Combination of thyromimetics with fatty acid amide hydrolase (FAAH)-cleavable prodrugs and peripherally restricted FAAH inhibitors

Abstract

Provided herein are fatty acid amide hydrolase (FAAH) cleavable prodrugs of thyromimetics, and pharmaceutical compositions comprising these compounds, at least one pharma- ceutically acceptable excipient, and a peripherally restricted FAAH inhibitor.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 185,254, filed May 6, 2021, and U.S. Provisional Patent Application No. 63 / 274,856, filed November 2, 2021, each of which is incorporated by reference in its entirety. [Background technology]

[0002] The blood-brain barrier is composed of tightly interconnected endothelial cells that restrict the passage of pathogens and certain types of small and large molecules from the blood to the brain. This important protective function also limits the diffusion of therapeutic drugs into the brain, presenting a major challenge for the development of novel medicines for CNS diseases. Summary of the Invention

[0003] In one aspect herein, a compound of formula (I):

[0004] [ka] A fatty acid amide hydrolase (FAAH) cleavable prodrug of the formula: R 1 and R 2 are each independently hydrogen, -OR 5 , -NR 5 R 6 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, phenyl, and -C1-C6 alkyl-phenyl, wherein C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, phenyl, and -C1-C6 alkyl-phenyl are selected from halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5optionally replaced by one or more of R 3 and R 4 is independently selected from -F, -Cl, -Br, and -I; R 5 and R 6 is independently selected from hydrogen and C1-C6 alkyl; R 7 and R 8 is independently selected from hydrogen, -F, -Cl, -Br, and -I, and a pharma- ceutically acceptable excipient, or a pharma- ceutically acceptable salt or solvate thereof, further comprising a peripherally restricted FAAH inhibitor.

[0005] In some embodiments, R 7 is hydrogen. In some embodiments, R 7 is -F. In some embodiments, R 8 is hydrogen. In some embodiments, R 8 is -F.

[0006] In another aspect herein, a compound of formula (I):

[0007] [ka] A fatty acid amide hydrolase (FAAH) cleavable prodrug of the formula: R 1 and R 2 are each independently hydrogen, -OR 5 , -NR 5 R 6, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, phenyl, and -C1-C6 alkyl-phenyl, wherein C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, phenyl, and -C1-C6 alkyl-phenyl are selected from halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 optionally replaced by one or more of R 3 and R 4 is independently selected from -F, -Cl, -Br, and -I; R 5 and R 6 is independently selected from hydrogen and C1-C6 alkyl, and a pharma- ceutically acceptable excipient, or a pharma- ceutically acceptable salt or solvate thereof, further comprising a peripherally restricted FAAH inhibitor.

[0008] In some embodiments, R 1 is hydrogen. In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2is C1-C6 alkyl substituted with one or more -OH. In some embodiments, R 2 is C-C alkyl substituted with one or more halo. In some embodiments, R 2 is unsubstituted C1-C6 alkyl. In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R is phenyl optionally substituted with one or more of 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 3 and R 4 is independently selected from -F, -Cl, and -Br. 3 and R 4 and R are both -Br. 3 and R 4 and R are both -Br. 3 and R 4 and R are both -Cl. 3 and R 4 are both -F.

[0009] In some embodiments, the peripherally restricted FAAH inhibitor is ASP-3652.

[0010] In another aspect, a method for treating a CNS disease or disorder in a patient in need of such treatment comprises administering to the patient a therapeutically effective amount of a pharmaceutical composition as described herein.In some embodiments, the CNS disease or disorder is acute disseminated encephalomyelitis (ADEM), acute hemorrhagic leukoencephalitis (AHL or AHLE), adult Refsum disease, infantile Refsum disease, Alexander disease, Alzheimer's disease, Baro concentric sclerosis, Canavan disease, central pontine myelinolysis (CPM), cerebral palsy, cerebrotendinous xanthomatosis, chronic inflammatory demyelinating polyneuropathy (CIDP), Devic's syndrome, diffuse spinal cord fragmentation sclerosis, encephalomyelitis, Guillain-Barre syndrome, idiopathic inflammatory demyelinating disease (HDD), Krabbe disease, The CNS disease or disorder is selected from Leber's hereditary optic atrophy, leukodystrophy, Marburg multiple sclerosis, Marchiafava-Bienami disease, metachromatic leukodystrophy (MLD), multifocal motor neuropathy (MMN), multiple sclerosis (MS), paraproteinemic demyelinating polyneuropathy, Pelizaeus-Merzbach disease (PMD), progressive multifocal leukoencephalopathy (PML), tropical spastic paraparesis (TSP), X-linked adrenoleukodystrophy (X-ALD, ALO, or X-linked ALO), and Zellweger syndrome. In some embodiments, the CNS disease or disorder is selected from multiple sclerosis and X-linked adrenoleukodystrophy. [Brief description of the drawings]

[0011] The novel features of the present disclosure are set forth with particularity in the appended claims. The features and advantages of the present disclosure will be better understood by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings.

[0012] [Figure 1] FIG. 2 shows that LL-341070A, an active metabolite of the LL-341070 prodrug, enhanced oligodendrocyte differentiation in vitro in an oligodendrocyte precursor cell assay. [Diagram 2]FIG. 1: Thyromimetic treatment enhances 24-OHC synthesis in vivo in rat brain following cuprizone-induced demyelination. [Diagram 3] FIG. 1 shows that TRβ target engagement in the brain is demonstrated by increased expression of T3-responsive target genes in vivo. [Figure 4] FIG. 1 shows brain and plasma concentrations of LL-341070 after 21 days of repeated dosing, measured 4 hours after the final dose. [Diagram 5] FIG. 2 shows that LL-341070 improves in vivo clinical scores in a mouse prophylactic experimental autoimmune encephalitis (EAE) model. [Figure 6] FIG. 1 depicts FAAH expression and specific activity across species and tissue types. [Figure 7] FIG. 1 depicts concentrations of ABX-002A in the brain, liver, kidneys, lungs, and heart measured 1 hour after SC administration of 30 different prodrugs of ABX-002A. [Figure 8] FIG. 1 depicts plasma, liver, and brain concentrations following ABX-002 prodrug treatment with or without peripheral or global FAAH inhibitors. [Figure 9A] FIG. 13 depicts T3 target gene transfer in brain vs. liver following a single dose of ABX-002A. [Figure 9B] FIG. 13 depicts T3 target gene transfer in brain vs. liver following a single dose of ABX-002. [Figure 9C] FIG. 13. T3 target gene transfer in brain vs. liver following a single dose of ABX-002A and a FAAH inhibitor. [Figure 10A] FIG. 1 depicts gene expression in the brain and liver following administration of ABX-002A, and the effect on T4. [Figure 10B] FIG. 1 depicts gene expression in the brain and liver following administration of ABX-002, and the effect on T4. [Figure 10C]FIG. 1 depicts gene expression in brain and liver following administration of ABX-002 and a peripheral FAAH inhibitor, and the effect on T4. [Figure 10D] FIG. 1 shows gene expression in brain and liver following administration of ABX-002 and a global FAAH inhibitor, and the effect on T4. [Figure 11A] FIG. 1 depicts T4 inhibition as a function of ABX-002 dose in mice or non-human primates (NHPs). [Figure 11B] FIG. 13 represents T4 inhibition as a function of plasma ABX-002 prodrug AUC in mice or non-human primates (NHPs). [Figure 11C] FIG. 1 represents T4 inhibition as a function of plasma ABX-002A active metabolite AUC in mice or non-human primates (NHPs). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] Fatty acid hydrolases (FAAHs) are integral membrane serine hydrolases that can degrade the fatty acid amide family of signaling lipids to hydrolyze selected amide prodrugs. FAAHs are highly conserved among species and expressed to various degrees in many tissues, including the central nervous system (CNS). Selected carboxylic acids can be converted to highly permeable amide prodrugs that can then cross the blood-brain barrier, where they can be converted to active molecules by the action of FAAH on the prodrug. This results in higher amounts of carboxylic acids being delivered to the CNS compared to parent dosing alone. However, peripherally expressed FAAHs can simultaneously hydrolyze the prodrug, resulting in a significant amount of non-productive prodrug conversion. Coadministration of peripherally restricted FAAH inhibitors with CNS-permeable FAAH-convertible prodrugs improves the selectivity of prodrug delivery to the CNS. In addition, exposure of the parent molecule in plasma and peripheral tissues is less than that observed when the prodrug is administered alone.

[0014] Candidates for clinical development may be selected from the compounds disclosed herein based on their degree of in vitro FAAH-mediated hydrolysis, in vitro plasma stability, in vivo tissue distribution, in vitro target selectivity, in vitro target potency, target gene expression, in vivo pharmacological efficacy, or drug-like (rule-of-5 compliant) physiochemical properties, or a combination thereof.

[0015] Specific Terms The singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, a reference to "a drug" includes a reference to one or more of such drugs, and a reference to "an excipient" includes a reference to one or more of such excipients. When ranges are used herein, it is intended that all combinations and subcombinations of ranges and specific embodiments therein are included. When referring to a number or range of numbers, the term "about" means that the number or range of numbers referred to is an approximation within experimental variation (or within statistical experimental error), such that the number or range of numbers varies between 1% and 15% of the stated number or range of numbers.

[0016] The terms "formulation" and "composition" are used interchangeably as used herein and refer to a mixture of two or more compounds, elements, or molecules. In some embodiments, the terms "formulation" and "composition" may also be used to refer to a mixture of one or more active agents with a carrier or other excipient.

[0017] The terms "active agent," "active pharmaceutical agent," "drug," "active ingredient," and variations thereof, are used interchangeably to refer to an agent or substance that has a measurable, specific or selected biological activity when administered to a subject in a significant or effective amount.

[0018] "Pharmaceutically acceptable salt" includes both acid and base addition salts. A pharmaceutically acceptable salt of any one of the compounds described herein is intended to include all pharmaceutically appropriate salt forms. Preferred pharmaceutically acceptable salts of the compounds described herein are pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0019] "Pharmaceutically acceptable acid addition salts" refer to salts that retain the biological effectiveness and properties of the free bases, and are not biologically or otherwise undesirable, and are formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, etc. Also included are salts formed with organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanediol acids, aromatic acids, and aliphatic and aromatic sulfonic acids, including, for example, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Thus, exemplary salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, trifluoroacetates, propionates, caprylates, isobutyrates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, mandelates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, malates, tartrates, methanesulfonates, and the like. Additionally, salts of amino acids such as arginate, gluconate, galacturonate, and the like are contemplated (see, for example, Berge SM et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 66:1-19 (1997)). Acid addition salts of basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt.

[0020] "Pharmaceutically acceptable base addition salts" refer to salts that retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared by adding an inorganic or organic base to the free acid. In some embodiments, pharma-ceutically acceptable base addition salts are formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Salts derived from organic bases include, but are not limited to, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, N,N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline, N-methylglucamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. See Berge et al., supra.

[0021] It should be understood that reference to pharmaceutically acceptable salts includes solvent addition forms (solvates). Solvates contain stoichiometric or non-stoichiometric amounts and are formed during the process of product formation or isolation with pharmaceutically acceptable solvents such as water, ethanol, methanol, methyl tert-butyl ether (MTBE), diisopropyl ether (DIPE), ethyl acetate, isopropyl acetate, isopropyl alcohol, methyl isobutyl ketone (MIBK), methyl ethyl ketone (MEK), acetone, nitromethane, tetrahydrofuran (THF), dichloromethane (DCM), dioxane, heptane, toluene, anisole, acetonitrile, etc. In one embodiment, solvates are formed with, but not limited to, Class 3 solvents. Solvent categories are defined, for example, in Impurities: Guidelines for Residual Solvents, Q3C(R3), (November 2005) by the International Conference on Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH). Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is alcohol.

[0022] The term "effective amount" or "therapeutically effective amount" as used herein refers to an amount of an administered drug or compound sufficient to alleviate to some extent one or more of the symptoms of the disease or disorder being treated. The result may be a reduction and / or alleviation of the signs, symptoms, or causes of the disease, or other desired alteration of a biological system. For example, an "effective amount" in therapeutic applications is the amount of a composition containing a compound disclosed herein that is required to produce a clinically significant reduction in the disease. An appropriate "effective" amount in any individual case may optionally be determined using techniques such as dose escalation studies.

[0023] The terms "subject," "individual," and "patient" are used interchangeably herein to refer to mammals, including, but not limited to, murines, simians, humans, farm animals, sport animals, and pets.

[0024] The term "peripherally restricted FAAH inhibitor" as used herein refers to a FAAH inhibitor that inhibits fatty acid amide hydrolase (FAAH) in the peripheral nervous system to a greater extent than in the central nervous system upon systemic administration. In some embodiments, the peripherally restricted FAAH inhibitor is 60% peripherally restricted. In some embodiments, the peripherally restricted FAAH inhibitor is 70% peripherally restricted. In some embodiments, the peripherally restricted FAAH inhibitor is 80% peripherally restricted. In some embodiments, the peripherally restricted FAAH inhibitor is 90% peripherally restricted. In some embodiments, the peripherally restricted FAAH inhibitor is 95% peripherally restricted.

[0025] target Thyroid hormone (TH) is a major signal for oligodendrocyte differentiation and myelination during development and also stimulates remyelination in adult models of multiple sclerosis (MS) (Calza L et al. Brain Res Revs 48:339-346, 2005). However, TH is not an acceptable long-term therapy because there is virtually no therapeutic window in which remyelination can be achieved while avoiding the cardiotoxicity and bone demineralization associated with chronic hyperthyroidism. Some thyroid hormone analogs are able to activate thyroid hormone-responsive genes while avoiding the associated downsides of TH by exploiting the molecular and physiological characteristics of the thyroid hormone receptor (Malm J et al. Mini Rev Med Chem 7:79-86, 2007). These receptors are expressed in two major forms with heterogeneous tissue distribution and overlapping but distinct sets of target genes (Yen PM, Physiol Rev 81:1097-1142, 2001). TRα is abundant in the heart, brain, and bone, whereas TRβ is abundant in the liver (O'Shea PJ et al., Nucl Recept Signal 4:e011, 2006).

[0026] The development of selective thyromimetics has been difficult due to the high degree of sequence homology of the thyroid hormone receptor subtypes, i.e., only one amino acid residue on the interior surface of the ligand-binding domain cavity varies between the α1 and β1 forms.

[0027] In some embodiments, the pharmaceutical compositions described herein comprise a fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (I'), where the prodrug of formula (I') is a prodrug of a TRβ agonist. In some embodiments, the pharmaceutical compositions described herein comprise a fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (I), where the prodrug of formula (I) is a prodrug of a TRβ agonist. In some embodiments, the pharmaceutical compositions described herein comprise a fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (II), where the prodrug of formula (II) is a prodrug of a TRβ agonist.

[0028] Pharmaceutical Compositions In some embodiments herein, the compound of formula (I):

[0029] [ka] A fatty acid amide hydrolase (FAAH) cleavable prodrug of the formula: R 1 and R 2 are each independently hydrogen, -OR 5 , -NR 5 R 6 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, phenyl, and -C1-C6 alkyl-phenyl, wherein C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, phenyl, and -C1-C6 alkyl-phenyl are selected from halo, cyano, -OR 5 , -NR 5 R6 , -S(O)2R 5 , or -S(O)2OR 5 optionally replaced by one or more of R 3 and R 4 is independently selected from -F, -Cl, -Br, and -I; R 5 and R 6 is independently selected from hydrogen and C1-C6 alkyl; R 7 and R 8 is independently selected from hydrogen, -F, -Cl, -Br, and -I; and a pharma- ceutically acceptable excipient, or a pharma- ceutically acceptable salt or solvate thereof, further comprising a peripherally restricted FAAH inhibitor.

[0030] In some embodiments, R 7 is hydrogen. In some embodiments, R 7 is -F. In some embodiments, R 7 is -Cl. In some embodiments, R 7 is -Br.

[0031] In some embodiments, R 8 is hydrogen. In some embodiments, R 8 is -F. In some embodiments, R 8 is -Cl. In some embodiments, R 8 is -Br.

[0032] In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is C 1-6 It is an alkyl.

[0033] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6, -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is C-C alkyl substituted with one or more halo. In some embodiments, R 2 is C1-C6 alkyl substituted with one cyano. In some embodiments, R 2 can be one or more -OR 5 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C1-C6 alkyl substituted with one or more -OH. In some embodiments, R 2 is C1-C6 alkyl substituted with one -OH. In some embodiments, R 2 is one or more -NR 5 R 6 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C1-C6 alkyl substituted with one or more -NH2. In some embodiments, R 2 is C1-C6 alkyl substituted with one -NH2. In some embodiments, R 2 is one -S(O)2R 5 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C1-C6 alkyl substituted with one -S(O)2H. In some embodiments, R 2 is one -S(O)2OR 5 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C-C alkyl substituted with one -S(O)OH. In some embodiments, R2 is unsubstituted C1-C6 alkyl. In some embodiments, R 2 is -CH3. In some embodiments, R 2 is -CH2CH3. In some embodiments, R 2 is -CH2CH2CH3.

[0034] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted C2-C6 alkenyl.

[0035] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted C2-C6 alkynyl.

[0036] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted C3-C6 cycloalkyl.

[0037] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is an unsubstituted C3-C6 heterocycloalkyl.

[0038] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R is phenyl optionally substituted with one or more of 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is phenyl substituted with one or more halo. In some embodiments, R 2 can be one or more -OR 5 In some embodiments, R 2 is phenyl substituted with one or more -OH. In some embodiments, R 2 is unsubstituted phenyl.

[0039] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted -C1-C6 alkyl-phenyl.

[0040] In some embodiments, R 2 -OR 5 In some embodiments, R 2 is -OH. In some embodiments, R 2 -NR 5 R 6 In some embodiments, R 2 is -NH2.

[0041] In some embodiments, R 2 is hydrogen.

[0042] In some embodiments, R 3 and R 4 is independently selected from -F, -Cl, and -Br. 3 and R 4 and R are both -Br. 3 and R 4 and R are both -Br. 3 and R 4 and R are both -Cl. 3 and R 4 are both -F. In some embodiments, R 3 is -Cl, R 4 In some embodiments, R 3 is -F and R 4 In some embodiments, R 3 is -F and R 4 is -Cl.

[0043] In some embodiments of the pharmaceutical compositions described herein, the fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (I') is

[0044] [ka] or a pharma- ceutically acceptable salt or solvate thereof.

[0045] In some embodiments herein, the compound of formula (I):

[0046] [ka] A fatty acid amide hydrolase (FAAH) cleavable prodrug of the formula: R 1 and R 2 are each independently hydrogen, -OR 5 , -NR 5 R 6 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, phenyl, and -C1-C6 alkyl-phenyl, wherein C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, phenyl, and -C1-C6 alkyl-phenyl are selected from halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 optionally replaced by one or more of R 3 and R 4 is independently selected from -F, -Cl, -Br, and -I; R 5 and R 6 is independently selected from hydrogen and C1-C6 alkyl, and a pharma- ceutically acceptable excipient, or a pharma- ceutically acceptable salt or solvate thereof, further comprising a peripherally restricted FAAH inhibitor.

[0047] In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is C 1-6 It is an alkyl.

[0048] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is C-C alkyl substituted with one or more halo. In some embodiments, R 2 is C1-C6 alkyl substituted with one cyano. In some embodiments, R 2 can be one or more -OR 5 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C1-C6 alkyl substituted with one or more -OH. In some embodiments, R 2 is C1-C6 alkyl substituted with one -OH. In some embodiments, R 2 is one or more -NR 5 R 6 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C1-C6 alkyl substituted with one or more -NH2. In some embodiments, R 2 is C1-C6 alkyl substituted with one -NH2. In some embodiments, R 2 is one -S(O)2R 5 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C1-C6 alkyl substituted with one -S(O)2H. In some embodiments, R 2 is one -S(O)2OR 5In some embodiments, R is a C1-C6 alkyl substituted with 2 is C-C alkyl substituted with one -S(O)OH. In some embodiments, R 2 is unsubstituted C1-C6 alkyl. In some embodiments, R 2 is -CH3. In some embodiments, R 2 is -CH2CH3. In some embodiments, R 2 is -CH2CH2CH3.

[0049] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted C2-C6 alkenyl.

[0050] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted C2-C6 alkynyl.

[0051] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted C3-C6 cycloalkyl.

[0052] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is an unsubstituted C3-C6 heterocycloalkyl.

[0053] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R is phenyl optionally substituted with one or more of 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is phenyl substituted with one or more halo. In some embodiments, R 2 can be one or more -OR 5 In some embodiments, R 2 is phenyl substituted with one or more -OH. In some embodiments, R 2 is unsubstituted phenyl.

[0054] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5In some embodiments, R 2 is unsubstituted -C1-C6 alkyl-phenyl.

[0055] In some embodiments, R 2 -OR 5 In some embodiments, R 2 is -OH. In some embodiments, R 2 -NR 5 R 6 In some embodiments, R 2 is -NH2.

[0056] In some embodiments, R 2 is hydrogen.

[0057] In some embodiments, R 3 and R 4 is independently selected from -F, -Cl, and -Br. 3 and R 4 and R are both -Br. 3 and R 4 and R are both -Br. 3 and R 4 and R are both -Cl. 3 and R 4 are both -F. In some embodiments, R 3 is -Cl, R 4 In some embodiments, R 3 is -F and R 4 In some embodiments, R 3 is -F and R 4 is -Cl.

[0058] In some embodiments of the pharmaceutical compositions described herein, the fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (I') or (I) is

[0059] [ka]

[0060] [ka]

[0061] [ka] or a pharma- ceutically acceptable salt or solvate thereof.

[0062] In some embodiments of the pharmaceutical compositions described herein, the fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (I') or (I) is

[0063] [ka]

[0064] [ka] or a pharma- ceutically acceptable salt or solvate thereof.

[0065] In some embodiments herein, the compound of formula (II):

[0066] [ka] A fatty acid amide hydrolase (FAAH) cleavable prodrug of the formula: R 1 and R 2 are each independently hydrogen, -OR 5 , -NR 5 R 6, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, phenyl, and -C1-C6 alkyl-phenyl, wherein C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, phenyl, and -C1-C6 alkyl-phenyl are selected from halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 optionally replaced by one or more of R 5 and R 6 is independently selected from hydrogen and C1-C6 alkyl, and a pharma- ceutically acceptable excipient, or a pharma- ceutically acceptable salt or solvate thereof, further comprising a peripherally restricted FAAH inhibitor that is ASP-3652.

[0067] In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is C 1-6 It is an alkyl.

[0068] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2is C-C alkyl substituted with one or more halo. In some embodiments, R 2 is C1-C6 alkyl substituted with one cyano. In some embodiments, R 2 can be one or more -OR 5 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C1-C6 alkyl substituted with one or more -OH. In some embodiments, R 2 is C1-C6 alkyl substituted with one -OH. In some embodiments, R 2 is one or more -NR 5 R 6 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C1-C6 alkyl substituted with one or more -NH2. In some embodiments, R 2 is C1-C6 alkyl substituted with one -NH2. In some embodiments, R 2 is one -S(O)2R 5 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C1-C6 alkyl substituted with one -S(O)2H. In some embodiments, R 2 is one -S(O)2OR 5 In some embodiments, R is a C1-C6 alkyl substituted with 2 is C-C alkyl substituted with one -S(O)OH. In some embodiments, R 2 is unsubstituted C1-C6 alkyl. In some embodiments, R 2 is -CH3. In some embodiments, R 2 is -CH2CH3. In some embodiments, R 2 is -CH2CH2CH3.

[0069] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6, -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted C2-C6 alkenyl.

[0070] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted C2-C6 alkynyl.

[0071] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted C3-C6 cycloalkyl.

[0072] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is an unsubstituted C3-C6 heterocycloalkyl.

[0073] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R is phenyl optionally substituted with one or more of 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is phenyl substituted with one or more halo. In some embodiments, R 2 can be one or more -OR 5 In some embodiments, R 2 is phenyl substituted with one or more -OH. In some embodiments, R 2 is unsubstituted phenyl.

[0074] In some embodiments, R 2 is halo, cyano, -OR 5 , -NR 5 R 6 , -S(O)2R 5 , or -S(O)2OR 5 In some embodiments, R 2 is unsubstituted -C1-C6 alkyl-phenyl.

[0075] In some embodiments, R 2 -OR 5 In some embodiments, R 2 is -OH. In some embodiments, R 2 -NR 5 R 6 In some embodiments, R 2 is -NH2.

[0076] In some embodiments, R 2is hydrogen.

[0077] In some embodiments of the pharmaceutical compositions described herein, the fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (II) is

[0078] [ka]

[0079] [ka] The structure is selected from:

[0080] Peripherally restricted FAAH inhibitors The pharmaceutical compositions described herein comprise a peripherally restricted FAAH inhibitor. In some embodiments, the peripherally restricted FAAH inhibitor is disclosed in U.S. Patent Application Publication No. 2008 / 0306046, the entire contents of which are incorporated herein by reference.

[0081] In some embodiments, the peripherally restricted FAAH inhibitor has formula (X):

[0082] [ka] or a pharma- ceutically acceptable salt thereof, wherein Ring A is a benzene ring, a cyclopentane ring, a cyclohexane ring, a cycloheptane ring, or a 5- to 7-membered nitrogen-containing heterocycle; L is a single bond, a lower alkylene, a lower alkenylene, -N(R 15 )-C(=O)-, -C(=O)-N(R 15 )-, -(lower alkenylene)-C(=O), -O-, or C(=O); R 15 is H or lower alkyl, X is CH or N; R 8 , R 9 , and R 10are each independently (i) a group selected from the group consisting of H, halo, -CN, CF3, lower alkyl, and -O-lower alkyl; (ii) aryl optionally substituted with 1 to 5 groups independently selected from the group consisting of H, halo, -CN, CF3, lower alkyl, and -O-lower alkyl; (iii) a nitrogen-containing heteroaryl optionally substituted with 1 to 5 groups independently selected from the group consisting of H, halo, -CN, -CF3, lower alkyl, and -O-lower alkyl; (iv) R 16 -(lower alkenylene)-O-, (v)R 16 -N-(lower alkenylene)-(R 15 )-,or (vi)R 17 R 18 NC(=O)- is selected from R 16 teeth, (i) aryl optionally substituted with 1 to 5 groups independently selected from the group consisting of H, halo, -CN, -CF3, lower alkyl, and -O-lower alkyl; (ii) a nitrogen-containing heteroaryl optionally substituted with 1 to 5 groups independently selected from the group consisting of H, halo, -CN, -CF3, lower alkyl, and -O-lower alkyl; or (iii) 3- to 8-membered cycloalkyl and R 17 and R 18 are each independently selected from H, lower alkyl, and 3- to 8-membered cycloalkyl; or R 17 and R 18 may form a 3- to 8-membered nitrogen-containing heterocycle together with the nitrogen atom to which it is bonded, R 11 is selected from H, lower alkyl, and oxo (=O); R 12 , R 13 , and R 14One of is -C(=O)-(O)-(lower alkyl) or -COH, and the rest are H.

[0083] In some embodiments, the peripherally restricted FAAH inhibitor is 5-(((4-(4-((3-fluorobenzyl)oxy)phenoxy)piperidin-1-yl)carbonyl)oxy)nicotinic acid. In some embodiments, the peripherally restricted FAAH inhibitor is 5-(((4-(2-phenylethyl)piperidin-1-yl)carbonyl)oxy)nicotinic acid. In some embodiments, the peripherally restricted FAAH inhibitor is 5-(((4-(4-(2-cyclohexylethoxy)phenoxy)piperidin-1-yl)carbonyl)oxy)nicotinic acid. In some embodiments, the peripherally restricted FAAH inhibitor is 5-(((4-((E)-2-phenylvinyl)piperidin-1-yl)carbonyl)oxy)nicotinic acid. In some embodiments, the peripherally restricted FAAH inhibitor is 5-(((4-(3-(1-(6-methylpyridin-2-yl)piperidin-4-yl)propyl)piperidin-1-yl)carbonyl)oxy)nicotinic acid. In some embodiments, the peripherally restricted FAAH inhibitor is 5-(methoxycarbonyl)pyridin-3-yl 4-(2-phenylethyl)piperazine-1-carboxylate. In some embodiments, the peripherally restricted FAAH inhibitor is ASP-3652. In some embodiments, the peripherally restricted FAAH inhibitor is ASP-3652, which is 5-(((4-(2-phenylethyl)piperidin-1-yl)carbonyl)oxy)nicotinic acid.

[0084] compound The compounds of formula (I'), (I), and (II) described herein are amide prodrugs of TRβ agonists. The amide prodrugs described herein are cleaved by fatty acid amide hydrolase (FAAH) to provide the active TRβ agonist. In some embodiments,

[0085] [ka]

[0086] [ka] or a pharma- ceutically acceptable salt or solvate thereof.

[0087] In some embodiments, a pharma- ceutically acceptable excipient and

[0088] [ka]

[0089] [ka] or a pharma- ceutically acceptable salt or solvate thereof.

[0090] method In some embodiments, a method for treating a CNS disease or disorder in a patient in need of such treatment comprises administering to the patient a pharmaceutical composition as described herein, comprising a fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (I') or (I), or a pharma- ceutically acceptable salt or solvate thereof, a pharma-ceutically acceptable excipient, and a peripherally restricted FAAH inhibitor. In some embodiments, a method for treating a CNS disease or disorder in a patient in need of such treatment comprises administering to the patient a pharmaceutical composition as described herein, comprising a fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (I') or (I), or a pharma-ceutically acceptable salt or solvate thereof, a pharma-ceutically acceptable excipient, and a peripherally restricted FAAH inhibitor ASP-3652.In some embodiments, a method of treating a CNS disease or disorder in a patient in need of such treatment comprises administering to the patient a pharmaceutical composition as described herein comprising a fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (I') or (I), or a pharma- ceutical acceptable salt or solvate thereof, a pharma- ceutical acceptable excipient, and a peripherally restricted FAAH inhibitor, wherein the CNS disease or disorder is selected from the group consisting of acute disseminated encephalomyelitis (ADEM), acute hemorrhagic leukoencephalitis (AHL or AHLE), adult Refsum's disease, infantile Refsum's disease, Alexander's disease, Alzheimer's disease, Barro's concentric sclerosis, Canavan's disease, central pontine myelinolysis (CPM), cerebral palsy, cerebrotendinous yellow fever, and the like. In one embodiment, the method is selected from the group consisting of myelofibrosis, chronic inflammatory demyelinating polyneuropathy (CIDP), Devic's syndrome, diffuse myeloclastic sclerosis, encephalomyelitis, Guillain-Barre syndrome, idiopathic inflammatory demyelinating diseases (HDD), Krabbe disease, Leber's hereditary optic neuropathy, leukodystrophy, Marburg multiple sclerosis, Marchiafava-Bienami disease, metachromatic leukodystrophy (MLD), multifocal motor neuropathy (MMN), multiple sclerosis (MS), paraproteinaceous demyelinating polyneuropathy, Pelizaeus-Merzbach disease (PMD), progressive multifocal leukoencephalopathy (PML), tropical spastic paraparesis (TSP), X-linked adrenoleukodystrophy (X-ALD, ALO, or X-linked ALO), and Zellweger syndrome. In some embodiments, a method of treating a CNS disease or disorder in a patient in need of such treatment comprises administering to the patient a pharmaceutical composition described herein comprising a fatty acid amide hydrolase (FAAH)-cleavable prodrug of formula (I') or (I), or a pharma- ceutically acceptable salt or solvate thereof, a pharma- ceutically acceptable excipient, and a peripherally restricted FAAH inhibitor, wherein the CNS disease or disorder is multiple sclerosis.In some embodiments, a method of treating a CNS disease or disorder in a patient in need of such treatment, comprising administering to the patient a pharmaceutical composition described herein comprising a fatty acid amide hydrolase (FAAH)-cleavable prodrug of Formula (I') or (I), or a pharma- ceutically acceptable salt or solvate thereof, a pharma- ceutically acceptable excipient, and a peripherally restricted FAAH inhibitor, wherein the CNS disease or disorder is X-linked adrenoleukodystrophy.

[0091] In some embodiments, a method for treating a CNS disease or disorder in a patient in need of such treatment comprises administering to the patient a pharmaceutical composition as described herein, comprising a prodrug capable of being cleaved by fatty acid amide hydrolase (FAAH) of formula (II), or a pharma- ceutically acceptable salt or solvate thereof, a pharma-ceutically acceptable excipient, and a peripherally restricted FAAH inhibitor. In some embodiments, a method for treating a CNS disease or disorder in a patient in need of such treatment comprises administering to the patient a pharmaceutical composition as described herein, comprising a prodrug capable of being cleaved by fatty acid amide hydrolase (FAAH) of formula (II), or a pharma-ceutically acceptable salt or solvate thereof, a pharma-ceutically acceptable excipient, and a peripherally restricted FAAH inhibitor ASP-3652.In some embodiments, a method of treating a CNS disease or disorder in a patient in need of such treatment comprises administering to the patient a pharmaceutical composition as described herein comprising a fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (II), or a pharma- ceutical acceptable salt or solvate thereof, a pharma- ceutical acceptable excipient, and a peripherally restricted FAAH inhibitor; the CNS disease or disorder is selected from the group consisting of acute disseminated encephalomyelitis (ADEM), acute hemorrhagic leukoencephalitis (AHL or AHLE), adult Refsum's disease, infantile Refsum's disease, Alexander's disease, Alzheimer's disease, Barro's concentric sclerosis, Canavan's disease, central pontine myelinolysis (CPM), cerebral palsy, cerebrotendinous xanthomatosis, The method is selected from chronic inflammatory demyelinating polyneuropathy (CIDP), Devic's syndrome, diffuse myeloclastic sclerosis, encephalomyelitis, Guillain-Barre syndrome, idiopathic inflammatory demyelinating disease (HDD), Krabbe disease, Leber's hereditary optic neuropathy, leukodystrophy, Marburg multiple sclerosis, Marchiafava-Bienami disease, metachromatic leukodystrophy (MLD), multifocal motor neuropathy (MMN), multiple sclerosis (MS), paraproteinaceous demyelinating polyneuropathy, Pelizaeus-Merzbach disease (PMD), progressive multifocal leukoencephalopathy (PML), tropical spastic paraparesis (TSP), X-linked adrenoleukodystrophy (X-ALD, ALO, or X-linked ALO), and Zellweger syndrome. In some embodiments, a method of treating a CNS disease or disorder in a patient in need of such treatment, comprising administering to the patient a pharmaceutical composition described herein comprising a fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (II), or a pharma- ceutically acceptable salt or solvate thereof, a pharma- ceutically acceptable excipient, and a peripherally restricted FAAH inhibitor, wherein the CNS disease or disorder is multiple sclerosis.In some embodiments, a method of treating a CNS disease or disorder in a patient in need of such treatment, comprising administering to the patient a pharmaceutical composition described herein comprising a fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (II), or a pharma- ceutically acceptable salt or solvate thereof, a pharma- ceutically acceptable excipient, and a peripherally restricted FAAH inhibitor, wherein the CNS disease or disorder is X-linked adrenoleukodystrophy.

[0092] Excipients Optional excipients suitable for use in the pharmaceutical compositions described herein include any excipient commonly used in pharmaceuticals and are selected based on compatibility with the active pharmaceutical agent and the release profile characteristics of the desired dosage form. Excipients include, but are not limited to, binders, fillers, flow aids, disintegrants, lubricants, glidants, polymer carriers, plasticizers, stabilizers, surfactants, etc. Summary summaries of excipients described herein can be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1975; Liberman, H. A. and Lachman, L., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins, 1999), which are incorporated herein by reference in their entireties.

[0093] Binders impart cohesiveness to the solid oral dosage formulation. In powder-filled capsule formulations, binders aid in the formation of a fillable plug in a soft or hard shell capsule, and in tablet formulations, they help ensure that the tablet remains intact after compression and ensure uniformity of the blend prior to the compression or filling step. Materials suitable for use as binders in the solid dosage forms described herein include carboxymethylcellulose, methylcellulose (e.g., Methocel®), hydroxypropyl methylcellulose (e.g., Hypromellose USP Pharmacoat-603, hydroxypropyl methylcellulose acetate stearate (Aqoate HS-LF and HS), hydroxyethyl cellulose, hydroxypropyl cellulose (e.g., Klucel®), ethyl cellulose (e.g., Ethocel®), and microcrystalline cellulose (e.g., Avicel®), microcrystalline dextrose, amylose, magnesium aluminum silicate, polysaccharide acids, bentonite, gelatin, polyvinylpyrrolidone / vinyl acetate copolymers, crospovidone, povidone, starch, pregelatinized starch, tragacanth, dextrin, sugars such as sucrose, Examples of suitable gums include, but are not limited to, glycerin (e.g., Dipac®), glucose, dextrose, molasses, mannitol, sorbitol, xylitol (e.g., Xylitab®), lactose, natural or synthetic gums such as acacia, tragacanth, gum ghatti, isapol shell mucilage, starch, polyvinylpyrrolidone (e.g., Povidone® CL, Kollidon® CL, Polyplasdone® XL-10, and Povidone® K-12), larch arabogalactan, Veegum®, polyethylene glycol, waxes, sodium alginate, and the like.

[0094] Fillers or diluents increase the bulk in pharmaceutical formulations. Such compounds include, for example, lactose, starch, mannitol, sorbitol, dextrose, microcrystalline cellulose such as Avicel®, dibasic calcium phosphate dicalcium phosphate dihydrate, tricalcium phosphate, calcium phosphate, anhydrous lactose, spray-dried lactose, pregelatinized starch, compressible sugars such as Di-Pac® (Amstar), hydroxypropyl methylcellulose, sucrose-based diluents, confectioner's sugar, monobasic calcium sulfate monohydrate, calcium sulfate dihydrate, calcium lactate trihydrate, dextrates, hydrolyzed grain solids, amylose, powdered cellulose, calcium carbonate, glycine, kaolin, sodium chloride, inositol, bentonite, and the like.

[0095] Glidants improve the flow properties of powder mixtures and include, for example, colloidal silicon dioxide such as Cab-o-sil®, tribasic calcium phosphate, talc, corn starch, DL-leucine, sodium lauryl sulfate, magnesium stearate, calcium stearate, sodium stearate, kaolin, and micronized amorphous silicon dioxide (Syloid®).

[0096] Lubricants are compounds that prevent, reduce, or inhibit adhesion or friction of materials.Exemplary lubricants include, for example, stearic acid, calcium hydroxide, talc, hydrocarbons such as mineral oil or hydrous vegetable oil such as hydrous soybean oil (Sterotex®), Lubritab®, Cutina®, higher fatty acids and their alkali metal salts and alkaline earth metal salts such as aluminum, calcium, magnesium, zinc, stearic acid, sodium stearate, magnesium stearate, glycerol, talc, wax, Stearowet®, boric acid, sodium acetate, leucine, polyethylene glycol or methoxypolyethylene glycol, for example Carbowax™, sodium oleate, glyceryl behenate (Compitrol 888R), glyceryl palmitostearate (Precirol®), colloidal silica such as Syloid™, starch such as Carb-O-Sil®, corn starch, silicone oil, surfactants, etc. Hydrophilic lubricants include, for example, sodium stearyl fumarate (currently available under the trade name PRUV®), polyethylene glycol (PEG), magnesium lauryl sulfate, sodium lauryl sulfate (SLS), sodium benzoate, sodium chloride, and the like.

[0097] Disintegrants facilitate the breakup or disintegration of pharmaceutical formulations after administration. Examples of disintegrants include starches, such as natural starches, such as corn starch and potato starch, pregelatinized starches, such as National 1551 and Amijel®, or sodium starch glycolate, such as Promogel® and Explotab®, cellulose, such as wood products, microcrystalline cellulose, such as Avicel®, Avicel® PH101, Avicel® PH102, Avicel® PH105, Elcema® P100, Emcocel®, Vivacel®, Ming Examples include Tia®, and Solka-Floc®, methylcellulose, croscarmellose, or crosslinked cellulose, e.g., crosslinked sodium carboxymethylcellulose (Ac-Di-Sol®), crosslinked carboxymethylcellulose, or crosslinked croscarmellose, crosslinked starches such as sodium starch glycolate, crosslinked polymers such as crospovidone, crosslinked polyvinylpyrrolidone, alginates such as alginic acid or salts of alginic acid such as sodium alginate, clays such as Veegum® HV (magnesium aluminum silicate), gums such as agar, guar, locust bean, karaya, pectin, or tragacanth, sodium starch glycolate, resins such as bentonite, natural sponge, cation exchange resins, citrus pulp, sodium lauryl sulfate in starch formulations, and the like.

[0098] Polymeric carriers include compounds such as polyvinylpyrrolidone, e.g., polyvinylpolyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidone K25, or polyvinylpyrrolidone K30, polyvinylpyrrolidone vinyl acetate (PVPVA64), hydroxypropylmethylcellulose (HPMC), hydroxypropylmethylcellulose acetylsuccinate (HPMC AS), and methyl methacrylate polymers (Eudragit polymers).

[0099] Stabilizers include any antioxidant, for example, compounds such as butylated hydroxytoluene (BHT), sodium ascorbate, and tocopherol, buffers, acids, and the like.

[0100] Surfactants include compounds such as sodium lauryl sulfate, sorbitan monooleate, polyoxyethylene sorbitan monooleate, polysorbates, poloxamers, bile salts, glyceryl monostearate, copolymers of ethylene oxide and propylene oxide, e.g., Pluronic® (BASF), d-α-tocopheryl polyethylene glycol succinate (Vitamin E TPGS), and the like.

[0101] The above excipients are provided as examples only and are not intended to include all possible options. Other suitable excipient types include colorants, granulating agents, preservatives, antifoaming agents, plasticizers, etc. In addition, many excipients may have multiple roles or functions or may be classified into multiple groups. This classification is merely illustrative and is not intended to limit the use of a particular excipient.

[0102] The pharmaceutical formulations of the present disclosure are administered to patients (animals and humans) in need of such treatment in dosages that will provide optimal pharmaceutical efficacy. The dosage required for use in any particular application will vary from patient to patient, depending on the nature of the disease being treated, the age and condition of the patient, any concurrent medications or special diets the patient is following, and other factors in addition to the particular pharmaceutical formulation selected, and the appropriate dosage is ultimately at the discretion of the attending physician.

[0103] While preferred embodiments of the present disclosure have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will occur to those skilled in the art without departing from the invention. It is understood that various alternatives to the embodiments of the invention described herein may be utilized in practicing the disclosure. It is intended that the following claims define the scope of the invention, and that methods and structures within the scope of the claims and their equivalents are covered thereby. EXAMPLES

[0104] The following examples are provided for illustrative purposes and are not intended to limit the scope of the claims provided herein. All reference citations in these examples and throughout this specification are incorporated herein by reference for all legal purposes provided thereby. Starting materials and reagents used in the synthesis of the compounds described herein can be synthesized or obtained from commercial sources, such as, but not limited to, Sigma-Aldrich, Acros Organics, Fluka, and Fischer Scientific. In some embodiments, the compounds provided herein are synthesized as described in U.S. Patent Application No. 2019 / 0210950, which is incorporated herein by reference. In some embodiments, the compounds provided herein are synthesized as described in U.S. Patent Application No. 2021 / 0002208, which is incorporated herein by reference. In some embodiments, the compounds provided herein are synthesized as described in WO 2021 / 108549, which is incorporated herein by reference. In some embodiments, the compounds provided herein are synthesized as described in Examples 1-33 below.

[0105] Example 1: Synthesis of 2-(3,5-dichloro-4-{[4-hydroxy-3-(propan-2-yl)phenyl]methyl}phenoxy)-N-(6-methoxypyridin-3-yl)acetamide (Compound 101)

[0106] [ka]

[0107] Step 1: To a solution of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetic acid (compound 100) (100 mg, 0.3 mmol) in DCM (3 mL) was added DMF (cat). The mixture was cooled to 0° C. and oxalyl chloride (57 mg, 0.45 mmol) was added. The mixture was stirred at room temperature for 30 minutes and then concentrated in vacuo to give 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetyl chloride (110 mg, 95% yield) as a yellow solid.

[0108] Step 2: To a solution of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetyl chloride (110 mg, 0.3 mmol) in DCM (2 mL) was added a mixture of 6-methoxypyridin-3-amine (37 mg, 0.3 mmol) and triethylamine (61 mg, 0.6 mmol) in DCM (3 mL). The mixture was stirred at room temperature for 1 h. Water (15 mL) was added and the resulting mixture was extracted with DCM (3 x 20 mL). The combined organic phase was washed with brine (20 mL), dried over Na2SO4, concentrated in vacuo and purified by preparative HPLC to give 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(6-methoxypyridin-3-yl)acetamide (compound 101) (30 mg, 21% yield) as a white solid. LCMS: M+H=475.2.

[0109] Example 2: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrazin-2-yl)acetamide (Compound 102)

[0110] [ka]

[0111] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrazin-2-yl)acetamide (compound 102) was synthesized according to the method of example 1, using pyrazin-2-amine in step 2. LCMS: M+H=446.1.

[0112] Example 3: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(3,4-dimethylisoxazol-5-yl)acetamide (Compound 103)

[0113] [ka]

[0114] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(3,4-dimethylisoxazol-5-yl)acetamide (compound 103) was synthesized according to the method of example 1, using 3,4-dimethylisoxazol-5-amine in step 2. LCMS: M+H=463.1.

[0115] Example 4: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrazin-3-yl)acetamide (Compound 104)

[0116] [ka]

[0117] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrazin-3-yl)acetamide (compound 104) was synthesized according to the method of example 1, using pyrazin-3-amine in step 2. LCMS: M+H=446.1.

[0118] Example 5: Synthesis of N-cyclohexyl-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetamide (Compound 105)

[0119] [ka]

[0120] N-Cyclohexyl-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetamide (compound 105) was synthesized according to the method of example 1, using cyclohexanamine in step 2. LCMS: MH=448.2.

[0121] Example 6: Synthesis of N-(but-2-yn-1-yl)-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetamide (Compound 106)

[0122] [ka]

[0123] Step 1: A sealed tube (50 mL) was charged with 1-bromobut-2-yne (400 mg, 3.0 mmol) and NH3 (10 mL, 7 M in MeOH). The mixture was stirred at 60 °C overnight. The mixture was concentrated in vacuo to give but-2-yn-1-amine hydrobromide (400 mg, 89% yield) as a yellow solid.

[0124] Step 2: N-(but-2-yn-1-yl)-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetamide (compound 106) was synthesized according to the method of example 1 using but-2-yn-1-amine hydrobromide in step 2. LCMS: MH=418.1.

[0125] Example 7: Synthesis of N-(but-2-yn-1-yl)-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-methylacetamide (Compound 107)

[0126] [ka]

[0127] Step 1: To a mixture of N-methylprop-2-yn-1-amine (2.0 g, 29.0 mmol) in THF (20 mL) was added tert-butyl dicarbonate (18.9 g, 87.0 mmol). The mixture was cooled to 40° C. and stirred for 2.0 hours. The mixture was then concentrated in vacuo to give tert-butyl methyl(prop-2-yn-1-yl)carbamate (4.0 g, 82% yield) as a colorless oil.

[0128] Step 2: To a solution of tert-butyl methyl(prop-2-yn-1-yl)carbamate (1.0 g, 5.9 mmol) in DCM (5 mL) was added n-butyllithium (2.5 M / THF) (2.8 mL, 7.1 mmol) at -70°C. The mixture was stirred for 1 h and iodomethane was added over an additional 1.0 h. Water (50 mL) was added and the resulting mixture was extracted with DCM (3 x 20 mL). The combined organic phase was washed with brine (50 mL), dried over Na2SO4, and concentrated in vacuo to give crude tert-butyl but-2-yn-1-yl(methyl)carbamate (1.0 g, 92% yield) as a colorless oil.

[0129] Step 3: To a solution of tert-butyl but-2-yn-1-yl(methyl)carbamate (1.0 g, 5.5 mmol) in DCM (2 mL) was added TFA (1.3 g, 11.0 mmol) for 2.0 h at 0° C. The resulting mixture was concentrated in vacuo to give N-methylbut-2-yn-1-amine (200.0 mg, 44% yield) as a colorless oil.

[0130] Step 4: N-(but-2-yn-1-yl)-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-methylacetamide (compound 107) was synthesized according to the method of example 1, using N-methylbut-2-yn-1-amine in step 2. LCMS: M+H=434.1.

[0131] Example 8: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N,N',N'-trimethylacetohydrazide (Compound 108)

[0132] [ka]

[0133] Step 1: To a mixture of tert-butyl 1-methylhydrazine-1-carboxylate (1.0 g, 6.8 mmol) in acetonitrile (10 mL) was added formaldehyde (37 wt.% in water) (5.26 mL, 68.4 mmol). The mixture was stirred at room temperature for 2 h. After 2 h, sodium cyanoborohydride (860.0 mg, 13.7 mmol) was added to the solution. The mixture was stirred at room temperature for 2 h, then the mixture was quenched with water (20 mL) and extracted with EtOAc (2 x 10 mL). The organic phase was washed with water (20 mL) and brine (20 mL), dried over Na2SO4, concentrated in vacuo, and purified by silica gel column (DCM to DCM / MeOH=10:1) to give tert-butyl 1,2,2-trimethylhydrazine-1-carboxylate (0.1 g, 8.4% yield) as a colorless oil.

[0134] Step 2: To a solution of tert-butyl 1,2,2-trimethylhydrazine-1-carboxylate (0.1 g, 573.9 μmol) in DCM (2 mL) was added HCl (1M in ether) (5.7 mL, 5.7 mmol). The mixture was stirred at room temperature for 1 h and then concentrated in vacuo to give 1,1,2-trimethylhydrazine dihydrochloride (70 mg, 71.1% yield) as a white solid.

[0135] Step 3: 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N,N',N'-trimethylacetohydrazide (compound 108) was synthesized according to the method of example 1 using 1,1,2-trimethylhydrazine dihydrochloride in step 2. LCMS: M+H=425.0.

[0136] Example 9: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrimidin-5-yl)acetamide (Compound 109)

[0137] [ka]

[0138] A solution of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrimidin-5-yl)acetamide (compound 100) (0.1 g, 271 μmol), pyrimidin-5-amine (25.8 mg, 271 μmol), HATU (124 mg, 325 μmol), and DIPEA (112 μL, 2.5 eq., 677 μmol) in DMF (2 mL) was stirred at room temperature for 5.0 h. Water (10 mL) was added and the resulting mixture was extracted with EtOAc (3×10 mL). The combined organic phase was washed with water (2 x 15 mL), brine (15 mL), dried over Na2SO4, concentrated in vacuo and purified by preparative HPLC to give 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrimidin-5-yl)acetamide (compound 109) (20 mg, 44.8 μmol) as a white solid. LCMS: M+H=446.1.

[0139] Example 10: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrimidin-4-yl)acetamide (Compound 110)

[0140] [ka]

[0141] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrimidin-4-yl)acetamide (compound 110) was synthesized using pyrimidin-4-amine according to the method of example 9. LCMS: M+H=446.1.

[0142] Example 11: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-((1R,2S)-2-fluorocyclopropyl)acetamide (Compound 111)

[0143] [ka]

[0144] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-((1R,2S)-2-fluorocyclopropyl)acetamide (compound 111) was synthesized following the method of example 9 using (1R,2S)-2-fluorocyclopropan-1-amine. LCMS: M+H=426.1.

[0145] Example 12: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(3-fluoropropyl)acetamide (Compound 112)

[0146] [ka]

[0147] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(3-fluoropropyl)acetamide (compound 112) was synthesized using 3-fluoropropan-1-amine according to the method of example 9. LCMS: MH=426.1.

[0148] Example 13: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(6-methoxypyrazin-3-yl)acetamide (Compound 113)

[0149] [ka]

[0150] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(6-methoxypyrazin-3-yl)acetamide (compound 113) was synthesized using 6-methoxypyrazin-3-amine according to the method of example 9. LCMS: M+H=476.2.

[0151] Example 14: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyridin-3-yl)acetamide (Compound 114)

[0152] [ka]

[0153] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyridin-3-yl)acetamide (compound 114) was synthesized using pyridin-3-amine according to the method of example 9. LCMS: M+H=445.1.

[0154] Example 15: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyridin-4-yl)acetamide (Compound 115)

[0155] [ka]

[0156] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyridin-4-yl)acetamide (compound 115) was synthesized using pyridin-4-amine according to the method of example 9. LCMS: M+H=445.2.

[0157] Example 16: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrazin-4-yl)acetamide (Compound 116)

[0158] [ka]

[0159] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(pyrazin-4-yl)acetamide (compound 116) was synthesized using pyrazin-4-amine according to the method of example 9. LCMS: M+H=446.2.

[0160] Example 17: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-1-(pyrrolidin-1-yl)ethan-1-one (Compound 117)

[0161] [ka]

[0162] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-1-(pyrrolidin-1-yl)ethan-1-one (compound 117) was synthesized according to the method of example 9 using pyrrolidine. LCMS: M+H=422.1.

[0163] Example 18: Synthesis of 1-(azetidin-1-yl)-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)ethan-1-one (Compound 118)

[0164] [ka]

[0165] 1-(azetidin-1-yl)-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)ethan-1-one (compound 118) was synthesized using azetidine according to the method of example 9. LCMS: M+H=408.1.

[0166] Example 19: Synthesis of N-(tert-butyl)-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetamide (Compound 119)

[0167] [ka]

[0168] N-(tert-butyl)-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetamide (compound 119) was synthesized using 2-methylpropan-2-amine according to the method of example 9. LCMS: M+H=424.1.

[0169] Example 20: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-isobutyl-N-methylacetamide (Compound 120)

[0170] [ka]

[0171] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-isobutyl-N-methylacetamide (compound 120) was synthesized using N,2-dimethylpropan-1-amine according to the method of Example 9. LCMS: M+H=438.2.

[0172] Example 21: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-isobutylacetamide (Compound 121)

[0173] [ka]

[0174] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-isobutylacetamide (compound 121) was synthesized using 2-methylpropan-1-amine according to the method of example 9. LCMS: MH=422.1.

[0175] Example 22: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-isopropyl-N-methylacetamide (Compound 122)

[0176] [ka]

[0177] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-isopropyl-N-methylacetamide (compound 122) was synthesized using N-methylpropan-2-amine according to the method of example 9. LCMS: MH=422.1.

[0178] Example 23: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(2-hydroxyethyl)-N-methylacetamide (Compound 123)

[0179] [ka]

[0180] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(2-hydroxyethyl)-N-methylacetamide (compound 123) was synthesized using 2-(methylamino)ethan-1-ol according to the method of example 9. LCMS: MH=424.1.

[0181] Example 24: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N,N'-dimethylacetohydrazide (Compound 124)

[0182] [ka]

[0183] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N,N'-dimethylacetohydrazide (compound 124) was synthesized using 1,2-dimethylhydrazine according to the method of example 9. LCMS: M+H=411.1.

[0184] Example 25: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(2-fluoroethyl)-N-methylacetamide (Compound 125)

[0185] [ka]

[0186] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(2-fluoroethyl)-N-methylacetamide (compound 125) was synthesized using 2-fluoro-N-methylethan-1-amine according to the method of example 9. LCMS: M+H=428.1.

[0187] Example 26: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-isopropylacetamide (Compound 126)

[0188] [ka]

[0189] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-isopropylacetamide (compound 126) was synthesized using propan-2-amine according to the method of example 9. LCMS: M+H=410.1.

[0190] Example 27: Synthesis of N-cyclobutyl-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetamide (Compound 127)

[0191] [ka]

[0192] N-Cyclobutyl-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetamide (compound 127) was synthesized using cyclobutanamine according to the method of example 9. LCMS: M+H=422.2.

[0193] Example 28: Synthesis of N-allyl-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-methylacetamide (Compound 128)

[0194] [ka]

[0195] N-Allyl-2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-methylacetamide (compound 128) was synthesized using N-methylprop-2-en-1-amine according to the method of example 9. LCMS: MH=420.1.

[0196] Example 29: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-methyl-N-propylacetamide (Compound 129)

[0197] [ka]

[0198] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-methyl-N-propylacetamide (compound 129) was synthesized using N-methylpropan-1-amine according to the method of example 9. LCMS: MH=422.1.

[0199] Example 30: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-ethyl-N-methylacetamide (Compound 130)

[0200] [ka]

[0201] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-ethyl-N-methylacetamide (compound 130) was synthesized using N-methylethanamine according to the method of example 9. LCMS: M+H=410.2.

[0202] Example 31: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-methyl-N-(2,2,2-trifluoroethyl)acetamide (Compound 131)

[0203] [ka]

[0204] To a solution of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)acetic acid (100 mg, 0.3 mmol) in DMF (3 mL) was added 2,2,2-trifluoro-N-methylethan-1-amine (134 mg, 0.9 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) (77 mg, 0.4 mmol), 1-hydroxybenzotriazole (HOBT) (55 mg, 0.4 mmol), and N,N-diisopropylethylamine (105 mg, 0.8 mmol). The mixture was stirred at room temperature overnight. Water (20 mL) was added. The mixture was extracted with EtOAc (2 x 15 mL). The combined organic phase was washed with brine (20 mL), dried over Na2SO4, concentrated in vacuo, and purified by preparative HPLC to give 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-methyl-N-(2,2,2-trifluoroethyl)acetamide (compound 131) (50 mg, 36% yield) as a white solid. LCMS: MH=462.1.

[0205] Example 32: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(2,2-difluoroethyl)-N-methylacetamide (Compound 132)

[0206] [ka]

[0207] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-N-(2,2-difluoroethyl)-N-methylacetamide (compound 132) was synthesized following the method of Example 31 using 2,2-difluoro-N-methylethan-1-amine hydrochloride. LCMS: M+H=446.1.

[0208] Example 33: Synthesis of 2-(3,5-dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-1-morpholinoethan-1-one (Compound 133)

[0209] [ka]

[0210] 2-(3,5-Dichloro-4-(4-hydroxy-3-isopropylbenzyl)phenoxy)-1-morpholinoethan-1-one (compound 133) was synthesized according to the method of Example 31 using morpholine. LCMS: MH=436.0.

[0211] Example 34: FAAH Substrate Evaluation Purified recombinant human FAAH (rhFAAH) was purchased from Cayman Chemical (Ann Arbor, MI, USA). The total volume per incubation was 400 μL containing final 0.5 ng / μL rhFAAH, 1 μM test compound, 1.25% ethanol or 1 μM PF-3845 (FAAH inhibitor), and 0.1% bovine serum albumin in Tris-EDTA buffer (pH 8.0). The positive control was LL-341001. Incubations were performed at room temperature. At 0, 5, 15, 30, and 60 min, aliquots of 30 μL of reaction mixture were removed and mixed with 300 μL of acetonitrile containing 5 ng / mL terfenadine and 10 ng / mL tolbutamide as internal standards to quench the reaction. The resulting mixture was centrifuged at 4000 rpm at 4° C. for 15 min, and 100 μL of the supernatant was subjected to LC-MS / MS analysis to measure the formation of acid metabolites.

[0212] LC-MS / MS analysis A Waters Acquity Ultra Performance LC system was used for sample analysis. Chromatography was performed on a reversed-phase Kinetex 2.6 μm C18 column, 2.1 × 30 mm, 100 Å. Mobile phase A consisted of 0.1% formic acid in water and mobile phase B consisted of 0.1% formic acid in acetonitrile, run at a flow rate of 0.8 mL / min for 2 min for acid metabolites from positive controls and 0.9 mL / min for 1.5 min for acid metabolites of test compounds. Mass spectrometers (API-5500 and API Q Trap 4000 Applied Biosystems / MDS SCIEX Instruments, Framingham, MA, USA) were operated under ESI positive or negative ion MRM mode.

[0213] Data analysis The formation of acid metabolites was monitored and quantified using a single calibration point of 1 μM. The observed rate constants (ke) for acid metabolite formation were calculated by plotting metabolite concentration against incubation time with the slope as ke and are shown in Table 1.

[0214] [Table 1-1]

[0215] [Table 1-2]

[0216] [Table 1-3]

[0217] [Table 1-4]

[0218] [Table 1-5]

[0219] [Table 1-6]

[0220] [Table 1-7]

[0221] [Table 1-8]

[0222] Example 35: In Vitro Stability Evaluation in Mouse Plasma Male CD-1 mouse plasma is purchased from BioIVT (catalog number MSE00PLK2YNN) and thawed in a 37 °C water bath with the pH adjusted to 7.4 on the day of testing. After a 15 min pre-warming period in the 37 °C water bath, 398 µL of plasma is added with an aliquot of 2 µL stock solution of test compound or positive control (propantheline) in dimethyl sulfoxide (DMSO) to achieve a final concentration of 1 µM with 0.5% DMSO. After thorough mixing, the mixture is returned to the 37 °C water bath for incubation. At 0, 15, 30, 60, and 120 min, aliquots of 30 µL of reaction mixture are removed and mixed with 300 µL of acetonitrile containing 5 ng / mL terfenadine and 10 ng / mL tolbutamide as internal standards to quench the reaction. The resulting mixture is centrifuged at 4000 rpm at 4° C. for 15 min, and 100 μL of the supernatant is removed and mixed with 100 μL of water for liquid chromatography-tandem mass spectrometry (LC-MS / MS).

[0223] LC-MS / MS analysis A Shimadzu LC 30-AD HPLC system is used for sample analysis. Chromatography is performed on a reversed-phase Kinetex 2.6 μm C18 column, 3.0 × 30 mm, 100 Å. Mobile phase A contains 0.1% formic acid in water, mobile phase B contains 0.1% formic acid in acetonitrile, with a run time of 2 min. Mass spectrometers (API-4000 and API Q Trap 4500 Applied Biosystems / MDS SCIEX Instruments, Framingham, MA, USA) are operated under electrospray ionization (ESI) positive or negative ion multiple reaction monitoring (MRM) mode.

[0224] Data analysis The percentage of compound remaining at a particular time point is calculated based on the peak area ratio at time 0 (taken as 100%). The observed rate constant (k obs ) and the gradient is k obs The half-life (t 1 / 2 ) is the formula:t 1 / 2 =0.693 / k obs Obey and demand.

[0225] Example 36: In Vivo Tissue Distribution Study in Male CD-1 Mice Male CD-1 mice (n=6 / group), 7-10 weeks of age, are acclimated to the laboratory for a minimum of 3 days prior to dosing in the study. Test compounds are formulated in 1% N-methyl-2-pyrrolidone (NMP) and 1% solutol in phosphate buffered saline (PBS) at 0.1 mg / mL in a clear solution with a dose volume of 10 mL / kg. The peripherally restricted FAAH inhibitor LL-650021 is formulated in 0.5% carboxymethylcellulose in water at 0.1 mg / mL with a dose volume of 10 mL / kg. The concentration of the formulation is determined to meet the acceptance criteria within 20% of the target value.

[0226] Test compounds are administered to non-fasted mice at 1 mg / kg by subcutaneous (SC) injection or oral gavage (PO) with or without pretreatment with 1 mg / kg LL-650021 1 hour prior to test compound administration. Animals (n=2 per time point) are sacrificed by CO2 inhalation 1, 4, and 8 hours after administration. Blood samples (0.3 mL) are collected from the saphenous vein or other suitable site into pre-chilled K2EDTA tubes and placed on moist ice, and brains and livers are harvested. Blood samples are centrifuged at 3200 g for 10 minutes at 4°C, plasma samples are transferred to polypropylene tubes, flash frozen on dry ice, and maintained at or below -60°C until analysis. The tissue is washed with cold saline, blotted dry, weighed, and then homogenized in 15 mM PBS (pH 7.4):methanol (2:1) at a ratio of 1:10 (1 g tissue with 10 mL buffer gives an 11-fold dilution). The tissue homogenate is kept at -60°C or below until analysis.

[0227] Sample extraction Plasma and tissue homogenates are extracted by protein precipitation. Aliquots of 10-50 µL of plasma or 40-50 µL of tissue homogenates are subjected to protein precipitation by adding 200-800 µL of acetonitrile containing internal standards (10 ng / mL of LL-120001, and 100 ng / mL of celecoxib, dexamethasone, glyburide, labetalol, tolbutamide, and verapamil), vortex-mixing at 800 rpm for 10 min, and centrifuging at 4000 rpm and 4 °C for 15 min. The supernatants are transferred to a 96-well plate and centrifuged at 4000 rpm and 4 °C for 5 min before injection for LC-MS / MS analysis, or 200 µL of the supernatant is transferred to a 96-well plate, evaporated to dryness under a stream of nitrogen at 25 °C, and reconstituted in 50 µL of 70% acetonitrile. Vortex mix at 800 rpm for 10 min and centrifuge at 4000 rpm at 4° C. for 5 min before injection for LC-MS / MS analysis.

[0228] LC-MS / MS analysis An Acquity Ultra Performance LC system from Waters is used for sample analysis. Separation is performed on an ACQUITY UPLC BEH C18 column (50 × 2.10 mm, 1.7 μm) at 50 °C with a flow rate of 0.6 mL / min. Mobile phase A consists of 2 mM ammonium acetate in 5:95 methanol:water, and mobile phase B consists of 2 mM ammonium acetate in 95:5 acetonitrile:water. The chromatography uses a linear gradient starting at 2% mobile phase B, going from 2% to 90% mobile phase B over 2.6 min, with a 0.2 min hold during the 90% B wash, and a 0.2 min re-equilibration at 2% B. Aliquots of 2-9 μL of sample are injected. The mass spectrometer (API-6500, Applied Biosystems / MDS SCIEX Instruments, Framingham, MA, USA) is operated under ESI in positive or negative ion MRM mode.

[0229] Example 37: In vitro selectivity of prodrugs and agonists to TRβ receptors The potency and selectivity of LL-341070, a thyromimetic prodrug of formula (I') described herein that delivers LL-341070A, a potent and selective small molecule agonist of thyroid hormone receptor (TR) β after fatty acid amide hydrolase (FAAH)-mediated conversion, against the thyroid hormone β receptor (TRβ) was evaluated. In vitro potency was determined by dosing test compounds in a luciferase-based TR reporter cell line using thyroid hormone (T3) as a positive control. Table 2 shows the potency profiles of LL-341070 prodrugs and LL-341070A active metabolites against TRβ and TRα, measured at half maximal effective concentrations (EC50), with selectivity measurements adjusted for the TRα bias of T3 in the assay. Both LL-341070 and LL-341070A exhibit enhanced selectivity for TRβ, and LL-341070A exhibits enhanced potency.

[0230] [Table 2]

[0231] Example 38: LL-341070A Enhances Differentiation of Oligodendrocyte Progenitor Cells In Vitro To profile LL-341070A, in vitro oligodendrocyte precursor cell (OPC) assays were performed on primary OPC cultures generated from E14.5 PLP-EGFP C57B1 / 6 mouse embryo brains. Thyroid hormone (T3), known to induce OPC differentiation and remyelination, was used as a positive control at 10 ng / mL. Primary OPC cultures were treated with LL-341070A compounds at concentrations ranging from about 1 mM to about 10 mM. After inducing OPC differentiation with or without LL-341070A compounds for 5 days (N=6 / concentration), cells were fixed and stained for myelin basic protein (MBP) and normalized to total cell number.

[0232] Figure 1 shows the active metabolite of LL-341070A, a prodrug of LL-341070, in enhancing oligodendrocyte differentiation in an oligodendrocyte precursor cell assay in vitro (EC50=1.4 nM). Enhanced oligodendrocyte differentiation was shown to be relatively consistent as a function of LL-341070A treatment concentration.

[0233] Example 39: Thyromimetic Treatment Enhances 24-OHC Synthesis In Vivo The ability of thyromimetic drugs to accelerate the remyelination process in vivo was evaluated by measuring the fractional synthesis of 24-hydroxycholesterol (24-OHC) in rat brain after cuprizone-induced demyelination. As shown in Figure 2, n=10F rats / group were evaluated in the cuprizone demyelination model. The effect of thyromimetic treatment on the fractional synthesis of 24OHC was measured for brain and plasma using deuterated water labeling of 24OHC for the cuprizone demyelination model after withdrawal from a 0.6% cuprizone diet during the period of active remyelination. Upon withdrawal from a 0.6% cuprizone diet for 3 weeks, rats were given 30 or 100 μg / kg LL-341070 along with deuterated water, and then 24-OHC deuterium enrichment and labeling patterns in the cortex and corpus callosum were measured. LL-341070 induced a dose-dependent increase in deuterated 24-OHC compared to vehicle controls, thus indicating an increase in myelin synthesis rate. As shown in Figure 2, thyroid stimulation treatment enhances the fractional synthesis rate (FSR) of 24S-hydroxycholesterol in the brain and demonstrates a strong correlation with the 24OHC FSR in plasma. 24OHC fractional synthesis rates (FSR) were calculated based on data collected from tissues and plasma that underwent alkaline hydrolysis and derivatization for GC / MS or LC / MS analysis of deuterated 24OHC. Plasma analysis of 24OHC was measured by LC / MS as fraction labeled relative to total plasma 24OHC by Ardena Biosciences. Fractional synthesis was calculated by mass isotope distribution analysis using precursor 2H enrichment in body water from liver palmitate 6. Statistics: Data were analyzed by one-way ANOVA with Tukey's multiple comparison test and are expressed as mean / -SEM. *p<0.05.

[0234] Having confirmed the compound's potency, pharmacokinetics, and target engagement with LL-341070, we then tested its efficacy in remyelination models including oligodendrocyte progenitor cell differentiation in vitro and experimental autoimmune encephalitis in vivo.

[0235] Example 40: Engagement of TRβ in the Brain Increases Expression of T3 Target Genes In Vivo Figure 3 shows that TR-b target engagement in the brain is demonstrated by increased expression of T3-responsive target genes in vivo. Single PO administration of LL-341070 (ranging from 0.1 μg / kg to 300 μg / kg) or T3 (approximately 300 μg / kg) to male C57BL / 6 mice increases expression of Hr, Dio3, Klf9 (quantified by QuaniPlex) and the combined mean log2 fold change in the brain. Klf9, a T3-responsive gene associated with myelin regeneration in vitro, is upregulated at various treatment concentrations. This increased expression was confirmed in the brain of the rat cuprizone model (discussed previously) following 21-day repeated administration of LL-341070 at 30 μg / kg or 100 μg / kg, or 21-day repeated administration of T3 administered at 300 μg / kg (quantified by Nanostring). Interestingly, the expression of Dio3 was enhanced by repeated administration.

[0236] Example 41: In Vivo Tissue Distribution Demonstrates Enhanced Brain Exposure of Active Compound Compared to Prodrug The in vivo brain exposure of the active compound compared to the prodrug was evaluated by tissue distribution (TD) assays on mouse and rat cuprizone models and measured as brain-to-plasma brain exposure ratios after thyroid stimulation treatment. As shown in Table 3, a single PO dose of LL-341070 (100 μg / kg) or LL-341070A (100 μg / kg) demonstrated enhanced brain exposure of the active compound LL-341070A compared to the prodrug LL-341070, measured in the brain and plasma of male C57BL / 6 mice, resulting in a brain-to-plasma AUC ratio of greater than 1 for LL-341070A, where AUC is from 0 to 24 hours. The data show that the AUC of LL-341070A in the brain is approximately 7-fold higher than the prodrug LL-341070. Table 3 also shows the brain-to-plasma AUC ratios. As shown in FIG. 4, repeated dosing of LL-341070 (30 μg / kg or 100 μg / kg) or LL-341070A (30 μg / kg or 100 μg / kg) for 21 days demonstrates enhanced brain exposure of the active compound LL-341070A compared to the prodrug LL-341070, as measured in the brain and plasma in a rat cuprizone model 4 hours after the final dose.

[0237] [Table 3]

[0238] Example 42: LL-341070 Improves In Vivo Clinical Scoring and Histology in a Murine EAE Model As shown in Figure 5, the efficacy of LL-341070 was evaluated against a murine prophylactic experimental EAE model in which disease onset occurs 8–18 days after induction of autoimmune encephalitis (EAE) with MOG35-55 / CFA+PTX and EAE is scored 7–28 days after induction. In the EAE model, n=12F C57B1 / 6 mice / group were evaluated that received LL-341070 (10 μg / kg–100 μg / kg) or vehicle PO daily after EAE induction. LL-341070 administered daily in a prophylactic paradigm dose-dependently improved the median day of disease onset and reduced maximum disease severity. Histological analysis of spinal cords 28 days after immunization showed a reduction in inflammatory foci, apoptotic cell numbers, and demyelinated area by H&E and MBP staining. LL-341070 ameliorates mean clinical scoring and histological endpoints of inflammation and demyelination in a mouse EAE model. Clinical scores were adjudicated by a blinded observer. Histology was performed on spinal cord samples (demyelination score assessed by % demyelinated area in anti-MBP staining, inflammatory foci means number of groups of >20 cells per section in H&E staining). Statistics: Median days of EAE onset compared using Wilcoxon survival test.

[0239] Example 43: FAAH expression is enriched in the brain As shown in FIG. 6, brain-specific thyromimetic prodrugs activated by fatty acid amide hydrolysis (FAAH) (such as ABX-002, which is activated to ABX-002A, compound 1 described herein) were utilized to elucidate the mechanism by which thyromimetics disrupt the thyroid hormone axis (THA). Altering the delivery of potent thyromimetics helped identify whether feedback control on THA originates from central (hypothalamic) or peripheral (pituitary) mechanisms, potentially enhancing the therapeutic index of thyromimetics. These studies were performed using recombinant FAAH, tissue-derived S9 fractions, in vivo tissue distribution (TD), gene expression in brain and liver, and effects on T4 in mice as a marker of THA disruption. Northern blot assays confirmed that FAAH was expressed across species (rodent and human) and that relative mRNA FAAH expression was enhanced in the brain. FAAH specific activity (AMC assay cleavage) from tissue-derived S9 fractions of various organs (liver, brain, small intestine) across species (mouse, rat, non-human primate, human), calculated as a percentage of liver activity, was found to be increased in human and non-human primate brain.

[0240] Example 44: FAAH Expression Enhances ABX-002A Delivery to the Brain To assess delivery, concentrations of ABX-002A in brain, liver, kidney, lung, and heart were measured 1 hour after SC administration of 30 different prodrugs of ABX-002A. As shown in Figure 7, the brain-to-plasma ratios were increased compared to ABX-002A with the prodrug, but the tissue-to-plasma ratios in peripheral organs (liver, kidney, lung, and heart) showed a linear (constant) relationship between tissue and plasma. The data show that FAAH is highly expressed in the CNS and that the ABX prodrugs enhanced active metabolite delivery to the brain by more than 30-fold, with brain-to-plasma ratios exceeding 1. In organs other than the brain, the data show that tissue concentrations were driven by the plasma concentration of the active metabolite ABX-002A.

[0241] Example 45: Global and peripheral FAATT inhibitors alter metabolite distribution in mice The ability of globally-penetrant and peripherally-restricted FAAH inhibitors (GFI and PFI, respectively) to alter the distribution of ABX-002 and ABX-002A was evaluated. Table 4 shows the potency profiles (apparent IC) of peripherally and global FAAH inhibitors LL-650177 (PFI), URB9373 (PFI), and PF-044578454 (GFI) obtained after 30 min preincubation with recombinant human FAAH and 7-amino-4-methylcoumarin (AMC). 50 s (measured in nM) are shown.

[0242] [Table 4]

[0243] Figure 8 shows the concentrations of the prodrug (ABX-002) in plasma, liver, and brain when co-administered with or without PFI or GFI. Prodrug levels were unchanged or only slightly increased with FAAH inhibition. Active metabolite (ABX-002A) levels were reduced in plasma and liver with PFI, and in all organs with GFI. Table 5 shows the inhibition of active metabolite (LL-650177 or PF-044578454) with AUC in plasma, liver, and brain after co-administration of the prodrug (ABX-002). Tissue distribution studies on mice confirm global and peripheral inhibition of FAAH.

[0244] [Table 5]

[0245] Example 46: Induction of T3-regulated genes considering prodrugs and FAAH inhibitors Female C57BL / 6 mice (n=5 / group), 6-8 weeks of age, were acclimated to the laboratory for at least 3 days prior to dosing in this study. Non-fasted mice were given a single dose of PFI or vehicle orally (PO) on day 0 at time=1 hr. A single dose of 5mL / kg was administered based on the most recent body weight taken once during the study. Following PFI or vehicle dosing, animals received a single dose of test article at time=0 hr. One group (n=5) received 300ug / kg T3 PO only at time=0 hr. Approximately 4 hours after test article administration (t=4 hr), animals were humanely sacrificed and brain, liver, heart, pituitary, spinal cord, and plasma samples were collected.

[0246] Sample processing a. Expression Analysis Samples - Multiple organs were harvested at endpoint and tissues were immediately processed as described below.

[0247] i. Brain: The skull of each mouse was opened and the brain was removed. The cerebellum was dissected, the cerebral cortex was cut in half sagittally, and the left half was collected. After rinsing extraneous blood from the tissue with ice-cold 0.9% NaCl, the cerebral cortex specimen was placed in a tube containing 1.2 mL of pre-chilled RNALater and stored at 4°C.

[0248] ii. Liver: One liver biopsy (100-150 mg) was taken from each mouse from the left hepatic lobe. After rinsing extraneous blood from this biopsy with ice-cold 0.9% NaCl, the sample was placed in 1.2 mL pre-chilled RNALater and stored at 4°C. iii. Left Ventricle: Left ventricular (LV) blood from each mouse was removed using the PBI standard method and half of the LV free wall was collected. After rinsing extraneous blood from the tissue with ice-cold 0.9% NaCl, the LV free wall was placed in 1.2 mL pre-chilled RNALater and stored at 4°C. LV tissue was kept in PBI for possible future analysis or until the appropriate genes could be identified by 6 months after the end of the in-life phase of the study. The nature of the sample was confirmed before disposal.

[0249] iv. Pituitary: After removal of the brain from each mouse, the pituitary was harvested. After rinsing the pituitary of any extraneous blood with ice-cold 0.9% NaCl, the specimen was placed in 0.15 mL of pre-chilled RNALater and stored at 4°C. Pituitary tissue was kept in PBI for possible future analysis or until appropriate gene identification could be achieved up to 6 months after the end of the in-life phase of the study. The nature of the samples was confirmed before disposal.

[0250] b. Pharmacokinetic Samples - At endpoint, blood and tissue specimens were processed immediately as described below. Samples for PK analysis were kept in PBI at -80°C for up to 90 days after the conclusion of the in-life phase of the study.

[0251] i. Plasma: Whole blood (approximately 300 μL) was collected on K3EDTA by cardiac puncture under isoflurane anesthesia. Blood was immediately placed on moist ice. After completion of the collection procedure, blood was centrifuged at 10,000×g for 10 min at 4° C. Plasma (approximately 125 μL) was aliquoted into appropriately labeled tubes and flash frozen.

[0252] ii. Liver: For each mouse, one liver biopsy (30-50 mg) was taken from the left hepatic lobe. After rinsing extraneous blood from the biopsy with ice-cold 0.9% NaCl, the sample was placed in an appropriately labeled tube and flash frozen in liquid nitrogen.

[0253] iii. Brain: For each mouse, a midbrain biopsy (30-50 mg) was taken from the right cerebral cortex. After rinsing extraneous blood from the tissue with ice-cold 0.9% NaCl, the biopsy was placed in an appropriately labeled tube and flash frozen.

[0254] iv. Left ventricle: The left ventricle (LV) of each mouse was cleared of blood using PBI standard techniques and half of the LV free wall was collected. After rinsing extraneous blood from the tissue with ice-cold 0.9% NaCl, the LV free wall was placed in an appropriately labeled tube and flash frozen.

[0255] Target Engagement Changes in expression of selected genes identified by transcriptome analysis were measured from purified RNA using a hybridization-based in situ RNA quantification method (NanoString, Seattle, WA). Briefly, fresh tissues were collected in RNALater™ stabilization solution, catalog number AM7021 (ThermoFisher Scientific, Carlsbad, CA) and frozen at -20°C until subjected to RNA extraction. Whole blood was collected by terminal cardiac puncture into MiniCollect K2EDTA tubes, catalog number 450480 Greinder Bio-one GmbH (Kremsmunster, Australia) and processed to plasma by centrifugation at 2000xg for 10 minutes at 4°C. For RNA extraction, tissues were homogenized using TRIzol Reagent, catalog number 15596026 (ThermoFisher Scientific) using a bead homogenizer, RNA was extracted according to the manufacturer's protocol, and purified using Econospin RNA Mini Spin Columns For RNA (Ephoch Life Sciences, Missouri City, TX, catalog number 1940-250) according to the manufacturer's protocol. Specific gene probes were designed by the NanoString Bioinformatics Team using target sequences identified based on the NCBI Reference Sequence (RefSeq) database. Custom probes were synthesized by Integrated DNA Technologies (IDT, Coralville, IA). mRNA expression was analyzed on the nCounter® SPRINT Profiler NanoString system using a multiplexing approach with the nCounter PlexSet-12 Reagent Pack, catalog number PS-GX-PTK-12 (CSO), according to the manufacturer's protocol (NanoString, Inc, Seattle, WA).

[0256] Data analysis T3 target genes are increased after a single dose of drug, and relative activity in brain vs. liver is determined by prodrug and / or FAAH inhibition. Relative activity in brain vs. liver (as a marker of peripheral activity) shifts by more than 1500-fold across different dosing paradigms. Figures 9A, 9B, and 9C show induction of T3-regulated genes in brain (blue) and liver (orange) 4 hours after a single dose of (A) active metabolite, (B) prodrug alone, or (C) prodrug + PFI (URB937). RNA was analyzed by Nanostring as follows: mean fold changes of multiple genes were calculated on a log2 scale and normalized to data obtained with 300 mg / kg T3. PFI administration reduced the potency of the prodrug on activation of T3-regulated genes in liver by more than 10-fold, without affecting activity or exposure in the brain. PFI also reduced potency against THA, consistent with negative feedback based on circulating peripheral metabolites rather than brain exposure. Thus, the use of PFI allowed separation of on-target brain effects from effects on the THA.

[0257] Example 47: T4 Parallels Peripheral Activity Female C57BL / 6 mice (n=5 / group), 6-8 weeks of age, were acclimated to the laboratory for at least 3 days prior to dosing in the study. Mice were dosed at 5 mL / kg based on most recent body weight taken once during the study period. Mice were enrolled in weight-adjusted treatment cohorts based on most recent body weight taken once during the study period. Mice were given a single oral (PO) dose of PFI or vehicle at time=1 h daily for 7 days (n=5 / group). Following administration of PFI (100 μg / kg) or vehicle (10 mL / kg, PO), animals were given the test article daily at time=0 h. Test article was administered at one of eight dose levels (0.1, 0.3, 1, 3, 10, 30, 100, or 300 μg / kg) for a total of seven doses. Mice were dosed PO, QD with (A) active metabolite, (B) prodrug alone, (C) prodrug + PFI (LL-650177), or (D) prodrug + GFI for 7 days. Approximately 4 or 8 hours after test article administration (t=4 hours or t=8 hours), animals were humanely sacrificed using standard procedures and brain, liver, and plasma samples were collected. RNA from samples taken 4 hours after the final dose was quantified using a hybridization-based in situ RNA quantification method (NanoString, Seattle, WA) as described below. RNA from samples taken 8 hours after the final dose was quantified using a hybridization-based in situ RNA quantification method (QuantiGene Plex) as described below. On the final day of dosing, mice were dosed on a timetable to mitigate the impact of circadian effects on thyroid hormone-sensitive gene expression. For this reason, the "time of day" was balanced at the time of final sacrifice for treatment groups. Mice were anesthetized 4 or 8 hours after the last dose, bled by retroorbital puncture, and sacrificed using standard procedures. Tissues were harvested immediately after sacrifice and processed according to the following procedures.

[0258] Sample processing a. Expression Analysis Samples - Multiple organs were harvested at endpoint and tissues were immediately processed as described below.

[0259] i. Brain: The skull of each mouse was opened and the brain was removed. The cerebellum was dissected, the cerebral cortex was cut in half sagittally, and the left half was collected. After rinsing extraneous blood from the tissue with ice-cold 0.9% NaCl, the cerebral cortex specimen was placed in a tube containing 1.2 mL of pre-chilled RNALater and stored at 4°C.

[0260] ii. Liver: One liver biopsy (100-150 mg) was taken from each mouse from the left hepatic lobe. After rinsing extraneous blood from this biopsy with ice-cold 0.9% NaCl, the sample was placed in 1.2 mL pre-chilled RNALater and stored at 4°C. iii. Left Ventricle: Left ventricular (LV) blood from each mouse was removed using the PBI standard method and half of the LV free wall was collected. After rinsing extraneous blood from the tissue with ice-cold 0.9% NaCl, the LV free wall was placed in 1.2 mL pre-chilled RNALater and stored at 4°C. LV tissue was kept in PBI for possible future analysis or until the appropriate genes could be identified by 6 months after the end of the in-life phase of the study. The nature of the sample was confirmed before disposal.

[0261] iv. Pituitary: After removal of the brain from each mouse, the pituitary was harvested. After rinsing the pituitary of any extraneous blood with ice-cold 0.9% NaCl, the specimen was placed in 0.15 mL of pre-chilled RNALater and stored at 4°C. Pituitary tissue was kept in PBI for possible future analysis or until appropriate gene identification could be achieved up to 6 months after the end of the in-life phase of the study. The nature of the samples was confirmed before disposal.

[0262] b. Pharmacokinetic Samples - At endpoint, blood and tissue specimens were processed immediately as described below. Samples for PK analysis were kept in PBI at -80°C for up to 90 days after the conclusion of the in-life phase of the study.

[0263] i. Plasma: Whole blood (approximately 300 μL) was collected on K3EDTA by cardiac puncture under isoflurane anesthesia. Blood was immediately placed on moist ice. After completion of the collection procedure, blood was centrifuged at 10,000×g for 10 min at 4° C. Plasma (approximately 125 μL) was aliquoted into appropriately labeled tubes and flash frozen.

[0264] ii. Liver: For each mouse, one liver biopsy (30-50 mg) was taken from the left hepatic lobe. After rinsing extraneous blood from the biopsy with ice-cold 0.9% NaCl, the sample was placed in an appropriately labeled tube and flash frozen in liquid nitrogen.

[0265] iii. Brain: For each mouse, a midbrain biopsy (30-50 mg) was taken from the right cerebral cortex. After rinsing extraneous blood from the tissue with ice-cold 0.9% NaCl, the biopsy was placed in an appropriately labeled tube and flash frozen.

[0266] iv. Left ventricle: The left ventricle (LV) of each mouse was cleared of blood using PBI standard techniques and half of the LV free wall was collected. After rinsing extraneous blood from the tissue with ice-cold 0.9% NaCl, the LV free wall was placed in an appropriately labeled tube and flash frozen.

[0267] Target Engagement Tissue samples were prepared for biochemical analysis by cryopowdering on liquid nitrogen and lysed using standard methods from PBI. Expression changes (mRNA expression) of selected genes identified by transcriptome analysis were measured from purified RNA using hybridization-based in situ RNA quantification methods (NanoString or QuantiGene Plex). Expression data of target genes were expressed as ratios to the geometric mean of appropriately expressed normalized genes. Briefly, fresh tissues were collected in RNALater™ stabilization solution, catalog number AM7021 (ThermoFisher Scientific, Carlsbad, CA) and frozen at -20°C until subjected to RNA extraction. Whole blood was collected by terminal cardiac puncture into MiniCollect K2EDTA tubes, catalog number 450480 Greinder Bio-one GmbH (Kremsmunster, Australia) and processed to plasma by centrifugation at 2000xg for 10 min at 4°C. For RNA extraction, tissues were homogenized in TRIzol Reagent, catalog number 15596026 (ThermoFisher Scientific) using a bead homogenizer, RNA was extracted according to the manufacturer's protocol, and purified using Econospin RNA Mini Spin / Columns For RNA (Ephoch Life Sciences, Missouri City, TX, catalog number 1940-250) according to the manufacturer's protocol. Specific gene probes were designed by the NanoString Bioinformatics Team using target sequences identified based on the NCBI reference sequence (RefSeq) database. Custom probes were synthesized by Integrated DNA Technologies (IDT, Coralville, IA). mRNA expression was analyzed on the nCounter® SPRINT Profiler NanoString system using a multiplexing approach with the nCounter PlexSet-12 Reagent Pack, catalog number PS-GX-PTK-12 (CSO), according to the manufacturer's protocol (NanoString, Inc, Seattle, WA).

[0268] T4 analysis T4 was measured in the final plasma samples using an ELISA kit (Biovision, Inc., Thyroxine [T4] [Mouse / Rat] ELISA Kit, Catalog Number: K7421-100). The assay was performed according to the manufacturer's instructions with minor modifications based on previous assay validation efforts. Briefly, a seven-point standard curve of T4, provided at dilutions in assay buffer (25, 15, 10, 5, 2, 1 μg / dL), was prepared in duplicate for each assay. Plasma samples (undiluted), blanks (assay buffer), and standards were added to separate wells in a 96-well plate pre-coated with T4 capture antibody, followed by the addition of T4 enzyme conjugate to each well. The plate was then mixed by gentle shaking (600 rpm) for 20-30 seconds, then covered with acetate plate seals and incubated at room temperature (RT) with gentle shaking (600 rpm) for 1 hour. The plate contents were aspirated and washed three times with 1X wash buffer, then blotted with paper towels to remove excess liquid. TMB substrate was then added to each well, the plate was secured with an acetate seal, and incubated for 15 minutes at room temperature, protected from light. Stop solution was then added to each well, and the plate was gently shaken to mix the solutions. Absorbance was read at 450 nm within 15 minutes of adding stop solution using a Varioskan Lux plate reader (ThermoFisher Scientific, Carlsbad, CA). Relative optical density (OD) was background corrected against blank samples and the standard curve. T4 concentrations were interpolated using a four-parameter curve-fitting method. Unknown sample concentrations were determined using GraphPad Prism software (GraphPad Prism 9.0.2, GraphPad Software, San Diego, CA).

[0269] Data analysis Figures 10A, 10B, 10C, and 10D show gene expression in the brain (blue) and liver (orange) and the effect on T4 (gray) 4 or 8 hours after the last dose in mice treated with (A) active metabolite, (B) prodrug alone, (C) prodrug + PFI (LL-650177), or (D) prodrug + GFI PO, QD for 7 days. Both the prodrug and active metabolite reduce T4 levels 7 days after treatment. Table 6 shows the ED by treatment type. 50 Values ​​are reported in μg / kg.

[0270] [Table 6]

[0271] T4 was used as a marker of effect on THA. T4 parallels peripheral activity rather than CNS activation of target genes. Negative regulation of T4 by thyromimetics does not appear to be primarily centrally mediated, as effects on THA and hepatic gene expression more closely parallel plasma distribution rather than exposure or activity in the CNS, suggesting a primarily pituitary-driven effect. Combination of thyromimetics prodrugs with PFI may further enhance thyromimetics delivery to the brain and maximize centrally targeted distribution.

[0272] Example 48: Peripheral exposure to ABX-002A predicts efficacy in THA The effect of THA in relation to plasma ABX-002A was examined in both mice and NHPs. Mice: Using the data from female C57BL / 6 mice from Example 14 above, exposure to amide and acid was calculated based on PK data from independent experiments such as those detailed in Example 12. PK was performed on a single dose only, with other doses calculated proportionally. Non-human primates (NHPs): Plasma pharmacokinetics and effects on the thyroid hormone axis were measured in non-naive cynomolgus monkeys (n=3 / group) administered ABX-002 at 10, 30, 100, or 300 μg / kg daily for 7 days. ABX-002 was formulated in 0.1% NMP / 0.1% Solutol and administered 5 mL / kg PO. Blood samples were collected at 0, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours on days 1 and 7 and analyzed by LCMS for ABX-002, LL-340001, T3, and T4 levels (as described above). TSH was measured by immunoassay.

[0273] Bioassays for T3 and T4 in NHP serum were performed with surrogate matrix, QC, double blanks, and blanks. Standard curve samples were prepared by adding 5 μL WS to 50 μL blank surrogate serum. QC samples were prepared by adding 5 μL WS to 50 μL blank surrogate serum. Unknown samples were added with 5 μL DMSO. All calibrator standard, QC, sample, and blank wells received 500 μL of IS working solution in methanol (2.5 ng / mL T3-13C6 and 25 ng / mL T4-13C6), while 500 μL blank methanol was added to all double blanks. After transferring 400 μL of supernatant, samples were evaporated under N2 gas and reconstituted with 100 μL of 80% aqueous methanol.

[0274] Figures 11A, 11B, and 11C show T4 inhibition as a function of (A) dose, (B) plasma prodrug AUC, or (C) plasma active metabolite AUC at 7 days post-treatment in mice (orange) or non-human primate NHPs (blue). T4 levels at day 7 were normalized to the day 1 levels for each animal relative to exposure in the same animal. Overall, the curves depicted in Figure 11C show that the relationship between efficacy of THA and plasma ABX-002A exists in both mice and NHPs, with peripheral exposure to the active metabolite being a better predictor of efficacy to THA than either dose or plasma prodrug exposure.

Claims

1. Formula (I'): 【Chemistry 1】 A fatty acid amide hydrolase (FAAH) cleavable prodrug of the formula: R １ and R ２ each independently represents hydrogen, -OR ５ , -NR ５ R ６ , C １ -C ６ Alkyl, C ２ -C ６ Alkenyl, C ２ -C ６ Alkynyl, C ３ -C ６ Cycloalkyl, C ３ -C ６ Heterocycloalkyl, phenyl, and -C １ -C ６ alkyl-phenyl, １ -C ６ Alkyl, the C ２ -C ６ Alkenyl, the above C ２ -C ６ Alkynyl, the above C ３ -C ６ Cycloalkyl, the C ３ -C ６ heterocycloalkyl, the phenyl, and the -C １ -C ６ Alkyl-phenyl is halo, cyano, -OR ５ , -NR ５ R ６ , -S(O) ２ R ５ or -S(O) ２ OR ５ optionally replaced by one or more of R ３ and R ４ is independently selected from: -F, -Cl, -Br, and -I; R ５ and R ６ are independently hydrogen and C １ -C ６ alkyl, R ７ and R ８ is independently selected from hydrogen, -F, -Cl, -Br, and -I, and a pharma- ceutically acceptable excipient, or a pharma- ceutically acceptable salt or solvate thereof; The pharmaceutical composition, or a pharma- ceutically acceptable salt or solvate thereof, further comprising a peripherally restricted FAAH inhibitor.

2. The R ７ The pharmaceutical composition of claim 1 , or a pharma- ceutically acceptable salt or solvate thereof, wherein R is hydrogen and said R 8 is hydrogen.

3. The pharmaceutical composition of claim 1, or a pharma- ceutically acceptable salt or solvate thereof, wherein R 1 is hydrogen.

4. The pharmaceutical composition of claim 3, or a pharma- ceutically acceptable salt or solvate thereof, wherein R2 is C1-C6 alkyl substituted with one or more of halo, cyano, -OR5, -NR5R6, -S(O)2R5, or -S(O)2OR5.

5. The pharmaceutical composition according to claim 4, or a pharma- ceutically acceptable salt or solvate thereof, wherein R2 is C1-C6 alkyl substituted with one or more -OH groups, or C1-C6 alkyl substituted with one or more halo groups.

6. The pharmaceutical composition according to claim 3, or a pharma- ceutically acceptable salt or solvate thereof, wherein R2 is unsubstituted C1-C6 alkyl.

7. The pharmaceutical composition of claim 3, or a pharma- ceutically acceptable salt or solvate thereof, wherein R2 is phenyl optionally substituted with one or more of halo, cyano, -OR5, -NR5R6, -S(O)2R5, or -S(O)2OR5.

8. The pharmaceutical composition of claim 3, or a pharma- ceutically acceptable salt or solvate thereof, wherein R2 is -C1-C6 alkyl-phenyl optionally substituted with one or more of halo, cyano, -OR5, -NR5R6, -S(O)2R5, or -S(O)2OR5.

9. The pharmaceutical composition of claim 1, or a pharma- ceutically acceptable salt or solvate thereof, wherein R3 and R4 are independently selected from -F, -Cl, and -Br.

10. The pharmaceutical composition according to claim 9, or a pharma- ceutically acceptable salt or solvate thereof, wherein both of R3 and R4 are -Br.

11. The pharmaceutical composition according to claim 9, or a pharma- ceutically acceptable salt or solvate thereof, wherein both of R3 and R4 are -Cl.

12. The fatty acid amide hydrolase (FAAH) cleavable prodrug of formula (I'), 【Chemistry 2】 【Chemistry 3】 【Chemistry 4】 【Chemistry 5】 【Chemistry 6】 【Chemistry 7】 【Chemistry 8】 2. The pharmaceutical composition of claim 1, having a structure selected from:

13. The pharmaceutical composition according to any one of claims 1 to 12, or a pharma- ceutically acceptable salt or solvate thereof, wherein the peripherally restricted FAAH inhibitor is ASP-3652.

14. Use of a pharmaceutical composition according to any one of claims 1 to 12, or a pharma- ceutically acceptable salt or solvate thereof, in the manufacture of a medicament for treating a CNS disease or disorder in a patient in need of such treatment.

15. The CNS disease or disorder is acute disseminated encephalomyelitis (ADEM), acute hemorrhagic leukoencephalitis (AHL or AHLE), adult Refsum disease, infantile Refsum disease, Alexander disease, Alzheimer's disease, Barrow concentric sclerosis, Canavan disease, central pontine myelinolysis (CPM), cerebral palsy, cerebrotendinous xanthomatosis, chronic inflammatory demyelinating polyneuropathy (CIDP), Devic's syndrome, diffuse spinal fragmentation sclerosis, encephalomyelitis, Guillain-Barré syndrome, idiopathic inflammatory demyelinating disease (HDD), Krabbe disease, or Leber's hereditary optic atrophy.

15. The use according to claim 14, wherein the disease is selected from the group consisting of myelodysplastic syndromes, leukodystrophy, Marburg multiple sclerosis, Marchiafava-Bienami disease, metachromatic leukodystrophy (MLD), multifocal motor neuropathy (MMN), multiple sclerosis (MS), paraproteinaceous demyelinating polyneuropathy, Pelizaeus-Merzbach disease (PMD), progressive multifocal leukoencephalopathy (PML), tropical spastic paraparesis (TSP), X-linked adrenoleukodystrophy (X-ALD, ALO, or X-linked ALO) and Zellweger syndrome.

16. Use of the pharmaceutical composition of claim 13, or a pharma- ceutically acceptable salt or solvate thereof, in the manufacture of a medicament for treating a CNS disease or disorder in a patient in need of treatment.

17. The CNS disease or disorder is acute disseminated encephalomyelitis (ADEM), acute hemorrhagic leukoencephalitis (AHL or AHLE), adult Refsum disease, infantile Refsum disease, Alexander disease, Alzheimer's disease, Barrow concentric sclerosis, Canavan disease, central pontine myelinolysis (CPM), cerebral palsy, cerebrotendinous xanthomatosis, chronic inflammatory demyelinating polyneuropathy (CIDP), Devic's syndrome, diffuse spinal fragmentation sclerosis, encephalomyelitis, Guillain-Barré syndrome, idiopathic inflammatory demyelinating disease (HDD), Krabbe disease, or Leber's hereditary optic atrophy.

17. The use according to claim 16, wherein the disease is selected from the group consisting of myelodysplastic syndromes, leukodystrophy, Marburg multiple sclerosis, Marchiafava-Bienami disease, metachromatic leukodystrophy (MLD), multifocal motor neuropathy (MMN), multiple sclerosis (MS), paraproteinaceous demyelinating polyneuropathy, Pelizaeus-Merzbach disease (PMD), progressive multifocal leukoencephalopathy (PML), tropical spastic paraparesis (TSP), X-linked adrenoleukodystrophy (X-ALD, ALO, or X-linked ALO) and Zellweger syndrome.

Images