Compounds for the treatment of diseases

Abstract

Disclosed herein are compounds for inhibiting the kinase activities of cytidine / uridine monophosphate kinase 2 (CMPK2), which is essential for the activation of the Nod-like receptor protein 3 (NLRP3) inflammasome. Also disclosed are methods of treating diseases associated with Aberrant NLRP3 inflammasome activation.

Description

Compounds for the Treatment of DiseasesCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to Provisional Application No. 63 / 662,654, filed June 21, 2024, the disclosures of which is hereby incorporated by reference in the entirety.TECHNICAL FIELD

[0002] The patent document relates to compounds for inhibiting the kinase activities of cytidine / uridine monophosphate kinase 2 (CMPK2), which is essential for the activation of the Nod-like receptor protein 3 (NLRP3) inflammasome. Also provided herein are methods of treating diseases associated with Aberrant NLRP3 inflammasome activation.BACKGROUND

[0003] Previous work has shown that the activity of the CMPK2 thymidylate kinase domain is essential for activation of the NLRP3 inflammasome, a multiprotein complex necessary for the maturation of pro- inflammatory cytokines like IL-lbeta (IL-ip) and IL-18. Aberrant NLRP3 inflammasome activation is a major driver of chronic inflammatory conditions, and has potential roles in cancer, metabolic diseases, neuroinflammatory diseases, diabetes and chronic pain.

[0004] Current common anti-inflammatory medications, NSAIDs and corticosteroids, have numerous side effects and contraindications due to the fact that these medications broadly target inflammatory pathways. A need exists to develop new compounds and therapeutic approaches for treating diseases associated with Aberrant NLRP3 inflammasome activation.SUMMARY

[0005] This patent document discloses compounds specifically targeting CMPK2-dependent activation of the NLRP3 inflammasome. This strategy represents a new paradigm for the control and treatment of a broad range of inflammatory processes and diseases. In particular, these compounds provide a viable alternative for patients with contraindication for NSAIDs and corticosteroids.

[0006] An aspect of the patent document provides a compound of Formula I or a pharmaceutically acceptable salt thereof. The structure of Formula I is as follows.Formula IWherein:M in each instance is independently O or S, m is 0, 1, or 2, n is 1, 2 or 3,X is selected from the group consisting of O, CH2, NH, CHF, CF2, CBr2, CCI2, and S,Y is selected from the group consisting of H, C1-12alkyl-C(O), C1-12alkyl-C(O)-O-C1-8alkylene-C(O), phenyl, phenyl-C(O), 5-6-membered heteroaryl-C(O), C1-6alkyl-O-C(O), C1-6alkyl-C(O)-O-C1- ealkylene, C1-6alkyl-O-C(O)-O-C1-6alkylene, P(O)(Ra)2, and P(S)(Ra)2, Rb-CH(N(RC)2)C(O),Wherein each Rais independently selected from the group consisting of OH, Ci-nalkyl- C(O)-O-C1-8alkylene-O, C1-6alkyl-O-C(O)-O-C1-4alkylene-O, Ci-2oalkyl-0-C1-8alkylene-0, Ci-2oalkyl-S-C1-8alkylene-0, C1-12alkyl-C(O)C1-8alkylene-O, phenyl-O, naphthyl-O, 5-12- membered heteroaryl-O, C1-6alkyl-phenyl-O, C1-6alkyl-O-C(O)-C1-4alkylene-NH, C1- 6alkyl-O-C(O)-CH(Rm)-NH, phenyl-O-C(O)-CH(Rm)-NH, heteroaryl-O-C(O)-CH(Rm)- NH, C1-8alkyl-C(O)-S-C1-8alkylene-O, C1-6alkyl-O-C(O)-S-C1-4alkylene-O, Ci-2oalkyl-S- C1-8alkylene-O, C1-6alkyl-S-C(O)-C1-4alkylene-NH, R°-C(O)-O-Ph-C1-4alkylene-O, C1- i8alkyl-O-CH2CH(ORp)CH2, Ci-i8alkyl-S-CH2CH(ORp)CH2, wherein the phenyl or naphthyl in Rais optionally substituted with one or more of halogen, C1-6alkyl, C3- ecycloalkyl, haloC1-6alkyl, CN, and OC1-6alkyl, wherein each Rmis independently a side chain (e.g., methyl, isopropyl, phenyl, etc.) of an amino acid, wherein each Rnis independently H or C1-6alkyl, wherein each R° is independently Ci-i8alkyl, C3-6cycloalkyl, Ci-salkylene-C3-6cycloalkyl, phenyl, 5-12- membered heteroaryl or PEG (50-500 Da), wherein each Rpis independently H, Ci-i8alkyl, C1-18alkyl-C(O), benzyl, or C2-i8alkenyl, wherein Rbis a side chain of an amino acid, wherein each Rcis independently H or C1- ealkyl,Wherein the alkyl or alkylene of Y (e.g. C1-8alkyl of C1-8alkyl-C(O)) is optionally substituted with one or more of halogen, CN, OH, SH, (e.g. side chain of an amino acid), 5-10 membered heteroaryl, and phenyl, N(R°)2, wherein each R° is independently H or C1- salkyl, wherein the phenyl and heteroaryl in Y are each optionally substituted with C1-4alkyl, CN, OH, halogen, and C1-4alkyl-O, one of R1and R2is H and the other is selected from the group consisting of H, OH, halogen, CN, OC1- ealkyl, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl,R3is selected from the group consisting of H, halogen, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl, R4and R5are each independently H or C1-4alkyl,R6is selected from the group consisting of H, CN, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl, A is selected from the group consisting of O, S, CH2, CF2, CCI2, CH(OH), and CHF;B is selected from the group consisting ofWherein Rd, and Reare each independently selected from the group consisting of H, C1-6alkyl, OH, OC1-6alkyl, C3-6cycloalkyl, Ci-salkylene-C3-6cycloalkyl, OC3-6cycloalkyl, phenyl, Ci-3alkylene-phenyl, NH, and NC1-6alkyl,Rfand Rgare each independently selected from H, C1-6alkyl, halo- C1-6alkyl, Ci-salkylene-OH, C1- 3alkylene-OC1-6alkyl, C3-6cycloalkyl, Ci-3alkylene-C3-6cycloalkyl, CN, halogen,R1is selected from H, C1-6alkyl, C3-6cycloalkyl, andRhis selected from the group consisting of H, C1-6alkyl, C3-6cycloalkyl, OH, OC1-6alkyl, NH, and NC1- ealkyl,E is selected from the group consisting of O, S, CH2, CHF, CF2, NH, CBr2, CR>Rk, and CCI2,R> andRkare independently selected from the group consisting of H, C1-6alkyl, halogen; alternativelyRj and Rklink up to form a 2-6 membered cycloalkyl ring.

[0007] Another aspect of this patent document discloses a pharmaceutical composition comprising the compound described herein or the pharmaceutically acceptable salt, isomer, or prodrug thereof.

[0008] Another aspect of this disclosure provides a method of treating a disease in a subject comprising administering to the subject in need a therapeutically effective amount of a compound of Formula I, a pharmaceutically acceptable salt or isomer thereof, or a pharmaceutical composition thereof.DESCRIPTION OF THE DRA5WINGS

[0009] Figure 1 illustrates the synthesis of a compound of Formula I.

[0010] Figure 2 illustrates the synthesis of an intermediate.

[0011] Figure 3 illustrates the synthesis of a compound of Formula I.

[0012] Figure 4 illustrates the synthesis of a compound of Formula I.

[0013] Figure 5 illustrates the synthesis of a compound of Formula I.DETAILED DESCRIPTION

[0014] Various embodiments of this patent document provide compounds as inhibitors of CMPK2, which directly impact the activation of the NLRP3 inflammasome. Also provided are methods of treating diseases associated with aberrant activation of NLRP3 inflammasome. Different from conventional therapeutic efforts targeting directly theNLRP3 inflammasome, this strategy addresses the upstream regulatory processes (i.e., CMPK2).

[0015] The kinase activity of CMPK2 is essential for the activation of the Nod-like receptor protein 3 (NLRP3) inflammasome, the multiprotein complex responsible for the maturation and secretion pro-inflammatory cytokines IL-lbeta (IL-ip) and IL-18. It has been unambiguously demonstrated that CMPK2 is a pyrimidine nucleoside diphosphate kinase. The membrane-permeable compounds or ribonucleotide 3'-deoxy-3',4'-didehydrocytidine triphosphate (ddhCTP) prodrugs disclosed herein, including the ddhC nucleoside, can be converted to ddhCTP within mammalian cells and inhibit maturation and secretion of IL-ip from "inflamed" macrophages. The pharmacological blockade of CMPK2-dependent NLRP3 inflammasome activation represents a new paradigm for the control and treatment of a broad range of inflammatory processes and diseases. This strategy is fully differentiated from all current therapeutic efforts, which directly target the NLRP3 inflammasome and not the upstream regulatory processes (i.e., CMPK2).

[0016] While the following text may reference or exemplify specific embodiments of a compound or a method of treating a disease or condition, it is not intended to limit the scope of the compound or method to such particular reference or examples. Various modifications may be made by those skilled in the art, in view of practical and economic considerations, such as the substitutions of the compound and the amount or administration of the compound for treating or preventing a disease or condition.

[0017] The articles "a" and "an" as used herein refers to "one or more" or "at least one," unless otherwise indicated. That is, reference to any element or component of an embodiment by the indefinite article "a" or "an" does not exclude the possibility that more than one element or component is present.

[0018] The term “pharmaceutical composition” refers to a mixture of a compound disclosed herein with other chemical components, such as diluents or additional carriers. The pharmaceutical composition facilitates administration of the compound to an organism. Multiple techniques of administering a pharmaceutical composition exist in the art including, but not limited to, oral, injection, aerosol, parenteral, and topical administration. In some embodiments, pharmaceutically acceptable salts of the compounds disclosed herein are provided.

[0019] The term "subject" encompasses any animal, but preferably a mammal, e.g., human, non- human primate, a dog, a cat, a horse, a cow, or a rodent. More preferably, the subject is a human.

[0020] The term “carrier” refers to a chemical compound that facilitates the incorporation of a compound into cells or tissues.

[0021] The term “diluent” refers to chemical compounds diluted in water that will dissolve the composition of interest as well as stabilize the biologically active form of the compound. Salts dissolved in buffered solutions are utilized as diluents in the art. One commonly used buffered solution is phosphate buffered saline because it mimics the salt conditions of human blood. Since buffer salts can control the pH of a solution at low concentrations, a buffered diluent rarely modifies the biological activity of a compound. As used herein, an “excipient” refers to an inert substance that is added to a composition to provide, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegrating ability, etc., to the composition. A “diluent” is a type of excipient.

[0022] The term “physiologically acceptable” or “pharmaceutically acceptable” refers to a carrier or diluent that does not abrogate the biological activity and properties of the compound.

[0023] The term “therapeutically effective amount” refers to an amount of a compound effective to prevent, alleviate or ameliorate symptoms of disease or prolong the survival of the subject being treated. Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.

[0024] The term "alkyl" refers to monovalent saturated alkane radical groups particularly having up to about 18 carbon atoms, more particularly as a lower alkyl, from 1 to 8 carbon atoms and still more particularly, from 1 to 6 carbon atoms. The hydrocarbon chain may be either straight-chained or branched. The term "Cnio alkyl" or "C1-C10 alkyl" refers to alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Similarly, the term " C1-4alkyl" refers to alkyl groups having 1, 2, 3, or 4 carbonatoms. Non-limiting examples of alkyls include groups such as methyl, ethyl, n-propyl, isopropyl, n- butyl, iso-butyl, tert-butyl, n-hexyl, n-octyl, tert-octyl and the like.

[0025] The term "alkylene" refers to a divalent hydrocarbon which may be either straight- chained or branched. Different from alkyl which has only one point of bonding with other groups or atoms, alkylene has two points of bonding. Non-limiting examples include groups such as CH2, (CFFX CH2CH(CH3), and the like. A C1-6 alkylene has 1, 2, 3, 4, 5 or 6 carbons. A C1-4 alkylene has 1, 2, 3 or 4 carbons.

[0026] The term “C1-4 alkoxy” includes an alkyoxy group having 1, 2, 3 or 4 carbons.

[0027] The term “carbocycle” or "cycloalkyl" refers to 3 to 10 membered cyclic hydrocarbyl groups having only carbon atoms as ring atoms and having a single cyclic ring or multiple condensed rings, including fused and bridged ring systems, which optionally can be substituted with from 1 to 3 alkyl groups. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, 1 -methylcyclopropyl, 2-methylcyclopentyl, 2- methylcyclooctyl, and the like, and multiple ring structures such as adamantanyl, and the like.

[0028] The term “haloalkyl” refers to a Ce-ioalkyl chain, straight or branched, in which one or more hydrogen has been replaced by a halogen. Non-limiting examples of haloalkyls include CHF2, CFH2, CF3, CH2CF2, CH2CF3, and CH2CH2F. In some embodiments, the alkyl in haloalkyl has 1, 2, 3 or 4 carbons.

[0029] The term “heterocycle” or "heterocycloalkyl" refers to 3 to 10 membered substituted or nonsubstituted non-aromatic cyclic groups where one or more carbon ring atoms are replaced with hetero atoms or groups containing heteroatoms (e.g. NH, NCl-C4alkyl O, and S). Nonlimnting examples include pyrrolidine, piperidine, N-methyl-piperizine, and morpholine. Optional substituents include C1-6 alkyl, C1-4 alkoxy, halogen, haloalkyl, sulfonamido, and amido.

[0030] The term “aryl” is intended to mean any stable monocyclic or bicyclic carbon ring of up to 7 members in each ring, wherein at least one ring is aromatic and all ring atoms of the aromatic ring are carbon atoms. Typical aryl groups include, but are not limited to, groups derived from aceanthrylene, acephenanthrylene, anthracene, azulene, benzene, fluoranthene, fluorene, hexacene, hexaphene, hexylene, as-indacene, s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, and the like. Particularly, an aryl group comprises from 6 to 10 or 6 to 14 carbon atoms.

[0031] The term "hetero" when used to describe a compound or a group present on a compound means that one or more carbon atoms in the compound or group have been replaced by a nitrogen,oxygen, or sulfur heteroatom. Hetero may be applied to any of the hydrocarbyl groups described above such as alkyl, e.g. heteroalkyl, cycloheteroalkyl.

[0032] The term “heteroaryl” refers to groups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms, having 6, 10, or 14 7t electrons shared in a cyclic array, wherein at least one ring atom contributing to the shared TI electrons in the cyclic array is a heteroatom. Typical heteroaryl groups include, but are not limited to, groups derived from acridine, carbazole, cinnoline, furan, imidazole, indazole, indole, indoline, indolizine, isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, phenanthridine, phenanthroline, phenazine, phthalazine, phthalimide, pteridine, purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine, quinazoline, quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene, and the like. Preferably, the heteroaryl group is between 5-15 membered heteroaryl, with 5-10 membered heteroaryl being particularly preferred.

[0033] The term "treating" or "treatment" of any disease or condition refers, in some embodiments, to ameliorating the disease or disorder (i.e., arresting or reducing the development of the disease or at least one of the clinical symptoms thereof). In some embodiments "treating" or "treatment" refers to ameliorating at least one physical parameter, which may not be discernible by the subject. In some embodiments, "treating" or "treatment" refers to modulating the disease or disorder, either physically, (e.g., stabilization of a discernible symptom), physiologically, (e.g., stabilization of a physical parameter), or both. In some embodiments, "treating" or "treatment" refers to delaying the onset of the disease or disorder, or even preventing the same. “Prophylactic treatment” is to be construed as any mode of treatment that is used to prevent progression of the disease or is used for precautionary purpose for persons at risk of developing the condition.

[0034] The term “pharmaceutically acceptable salts” means salts of compounds of the present invention which are pharmaceutically acceptable, as defined above, and which possess the desired pharmacological activity. Non-limiting examples of such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid; or with organic acids such as 1,2-ethanedisulfonic acid, 2-hydroxy ethanesulfonic acid, 2-naphthalenesulfonic acid, 3 -phenylpropionic acid, 4,4'-methylenebis(3-hydroxy- 2-ene- 1 -carboxylic acid), 4-methylbicyclo[2.2.2]oct-2-ene-l-carboxylic acid, acetic acid, aliphatic mono- and dicarboxylic acids, aliphatic sulfuric acids, aromatic sulfuric acids, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carbonic acid, cinnamic acid, citric acid, cyclopentanepropionic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, heptanoic acid, hexanoic acid, hydroxynaphthoic acid, lactic acid, laurylsulfuric acid, maleic acid, malicacid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid, o-(4-hydroxybenzoyl)benzoic acid, oxalic acid, / ?-chlorobenzenesulfonic acid, phenyl -substituted alkanoic acids, propionic acid, p-toluenesulfonic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, tartaric acid, tertiarybutyl acetic acid, and trimethylacetic acid. Pharmaceutically acceptable salts also include base addition salts which may be formed when acidic protons present are capable of reacting with inorganic or organic bases. Acceptable inorganic bases include sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide and calcium hydroxide. Non-limiting examples of acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, and / ' / -methylglucamine. It should be recognized that the particular anion or cation forming a part of any salt of this invention is not critical, so long as the salt, as a whole, is pharmacologically acceptable. Additional examples of pharmaceutically acceptable salts and their methods of preparation and use are presented in Handbook of Pharmaceutical Salts: Properties, and Use (P. H. Stahl & C. G. Wermuth eds., Verlag Helvetica Chimica Acta, 2002).

[0035] The ddhC is a naturally occurring nucleoside analogue produced by the viperin pathway. Studies have demonstrated the use of these compounds in reducing inflammatory cytokine IL-ip production in murine bone marrow derived macrophages (BMDMs). A hitherto undiscovered interaction has been identified between the nucleotide ddhCTP and pro-inflammatory enzyme CMPK2. Viperin KO cells showed higher titre of IL-ip compared with wild-type, indicating a relationship between viperin and reduction of IL-ip expression. Further, it is discovered that ddhCTP is capable of inhibiting the pro-inflammatory thymidylate kinase activity of CMPK2. In view of the activities of ddhCTP prodrugs, including ddhC nucleoside, in preventing secretion of IL-ip from “inflamed” macrophages, it is postulated that ddhC prodrugs inhibit NLRP3 inflammasome activation via the inhibition of CMPK2.

[0036] The compounds or ddhC prodrugs disclosed herein specifically target CMPK2- dependent NLRP3 inflammasome activation, which mediate chronic inflammatory illnesses, like gout and IBS. This design makes the compounds or ddhC prodrugs a much more precisely targeted anti- inflammatory, providing a good alternative treatment for patients with contraindications for the other anti-inflammatories like NSAIDs and corticosteroids. In addition, NLRP3 inflammasome has been a recent target in the treatment of breast cancer and severe CO VID-19.

[0037] An aspect of the patent document provides a compound of Formula I or a pharmaceutically acceptable salt thereof.Formula I wherein:M in each instance is independently O or S, m is 0, 1, or 2, n is 1, 2 or 3,X is selected from the group consisting of O, CH2, NH, CHF, CF2, CBr2, CCI2, and S,Y is selected from the group consisting of H, C1-12alkyl-C(O), C1-12alkyl-C(O)-O-C1-8alkylene-C(O), phenyl, phenyl-C(O), 5-6-membered heteroaryl-C(O), C1-6alkyl-O-C(O), C1-6alkyl-C(O)-O-C1-6alkylene, C1-6alkyl-O-C(O)-O-C1-6alkylene, P(O)(Ra)2, and P(S)(Ra)2, Rb-CH(N(RC)2)C(O),Wherein each Rais independently selected from the group consisting of OH, C1-12alkyl-C(O)-O-C1- salkylene-O, C1-6alkyl-O-C(O)-O-C1-4alkylene-O, Ci-2oalkyl-0-C1-8alkylene-0, Ci-2oalkyl-S-C1- 8alkylene-O, C1-12alkyl-C(O)C1-8alkylene-O, phenyl-O, naphthyl-O, 5-12-membered heteroaryl-O, Cn6alkyl-phenyl-O, C1-6alkyl-O-C(O)-C1-4alkylene-NH, C1-6alkyl-O-C(O)-CH(Rm)-NH, phenyl-O-C(O)- CH(Rm)-NH, heteroaryl-O-C(O)-CH(Rm)-NH, C1-8alkyl-C(O)-S-C1-8alkylene-O, C1-6alkyl-O-C(O)-S- C1-4alkylene-O, Ci-2oalkyl-S-C1-8alkylene-0, C1-6alkyl-S-C(O)-C1-4alkylene-NH, R°-C(O)-O-Ph-C1- 4alkylene-O, Ci-i8alkyl-O-CH2CH(ORp)CH2, Ci-i8alkyl-S-CH2CH(ORp)CH2, wherein the phenyl or naphthyl in Rais optionally substituted with one or more of halogen, C1-6alkyl, C3-6cycloalkyl, haloC1- ealkyl, CN, and OC1-6alkyl, wherein each Rmis independently a side chain (e.g., methyl, isopropyl, phenyl, etc.) of an amino acid, wherein each Rnis independently H or C1-6alkyl, wherein each R° is independently Ci-i8alkyl, C3- ecycloalkyl, C1-3alkylene-C3-6cycloalkyl, phenyl, 5-12-membered heteroaryl or PEG (50-500 Da), wherein each Rpis independently H, Ci-i8alkyl, Ci-i8alkyl-C(O), benzyl, or C2-18alkenyl, wherein Rbis a side chain of an amino acid, wherein each Rcis independently H or C1-6alkyl,Wherein the alkyl or alkylene of Y (e.g. C1-8alkyl of C1-8alkyl-C(O)) is optionally substituted with one or more of halogen, CN, OH, SH, (e.g. side chain of an amino acid), 5-10 membered heteroaryl, and phenyl, N(R°)2, wherein each R° is independently H or C1-8alkyl,wherein the phenyl and heteroaryl in Y are each optionally substituted with C1-4alkyl, CN, OH, halogen, and C1-4alkyl-O, one of R1and R2is H and the other is selected from the group consisting of H, OH, halogen, CN, OC1- ealkyl, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl,R3is selected from the group consisting of H, halogen, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl,R4and R5are each independently H, C1-4alkyl, or OH,R6is selected from the group consisting of H, CN, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl,A is selected from the group consisting of O, S, CH2, CF2, CCI2, CH(OH), and CHF;B is selected from the group consisting ofWherein Rd, and Reare each independently selected from the group consisting of H, C1-6alkyl, OH, OC1-6alkyl, C3-6cycloalkyl, Ci-salkylene-C3-6cycloalkyl, OC3-6cycloalkyl, phenyl, Ci-3alkylene-phenyl, NH, and NC1-6alkyl,Rfand Rgare each independently selected from H, C1-6alkyl, halo- C1-6alkyl, Ci-salkylene-OH, C1- 3alkylene-OC1-6alkyl, C3-6cycloalkyl, Ci-3alkylene-C3-6cycloalkyl, CN, halogen,R1is selected from H, C1-6alkyl, C3-6cycloalkyl, andRhis selected from the group consisting of H, C1-6alkyl, C3-6cycloalkyl, OH, OC1-6alkyl, NH, and NC1- ealkyl,E is selected from the group consisting of O, S, CH2, CHF, CF2, NH, CBr2, CR>Rk, and CCI2,R> andRkare independently selected from the group consisting of H, C1-6alkyl, halogen; alternatively Rj and Rklink up to form a 2-6 membered cycloalkyl ring.

[0038] The salt form of the compound may contain two cationic counter ions of charge 1+ or one cation of charge 2+ The cations can be metal cations or alkyl ammonium cations or other suitable forms. In some embodiments, the compound is in a dianionic salt form.

[0039] In some embodiments, R1and R2are each independently selected from H, OH, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl. In some embodiments, at least one of R1and R2is H. In some embodiments, one of R1and R2is H and the other is OH.

[0040] In some embodiments, R3is methyl, ethyl, propyl, C2-8alkenyl, C2-8alkynyl, F, or Cl. In some embodiments, R3is H.

[0041] In some embodiments, one of R4and R5is H and the other is OH or C1-6alkyl. In some embodiments, R4and R5are both H. In some embodiments, X is selected from CH2, CHF, CF2, CBr2, and CCI2.

[0042] In some embodiments, R6is selected from CN, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl. In some embodiments, R6is H.

[0043] In some embodiments, A is selected from O, S, CH2, CF2, CCI2, CH(OH), and CHF.

[0044] In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3.

[0045] In some embodiments, X is O. In some embodiments, X is S. In some embodiments, X is NH or NHC1-6alkyl.

[0046] In some embodiments, Y is H.

[0047] In some embodiments, Y is selected from C1-12alkyl-C(O), C1-12alkyl-C(O)-O-C1-8alkylene-C(O), phenyl-C(O), 5-6-membered heteroaryl -C(O), C1-6alkyl-O-C(O), C1-6alkyl-O-C(O)-O- C1-6alkylene, Rb-CH(N(RC)2) and Rb-CH(N(RC)2)C(O).

[0048] In some embodiments, Y is selected from the group consisting of C1-12alkyl-C(O), C1- i2alkyl-C(O)-O-Ci.8alkylene-C(O), C1-6alkyl-C(O)-O-C1-6alkylene, and C1-6alkyl-O-C(O)-O-C1- ealkylene.

[0049] In some embodiments, Y is phenyl-C(O), or 5-6-membered heteroaryl-C(O).

[0050] In some embodiments, Y is Rb-CH(N(RC)2). In some embodiments, Y is Rb- CH(N(RC)2)C(O).

[0051] In some embodiments, Y is P(O)(Ra)2 or P(S)(Ra)2. In some embodiments, at least one of the (Ra)2is OH. In some embodiments, one or both of the (Ra)2 are OH. In some embodiments, each Rais independently selected from the group consisting of OH, C1-12alkyl-C(O)-O-C1-8alkylene-O, C1- 6alkyl-O-C(O)-O-C1-4alkylene-O, C1-12alkyl-C(O)C1-8alkyle-O, Ci-2oalkyl-0-C1-8alkylene-0, C1-8alkyl- C(O)-S-C1-8alkylene-O, phenyl-O, 5-12-membered heteraryl-O, C1-6alkyl-phenyl-O, C1-6alkyl-O-C(O)-C1-4alkylene-NH, C1-6alkyl-O-C(O)-CH(Rm)-NH, phenyl -O-C(O)-CH(Rm)-NH, heteroaryl -O-C(O)- CH(Rm)-NH, Rm-CH(NRn)2).

[0052] In some embodiments, m is 1 or 2. In some embodiments, A is O.

[0053] In some embodiments, E in each instance is independently O or CH2.

[0054] In some embodiments, Y is P(O)(Ra)2 or P(S)(Ra)2.

[0055] In some embodiments, Y is P(O)(Ra)2, A is O, E is CH2, m is 1. d

[0056] In some embodiments,wherein R , Re, R and Rgare each independently selected from H, C1-6alkyl, OH, OC1-6alkyl, C3-6cycloalkyl, Ci-salkylene-C3-6cycloalkyl, phenyl, and Ci-3alkylene-phenyl, NH, and NC1-6alkyl. In some embodiments, Rd, Re, Rfand Rgare each H.RS ■w x Y

[0057] In some embodiments, B is °R, wherein R , Rgand R are each independently selected from H, C1-6alkyl, OH, OC1-6alkyl, C3-6cycloalkyl, C1-3alkylene-C3-6cycloalkyl, phenyl, and C1- salkylene-phenyl, NH, and NC1-6alkyl. In some embodiments, Rf, Rgand Rhare each H.

[0058] In some embodiments, Rdand Reare each independently selected from the group consisting of H, C1-6alkyl, OH, C3-6cycloalkyl, Ci-3alkylene-C3-6cycloalkyl, phenyl, and Ci-3alkylene- phenyl. In some embodiments, X is O or S. In some embodiments, n is 1 or 2. In some embodiments, A is O.

[0059] Nonlimiting examples of the base-containing moiety of the compounds of Formula I include the following

[0060] In some embodiments, Y is P(O)(Ra)2. In some embodiments, each Rais independently selected from the group consisting of OH, phenyl-O, naphthyl-O, C1-6alkyl-O-C(O)-CH(Rm)-NH, C1- 12alkyl-C(O)-O-C1-8alkylene-O, C1-6alkyl-O-C(O)-O-C1-4alkylene-O, Ci-2oalkyl-0-C1-8alkylene-0, C1- 2oalkyl-S-C1-8alkylene-0, C1-12alkyl-C(O)C1-8alkylene-O, R°-C(O)-O-Ph-C1-4alkylene-O, wherein the phenyl or naphthyl in Rais optionally substituted with one or more of halogen, C1-6alkyl, C3-6cycloalkyl, haloC1-6alkyl, CN, and OC1-6alkyl.

[0061] Nonlimiting examples of the compounds of Formula I include the following:

[0062] Rmand Rbin each instance are independently a side chain of an amino acid. Nonlimiting examples of the side chain include those of alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. Where one or more stereocenters are present in the compound, each stereocenter can be independently in the configuration of R or S. In some embodiments, the carbon where the side chain is attached has an S configuration. In some embodiments, the carbon where the side chain is attached has an R configuration.

[0063] In some embodiments, X is O, m is 0, one of the (Ra)2 is C1-6alkyl-O-C(O)-CH(Rm)-NH, and the other is phenyl-0 or naphthyl-O, wherein the phenyl or naphthyl is optionally substituted.

[0064] In some embodiments, m is 1 or 2, each E is independently selected from the group consisting of O, S, and CR>Rk, wherein R1and Rkare independently selected from the group consisting of H, C1-6alkyl, halogen, alternatively R> and Rklink up to form a 2-4 membered cycloalkyl ring.

[0065] In some embodiments, each E is independently selected from the group consisting of O,

[0066] In some embodiments, each Rais independently OH or R°-C(O)-O-Ph-C1-4alkylene-O, wherein the phenyl in Rais optionally substituted with one or more of halogen, C1-6alkyl, C3-6cycloalkyl, haloC1-6alkyl, CN, and OC1-6alkyl. In some embodiments, each Rais OH. In some embodiments, each Rais R°-C(O)-O-Ph-C1-4alkylene-O.

[0067] Nonlimiting examples of the compounds of Formula I include the following:

[0068] Additional examples of the compounds are selected from the following:

[0069] Further examples of the compounds of Formula I include the following

[0070] In some embodiments, the compound of Formula I is not one of the following

[0071] The compounds disclosed herein can be prepared by any suitable synthetic routes such as those illustrated in Figures 1-3. Additional procedures for compound synthesis are available in WO2022260535 and WO2022038539, the entire disclosure of which is hereby incorporated by reference.

[0072] Another aspect of the present disclosure provides a pharmaceutical composition containing a therapeutically effective amount of the above-described compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0073] The pharmaceutical composition may also contain one or more physiologically acceptable surface-active agents, additional carriers, diluents, excipients, smoothing agents, suspension agents, film forming substances, and coating assistants, or a combination thereof; and a compositiondisclosed herein. Acceptable additional carriers or diluents for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remington’s Pharmaceutical Sciences, 18th Ed., Mack Publishing Co., Easton, PA (1990), which is incorporated herein by reference in its entirety. Preservatives, stabilizers, dyes, sweeteners, fragrances, flavoring agents, and the like may be provided in the pharmaceutical composition. For example, sodium benzoate, ascorbic acid, and esters of p- hydroxybenzoic acid may be added as preservatives. In addition, antioxidants and suspending agents may be used. In various embodiments, alcohols, esters, sulfated aliphatic alcohols, and the like may be used as surface active agents; sucrose, glucose, lactose, starch, microcrystalline cellulose, crystallized cellulose, mannitol, light anhydrous silicate, magnesium aluminate, magnesium metasilicate aluminate, synthetic aluminum silicate, calcium carbonate, sodium acid carbonate, calcium hydrogen phosphate, calcium carboxymethyl cellulose, and the like may be used as excipients; magnesium stearate, talc, hardened oil and the like may be used as smoothing agents; coconut oil, olive oil, sesame oil, peanut oil, soya may be used as suspension agents or lubricants; cellulose acetate phthalate as a derivative of a carbohydrate such as cellulose or sugar, or methylacetate-methacrylate copolymer as a derivative of polyvinyl may be used as suspension agents; and plasticizers such as ester phthalates and the like may be used as suspension agents.

[0074] The pharmaceutical compounds described herein can be administered to a human patient per se, or in pharmaceutical compositions where they are mixed with other active ingredient(s), as in combination therapy, or suitable carriers or excipient(s). In some embodiments, a dosage form includes those forms in which the compound is administered per se. In addition, a dosage form may include a pharmaceutical composition. In any case, the dosage form may comprise a sufficient amount of the compound to treat a disease as part of a particular administration protocol, as would be understood by those of skill in the art. Techniques for formulation and administration of the compounds of the instant application may be found in “Remington’s Pharmaceutical Sciences,” Mack Publishing Co., Easton, PA, 18th edition, 1990.

[0075] The pharmaceutical compositions may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or tableting processes.

[0076] Pharmaceutical compositions may be formulated in any conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. Any of the well-known techniques,diluents, carriers, and excipients may be used as suitable and as understood in the art; e.g., in Remington’s Pharmaceutical Sciences, above.

[0077] Another aspect of this disclosure provides a method of treating a disease in a subject. The method includes administering to the subject in need a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt, isomer, or pharmaceutical composition thereo. Specific embodiments of the compound of Formula I are as described above.

[0078] The compounds or ddhC prodrugs disclosed herein can be applied to the treatment of a broad range of diseases or indications, which for the first time specifically target CMPK2-dependent NLRP3 inflammasome activation. Nonlimiting examples of the diseases or indications include gout, atherosclerosis, reperfusion injury, osteoarthritis, rheumatoid arthritis, COVID-19 and inflammatory bowel syndrome as well as neuropathologies (Alzheimer's disease), cancer, and chronic pain. The compounds or ddhC prodrugs may offer effective alternative treatments for settings where other anti- inflammatories are contraindicated. Although non-steroidal anti-inflammatory drugs (NSAIDs) are widely used, many patients with gastric ulcers or hypersensitivity / allergic reactions or allergic rhinitis to NSAIDs cannot take these drugs. The compounds disclosed herein provide an effective alternative to NSAIDs and fill a need that is not adequately addressed by corticosteroids (numerous side-effects and Cushing's Syndrome) and current biologies (exorbitant cost). In addition, the NLRP3 inflammasome has been a recent target in the treatment of breast cancer and is also implicated in gastric and head and neck cancer - there is evidence suggesting that NLRP3 inflammasome activation is necessary for the epithelial-mesenchymal transition and may contribute to alterations in the tumor microenvironment that allow for disease progression. NLRP3 is of widespread importance in the responses to viral pathogens and targeting or restricting NLRP3 inflammasome activity by treatment with ddhC might prevent or slow disease progression. In particular, severe COVID-19 is linked to uncontrolled inflammasome activation, including the NLRP3 inflammasome. Viral infection leads to rampant inflammation and the acute respiratory syndrome associated with COVID results from this uncontrolled inflammation. Enhanced intracellular ddhCTP levels that are driven by treatment with ddhC prodrug could limit viral proliferation as suggested in previous publications (i.e., target pathogen encoded proteins) and restrict the activation of the host's NLRP3 inflammasome, limiting the acute respiratory distress associated with COVID-19 infection.

[0079] Another aspect of the patent document provides a method of inhibiting CMPK2. The method includes contacting CMPK2 with an effective amount of the compound of formula I or its pharmaceutically acceptable salt. As a consequence of this inhibition, NLRP3 is inhibited as reflected in, for example, reduced maturation of IL-ip and IL- 18.

[0080] A related aspect of the patent document provides a method of inhibiting NLRP3. The method includes contacting CMPK2 with an effective amount of the compound of formula I or its pharmaceutically acceptable salt. In some embodiments, the contacting takes place in vivo. In some embodiments, the contacting takes place in a mamal or human.

[0081] Another aspect of the patent document provides a method of reducing or inhibiting maturation of IL-ip or IL-18, comprising contacting Cytidine / uridine monophosphate kinase 2 (CMPK2) with a therapeutically effective amount of a compound of Formula I. In some embodiments, the contacting takes place in vivo.

[0082] The compounds or pharmaceutical compositions described herein may be administered to the subject by any suitable means. Non-limiting examples of methods of administration include, among others, (a) administration though oral pathways, which administration includes administration in capsule, tablet, granule, spray, syrup, or other such forms; (b) administration through non-oral pathways such as rectal, vaginal, intraurethral, intraocular, intranasal, or intraauricular, which administration includes administration as an aqueous suspension, an oily preparation or the like or as a drip, spray, suppository, salve, ointment or the like; (c) administration via injection, subcutaneously, intraperitoneally, intravenously, intramuscularly, intradermally, intraorbitally, intracapsularly, intraspinally, intrastemally, or the like, including infusion pump delivery; as well as (d) administration topically; as deemed appropriate by those of skill in the art forbringing the active compound into contact with living tissue.

[0083] Pharmaceutical compositions suitable for administration include compositions where the active ingredients are contained in an amount effective to achieve its intended purpose. In some embodiments, a therapeutically effective amount of a compound is an amount effective to treat a viral infection, for example, in a mammalian subject (e.g., a human). The therapeutically effective amount of the compounds disclosed herein required as a dose will depend on the route of administration, the type of animal, including human, being treated, and the physical characteristics of the specific animal under consideration. The dose can be tailored to achieve a desired effect, but will depend on such factors as weight, diet, concurrent medication, and other factors which those skilled in the medical arts will recognize. More specifically, a therapeutically effective amount means an amount of compound effective to prevent, alleviate or ameliorate symptoms of disease or prolong the survival of the subject being treated. Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.

[0084] As will be readily apparent to one skilled in the art, the useful in vivo dosage to be administered and the particular mode of administration will vary depending upon the age, weight andmammalian species treated, the particular compounds employed, and the specific use for which these compounds are employed. The determination of effective dosage levels, that is the dosage levels necessary to achieve the desired result, can be accomplished by one skilled in the art using routine pharmacological methods. Typically, human clinical applications of products are commenced at lower dosage levels, with dosage level being increased until the desired effect is achieved. Alternatively, acceptable in vitro studies can be used to establish useful doses and routes of administration of the compositions identified by the present methods using established pharmacological methods.

[0085] In non-human animal studies, applications of potential products are commenced at higher dosage levels, with dosage being decreased until the desired effect is no longer achieved adverse side effects disappear. The dosage may range broadly, depending upon the desired effects and the therapeutic indication. Typically, dosages may be about 10 microgram / kg to about 100 mg / kg body weight, preferably about 100 microgram / kg to about 10 mg / kg body weight. Alternatively, dosages may be based and calculated upon the surface area of the patient, as understood by those of skill in the art.

[0086] The exact formulation, route of administration and dosage for the pharmaceutical compositions can be chosen by the individual physician in view of the patient’s condition, (see e.g., Fingl et al. 1975, in “The Pharmacological Basis of Therapeutics”, which is hereby incorporated herein by reference in its entirety, with particular reference to Ch. 1, p. 1). In some embodiments, the dose range of the composition administered to the patient can be from about 0.5 to about 1000 mg / kg of the patient’s body weight. The dosage may be a single one or a series of two or more given in the course of one or more days, as is needed by the patient. In instances where human dosages for compounds have been established for at least some conditions, those same dosages, or dosages that are about 0.1% to about 500%, more preferably about 25% to about 250% of the established human dosage may be used. Where no human dosage is established, as will be the case for newly discovered pharmaceutical compositions, a suitable human dosage can be inferred from ED50 or ID50 values, or other appropriate values derived from in vitro or in vivo studies, as qualified by toxicity studies and efficacy studies in animals.

[0087] It should be noted that the attending physician would know how to and when to terminate, interrupt, or adjust administration due to toxicity or organ dysfunctions. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response were not adequate (precluding toxicity). The magnitude of an administrated dose in the management of the disorder of interest will vary with the severity of the condition to be treated and to the route of administration. The severity of the condition may, for example, be evaluated, in part, by standard prognostic evaluation methods. Further, the dose and perhaps dose frequency will also vary according to the age, body weight,and response of the individual patient. A program comparable to that discussed above may be used in veterinary medicine.

[0088] Although the exact dosage will be determined on a drug-by-drug basis, in most cases, some generalizations regarding the dosage can be made. The daily dosage regimen for an adult human patient may be, for example, an oral dose of about 0.1 mg to 2000 mg of the active ingredient, preferably about 1 mg to about 500 mg, e.g. 5 to 200 mg. In other embodiments, an intravenous, subcutaneous, or intramuscular dose of the active ingredient of about 0.01 mg to about 100 mg, preferably about 0.1 mg to about 60 mg, e.g. about 1 to about 40 mg is used. In cases of administration of a pharmaceutically acceptable salt, dosages may be calculated as the free acid. In some embodiments, the composition is administered 1 to 4 times per day. Alternatively, the compositions may be administered by continuous intravenous infusion, preferably at a dose of up to about 1000 mg per day. As will be understood by those of skill in the art, in certain situations it may be necessary to administer the compounds disclosed herein in amounts that exceed, or even far exceed, the above-stated, preferred dosage range to effectively and aggressively treat particularly aggressive diseases or infections. In some embodiments, the compounds will be administered for a period of continuous therapy, for example for a week or more, or for months or years.

[0089] Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety, which are sufficient to maintain the antibiotic effects, or minimal effective concentration (MEC). The MEC will vary for each compound but can be estimated from in vitro data. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. However, HPLC assays or bioassays can be used to determine plasma concentrations.

[0090] Dosage intervals can also be determined using MEC value. Compositions should be administered using a regimen, which maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90% and most preferably between 50-90%.

[0091] In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.

[0092] The amount of composition administered may be dependent on the subject being treated, on the subject’s weight, the severity of the infection, the manner of administration and the judgment of the prescribing physician.

[0093] Compositions disclosed herein can be evaluated for efficacy and toxicity using known methods. For example, the toxicology of the compound may be established by determining in vitro toxicity towards a cell line, such as a mammalian, and preferably human, cell line. The results of suchstudies are often predictive of toxicity in animals, such as mammals, or more specifically, humans. Alternatively, the toxicity of particular compounds in an animal model, such as mice, rats, rabbits, or monkeys, may be determined using known methods. The efficacy of a particular compound may be established using several recognized methods, such as in vitro methods, animal models, or human clinical trials. Recognized in vitro models exist for nearly every class of condition. Similarly, acceptable animal models may be used to establish efficacy of chemicals to treat such conditions. When selecting a model to determine efficacy, the skilled artisan can be guided by the state of the art to choose an appropriate model, dose, and route of administration, and regime. Of course, human clinical trials can also be used to determine the efficacy of a compound in humans.

[0094] The compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, may be the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. Compositions comprising a compound formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0095] In some embodiments, in the pharmaceutical industry, it is standard practice to provide substantially pure material when formulating pharmaceutical compositions. Therefore, in some embodiments, “substantially pure” refers to the amount of purity required for formulating pharmaceuticals, which may include, for example, a small amount of other material that will not affect the suitability for pharmaceutical use. In some embodiments, the substantially pure compound contains at least about 96% of the compound by weight, such as at least about 97%, 98%, 99%, or 100% of the compound.

[0096] Examples

[0097] Example 1

[0098] This experiment examined the correlation between the presence of viperin and expression of IL-ip in cells. Viperin KO cells showed higher titre of IL-ip compared with wild-type, indicating a relationship between viperin and reduction of IL-ip expression. Cells were exposed to LPS.

[0099] The effect of ddhC and ddhC-protide and ddhC-isopropyl (marginal) on treated BMDMs (both wild-type and Viperin KO) were then investigated. ddhC and ddhC-protide were shown to drastically reduce IL-ip expression in both wild-type and viperin KO at Ihr and 12hr post intervention, with ddhC-isopropyl showing a reduction at the 12hr time point compared with the untreated control.

[0100] In addition to testing with ddhC, ddhC-protide and ddhC-isopropyl, ddhA was tested against the BMDM cell line at 1.2 mM. This resulted in cell death.

[0101] Example 2

[0102] Because ddhCTP is a unique inhibitor of CMPK2's kinase function, prodrugs of ddhCTP would be effective inhibitors of the NLRP3 dependent inflammasome. To test this hypothesis, immortalized murine bone marrow derived macrophages (iBMDMs) was utilized as a model for inflammation. When iBMDMs are treated with LPS (or other inflammatory signals), NLRP3 is expressed, whereupon a secondary signal, such as ATP, directs NLRP3 oligomerization, and the recruitment and activation of caspases. Active caspase are responsible for cleavage of pro-lL-ip to IL- ip, which is subsequently secreted to the extracellular milieu. Therefore, the ability of ddhC, which serves as a prodrug, was tested to restrict the release of IL-ip from iBMDM. Both wild-type iBMDMs and viperin knock-out iBMDMs were treated (which contain no ddhCTP) with DMSO or ddhC at 1 mM for 12 hr to allow for the intracellular formation and accumulation of ddhCTP. Subsequently, the media was removed, and fresh media was added that contained either DMSO or 1 mM ddhC and 400 ng of LPS. After 8 hr, 1 mM ATP was added, and the cells were further incubated for 40 min. Aliquots of media were immediately removed and tested for the presence of IL-ip by ELISA. As previously reported, with DMSO treatment, VipKO iBMDMs produce higher levels of IL-ip than wild-type iBMDMs, which is consistent with our hypothesis that viperin / ddhCTP inhibits / regulates CMPK2. Importantly, both wild-type and VipKO iBMDMs exhibit significantly reduced IL-ip levels when treated with ddhC. The magnitude of IL-ip reduction correlate with the intercellular concentration of ddhCTP within the iBMDMs (data showing that intracellular ddhCTP levels had an inverse correlation with IL-ip maturation, supporting the hypothesis that higher intracellular ddhCTP concentration leading to decreased IL-ip maturation. This behavior supports a mechanism in which the ddhC is converted to ddhCTP, which can effectively inhibit the kinase function of CMPK2 in a cell and thus restrict / reduce NLRP3 activation. The ELISA of IL-ip secretion including a transfection with poly dA:dT (synthetic double stranded DNA) was repeated, leading to activation of the AIM2 inflammasome, as well as with ATP, which activates the NLRP3 inflammasome. The Karin Lab previously established that AIM2 inflammasome activation is CMPK2 independent, thus AIM2 inflammasome mediated IL-ip should not be inhibited by ddhC treatment based on the hypothesis. These ELISA results demonstrate once againthat NLRP3 inflammasome activity was robustly inhibited in both WT and VipKO cells by ddhC treatment, whereas ddhC treatment had no statistically significant effect on AIM2 inflammasome activity. These data support the hypothesis that CMPK2 inhibition will likely be a broad-spectrum and selective anti-inflammatory strategy against NLRP3 -dependent diseases and ddhC prodrug strategies will be effective and safe.

[0103] All references cited herein are incorporated herein by reference in their entireties. It will be appreciated by persons skilled in the art that the present invention is not limited to what has been particularly shown and described. Rather, the scope of the present invention is defined by the claims which follow. It should further be understood that the above description is only representative of illustrative examples of embodiments. The description has not attempted to exhaustively enumerate all possible variations. The alternate embodiments may not have been presented for a specific portion of the invention, and may result from a different combination of described portions, or that other un- described alternate embodiments may be available for a portion, is not to be considered a disclaimer of those alternate embodiments. It will be appreciated that many of those un-described embodiments are within the literal scope of the following claims, and others are equivalent.

Claims

WE CLAIM1. A method of treating a disease associated with nod-like receptor protein 3 (NLRP3) inflammasome activation, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof,Formula IWherein:M in each instance is independently O or S, m is 0, 1, or 2, n is 1, 2 or 3,X is selected from the group consisting of O, CH2, NH, CHF, CF2, CBr2, CCI2, and S,Y is selected from the group consisting of H, C1-12alkyl-C(O), C1-12alkyl-C(O)-O-C1- 8alkylene-C(O), phenyl, phenyl-C(O), 5-6-membered heteroaryl-C(O), C1-6alkyl-O-C(O), C1-6alkyl-C(O)-O-C1-6alkylene, C1-6alkyl-O-C(O)-O-C1-6alkylene, P(O)(Ra)2, and P(S)(Ra)2, Rb-CH(N(RC)2)C(O),Wherein each Rais independently selected from the group consisting of OH, C1- i2alkyl-C(O)-O-C1-8alkylene-O, C1-6alkyl-O-C(O)-O-C1-4alkylene-O, Ci-2oalkyl-0-C1- 8alkylene-O, Ci-2oalkyl-S-C1-8alkylene-0, C1-12alkyl-C(O)C1-8alkylene-O, phenyl-O, naphthyl-O, 5-12-membered heteroaryl-O, C1-6alkyl-phenyl-O, C1-6alkyl-O-C(O)-Cn 4alkylene-NH, C1-6alkyl-O-C(O)-CH(Rm)-NH, phenyl-O-C(O)-CH(Rm)-NH, heteroaryl-O-C(O)-CH(Rm)-NH, C1-8alkyl-C(O)-S-C1-8alkylene-O, C1-6alkyl-O-C(O)- S-C1-4alkylene-O, Ci-2oalkyl-S-C1-8alkylene-0, C1-6alkyl-S-C(O)-C1-4alkylene-NH, R°- C(0)-0-Ph-C1-4alkylene-0 (bisphosphonate), C1-18alkyl-O-CH2CH(ORp)CH2, Cn33 ctive\106465225.vl-l / 16 / 20173631489.2i8alkyl-S-CH2CH(ORp)CH2, wherein the phenyl or naphthyl in Rais optionally substituted with one or more of halogen, C1-6alkyl, C3-6cycloalkyl, haloC1-6alkyl, CN, and OC1-6alkyl, wherein each Rmis independently a side chain of an amino acid, wherein each Rnis independently H or C1-6alkyl, wherein each R° is independently Ci-isalkyl, C3- ecycloalkyl, Ci-salkylene-C3-6cycloalkyl, phenyl, 5-12-membered heteroaryl or PEG (50-500 Da), wherein each Rpis independently H, Ci-isalkyl, Ci-i8alkyl-C(O), benzyl, or C2-i8alkenyl, wherein Rbis a side chain of an amino acid, wherein each Rcis independently H or Ci- ealkyl,Wherein the alkyl or alkylene of Y (e.g. C1-8alkyl of C1-8alkyl-C(O)) is optionally substituted with one or more of halogen, CN, OH, SH, (e.g. side chain of an amino acid), 5-10 membered heteroaryl, and phenyl, N(R°)2, wherein each R° is independently H or C1-8alkyl, wherein the phenyl and heteroaryl in Y are each optionally substituted with C1-4alkyl, CN, OH, halogen, and C1-4alkyl-O, one of R1and R2is H and the other is selected from the group consisting of H, OH, halogen, CN, OC1-6alkyl, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl,R3is selected from the group consisting of H, halogen, C1-6alkyl, C2-6alkenyl, and C2- ealkynyl,R4and R5are each independently H, C1-4alkyl,R6is selected from the group consisting of H, CN, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl,A is selected from the group consisting of O, S, CH2, CF2, CCI2, CH(OH), and CHF;B is selected from the group consisting ofWherein Rd, and Reare each independently selected from the group consisting of H, C1- ealkyl, OH, OC1-6alkyl, C3-6cycloalkyl, Ci-salkylene-C3-6cycloalkyl, OC3-6cycloalkyl, phenyl, Ci-3alkylene-phenyl, NH, and NC1-6alkyl,Rfand Rgare each independently selected from H, C1-6alkyl, halo- C1-6alkyl, Ci-3alkylene- OH, Ci-3alkylene-OC1-6alkyl, C3-6cycloalkyl, Ci-3alkylene-C3-6cycloalkyl, CN, halogen,R1is selected from H, C1-6alkyl, C3-6cycloalkyl, andRhis selected from the group consisting of H, C1-6alkyl, C3-6cycloalkyl, OH, OC1-6alkyl, NH, and NC1-6alkyl,E is selected from the group consisting of O, S, NH, and CR'Rk,R> andRkare independently selected from the group consisting of H, C1-6alkyl, halogen; alternatively Rjand Rklink up to form a 2-6 membered cycloalkyl ring.

2. The method of claim 1, wherein Y is selected from the group consisting of C1-12alkyl-C(O), C1-12alkyl-C(O)-O-C1-8alkylene-C(O), phenyl-C(O), 5-6-membered heteroaryl-C(O), C1- 6alkyl-O-C(O), Ci.6alkyl-O-C(O)-O-Ci.6alkylene, P(O)(Ra)2and P(S)(Ra)2.

3. The method of any one of claims 1-2, wherein wherein4. The method of any one of claims 1-3, wherein Rd, and Reare each independently selected from the group consisting of H, C1-6alkyl, OH, C3-6cycloalkyl, Ci-salkylene-C3-6cycloalkyl, phenyl, and Ci-3alkylene-phenyl.

5. The method of any one of claims 1-4, wherein X is O or S.

6. The method of any one of claims 1-4, wherein A is O.

7. The method of any one of claims 1-5, wherein n is 1 or 2.

8. The method of any one of claims 1-5, wherein Y is P(0)(Ra)29. The method of any one of claims 1-5, wherein each Rais independently selected from the group consisting of OH, phenyl-O, naphthyl-O, C1-6alkyl-O-C(O)-CH(Rm)-NH, Ci-nalkyl- C(O)-O-C1-8alkylene-O, C1-6alkyl-O-C(O)-O-C1-4alkylene-O, Ci-2oalkyl-0-C1-8alkylene-0, Ci-2oalkyl-S-C1-8alkylene-0, C1-12alkyl-C(O)C1-8alkylene-O, R°-C(O)-O-Ph-C1-4alkylene-O, wherein the phenyl or naphthyl in Rais optionally substituted with one or more of halogen, C1-6alkyl, C3-6cycloalkyl, haloC1-6alkyl, CN, and OC1-6alkyl.

10. The method of any one of claims 1-5, wherein X is O, m is 0, one of the (Ra)2 is C1-6alkyl- O-C(O)-CH(Rm)-NH, and the other is phenyl-0 or naphthyl-O, wherein the phenyl or naphthyl is optionally substituted.

11. The method of any one of claims 1-9, wherein m is 1 or 2, each E is independently selected from the group consisting of O, S, and CR'Rk, wherein R1andRkare independently selected from the group consisting of H, C1-6alkyl, halogen, alternatively R1and Rklink up to form a 2-4 membered cycloalkyl ring.

12. The method of claim 11, wherein each E is independently selected from the group consisting13. The method of any one of claims 11-12, wherein each Rais independently OH or R°-C(O)- O-Ph-C1-4alkylene-O, wherein the phenyl in Rais optionally substituted with one or more of halogen, C1-6alkyl, C3-6cycloalkyl, haloC1-6alkyl, CN, and OC1-6alkyl.

14. The method of any one of claims 11-12, wherein m is 1, each Rais independently OH.

15. The method of claim 1, wherein the compound is selected from the group consisting of16. The method of claim 1, wherein the inflammatory disease is selected from the group consisting of gout, inflammatory bowel disease, rheumatoid arthritis, atherosclerosis, reperfusion injury, osteoarthritis, neurodegenerative disease (e.g. Alzheimer’s disease),Stargardt disease type 1, cryopyrin-associated periodic syndrome (CAPS), acute inflammatory demyelinating polyradiculoneuropathy, autoinflammatory syndromes, Behcet disease, breast cancer, gastric cancer, head and neck cancer, nasopharyngeal carcinoma, NSCLC, cervical cancer, glioma, ovarian cancer, lung canacer, acute myeloid leukemai, chronic myeloid leukemia, liver cancer, osteocarcoma, renal cell carcinoma, oral cancer, prostate cancer, and drug addiction.

17. A method of inhibiting NLRP3 inflammasome activation, comprising contacting Cytidine / uridine monophosphate kinase 2 (CMPK2) with a therapeutically effective amount of the compound of Formula I.

18. A method of inhibiting inhibiting maturation of IL-ip or IL-18, comprising contacting Cytidine / uridine monophosphate kinase 2 (CMPK2) with a therapeutically effective amount of a compound of Formula I.

19. A compound of Formula I or a pharmaceutically acceptable salt thereof,Formula IWherein:M in each instance is independently O or S, m is 0, 1, or 2, n is 1, 2 or 3,X is selected from the group consisting of O, CH2, NH, CHF, CF2, CBr2, CCI2, and S,Y is selected from the group consisting of H, C1-12alkyl-C(O), C1-12alkyl-C(O)-O-C1- salkylene-C(O), phenyl, phenyl-C(O), 5-6-membered heteroaryl-C(O), C1-6alkyl-O-C(O), C1-6alkyl-C(O)-O-C1-6alkylene, C1-6alkyl-O-C(O)-O-C1-6alkylene, P(O)(Ra)2, and P(S)(Ra)2, Rb-CH(N(RC)2)C(O),Wherein each Rais independently selected from the group consisting of OH, C1- i2alkyl-C(O)-O-C1-8alkylene-O, C1-6alkyl-O-C(O)-O-C1-4alkylene-O, Ci-2oalkyl-0-C1-salkylene-O, Ci-2oalkyl-S-C1-8alkylene-0, C1-12alkyl-C(O)C1-8alkylene-O, phenyl-O, naphthyl-O, 5-12-membered heteroaryl-O, C1-6alkyl-phenyl-O, C1-6alkyl-O-C(O)-C1- 4alkylene-NH, C1-6alkyl-O-C(O)-CH(Rm)-NH, phenyl -O-C(O)-CH(Rm)-NH, heteroaryl -O-C(O)-CH(Rm)-NH, C1-8alkyl-C(O)-S-C1-8alkylene-O, C1-6alkyl-O-C(O)- S-C1-4alkylene-O, Ci-2oalkyl-S-C1-8alkylene-0, C1-6alkyl-S-C(O)-C1-4alkylene-NH, R°- C(O)-O-Ph-C1-4alkylene-O, Ci-i8alkyl-O-CH2CH(ORp)CH2, Ci-isalkyl-S- CH2CH(ORP)CH2, wherein the phenyl or naphthyl in Rais optionally substituted with one or more of halogen, C1-6alkyl, C3-6cycloalkyl, haloC1-6alkyl, CN, and OC1-6alkyl, wherein each Rmis independently a side chain of an amino acid, wherein each Rnis independently H or C1-6alkyl, wherein each R° is independently Ci-i8alkyl, C3- ecycloalkyl, Ci-salkylene-C3-6cycloalkyl, phenyl, 5-12-membered heteroaryl or PEG (50-500 Da), wherein each Rpis independently H, Ci-i8alkyl, Ci-i8alkyl-C(O), benzyl, or C2-i8alkenyl, wherein Rbis a side chain of an amino acid, wherein each Rcis independently H or C1- ealkyl,Wherein the alkyl or alkylene of Y (e.g. C1-8alkyl of C1-8alkyl-C(O)) is optionally substituted with one or more of halogen, CN, OH, SH, (e.g. side chain of an amino acid), 5-10 membered heteroaryl, and phenyl, N(R°)2, wherein each R° is independently H or C1-8alkyl, wherein the phenyl and heteroaryl in Y are each optionally substituted with C1-4alkyl, CN, OH, halogen, and C1-4alkyl-O, one of R1and R2is H and the other is selected from the group consisting of H, OH, halogen, CN, OC1-6alkyl, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl,R3is selected from the group consisting of H, halogen, C1-6alkyl, C2-6alkenyl, and C2- ealkynyl,R4and R5are each independently H or C1-4alkyl,R6is selected from the group consisting of H, CN, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl,A is selected from the group consisting of O, S, CH2, CF2, CCI2, CH(OH), and CHF;B is selected from the group consisting ofWherein Rd, and Reare each independently selected from the group consisting of H, C1- ealkyl, OH, OC1-6alkyl, C3-6cycloalkyl, C1-3alkylene-C3-6cycloalkyl, OC3-6cycloalkyl, phenyl, Ci-3alkylene-phenyl, NH, and NC1-6alkyl,Rfand Rgare each independently selected from H, C1-6alkyl, halo- C1-6alkyl, C1-3alkylene- OH, Ci-3alkylene-O C1-6alkyl, C3-6cycloalkyl, C1-3alkylene-C3-6cycloalkyl, CN, halogen,R1is selected from H, C1-6alkyl, C3-6cycloalkyl, andRhis selected from the group consisting of H, C1-6alkyl, C3-6cycloalkyl, OH, OC1-6alkyl, NH, and NC1-6alkyl,E is selected from the group consisting of O, S, NH, and CRjRk,R> andRkare independently selected from the group consisting of H, C1-6alkyl, halogen; alternatively Rjand Rklink up to form a 2-6 membered cycloalkyl ring. provided that the compound is not one of the following:

20. A pharmaceutical composition comprising a therapeutically effective amount of the compound or the pharmaceutically acceptable salt thereof of claim 19.

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