An inhibitor of soluble epoxide hydrolase for use in the treatment of cancer cachexia

Abstract

Provided herein are methods of preventing, suppressing, or treating cancer cachexia in a subject as well as methods of prolonging survival of a subject in need of cancer cachexia treatment. Such methods include administering an inhibitor of soluble epoxide hydrolase (sEH).

Description

AN INHIBITOR OF SOLUBLE EPOXIDE HYDROLASE FOR USE IN THE TREATMENT OF CANCER CACHEXIACROSS-REFERENCES TO RELATED APPLICATIONS

[0001] NOT APPLICABLESTATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT

[0002] This invention was made with Government support under Grant No. ES030443 awarded by the National Institutes of Health (NIH). The Government has certain rights in the invention.BACKGROUND OF THE INVENTION

[0003] Cancer cachexia is a devastating syndrome characterized by progressive muscle wasting and hyperinflammation Cancer cachexia leads to organ failure, immunosuppression, and results in the death of about 20% of cancer patients (Al-Sawaf O et al. Body composition and lung cancer-associated cachexia in TRACERx. Nat Med. 2023 29(4): 846-858). Cancer cachexia has been reported to have a higher incidence rate in patients with gastric, pancreatic, lung, esophageal, colorectal, and head and neck cancer.

[0004] Despite the prevalence of cancer cachexia, the underlying mechanisms remain poorly characterized, and there are no effective treatments for cancer cachexia. Indeed, to date, there are no FDA-approved therapies to treat cancer cachexia.

[0005] As such, there is a need in the art to identify effective treatments for cancer cachexia. The present disclosure addresses these needs and provides related advantages as well.BRIEF SUMMARY

[0006] Provided herein are methods of preventing, suppressing, or treating cancer cachexia in a subject, the method comprising administering to the subject an inhibitor of soluble epoxide hydrolase (sEH).

[0007] Also provided herein are methods of prolonging survival of a subject in need of cancer cachexia treatment, the method comprising administering to the subject an inhibitor of soluble epoxide hydrolase (sEH).1INCORPORATED BY REFERENCE (RULE 20.6)

[0008] In some embodiments, the sEH inhibitor is a compound of Formula IIwherein the variable positions are as defined herein.

[0009] In some embodiments, the compound of Formula II has the structureor a pharmaceutically acceptable salt thereof.

[0010] Other objects, features, and advantages of the present disclosure will be apparent to one of skill in the art from the following detailed description and figures.BRIEF DESCRIPTION OF THE DRAWINGS

[0011] FIG. 1A-E shows (A) Relative mRNA expression, isolated from murine gastrocnemius muscles, of soluble epoxide hydrolase (ephx2) was measured by RT-qPCR and normalized by GAPDH. Non-tumor bearing (NTB) and pancreatic cancer (KPCY) groups were compared (n = 6 mice / group). (B) Protein was isolated from the gastrocnemius muscles of mice from the NTB and KPCY groups for western blot to determine the expression of soluble epoxide hydrolase (sEH). (C) sEH expression was quantified and normalized by GAPDH (n = 3 mice / group). (D) IHC using soluble epoxide hydrolase antibody (sEH, Santa Cruz Biotechnology, Catalog # sc-166961) was used to stain for the presence in paraffin-embedded munne gastrocnemius muscle. Representative images are at 20X objective. Scale bars are 50um. (E) Quantification of IHC was performed in ImageJ by calculating the percent area of positive staining per 20X fields. Ten fields were used for quantification per sample (n = 6 mice / group). The data are presented as the mean ± SEM. Unpaired Student’s t test was used. *p<0.05 and **p < 0.01.

[0012] FIG. 2A-D show (A) Percent survival was recorded between the vehicle-treated KPCY and the TPPU-treated mice for 40 days post-injection before sacrifice. All nontreated mice died at day 26 of study. TPPU was administered via a mini-osmotic pump at 5 mg / kg2INCORPORATED BY REFERENCE (RULE 20.6)while controls received PBS administration by mini-osmotic pump (n = 5 mice / group). (B) Percent survival was recorded between the vehicle-treated KPCY and the EC5026-treated mice for 400 days post-injection. EC5026 was administered in the drinking water at 5 mg / kg while controls received normal drinking water (n = 7 mice / group). (C) Percent survival was recorded between two groups of fatl+ / - mice: 1) KPCY-injected mice with normal drinking water and 2) KPCY-injected mice with EC5026 in their drinking water at 5 mg / kg (n = 5 mice / group). This survival study lasted until day 40 post-injection. (D) Percent survival was recorded between wildtype mice injected with KPCY and genetic whole-body ablation (KO) for sEH mice that were also injected with KPCY (n = 5 mice / group). All control mice died at day 25 of study while KO mice lasted until day 33 post-injected. A log-rank (Mantel-Cox) test and Gehan-Breslow-Wilcoxon test were utilized to test for significance. **p<0.01 and ***p < 0.001.

[0013] FIG. 3A-I shows (A) Body weight (grams) was measured at time of sacrifice (20 days post-injection). Gastrocnemius muscles, (B) right and (C) left, tibialis anterior muscles, (D) right and (E) left, (F) brain, (G) spleen, (H) liver, and (I) heart (grams) were normalized to body weight (n = 15 mice / group) and compared between groups: non-tumor bearing (NTB) vs. KPCY vehicle-treated controls vs. KPCY treated with EC5026. The data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.001, ns = no significance.

[0014] FIG. 4A-H shows that mice treated with EC5026 significantly reduced mRNA expression levels of (A) IL-6 and (B) NF-kB, (C) Cox-2, (D) Alox5, (E) Aloxl2, (F) Cyp2c65, and (G) LTB4R. KPCY vehicle-treated controls had increased expression levels in the mentioned markers comparatively. (H) Mice treated with EC5026 had increased mRNA expression levels of GPR18 compared to vehicle-treated controls. The mentioned RT-qPCR was performed on murine gastrocnemius tissues (n = 6 mice / group). The data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.001, ns = no significance.

[0015] FIG. 5A-H (A) Relative mRNA expression of soluble epoxide hydrolase (ephx2) measured by RT-qPCR was normalized by GAPDH (n = 6 mice / group). (B) Protein was isolated from the gastrocnemius muscles of mice for western blot to determine the expression3INCORPORATED BY REFERENCE (RULE 20.6)of soluble epoxide hydrolase and GAPDH (loading control). (C) sEH expression normalized by GAPDH (n = 3 mice / group). (D) Immunohistochemistry using soluble epoxide hydrolase antibody (sEH, Santa Cruz Biotechnology', Catalog # sc- 166961) was used to stain for presence in paraffin-embedded munne gastrocnemius muscle and representative pictures (20X) were taken for the gastrocnemius, tibialis anterior, spleen, and heart. Scale bar is 50um. Quantification of sEH IHC for the (E) gastrocnemius, (F) tibialis anterior, (G) spleen, and (H) heart tissue samples were performed using ImageJ by calculating the percent area of positive staining per 20X field (10 fields per sample) (n = 6 mice / group). The data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. **P < 0.01, ***P<0.001, ns = no significance.

[0016] FIG. 6A-I shows (A-D) The percentage of infiltrating F4 / 80+ macrophages (CD45+ CD1 lb+ F4 / 80+) increased in the brain, heart, muscle, and spleen tissue of KPCY vehicle-treated mice compared to NTB. KPCY mice treated with EC5026 showed a marked decrease in the percentage of infiltrating macrophages (CD45+ CD1 lb+ F4 / 80+) (n=10 mice / group). (E-F) The percentage of infiltrating NK cells (CD45+ CD3- NK1. 1+) increased in mice treated with EC5026 compared to vehicle-treated controls. (G-H) The percentage of infiltrating cytotoxic T cells (CD45+ CD3+ CD8+) increased in heart and spleen tissue of KPCY mice treated with EC5026 compared to NTB and KPCY vehicle-treated controls (n=10 mice / group). (I) Electronic gating strategy of the percentage of immune cell populations in murine tissue samples based on unique surface markers. The data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.001, ns = no significance.DETAILED DESCRIPTIONI. General

[0017] As descnbed above, a subset of cancer patients present with cancer induced cachexia. A significant number of these patients die due to cancer cachexia. As such, there is an urgent need to identify new therapeutic approaches that can prevent, suppress, or treat cancer cachexia.4INCORPORATED BY REFERENCE (RULE 20.6)

[0018] We hypothesized that pharmacological inhibition of the sEH may prevent cancer cachexia via the resolution of inflammation. The present disclosure demonstrates that administration of an inhibitor of soluble epoxide hydrolase (sEH) reduced organ wasting, improved body composition, and enhanced survival rates, indicating that administration of an sEH inhibitor can prevent, suppress, or treat cancer cachexia.IL Definitions

[0019] Units, prefixes, and symbols are denoted in their Systeme International de Unites (SI) accepted form. Numeric ranges are inclusive of the numbers defining the range. The headings provided herein are not limitations of the various aspects or embodiments, which can be had by reference to the specification as a whole. Accordingly, the terms defined immediately below are more fully defined by reference to the specification in its entirety. Terms not defined herein have their ordinary meaning as understood by a person of skill in the art.

[0020] The terms “about” and “approximately” shall generally mean an acceptable degree of error for the quantity measured given the nature or precision of the measurements.Typical, exemplary degrees of error are within 20 percent (%), preferably within 10%, and more preferably within 5% of a given value or range of values. Alternatively, and particularly in biological systems, the terms “about” and “approximately” may mean values that are within an order of magnitude, preferably within 5-fold and more preferably within 2- fold of a given value. Numerical quantities given herein are approximate unless stated otherwise, meaning that the term “about” or “approximately” can be inferred when not expressly stated.

[0021] The term "alkyl", by itself or as part of another substituent, means, unless otherwise stated, a straight or branched chain hydrocarbon group, having the number of carbon atoms designated (i.e. Ci-s means one to eight carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n- octyl, and the like. The term "alkenyl" refers to an unsaturated alkyl group having one or more double bonds. Similarly, the term "alkynyl" refers to an unsaturated alkyl group having one or more triple bonds. Examples of alkenyl groups include vinyl, 2-propenyl, crotyl, 2- isopentenyl, 2-(butadienyl), 2,4-pentadienyl and 3-(l,4-pentadienyl). Examples of alkynyl groups include ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs and isomers. The term "cycloalkyl" refers to hydrocarbon rings having the indicated number of ring atoms5INCORPORATED BY REFERENCE (RULE 20.6)(e.g., C3-6cycloalkyl) and being fully saturated or having no more than one double bond between ring vertices. "Cycloalkyl" is also meant to refer to bicyclic and polycyclic hydrocarbon rings such as, for example, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The bicyclic or polycyclic rings may be fused, bridged, spiro or a combination thereof. The term "heterocycloalkyl" or “heterocyclyl” refers to a cycloalkyl group having the indicated number of ring members, and which contain from one to three heteroatoms selected from N, 0, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quatemized. The heterocycloalkyl may be a monocyclic, a bicyclic or a polycylic ring system. The bicyclic or polycyclic rings may be fused, bridged, spiro or a combination thereof. It is understood that the recitation for C4-12 heterocyclyl, refers to a group having from 4 to 12 ring members where at least one of the ring members is a heteroatom. Non limiting examples of heterocycloalkyl groups include pyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, tetrazolone, hydantoin, dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S -oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrhydrothiophene, quinuclidine, and the like. A heterocycloalkyl group can be attached to the remainder of the molecule through a ring carbon or a heteroatom.

[0022] The terms "alkoxy," "alkylamino" and "alkylthio" (or thioalkoxy) are used in their conventional sense, and refer to those alkyl groups attached to the remainder of the molecule via an oxygen atom, an amino group, or a sulfur atom, respectively. Additionally, for dialkylamino groups, the alkyl portions can be the same or different and can also be combined to form a 3-7 membered ring with the nitrogen atom to which each is attached. Accordingly, a group represented as -NRaRbis meant to include piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl and the like.

[0023] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluonne, chlorine, bromine, or iodine atom. Additionally, terms such as "haloalkyl," are meant to include monohaloalkyl and polyhaloalkyl. For example, the term "C1-4 haloalkyl" is meant to include trifluoromethyl, 2,2,2-trifluoroethyl, 4- chlorobutyl, 3 -bromopropyl, and the like.

[0024] The term “hydroxyalkyl” or “alkyl-OH” refers to an alkyl group, as defined above, where at least one (and up to three) of the hydrogen atoms is replaced with a hydroxy group. As for the alkyl group, hydroxyalkyl groups can have any suitable number of carbon atoms,6INCORPORATED BY REFERENCE (RULE 20.6)such as Ci-6. Exemplary' hydroxy alkyl groups include, but are not limited to, hydroxymethyl, hydroxy ethyl (where the hydroxy is in the 1- or 2-position), hydroxypropyl (where the hydroxy is in the 1-, 2- or 3-position), and 2,3-dihydroxypropyl.

[0025] The term "aryl" means, unless otherwise stated, an aromatic hydrocarbon group which can be a single ring or multiple rings (up to three rings) which are fused together or linked covalently. Non-limiting examples of aryl groups include phenyl, naphthyl and biphenyl. The term "heteroaryl" refers to aryl groups (or rings) that contain from one to five heteroatoms selected from N, 0, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quatemized. A heteroaryl group can be attached to the remainder of the molecule through a heteroatom. Non-limiting examples of heteroaryl groups include pyridyl, pyridazinyl, pyrazinyl, pyrimindinyl, triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalaziniyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl, isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridinyl, thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridines, benzothiaxolyl, benzofuranyl, benzothienyl, indolyl, quinolyl, isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furyl, thienyl and the like.

[0026] As used herein, the term "heteroatom" is meant to include oxygen (0), nitrogen (N), sulfur (S), phosphorus (P) and silicon (Si).

[0027] The disclosure herein further relates to prodrugs and bioisosteres thereof. Suitable bioisosteres, for example, will include carboxylate replacements (phosphonic acids, phosphinic acids, sulfonic acids, sulfmic acids, and acidic heterocyclic groups such as tetrazoles). Suitable prodrugs will include those conventional groups known to hydrolyze and / or oxidize under physiological conditions to provide a compound of Formula I.

[0028] The terms "patient" and “subject” include primates (especially humans), domesticated companion animals (such as dogs, cats, horses, and the like) and livestock (such as cattle, pigs, sheep, and the like).

[0029] As used herein, the term "treating" or "treatment" encompasses both diseasemodifying treatment and symptomatic treatment, either of which may be prophylactic (z.e., before the onset of symptoms, in order to prevent, delay or reduce the severity of symptoms) or therapeutic (i.e., after the onset of symptoms, in order to reduce the severity and / or duration of symptoms).7INCORPORATED BY REFERENCE (RULE 20.6)

[0030] " cis-Epoxyeicosatrienoic acids" ("EETs") are biomediators synthesized by cytochrome P450 epoxygenases.

[0031] "Epoxide hydrolases" ("EH;" EC 3.3.2.3) are enzymes in the alpha beta hydrolase fold family that add water to 3-membered cyclic ethers termed epoxides.

[0032] " Soluble epoxide hydrolase" ("sEH") is an epoxide hydrolase which in endothelial and smooth muscle cells converts EETs to dihydroxy derivatives called dihydroxy eicosatrienoic acids ("DHETs"). The cloning and sequence of the murine sEH is set forth in Grant et al., J. Biol. Chem. 268(23):17628-17633 (1993). The cloning, sequence, and accession numbers of the human sEH sequence are set forth in Beetham et al., Arch. Biochem. Biophys. 305(1): 197-201 (1993). The evolution and nomenclature of the gene is discussed in Beetham et al., DNA Cell Biol. 14(1):61 -71 (1995). Soluble epoxide hydrolase represents a single highly conserved gene product with over 90% homology between rodent and human (Arand et al., FEBS Lett., 338:251-256 (1994)). Unless otherwise specified, as used herein, the terms "soluble epoxide hydrolase" and "sEH" refer to human sEH.

[0033] Unless otherwise specified, as used herein, the term "sEH inhibitor" (also abbreviated as "sEHI") refers to an inhibitor of human sEH. Preferably, the inhibitor does not also inhibit the activity of microsomal epoxide hy drolase by more than 25% at concentrations at which the inhibitor inhibits sEH by at least 50%, and more preferably does not inhibit sEH by more than 10% at that concentration. For convenience of reference, unless otherwise required by context, the term "sEH inhibitor" as used herein encompasses prodrugs which are metabolized to active inhibitors of sEH. Further for convenience of reference, and except as otherwise required by context, reference herein to a compound as an inhibitor of sEH includes reference to derivatives of that compound (such as an ester of that compound) that retain activity as an sEH inhibitor.

[0034] Cytochrome P450 ("CYP450") metabolism produces cA-epoxydocosapentaenoic acids (“EpDPEs”) and cA-epoxy eicosatetraenoic acids (“EpETEs”) from docosahexaenoic acid ("DHA") and eicosapentaenoic acid ("EP A"), respectively. These epoxides are known endothelium-derived hyperpolarizing factors ("EDHFs"). These EDHFs, and others yet unidentified, are mediators released from vascular endothelial cells in response to acetylcholine and bradykinin, and are distinct from the NOS- (nitric oxide) and COX-derived (prostacyclin) vasodilators. Overall cytochrome P450 (CYP450) metabolism of8INCORPORATED BY REFERENCE (RULE 20.6)polyunsaturated fatty acids produces epoxides, such as EETs. 14(15)-EpETE, for example, is derived via epoxidation of the 14,15-double bond of EPA and is the co-3 homolog of 14(15)- EpETrE (“14(15)EET”) derived via epoxidation of the 14,15-double bond of arachidonic acid.

[0035] The term “therapeutically effective amount” refers to that amount of the compound being administered sufficient to prevent, mitigate, decrease, reverse the development of one or more of the symptoms of the disease, condition or disorder being treated.

[0036] The terms “sustained release” and “extended release” are used in their conventional sense to refer to a drug formulation that provides for gradual release of a drug over an extended period of time, for example, 12 hours or more, and that preferably, although not necessarily, results in substantially steady-state blood levels of a drug over an extended time period.

[0037] The terms “systemic administration” and “systemically administered” refer to a method of administering agent to a mammal so that the agent / cells is delivered to sites in the body, including the targeted site of pharmaceutical action, via the circulatory system.Systemic administration includes, but is not limited to, oral, intranasal, rectal and parenteral ( / .e., other than through the alimentary tract, such as intramuscular, intravenous, intra-arterial, transdermal and subcutaneous) administration.

[0038] The phrase "cause to be administered" refers to the actions taken by a medical professional (e.g. , a physician), or a person controlling medical care of a subject, that control and / or permit the administration of the agent(s) / compound(s) / cell(s) at issue to the subject. Causing to be administered can involve diagnosis and / or determination of an appropriate therapeutic or prophylactic regimen, and / or prescribing particular agent(s) / compounds / cell(s) for a subject. Such prescribing can include, for example, drafting a prescription form, annotating a medical record, and the like.

[0039] The terms “patient,” “subject” or “individual” interchangeably refers to a non- human mammal, including primates (e.g., macaque, pan troglodyte, pongo), a domesticated mammal (e.g., felines, canines), an agricultural mammal (e.g., bovine, ovine, porcine, equine) and a laboratory mammal or rodent (e.g., rattus, murine, lagomorpha, hamster).9INCORPORATED BY REFERENCE (RULE 20.6)III. Methods of preventing, suppressing, or treating cancer cachexia in a subject

[0040] Provided herein are methods of preventing, suppressing, or treating cancer cachexia in a subject, the method comprising administering to the subject an inhibitor of soluble epoxide hydrolase (sEH).

[0041] In some aspects, provided herein are methods prolonging survival of a subject in need of cancer cachexia treatment, the method comprising administering to the subject an inhibitor of soluble epoxide hydrolase (sEH).

[0042] Subjects who may benefit from these treatment methods are those who have been diagnosed with cancer cachexia or are suspected of having cancer cachexia.

[0043] In some embodiments, the cancer is bladder, ovarian, cervical, breast, testicular, prostate, head and neck, oral, esophageal, gastric, lung, pancreatic, skin, leukemia, colon or colorectal cancer. In some embodiments, the subject has bladder cancer.

[0044] In some embodiments, the cancer is melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma, Hodgkin’s lymphoma, head and neck squamous cell carcinoma (HNSCC), merkel cell carcinoma, a microsatellite instability-high (MSI-H) cancer, colorectal cancer, hepatocellular carcinoma, primary mediastinal large B-cell lymphoma (PMBCL), small cell lung cancer (SCLC), cutaneous squamous-cell carcinoma, basal cell carcinoma, mismatch repair deficient (dMMR) cancer, Endometrial Carcinoma, Esophagus cancer, Malignant Pleural Mesothelioma, high tumor mutation burden (TMB-H) cancers. In some embodiments, the cancer is melanoma.

[0045] In some embodiments, the subject has a hematologic malignancy, which include without limitation lymphomas (such as but not limited to, non-Hodgkin’s lymphoma, including Burkitt’s lymphoma, and Hodgkin’s lymphoma, as well as all subtypes associated with each), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), and adult T-cell leukemia lymphoma.

[0046] In some embodiments, the subject has a hematologic malignancy such as multiple myeloma or plasmocytoma.10INCORPORATED BY REFERENCE (RULE 20.6)

[0047] In some embodiments, the subject has a sarcoma. In some embodiments the sarcoma includes without limitation rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma and Ewing’s sarcoma.

[0048] In some embodiments, the subject has a urogenital cancers, which include without limitation labial carcinoma, cervix carcinoma, uterine corpus carcinoma, endometrium carcinoma, chorion carcinoma, , testis carcinoma, seminoma, urinary carcinoma, kidney carcinoma, renal carcinoma, and adenocarcinoma (including adenocarcinomas of the vagina, cervix, and urachus).

[0049] In some embodiments, the subject has a nervous and sensory system cancer, which include without limitation neuroblastoma, brain tumors, meningioma, ependymoma, medulloblastoma, peripheral neuroectodermal tumors, glioblastoma, astrocytoma, oligodendroglioma and retinoblastoma.

[0050] In some embodiments, the subject has an endocrine and glandular tissue cancer, which include without limitation pancreatic carcinoma, medullary thyroid carcinoma, follicular thy roid carcinoma, anaplastic thyroid carcinoma, papillary thyroid carcinoma, pheochromocytoma, adrenal tumors and adenocarcinoma.

[0051] In some embodiments, the subject has a hepatic cancer, which include without limitation hepatocellular carcinoma.

[0052] In some embodiments, the subject has a skin cancer, which include without limitation melanoma, basal cell carcinoma, squamous cell carcinoma and choroids melanoma.IV. Inhibitors of soluble epoxide hydrolase (sEH inhibitors)

[0053] Inhibitors of sEH have been previously disclosed. Each of these compounds are intended to be within the scope of the current disclosure.

[0054] In some embodiments, the sEH inhibitor is a compound of Formula IIwhereinINCORPORATED BY REFERENCE (RULE 20.6)R4is -OCF3 or -CF3; each R4ais H, halogen, Ci-6 alkyl, CM alkoxy, Ci-6 haloalkyl, C1-6 haloalkoxy, -O-Ce- 12 aryl, heterocycloalkyl, -OH, -NO2, or -C(O)OR4b, wherein each heterocycloalkyl has 5 to 6 ring members and 1 to 3 heteroatoms each independently N, 0, or S;R5is CM alkyl, CM haloalkyl, Ci-6 hydroxyalkyl, C alkoxy, C haloalkoxy, -X5- C3-6 cycloalkyl, -X5-heterocycloalkyl, and -X5-heteroaryl, wherein each heterocycloalkyl has 3 to 6 ring members and 1 to 3 heteroatoms each independently N, 0, or S, and each heteroaryl has 5 to 6 ring members and 1 to 3 heteroatoms each independently N, 0 or S;R5is substituted with 0, 1, 2, or 3 substituents, each independently CM alkyl, C haloalkyl, -OH, -C(O)OR5a, or -Ci- -alkylene-C(O)OR5a;X5is a bond or C1-3 alkylene;R4band R5aare each independently H or Ci-6 alkyl; and subscript m is 0, 1 or 2.

[0055] In some embodiments, the compound of Formula II has the structure of Formula Ila

[0056] In some embodiments, R4in Formula II or Ila is -OCF3.

[0057] In some embodiments, R4in Formula II or Ila is -CF3.

[0058] In some embodiments, m is 1 and R4ain Formula II or Ila is -CF3, Cl, Br, F, or -OCF3.

[0059] In some embodiments, m is 1 and R4ain Formula II or Ila is F.

[0060] In some embodiments, R5in Formula II or Ila is Ci-6 alkyl, C haloalkyl, C hydroxyalkyl, CM alkoxy, CM haloalkoxy, C3-6 cycloalkyl, heterocycloalkyl, or heteroaryl, wherein each heterocycloalkyl has 3 to 6 ring members and 1 to 3 heteroatoms each independently N, O, or S, and each heteroaryl has 5 to 6 ring members and 1 to 3 heteroatoms each independently N, O or SINCORPORATED BY REFERENCE (RULE 20.6)

[0061] In some embodiments, R5in Formula II or Ila is Ci-6 alkyl, Ci-6 haloalkyl or Ci-6 hydroxy alkyl.

[0062] In some embodiments, the compounds of Formula II is a compound listed in Table 2.Table 1: Compounds of Formula II13INCORPORATED BY REFERENCE (RULE 20.6)INCORPORATED BY REFERENCE (RULE 20.6)

[0063] In some embodiments, the compound of Formula II has the structureor a pharmaceutically acceptable salt thereof.

[0064] In some embodiments, the compound of Formula II has the structure

[0065] In some embodiments, the compound of Formula II has the structureor a pharmaceutically acceptable salt thereof.INCORPORATED BY REFERENCE (RULE 20.6)

[0066] In some embodiments, the compound of Formula II has the structure

[0067] The compounds of Formula II may exist as salts. The present invention includes such salts. Typically, the salts used are pharmaceutically acceptable salts.

[0068] Pharmaceutically acceptable salts include salts of the active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compounds of Formula II contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of Formula II contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydnodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge etal., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of Formula II contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0069] The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.INCORPORATED BY REFERENCE (RULE 20.6)

[0070] Certain compounds of Formula II can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present invention. Certain compounds of Formula II may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[0071] Certain compounds of Formula II possess asymmetric carbon atoms (optical centers) or double bonds; the enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisometric forms that may be defined, in terms of absolute stereochemistry, as(R)-or (S)- or, as (D)- or (L)- for amino acids, and individual isomers are encompassed within the scope of the present invention. The compounds of Formula II do not include those which are known in art to be too unstable to synthesize and / or isolate. The present invention is meant to include compounds in racemic and optically pure forms. Optically active (R)- and(S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.

[0072] Isomers include compounds having the same number and kind of atoms, and hence the same molecular weight, but differing in respect to the structural arrangement or configuration of the atoms.

[0073] It will be apparent to one skilled in the art that certain compounds of this invention may exist in tautomeric forms, all such tautomeric forms of the compounds being within the scope of the invention. Tautomer includes one of two or more structural isomers which exist in equilibrium and which are readily converted from one isomeric form to another.

[0074] Unless otherwise stated, structures depicted herein are also meant to include all stereochemical forms of the structure; i.e., the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the invention.

[0075] Unless otherwise stated, the compounds of Formula II may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds of Formula II may be radiolabeled with radioactive isotopes, such as for example deuterium (2H), tritium (3H), iodine-125 (125I), carbon-13 (13C), or carbon-14 (14C). All isotopic variations of the compounds of Formula II, whether radioactive or not, are encompassed within the scope of the present invention.17INCORPORATED BY REFERENCE (RULE 20.6)

[0076] In addition to salt forms, the compounds of Formula II can be prepared as prodrugs. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of Formula II. Additionally, prodrugs can be converted to the compounds of Formula II by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of Formula II when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0077] The compounds of Formula II can be made by a variety of methods known in the art.

[0078] In some embodiments the sEH inhibitor is GSK2256294AV. Formulation and Administration

[0079] The agents for administration in the present methods (e.g., an sEH inhibitor) can be formulated and administered via known routes in the art. The pharmacologically active agent can be prepared and administered in a wide variety of oral, parenteral and topical dosage forms. In some embodiments, the active agent is formulated in a pharmaceutically acceptable carrier or excipient.

[0080] For preparing pharmaceutical compositions, the pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances which may also act as diluents, flavoring agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material.

[0081] In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active component. In tablets, the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain from 5% or 10% to 70% of the active compound. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium18INCORPORATED BY REFERENCE (RULE 20.6)carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term “preparation” is intended to include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component with or without other carriers, is surrounded by a carrier, which is thus in association with it.Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0082] For preparing suppositories, a low melting wax, such as a mixture of fatty acid glycerides or cocoa butter, is first melted and the active component is dispersed homogeneously therein, as by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool, and thereby to solidify.

[0083] Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water / propylene glycol solutions. For parenteral injection, liquid preparations can be formulated in solution in aqueous polyethylene glycol solution. Transdermal administration can be performed using suitable carriers. If desired, apparatuses designed to facilitate transdermal delivery can be employed. Suitable carriers and apparatuses are well known in the art, as exemplified by U.S. Patent Nos. 6,635,274, 6,623,457, 6,562,004, and 6,274,166.

[0084] Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizers, and thickening agents as desired. Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active components in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents.

[0085] Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration. Such liquid forms include solutions, suspensions, and emulsions. These preparations may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.

[0086] A variety of solid, semisolid and liquid vehicles have been known in the art for years for topical application of agents to the skin. Such vehicles include creams, lotions, gels, balms, oils, ointments and sprays. See, e.g., Provost C. "Transparent oil-water gels: a review," Int J Cosmet Sci. 8:233-247 (1986), Katz and Poulsen, Concepts in biochemical19INCORPORATED BY REFERENCE (RULE 20.6)pharmacology, part I. In: Brodie BB, Gilette JR, eds. Handbook of Experimental Pharmacology. Vol. 28. New York, NY: Springer; 107-174 (1971), and Hadgcraft, "Recent progress in the formulation of vehicles for topical applications," Br J Dermatol., 81 : 386-389 (1972). A number of topical formulations of analgesics, including capsaicin (e.g., Capsin®), so-called "counter-irritants" (e.g., Icy-Hot®, substances such as menthol, oil of wintergreen, camphor, or eucalyptus oil compounds which, when applied to skin over an area presumably alter or off-set pain in joints or muscles served by the same nerves) and salicylates (e.g. BenGay®), are known and can be readily adapted for topical administration. It is presumed that the person of skill is familiar with these various vehicles and preparations and they need not be described in detail herein.

[0087] The pharmacologically active agents can be mixed into such modalities (creams, lotions, gels, etc.) for topical administration. In general, the concentration of the agents provides a gradient which drives the agent into the skin. Standard ways of determining flux of drugs into the skin, as well as for modifying agents to speed or slow their deliver}' into the skin are well known in the art and taught, for example, in Osborne and Amann, eds., Topical Drug Delivery Formulations, Marcel Dekker, 1989. The use of dermal drug delivery agents in particular is taught in, for example, Ghosh et al., eds., Transdermal and Topical Drug Delivery Systems, CRC Press, (Boca Raton, FL, 1997).

[0088] In some embodiments, the active agent is formulated in a cream. Typically, the cream comprises one or more hydrophobic lipids, with other agents to improve the "feel" of the cream or to provide other useful characteristics.

[0089] In other embodiments, the active agent is formulated in a lotion. Typical lotions comprise, for example, water, mineral oil, petrolatum, sorbitol solution, stearic acid, lanolin, lanolin alcohol, cetyl alcohol, glyceryl stearate / PEG-100 stearate, triethanolamine, dimethicone, propylene glycol, microcrystalline wax, tri (PPG-3 myristyl ether) citrate, disodium EDTA, methylparaben, ethylparaben, propylparaben, xanthan gum, butylparaben, and methyldibromo glutaronitrile.

[0090] In some embodiments, the agent is in an oil, such as jojoba oil. In some embodiments, the agent is, or agents are, in an ointment, which may, for example, white petrolatum, hydrophilic petrolatum, anhydrous lanolin, hydrous lanolin, or polyethylene glycol. In some embodiments, the agent is in a spray, which typically comprise an alcohol20INCORPORATED BY REFERENCE (RULE 20.6)and a propellant. If absorption through the skin needs to be enhanced, the spray may optionally contain, for example, isopropyl myristate.

[0091] Whatever the form in which the agent is administered (that is, whether by lotion, gel, spray, etc.), they are preferably administered at a dosage of about 0.01 mg to 10 mg per 10 cm2.

[0092] The pharmaceutical preparation is preferably in unit dosage form. In such form the preparation 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.

[0093] 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.

[0094] The dosage of a specific agent 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 of an sEH inhibitor is from about 0. 1 mg / kg to about 10 mg / kg, 25 mg / kg, 50mg / kg, 100 mg / kg, 150 mg / kg, 200 mg / kg, 250 mg / kg, 300 mg / kg, 350 mg / kg, 400 mg / kg, or 500 mg / kg body weight of the mammal.

[0095] Determination of an effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein. Generally, an efficacious or effective amount is determined by first administering a low dose or small amount of an agent and then incrementally increasing the administered dose or dosages 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, 13th Edition, 2017, supra,' in a Physicians’ Desk Reference (PDR), 71st Edition, 2017 (PDRNetwork); in Remington: The Science and Practice of Pharmacy, 21stEd., 2005, supra,' and in Martindale: The Complete Drug Reference, Sweetman, 2005, London: Pharmaceutical Press., and in21INCORPORATED BY REFERENCE (RULE 20.6)Martindale, Martindale: The Extra Pharmacopoeia, 31st Edition., 1996, Amer Pharmaceutical Assn, each of which are hereby incorporated herein by reference.

[0096] In some embodiments, the sEH inhibitor is administered orally.VI. Examples

[0097] The following examples are provided to illustrate but not limit the claimed invention.Example 1: Mouse Models of Cancer CachexiaMethods:

[0098] We investigated murine cancer cachexia models using genetically engineered pancreatic (KPC) and prostate (TRAMP Cl) tumor cell lines, and genetically engineered mice (transgenic adenocarcinoma of the mouse prostate-TRAMP). Cachectic mice were treated with human sEH inhibitor EC5026.Results:

[0099] An increase in sEH expression was observed in the spleen, liver, heart, gastrocnemius (GA), and tibialis anterior (TA) muscles compared to healthy controls by IHC and qPCR (n = 5 / group). KPC cells injected intraperitoneally induced a reduction in the weights of the spleen, liver, heart, brain, GA, and TA compared to healthy controls (n = 15 / group). KPC mice treated with EC5026 showed increased and comparable organ weights to healthy controls. In addition, treatment with EC5026 significantly improved survival rates in both KPC and TRAMP models. A sustained survival of over 250 days was observed postinjection in the KPC model (n=15 mice / group). In the TRAMP model, 5 / 5 of mice treated with EC5026 survived 230 days post-treatment compared to no survival of vehicle-treated mice (n=5). Mice treated with EC5026 showed a reduction in sEH expression in all tissues. Additionally, qPCR analysis of gastrocnemius tissue indicated that sEH inhibition attenuated pro-inflammatoiy markers and eicosanoid enzymes, evidenced by reduced expression of IL- 6, NF-kB, LTB4R, Cox-2, Alox5, Aloxl2, Cyp2j5, and Cyp2c65 comparatively (n=5) and increased pro-resolving receptors (e.g. RvD2 / GPR18). Immunoprofiling by flow cytometry (n=l 5 / group) revealed a significant increase in macrophages, CD8+ T cells, and NK cells, and a decrease in CD4+ T cells in EC5026-treated KPC mice across various cachexia tissues. KPC mice treated with EC5026 expressed similar percentages of immune cell populations compared to healthy controls in each tissue type.22INCORPORATED BY REFERENCE (RULE 20.6)Conclusions:

[0100] Thus, sEH inhibition may be a novel host-directed therapeutic approach to cancer cachexia by targeting the host immune response via stimulation of resolution of inflammation without toxicity or immunosuppression.

[0101] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, one of skill in the art will appreciate that certain changes and modifications may be practiced within the scope of the appended claims. In addition, each reference provided herein is incorporated by reference in its entirety to the same extent as if each reference was individually incorporated by reference. Where a conflict exists between the instant application and a reference provided herein, the instant application shall dominate.INCORPORATED BY REFERENCE (RULE 20.6)

Claims

WHAT IS CLAIMED IS:

1. A method of preventing, suppressing, or treating cancer cachexia in a subject, the method comprising administering to the subject an inhibitor of soluble epoxide hydrolase (sEH).

2. A method of prolonging survival of a subject in need of cancer cachexia treatment, the method comprising administering to the subject an inhibitor of soluble epoxide hydrolase (sEH).

3. The method of claim 1 or 2, wherein the sEH inhibitor isor a pharmaceutically acceptable salt thereof.

4. The method of any one of claims 1 to 3, wherein the subject has a cancer selected from the group consisting of: bladder, ovarian, cervical, breast, testicular, prostate, head and neck, oral, esophageal, gastric, lung, pancreatic, skin, leukemia, colon and colorectal cancer.

5. The method of any one of claims 4, wherein the subject has pancreatic cancer.

6. The method of any one of claims 4, wherein the subject has prostate cancer.

7. The method of any one of claims 4, wherein the subject has gastric cancer.

8. The method of any one of claims 4, wherein the subject has esophageal cancer.

9. The method of any one of claims 4, wherein the subject has colorectal cancer.

10. The method of any one of claims 4, wherein the subject has head and neck cancer.24INCORPORATED BY REFERENCE (RULE 20.6)11. The method of any one of claims 4, wherein the subject has lung cancer.

12. The method of any one of claims 1 to 11, wherein the subject is a mammal.

13. The method of claim 12, wherein the mammal is a human.25INCORPORATED BY REFERENCE (RULE 20.6)

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