars, gums, starch, hydroxypropylcellulose, etc. Diluents suitable for use in tablets include mannitol, xylitol, lactose, dextrose, sucrose, sorbitol or starch.

[0072] Surfactants and glidants suitable for use in tablets may be present in amounts of about 0.1% to about 3% by weight. Surfactants and glidants can include polysorbate 80, sodium dodecyl sulfate, talc, or silicon dioxide.

[0073] Parenteral Administration The compounds and compositions of the present disclosure can be administered directly into the bloodstream, into muscle or into internal organs.Methods suitable for parenteral administration can include intravenous, intramuscular, subcutaneous, intraarterial, intraperitoneal, intrathecal or intracranial.Devices suitable for parenteral administration include syringes (including needle and needleless syringes) and infusion methods.

[0074] Compositions for parenteral administration may be formulated as immediate or modified release, including delayed or sustained release.

[0075] Most parenteral formulations are aqueous solutions containing excipients, including salts, buffers and carbohydrates.

[0076] Parenteral formulations can also be prepared in a dehydrated form (e.g., by lyophilization) or as a non-aqueous sterile solution. These formulations can contain water. Solubility enhancers can also be used in the preparation of parenteral solutions.

[0077] Topical administration The compounds and compositions of the present disclosure can be administered topically or transdermally to the skin.The topical formulations can include lotions, solutions, creams, gels, hydrogels, ointments, foams, implants, patches, etc.The pharma- ceutically acceptable carriers for topical formulations can include water, alcohol, mineral oil, glycerin, polyethylene glycol, etc.Topical administration can be performed by electroporation, iontophoresis, or phonophoresis.

[0078] Compositions for topical administration may be formulated as immediate or modified release, including delayed or sustained release.

[0079] Combinations and combination therapies The compounds and compositions of the present disclosure can be used alone or in combination with other medicament active compounds to treat conditions such as those previously mentioned above.The compound(s) / composition(s) of the present disclosure and the other medicament active compound(s) can be administered simultaneously (either in the same dosage form or in separate dosage forms) or sequentially.Thus, in one embodiment, the present disclosure includes a method for treating a condition by administering to a subject a therapeutically effective amount of one or more compounds of the present disclosure and one or more additional medicament active compounds.

[0080] In another aspect, there is provided a pharmaceutical composition comprising one or more compounds of the present disclosure, one or more additional pharma- ceutically active compounds, and a pharma- ceutically acceptable carrier.

[0081] In another embodiment, the one or more additional pharma- ceutical active compounds are one or more anti-inflammatory agents, anti-atherosclerotic agents, immunosuppressants, immunomodulatory agents, cytostatic agents, antiproliferative agents, angiogenesis inhibitors, kinase inhibitors, cytokine blockers, or inhibitors of cell adhesion molecules.

[0082] The compounds and compositions of the present disclosure can also be used in combination with other therapeutic agents that are selected for their therapeutic value against the condition being treated. In general, the compounds and compositions described herein, as well as other agents in the embodiment in which combination therapy is used, do not need to be administered in the same pharmaceutical composition, and are optionally administered by different routes due to their different physical and chemical properties. Initial administration is generally performed according to an established protocol, and then the dosage, method of administration, and number of administrations are subsequently modified based on the observed effects. In certain instances, it is appropriate to administer the compounds of formula (I) described herein in combination with another therapeutic agent. By way of example only, if one of the side effects experienced by the patient when receiving the compound of formula (I) is a rash, it is appropriate to administer an antihistamine in combination with the initial therapeutic agent. Or, by way of example only, the therapeutic effectiveness of the compound of formula (I) is improved by the administration of another therapeutic agent (including a treatment regimen) that also has a therapeutic benefit. Regardless of the disease, disorder, or condition being treated, the overall benefit experienced by the patient is simply the additive of the two therapeutic agents or the patient experiences a synergistic benefit.

[0083] When drugs are used in combination therapy, therapeutically effective dosages vary. Methods for experimentally determining therapeutically effective dosages of drugs and other agents for use in combination therapy regimes are described techniques. Combination therapy also includes periodic therapy that starts and stops at different times to assist in the clinical management of patients. In any case, multiple therapeutic agents (one of which is the compound of formula (I)) are administered in any order or even simultaneously. When simultaneously, multiple therapeutic agents are optionally provided in a single integrated form or multiple forms (for example only, as one pill or two separate pills).

[0084] In some embodiments, one of the therapeutic agents is given in multiple doses, or both are given as multiple doses. If not simultaneous, the timing between the multiple doses optionally varies from more than 0 weeks to less than 12 weeks.

[0085] Moreover, combination methods, compositions and formulations should not be limited to the use of only two drugs, and the use of multiple therapeutic combinations is also envisioned.It is understood that the dosage regimen for treating, preventing or ameliorating the condition(s) that is sought to be alleviated is optionally modified according to various factors.These factors include the disorder that the subject suffers from, as well as the subject's age, weight, sex, diet and medical condition.Therefore, the dosage regimen that is actually used may differ from the dosage regimen described herein, since it may vary widely in some aspects.

[0086] The pharmaceutical agents constituting the combination therapy disclosed herein are optionally in combined dosage forms or separate dosage forms intended for substantially simultaneous administration.The pharmaceutical agents constituting the combination therapy are also optionally administered sequentially, with either agent being administered by a dosing regimen requiring two-step administration.The two-step dosing regimen optionally requires sequential administration of active agents or separate, spaced administration of active agents.The time between the multiple administration steps ranges from minutes to hours, depending on the properties of each pharmaceutical agent, such as the pharmaceutical agent's potency, solubility, bioavailability, plasma half-life and kinetic profile.The circadian variation of the target molecule concentration is optionally used to determine the optimal administration interval.

[0087] The compound of formula (I) and the composition comprising the compound of formula (I) can be used (e.g., administered) in combination with drugs from the following classes: NSAIDs, immunosuppressants, immunomodulators, cytostatics, antiproliferatives, angiogenesis inhibitors, biological agents, steroids, vitamin D3 analogs, retinoids, other kinase inhibitors, cytokine blockers, corticosteroids and inhibitors of cell adhesion molecules. If a subject suffers from or is at risk of suffering from atherosclerosis or a condition associated with atherosclerosis, the compound of formula (I) or the composition comprising the compound of formula (I) described herein is optionally used 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 treating atherosclerosis or a condition associated with atherosclerosis include, but are not limited to, any of the following: torcetrapib, aspirin, niacin, HMG CoA reductase inhibitors (e.g., atorvastatin, fluvastatin, lovastatin, pravastatin, rosuvastatin, and simvastatin), colesevelam, cholestyramine, colestipol, gemfibrozil, probucol, and clofibrate.

[0088] When a subject suffers from or is at risk of suffering from an inflammatory condition, a compound of formula (I) or a composition comprising a compound of formula (I) described herein is optionally used in any combination with one or more agents or methods for treating the inflammatory condition. Examples of therapeutic agents / treatments for treating autoimmune and / or inflammatory conditions include, but are not limited to, any of the following: corticosteroids, nonsteroidal anti-inflammatory drugs (NSAIDs) (e.g., ibuprofen, naproxen, acetaminophen, aspirin, fenoprofen (Nalfon), flurbiprofen (Ansaid), ketoprofen, oxaprozin (Daypro), diclofenac sodium (Voltaren), diclofenac potassium (Cataflam), etodolac (Lodine), indomethacin (Indocin), ketorolac (Toradol), sulindac (Clinoril), tolmetin (Tolectin), meclofenamate (Meclomen), mefenamic acid (Ponstel), nabumetone (Relafen), piroxicam (Feldene), cox-2 inhibitors (e.g., celecoxib (C elebrex)), 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) and adalimumab (Humira)), abatacept (CTLA4-Ig) and interleukin-1 receptor antagonists (e.g., anakinra (Kineret), interleukin-6 inhibitors (e.g., Actemra), interleukin-17 inhibitors (e.g., AlN457), Janus kinase inhibitors (e.g., Tasocitinib), syk inhibitors (e.g., R788), chloroquine and its derivatives.

[0089] For use in cancer and neoplastic diseases, the compounds of formula (I) or compositions comprising compounds of formula (I) described herein are optionally used with one or more of the following classes of drugs: anti-cancer drugs are EGFR kinase inhibitors, MEK inhibitors, VEGFR inhibitors, anti-VEGFR2 antibodies, KDR antibodies, AKT inhibitors, PDK-1 inhibitors, PI3K inhibitors, c-kit / Kdr tyrosine kinase inhibitors, Bcr-Abl tyrosine kinase inhibitors, VEGFR2 inhibitors, PDGFR-beta inhibitors, KIT inhibitors, Flt3 tyrosine kinase inhibitors, PDGF receptor family inhibitors, Flt3 tyrosine kinase inhibitors, RET tyrosine kinase receptor family inhibitors, VEGF-3 receptor antagonists, Raf protein kinase family inhibitors, angiogenesis inhibitors, Erb2 inhibitors, mTOR inhibitors, IGF-1R antibodies, NFkB inhibitors, proteosome inhibitors, chemotherapeutic agents or glucose reducing agents. Aspects Clause 1. Formula (I): [ka] or a pharma- ceutical acceptable salt thereof (wherein Q is fluoro, chloro, bromo or iodo; each of s, b, and n is independently an integer of about 10 to about 100; Each of v, j, p, z, q, x and m is independently an integer from 1 to about 20. Clause 2. The compound according to clause 1, wherein s is an integer from about 20 to about 40. Clause 3. The compound according to clause 1 or 2, wherein s is an integer from about 25 to about 35. The compound according to any one of clauses 1 to 3, wherein clause 4.b is an integer from about 30 to about 60. The compound according to any one of clauses 1 to 4, wherein clause 5.b is an integer from about 40 to about 50. Clause 6. The compound according to any one of clauses 1 to 5, wherein n is an integer from about 30 to about 60. Clause 7. The compound according to any one of clauses 1 to 6, wherein n is an integer from about 50 to about 60. Clause 8. The compound according to any one of clauses 1 to 7, wherein m is an integer from about 1 to about 10. Clause 9. The compound according to any one of clauses 1 to 8, wherein m is an integer from about 5 to about 10. Clause 10. The compound according to any one of clauses 1 to 9, wherein Q is chloro. Clause 11. The compound according to any one of clauses 1 to 10, wherein x is an integer from about 5 to about 15. Clause 12. The compound according to any one of clauses 1 to 11, wherein x is an integer 7, 11 or 15. Clause 13. The compound according to any one of clauses 1 to 12, wherein the ratio of integer n to integer m is from about 1:1 to about 100:1. Clause 14. The compound according to any one of clauses 1 to 13, wherein the ratio of integer n to integer m is from about 5:1 to about 15:1. Article 15. Formula (II): [ka] or a pharma- ceutical acceptable salt thereof, Q is fluoro, chloro, bromo or iodo; each of s, b, and n is independently an integer of about 10 to about 100; Each of x and m is independently an integer from 1 to about 20. Clause 16. The compound according to clause 15, wherein s in formula (II) is an integer of about 20 to about 40. Clause 17. The compound according to clause 15 or 16, wherein s in formula (II) is an integer of about 25 to about 35. Clause 18. The compound according to any one of clauses 15 to 17, wherein b in formula (II) is an integer of about 30 to about 60. Clause 19. The compound according to any one of clauses 15 to 18, wherein b in formula (II) is an integer of about 40 to about 50. Clause 20. The compound according to any one of clauses 15 to 19, wherein n in formula (II) is an integer of about 30 to about 60. Clause 21. The compound according to any one of clauses 15 to 20, wherein n in formula (II) is an integer of about 50 to about 60. Clause 22. The compound according to any one of clauses 15 to 21, wherein m in formula (II) is an integer of from about 1 to about 10. Clause 23. The compound according to any one of clauses 15 to 22, wherein m in formula (II) is an integer of about 5 to about 10. Clause 24. The compound according to any one of clauses 15 to 23, wherein Q is chloro. Clause 25. The compound according to any one of clauses 15 to 24, wherein x in formula (II) is an integer of about 5 to about 15. Article 26. Formula (III): [ka] 26. The compound according to any one of clauses 15 to 25, represented by the formula: (wherein x is an integer of about 5 to about 15) or a pharma- ceutically acceptable salt thereof. Clause 27. The compound according to any one of clauses 15 to 26, wherein x is an integer 7, 11 or 15. Clause 28. A compound according to any one of clauses 15 to 27, Formula (IV): [ka] or a pharma- ceutical acceptable salt thereof (wherein In formula (IV), s and b each independently represent an integer of about 10 to about 100; In formula (IV), v is an integer of about 1 to about 20; R 1 is -O- or -NH-, R 2 is -CH3, biotin, pyridyl disulfide, dopa, thiolactone or adamantyl. and a composition comprising: Clause 29. The composition according to clause 28, wherein s in formula (IV) is an integer of about 25 to about 35. Clause 30. The composition according to clause 28 or 29, wherein b in formula (IV) is an integer of about 40 to about 50. Clause 31. The composition according to any one of clauses 28 to 30, wherein v in formula (IV) is 2. Article 32.R 2 but [ka] 32. The composition according to any one of clauses 28 to 31, wherein Clause 33. The compound of formula (IV) [ka] wherein s in formula (IV) is an integer of about 25 to about 35, and b in formula (IV) is an integer of about 40 to about 50. 33. The composition according to any one of clauses 28 to 32. Clause 34. The composition of any one of clauses 28 to 33, having a three-dimensional structure that is a nanoworm or a nanorod. Clause 35. The composition of any one of clauses 28 to 34, wherein the nanorod has an aspect ratio of about 10:1 to about 1000:1. Clause 36. The composition of any one of clauses 28 to 35, which is a nanorod having a diameter of about 10 nm to about 20 nm and a length of about 1 micron to about 2 microns. Clause 37. The composition of any one of clauses 28 to 36, wherein the ratio of the compound represented by formula (I) to the compound represented by formula (IV) is from about 0.01:1 to about 1:0.01. EXAMPLES

[0090] experiment: measurement Nuclear Magnetic Resonance (NMR). All NMR spectra were recorded on a Bruker DRX 400 or 500 MHz spectrometer using an external lock (CDCl3 or DMSO-d6) and referenced to the remaining non-deuterated solvent (CHCl3 or DMSO).

[0091] Size Exclusion Chromatography (SEC) and Triple Detection-Size Exclusion Chromatography (TD-SEC). Analysis of the molecular weight distribution of the polymers was determined using a Polymer Laboratories GPC50 Plus equipped with a differential refractive index detector. The absolute molecular weight of the polymers was determined using a Polymer Laboratories GPC50 Plus equipped with a dual angle laser light scattering detector, a viscometer and a differential refractive index detector. High performance liquid chromatography (HPLC) grade N,N-dimethylacetamide (DMAc, containing 0.03 wt% LiCl) was used as the eluent at a flow rate of 1.0 mL / min. Separation was achieved using two PLGel Mixed B (7.8 × 300 mm) SEC columns connected in series and held at a constant temperature of 50 °C. The triple detection system was calibrated using a 5 mg / mL 110K polystyrene (PSTY) standard. Samples of known concentration were freshly prepared in DMAc + 0.03 wt% LiCl and passed through a 0.45 μm PTFE syringe filter before injection. Absolute molecular weights and dn / dc values ​​were determined based on quantitative mass recovery methods using Polymer Laboratories Multi Cirrus software.

[0092] Transmission Electron Microscopy (TEM). A JEOL-1010 transmission electron microscope was used to analyze the nanostructured appearance of the polymer lattices at ambient temperature using an accelerating voltage of 100 kV with a spot size of 5. A typical TEM grid was prepared as follows: the nanostructured sample was diluted to about 0.05 wt% with Milli-Q water at 34° C. Then, a formvar preheated (34° C.) copper TEM grid was immersed in the diluted solution and the excess solution was blotted with filter paper and then dried at 34° C.

[0093] Matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF). Mass spectrometry spectra were recorded using a Bruker autoflex III smartbeam operated in reflector mode. Ions were accelerated at a potential of 20 kV using a nitrogen laser emitting at 337 nm. The polymer solution concentrations were 1 mg / mL in tetrahydrofuran (THF), 1 mg / mL in THF for sodium trifluoroacetate (NaTFA) and 10 mg / ml for DCTB (T-2-(3-(4-t-butylphenyl)-2-methyl-2-propenylidene)malononitrile). To prepare the samples for measurement, 20 μL of polymer solution, 20 μL of DCTB solution and 2 μL of NaTFA solution were mixed in an Eppendorf tube, vortexed and centrifuged. 1 μL of solution was placed on a sample drop plate and dried at ambient conditions before proceeding with the measurement.

[0094] UV-Vis spectrometer. UV-Vis absorption spectra were recorded on a UV-Vis Cary 4000 spectrophotometer at 25 °C.

[0095] Confocal microscopy. Confocal microscopy images of fluorescent probe-labeled nanorods dispersed in Milli-Q water at 20 mg / mL. Each sample of 5SL was dropped onto a glass slide. Confocal microscopy images were taken using a confocal LSM Zeiss 710 laser scanning microscope (inverted) equipped with an oil immersion objective (1.40 Oil DIC M27 / 63x). Two excitation wavelengths of 488 nm and 561 nm were used for Oregon green 488 and SAv-DyLight 550, respectively.

[0096] Dynamic Light Scattering (DLS). Dynamic light scattering measurements were performed using a Malvern Zetasizer Nano Series 3000HS running DTS software and operating with a 4mW He-Ne laser at 633 nm. The analysis was performed at an angle of 173°. The sample refractive index (RI) was set to 1.59 for polystyrene. The viscosity of the dispersant and the RI of water were 0.4071 mP (70°C) and 1.33 Ns / m, respectively. 2The hydrodynamic number-average particle size and particle size distribution (PSD) were reported. The PSD was used to describe the width of the particle size distribution. It was calculated by cumulative analysis of the DLS measured intensity autocorrelation function and was related to the standard deviation of a hypothetical Gaussian distribution (i.e., PSDDLS=σ 2 / ZD 2 (where σ is the standard deviation and ZD is the Z-average mean size).

[0097] To determine the lower critical solution temperature (LCST) of all PNIPAM macro-CTAs, the polymer macro-CTAs were dissolved in Milli-Q water in an ice bath at a concentration of 5 mg / mL or 53.8 mg / mL, and SDS was added at a concentration of 0.21 mg / mL or 2.23 mg / mL, respectively. The solutions were then filtered directly into DLS cuvettes using a 0.45 μm cellulose syringe filter. The polymer solutions were cooled to 5 °C, and the cuvettes were placed in the DLS spectrometer. Measurements were performed by slowly increasing the temperature from 5 °C to 70 °C with a ramp rate controlled by the SOP (Standard Operating Procedure) software.

[0098] material: Synthesis of functional polymer nanostructured materials All chemicals were used as received unless otherwise stated. Solvents used were either HPLC or AR grade solvents and included dichloromethane (DCM; Aldrich AR grade), dimethylformamide (DMF; Aldrich, AR grade) and tetrahydrofuran (THF; Labscan, HPLC grade), DMSO (Aldrich, 99.9%). Basic activated alumina (Aldrich: Brockmann I, standard grade, ~150 mesh, 58A), Milli-Q water (Biolab, 18.2 M=m), sodium dodecyl sulfate (SDS; Aldrich, 99%), triethylamine (TEA; Fluka, 99%), N,N'-dicyclohexylcarbodiimide (DCC, Aldrich, 99%), N-hydroxysuccinimide (NHS, Aldrich, 98%), ethanolamine (Aldrich, >99%), 4-(dimethylamino)pyridine (DMAP, Merck, 99%), copper(II) sulfate (Aldrich, 99%), L-ascorbic acid (Aldrich, 99%), 1-butanethiol (Aldrich, 99%), propargyl alcohol (Aldrich, 99%), sodium azide (Aldrich, 99.5%), methacryloyl chloride (Fluka, 97%), 3-chloro-1-propanol (Aldrich, 98%), poly(ethylene glycol) monomethyoxyether-2000 (mPEG, Aldrich), p-toluenesulfonyl chloride (Aldrich, >99%), β-cyclodextrin (Sigma, >97%), allylamine (Aldrich, 98%), Aldrithiol™-2 (DPDS, Aldrich, 98%), 2-mer Captoethanol (Merck, >98%), dopamine hydrochloride (Sigma), DL-homocysteine ​​thiolactone hydrochloride (Tla, Aldrich, >99%), biotin (Sigmal-Aldrich, >99%), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC·HCl, Fluka, >98%), reduced L-glutathione (Sigma-Aldrich, >98%), Streptomyces avidinii (Streptomyces Streptavidin from S. avidinii (SAv, Sigma), streptavidin DyLight 550 conjugate (Pierce), Oregon Green® 488 maleimide (Molecular Probes®), carbon disulfide (Aldrich, >99.9%), 2-bromo-2-methylpropionic acid (Aldrich, 98%) and methyl-2-bromopropionate (MBP; Aldrich, 98%) were used as received. Styrene (STY: Aldrich, >99%) was passed through a basic alumina column to remove inhibitors. N-isopropylacrylamide (NIPAM: Aldrich, 97%) was recrystallized from (n-hexane / toluene, 9 / 1, v / v) and azobisisobutyronitrile (AIBN, Riedel-de Haen) was recrystallized twice from methanol before use.

[0099] The RAFT agent methyl 2-(butylthiocarbonothioylthio)propanoate (MCEBTTC) was synthesized according to a previous procedure. (CN Urbani, MJ Monteiro, Macromolecules, 2009, 42, 3884-3886.)

[0100] Synthesis of Carboxylic Acid Functional RAFT Agents (Acid-RAFT) Potassium phosphate tribasic (16.6 g, 0.0782 mol) was dissolved in 130 mL of acetone in a 500 mL round bottom flask. The mixture was kept stirring for 5 hours to prepare a pale yellow suspension. 1-Butanethiol (8.0 mL, 0.0742 mol) was added to the mixture and stirred for 1 hour. Carbon disulfide (9.1 mL, 0.151 mol) was then added dropwise to the stirred mixture and stirred at 0.5°C (in ice bath) for 2 hours. 2-Bromo-2-methylpropionic acid (11.7 g, 0.07 mol) was added to the mixture under stirring and the mixture was allowed to react at room temperature (23°C) overnight. The solid was isolated by filtration and the solvent volume was reduced by using rotary evaporation. The residue was then diluted with cold 10% HCl solution (4 x 50 mL) and stirred at room temperature overnight. The solution was then extracted twice with n-hexane, dried over anhydrous MgSO4, filtered, and the solvent was removed by rotary evaporation. The residual yellow solid was purified by column chromatography (on silica, petroleum spirit / ethyl acetate: 3 / 2, Rf=0.51). The solvent was then removed by rotary evaporation and the compound was dissolved in n-hexane and stored in a freezer to crystallize. The product was filtered and dried under high vacuum for 24 hours, giving a yield of 61%. 1 H NMR (CDCl3, 298K, 300MHz):ppm 3.27(t,2H;J=7.38Hz;CH3CH2CH2CH2S-), 1.70(s,6H;(CH3)2-), 1.64(m,2H,J=8.94Hz;CH3CH2CH 2CH2S-), 1.41(m,2H;J=7.29Hz;CH3CH2CH2CH2S-), 0.90(t,3H,J=7.26Hz;CH3CH2CH2CH2S-);13C NMR (CDCl3, 298K, 75MHz): 228.3, 128.9, 55.6, 36.7, 29.8, 25.2, 22.1, 13.6.

[0101] Synthesis of pyridyl disulfide-functional RAFT agents (PDS-RAFT) The reaction mixture was allowed to warm to room temperature and stirred for 24 hours. DCM was removed by rotary evaporation. The residue was redispersed in diethyl ether and filtered. The filtrate was concentrated to a viscous residue and purified by silica column chromatography (1 / 4 petroleum spirit / ethyl acetate) (yield: 59.4%). Scheme 2 [ka]

[0102] Synthesis of dopamine-functional RAFT agents (Dopa-RAFT) As shown in Scheme 3, RAFT-acid (3.0 g, 0.012 mol), NHS (1.5 g, 0.013 mol) and DCC (2.67 g, 0.013 mol) were dissolved in 50 mL of DCM, which was cooled to 0° C. in an ice bath. The reaction mixture was allowed to warm to room temperature and stirred for 2 h. The reaction was monitored by TLC for complete conversion to the RAFT-active ester. The mixture was then filtered into another round-bottom flask equipped with a stirrer. Dopamine hydrochloride (2.0 g, 0.011 mol) and TEA (1.7 mL, 0.012 mol) were dissolved in 10 mL of DMF and added dropwise to the above RAFT-active ester. The reaction mixture was stirred for another 24 h. The color of the reaction mixture turned brown. 50 mL of DCM was added to the reaction mixture. The mixture was then washed with 0.1 N HCl solution followed by Milli-Q water. The color of the DCM phase turned yellow. The DCM phase was dried over MgSO4, filtered and concentrated. The filtrate was concentrated to a viscous residue and purified by silica column chromatography (1 / 1 petroleum spirit / ethyl acetate) (yield: 44.3%). Scheme 3 [ka]

[0103] Synthesis of biotin-functional RAFT agents (biotin-RAFT) Synthesis of Hydroxyl-Functional RAFT (RAFT-OH) As shown in Scheme 4, RAFT-acid (1.75 g, 6.9 × 10 -3mol), NHS (0.95 g, 8.3 × 10 -3 mol) and DCC (1.72 g, 8.3 × 10 -3 10 mol) was dissolved in 30 mL of DCM and cooled to 0° C. in an ice bath. The reaction mixture was allowed to warm to room temperature and stirred for 2 h. The reaction was monitored by TLC for complete conversion to the RAFT-active ester. The mixture was then filtered into another round-bottom flask equipped with a mechanical stirrer. Ethanolamine (0.4 g, 6.5×10 -3 mol) was dissolved in 10 mL of DMF and added dropwise to the above RAFT-activated ester. The reaction mixture was stirred for an additional 24 h. The mixture was filtered, concentrated to a viscous residue and purified by silica column chromatography (1 / 2 petroleum spirit / ethyl acetate) (yield: 55.0%). Scheme 4 [ka] Synthesis of biotin-functionalized RAFT (biotin-RAFT)

[0104] As shown in Scheme 5, RAFT-OH (1.10 g, 3.73 × 10 -3 mol), biotin (0.91 g, 3.73 × 10 -3 mol), EDC.HCl (1.42g, 7.45×10 -3 mol) and DMAP (45.5 mg, 3.73 × 10 -3 (mol) was dissolved in 20 mL of DMF and cooled to 0° C. in an ice bath. The reaction mixture was allowed to warm to room temperature and stirred for 48 h. The reaction mixture was then diluted with 100 mL of DCM and washed five times with deionized water. The DCM phase was then dried over MgSO4, filtered, and concentrated to a viscous residue and purified by silica column chromatography (1 / 8 methanol / DCM) (yield: 69.6%). Scheme 5 [ka] Synthesis of functional poly(NIPAM) macro-CTAs

[0105] All functional poly(NIPAM) macro-CTAs were synthesized by reversible addition-fragmentation chain transfer polymerization (RAFT polymerization) in DMSO at 60 °C. The feed ratio of NIPAM / RAFT agent / AIBN was kept at 44 / 1 / 0.1 for all different functional RAFT agents. The ratio of DMSO to NIPAM was kept at 2 / 1 (v / w). Typically, (10 g, 8.85 × 10 -2 mol), methyl-RAFT (0.58 g, 2.0 × 10 -3 mol) and AIBN (3.3 mg, 2.0 × 10 -4 mol) was dissolved in 20 mL of DMSO. The mixture was purged with argon for 30 min and then heated at 60° C. for 16 h. The reaction was stopped by cooling to 0° C. in an ice bath and exposed to air. The solution was then diluted with 500 mL of DCM and washed with Milli-Q water (5×100 mL). The DCM phase was then dried over anhydrous MgSO4, filtered, and concentrated by rotary evaporation. The polymer was recovered by precipitation in a large excess of diethyl ether (500 mL), isolated by filtration, and then dried under vacuum at room temperature for 24 h to give a yellow powder product (45% yield). 1 The conversion was 96% as determined by 1 H NMR spectroscopy.

[0106] Table 1 summarizes the data for the functional poly(NIPAM) macro-CTAs.

[0107] [Table 1]

[0108] RAFT-Mediated Emulsion Polymerization of Styrene with Functional Poly(NIPAM) Macro-CTA in Water The polymerization was as follows: The cumulative mass of poly(NIPAM) macro-CTA was (0.35 g). Other components present in the reaction mixture were styrene (0.35 g); SDS (14.5 mg), AIBN (1.2 mg), Milli-Q water (6.25 g). Dopa-poly(NIPAM) macro-CTA 3 (0.105 g, 1.33 × 10 -5mol), biotin-poly(NIPAM) macroCTA 4 (0.07 g, 1.66 × 10 -5 mol) and SDS (7.25 mg, 5 × 10 -5 AIBN (0.6 mg, 3.7 × 10 mol) was dissolved in cold water (3.125 g) in a Schlenk tube. The mixture was deoxygenated by purging with argon for 15 min. -6 mol) to styrene (0.175 g, 1.7 × 10 -3 ) and the solution was then injected into a Schlenk flask. The mixture was then purged with argon in an ice bath for an additional 10 minutes and then heated to 70°C for 3.5 hours. The reaction was quenched by exposure to air at 70°C. The polymer emulsion was then analyzed by SEC, 1 It was characterized by H NMR and DLS, and the data are shown in Table 2.

[0109] [Table 2]

[0110] Furthermore, transmission electron microscopy images of the 60% Dopa-functional / 40% biotin-functional composition show that the three-dimensional structure of the composition is a nanoworm. 1 H NMR showed an average of 45 repeating NIPAM units and 29-30 repeating PSTY units in each of the diblock polymers.

[0111] Coating surfaces with nanoworms The glass slips (and silicon wafers) were washed sequentially with aqueous 10% HCl, acetone and methanol. The slips were then dried under a stream of nitrogen. In four 50 ml plastic vials (a, b, c and d), 10 mg of 60% Dopa- and 40% biotin-functionalized nanoworms were added in each vial, dispersed in 4 ml of tris(hydroxymethyl)aminomethane (Tris) solution (10 mM, pH=8.5) and allowed to rehydrate for 20 min. The glass slips and silicon wafers were added. Different amounts (1, 60, 120, 240 μL) of dopamine solution (33 mg / ml) were then added to vials a, b, c and d, respectively. The mixture was then shaken for 4 h. The silicon wafers were removed, washed with water and dried for SEM imaging. The glass slips were washed with water and then immersed in 0.5 ml of SAv-DyLight 550 protein solution for 1 min. The slips were then washed again with water and stored in water for fluorescence microscopy.

[0112] Fluorescence images (a–d) of 60% Dopa- and 40% biotin-functionalized nanoworms on glass slips to which different amounts of free dopamine ((a) 0 mg / ml, (b) 0.5 mg / ml, (a) 1.0 mg / ml, and (e) 2.0 mg / ml) had been added and then treated with SAv-550 solution showed that biotin incorporation was nearly quantitative on the surface of the worms.

[0113] SEM (e–h) of silicon wafers coated with 60% Dopa- and 40% biotin-functionalized nanoworms showed that 4 h of coating and 1.0 mg / ml of free dopamine gave the best coating on both the glass slip and silicon wafer. Synthesis of maleimide-functional cationic polymers Synthesis of furan-protected maleimide initiators [ka] The initiator was synthesized according to the literature (Geng, et al., J. Am. Chem. Soc., 2007, 129(49), pp 15156-15163).

[0114] Synthesis of furan-protected maleimide polydimethylamino EA (polyDMAEA) DMAEA 15ml (0.1mol), initiator 0.197g (6.6×10 -4 mol), CuCl2 8.8 mg (6.6 × 10 -5 mol), Tris[2-(dimethylamino)ethyl]amine (Me6TREN) 61 mg (2.6 × 10 -4 A solution of 1.2 mmol of 1,2-dichlorophenyl ether was added to 7.5 ml of isopropanol and purged with argon for 30 min. 12.6 mg of Cu(0) (<425 μm) was added under argon. Polymerization was allowed to proceed at room temperature for 6.5 h, with a conversion of 41%. The polymerization mixture was then diluted with acetone and passed through neutral Al2O3 to remove copper, and the solution was concentrated, precipitated three times in petroleum spirits, and dried under high vacuum, yielding 3.5 g of polymer. M by NMR was n = 9740. Figure 1 shows the structure of the polymer in deuterated chloroform (CDCl3). 1 1 H nuclear magnetic spectrum. For the polymer shown in FIG. 1, n is an integer equal to 61.

[0115] Synthesis of the quaternized furan-protected maleimide PDMAEA Poly-DMAEA was quaternized using methyl iodide and 1-iodoalkanes of different chain lengths (as shown in Table 3) as shown in Scheme 6. Poly-DMAEA was reacted with long chain iodoalkanes at 60° C. for 8 hours. The solution was cooled to room temperature and the corresponding amount of iodomethane was added. The reaction was allowed to stir at room temperature overnight. The reaction mixture was then precipitated in acetone to give a pale yellow powder, which was then dried under high vacuum. Scheme 6 [ka]

[0116] [Table 3]

[0117] Deprotection of the quaternized furan-protected maleimide polyDMAEA As shown in Scheme 7, the quaternized polyDMAEA was then dissolved in DMSO (0.2 g in 4 ml) and then heated at 120° C. for 3 h. The residue was then precipitated in acetone to give a light brown polymeric material. The polymer was characterized by NMR to confirm the maleimide chain ends. The percentage of long carbon chains was very close to the target amount (~10%). Scheme 7 [ka] Figures 2A, 2B, and 2C show the results of the reaction of maleimide-functionalized quaternized polyDMAEA in DO. 1 H NMR spectra are shown (n=61): (Figure 2A) 10 mol% 1-iodooctane, (Figure 2B) 10 mol% 1-iodododecane, and (Figure 2C) 10 mol% 1-iodohexadecane.

[0118] Conjugation of maleimide-functional cationic polymers with Dopa- and PDS-functional nanoworms Figure 3 is a scheme showing the conjugation of maleimide-functional cationic polymers with Dopa- and PDS-functional nanoworms. Maleimide-functional cationic polymers A, B or C (12.7 mg each) were placed in an Eppendorf tube and 1 ml of water was added. After the polymers were dissolved, a solution of polymer A, B or C was added to another Eppendorf tube containing nanoworms Dopa-(60%) and PDS-(40%) (20 mg nanoworms). Then, 10 μl of TCEP solution (32.5 mg / ml) was added, the Eppendorf tube was wrapped in aluminum foil (protected from light) and the tube was shaken overnight. The final products had the chemical structures shown below, where nanoworm A had n=7, nanoworm B had n=11 and nanoworm C had n=15. [ka]

[0119] Antibacterial Testing Antibacterial testing of maleimide-functional cationic polymers A, B and C in solution Experimental Setup For the maleimide-functional cationic polymers A, B and C, 5 mg / ml solutions were prepared in water. · To a 24-well plate, 500 μl of polymer solution in water was added in duplicate and 500 μl of water was added to the LB blank and E. coli containing wells. 1x10 E. coli solution in the well containing the polymer in water 5 500 μl of cells / ml was added. 500 μl of LB was added to blank wells. The top of the plate was covered with Parafilm, then the lid was placed on top, and then more Parafilm was used to seal the edges to prevent solvent evaporation. The plates were then placed in a 37°C incubator and shaken at 120 rpm for 14 hours.

[0120] The antibacterial activity of the polymers was measured based on the optical density of the E. coli culture. Figure 4 is a bar graph showing the inhibitory effect of the polymers on the E. coli culture. All three polymers showed significant inhibition (>90%) of bacterial growth.

[0121] Antibacterial testing of functional cationic nanoworms A, B and C in solution Experimental Setup · For nanoworms A, B and C, 5mg / ml solutions were prepared in water. · To a 24-well plate, 500μl of the nanoworm solution in water was added in duplicate, and 500μl of water was added to the LB blank and wells that would receive E. coli. 1x10 E. coli solution in the well containing the polymer in water 5 500 μl of cells / ml was added. 500 μl of LB was added to blank wells. The top of the plate was covered with Parafilm, then the lid was placed on top, and then more Parafilm was used to seal the edges to prevent evaporation. The plate was placed in a 37°C incubator and shaken at 120 rpm for 14 hours. · Because of the color of the solution, E. coli could not be measured using the usual method of measuring OD. Growth of E. coli was monitored by plating the nanoworm / E. coli solution on LB agar plates. Dilutions were made in LB and 20 μl were plated and left in the incubator turned on.

[0122] Figures 5A-5D are scanning electron microscopy images showing the antibacterial activity of cationic polymer nanoworms A, B, and C against E. coli. (Figure 5A) LB blank and E. coli controls, (Figure 5B) nanoworm B, (Figure 5C) nanoworm C, and (Figure 5D) nanoworm D, all at different dilutions (10 as indicated). 2 , 10 4 and 10 6 )). All nanoworms could kill almost 100% of the bacteria. Only one E. coli colony was found (circled in Figure 5D). Meanwhile, many E. coli colonies were found on the +ve control plate (Figure 5A). This indicated that the cationic polymer nanoworms could efficiently kill almost all E. coli.

[0123] Overall, it was revealed that the use of SAv-biotin binding experiments and SEM demonstrated that nanoworms can be effectively coated onto glass slips and silicon wafers. Maleimide-functionalized cationic polymers showed significant inhibition (>90%) of E. coli growth. Further conjugation of these maleimide-functionalized nanoworms onto PDS- and Dopa-functionalized nanoworms retained the antibacterial properties.

[0124] Overall, the compounds and compositions of the present disclosure can provide antimicrobial nanostructures that allow targeted binding and cytopathogenesis of microorganisms. The ratiometric control of the functionality of the compositions of the present disclosure provides the ability to target and modify a wide range of microorganisms, from bacteria to viruses.

[0125] definition The term "pharmaceutical acceptable" means suitable for use in pharmaceutical preparations, generally regarded as safe for such use, officially approved for such use by a regulatory agency of a national or state government, or listed in the US Pharmacopoeia or other generally recognized pharmacopoeias for use in animals, and especially in humans.

[0126] The term "pharmaceutically acceptable salt" refers to a salt that can improve the desired pharmacological activity. Examples of pharmaceutically acceptable salts include acid addition salts formed with inorganic or organic acids, metal salts, and amine salts. Examples of acid addition salts formed with inorganic acids include salts with hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Examples of acid addition salts formed 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, p-chlorobenzenesulfonic acid, and the like. Examples of acid addition salts formed with acids, 2-naphthalenesulfonic acid, p-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]oct-2-enyl-carboxylic acid, glucoheptonic acid, 4,4'-methylenebis(3-hydroxy-2-naphthoic acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid and muconic acid. Examples of metal salts include salts with sodium, potassium, calcium, magnesium, aluminum, iron and zinc ions. Examples of amine salts include salts with ammonia and organic nitrogenous bases strong enough to form salts with carboxylic acids.

[0127] The term "therapeutically effective amount" refers to the amount of a compound that, when administered to a subject for treating a condition, is sufficient to treat the condition. A "therapeutically effective amount" can vary depending on the compound, the condition and its severity, the age and weight of the subject being treated.

[0128] The term "virus" refers to submicroscopic infectious agents that comprise complex non-living molecules that typically contain a proteinaceous coat surrounding an RNA or DNA core of genetic material but no semipermeable membrane, are capable of growing and multiplying within living cells, and can cause disease in humans, animals or plants.

[0129] The compounds of the present disclosure include tautomers, geometric isomers or stereoisomers of the compounds.The ester, oxime, onium, hydrate, solvate and N-oxide forms of the compounds are also encompassed by the present disclosure.The present disclosure considers that all such compounds are encompassed by the present disclosure, including cis- and trans-geometric isomers (Z- and E-geometric isomers), R- and S-enantiomers, diastereomers, d-isomers, l-isomers, atropisomers, epimers, conformers, rotamers, mixtures of isomers and their racemates.

[0130] The description of various aspects of the present disclosure is presented for illustrative purposes and is not intended to be exhaustive or to be limited to the disclosed aspects. Many modifications and variations will become apparent to those skilled in the art without departing from the scope and spirit of the described aspects. The terminology used in this specification is selected to best explain the principles, practical applications or technical improvements of the aspects compared to the technology found in the market, or to allow other skilled in the art to understand the aspects disclosed herein.

Claims

[Claim 1] Polystyrene block; Poly(N-isopropylacrylamide) blocks; formula: 【Chemistry 1】 (In the formula, n is an integer from 10 to 100, Each z is independently an integer from 1 to 20. a plurality of ammonium moieties represented by formula: 【Chemistry 2】 (In the formula, Q is selected from the group consisting of fluoro, chloro, bromo, and iodo; each m is an integer from 1 to 20; Each q and each x is independently an integer from 1 to 20. a plurality of ammonium moieties represented by formula: 【Chemistry 3】 , optionally of the formula: 【Chemistry 4】 wherein v is an integer from 1 to 20; and 【Chemistry 5】 wherein j and p are independently an integer from 1 to 20, or a pharma- ceutically acceptable salt thereof; a second polystyrene block; A second poly(N-isopropylacrylamide) block; formula: 【Chemistry 6】 , optionally of the formula: 【Chemistry 7】 wherein v is an integer from 1 to 20; and formula: 【Chemistry 8】 (In the formula, R １ is selected from the group consisting of -O- and -NH-; R ２ Ha-CH ３ , biotin, pyridyl disulfide, 【Chemistry 9】 a second compound comprising a moiety represented by the formula: A coating composition for depositing on at least a portion of an object, the coating composition comprising a nanorod or nanoworm comprising:

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