Small molecule degraders of soluble epoxide hydrolase

Abstract

This patent document discloses small molecule degraders of an enzyme, soluble epoxide hydroxlase (sEH). Also provided are pharmaceutical compositions of the small molecules and methods of treating sEH associated diseases.

Description

182219.00268 SMALL MOLECULE DEGRADERS OF SOLUBLE EPOXIDE HYDROLASE STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0001] This invention was made with government support under ES030443, and ES004699 awarded by the National Institutes of Health. The government has certain rights in the invention. TECHNICAL FIELD

[0002] This patent document discloses small molecule degraders of an enzyme, soluble epoxide hyrolase (sEH). These molecules that bind to the enzyme are conjugated to a backbone that promotes degradation of the enzyme via the ubiquitination pathway. BACKGROUND

[0003] Soluble epoxide hydrolase (sEH) is an enzyme responsible for the conversion of epoxy fatty acids (EpFAs) into their corresponding vicinal diols in the arachidonic acid cascade. These lipid mediators are important endogenous signaling molecules in glucose homeostasis, vascular regulation and pain. Inhibition of sEH hydrolase activity by small molecule she inhibitors (sEHi) is considered a promising approach for the treatment of various diseases such as diabetes, neuropathic pain, chronic obstructive pulmonary disease (COPD), and metabolic disorders. Cellular mechanistic studies showed that the beneficial effects of sEHi are resulted from the ER stress reduction caused by the increase of the EpFAs. Currently, there is no sEH inhibitors in clinical use yet. The potential reason is that traditional, occupancy- driven inhibitors are unable to completely block the enzyme function invivo. This problem arises because (a) sEH protein level is high, reaching up to sub μM in the liver fraction, necessitating a high concentration of inhibitor required to show efficacy even with nM affinity inhibitors, (b) extremely high occupancy of catalytic site is required to block the conversion of endogenous substrate.

[0004] A need exists to develop new therapeutic agents for treating diseases and conditions associated with sEH. SUMMARY

[0005] This patent document discloses small-molecule sEH degraders based on PROTAC technology. The molecules exhibit high potency with excellent stability. 1 171560744182219.00268

[0006] An aspect of this patent document provides a compound or a pharmaceutically acceptable salt thereof, wherein the compound is represented by Formula I,Formula I wherein A is a C3-6 cycloalkyl, a phenyl, 5- or 6-membered heteroaryl, benzyl or, wherein A is optionally substituted with one or more substituents selected from the group consisting of O- haloC1-4alkyl, phenyl, 5- or 6-membered heteroaryl, haloC1-4alkyl, C1-4alkyl, 3-6 membered-cycloalkyl, OC1-4 alkyl, O-haloC1-4 alkyl, OH, CN, halogen, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; B is a carbonyl (C=O) or an amide, wherein the carbonyl carbon of the amide is bonded to L1; C is an amide, a carbonyl (C=O), or OC=O, wherein the carbonyl carbon of the amide or the OC=O is bonded to (L3)m; D is a bond or a bicyclic ring or a single aromatic ring, wherein the ring is optionally substituted with one or more substituents selected from the group consisting of haloOC1- 4alkyl, haloC1-4 alkyl, C1-4 alkyl, 3-6 membered-cycloalkyl, OC1-4 alkyl, OH, CN, halogen, NRcRd, wherein Rcand Rdin each instance is independently H or C1-4alkyl;optionally substituted with one or more substituents selected from the group consisting of haloOC1-4alkyl, haloC1-4alkyl, C1-4alkyl, OC1-4 alkyl, 3-6 membered-cycloalkyl, OH, CN, halogen, NRcRd, wherein Rcand Rdin each instance is independently H or C1-4alkyl; L1is a bond or a linker comprising 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, NH- 3-6 membered cycloalkyl, or NH-3-6 membered heterocycloalkyl; L2is a bond or a linker comprising one or more of phenyl, 5-6 membered heteroaryl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, C1-6alkylene, O, C(O), NH, and NC1-6alkyl, wherein the phenyl, 5-6 membered heteroaryl, 3-6 membered cycloalkyl, and 3-6 membered heterocycloalkyl are each optionally substituted with one or more of O-haloC1-4 alkyl, haloC1-4alkyl, C1-4alkyl, CN, and halogen; L3is a bond or a linker comprising one or more of phenyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, bicyclic ring, tricyclic ring, C1-6 alkylene, C2-6 alkenlene, C2-6 2 171560744182219.00268 alkynlene, O, S, C(O), NH, (OCH2CH2)1-20, NC1-6alkyl, and any combination thereof (each of the atoms or groups can be used multiple times in L3, and m is an integer from 1 to 20; Alternatively, D-E together is selected from the group consisting ofL1is a bond or a linker comprising 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, NH- 3-6 membered cycloalkyl, or NH-3-6 membered heterocycloalkyl; L2is a bond or a linker comprising one or more of phenyl, 5-6 membered heteroaryl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, C1-6 alkylene, O, C(O), NH, and NC1-6alkyl, wherein the phenyl, 5-6 membered heteroaryl, 3-6 membered cycloalkyl, and 3-6 membered heterocycloalkyl are each optionally substituted with one or more of O-haloC1-4alkyl, haloC1-4 alkyl, C1-4 alkyl, CN, and halogen; L3is a bond or a linker comprising one or more of phenyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, bicyclic ring, tricyclic ring, C1-6alkylene, C2-6alkenlene, C2-6alkynlene, O, S, C(O), NH, (OCH2CH2)1-20, NC1-6alkyl, and any combination thereof (each of the atoms or groups can be used multiple times in L3, and m is an integer from 1 to 20; 3 171560744182219.00268 provided that the compound is notalternatively,, wherein R1and R2in each instance are independently selected from the group consisting of haloOC1-4 alkyl, haloC1-4 alkyl, C1- 4alkyl, OC1-4 alkyl, OH, CN, halogen, NReRd, wherein Reand Rfin each instance is independently H or C1-4alkyl, and L1-L2is a linker comprising C1-8alkylene, 3-6 membered 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl; n and p are independently 0, 1, 2, or 3; Alternatively, D-E together is selected from the group consisting of ,Wherein R3is selected from the group consisting of H, C1-4alkyl, and C(O)C1-4alkyl, R4is selected from the group consisting of H, phenyl and benzyl, wherein the phenyl and benzyl is optionally substituted with one or more substituents selected from the group consisting of haloC1-4alkyl, C1-4alkyl, OC1-4alkyl, OH, CN, and halogen, Wherein R5is selected from the group consisting of haloC1-4 alkyl, C1-4 alkyl, OC1-4 alkyl, OH, CN, and halogen, wherein q is 0, 1, 2, 3 or 4; 4 171560744182219.00268 Wherein R6is selected from the group consisting of H, C1-4 alkyl, CN, and halogen, wherein r is is 0, 1, 2, 3 or 4; Wherein R7is selected from the group consisting of H, C1-4alkyl, CN, and halogen, R8is H, or C1-4 alkyl, wherein s is is 0, 1, 2, 3 or 4; Wherein R9is selected from the group consisting of H, halogen, C1-4 alkyl, CN, and halogen, R10is selected from the group consisting of H, halogen, C1-4alkyl, and CN,, wherein t is 0, 1, 2, 3 or 4; Wherein u is 0, 1, 2, 3 or 4.

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

[0009] Another aspect provides a method of degrading a sEH, comprising contacting sEH with a therapeutically effective amount of the compound disclosed herein or pharmaceutically acceptable salt thereof.

[0010] DESCRIPTIONS OF DRAWINGS

[0011] Figure 1 provides example compounds under Formula I.

[0012] Figure 2 provides example compounds under Formula I.

[0013] Figure 3 provides example compounds under Formula I.

[0014] Figure 4 provides example compounds under Formula I.

[0015] Figure 5 provides example compounds under Formula I.

[0016] Figure 6 provides example compounds under Formula I.

[0017] Figure 7 provides example compounds under Formula I.

[0018] Figure 8 provides example compounds under Formula I.

[0019] Figure 9 provides example compounds under Formula I.

[0020] Figure 10 provides example compounds under Formula I.

[0021] Figure 11 provides example compounds under Formula I.

[0022] Figure 12 provides example compounds under Formula I. 5 171560744182219.00268

[0023] Figure 13 provides example compounds under Formula I.

[0024] Figure 14 provides example compounds under Formula I.

[0025] Figure 15 provides example compounds under Formula I.

[0026] Figure 16 provides example compounds under Formula I.

[0027] Figure 17 provides example compounds under Formula I.

[0028] Figure 18 provides example compounds under Formula I.

[0029] Figure 19 provides example compounds under Formula I.

[0030] Figure 20 provides example compounds under Formula I.

[0031] Figure 21 provides example compounds under Formula I.

[0032] Figure 22 provides example compounds under Formula I.

[0033] Figure 23 provides example compounds under Formula I.

[0034] Figure 24 provides example compounds under Formula I.

[0035] Figure 25 provides example compounds under Formula I.

[0036] Figure 26 provides example compounds under Formula I. DETAILED DESCRIPTION

[0037] Various embodiments of this patent document potent and stable proteolysis targeting chimera (PROTAC) compounds targeting sEH. These compounds find use in the treatment of various diseases and conditions including diabetic neuropathic pain, chronic obstructive pulmonary disease (COPD), and Alzheimer disease.

[0038] 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.

[0039] 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. 6 171560744182219.00268

[0040] 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.

[0041] 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.

[0042] The term “carrier” refers to a chemical compound that facilitates the incorporation of a compound into cells or tissues.

[0043] The term "degrade" and "degradation" refers to the reduction of the amount of sEH in a cell.

[0044] 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.

[0045] 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.

[0046] 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. 7 171560744182219.00268

[0047] 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 "C1-10 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 " C1-4 alkyl" refers to alkyl groups having 1, 2, 3, or 4 carbon atoms. 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.

[0048] 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 CH2, (CH2)2, CH2CH(CH3), and the like. A C1-6 alkylene has 1, 2, 3, 4, 5 or 6 carbons. A C1-4alkylene has 1, 2, 3 or 4 carbons.

[0049] The term "alkenlene” refers to a double-bond containing divalent hydrocarbon which may be either straight-chained or branched. Non-limiting examples include CH=CH, CH2CH=CH, CH2CH=C(CH3), CH2CH2C(CH3)=C(CH3)CH2.

[0050] The term " alkynlene” refers to a triple-bond containing divalent hydrocarbon which may be either straight-chained or branched. Non-limiting examples include C≡C, CH2C≡C, CH2C≡CCH2, and CH2CH2CH(CH3)C≡CHCH2.

[0051] The terms of Cx-yalkylene, Cx-yalkenlene, and Cx-yalkynlene can also be described as alkylenex-y, alkenlenex-y, and alkynlenex-y, respectively. When x-y is 1 to 20, for example, the number of carbons can be any in the range, including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20.

[0052] The term “C1-4 alkoxy” includes an alkyloxy group having 1, 2, 3 or 4 carbons.

[0053] 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. 8 171560744182219.00268

[0054] The term “haloalkyl” refers to a C1-10 alkyl, straight chain or branched, in which one or more hydrogen has been replaced by a halogen. Non-limiting examples of haloalkyls include CHF2, CFH2, CF3, CF2CH3, CH2CF3, and CH2CH2F. In some embodiments, the alkyl in haloalkyl has 1, 2, 3 or 4 carbons. Likewise, the term “haloalkylene” refers to an alkylene, straight or branched, in which one or more hydrogen has been replaced by a halogen. Non- limiting examples of haloalkylenes include CHF, CF2, and CH2CF2.

[0055] The term “heterocycle” or "heterocycloalkyl" refers to 3 to 10 membered substituted or non-substituted non-aromatic cyclic groups where one or more carbon ring atoms are replaced with hetero atoms or groups containing heteroatoms (e.g. NH, NC1-4alkyl O, and S). Nonlimiting examples include pyrrolidine, piperidine, piperazine, N-methyl-piperazine, and morpholine. Optional substituents include C1-6 alkyl, C1-4 alkoxy, halogen, haloalkyl, sulfonamido, and amido.

[0056] The term “bicyclic” or “tricyclic” includes two or three rings joined together in a form of a fused ring system, bridged ring system, and a spiro ring system. One or more carbon ring atoms can be replaced with a heteroatom.

[0057] 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, acephenanthrene, anthracene, azulene, benzene, fluoranthene, fluorene, hexalen, hexaphene, hexylene, as-indacene, s-indacene, indane, indene, naphthalene, astacene, octaphene, octylene, 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.

[0058] 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.

[0059] The term "halogen" refers to F, Cl, Br, or I.

[0060] The term “carboxamide” refers to a group of -CONRR, wherein each R is independently a hydrogen, C1-6alkyl, 3-7 membered carbocycle, 3-7 membered heterocycle, 5- 10 membered heteroaryl or 6-10 membered aryl. The two R groups may link up to form a 3-7 9 171560744182219.00268 membered carbocycle, 3-7 membered heterocycle, 5-10 membered heteroaryl or 6-10 membered aryl.

[0061] 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 π electrons shared in a cyclic array, wherein at least one ring atom contributing to the shared π 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.

[0062] The term "subject" encompasses any animal, but preferably a mammal, e.g., a human, a non-human primate, a dog, a cat, a horse, a cow, or a rodent. More preferably, the subject is a human.

[0063] 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.

[0064] 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, 10 171560744182219.00268 2-hydroxyethanesulfonic 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- 1-carboxylic acid, acetic acid, aliphatic mono- and dicarboxylic acids, aliphatic sulfuric acids, aromatic sulfuric acids, benzenesulfonic acid, benzoic acid, camphor sulfonic 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, malic acid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid, o-(4-hydroxybenzoyl)benzoic acid, oxalic acid, p-chlorobenzenesulfonic acid, phenyl-substituted alkanoic acids, propionic acid, p-toluenesulfonic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, tartaric acid, tertiarybutylacetic 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 N-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).

[0065] Soluble epoxide hydrolase (sEH) is an enzyme responsible for the conversion of epoxy fatty acids (EpFAs) into their corresponding vicinal diols in the arachidonic acid cascade. These lipid mediators are important endogenous signaling molecules in glucose homeostasis, vascular regulation and pain. Inhibition of sEH hydrolase activity by small molecule sEH degraders based on PROTAC technology is a new approach for the treatment of various diseases such as diabetes, neuropathic pain, chronic obstructive pulmonary disease (COPD), and metabolic disorders.

[0066] An aspect of this patent document provides a compound or a pharmaceutically acceptable salt thereof, wherein the compound is represented by Formula I,Formula I 11 171560744182219.00268 wherein A is a C3-6cycloalkyl, a phenyl, 5- or 6-membered heteroaryl, benzyl or , wherein A is optionally substituted with one or more substituents selected from the group consisting of O- haloC1-4 alkyl, phenyl, 5- or 6-membered heteroaryl, haloC1-4 alkyl, C1-4 alkyl, 3-6 membered-cycloalkyl, OC1-4alkyl, O-haloC1-4alkyl, OH, CN, halogen, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; B is a carbonyl (C=O) or an amide, wherein the carbonyl carbon of the amide is bonded to L1; C is an amide, a carbonyl (C=O), or OC=O, wherein the carbonyl carbon of the amide or the OC=O is bonded to (L3)m; D is a bond or a bicyclic ring or a single aromatic ring, wherein the ring is optionally substituted with one or more substituents selected from the group consisting of haloOC1-4alkyl, haloC1-4alkyl, C1-4alkyl, 3-6 membered-cycloalkyl, OC1-4alkyl, OH, CN, halogen, NRcRd, wherein Rcand Rdin each instance is independently H or C1-4 alkyl;optionally substituted with one or more substituents selected from the group consisting of haloOC1-4alkyl, haloC1-4 alkyl, C1-4 alkyl, OC1-4alkyl, 3-6 membered-cycloalkyl, OH, CN, halogen, NRcRd, wherein Rcand Rdin each instance is independently H or C1-4alkyl; L1is a bond or a linker comprising 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, NH- 3-6 membered cycloalkyl, or NH-3-6 membered heterocycloalkyl; L2is a bond or a linker comprising one or more of phenyl, 5-6 membered heteroaryl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, C1-6 alkylene, O, C(O), NH, and NC1- 6alkyl, wherein the phenyl, 5-6 membered heteroaryl, 3-6 membered cycloalkyl, and 3-6 membered heterocycloalkyl are each optionally substituted with one or more of O-haloC1-4alkyl, haloC1-4 alkyl, C1-4 alkyl, CN, and halogen; L3is a bond or a linker comprising one or more of phenyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, bicyclic ring, tricyclic ring, C1-6alkylene, C2-6alkenlene, C2-6alkynlene, O, S, C(O), NH, (OCH2CH2)1-20, NC1-6alkyl, and any combination thereof (each of the atoms or groups can be used multiple times in L3, and m is an integer from 1 to 20; 12 171560744182219.00268 alternatively,, wherein R1and R2in each instance are independently selected from the group consisting of haloOC1-4 alkyl, haloC1-4 alkyl, C1-4 alkyl, OC1-4 alkyl, OH, CN, halogen, NReRd, wherein Reand Rfin each instance is independently H or C1-4alkyl, and L1-L2is a linker comprising C1-8alkylene, 3-6 membered 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl, and n and p are independently 0, 1, 2, or 3; Alternatively, D-E together is selected from the group consisting of13 171560744182219.0026814 171560744182219.00268, Wherein R3is selected from the group consisting of H, C1-4alkyl, and C(O)C1-4alkyl, R4is selected from the group consisting of H, phenyl and benzyl, wherein the phenyl and benzyl is optionally substituted with one or more substituents selected from the group consisting of haloC1-4alkyl, C1-4alkyl, OC1-4alkyl, OH, CN, and halogen, Wherein R5is selected from the group consisting of haloC1-4alkyl, C1-4alkyl, OC1-4alkyl, OH, CN, and halogen, wherein q is 0, 1, 2, 3 or 4; R6in each instance is independently selected from the group consisting of C1-4 alkyl, CN, halogen, phenyl, 5- or 6-membered heteroaryl, haloC1-4alkyl, C1-4alkyl, 3-6 membered- cycloalkyl, OC1-4 alkyl, O-haloC1-4 alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R6’in each instance is independently selected from the group consisting of C1-4alkyl, CN, halogen, phenyl, 5- or 6-membered heteroaryl, haloC1-4alkyl, C1-4alkyl, 3-6 membered- cycloalkyl, OC1-4 alkyl, O-haloC1-4 alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R6”is H or C1-4alkyl; a is 0, 1 or 2; b is 0, 1, 2, 3 or 4; r is is 0, 1, 2, 3 or 4; R7is selected from the group consisting of H, C1-4 alkyl, CN, and halogen, R8is H, or C1-4 alkyl, wherein s is 0, 1, 2, 3 or 4; R9is methyl or 3-6 membered-cycloalkyl, wherein R9is optionally substituted with one or more of C1-4 alkyl, CN, and halogen; R10in each instance is independently selected from the group consisting of C1-4 alkyl, CN, halogen, phenyl, 5- or 6-membered heteroaryl, haloC1-4alkyl, C1-4alkyl, 3-6 membered- cycloalkyl, OC1-4alkyl, O-haloC1-4alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R10’in each instance is independently selected from the group consisting of C1-4alkyl, CN, halogen, phenyl, 5- or 6-membered heteroaryl, haloC1-4alkyl, C1-4alkyl, 3-6 membered- 15 171560744182219.00268 cycloalkyl, OC1-4 alkyl, O-haloC1-4 alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R10”is H or C1-4alkyl; c is 0, 1 or 2; d is 0, 1, 2, 3 or 4; R11represents a substituent of the bicyclic ring it is attached to and in each instance is independently selected from the group consisting of C1-4alkyl, CN, halogen, phenyl, 5- or 6- membered heteroaryl, haloC1-4 alkyl, C1-4 alkyl, 3-6 membered-cycloalkyl, OC1-4 alkyl, O- haloC1-4alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R11’in each instance is independently selected from the group consisting of C1-4 alkyl, CN, halogen, phenyl, 5- or 6-membered heteroaryl, haloC1-4 alkyl, C1-4 alkyl, 3-6 membered- cycloalkyl, OC1-4alkyl, O-haloC1-4alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R11”is H or C1-4 alkyl; R12is H or a protecting group of the nitrogen that R12is attached to; e is 0, 1, 2, 3 or 4; f is 0, 1, 2, 3 or 4; R13represents a substituent of the bicyclic ring it is attached to and in each instance is independently selected from the group consisting of C1-4 alkyl, CN, and halogen; g is 0, 1, 2, or 3; t is 0, 1, 2, 3 or 4; u is 0, 1, 2, 3 or 4. In some embodiments, D-E together is one of the following: 16 171560744182219.00268, 17 171560744182219.00268, 18 171560744182219.00268.

[0068] In a compound containing one or more stereocenters, each can be independently R or S in configuration. In some embodiments, A is phenyl optionally substituted with one or more substituents selected from the group consisting of haloOC1-4alkyl, haloC1-4alkyl, C1-4alkyl, OC1-4 alkyl, OH, CN, halogen, NRaRb. In some embodiments, A is phenyl optionally substituted with haloOC1-4 alkyl, which can be at ortho, metal, or para position of the ring. In some embodiments, A is phenyl substituted with OCF3at para position. In some embodiments, B is amide.

[0069] In some embodiments, A is cyclopropyl optionally substituted with one or more substituents selected from the group consisting of haloOC1-4alkyl, haloC1-4alkyl, C1-4alkyl, OC1-4alkyl, OH, CN, halogen, and NRaRb. In some embodiments, A is cyclopropyl optionally substituted with phenyl, which can be optionally substituted with one or more of haloOC1-4 alkyl, haloC1-4alkyl, C1-4alkyl, OC1-4alkyl, OH, CN, halogen, and NRaRb. In some embodiments, A is cyclopropyl substituted with phenyl. In some embodiments, B is amide.

[0070] In some embodiments, A is phenyl substituted one or more substituents selected from the group consisting of O-haloC1-4 alkyl, haloC1-4 alkyl, C1-4 alkyl, CN, OC1-4 alkyl, O- haloC1-4alkyl, CN, and halogen.

[0071] In some embodiments, A is benzyl, which is optionally substituted with one or more of O-haloC1-4 alkyl, haloC1-4 alkyl, C1-4 alkyl, OC1-4 alkyl, OH, CN, halogen, and NRaRb. In some embodiments, B is amide.

[0072] In some embodiments, B forms a urea or an amide linkage with L1. When the amide linkage is formed, B is connected to A via the nitrogen of the amide or the C(O) of the amide. 19 171560744182219.00268

[0073] In some embodiments, B is amide, wherein each N can be optionally substituted with C1-4 alkyl. When a precursor of L1(e.g. alkylamino, piperidinyl or piperazinyl ring) contains a nitrogen for bonding with B, the nitrogen becomes part of a urea.

[0074] In some embodiments, B is amide which is linked to L1via a carbonyl (reverse amide).

[0075] In some embodiments, B is carbonyl. In some embodiments, A is benzyl, which is optionally substituted with one or more of haloC1-4alkyl, C1-4alkyl, CN, and halogen. In some embodiments, A is benzyl, substituted with CF3 and CN. In some embodiments, the CF3 is at ortho position and the CN is at para position. When a precursor of L1(e.g. alkylamino, piperidinyl or piperazinyl ring) contains a nitrogen for bonding with B, the nitrogen can become part of an amide.

[0076] In some embodiments, A-B are selected from the following. Each of the phenyl ring can be further substituted with one or more of haloC1-4alkyl, O-haloC1-4alkyl, C1-4alkyl, OC1-4 alkyl, CN, OH and halogen.

[0077] C serves to connect L1-L2with L3and is an amide, a carbonyl (C=O), or OC=O, or SO2. C as an amide can link up to L3via the nitrogen of the amide or the carbonyl of the amide (amide or reverse amide). Likewise, C can link up to L3via the oxygen of the OC=O or the carbonyl of the OC=O. The nitrogen of the amide, carbamate, or urea of C may be derived from a precursor of L1(if L2is void), L2or L3. For example, a precursor of L3when containing piperidinyl or piperazinyl as a component may have the nitrogen being part of the resulting carbamate, amide or urea of C. In some embodiments, C is SO2, which forms a sulfonamide with N of L2.

[0078] In some embodiments, C is C(O) and forms an amide linkage with L3.

[0079] In some embodiments, L1comprises a 3-6 membered ring selected from cycloalkyl, heterocycloalyl, phenyl or heteroaryl. In some embodiments, L1comprises cyclohexyl or 6 membered heterocycloalkyl. In some embodiments, L2is void. In some embodiments, L2comprises one or more of phenyl, C1-6 alkylene, O, and C(O). In some embodiments, L2comprises one or more of O-phenyl, C(O)-C1-4 alkylene, O-C1-4 alkylene- phenyl, and C1-4alkylene-phenyl. 20 171560744182219.00268

[0080] In some embodiments, L1comprises NH-3-6 membered cycloalkyl, or NH-3-6 membered heterocycloalkyl. In some embodiments, L2is void. In some embodiments, the NH of NH-3-6 membered cycloalkyl or NH-3-6 membered heterocycloalkyl is bonded to B.

[0081] In some embodiments, L1comprises NH-3-6 membered cycloalkyl or NH-3-6 membered heterocycloalkyl. In some embodiments, L2comprises one or more of phenyl, C1-6 alkylene, O, and C(O). In some embodiments, L2comprises or is phenyl, O-phenyl, C1-6alkylene, O, and C(O).

[0082] In some embodiments, L1comprises or consists of 3-6 membered cycloalkyl (no NH attached to the ring), 3-6 membered heterocycloalkyl, NH-3-6 membered cycloalkyl or NH-3-6 membered heterocycloalkyl, which links to B via its NH.

[0083] In some embodiments, L1comprises, or consists of, NH-3-6 membered cycloalkyl or NH-3-6 membered heterocycloalkyl, which is bonded to B via its NH.

[0084] The linkage between L1and L2can be ether (e.g. two rings linked via O) or amino (NH or N-alkyl).

[0085] In some embodiments, L2comprises or consists of O or NH bonded to an optionally substituted ring selected from the group consisting of phenyl, 5-6 membered cycloalkyl, 5-6 membered heterocycloalkyl, 5-6 membered heteroaryl.

[0086] In some embodiments, L2comprises O or NH bonded to an optionally substituted ring selected from the group consisting of phenyl and 6 membered heteroaryl (triazine, pyrimidine, pyridine, etc).

[0087] In some embodiments, L1-L2comprises or consists of NH-cyclohexyl-O-phenyl or cyclohexyl-NH-6-membered heteroaryl, wherein the phenyl and heteroaryl are optional substituted, wherein L1-L2is bonded to B via NH of NH- cyclohexyl -O-phenyl or cyclohexyl of cyclohexyl-NH-6-membered heteroaryl, wherein L2optionally further comprises one or more of C1-6 alkylene, C1-6 alkyleneC(O), and C1-6 alkyleneNH. 21 171560744182219.00268 In some embodiments, L1-L2comprises, which is bonded to B via the atom with a star. In some embodiments, C is bonded to L1-L2via a carbonyl.

[0088] Nonlimiting examples ofAinclude the following

[0089] In some embodiments, (L3)m comprises at least one of 3-6 membered cycloalkyl and 3-6 membered heterocycloalkyl. In some embodiments, (L3)mcomprises at least two rings of 3-6 membered cycloalkyl and / or a 3-6 membered heterocycloalkyl. L3can connect to any carbon of the D or E ring.

[0090] Nonlimiting examples of (L3)minclude the following, which can be connected to D or E at either end of the structure. 22 171560744182219.00268

[0091] In some embodiments, (L3)mis connected to D or E (when D is a bond) via ether, amino (NH or N-alkyl), amide (N bonded to D or E, or C(O) bonded to D or E), C2-15alkynlene, C1-15alkylene. In some embodiments, (L3)m includes one or more of an amide moietie, C1-10 alkylene, O-alkylene2-8NHC(O) (e.g.2-O-acetamide), [(CH2CH2)O]x(x=1-20), C(O)C1-10and alkylene.

[0092] When (L3)m is connected to D or E (when D is a bond) via O (ether), nonlimiting examples of components of (L3)minclude O-alkylene2-18,O-alkylene2-16NH, O-alkylene1-16C(O)NH, O-alkylene2-16NHC(O), (OCH2CH2)x(x=1-20), O-alkylene1-16C(O)NHalkylene1-16, O-alkylene1-16C(O)NH[(CH2CH2)O]x (x=1-20).

[0093] When (L3)m is connected to D or E (when D is a bond) via amino (NH or N- alkyl), nonlimiting examples of components of (L3)minclude NH-alkylene2-15,NH-alkylene2-23 171560744182219.00268 15NH, NH-alkylene2-15NHC(O), NH-alkylene2-15NHC(O) alkylene1-15, NH[(CH2CH2)O]x (x=1-20), NH[(CH2CH2)O]x alkylene2-15NH (x=1-20), NH[(CH2CH2)O]x alkylene2-15NHCO (x=1-20), NH[(CH2CH2)O]xalkylene2-15NH (x=1-20), NH[(CH2CH2)O]xalkylene2-15NHCO (x=1-20).

[0094] In some embodiments, (L3)m is connected to D or E (when D is a bond) via NHC(O), NHC(O)alkylene1-18, or C(O)NH C1-15alkylene. Either ends of these components can serve as the connection point. In some embodiments, the connection point is NH or C(O).

[0095] In some embodiments, (L3)m is connected to D or E (when D is a bond) via C2-15alkynleneNH, C2-15alkynleneNHC(O), or C2-15alkynleneNHC(O)C1-15alkylene. Either ends of these components can serve as the connection point. In some embodiments, the connection point is the alkynlene end.

[0096] In some embodiments, (L3)m is connected to D or E (when D is a bond) via C1-15alkyleneNH, or C1-15alkyleneNHC(O). Either ends of these components can serve as the connection point. In some embodiments, the connection point is the alkylene end.

[0097] In some embodiments, (L3)m is connected to D or E (when D is a bond) via C1-15alkyleneO, C1-15alkyleneOC1-15alkylene, C1-15alkyleneOC1-15alkyleneNHC(O), - C1-15alkyleneO[(CH2CH2)O]x(x=1-20), C1-15alkyleneO3-6 membered cycloalkyl, C1-15alkyleneO- 3-6 membered heterocycloalkyl, C1-15alkylene3-6 membered cycloalkyl, C1-15alkylene-3-6 membered heterocycloalkyl, C1-15alkyleneNH, C1-15alkyleneNHC(O), C1-15alkyleneC(O)NH, or C1-15alkylene-bicyclic. Either ends of these components can serve as the connection point. In some embodiments, the connection point is the alkylene end.

[0098] In some embodiments, (L3)mis connected to D or E (when D is a bond) via C(O)C1-15alkyleneO, C(O)C1-15alkyleneOC1-15alkylene, C(O)C1-15alkyleneOC1-15alkyleneNHC(O), C(O)C1-15alkyleneO[(CH2CH2)O]x (x=1-20), C(O) alkylene1-15 (OCH2CH2)x NHCO (x=1-20), C(O) alkylene1-15 (OCH2CH2)x (x=1-20), C(O)C1-15alkyleneO3- 6 membered cycloalkyl, C(O)C1-15alkyleneO-3-6 membered heterocycloalkyl, C(O)C1-15alkylene3-6 membered cycloalkyl, C(O)C1-15alkylene-3-6 membered heterocycloalkyl, C(O)C1-15alkyleneNH, C(O)C1-15alkyleneNHC(O), C(O)C1-15alkyleneC(O)NH, or C(O)C1-15alkylene-bicyclic. Either ends of these components can serve as the connection point. In some embodiments, the connection point is the C(O). C(O) may be part of an amide as the linkage between L3 and D or E. 24 171560744182219.00268

[0099] In some embodiments, (L3)m is connected to D or E (when D is a bond) via NHC(O)C1-15alkyleneO, NHC(O)C1-15alkyleneOC1-15alkylene, NH[(CH2CH2)O]x alkylene2-15NHCO (x=1-20), NHC(O)C1-15alkyleneOC1-15alkyleneNHC(O), NHC(O)C1-15alkyleneO[(CH2CH2)O]x (x=1-20), NHC(O)C1-15alkyleneO3-6 membered cycloalkyl, NHC(O)C1-15alkyleneO-3-6 membered heterocycloalkyl, NHC(O)C1-15alkylene3-6 membered cycloalkyl, NHC(O)C1-15alkylene-3-6 membered heterocycloalkyl, NHC(O)C1-15alkyleneNH, NHC(O)C1-15alkyleneNHC(O), NHC(O)C1-15alkyleneC(O)NH, or NHC(O)C1-15alkylene- bicyclic. Either ends of these components can serve as the connection point. In some embodiments, the connection point is the NH.

[0100] In some embodiments, (L3)mis connected to D or E (when D is a bond) via a phenyl, a 5-6 membered heteroaryl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkyl- alkylene1-15 cyclohexyl-alkylene1-15, cyclohexyl-alkylene1-15NHC(O), 3-6 membered cycloalkyl]1-4NHC(O), 3-6 membered cycloalkyl alkylene1-15NHC(O), 3-6 membered heterocycloalkyl -alkylene1-15-3-6 membered heterocycloalkyl, 3-6 membered cycloalkyl -alkylene1-15-3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkyl -alkylene1-15-3-6 membered cycloalkyl, or 3-6 membered cycloalkyl- alkylene1-15-3-6 membered cycloalkyl. Each of these rings is optionally substituted with one or more of haloOC1-4alkyl, haloC1-4 alkyl, C1-4 alkyl, 3-6 membered-cycloalkyl, OC1-4 alkyl, OH, CN, halogen, and NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; Either ends of these components can serve as the connection point. In some embodiments, the connection point is the ring end of these components.

[0101] In some embodiments, (L3)mis connected to D or E (when D is a bond) via C1-15alkylene3-6 membered heterocycloalkyl, C1-15alkylene3-6 membered heterocycloalkyl alkylene1-15NHC(O), alkylene1-15NHC(O), alkylene1-15NHC(O)alkylene1-15 , alkylene1-15O-3-6 membered heterocycloalkyl, alkylene1-15NH-3-6 membered heterocycloalkyl, alkylene1-15O- alkylene1-15-3-6 membered heterocycloalkyl, alkylene1-15NH-alkylene1-15-3-6 membered heterocycloalkyl, or alkylene1-15 - bicylic ring or tricyclic ring. Either ends of these components can serve as the connection point. In some embodiments, the connection point is alkylene ring end of these components.

[0102] In some embodiments, (L3)mis connected to D or E (when D is a bond) via a bicyclic ring or tricyclic ring (including bridged bicyclic or tricyclic, sprio-bicyclic or tricyclic), alkylene1-15-bicylic ring, or bicyclic ring-3-6 membered heterocycloalkyl. These components may include heteroatom in the ring. Either ends of these components can serve as 25 171560744182219.00268 the connection point. In some embodiments, the connection point is the ring end of these components.

[0103] In some embodiments, (L3)mis connected to D via a ring, C1-6alkylene, or C2-6alkynlene -3-6- membered cycloalkyl. In some embodiments, (L3)m is connected to D via C1-6 alkylene, C2-6 alkenlene or C2-6 alkynlene, which are further connected to 3-6- membered cycloalkyl or 3-6 membered heterocycloalkyl. In some embodiments, (L3)mcontains 1, 2 or 3 heterocyclalkyl ring. In some embodiments, (L3)mis connected to D via a ring such as 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl (e.g. piperidinyl, piperazinyl, cyclohexyl, pyrrolidinyl, etc, wherein the ring is optionally substituted with one or more substituents selected from haloC1-4alkyl, C1-4alkyl, OC1-4alkyl, OH, CN, and halogen. In some embodiments, a nitrogen atom of the ring is connected to D.

[0104] In some embodiments, (L3)m comprises one or more of the following. Preferably, (L3)mis bonded to D or E through the atom with a star. ,, .

[0105] In some embodiments, D is selected from the following and is connected to (L3)mat the phenyl moiety of D

[0106] In some embodiments, any of the D defined or illustrated in this patent document is optionally substituted with one or more of haloOC1-4 alkyl, haloC1-4 alkyl, C1-4 alkyl, CN, and halogen.

[0107] In some embodiments, D is selected from the group consisting of 26 171560744182219.00268bonded to E via the atom with a star, wherein D is optionally substituted with one or more of O-haloC1-4alkyl, haloC1-4alkyl, C1-4alkyl, CN, OC1-4alkyl, O-haloC1-4alkyl, CN, OH and halogen.

[0108] Nonlimiting examples of D-E include the following27 171560744182219.00268 ', wherein each ring atom of the above structures is optionally substituted with one or more of O-haloC1-4alkyl, haloC1-4alkyl, C1-4alkyl, CN, OC1-4alkyl, O-haloC1-4alkyl, CN, OH, and halogen.

[0109] In any embodiment disclosed herein where NH is present as in a amino group (e.g -NH-) or in an amide, the nitrogen is optionally substituted with a C1-6alkyl.

[0110] Nonlimiting examples of compounds of formula I Table 128 171560744182219.00268F3COOO N N N H N N N N HHO nO N O SF3F3F3F3F3F3F3F3F329 171560744182219.00268 O O O N O O H N 1-20 N NHF3C N N N N NOH H O H O O O O N O1-20OH NOF3FNNNN30 171560744182219.00268Further examples of the compound of Formula I include the following: ,, 31 171560744182219.00268 O O F O H O F O O O N F O N O O N N H H H OHN NOO O O O H O O O ONH O H H O N N N H ON O O O ON O OCF3O , O O O O N CF O3HN N N N N F O N O H H O , O OCF3O O N O HN N N N N F O N O H H , N O OCF O3O HN N N N N F O N O H H O , O CF N O3O HN N N N N O N O H H O , 32 171560744182219.00268 O O O O N HN OCF N N N O3O O N N N F OCF3F OCF3F33 171560744182219.00268.

[0111] Another aspect of the present disclosure provides a pharmaceutical composition containing a therapeutically effective amount of the above-described compound and a pharmaceutically acceptable carrier.

[0112] 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 composition disclosed 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.

[0113] 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 34 171560744182219.00268 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.

[0114] The pharmaceutical compositions may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, levigating, emulsifying, encapsulating, entrapping or tableting processes.

[0115] 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.

[0116] 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 thereof. Specific embodiments of the compound of Formula I are as described above.

[0117] The method disclosed herein can be applied to the treatment of various disease or disease conditions including, for example, inflammatory disease, cardiovascular disease, hypertension, myocardial infarction, ischemic stroke, neuropathic pain, depression, obesity, NAFLD, chronic obstructive pulmonary disease, diabetic retinopathy, cancer, metabolic dysfunction-associated steatohepatitis (mash), kidney dysfunction, gut barrier dysfunction, inflammatory pain, tooth regeneration, pulmonary hypertension, stroke, blood clotting, diabetic nephropathy, inflammatory pain, liver fibrosis, cardiovascular events, metabolic syndrome, cardiac hypertrophy, fibrosis, rheumatoid arthritis, hyperlipidemia, cardiovascular disease, neuroinflammation, stroke, neuropathic pain, pulmonary disease, renal disease, diabetes (type 1 and / or type 2), neurological disease, hypertension, pulmonary edema, pulmonary hypertension, cystic fibrosis, cardiomyopathy, hypertrophy of the heart, edema, pain, epilepsy, neuroma, cancer, Alzheimer's disease, dementia, amyotrophic lateral sclerosis (ALS), Parkinson's disease, prion diseases, depression, schizophrenia, and chemotherapy 35 171560744182219.00268 induced side effects. In varying embodiments, the subject may by symptomatic or asymptomatic for the disease or disease condition.

[0118] In some embodiments, the disease or disease condition is pain, including inflammatory pain and neuropathic pain. In varying embodiments co-administration of an agent that increases the production and / or level of epoxygenated fatty acids and an inhibitor of endoplasmic reticulum stress finds use in treating, i.e., reducing, relieving, ameliorating, mitigating, preventing, inhibiting and / or reversing neuropathic pain in a subject or patient in need thereof. The patient may be subject to suffering neuropathic pain chronically or intermittently. The patient may or may not be exhibiting or experiencing symptoms of neuropathic pain at the nine of treatment. The neuropathic pain may be centrally or peripherally mediated.

[0119] Neuropathic pain results from a pathology in the nervous system. Notable features of neuropathic pain include (1) widespread pain not otherwise explainable; (2) evidence of sensory deficit; (3) burning pain; (4) pain to light stroking of the skin (allodynia); and (5) enhanced stimulus-dependent pain (hyperalgesia) and (6) attacks of pain without seeming provocation (stimulus-independent pain). Neuropathic pain originates from a lesion of the nervous system (e.g., nerve damage). Any of a number of disease conditions or injuries can be the underlying cause of neuropathic pain. For example, the patient may be suffering from a metabolic disease (e.g., diabetic neuropathy), an autoimmune disease (e.g., multiple sclerosis), a viral infection (e.g. shingles and sequelae, postherpetic neuralgia), vascular disease (e.g. stroke), trauma and / or cancer. In varying embodiments, the neuropathic pain is due to nerve damage arising from one or more of trauma, ischemia or hemorrhage, inflammation, neurotoxicity, neurodegeneration, paraneoplastic, metabolic disease, vitamin deficiency, or cancer. In varying embodiments, the neuropathic pain is can be classified as toxic (e.g., arising from or secondary to chemoradiation or exposure to chemicals that cause nerve damage), metabolic (e.g., arising from or secondary to diabetes or nutritional deficiency, alcoholism), traumatic (e.g., arising from or secondary to phantom limb syndromes and / or complex regional pain syndromes (CRPS)), compressive (e.g., arising from or secondary to nerve entrapment and / or excessive external pressure on nerve axons which can cause ischemic or distortional (stretching) changes, or Wallerian degeneration of the axon with resultant muscle atrophy, autoimmune (e.g., arising from or secondary to autoimmune disease (e.g., Guillain-Barre Syndrome), chronic inflammatory demyelinating polyneuropathy (CIDP) and / or vasculitic neuropathy), infectious (e.g., arising from or secondary to an infectious disease, e.g., a viral 36 171560744182219.00268 infection such as Herpes Simplex Virus (HSV), Varicella Zoster Virus, Human Immunodeficiency Virus (HIV), a spirochete infection such as Lyme Disease, a trypanosome infection such as Chagas' Disease, a mycobacterium infection such as leprosy, and congenital / hereditary (e.g., arising from or secondary to Fabry's Disease, Charcot-Marie-Tooth Disease (burning pain in extremities), amyloidosis).

[0120] In some embodiments, the subject has been diagnosed peripheral neuropathic pain, for example, as a result of a disease condition including acute and chronic inflammatory demyelinating polyradiculoneuropathy; alcoholic polyneuropathy; chemotherapy-induced polyneuropathy; complex regional pain syndrome; entrapment neuropathies (e.g., carpal tunnel syndrome); HIV sensory neuropathy; iatrogenic neuralgias (e.g., postmastectomy pain or postthoracotomy pain); idiopathic sensor neuropathy; nerve compression or infiltration by tumor; nutritional deficiency-related neuropathies; painful diabetic neuropathy, phantom limb pain; postherpetic neuralgia; postradiation plexopathy; radiculopathy (cervical, thoracic, or lumbosacral); toxic exposure-related neuropathies; tic douloureux (trigeminal neuralgia); and / or posttraumatic neuralgias.

[0121] In some embodiments, the disease to be treated is characterized by alpha synuclein aggregates or deposits and is a neurodegenerative disease, e.g., selected from the group consisting of prodromal Parkinson's Disease, Parkinson's Disease, prodromal Dementia with Lewy Bodies (DLB), Dementia with Lewy Bodies (DLB), palsy, Pick's disease (frontal lobe dementia), and Alzheimer's Disease. In some embodiments, the mitigation comprises a reduction in the rate of alpha-synuclein aggregate or deposit formation in the brain of the subject. In some embodiments, the mitigation comprises a reduction in alpha-synuclein aggregate or deposit load in the brain of the subject. In some embodiments, the mitigation comprises an improvement in the cognitive and / or motor abilities of the subject. In some embodiments, the subject is a human. In some embodiments, the mitigation comprises a perceived improvement in quality of life by the human. In some embodiments, the subject is heterozygous or homozygous for a Parkinson's Disease associated mutation in one or more genes selected from the group consisting of alpha-synuclein (SNCA), leucine rich repeat kinase 2 (LRRK2), vacuolar protein sorting (VPS) retromer complex component (VPS35), parkin RBR E3 ubiquitin protein ligase (PRKN or PARK2), PTEN induced putative kinase 1 (PINK1) and glucocerebrosidase (GBA). In some embodiments, the subject has decreased expression of alpha-synuclein (SNCA) mRNA transcripts in circulating blood cells in comparison to a normal subject control. In some embodiments, the subject has decreased serum levels of heat 37 171560744182219.00268 shock protein family A (Hsp70) member 9 (HSPA9 or mortalin) in comparison to a normal subject control. In some embodiments, the subject has increased levels of monoamine oxidase B (MAOB), alpha-synuclein (SNCA) and / or soluble epoxide hydrolase (EPHX2) in lymphocytes, blood, serum or cerebrospinal fluid (CSF) in comparison to a normal subject control. In some embodiments, the blood alpha-synuclein (SNCA) of the subject has altered post-translational modifications, e.g., one or more of increased Y125 phosphorylation, increased Y39 nitration, and decreased SUMOylation, in comparison to a normal control subject. In some embodiments, the subject has decreased cerebrospinal fluid (CSF) levels of one or more biomarkers selected from the group consisting of Aβ1-42, T-tau, P-tau181, α- synuclein, and T-tau / Aβ1-42 in comparison to a normal control. In some embodiments, the subject has detectable nigrostriatal dopaminergic denervation, e.g., as detectable by single photon emission computed tomography (SPECT) or decarboxylase activity with 18F-Dopa positron emission tomography (PET). In some embodiments, the subject has decreased radiotracer binding using striatal dopamine transporters (DAT) / single photon emission computed tomography (SPECT) imaging. In some embodiments, the subject demonstrates subtle motor impairment. In some embodiments, the subject demonstrates mild cognitive impairment (MCI). In some embodiments, the subject demonstrates hyposmia, e.g., has failed an olfactory challenge test. In some embodiments, the subject suffers depression and / or anxiety.

[0122] In some embodiments, the subject has cardiomyopathy, cardiac arrhythmia or valvular heart disease. For example, the subject may have hypertrophic cardiomyopathy, e.g., due to valvular heart disease, familial hypertrophic cardiomyopathy, dilated cardiomyopathy, myocardial infarction, or secondary to administration of an anti-cancer drug or exposure to a toxic agent. Valvular heart disease can arise from any etiology, including, e.g., secondary to rheumatic fever, myxomatous degeneration of the valve, or papillary muscle dysfunction. In varying embodiments, the subject has cardiac arrhythmia, e.g., due to atrial fibrillation, ventricular fibrillation, or ventricular tachycardia.

[0123] In some embodiments, the subject is diagnosed with a disease where endoplasmic reticulum (ER) stress response is implicated in activating inflammation. Nonlimiting examples of the diseases include diabetes mellitus, obesity, non-alcoholic fatty liver disease (NAFLD), inflammatory bowel disease (IBD), atherosclerosis, neurodegenerative diseases (such as Alzheimer's disease, Parkinson's disease, and ALS), autoimmune diseases, and cancer. 38 171560744182219.00268

[0124] In some embodiments, the subject is diagnosed with a disease selected from diabetes, neuropathic pain, chronic obstructive pulmonary disease (COPD), metabolic disorders, Alzheimer's disease, Parkinson's disease, cardiovascular diseases, autism spectrum disorder (ASD) and schizophrenia, stroke, depression, alcohol associated liver disease, idiopathic pulmonary fibrosis (IPF), hypertension, asthma, diabetes, obesity, diabetic retinopathy, cardiac hypertrophy, pancreatic inflammation, pancreatic fibrosis, pulmonary fibrosis, renal fibrosis, chronic kidney disease, scarring, cisplatin toxicity, chemotherapy induced pain, atrial fibrillation, colitis, inflammatory bowel disease (IBD), sepsis (ARDS), metabolic dysfunction-associated steatohepatitis (MASH), osteoarthritis, rheumatoid arthritis, hardening of arteries, vascular inflammation, cardiac hypertrophy, atrial fibrosis, cardiac arrythmia, heat failure, myocardial infraction, traumatic brain injury, stroke, epilepsy, schizophrenia, depression, and pain (inflammatory and neuropathic).

[0125] In some embodiments, the subject is diagnosed with a disease selected from neurodegenerative diseases or disorders (e.g. Alzheimer's disease (AD), Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis and disorders caused by polyglutamine aggregation); skeletal muscle disease (e.g. Duchenne muscular dystrophy, skeletal muscle atrophy, Becker muscular dystrophy or myotonic Dystrophy); metabolic disorders (e.g. insulin resistance, diabetes, obesity, impaired glucose tolerance, high blood cholesterol, hyperglycemia, dyslipidemia and hyperlipidemia); adult-onset diabetes, diabetic nephropathy, neuropathy (e.g. sensory neuropathy, autonomic neuropathy, motor neuropathy, retinopathy); bone disease (e.g. osteoporosis), blood disease (e.g. leukemia); liver disease (e.g. due to alcohol abuse or hepatitis); Obesity; bone resorption, macular degeneration aging, AIDS-related dementia, ALS, Bell's palsy, atherosclerosis, heart disease (for example, arrhythmia, chronic congestive heart failure, ischemic stroke, coronary artery disease and cardiomyopathy), Chronic degenerative disease (e.g., myocardial disease), chronic renal failure, type 2 diabetes, ulcer, cataract, presbyopia, glomerulonephritis, Guillain-Barre syndrome, hemorrhagic stroke, rheumatoid arthritis, inflammatory bowel disease, SLE, Crohn's disease, Diseases or disorders associated with osteoarthritis, osteoporosis, chronic obstructive pulmonary disease (COPD), pneumonia, skin aging, urinary incontinence, mitochondrial dysfunction (e.g. mitochondrial myopathy, encephalopathy, Leber's disease, Lee encephalopathy, Pearson Disease, lactate acidosis, "mitochondrial encephalopathy, lactate acidosis and stroke-like symptoms" (MELAS), muscular diseases, including neuromuscular 39 171560744182219.00268 diseases, such as muscular dystrophy and myopathy, and diseases or disorders associated with neuronal death, aging, or other conditions characterized by unwanted cell loss.

[0126] In some embodiments, the methods disclosed herein further include administering to the subject an agent selected from the group consisting of celecoxib, pioglitazone, bardoxolone, sildenafil, pirfenidone, infliximab, semaglutide, Exenatide, Liraglutide, Dulaglutide, Lixisenatide, Efpeglenatide, Synergistic effect can be achieved when using a combination of agents. For instance, when used in combination with a COX-2 Inhibitor, dual targeting of prostaglandins and EET degradation can be achieved to suppress inflammation and angiogenesis synergistically.

[0127] Other agents that can be used as secondary agents include selective or nonselective COX Inhibitors (e.g. Aspirin, ibuprofen, naproxen, indomethacin, piroxicam, sulindac, diclofenac, ketoprofen, flurbiprofen, mefenamic acid, nabumetone, oxaprozin, tolmetin; Celecoxib, etoricoxib, parecoxib, valdecoxib, rofecoxib, lumiracoxib), PPARγ Activators (e.g. Rosiglitazone, pioglitazone, troglitazone, ciglitazone, rivoglitazone, balaglitazone, englitazone, netoglitazone, MCC-555, GW1929, LY-171883), Angiotensin Receptor Blockers (ARBs) (e.g. Losartan, valsartan, irbesartan, candesartan, olmesartan, telmisartan, eprosartan, azilsartan), Gabapentinoids / α2δ Ligands (e.g. Gabapentin, pregabalin, mirogabalin, phenibut, gabapentin enacarbil, 4-methylpregabalin, PD-217014), AMPK Activators (e.g. A769662, 991, PF-739, PF-06409577, MK-8722, ZLN024, C13, AICAR, metformin, salicylate, berberine, resveratrol), Nrf2 Activators (e.g., bardoxolone), Thrombolytics (e.g., tPA), NO Donors / PDE5 Inhibitors (e.g., sildenafil), Antifibrotics (e.g., pirfenidone), Anti-TNF Agents (e.g., infliximab), Omega-3 PUFA (e.g., EPA / DHA).

[0128] In some embodiments, the compound disclosed herein is used in combination with a GLP-1 Receptor Agonist. GLP-1RAs like semaglutide induce weight loss but also cause unintended muscle loss (sarcopenia), especially in aged or metabolically compromised individuals. sEH degraders enhance mitochondrial function, reduce ER stress, and preserve autophagic flux — all protective against muscle degeneration. sEH degradation may counteract GLP-1RA-induced sarcopenia. Combination therapy could preserve glucose-lowering and weight-loss effects of GLP-1RA while preventing or mitigating muscle atrophy. In some embodiments, the disease or condition is associated with downregulated or impaired GLP-1 (glucagon-like peptide-1). Nonlimiting examples include the aforementioned diseases. In some embodiments, the disease is diabetes (type 1 or 2) or obesity. 40 171560744182219.00268

[0129] Another aspect provides a method to increase survival rate of transplanted human induced pluripotent stem cell. The method includes treating the cell with an effective amount of the compound disclosed herein before transplantation. In some embodiments, the amount of the compound is selected so that the method improves the survival rate by at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 80%, or at least 90% in comparison with an untreated cell over the same period of time.

[0130] Another aspect provides a method of degrading soluble epoxide hydrolase (sEH). The method includes contacting the sEH with an effective amount of the compound of disclosed herein. In some embodiments, the amount of the compound is selected so that the method improves the survival rate by at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 80%, or at least 90% in comparison with an untreated cell over the same period of time.

[0131] The effective amount of compound or its pharmaceutical salt or composition is preferably in unit dosage form. In such form the compound or composition is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.

[0132] The term “unit dosage form”, as used in the specification, refers to physically discrete units suitable as unitary dosages for human subjects and animals, each unit containing a predetermined quantity of active material calculated to produce the desired pharmaceutical effect in association with the required pharmaceutical diluent, carrier or vehicle. The specifications for the novel unit dosage forms of this invention are dictated by and directly dependent on (a) the unique characteristics of the active material and the particular effect to be achieved and (b) the limitations inherent in the art of compounding such an active material for use in humans and animals, as disclosed in detail in this specification.

[0133] The dosage of the specific compounds depends on many factors that are well known to those skilled in the art. They include for example, the route of administration and the potency of the particular compound. An exemplary dose is from about 0.001 μg / kg to about 100 mg / kg body weight of the mammal. Determination of an effective amount is well within 41 171560744182219.00268 the capability of those skilled in the art, especially in light of the detailed disclosure provided herein. Generally, an efficacious or effective amount of a combination of one or more polypeptides of the present invention is determined by first administering a low dose or small amount of a polypeptide or composition and then incrementally increasing the administered dose or dosages, adding a second or third medication as needed, until a desired effect of is observed in the treated subject with minimal or no toxic side effects. Applicable methods for determining an appropriate dose and dosing schedule for administration of a combination of the present invention are described, for example, in Goodman and Gilman's The Pharmacological Basis of Therapeutics, 12th Edition, 2010, McGraw-Hill Professional; in a Physicians' Desk Reference (PDR), 68th Edition, 2014, PDR Network; in Remington: The Science and Practice of Pharmacy, 21st Ed., 2005, supra; and in Martindale: The Complete Drug Reference, Sweetman, 2005, London: Pharmaceutical Press., and in Martindale, Martindale: The Extra Pharmacopoeia, 31st Edition., 1996, Amer Pharmaceutical Assn, each of which are hereby incorporated herein by reference.

[0134] The compound of Formula I or a pharmaceutical composition thereof may also be used in combination with or include one or more other therapeutic agents, for example sEH inhibitors, Phosphodiesterase Inhibitors (PDEi), NSAIDS, corticosteroids, COX-2 inhibitors, cytokine inhibitors, anti-TNF agents, inhibitors oncostatin M, antimalarials, immunosuppressive and cytostatics. A number of other sEH inhibitors which can be used in the methods set forth in PCT / US2013 / 024396, PCT / US2012 / 025074, PCT / US2011 / 064474, PCT / US2011 / 022901, PCT / US2008 / 072199, PCT / US2007 / 006412, PCT / US2005 / 038282, PCT / US2005 / 08765, PCT / US2004 / 010298 and U.S. Published Patent Application Publication Nos: 2014 / 0088156, 2014 / 0038923, 2013 / 0274476, 2013 / 0143925, 2013 / 0137726, 2011 / 0098322, 2005 / 0026844, US2021016188, and US 10,813,894 each of which is hereby incorporated herein by reference in its entirety for all purposes. Other means of inhibiting sEH activity or gene expression can also be used in combination with the composition or the method disclosed herein. For example, a nucleic acid molecule complementary to at least a portion of the human sEH gene can be used to inhibit sEH gene expression. Means for inhibiting gene expression using short RNA molecules, for example, are known. Among these are short interfering RNA (siRNA), small temporal RNAs (stRNAs), and micro-RNAs (miRNAs). Short interfering RNAs silence genes through a mRNA degradation pathway, while stRNAs and miRNAs are approximately 21 or 22 nt RNAs that are processed from endogenously encoded hairpin-structured precursors, and function to silence genes via translational repression. 42 171560744182219.00268

[0135] In some embodiments, the compound or pharmaceutical salt thereof disclosed herein is administered in conjunction with or concurrently with one or more agents selected from the group consisting of an inhibitor of cyclooxygenase-2 (COX-2), and inhibitor of phosphodiesterase, and mixtures thereof. In some embodiments, the sEHI is administered in conjunction with an agent selected from the group consisting of inhibitor of endoplasmic reticulum (ER) stress, carbidopa / levodopa, dopamine agonists, and monoamine oxidase type B (MAO-B) inhibitors, and mixtures thereof. In some embodiments, the inhibitor of ER stress is selected from the group consisting of group consisting of 4-phenyl butyric acid (4-PBA), 3- phenylpropionic acid (3-PPA), 5-phenylvaleric acid (5-PVA), 6 phenylhexanoic acid (6-PHA), butyrate, tauroursodeoxycholic acid, trehalose, deuterated water, docosahexaenoic acid (“DHA”), eicosapentaenoic acid (“EPA”), vitamin C, arabitol, mannose, glycerol, betaine, sarcosine, trimethylamine-N oxide, DMSO and mixtures thereof.

[0136] In some embodiments, the compound or pharmaceutical salt thereof disclosed herein is co-administered with an enhancing or synergizing agent. In some embodiments, the secondary agents include without limitation inhibitors of cyclooxygenase-2 (COX-2), inhibitors of phosphodiesterase, agonists of peroxisome proliferator activated receptor alpha (PPARα) and agonists of peroxisome proliferator activated receptor gamma (PPARγ).

[0137] Further examples of inhibitors of COX-2 that may be co-administered with the compound or pharmaceutical salt thereof disclosed herein include without limitation celecoxib, valdecoxib, lumiracoxib, etoricoxib, and rofecoxib. Illustrative inhibitors of phosphodiesterase 4 that may be co-administered with an inhibitor of soluble epoxide hydrolase include without limitation rolipram, roflumilast, cilomilast, ariflo, HT0712, ibudilast and mesembrine. Illustrative inhibitors of phosphodiesterase 5 that may be co-administered with an inhibitor of soluble epoxide hydrolase include without limitation sildenafil, zaprinast, tadalafil, udenafil, avanafil and vardenafil. Illustrative agonists of PPARα that may be co- administered with an inhibitor of soluble epoxide hydrolase include without limitation clofibrate, gemfibrozil, ciprofibrate, bezafibrate, and fenofibrate. Illustrative agonists of PPARγ that may be co-administered with an inhibitor of soluble epoxide hydrolase include without limitation thiazolidinediones (TZDs).

[0138] 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 and mammalian species treated, the particular compounds employed, and the specific use for which these compounds are employed. The determination of effective dosage levels, 43 171560744182219.00268 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.

[0139] 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.

[0140] 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.

[0141] 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 44 171560744182219.00268 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.

[0142] In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.

[0143] 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.

[0144] 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 such studies 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.

[0145] 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 45 171560744182219.00268 prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0146] 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.

[0147] Another aspect of the patent document provides a method of degrading a sEH in a cell. The method includes contacting the cell with an effective amount of a compound of Formula I disclosed herein, wherein the compound effectuates the degradation of the target protein in the cell. In some embodiments, the cell is in a human.

[0148] Another aspect of the patent document provides a method of inhibiting sEH in a cell. The method includes contacting the cell with an effective amount of a compound of Formula I disclosed herein, wherein the compound effectuates the degradation of the target protein in the cell. In some embodiments, the cell is in a human.

[0149] Examples

[0150] Example 1. Compound synthesis

[0151] Synthesis of ALT-PG2, compounds 1, 2, and s5. Compound s1 (10.5 mg, 24.0 μmol, 1.0 equiv.) and HTAU (18.3 mg, 48.0 mmol, 2.0 equiv.) were added to a solution of amine derivative (24.0 μmol, 1.0 equiv.) in DMF (500 μL) and DIEPA (50 μL, 287 μmol, 12.0 equiv.). The reaction mixture was stirred at room temperature for 15 hours, then filtered through a PTFE membrane and purified via preparative HPLC using H2O-MeCN gradient (99: 1 to 5: 95, v / v, 0.1% FA). A fraction containing the target molecule was lyophilized to give a solid. 46 171560744182219.00268

[0152] Synthesis of compound s2. Compound s1 (90.4 mg, 206 μmol, 1.0 equiv.) and HTAU (158.6 mg, 417 μmol, 2.0 equiv.) were added to a solution of tert-butyl (14-amino- 3,6,9,12-tetraoxatetradecyl)carbamate (from AmBeed, 90.2 mg, 268 μmol, 1.3 equiv.) in DMF (2.0 mL) and DIEPA (180 μL, 1.03 mmol, 5.0 equiv.). The reaction mixture was stirred at room temperature for 15 hours, then extracted with ethyl acetate three times. The combined organic layer was dried over Na2SO2, filtered, and concentrated in vacuo. The residue was further purified by flash chromatography (hexane: ethyl acetate).

[0153] Synthesis of compounds 3, 4, and 5. Compound s2 (10.0 mg, 13.2 μmol, 1.0 equiv.) was dissolved in TFA (1.0 mL) and the reaction mixture was stirred at room temperature for 1 hour, then concentrated in vacuo. The residue and HATU (10.0 mg, 26.4 μmol, 2.0 equiv.) were added to a solution of carboxylate derivative (13.2 μmol, 1.0 equiv.) in DMF (500 mL) and DIEPA (20 μL, 115 μmol, 8.7 equiv.). The reaction mixture was stirred at room temperature for 15 hours, then filtered through a PTFE membrane and purified via preparative HPLC using a H2O-MeCN gradient (99: 1 to 5: 95, v / v, 0.1% FA). A fraction containing the target molecule was lyophilized to give a solid. 47 171560744182219.00268

[0154] Compound s2 (10.0 mg, 13.2 μmol, 1.0 equiv.) was dissolved in TFA (1.0 mL) and the reaction mixture was stirred at room temperature for 1 hour, then concentrated in vacuo. The residue and HATU (10.0 mg, 26.4 μmol, 2.0 equiv.) were added to a solution of carboxylate derivative (13.2 μmol, 1.0 equiv.) in DMF (500 mL) and DIEPA (20 μL, 115 μmol, 8.7 equiv.). The reaction mixture was stirred at room temperature for 15 hours, then filtered through a PTFE membrane and purified via preparative HPLC using a H2O-MeCN gradient (99: 1 to 5: 95, v / v, 0.1% FA). A fraction containing the target molecule was lyophilized to give a solid.

[0155] Synthesis of compound s3. Compound s1 (140.8 mg, 321 μmol, 1.0 equiv.) and HTAU (244.2 mg, 642 mmol, 2.0 equiv.) were added to a solution of tert-butyl (14-amino- 3,6,9,12-tetraoxatetradecyl)carbamate (from AmBeed, 110.5 mg, 482 μmol, 1.5 equiv.) in DMF (2.0 mL) and DIEPA (300 μL, 1.72 mmol, 5.4 equiv.). The reaction mixture was stirred at room temperature for 15 hours, then extracted with ethyl acetate three times. The combined organic layer was dried over Na2SO2, filtered, and concentrated in vacuo. The residue was further purified by flash chromatography (hexane: ethyl acetate). 48 171560744182219.00268

[0156] Synthesis of compound s4

[0157] Compound s4 was prepared by the similar procedure as that described in literature.4-(Trifluoromethoxy)phenyl isocyanate (150 μL, 994 μmol, 1.0 equiv.) and 4-amino- 1-Boc-piperidine (239.5 mg, 1.20 mmol, 1.2 equiv.) was dissolved in CH2Cl2 (10 mL) and stirred for 2 hours at room temperature. The reaction was quenched by addition of water (5 mL) and then the solution was extracted with ethyl acetate three times. The combined organic layer was dried over Na2SO4, filtered, and concentrated in vacuo. The residue was further purified by flash chromatography (hexane: ethyl acetate).

[0158] Synthesis of compounds 9, 10, and 11

[0159] Compound s4 (10.0 mg, 24.8 μmol, 1.0 equiv.) was dissolved in TFA (1.0 mL) and the reaction mixture was stirred at room temperature for 1 hour, then concentrated in vacuo. The residue and HATU (18.9 mg, 49.6 μmol, 2.0 equiv.) were added to a solution of carboxylate derivative (24.8 μmol, 1.0 equiv.) in DMF (500 μL) and DIEPA (40 μL mg, 230 μmol, 9.3 equiv.). The reaction mixture was stirred at room temperature for 15 hours, then filtered through a PTFE membrane and purified via preparative HPLC using a H2O-MeCN gradient (99: 1 to 5: 95, v / v, 0.1% FA). A fraction containing the target molecule was lyophilized to give a solid. 49 171560744182219.00268

[0160] Synthesis of compounds 12 and 13. Compound s4 (10.0 mg, 24.8 μmol, 1.0 equiv.) was dissolved in TFA (1.0 mL) and the reaction mixture was stirred at room temperature for 1 hour, then concentrated in vacuo. The residue and HATU (18.9 mg, 49.6 μmol, 2.0 equiv.) were added to a solution of carboxylate derivative (24.8 μmol, 1.0 equiv.) in DMF (500 μL) and DIEPA (40 μL mg, 230 μmol, 9.3 equiv.). The reaction mixture was stirred at room temperature for 15 hours, then filtered through a PTFE membrane and purified via preparative HPLC using a H2O-MeCN gradient (99: 1 to 5: 95, v / v, 0.1% FA). A fraction containing the target molecule was lyophilized to give a solid.

[0161] Synthesis of compound s6. Compound s6 (115.7 mg, 286.8 μmol, 1.0 equiv.) was dissolved in TFA (200 μL) and CH2Cl2 (2 mL), and the reaction mixture was stirred at room temperature for 1 hour, then concentrated in vacuo. The residue and HATU (172.26 mg, 453.0 μmol, 1.6 equiv.) were added to a solution of 10-(tert-butoxy)-10-oxodecanoic acid (from AmBeed, 125.3 mg, 484.8 μmol, 1.7 equiv.) in DMF (3.0 mL) and DIEPA (200 μL, 1.15 mmol, 4.0 equiv.). The reaction mixture was stirred at room temperature for 15 hours, then extracted with ethyl acetate three times. The combined organic layer was dried over Na2SO4, filtered, and concentrated in vacuo. The residue was further purified by flash chromatography (hexane: ethyl acetate).

[0162] Synthesis of compound s6 50 171560744182219.00268

[0163] Compound s6 (115.7 mg, 286.8 μmol, 1.0 equiv.) was dissolved in TFA (200 μL) and CH2Cl2 (2 mL), and the reaction mixture was stirred at room temperature for 1 hour, then concentrated in vacuo. The residue and HATU (172.26 mg, 453.0 μmol, 1.6 equiv.) were added to a solution of 10-(tert-butoxy)-10-oxodecanoic acid (from AmBeed, 125.3 mg, 484.8 μmol, 1.7 equiv.) in DMF (3.0 mL) and DIEPA (200 μL, 1.15 mmol, 4.0 equiv.). The reaction mixture was stirred at room temperature for 15 hours, then extracted with ethyl acetate three times. The combined organic layer was dried over Na2SO4, filtered, and concentrated in vacuo. The residue was further purified by flash chromatography (hexane: ethyl acetate).

[0164] Synthesis of compound 19

[0165] Compound s6 (9.6 mg, 17.6 μmol, 1.0 equiv.) was dissolved in TFA (800 μL) and the reaction mixture was stirred at room temperature for 1 hour, then concentrated in vacuo. The residue and HATU (13.4 mg, 35.2 mmol, 2.0 equiv.) were added to a solution of pomalidomide 5'-piperazine-4-methylpiperidine (from Tocris, 7.7 mg, 17.6 μmol, 1.0 equiv.) in DMF (500 μL) and DIEPA (20 μL, 110 μmol, 6.5 equiv.). The reaction mixture was stirred at room temperature for 15 hours, then filtered through a PTFE membrane and purified via preparative HPLC using a H2O-MeCN gradient (99:1 to 5:95, v / v, 0.1% FA). A fraction containing the target molecule was lyophilized to give a solid.

[0166] Example 2

[0167] In the course of evaluating the cellular activity of the first-generation sEH PROTAC molecules, compounds 1a and ALT-PG2 were found to be almost completely degraded in the cell culture medium (DMEM) after 24 hours of incubation. It was hypothesized that this is due to the hydrolysis of the O-linked thalidomide moiety. A range of cereblon- recruiting fragments and linkers were selected that were previously reported to exhibit greater aqueous stability than the O-linked thalidomide moiety. In addition to t-TUCB (4-[[trans-4- [[[[4-(trifluoromethoxy)phenyl]amino]carbonyl]amino]cyclohexyl]oxy]benzoic acid) scaffold 51 171560744182219.00268 that was used as an sEH binding moiety for the development of 1a and ALT-PG2, TPPU (1- (1-propanoylpiperidin-4-yl)-3-[4-(trifluoromethoxy)phenyl]urea) scaffold was also used to determine the influence of sEH binding moiety on the degradation potency and stability. X-ray structures of these molecules bound to human sEH indicated that portions of the molecules are exposed to the solvent, making them amenable to modifications. Eight new PROTAC candidate molecules were designed based on t-TUCB and five molecules based on TPPU. Briefly, E3 ligase recruiter fragments, linkers and sEH binding moieties were connected by using amide coupling. TPPU scaffold was synthesized by reacting the corresponding isocyanate with Boc-protected 4-aminopiperidine.

[0168] The cellular degradation potency of the synthesized molecules was first evaluated at a single concentration (1 µM) on HepG2 cells. ELISA was used for sEH quantification given its throughput and quantitative nature. As shown in Tables 2 and 3, subtle changes in the linker and E3 recruiter structures appear to influence the degradation potency of the molecules. Importantly, the t-TUCB series molecules with a piperazine linker (compounds 6 - 8) demonstrated superior sEH degradation compared to the original PROTAC 1a. These piperazine linkers and E3 recruiters have been employed in several of the first PROTACs that are moving into the clinical trials, such as ARV-110 and ARV471. Phenyl dihydrouracil and phenyl glutarimide were reported to be alternative cereblon binders with improved stability. Unfortunately, in this case, compounds with these fragments did not show significant degradation of sEH (compounds 3 and 4). Comparison between compounds 4 and 5 indicate that the linker position on phenyl dihydrouracil is important for the degradation. When comparing t-TUCB series and TPPU series with the same linker / E3 recruiter moiety (compound 1 vs. 11), TPPU-based compound 11 showed slightly better degradation than t- TUCB-based molecule 1, indicating the potential of this scaffold for further optimization. In addition to the cellular sEH degradation, the biochemical enzyme inhibitory potency was measured against both human and mouse sEH using a fluorogenic substrate. All molecules showed sEH inhibitory activity ranging from sub-nanomolar to low nanomolar potency.

[0169] Synthesis of t-TUCB-based sEH PROTACs. a) HATU and DIEPA in DMF, b) 1) trifluoroacetic acid, RT.2) HATU and DIEPA in DMF, RT. 52 171560744182219.00268

[0170] Table 2. Structure of t-TUCB-based sEH PROTAC molecules and their cellular degradation and biochemical inhibitory potency against sEH53 171560744182219.00268aCellular sEH degradation was measured in HepG2 cells at 1 µM for 24 hours and the values are normalized based on the total protein concentration and DMSO treated samples in triplicates.bBiochemical inhibitory potency was measured against recombinant purified human and mouse sEH using a fluorescent substrate for hydrolase activities. Reported IC50 values are the average of triplicates with at least two data points above and at least two below the IC50. The fluorescent-based assay has a standard error between 10% and 20%, suggesting that differences of 2-fold or greater are significant.

[0171] Table 3. Structure of TPPU-based sEH PROTAC molecules and their degradation and inhibitory potency against sEHaCellular sEH degradation was measured in HepG2 cells at 1 µM for 24 hours and the values are normalized based on the total protein concentration and DMSO treated samples in triplicates. 54 171560744182219.00268bBiochemical inhibitory potency was measured against recombinant purified human and mouse sEH using a fluorescent substrate for hydrolase activities. Reported IC50 values are the average of triplicates with at least two data points above and at least two below the IC50. The fluorescent-based assay has a standard error between 10% and 20%, suggesting that differences of 2-fold or greater are significant.

[0172] Encouraged by the initial structure-degradation relationship results, we chose molecules 6 and 8 were chosen for further characterization. First, we measured the aqueous stability of compounds 6 and 8 was measured in the cell culture medium (DMEM). These molecules showed much higher stability compared to compound 1a in the medium. In addition, the in vitro metabolic stability of the compounds was assessed against mouse liver microsome. Both 8 (T1 / 2>120 min) and 6 (T1 / 2= 31 min) were found to be much more stable compared to 1a (T1 / 2= 0.59 min) (Table 4).

[0173] Table 4. Aqueous and in vitro metabolic stability of sEH PROTACsaStability in cell culture media was measured in DMEM.bMicrosomal stability was measured by incubating compound at 1 µM with mouse liver microsome (0.5 mg / mL) and NADPH (2 mM) solution for 0 - 45 minutes in duplicates.cValues measured without NADPH with 45 minutes incubation.

[0174] To evaluate their precise degradation potency, a dose-dependent sEH degradation induced by PROTACs 8 and 6 was next determined. Compound 8 was found to have higher degrading potency compared to compound 6. Remarkably, compound 8 induced degradation with a half-maximal degradation concentration (DC50) around 0.5 nM. A dose- response with wider concentration range (0.01 - 1000 nM) of sEH PROTAC 8 showed that maximum degradation is around 100 nM with a maximal level of degradation (Dmax) at 79%.

[0175] The improvement of degradation potency was further assessed by direct comparison of the most promising molecule 8 with the first generation PROTACs 1a and ALT- PG2 at two different concentrations. The results aligned with the ELISA data, demonstrating that compound 8 induce a significantly higher degradation of sEH than compound 1a and ALT- PG2. Both 1a and ALT-PG2 failed to induce 50% degradation at 250 nM and 1 μM (100 nM - 1 µM; DC50 > 1 µM), while compound 8 robustly degraded sEH more than 50% at both 55 171560744182219.00268 concentrations. Based on these data, the new generation PROTAC 8 has 500 - 2000-fold improvements in DC50 with higher Dmax compared to the PROTACs previously reported. Taken together, compound 8 has significantly improved degradation potency against sEH with improved metabolic and aqueous stability.

[0176] Compound 8 was selected for further biological evaluations. The degradation kinetics of sEH induced by compound 8 was first determined. HepG2 cells were treated with 50 nM of 8, and the amount of sEH was measured at various incubation time using immunoblotting. Compound 8 showed degradation of more than 50% of sEH as fast as 4 hours, with the highest level of degradation after 24 hours of incubation. The degradation kinetics is much faster than the first generation PROTAC 1a that required 16 hours to show sufficient degradation of sEH.

[0177] The mechanisms of degradation was next validated by co-treatment of compound 8 (50 nM) with sEH inhibitor t-TUCB (1 µM), lenalidomide (1 µM, E3 ligase ligand), MLN4924 (1 µM, an inhibitor of NEDD8-activating enzyme), or MG-132 (10 or 40 µM, proteasome inhibitor). All of these co-treatments inhibited sEH degradation induced by compound 8, supporting the mechanism of degradation of PROTACs. The subcellular selective sEH degradation was determined in cytosol and peroxisome. Aligned with previous results of compound 1a, the new compound 8 also induced cytosol selective sEH degradation over peroxisomal sEH. Finally, quantitative MS-based proteomic analysis revealed that sEH is among the proteins most significantly reduced following treatment with 8.

[0178] Prior to progressing with animal testing of our compounds, an additional cycle of medicinal chemistry was conducted to broaden the range of candidate molecules and to enhance our understanding of the structure-degradation relationships. Six additional sEH PROTAC candidate molecules were designed based on the structures of the 8 and 6 (Table 4). The sEH degradation activity of the synthesized molecules was evaluated by both immunoblotting and ELISA in parallel this time. It was found that compounds 17, 19, and 18 have degradation potency comparable to 8. Compound 18, an analog of 6 with lenalidomide 5’-piperazine fragment as an E3 ligase recruiter, was found to be relatively unstable in the medium, with 57% remaining after 24 hours of incubation. Moreover, this compound showed cytotoxicity against HepG2 at 1 µM. On the other hand, compound 17, a structural analog of compound 8 with lenalidomide 5’-piperazine-4-methylpiperidine fragment as an E3 ligase recruiter, showed high sEH degradation potency with stability in the cell culture medium and no cytotoxic effects. Compound 19, which has TPPU scaffold as a sEH binder with the E3 ligase recruiter and linker used in the compound 8, was found to have high degradation potency, 56 171560744182219.00268 while it was found to be less chemically stable than compound 8. Molecules with different linker positions (compounds 14 and 15) induced less degradation than the corresponding molecules that has the linker at 5 position of pomalidomide, indicating the importance of linker position for degradation potency.

[0179] Compounds 17 and 19 with high sEH-degrading activity and reasonable aqueous stability were studied. A dose-response experiment of sEH PROTACs 17 and 19 (0.03 - 10 nM) in HepG2 cells showed that compound 17 has degradation potency comparable to compound 8, while compound 19 showed slightly less potency. A potential reason for the lower activity of compound 19 compared to compound 8 may be due to the lower stability in the cell culture medium. In addition, these molecules along with compound 8 effectively degraded sEH in HEK293T, and compound 8 also degraded sEH in human breast adenocarcinoma MDA- MB-231 cells and human bone marrow neuroblast SH-SY5Y cells. Overall, among the compounds synthesized, compounds 8 and 17 were found to have the highest sEH-degrading activity, followed by compound 19. Importantly, both compounds 8 and 19 have the same linker but different E3 ligase recruiting moieties (pomalidomide vs. lenalidomide), indicating the importance of the linker for the sEH degradation.

[0180] Table 4. Structure-degradation relationships of compound 8 analogs57 171560744182219.00268aCellular sEH degradation was measured in HepG2 cells for 24 hours and the values are normalized based on the total protein concentration and DMSO treated samples in triplicates. Raw data and SD values are shown in.bCytotoxicity was measured using HepG2 cells treated with 1 µM compound for 24 hours. MTT assay was used to measure cell viability.cBiochemical inhibitory potency was measured against recombinant purified human and mouse sEH using a fluorescent substrate for hydrolase activities. Reported IC50 values are the average of triplicates with at least two data points above and at least two below the IC50. The fluorescent-based assay has a standard error between 10% and 20%, suggesting that differences of 2-fold or greater are significant.

[0181] Pharmacokinetics and sEH degradation in vivo. Based on the degradation potency and chemical stability, compounds 8, 17, and 19 were selected for in vivo pharmacokinetic (PK) profiling. Compounds were administered to male CD1 mice (weighting 30 - 31 g, n = 3) in a single i.p. injection (10 mg / kg). Plasma concentration of the molecules was monitored at various time points. Compound 8 was found to be most stable with half-life at 12 hours, followed by compound 17 with half-life around 7 hours. TPPU series compound 19 was found to be least stable in vivo with limited AUC values. The plasma concentration of compound 8 was maintained above 1 µM (>1,000-fold higher than DC50) over the course of PK profiling even after 24 hours of single i.p. injection.

[0182] Finally, the effect of compound 8 on sEH degradation in vivo was evaluated. Mice (male C57BL, weighting 22 - 28 g, n = 2) were treated with compound 8 (12 or 36 mg / kg, single i.p. injection). After 24 hours, mice were sacrificed and the sEH level in the liver and 58 171560744182219.00268 brown adipose tissue (BAT) was measured using immunoblotting. Strikingly, a significant degradation of sEH was observed in both liver and BAT, with higher degradation in the liver.

[0183] 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. 59 171560744

Claims

182219.00268 WE CLAIM 1. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is represented by Formula I:Formula I wherein A is a C3-6cycloalkyl, a phenyl, 5- or 6-membered heteroaryl, benzyl or, wherein A is optionally substituted with one or more substituents selected from the group consisting of O- haloC1-4alkyl, phenyl, 5- or 6-membered heteroaryl, haloC1-4alkyl, C1-4alkyl, 3-6 membered- cycloalkyl, OC1-4alkyl, O-haloC1-4alkyl, OH, CN, halogen, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; B is a carbonyl (C=O) or an amide, wherein the carbonyl carbon of the amide is bonded to L1; C is an amide, a carbonyl (C=O), or OC=O, wherein the carbonyl carbon of the amide or the OC=O is bonded to (L3)m; D is void or a bicyclic ring or a single aromatic ring, wherein the ring is optionally substituted with one or more substituents selected from the group consisting of haloOC1-4alkyl, haloC1-4alkyl, C1-4 alkyl, 3-6 membered-cycloalkyl, OC1-4 alkyl, OH, CN, halogen, NRcRd, wherein Rcand Rdin each instance is independently H or C1-4alkyl;optionally substituted with one or more substituents selected from the group consisting of O-haloC1-4alkyl, haloC1-4alkyl, C1-4alkyl, 3-6 membered-cycloalkyl, OC1-4 alkyl, OH, CN, halogen, NRcRd, wherein Rcand Rdin each instance is independently H or C1-4 alkyl; L1is a linker comprising 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, NH- 3- 6 membered cycloalkyl, or NH-3-6 membered heterocycloalkyl; 60 171560744182219.00268 L2is a bond or a linker comprising one or more of phenyl, 5-6 membered heteroaryl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, C1-6alkylene, O, C(O), NH, and NC1-6alkyl, wherein the phenyl, 5-6 membered heteroaryl, 3-6 membered cycloalkyl, and 3-6 membered heterocycloalkyl are each optionally substituted with one or more of O-haloC1-4alkyl, haloC1-4alkyl, C1-4alkyl, CN, and halogen; L3is a bond or a linker comprising one or more of phenyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, bicyclic ring, tricyclic ring, C1-6 alkylene, C2-6 alkenlene, C2-6 alkynlene, O, C(O), NH, (OCH2CH2)1-20and NC1-6alkyl; and m is an integer from 1 to 20; provided that the compound is not the following:alternatively, A-B wherein R1and R2in each instance are independently selected from the group consisting of haloOC1-4 alkyl, haloC1-4 alkyl, C1-4 alkyl, OC1-4alkyl, OH, CN, halogen, NReRf, wherein Reand Rfin each instance is independently H or C1-4alkyl, and L1-L2is a linker comprising C1-8alkylene, 3-6 membered 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl; n and p are independently 0, 1, 2, or 3; alternatively, D-E together is selected from the group consisting of 61 171560744182219.00268171560744182219.00268Wherein R3is selected from the group consisting of H, C1-4alkyl, and C(O)C1-4alkyl, R4is selected from the group consisting of H, phenyl and benzyl, wherein the phenyl and benzyl is optionally substituted with one or more substituents selected from the group consisting of haloC1-4alkyl, C1-4alkyl, OC1-4alkyl, OH, CN, and halogen, R5is selected from the group consisting of haloC1-4 alkyl, C1-4 alkyl, OC1-4 alkyl, OH, CN, and halogen, wherein q is 0, 1, 2, 3 or 4; R6in each instance is independently selected from the group consisting of C1-4alkyl, CN, halogen, phenyl, 5- or 6-membered heteroaryl, haloC1-4 alkyl, C1-4 alkyl, 3-6 membered- cycloalkyl, OC1-4 alkyl, O-haloC1-4 alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R6’in each instance is independently selected from the group consisting of C1-4alkyl, CN, halogen, phenyl, 5- or 6-membered heteroaryl, haloC1-4 alkyl, C1-4 alkyl, 3-6 membered- cycloalkyl, OC1-4alkyl, O-haloC1-4alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R6”is H or C1-4 alkyl; a is 0, 1 or 2; b is 0, 1, 2, 3 or 4; R9is methyl or 3-6 membered-cycloalkyl, wherein R9is optionally substituted with one or more of C1-4 alkyl, CN, and halogen; R10in each instance is independently selected from the group consisting of C1-4alkyl, CN, halogen, phenyl, 5- or 6-membered heteroaryl, haloC1-4alkyl, C1-4alkyl, 3-6 membered- 63 171560744182219.00268 cycloalkyl, OC1-4alkyl, O-haloC1-4alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R10’in each instance is independently selected from the group consisting of C1-4 alkyl, CN, halogen, phenyl, 5- or 6-membered heteroaryl, haloC1-4alkyl, C1-4alkyl, 3-6 membered- cycloalkyl, OC1-4alkyl, O-haloC1-4alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R10”is H or C1-4 alkyl; c is 0, 1 or 2; d is 0, 1, 2, 3 or 4; R11represents a substituent of the bicyclic ring it is attached to and in each instance is independently selected from the group consisting of C1-4alkyl, CN, halogen, phenyl, 5- or 6- membered heteroaryl, haloC1-4alkyl, C1-4alkyl, 3-6 membered-cycloalkyl, OC1-4alkyl, O- haloC1-4 alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R11’in each instance is independently selected from the group consisting of C1-4alkyl, CN, halogen, phenyl, 5- or 6-membered heteroaryl, haloC1-4 alkyl, C1-4 alkyl, 3-6 membered- cycloalkyl, OC1-4 alkyl, O-haloC1-4 alkyl, OH, NRaRb, wherein Raand Rbin each instance is independently H or C1-4alkyl; R11”is H or C1-4 alkyl; R12is H or a protecting group of the nitrogen that R12is attached to; e is 0, 1, 2, 3 or 4; f is 0, 1, 2, 3 or 4; R13represents a substituent of the bicyclic ring it is attached to and in each instance is independently selected from the group consisting of C1-4alkyl, CN, and halogen; g is 0, 1, 2, or 3; R7is selected from the group consisting of H, C1-4 alkyl, CN, and halogen, 64 171560744182219.00268 R8is H, or C1-4alkyl, wherein s is 0, 1, 2, 3 or 4.

2. The compound or the pharmaceutically acceptable salt thereof of claim 1, wherein B forms a urea or an amide linkage with L1.

3. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-2, wherein L1comprises 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, NH-3-6 membered cycloalkyl or NH-3-6 membered heterocycloalkyl, wherein the NH of NH-3-6 membered cycloalkyl or NH-3-6 membered heterocycloalkyl forms a linkage with B.

4. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-3, wherein L2comprises O or NH bonded to an optionally substituted ring selected from the group consisting of phenyl, 5-6 membered cycloalkyl, 5-6 membered heterocycloalkyl, 5-6 membered heteroaryl.

5. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-4, wherein L2comprises O or NH bonded to an optionally substituted ring selected from the group consisting of phenyl and 6 membered heteroaryl.

6. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-5, wherein L1-L2comprises NH-cyclohexyl-O-phenyl or cyclohexyl-NH-6-membered heteroaryl, wherein the phenyl and heteroaryl are optional substituted, wherein L1-L2is bonded to B via NH of NH- cyclohexyl -O-phenyl or cyclohexyl of cyclohexyl-NH-6- membered heteroaryl, wherein L2optionally further comprises one or more of C1-6 alkylene, C1-6 alkyleneC(O), and C1-6 alkyleneNH.

7. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-6, or, which is bonded to B via the atom with a star. 65 171560744182219.00268 8. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-3, wherein wherein L1comprises NH-3-6 membered cycloalkyl or NH-3-6 membered heterocycloalkyl, wherein the NH is bonded to B.

9. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-9, wherein C is C(O) and forms an amide linkage with L3.

10. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-9, wherein (L3)m comprises at least one of 3-6 membered cycloalkyl and 3-6 membered heterocycloalkyl.

11. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-10, wherein (L3)m is selected from the group consisting of66 171560744182219.00268 12. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-11, wherein A is phenyl substituted one or more substituents selected from the group consisting of O-haloC1-4 alkyl, haloC1-4 alkyl, C1-4 alkyl, CN, OC1-4 alkyl, O-haloC1-4 alkyl, CN, and halogen.

13. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-11, wherein A is benzyl substituted with one or more substituents selected from the group consisting of O-haloC1-4 alkyl, haloC1-4 alkyl, C1-4 alkyl, CN, OC1-4 alkyl, O-haloC1-4 alkyl, CN, halogen.

14. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-13, wherein D is selected from the group consisting ofbonded to E via the atom with a star, wherein D is optionally substituted with O-haloC1-4 alkyl, haloC1-4 alkyl, C1-4 alkyl, CN, OC1-4 alkyl, O-haloC1-4 alkyl, CN, halogen.

15. The compound or the pharmaceutically acceptable salt thereof of any one of claims 1-13, wherein D-E are selected from the group consisting of ,,,, , 67 171560744182219.00268, and, wherein each of the above structures is optionally substituted.

16. The compound or the pharmaceutically acceptable salt thereof of any one of the preceding claims, which is selected from the group consisting of68 171560744182219.00268 ,, 69 171560744182219.00268 O O H O O O N O N O O H OHN NO, O NH O N O O H H O O N O N O O N NO H H , O NH O O N O O H O O O N N O O N N H H H O O O O N H HN N N N O N O OO , O NH O O O NH N H O O O N OCF3N O N O H ONN NO OH H , O , O NH O H H O N N N H ON O O O ON O OCF3O , N O OCF O3O O HN N N N N N F O O H H O , O OCF O O N O3HN N N O N N N F O H H , O OCF O N O3HN N N N N F O N O H H O , O CF O N O3HN N N N N N O O H H O , 70 171560744182219.00268 O, 71 171560744182219.00268.

17. A pharmaceutical composition comprising a therapeutically effective amount of a compound or the pharmaceutically acceptable salt thereof of any one of claims 1-16.

18. A method of treating a disease associated with upregulated soluble epoxide hydrolase (sEH) in a subject, comprising administering to the subject in need thereof a therapeutically effective amount of the compound of any one of claims 1-16.

19. The method of claim 18, wherein the disease is selected from the group consisting of diabetes, neuropathic pain, chronic obstructive pulmonary disease (COPD), metabolic disorders, hyperlipidemia, Alzheimer's disease, Parkinson's disease, cardiovascular diseases, autism spectrum disorder (ASD) and schizophrenia, stroke, neuroinflammation, stroke, depression, alcohol associated liver disease, idiopathic pulmonary fibrosis (IPF), hypertension, asthma, diabetes, obesity, diabetic retinopathy, cardiac hypertrophy, pancreatic inflammation, pancreatic fibrosis, pulmonary fibrosis, renal fibrosis, chronic kidney disease, scarring, cisplatin toxicity, chemotherapy induced pain, atrial fibrillation, colitis, inflammatory bowel disease (IBD), sepsis (ARDS), metabolic dysfunction-associated steatohepatitis (MASH), osteoarthritis, rheumatoid arthritis, hardening of arteries, vascular inflammation, cardiac hypertrophy, atrial fibrosis, cardiac arrythmia, heat failure, myocardial infraction, traumatic brain injury, stroke, epilepsy, schizophrenia, depression, and inflammatory pain.

20. The method of claim 18, further comprising administering to the subject an agent selected from the group consisting of celecoxib, pioglitazone, bardoxolone, sildenafil, pirfenidone, infliximab, semaglutide, exenatide, liraglutide, dulaglutide, lixisenatide, efpeglenatide, fenofibrate, rosiglitazone, dexamethasone, methotrexate, simvastatin, minocycline, losartan, gabapentin. 72 171560744

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