WO2022076881A1

WO2022076881A1 - Methods of treating alzheimer's disease using slow tight binding soluble epoxide hydrolase inhibitors

Info

Publication number: WO2022076881A1
Application number: PCT/US2021/054264
Authority: WO
Inventors: Bruce D. Hammock; Sung Hee Hwang; Christophe Morisseau; Karen M. Wagner
Original assignee: The Regents Of The University Of California; Eicosis, Llc
Priority date: 2020-10-09
Filing date: 2021-10-08
Publication date: 2022-04-14

Abstract

Provided herein are methods of preventing, reducing, ameliorating, mitigating, slowing the progression and/or treating Alzheimer's disease using a tight binding soluble epoxide hydrolase (sEH) inhibitor, having slow dissociation rates and a long half-life of dissociation. In some embodiments, tight binding soluble epoxide hydrolase (sEH) inhibitors have an in vitro half-life of dissociation from human sEH protein of at least 5 minutes as determined by a Frster resonance energy transfer (FRET)-based competitive displacement assay. In some embodiments, the tight binding soluble epoxide hydrolase (sEH) inhibitors include are compounds of Formula I (I).

Description

METHODS OF TREATING ALZHEIMER'S DISEASE USING SLOW TIGHT

BINDING SOLUBLE EPOXIDE HYDROLASE INHIBITORS

CROSS-REFERENCES TO RELATED APPLICATIONS

[0001] This is an application claiming priority benefit under 35 U.S. C. § 119(e) of U.S. Provisional Application No. 63/089,795 filed October 9, 2020, which is herein incorporated by reference in its entirety for all purposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT

[0002] This invention was made with Government support under Grant Nos. 1R35ES030443- 01 and P42ES04699, awarded by the National Institutes of Health. This invention was made using the following grants: NIEHS SBIR Program R44ES025598, the NIH NINDS Blueprint Neurotherapeutics Network UH2NS094258, National Institute of Neurological Disorders and Stroke (NINDS) U54 NS079202-01 and the National Institutes of Drug Abuse (NIDA) UG3 DA048767, each of these being awarded to EicOsis. The Government has certain rights in the invention.

REFERENCE TO A "SEQUENCE LISTING," A TABLE, OR A COMPUTER PROGRAM LISTING APPENDIX SUBMITTED ON A COMPACT DISK

[0003] NOT APPLICABLE

BACKGROUND OF THE INVENTION

[0004] Alzheimer’s disease (AD) is a progressive illness characterized by memory deficit together with dysphasia (language disorder in which there is an impairment of speech and of comprehension of speech), dyspraxia (inability to coordinate and perform certain purposeful movements and gestures in the absence of motor or sensory impairments) and agnosia (inability to recognize objects, persons, sounds, shapes, or smells) attributable, in part, to involvement of the accumulation of proteolytic products of the amyloid precursor protein (APP), amyloid-p peptides (A|3), which form extracellular aggregates termed A|3 plaques and intraneuronal neurofibrillary tangles composed of hyperphosphorylated tau protein. [0005] Incidence of Alzheimer disease increases dramatically with the age. It is clinically characterized by a global decline of cognitive function that progresses slowly and leaves endstage patients bound to bed, incontinent and dependent on custodial care. Death occurs, on average, 9 years after diagnosis,

[0006] As mentioned above, the pathological hallmark of AD includes amyloid plaques containing A|3. For the last decade, two major hypotheses on the cause of AD have been proposed: the "amyloid cascade hypothesis", which states that the neurodegenerative process is a series of events triggered by the abnormal processing of the Amyloid Precursor Protein (APP), and the "neuronal cytoskeletal degeneration hypothesis", which proposes that cytoskeletal changes are the triggering events. The most widely accepted theory explaining AD progression remains the amyloid cascade hypothesis and AD researchers have mainly focused on determining the mechanisms underlying the toxicity associated with A|3. However, recent drug trials based on this hypothesis have not been successful.

[0007] Based on the understood pathologic mechanisms and brain involvement in AD, clinical interventions that might modify disease progression or address the underlying cause of the disease are assumed to be compositions that can readily penetrate the blood-brain barrier (BBB). The BBB is composed principally of cerebrovascular endothelial cells that separates circulating blood from the brain parenchyma, restricting the transport of substances such as drugs into the brain. There are also energy dependent pumps known as drug transporters which move some foreign compounds out of the brain.

[0008] There is no known cure of AD. Despite significant research efforts, only yielded a handful of approved clinical interventions have been approved by the FDA. These drugs generally treat the symptoms associated with AD, but they do not address the underlying cause of the disease. Even with sustained treatment, AD continues to progress in these patients. Overtime, the approved drugs become less effective.

[0009] As such, there is a need in the art to identify alternative medications that can prevent, reduce, ameliorate, mitigate, or treat AD. The present disclosure addresses these needs and provides related advantages as well. BRIEF SUMMARY OF THE INVENTION

[0010] In some aspects, provided herein are methods of preventing, reducing, ameliorating, mitigating, slowing the progression and/or treating Alzheimer’s disease in a human subject comprising administering to the human subject a therapeutically effective amount of a tight binding soluble epoxide hydrolase (sEH) inhibitor.

[0011] In some embodiments, the tight binding soluble epoxide hydrolase (sEH) inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 5 minutes as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay.

[0012] In some embodiments, the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 4 nM or lower as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay.

[0013] In some embodiments, a total daily dose of about 2 to 20 mg of the tight binding sEH inhibitor is administered to the human subject.

[0014] In some embodiments, the tight binding soluble epoxide hydrolase (sEH) inhibitor is a compound of Formula I

wherein, R¹, R², n, X¹, Y, and Z are as defined herein.

BRIEF DESCRIPTION OF THE DRAWINGS

[0015] NOT APPLICABLE

DETAILED DESCRIPTION OF THE INVENTION

I. General

[0016] The inventors of the present disclosure have discovered that Alzheimer’s disease can be effectively prevented, reduced, ameliorated, mitigated, and/or treated using a soluble epoxide hydrolase (sEH) inhibitor that has a slow dissociation rate from human sEH protein. The slow dissociation rate affords a long residence time of the inhibitor on the targeted soluble epoxide hydrolase (sEH) protein. Without being bound to any particular theory, it is believed that the slow dissociation rate provides a surprising bioaccumulation of these inhibitors in the brain (above what a person of skill in the art would anticipate based on the physical properties of sEH inhibitors). Thus, compounds with expected moderate or low BBB permeability unexpectedly can be used to treat Alzheimer’s disease. It is believed that the compounds described herein advantageously and unexpectedly avoid outward transport from the drug transporters within the blood-brain barrier. That is, the slow dissociation rate allows the compounds to stay bound to the targeted protein for long periods of time, staying in the subject system for significantly longer than expected based on separate metabolism studies.

[0017] Without being bound to any particular theory, it is further believed that soluble epoxide hydrolase (sEH) inhibitors that have a slow dissociation rate from human sEH protein will accumulate in tissues with high sEH protein, like the liver. The local accumulation of inhibitor provides a local increase in fatty acid epoxides that surprisingly also shifts the systemic (whole body) levels of fatty acid epoxides. The unexpected increased systemic levels of fatty acid epoxides can also surprisingly raise the titer of fatty acid epoxides that are transported to the brain, providing a positive benefit in the prevention, reduction, amelioration, mitigation, and/or treatment of Alzheimer’s disease.

II. Definitions

[0018] Units, prefixes, and symbols are denoted in their Systeme International d’Unites (SI) accepted form. Numeric ranges are inclusive of the numbers defining the range. Unless otherwise indicated, nucleic acids are written left to right in 5' to 3' orientation; amino acid sequences are written left to right in amino to carboxy orientation. 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.

[0019] "Soluble epoxide hydrolase" ("sEH"; EC 3.3.2.10) is an epoxide hydrolase which in cells converts EETs and other epoxyfatty acids (EpFA) to dihydroxy derivatives called dihydroxyeicosatri enoic 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.

[0020] The term “therapeutically effective amount” refers to that amount of the compound being administered sufficient to prevent or decrease the development of one or more of the symptoms of the disease, condition or disorder being treated.

[0021] “Alkyl” refers to a straight or branched, saturated, aliphatic radical having the number of carbon atoms indicated (i.e., Ci-Ce means one to six carbons). Alkyl can include any number of carbons, such as C1-C2, C1-C3, C1-C4, C1-C5, Ci-C₆, C1-C7, Ci-C₈, C1-C9, C1-C10, C2-C3, C₂- C₄, C2-C5, C₂-C₆, C3-C4, C3-C5, C3-C6, C4-C5, C₄-C₆ and C₅-C₆. For example, Ci-C₆ alkyl includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, hexyl, etc. Alkyl can also refer to alkyl groups having up to 20 carbons atoms, such as, but not limited to heptyl, octyl, nonyl, decyl, etc.

[0022] “Alkylene” refers to a straight or branched, saturated, aliphatic radical having the number of carbon atoms indicated (i.e., Ci-Ce means one to six carbons), and linking at least two other groups, i.e., a divalent hydrocarbon radical. The two moieties linked to the alkylene can be linked to the same atom or different atoms of the alkylene group. For instance, a straight chain alkylene can be the bivalent radical of -(CH2)_n-, where n is 1 , 2, 3, 4, 5 or 6. Representative alkylene groups include, but are not limited to, methylene, ethylene, propylene, isopropylene, butylene, isobutylene, sec-butylene, pentylene and hexylene.

[0023] “Cycloalkyl” refers to a saturated or partially unsaturated, monocyclic, fused bicyclic or bridged polycyclic ring assembly containing from 3 to 12 ring atoms, or the number of atoms indicated. Cycloalkyl can include any number of carbons, such as C3-C6, C4-C6, C5-C6, C3-C8, C4-C8, Cs-Cs, Ce-Cs, C3-C9, C3-C10, C3-C11, and C3-C12. Saturated monocyclic cycloalkyl rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. Saturated bicyclic and polycyclic cycloalkyl rings include, for example, norbomane, [2.2.2] bicyclooctane, decahydronaphthalene and adamantane. Cycloalkyl groups can also be partially unsaturated, having one or more double or triple bonds in the ring. Representative cycloalkyl groups that are partially unsaturated include, but are not limited to, cyclobutene, cyclopentene, cyclohexene, cyclohexadiene (1,3- and 1,4-isomers), cycloheptene, cycloheptadiene, cyclooctene, cyclooctadiene (1,3-, 1,4- and 1,5-isomers), norbomene, and norbomadiene. When cycloalkyl is a saturated monocyclic C's-C's cycloalkyl, exemplary groups include, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.

[0024] “Alkoxy” refers to an alkyl group having an oxygen atom that connects the alkyl group to the point of attachment: alkyl-O-. Alkoxy groups can have any suitable number of carbon atoms, such as Ci-Ce. Alkoxy groups include, for example, methoxy, ethoxy, propoxy, iso-propoxy, butoxy, 2-butoxy, iso-butoxy, sec-butoxy, tert-butoxy, pentoxy, hexoxy, etc.

[0025] “Heterocycloalkyl” means a monocylic or bicyclic ring system having from 3 ring members to 10 ring members and from 1 to about 5 heteroatom ring vertices selected fom N, O and S. The heteroatoms can also be oxidized, such as, but not limited to, -S(O)- and -S(O)2-. Heterocycloalkyl moieties can be saturated or include one double bond. For example, heterocycloalkyl groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, morpholino, pyrrolidinyl, pyrrolinyl, imidazolidinyl, pyrazolidinyl, pyrazolinyl, piperazinyl, and piperidinyl.

[0026] “Halogen” or “halo” refers to fluoro, chloro, bromo, or iodo.

III. Detailed Description of Embodiments

A. Methods of preventing, reducing, ameliorating, mitigating, slowing the progression and/or treating Alzheimer’s disease

[0027] Provided herein are methods of preventing, reducing, ameliorating, mitigating, slowing the progression and/or treating Alzheimer’s disease using a tight binding soluble epoxide hydrolase (sEH) inhibitor. The tight binding sEH inhibitors typically exhibit a slow dissociation rate, a long half-life of dissociation, as well as a low inhibition constant. [0028] In some aspects, provided herein are methods of preventing Alzheimer’s disease in a human subject comprising administering to the human subject a therapeutically effective amount of a tight binding soluble epoxide hydrolase (sEH) inhibitor, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 5 minutes as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay; or the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 4 nM or lower as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay.

[0029] In some aspects, provided herein are methods of reducing Alzheimer’s disease in a human subject comprising administering to the human subject a therapeutically effective amount of a tight binding soluble epoxide hydrolase (sEH) inhibitor, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 5 minutes as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay; or the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 4 nM or lower as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay.

[0030] In some aspects, provided herein are methods of ameliorating Alzheimer’s disease in a human subject comprising administering to the human subject a therapeutically effective amount of a tight binding soluble epoxide hydrolase (sEH) inhibitor, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 5 minutes as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay; or the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 4 nM or lower as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay.

[0031] In some aspects, provided herein are methods of mitigating Alzheimer’s disease in a human subject comprising administering to the human subject a therapeutically effective amount of a tight binding soluble epoxide hydrolase (sEH) inhibitor, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 5 minutes as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay; or the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 4 nM or lower as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay.

[0032] In some aspects, provided herein are methods of slowing the progression Alzheimer’s disease in a human subject comprising administering to the human subject a therapeutically effective amount of a tight binding soluble epoxide hydrolase (sEH) inhibitor, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 5 minutes as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay; or the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 4 nM or lower as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay.

[0033] In some aspects, provided herein are methods of treating Alzheimer’s disease in a human subject comprising administering to the human subject a therapeutically effective amount of a tight binding soluble epoxide hydrolase (sEH) inhibitor, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 5 minutes as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay; or the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 4 nM or lower as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay.

[0034] Soluble epoxide hydrolase inhibitors with slow dissociation rates from human sEH protein, having a long half-life of dissociation from human sEH protein are unexpectedly effective at preventing, reducing, ameliorating, mitigating, slowing the progression or treatment Alzheimer’s disease. Without being bound to any particular theory, it is believed that the slow dissociation rate and long half-life of dissociation from human sEH allows for the relative bioaccumulation of these inhibitors in the brain and other tissues such as liver with high levels of the target sEH protein.

[0035] The tight binding sEH inhibitor can be characterized using a variety of kinetic parameters. In some embodiments, the tight binding sEH inhibitor is characterized by its in vitro half-life of dissociation from human sEH protein. For the purposes of this disclosure, the in vitro half-life of dissociation from human sEH protein of a given compound is determined by using a Forster resonance energy transfer (FRET)-based competitive displacement assay described in Example 1.

[0036] In some embodiments, the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 minutes. In some embodiments, the tight binding sEH inhibitor has an in vitro halflife of dissociation from human sEH protein of at least 5, 7.5, 10, 12.5, 15, 17.5, or 20 minutes. In some embodiments, the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 5 minutes. In some embodiments, the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 7.5 minutes. In some embodiments, the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 10 minutes. In some embodiments, the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 12.5 minutes. In some embodiments, the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 15 minutes. In some embodiments, the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 17.5 minutes. In some embodiments, the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 20 minutes.

[0037] In some embodiments, the tight binding sEH inhibitor is characterized by its in vitro inhibition constant (Ki) on human sEH protein. For the purposes of this disclosure, the in vitro inhibition constant (Ki) on human sEH protein of a given compound is determined by using a Forster resonance energy transfer (FRET)-based competitive displacement assay described in Example 1. [0038] In some embodiments, the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 4, 3.5, 3, 2.5, 2, 1.5, 1, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15, 0.1, or 0.05 nM. In some embodiments, the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 2 nM, 1 nM, 500 pM, 250 pM, 100 pM, 50 pM or lower. In some embodiments, the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 2 nM or lower. In some embodiments, the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 1 nM or lower. In some embodiments, the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 500 pM or lower. In some embodiments, the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 250 pM or lower. In some embodiments, the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 100 pM or lower. In some embodiments, the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 50 pM or lower.

B. Soluble epoxide hydrolase (sEH) inhibitors

[0039] As described herein, soluble epoxide hydrolase inhibitors with slow dissociation rates and long dissociation half-lives from human sEH protein provide advantageous properties. That is, tight binding compounds with low blood-brain barrier permeability unexpectedly can prevent, reduce, ameliorate, mitigate, slow the progression, or treat Alzheimer’s disease in a human subject.

[0040] Suitable tight binding soluble epoxide hydrolase (sEH) inhibitors include, but are not necessarily limited to, compounds of Formula I

wherein

X¹ is C(O) or S(O)₂;

Y is CH or N; Z is CH₂ or NH; each R¹ is independently selected from the group consisting of H, halogen, Ci-6 alkyl, Ci-6 alkoxy, Ci-6 haloalkyl, Ci-6 haloalkoxy, -O-aryl, 5- to 6- membered heterocycloalkyl having 1 to 3 heteroatoms as ring vertices selected from N, O, and S, -OH, -NO₂, and - C(O)OR³, wherein at least one R¹ is other than H;

R² is selected from the group consisting of Ci-6 alkyl, Ci-6 haloalkyl, Ci-6 alkoxy, Ci-6 haloalkoxy, -X²-C3-6 cycloalkyl, -X²-3- to 6-membered heterocycloalkyl having 1 to 3 heteroatoms as ring vertices selected from N, O, and S, and -X²-5- to 6- membered heteroaryl having 1 to 3 heteroatom as ring vertices selected from N, O, and S, wherein R² is optionally substituted with from 1 to 3 substituents selected from the group consisting of C1-4 alkyl, C1-4 haloalkyl, hydroxyl, -C(O)OR^a, and -Ci-4-alkylene- C(O)OR^a;

X² is selected from a bond and C1-3 alkylene

R³ and R^a are each independently H or Ci-6 alkyl; and subscript n is an integer from 1 to 5.

[0041] In some embodiments, X is C(O), Y is CH, and Z is NH.

[0042] In some embodiments, each R¹ is independently selected from the group consisting of

H, halogen, Ci-6 alkyl, Ci-6 alkoxy, Ci-6 haloalkyl, and Ci-6 haloalkoxy.

[0043] In some embodiments, each R¹ is independently selected from the group consisting of halogen, C1-3 alkyl, C1-3 alkoxy, C1-3 haloalkyl, and C1-3 haloalkoxy.

[0044] In some embodiments, each R¹ is independently selected from the group consisting of fluoro, bromo, chloro, -OCF3, and -CF3.

[0045] In some embodiments, R² is Ci-6 alkyl optionally substituted with from 1 to 3 substituents selected from C1-4 haloalkyl and hydroxyl.

[0046] In some embodiments, R² is -X²-C3-6 cycloalkyl optionally substituted with from 1 to 3 substituents selected from C1-4 alkyl, C1-4 haloalkyl, hydroxyl, -C(O)OR^a, and -Ci-4-alkylene- C(O)OR^a. [0047] In some embodiments, R² is selected from the group consisting of cyclopropyl, cyclohexyl, and cyclohexylmethyl optionally substituted with from 1 to 2 substituents selected from Ci-4 alkyl, Ci-4 haloalkyl, hydroxyl, -C(O)OH.

[0048] In some embodiments, R² is X²-3- to 6-membered heterocycloalkyl having 1 to 3 heteroatoms as ring vertices selected from N, O, and S optionally substituted with from 1 to 3 substituents selected from Ci-4 alkyl, Ci-4 haloalkyl, and hydroxyl, wherein X² is a bond.

[0049] In some embodiments, R² is selected from the group consisting of tetrahydropyranyl, tetrahydrofuranyl, dihydrofuranyl, and morpholinyl optionally substituted with from 1 to 3 substituents selected from Ci-4 alkyl, Ci-4 haloalkyl, and hydroxyl

[0050] In some embodiments, R² is R² is -X²-5- to 6- membered heteroaryl having 1 to 3 heteroatom as ring vertices selected from N, O, and S optionally substituted with from 1 to 3 substituents selected from Ci-4 alkyl, Ci-4 haloalkyl and hydroxyl, wherein X² is a bond.

[0051] In some embodiments, R² is pyrrolyl or furanyl, optionally substituted with from 1 to 3 substituents selected from Ci-4 alkyl, Ci-4 haloalkyl and hydroxyl.

[0052] In some embodiments, subscript n is 1. In some embodiments, subscript n is 2. In some embodiments, subscript n is 3. In some embodiments, subscript n is 4. In some embodiments, subscript n is 5.

[0053] In some embodiments, the tight binding sEH inhibitor has the structure of a compound in Table 1.

[0054] In some embodiments, the tight binding sEH inhibitor is selected from the group consisting of

[0055] In some embodiments, the tight binding sEH inhibitor is s

[0056] In some embodiments, the tight binding sEH inhibitor is selected from the group consisting of

[0057] In some embodiments, the tight binding sEH inhibitor is selected from the group consisting of

[0058] In some embodiments, the tight binding sEH inhibitor is selected from the group consisting of

[0059] The compounds of formula (I) can be prepared according to WO2015/148954, the entirety of which is incorporated herein by reference for all purpose. In some embodiments, the tight binding soluble epoxide hydrolase (sEH) inhibitor is a compound, formula, for subformula described in WO2015/148954.

[0060] The compounds of formula (I) may exist as salts. The compounds of formula (I) includes such salts. Examples of applicable salt forms include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, maleates, acetates, citrates, fumarates, tartrates (eg (+)- tartrates, (-)-tartrates or mixtures thereof including racemic mixtures, succinates, benzoates and salts with amino acids such as glutamic acid. These salts may be prepared by methods known to those skilled in art. Also included are base addition salts such as sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the compounds of formula (I) 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 acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived 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. Certain specific compounds of the compounds of formula (I) contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0061] Certain compounds of the compounds of formula (I) 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 compounds of formula (I). Certain compounds of the compounds of formula (I) may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the compounds of formula (I) and are intended to be within the scope of the compounds of formula (I).

C. Combination Therapy

[0062] Also contemplated herein are combination therapy approaches where the compounds of Formula I are administered in combination with an additional agent. Typically, the additional agent is an additional Alzheimer’s drug, a nonsteroidial anti-inflammatory drug (nsaid or coxib), or omega-3 fatty acids.

[0063] Such agents include: acetylcholine esterase inhibitors such as tacrine (tetrahydroaminoacridine, marketed as COGNEX), donepezil hydrochloride, (marketed as Aricepts and rivastigmine (marketed as Exelon) ; gamma-secretase inhibitors; anti-inflammatory agents such as cyclooxygenase I and II inhibitors; anti-oxidants such as Vitamin E and ginkolides; omega-3 fatty acids, immunological approaches, such as, for example, immunization with A beta peptide or administration of anti-A beta peptide antibodies; statins; and direct or indirect neurotropic agents such as Cerebrolysin, AIT-082 (Emilieu, 2000, Arch. Neurol. 57: 454), and other neurotropic agents of the future.

[0064] In some embodiments the additional agent is selected from the group consisting of donepezil, galantamine, rivastigmine, memantine, or a combination thereof.

D. Formulation and Administration

[0065] In some embodiments, the tight binding soluble epoxide hydrolase inhibitor is administered as a monotherapy.

[0066] Pharmaceutical compositions or medicaments comprising a tight binding soluble epoxide hydrolase inhibitor can be administered to a subject at a therapeutically effective dose. In some embodiments, the pharmaceutical composition or medicament comprising a tight binding soluble epoxide hydrolase inhibitor is administered to a subject in an amount sufficient to elicit an effective therapeutic response in the subject. In some embodiments, the pharmaceutical composition or medicament comprising a tight binding soluble epoxide hydrolase inhibitor can be administered to a subject at a therapeutically effective dose.

[0067] The tight binding soluble epoxide hydrolase inhibitor can be prepared and administered in a wide variety of oral, parenteral and topical dosage forms. In some embodiments, the tight binding soluble epoxide hydrolase inhibitor is administered orally or by injection, that is, intravenously, intramuscularly, intracutaneously, subcutaneously, intraduodenally, or intraperitoneally. In some embodiments, the tight binding soluble epoxide hydrolase inhibitor is administered by inhalation, for example, intranasally. In some embodiments, the tight binding soluble epoxide hydrolase inhibitor is administered transdermally.

[0068] In some embodiments of the compositions, tight binding soluble epoxide hydrolase inhibitor is co-administered with the additional agent (e.g., donepezil, galantamine, etc.).

[0069] The tight binding soluble epoxide hydrolase inhibitor and the additional agent (e.g., donepezil, galantamine, etc.) independently can be prepared and administered in a wide variety of oral, parenteral and topical dosage forms. Typically, the additional agent is administered using the prescribed administration route. In some embodiments, the tight binding soluble epoxide hydrolase inhibitor and the additional agent can be administered via the same or different routes of administration. In varying embodiments, the tight binding soluble epoxide hydrolase inhibitor and the additional agent independently can be administered orally, by injection, that is, intravenously, intramuscularly, intracutaneously, subcutaneously, intraduodenally, or intraperitoneally. In some embodiments, the tight binding soluble epoxide hydrolase inhibitor and the additional agent is administered by inhalation, for example, intranasally. In some embodiments, the tight binding soluble epoxide hydrolase inhibitor and the additional agent is administered transdermally.

[0070] Furthermore, the tight binding soluble epoxide hydrolase inhibitor and the optional additional agent (e.g., donepezil, galantamine, etc.) can be co-formulated in a single composition or can be formulated for separate co- administration. Accordingly, in some embodiments, the methods contemplate administration of compositions comprising a pharmaceutically acceptable carrier or excipient, a tight binding soluble epoxide hydrolase inhibitor, and optionally the additional agent.

[0071] For preparing the 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.

[0072] 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, sodium 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. [0073] 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.

[0074] 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.

[0075] 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, polyethylene glycols and other well-known suspending agents.

[0076] 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.

[0077] 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 biochemical 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 of sEHI by replacing the active ingredient or ingredient with an sEHI, with or without EETs. 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.

[0078] Whatever the form in which the agents that inhibit sEH are topically administered (that is, whether by solid, liquid, spray, etc.), in various embodiments they are administered at a therapeutically effective dosage of about 0.01 mg to 10 mg per 10 cm². An exemplary therapeutically effective dose for systemic administration of an inhibitor of sEH is from about 0.1 pg/kg to about 100 mg/kg, e.g., about 0.001 mg/kg to about 10 mg/kg, e.g., about 0.01 mg/kg to about 1.0 mg/kg, body weight of the mammal. In various embodiments, dose and frequency of administration of an sEH inhibitor are selected to produce plasma concentrations within the range of 2.5 pM and 30 nM.

[0079] Tight binding soluble epoxide hydrolase inhibitor can be introduced into the bowel by use of a suppository. As is known in the art, suppositories are solid compositions of various sizes and shapes intended for introduction into body cavities. Typically, the suppository comprises a medication, which is released into the immediate area from the suppository. Typically, suppositories are made using a fatty base, such as cocoa butter, that melts at body temperature, or a water-soluble or miscible base, such as glycerinated gelatin or polyethylene glycol.

[0080] 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 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.

[0081] 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.

[0082] A therapeutically effective amount or a sub-therapeutic amount of the tight binding soluble epoxide hydrolase inhibitor can be co-administered with the additional agent. 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 therapeutically effective dose is from about 0.1 pg/kg to about 100 mg/kg, e.g., about 0.001 mg/kg to about 10 mg/kg, e.g., about 0.01 mg/kg to about 1.0 mg/kg, body weight of the mammal. Determination of an effective amount is well within the capability of those skilled in the art.

[0083] Generally, an efficacious or effective amount of a combination of one or more agents 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 one or more agents 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), 69^th Edition, 2015 and 70^th Edition, 2016, PDR Network; in Remington: The Science and Practice of Pharmacy, 21^st 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.

[0084] In some embodiments, prior to administration of the tight binding soluble epoxide hydrolase inhibitor, the individual to be treated has been previously diagnosed as having Alzheimer’s disease. [0085] In some embodiments, the therapeutically effective amount of a tight binding soluble epoxide hydrolase (sEH) inhibitor is a total daily dosage of about 2 mg to 20 mg (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg/day). In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 2 to 7 mg, 8 to 14 mg, 15 to 20 mg, 5 to 15 mg or 7.5 to 12.5 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 2 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 3 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 4 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 5 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 6 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 7 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 8 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 9 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 10 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 11 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 12 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 13 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 14 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 15 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 16 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 17 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 18 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 19 mg. In some embodiments, the total daily dosage of a tight binding soluble epoxide hydrolase (sEH) inhibitor is about 20 mg. In some embodiments, the tight binding soluble epoxide hydrolase (sEH) inhibitor is Compound 1.004. In some embodiments, the tight binding soluble epoxide hydrolase (sEH) inhibitor is Compound 1.010. In some embodiments, the above dosages are those administered to humans.

[0086] In some embodiments, a tight binding soluble epoxide hydrolase (sEH) inhibitor is administered orally. In some embodiments, a tight binding soluble epoxide hydrolase (sEH) inhibitor is administered daily in single, divided, or continuous doses. In some embodiments, a tight binding soluble epoxide hydrolase (sEH) inhibitor is administered twice daily. In some embodiments, a tight binding soluble epoxide hydrolase (sEH) inhibitor is administered three times daily. In some embodiments, the tight binding soluble epoxide hydrolase (sEH) inhibitor is Compound 1.004. In some embodiments, the tight binding soluble epoxide hydrolase (sEH) inhibitor is Compound 1.010.

E. Kits

[0087] Further provided herein are kits. In varying embodiments, the kits comprise one or more tight binding soluble epoxide hydrolase inhibitors. In some embodiments, the kits further comprise one or more additional agents described herein. Embodiments of the tight binding soluble epoxide hydrolase inhibitor and embodiments of the additional agent(s) are as described above and herein. Embodiments of formulations of the agents are as described above and herein. In varying embodiments, the tight binding soluble epoxide hydrolase inhibitor and the additional agent(s) can be co-formulated for administration as a single composition. In some embodiments, the tight binding soluble epoxide hydrolase inhibitor and the additional agent(s) are formulated for separate administration, e.g. , via the same or different route of administration. In varying embodiments, one or both the tight binding soluble epoxide hydrolase inhibitor and the additional agent(s) are provided in unitary dosages in the kits.

[0088] Some of the kits described herein include a label describing a method of administering one or more tight binding soluble epoxide hydrolase inhibitor and/or one or more additional therapeutic agents described herein.

IV. Examples

[0089] The following examples are provided to illustrate by not limit the claimed invention. Example 1: Forster resonance energy transfer (FRET)-based competitive displacement assay

[0090] This example describes how the in vitro half-life of dissociation from human sEH protein for a given compound is determined. The assay is performed as described in Lee et al. Anal Biochem. 2013; 434(2): 259-268 and Lee et al. J. Med. Chem. 2014, 57, 16, 7016-7030.

[0091] In this assay, the pre-incubated recombinant sEH- sEH inhibitor complex was diluted in the buffer in the presence of high concentration of fluorescent ligand. Upon dilution, the sEH inhibitor was dissociated from sEH and the vacant sEH was bound by fluorescent ligand which led to fluorescent increase over time. By fitting the obtained fluorescent enhancement curve with single exponential rise to maximum equation, the toff was calculated and the In (2) over k_Off resulted a ti/2 of inhibitor on sEH.

[0092] In vitro half-life of dissociation from human sEH protein for certain compounds are displayed in Table 1, below.

Table 1 : in vitro half-life of dissociation from human sEH protein

^a ti/2 defined as the time required for half of the drug being dissociated from the enzyme based on the fluorescence signals.

[0093] The referenced Lee papers, above, also describe how to determine and calculate Ki. Inhibition constant (Ki) values. In short, the pre-incubated soluble epoxide hydrolase (sEH)- fluorescent ligand complex was titrated with increasing concentration of selected sEH inhibitor.

The fluorescent ligand was displaced by inhibitors and fluorescence decreased. The fluorescence intensity was plotted against the concentration of the inhibitor. By fitting this curve with cubic fit equation using program Sigmaplot 8.0 as described in Lee et al. 2013 and 2014, the Ki can be calculated. [0094] Inhibition constant (Ki) values for select compounds with human sEH protein are displayed in Table 2, below.

Table 2: Inhibition Constant (Ki) human sEH protein

Example 2: Surprising in vivo blood-brain barrier bioaccumulation in mice despite poor predicted permability [0095] Predictive models for the above-described compounds indicate that these compound are expected to poorly penetrate the blood-brain barrier. See, for example, Example 3. Surprisingly, mice dosed with 1 mg/kg of the sEH inhibitors referenced below by oral gavage exhibit measurable blood-brain barrier penetration beyond initial expectation. [0096] Table 3, below, reports the amount of sEH inhibitor measured in the blood pre-dose (0 min) and a 1 hour time point, the concentration of sEH inhibitor measured in brain after 1 hour, and the relative amounts in the blood v. the brain after 1 hour (reported as brain/blood ratio).

Table 3: sEHi Concentrations in Blood and Brain tissues after administration to mice

[0097] Blood and brain sEHi concentrations were determined using suitable extraction methods for each sample type followed by LC-MS/MS analysis.

[0098] It is believed that the uniquely slow dissociation rate and low Ki of the described sEH inhibitors results in a surprising bioaccumulation of these compounds in the brain, beyond what is expected using predictive modeling methods. [0099] From the table above, it appears that the longer half-life of dissociation and lower inhibition constant of Compound 1.004 (ti/2 is 23.3 min and k_Off is 4.96 x 10’⁴ s’¹ in murine sEH protein) unexpectedly provides (on average) a more than five-fold higher brain/blood ratio as compared to Compound 1.010 (ti/2 is 3.7 min and k_Off is 8.43 x 10’⁴ s’¹ in murine sEH protein) 1 hour after dosing. Based on the currently discovered surprising effect in mice, unexpectedly higher brain/blood ratios are also expected in humans. Example 3: Computational Prediction for Blood-Brain Barrier (BBB) penetration using a Central Nervous System Multiparameter Optimization (CNS MPO) Approach

[0100] This example describes how the CNS MPO scores for a given compound is determined. The in silico method was performed as described in Wager et al. ACS Chem. Neurosci. 2010; 1 : 435 449. [0101] The prediction for the BBB penetration of sEH inhibitors was guided by an in silico approach called a CNS multiparameter optimization (MPO), which scores small molecules based on 6 fundamental physicochemical parameters that are important for understood CNS penetration: calculated logP (ClogP), calculated logD at pH 7.4 (ClogD), molecular weight (MW), topological polar surface area (TPSA), hydrogen bonding donors (HBDs) and acidity (pKa). The CNS MPO score ranges from 1 to 6 (6 being the best) and a CNS MPO score >4.5 is considered to be desirable for a CNS drug.

[0102] CNS MPO scores for certain compounds are displayed in Table 3, below. Overall, these compounds generally have a moderate or low expected blood-brain barrier permeability.

Table 4: in vitro half-life of dissociation from human sEH protein

[0103] 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.

Claims

WHAT IS CLAIMED IS:

1. A method of preventing, reducing, ameliorating, mitigating, slowing the progression and/or treating Alzheimer’s disease in a human subject comprising administering to the human subject a therapeutically effective amount of a tight binding soluble epoxide hydrolase (sEH) inhibitor, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 5 minutes as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay; or the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 4 nM or lower as determined by a Forster resonance energy transfer (FRET)-based competitive displacement assay.

2. The method of claim 1 , wherein the tight binding sEH inhibitor has the structure of Formula I:

wherein

X¹ is C(O) or S(O)₂;

Y is CH or N;

Z is CH₂ orNH; each R¹ is independently selected from the group consisting of H, halogen, Ci-6 alkyl, Ci-6 alkoxy, Ci-6 haloalkyl, Ci-6 haloalkoxy, -O-aryl, 5- to 6- membered heterocycloalkyl having 1 to 3 heteroatoms as ring vertices selected from N, O, and S, -OH, -NO₂, and - C(O)OR³, wherein at least one R¹ is other than H;

R² is selected from the group consisting of Ci-6 alkyl, Ci-6 haloalkyl, Ci-6 alkoxy, Ci-6 haloalkoxy, -X²-C3-6 cycloalkyl, -X²-3- to 6-membered heterocycloalkyl having 1 to 3 heteroatoms as ring vertices selected from N, O, and S, and -X²-5- to 6- membered heteroaryl having 1 to 3 heteroatom as ring vertices selected from N, O, and S, wherein R² is optionally substituted with from 1 to 3 substituents selected from the group consisting of Ci-4 alkyl, Ci-4 haloalkyl, hydroxyl, -C(O)OR^a, and -Ci-4-alkylene- C(O)OR^a;

X² is selected from a bond and C1-3 alkylene

3. The method of claim 2, wherein X is C(O), Y is CH, and Z is NH.

4. The method of claim 2 or claim 3, wherein each R¹ is independently selected from the group consisting of H, halogen, Ci-6 alkyl, Ci-6 alkoxy, Ci-6 haloalkyl, and Ci-6 haloalkoxy.

5. The method of claim 4, wherein each R¹ is independently selected from the group consisting of halogen, C1-3 alkyl, C1-3 alkoxy, C1-3 haloalkyl, and C1-3 haloalkoxy.

6. The method of claim 5, wherein each R¹ is independently selected from the group consisting of fluoro, bromo, chloro, -OCF3, and -CF3.

7. The method of any one of claims 2 to 6, wherein R² is Ci-6 alkyl optionally substituted with from 1 to 3 substituents selected from C1-4 haloalkyl and hydroxyl.

8. The method of any one of claims 2 to 6, wherein R² is -X²-C3-6 cycloalkyl optionally substituted with from 1 to 3 substituents selected from C1-4 alkyl, C1-4 haloalkyl, hydroxyl, -C(O)OR^a, and -Ci-4-alkylene- C(O)OR^a.

9. The method of claim 8, wherein R² is selected from the group consisting of cyclopropyl, cyclohexyl, and cyclohexylmethyl optionally substituted with from 1 to 2 substituents selected from Ci-4 alkyl, Ci-4 haloalkyl, hydroxyl, and -C(O)OH.

10. The method of any one of claims 2 to 6, wherein R² is X²-3- to 6- membered heterocycloalkyl having 1 to 3 heteroatoms as ring vertices selected from N, O, and S optionally substituted with from 1 to 3 substituents selected from C1-4 alkyl, C1-4 haloalkyl, and hydroxyl, wherein X² is a bond.

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11. The method of claim 10, wherein R² is selected from the group consisting of tetrahydropyranyl, tetrahydrofuranyl, dihydrofuranyl, and morpholinyl optionally substituted with from 1 to 3 substituents selected from Ci-4 alkyl, Ci-4 haloalkyl, and hydroxyl.

12. The method of any one of claims 2 to 6, wherein R² is -X²-5- to 6- membered heteroaryl having 1 to 3 heteroatom as ring vertices selected from N, O, and S optionally substituted with from 1 to 3 substituents selected from Ci-4 alkyl, Ci-4 haloalkyl and hydroxyl, wherein X² is a bond.

13. The method of claim 12, wherein R² is pyrrolyl or furanyl, optionally substituted with from 1 to 3 substituents selected from Ci-4 alkyl, Ci-4 haloalkyl and hydroxyl.

14. The method of any one of claims 1 to 13, wherein the subscript n is 1.

15. The method of any one of claims 1 to 13, wherein the subscript n is 2.

16. The method of claim 1 , wherein the tight binding sEH inhibitor has the structure of a compound in Table 1.

17. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 7.5 minutes.

18. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 10 minutes.

19. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 12.5 minutes.

20. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 15 minutes.

21. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 17.5 minutes.

22. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro half-life of dissociation from human sEH protein of at least 20 minutes.

23. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 2 nM or lower.

24. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 1 nM or lower.

25. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 500 pM or lower.

26. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 250 pM or lower.

27. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 100 pM or lower.

28. The method of any one of claims 1 to 16, wherein the tight binding sEH inhibitor has an in vitro inhibition constant (Ki) on human sEH protein of about 50 pM or lower.

29. A method of any one of claims 1 to 28, wherein said therapeutically effective amount of the tight binding sEH inhibitor is a total daily dosage in human of from about 2 to 20 mg per day.

30. A method of claim 29, wherein said total daily dosage in human is from about 5 to 15 mg per day.

31. A method of claim 29, wherein said total daily dosage in human is from about 7.5 to 12.5 mg per day.

32. A method of claim 29, wherein said total daily dosage in human is from about 2 mg per day.

33. A method of claim 29, wherein said total daily dosage in human is from about 6 mg per day.

34. A method of claim 29, wherein said total daily dosage in human is from about 10 mg per day.

35. The method of any one of claims 1 to 34, wherein the tight binding sEH inhibitor is administered orally.

36. The method of any one of claims 1 to 35, wherein the tight binding sEH inhibitor is administered once daily.

37. The method of any one of claims 1 to 35, wherein the tight binding sEH inhibitor is administered twice daily.

38. The method of any one of claims 1 to 37, further comprising administering an additional therapeutic agent.

39. The method of claim 38, where the additional therapeutic agent is selected from the group consisting of donepezil, galantamine, rivastigmine, memantine, or a combination thereof.

40. A method of preventing, reducing, ameliorating, mitigating, slowing the progression and/or treating Alzheimer’s disease in a human subject comprising administering to the human subject a compound having the chemical structure

41. A method of claim 40, wherein said therapeutically effective amount of the tight binding sEH inhibitor is a total daily dosage in human of from about 2 to 20 mg per day.

42. A method of claim 41, wherein said total daily dosage in human is from about 5 to 15 mg per day.

43. A method of claim 41, wherein said total daily dosage in human is from about 7.5 to 12.5 mg per day.

44. The method of claim 41, wherein said total daily dosage in human is from about 2 mg per day.

45. The method of claim 41, wherein said total daily dosage in human is from about 6 mg per day.

46. The method of claim 41, wherein said total daily dosage in human is from about 10 mg per day.

47. The method of claim 40, wherein the compound is administered orally.

48. The method of claim 40, wherein the compound is administered once daily.

49. The method of claim 40, wherein the compound is administered twice daily.

39