Trans-4-hydroxycyclohexylphenylamide mitofusin activator and method of use thereof

Abstract

[Solution] Compounds and compositions containing stereoisomers of 6-phenylhexanamide derivative small molecule mitofusin activators are described. In particular, mitofusin activators containing derivatives of (trans-4-hydroxycyclohexyl)-6-phenylhexanamide are described that are useful for treating diseases or disorders associated with mitochondrial-related diseases, disorders, or conditions, such as diseases or disorders associated with mitofusin-1 (MFN1) and / or mitofusin-2 (MFN2), or mitochondrial dysfunction. Methods of treatment and pharmaceutical formulations are also described.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 62 / 797,513, filed January 28, 2019, and U.S. Provisional Application No. 62 / 949,060, filed December 17, 2019.

[0002] Various aspects of the present disclosure include the provision of novel active trans-4-hydroxycyclohexylphenylamide stereoisomers of small molecule mitofusin activators and methods of use thereof.

[0003] The present disclosure relates generally to compositions and methods for treating inherited or acquired central and peripheral neuropathies, neurodegenerative diseases, disorders or conditions. Summary of the Invention

[0004] Various aspects of the present disclosure include the provision of novel functionally active stereoisomers of small molecule mitofusin activators and methods of use thereof.

[0005] One aspect of the present disclosure provides methods for treating a neuropathy, neurodegenerative disease, neurodegenerative disorder, or neurodegenerative condition. In some embodiments, the methods comprise administering to a subject a therapeutically effective amount of a composition comprising one or more active stereoisomers of a mitofusin activator, wherein the mitofusin activator stimulates mitochondrial fusion and mitochondrial intracellular transport in neurons, thereby inducing resistance to neuronal injury, accelerating neuronal repair, and promoting neuronal regrowth and regeneration.

[0006] Another aspect of the present disclosure provides methods for activating mitofusin in a subject in need thereof. In some embodiments, the methods comprise administering to a human subject or animal patient having a genetic or acquired central or peripheral neuropathy, neurodegenerative disease, disorder, or condition a composition comprising one or more active trans-4-hydroxycyclohexyl stereoisomers of a mitofusin activator, wherein the active trans-4-hydroxycyclohexyl stereoisomers of the mitofusin activator stimulate mitochondrial fusion and mitochondrial intracellular transport in neurons, thereby inducing resistance to neuronal injury, accelerating neuronal repair, and promoting neuronal regrowth and regeneration.

[0007] Another aspect of the present disclosure provides a method for preventing, alleviating, reducing, or enhancing recovery from iatrogenic nerve injury, traumatic nerve injury, or collateral nerve injury in a subject in need thereof. In some aspects, the method comprises administering to the subject a composition comprising one or more active trans-4-hydroxycyclohexyl stereoisomers of a mitofusin activator, wherein the active stereoisomers of the mitofusin activator stimulate mitochondrial fusion and mitochondrial intracellular transport in neurons, thereby inducing resistance to neuronal injury, accelerating neuronal repair, and promoting neuronal regrowth and regeneration, wherein the subject or affected animal has a genetic or acquired central or peripheral neuropathy, neurodegenerative disease, disorder, or condition.

[0008] In some embodiments, the active trans-4-hydroxycyclohexyl stereoisomer of mitofusin activators has substantially better functional potency than both 1-[2-(benzylsulfanyl)ethyl]-3-(2-methylcyclohexyl)urea (Cpd A, Rocha Science 2018) and 2-{2-[(5-cyclopropyl-4-phenyl-4H-1,2,4-triazol-3-yl)sulfanyl]propanamide}-4H,5H,6H-cyclopenta[b]thiophene-3-carboxamide (Cpd B, Rocha Science 2018).

[0009] In some embodiments, the active trans-4-hydroxycyclohexyl stereoisomer of a mitofusin activator has substantially better drug-like pharmacokinetic properties than both 1-[2-(benzylsulfanyl)ethyl]-3-(2-methylcyclohexyl)urea (Cpd A, Rocha Science 2018) and 2-{2-[(5-cyclopropyl-4-phenyl-4H-1,2,4-triazol-3-yl)sulfanyl]propanamide}-4H,5H,6H-cyclopenta[b]thiophene-3-carboxamide (Cpd B, Rocha Science 2018).

[0010] In some embodiments, the active trans-4-hydroxycyclohexyl stereoisomer of a mitofusin activator targets mitofusin-1 (MFN1) or mitofusin-2 (MFN2), increases mitochondrial elongation by enhancing mitochondrial fusion, enhances mitochondrial function as measured by electrochemical polarization of the inner membrane, enhances mitochondrial transport in neuronal axons, corrects cellular and organ dysfunction caused by primary or secondary mitochondrial abnormalities, and corrects mitochondrial defects (e.g., dysmorphomes). reverse mitochondrial fusion, function, tethering, transport, trafficking (e.g., axonal mitochondrial transport), mobility or movement (e.g., loss of polarity, loss of motility); restore, activate, control, regulate, promote, or enhance mitochondrial (as appropriate, mitochondria in nerves or neurons) fusion, function, tethering, trafficking (e.g., axonal mitochondrial transport), mobility or movement; increase mitochondrial elongation or mitochondrial aspect ratio; disrupt intramolecular constraints in MFN2; allosterically activate MFN2; and repair morphological and functional defects in diseased or damaged neurons with mitochondrial abnormalities.

[0011] In some embodiments, the active trans-4-hydroxycyclohexyl stereoisomer mitofusin activator has formula (I): [ka] or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein R 1 is unsubstituted, monosubstituted or polysubstituted C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 Aryl or C 3-8 It may be a heterocyclyl.

[0012] In some embodiments, the active trans-4-hydroxycyclohexyl stereoisomer mitofusin activator has the formula (I): [ka] wherein R 1 The following part: [ka] It can be one of:

[0013] In some embodiments, R 1 is acetamide, C 1-8 Alkoxy, amino, azo, Br, C 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 and optionally substituted with one or more of heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfur dioxide, and / or thiophene, and optionally further substituted with one or more of acetamido, alkoxy, amino, azo, Br, C 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 Optionally, the above-mentioned alkyl, cycloalkyl, heteroaryl, heterocyclyl, indole or phenyl may be substituted with the following: acetamido, alkoxy, amino, azo, Br, C. 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 It may be further substituted with one or more of heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfur dioxide and / or thiophene.

[0014] In some embodiments, Formula (I) comprises the following moiety: [ka] N-(cis-4-hydroxycyclohexyl)-6-phenylhexanamide, [ka] N-(trans-4-hydroxycyclohexyl)-6-phenylhexanamide, [ka] N-(trans-4-hydroxycyclohexyl)-5-phenoxypentanamide, [ka] 4-(benzyloxy)-N-(trans-4-hydroxycyclohexyl)-4-butanamide, [ka] N-(trans-4-hydroxycyclohexyl)-3-phenethoxypropanamide, [ka] N-(trans-4-hydroxycyclohexyl)-2-(3-phenylpropoxy)acetamide, and [ka] 4-Phenylbutyl-(trans-4-hydroxycyclohexyl)carbamate It can be one of:

[0015] Yet another aspect of the present disclosure provides pharmaceutical compositions comprising an active trans-4-hydroxycyclohexyl stereoisomer mitofusin activator, optionally together with one or more therapeutically acceptable diluents or carriers.

[0016] In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable excipient.

[0017] In some embodiments, the pharmaceutical composition comprises one or more of the following: a neuroprotective agent, an anti-Parkinson's agent, an amyloid protein deposition inhibitor, a beta-amyloid synthesis inhibitor, an antidepressant, an anti-anxiety agent, an anti-psychotic agent, an anti-amyotrophic lateral sclerosis agent, an anti-Huntington's disease agent, an anti-Alzheimer's agent, an anti-epileptic agent, and / or a steroid.

[0018] Yet another aspect of the present disclosure provides a method of treating a mitochondrial-related disease, disorder, or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of a mitofusin activator.

[0019] In some embodiments, the subject may be diagnosed with or suspected of having a mitochondrial-related disease, disorder, or condition.

[0020] In some embodiments, the mitochondrial-related disease, disorder, or condition is a central nervous system (CNS) or peripheral nervous system (PNS) injury or trauma, such as trauma to the CNS or PNS, crush injury, spinal cord injury (SCI), traumatic brain injury, stroke, optic nerve injury, or related conditions involving axonal transection, a chronic neurodegenerative condition in which mitochondrial fusion, health, or trafficking is impaired, a disease or disorder associated with mitofusin 1 (MFN1) or mitofusin 2 (MFN2) or mitochondrial dysfunction, fragmentation, or fusion, unfolding of MFN1 or MFN2, or mitochondrial dysfunction, fragmentation, or fusion. Mitochondrial dysfunction due to mutations, degenerative neurological conditions such as Alzheimer's disease, Parkinson's disease, Charcot-Marie-Tooth disease or Huntington's disease, hereditary motor and sensory neuropathies, autism, autosomal dominant optic atrophy (ADOA), muscular dystrophies, Lou Gehrig's disease, cancer, mitochondrial myopathies, diabetes mellitus with hearing loss (DAD), Leber's hereditary optic neuropathy (LHON), Leigh's syndrome, subacute sclerosing encephalopathy, neuropathy, ataxia, retinitis pigmentosa, and ptosis (NARP), myoneurogenic gastrointestinal encephalopathy The primary cause of the mitochondrial myopathy may be one or more of the following: mitochondrial neurogastrointestinal encephalopathy (MNGIE), myoclonic epilepsy with ragged-red fibers syndrome (MERRF), mitochondrial myopathy, encephalomyopathy, lactic acidosis, and stroke-like events (MELAS), mtDNA depletion, mitochondrial neurogastrointestinal encephalopathy (MNGIE), autonomic mitochondrial myopathy, mitochondrial channelopathy, and / or pyruvate dehydrogenase complex deficiency (PDCD / PDH).

[0021] Other objects and features will be in part apparent and in part pointed out hereinafter. [Brief explanation of the drawings]

[0022] Those skilled in the art will understand that the drawings described below are for illustrative purposes only and are not intended to limit the scope of the present teachings in any way.

[0023] [Figure 1] Figure 1A illustrates structural models of human MFN2 in the closed / inactive conformation (left) and the open / active conformation (right) that can be promoted by mitofusin activators. Figure 1B shows pharmacophore modeling of interacting amino acid side chains that led to prototypical mitofusin small molecule agonists (Rocha, et al.; Science 2018).

[0024] [Figure 2] Figures 2A-C show the structure-function relationship of the cis- and trans-stereoisomers of MiM111 (see, e.g., Example 3). Figure 2A shows dose-response curves for the cis-(Cpd15A) and trans-(Cpd15B) MiM111 stereoisomers in cells expressing only MFN2. Figure 2B shows dose-response curves for the cis-(Cpd15A) and trans-(Cpd15B) MiM111 stereoisomers in cells expressing only MFN1. "Cpd2" corresponds to the comparative prototype mitofusin activator, Chimera C. Figure 2C shows FRET analysis of the conformational switch induced by the mitofusin activator. The open conformation is more active. "Peptide" is the MP1 mitofusin agonist peptide described in reference Franco Nature 2016 and shown in Figure 1A.

[0025] [Figure 3] 3A-3B show the chemical structures and corresponding NMR spectra of the cis- (FIG. 3A) and trans- (FIG. 3B) stereoisomers of MiM111 (see, e.g., Example 3). Black arrows and arrowheads are specific to the cis-isomer, while gray arrowheads represent the trans-isomer.

[0026] [Figure 4]Figures 4A-4B show the in vivo (mouse) pharmacokinetic properties of trans-MiM111 (Cpd15B) (see, e.g., Example 5). Figure 4A shows total plasma and brain concentrations after a single IV injection. Figure 4B shows steady-state elimination kinetics after 3 days of continuous subcutaneous infusion.

[0027] [Figure 5] Figures 5A-5B show the oral bioavailability and in vivo target association of trans-MiM111 (Cpd15B) (see, e.g., Example 6). Figure 5A shows plasma levels after a single IV dose (filled circles) or oral administration (open circles). Figure 5B shows a kymograph showing mitochondrial motility in the sciatic nerve of a CMT2A mouse 6 hours after oral administration as in A. Motile mitochondria exhibit horizontal movement (quantitative data for the corresponding groups are on the right). DETAILED DESCRIPTION OF THE INVENTION

[0028] The present disclosure is based at least on the discovery that pharmacophore modeling of MFN2-derived interacting peptides essential for function can generate structurally diverse small molecule peptidomimetic activators useful for treating mitochondrial-related diseases, disorders, and conditions. As demonstrated herein, the present disclosure provides stereoisomer-specific chemical entities or compositions for controlling mitochondrial function. These compositions may be useful for correcting organelle, cellular, and organ dysfunction due to primary or secondary mitochondrial abnormalities that cause or contribute to disease pathology and dysfunction.

[0029] Mitofusin activator

[0030] The present disclosure provides a class of stereoisomer-specific trans-4-hydroxycyclohexyl derivative small molecules that promote a conformational change in MFN1 and MFN2, resulting in enhanced activation. As described herein, compositions for treating mitochondrial-related diseases, disorders, or conditions can include active trans-4-hydroxycyclohexyl mitofusin activators, such as peptidomimetics (e.g., small molecules that mimic the chemical structural features of peptides). The peptidomimetics can be chemical peptidomimetics. For example, the peptidomimetics can mimic minipeptides derived from mitofusin.

[0031] This disclosure describes the functional activity of small molecule peptidomimetics, which requires mimicking the intrinsic peptide conformation in three-dimensional space. Diastereomers of the same chemical mitofusin activator prefer different three-dimensional structures (Figures 1A-1B). Accordingly, cis- and trans-4-hydroxycyclohexyl derivatives differ significantly in their ability to physically associate with and functionally activate their mitofusin protein targets (Figures 2A-2C).

[0032] As described herein, a new generation of trans-4-hydroxycyclohexyl derivative peptidomimetic small molecules has been developed that exhibit stereoisomer-specific functional activity. These compounds activate mitochondrial fusion by inducing MFN1 and MFN2 into distinct conformational states. Prototype small molecule peptidomimetics targeting MFN1 or MFN2 (described in Rocha, et al., Science, 2018) have not led to drug development due to poor pharmacokinetic properties. Described herein are active stereoisomers of a structurally distinct class of small molecule mitofusin activators that activate mitochondrial fusion and intracellular trafficking, have favorable pharmacokinetic properties, and can be used to correct mitochondrial and cellular dysfunction.

[0033] Mitofusin minipeptide

[0034] As described herein, peptide mitofusin activators can be minipeptides derived from MFN2 as described in Franco, et al.; Nature 2016.

[0035] MFN activator (fusion-promoting) peptide mimetics

[0036] As described herein, the peptidomimetic can be an MFN activator (fusion-promoting) peptidomimetic that competes with endogenous MFN1 or MFN2 HR1-HR2 peptide-peptide interactions, such as those described in Franco, et al.; Nature 2016 and Rocha, et al.; Science 2018.

[0037] The prototypical mitofusin activator according to the present disclosure, Chimera C, is a compound comprising the following: [ka] 1-(3-(5-cyclopropyl-4-phenyl-4H-1,2,4-triazol-3-yl)propyl)-3-(2-methylcyclohexyl)urea (Chimera C, MW: 381.52 g / mol, formula: C 22 H 31 N5O).

[0038] Mitofusin activators: structurally distinct small molecules that activate MFN1 and / or MFN2

[0039] The small molecule mitofusin activators described herein were designed in part using the pharmacophore HR1-HR2 peptide-peptide interaction model described in Rocha, et al.; Science 2018, but are structurally distinct from the chemical class described by Rocha and are a distinct chemical class of allosteric mitofusin activators. Activators are substances that partially or fully activate proteins to which they bind.

[0040] The mitofusin activator has the formula (I): [ka] or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein R 1 The following part: [ka] may be selected from:

[0041] If necessary, R in formula (I) 1 are independently selected from the following groups: acetamido, C 1-8 Alkoxy, amino, azo, Br, C 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 and optionally further substituted by one or more of heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfur dioxide and / or thiophene, and optionally further substituted by acetamido, alkoxy, amino, azo, Br, C 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 Heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfur dioxide and / or thiophene, and alkyl, cycloalkyl, heteroaryl, heterocyclyl, indole or phenyl may optionally further be substituted with the following groups: acetamido, alkoxy, amino, azo, Br, C 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 It may be substituted with one or more of heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide and / or thiophene.

[0042] If necessary, R in formula (I)1 The groups may be independently selected from the following groups: hydroxyl, C 1-10 Alkyl hydroxyl, amine, C 1-10 Carboxylic acid, C 1-10 Carboxyl, linear or branched C optionally containing unsaturation 1-10 Alkyl, optionally unsaturated or containing one oxygen or nitrogen atom C 2-8 Cycloalkyl, straight or branched chain C 1-10 The unsubstituted or substituted phenyl rings may be independently substituted with one or more of the following groups: alkylamine, heterocyclyl, heterocyclicamine, and / or aryl containing phenyl and heteroaryl containing 1 to 4 of the following heteroatoms: N, O, and / or S, unsubstituted phenyl rings, substituted phenyl rings, unsubstituted heterocyclyl, and substituted heterocyclyl. Optionally, the unsubstituted or substituted phenyl rings may be independently substituted with one or more of the following groups: hydroxyl, C 1-10 Alkyl hydroxyl, amine, C 1-10 Carboxylic acid, C 1-10 Carboxyl, linear or branched C optionally containing unsaturation 1-10 Alkyl, straight or branched chain optionally containing unsaturation 1-10 Alkylamines, optionally unsaturated or containing one oxygen or nitrogen atom, C 2-10 Cycloalkyl, straight or branched chain C 1-10 The heterocyclyl may be substituted with one or more of the following groups: alkylamine, heterocyclyl, heterocyclicamine, and / or aryl, including phenyl, and heteroaryl, including 1 to 4 of the following heteroatoms: N, O, and / or S. Optionally, the unsubstituted or substituted heterocyclyl may independently be substituted with one or more of the following groups: hydroxyl, C 1-10 Alkyl hydroxyl, amine, C 1-10 Carboxylic acid, C 1-10 Carboxyl, linear or branched C optionally containing unsaturation 1-10 Alkyl, straight or branched chain optionally containing unsaturation 1-10 Alkylamines, optionally unsaturated or containing one oxygen or nitrogen atom, C 2-8 Cycloalkyl, heterocyclyl, straight or branched chain C 1-10They may be substituted with one or more of alkylamines, heterocyclic amines, and / or aryls containing phenyl and heteroaryls containing 1 to 4 of the following heteroatoms: N, O, and S. Any of the above may be further substituted as needed.

[0043] In some embodiments, R in formula (I) 1 optionally containing the following groups: acetamido, alkoxy, amino, azo, Br, C 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 and optionally further substituted by one or more of the following groups: heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfur dioxide and / or thiophene, and optionally further substituted by one or more of the following groups: acetamido, alkoxy, amino, azo, Br, C 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 and alkyl, cycloalkyl, heteroaryl, heterocyclyl, indole, or phenyl may optionally be further substituted with one or more of acetamido, alkoxy, amino, azo, Br, C. 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 It may be substituted with one or more of heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfur dioxide and / or thiophene.

[0044] In another embodiment of the present disclosure, the mitofusin activator has the formula (II): [ka] or a pharmaceutically acceptable salt thereof. In formula (II), o can be 0, 1, 2, 3, 4, or 5, p can be 0 or 1, and q can be 0, 1, 2, 3, 4, or 5, provided that the sum of o + p + q is 3 or more or 7 or less, Z can be cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, and R 2 and R 3 are independently H, F, alkyl or C 3-7 R may be cycloalkyl; 2 and R 3 Together, C 3-7 may form a cycloalkyl or heterocycloalkyl, R 4 and R 5 are independently H, F, alkyl, COR 7 , C 3-7 cycloalkyl or optionally R 4 and R 5 Together, C 3-7 It can form a cycloalkyl or heterocycloalkyl, and Y can be O, CR 5 R 6 , C.R. 7 =CR 8 , a triple bond, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, provided that when o is 1 or more, Y=NR 7 ,S,SO2,SONR 8 , -NR 8 SO2-, -NR 7 CO-, -CONR 7 -, -NR 7 CONR 8 - and each R 7 are independently H, alkyl and C 3-7 cycloalkyl, and each R 8 are independently H, alkyl, COR 7 and C 3-7 cycloalkyl, or optionally R 7 and R 8 Together, C 3-7 It can form a cycloalkyl.

[0045] In some embodiments, in the mitofusin activator of formula (II), o can be 0, 1, 2, 3, 4, or 5; p can be 0 or 1; q can be 0, 1, 2, 3, 4, or 5, provided that the sum of o + p + q is 3 or more or 7 or less; Z can be cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; Y can be O, CR 5 R 6 , cycloalkyl, or aryl; R 2 , R 3 , R 4 , R 5 may be independently selected from H or alkyl.

[0046] In some embodiments, in the mitofusin activator of formula (II), o can be 0, 1, 2, 3, 4, or 5; p can be 0 or 1; q can be 0, 1, 2, 3, 4, or 5, provided that the sum of o + p + q is 3 or more or 5 or less; Z can be aryl or heteroaryl; Y can be O, CH, or cycloalkyl; R 2 , R 3 , R 4 and R 5 can each be H.

[0047] In some embodiments, in the mitofusin activator of formula (II), o can be 0, 1, 2, or 3, p can be 1, and q can be 0, 1, 2, or 3, provided that the sum of o+p+q is 3 or more or 5 or less, Z can be aryl or heteroaryl, Y can be cyclopropyl or cyclobutyl, and R 2 , R 3 , R 4 and R 5 can each be H.

[0048] In some embodiments, in the mitofusin activator of formula (II), Z can be aryl or heteroaryl, Y can be O or CH, and R 2 , R 3 , R 4 and R 5may each be H, o may be 0, 1, 2, 3, or 4, p may be 1, and q may be 0, 1, 2, 3, or 4, provided that the sum of o+p+q is 5.

[0049] In some embodiments, in the mitofusin activator of formula (II), Z can be phenyl or heteroaryl, where the heteroaryl can contain 1 to 4 heteroatoms independently selected from N, O, and S, and the phenyl or heteroaryl can be selected from the following independently selected substituents: R 7 , OR 7 , Cl, F, -CN, CF3, -NR 7 R 8 , -SO2NR 7 R 8 , -NR 7 SO2R 9 , -SO2R 9 , -CONR 7 R 8 , -NR 7 COR 9 , C 3-7 cycloalkyl and / or heterocycloalkyl, and each R 7 are independently H, alkyl and C 3-7 cycloalkyl, and each R 8 are independently H, alkyl, COR 7 and C 3-7 cycloalkyl, or optionally R 7 and R 8 Together, C 3-7 can form a cycloalkyl, Y can be O or CH, and R 1 , R 2 , R 3 and R 4 may each be H, o may be 0, 1, 2, 3, or 4, p may be 1, and q may be 0, 1, 2, 3, or 4, provided that the sum of o+p+q is 5.

[0050] In some embodiments, in the mitofusin activator of formula (II), Z can be phenyl or heteroaryl, which can contain 1 to 3 of the following heteroatoms: N, O, and S, and the phenyl or heteroaryl can be R 7 , OR 7 , Cl, F, -CN, CF3, -NR 7 R 8 , -SO2R 9 , -CONR 7 R 8 , -NR 7 COR 9 , C 3-7 and R may contain 0 to 3 substituents independently selected from cycloalkyl and / or heterocycloalkyl, Y may be O or CH2, and R 1 , R 2 , R 3 and R 4 can each be H, and each R 7 are independently H, alkyl and C 3-7 cycloalkyl, and each R 8 are independently H, alkyl, COR 7 and C 3-7 cycloalkyl, or optionally R 7 and R 8 Together, C 3-7 can form a cycloalkyl, and each R 9 is alkyl or C 3-7 cycloalkyl, where o can be 0, 1, 2, 3, or 4, p can be 1, and q can be 0, 1, 2, 3, or 4, provided that the sum of o+p+q is 5.

[0051] In some embodiments, in the mitofusin activator of formula (II), X can be 4-hydroxylcyclohexyl, 4-aminocyclohexyl, 4-(N-methyl)aminocyclohexyl, 4-(N,N-dimethyl)aminocyclohexyl, 4-(N-acetylamino)cyclohexyl, 4,4-difluorocyclohexyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, 4-N-methyl-piperidinyl, or 4-N-acetyl-piperidinyl; Z can be phenyl, 2-pyridinyl, 3-pyridinyl, 4-pyridinyl, 6-pyrimidinyl, 5-pyrimidinyl, 4-pyrimidinyl, or 2-pyrimidinyl; and the phenyl, pyridinyl, and pyrimidinyl moieties can independently be selected from the following independently selected substituents: R 7 , OR 7 , Cl, F, -CN, CF3, -NR 7 R 8 , -SO2R 9 , -CONR 7 R 8 and / or -NR 7 COR 9 Y may be O or CH2, and R 1 , R 2 , R 3 and R 4 can each be H, and each R 7 are independently H, alkyl and C 3-7 cycloalkyl, and each R 8 are independently H, alkyl, COR 7 and C 3-7 cycloalkyl, or optionally R 7 and R 8 Together, C 3-7 can form a cycloalkyl, R 9 is alkyl or C 3-7 cycloalkyl, where o can be 0, 1, 2, 3, or 4, p can be 1, and q can be 0, 1, 2, 3, or 4, provided that the sum of o+p+q is 5.

[0052] In another embodiment of the present disclosure, a method for treating a disease exhibiting a need for a mitofusin activator comprises administering to a mammal in need thereof a therapeutically effective amount of a compound of formula (II) [ka] or a pharmaceutically acceptable salt thereof. In formula (II), o can be 0, 1, 2, 3, 4, or 5, p can be 0 or 1, and q can be 0, 1, 2, 3, 4, or 5, provided that the sum of o + p + q is 3 or more or 7 or less, Z can be cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, and R 2 and R 3 are independently H, F, alkyl or C 3-7 cycloalkyl or optionally R 2 and R 3 Together, C 3-7 can form a cycloalkyl or heterocycloalkyl, R 4 and R 5 are independently H, F, alkyl, COR 7 and / or C. 3-7 cycloalkyl or optionally R 4 and R 5 Together, C 3-7 It can form a cycloalkyl or heterocycloalkyl, and Y can be O, CR 6 R 7 , C.R. 8 =CR 9 , a triple bond, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, provided that when o is 1 or greater, Y is NR 8 ,S,SO2,SONR 9 , -NR 9 SO2-, -NR 8 CO-, -CONR 8 -or-NR 8 CONR 9 - and R 6 and R 7 may independently be H, F, alkyl and / or cycloalkyl, or optionally R 6 and R7 Together, C 3-7 can form a cycloalkyl or heterocycloalkyl, R 8 and R 9 are independently H, alkyl and / or C 3-7 It may be cycloalkyl.

[0053] In some embodiments, in the methods of treating a disease indicating a need for a mitofusin activator, the PNS or CNS disorder is a chronic neurodegenerative condition in which mitochondrial fusion, health, or trafficking is impaired, a disease or disorder associated with dysfunction of mitofusin-1 (MFN1) or mitofusin-2 (MFN2), a disease associated with mitochondrial fragmentation, dysfunction, or movement dysfunction, a degenerative neuromuscular condition, e.g., Charcot-Marie-Tooth disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, Alzheimer's disease, Parkinson's disease, hereditary motor and sensory neuropathies, autism, autosomal dominant optic neuropathy (ADOA), muscular dystrophy, Lou Gehrig's disease, cancer, mitochondrial myopathy, diabetes mellitus with hearing loss (DAD), Leber's syndrome, or a combination thereof. The neuropathy may be selected from any one or combination of: Transmissible Optic Neuropathy (LHON), Leigh's Syndrome, Subacute Sclerosing Encephalopathy, Neuropathy, Ataxia, Retinitis Pigmentosa, and Ptosis (NARP), Myoneurogastrointestinal Encephalopathy (MNGIE), Myoclonic Epilepsy with Ragged-Red Fibers Syndrome (MERRF), Mitochondrial Myopathy, Encephalomyopathy, Lactic Acidosis, and Stroke-like Events (MELAS), mtDNA depletion, Mitochondrial Neurogastrointestinal Encephalopathy (MNGIE), Autonomic Mitochondrial Myopathy, Mitochondrial Channelopathies or Pyruvate Dehydrogenase Complex Deficiency (PDCD / PDH), Diabetic Neuropathy, Chemotherapy-Induced Peripheral Neuropathy, Crush Injury, SCI, Traumatic Brain Injury (TBI), Stroke, Optic Nerve Injury, and / or Related Conditions with Axonal Transcription.

[0054] In some embodiments of the present disclosure, in methods of treating diseases exhibiting a need for a mitofusin activator, the composition may further comprise a pharmaceutically acceptable excipient.

[0055] In some embodiments of the present disclosure, a method for treating genetic and / or non-genetic neurodegenerative conditions, injuries, damage and / or trauma of the CNS and / or PNS comprises administering to a subject a therapeutically effective amount of a mitofusin activator of the present disclosure.

[0056] In some embodiments of the present disclosure, in methods for treating hereditary or non-hereditary neurodegenerative conditions, injuries, damage, or trauma of the CNS or PNS, the subject is treated with one or more of the following: a chronic neurodegenerative condition in which mitochondrial fusion, health, or trafficking is impaired; a disease or disorder associated with dysfunction of MFN1 or MFN2; a disease associated with mitochondrial fragmentation, dysfunction, or movement dysfunction; a degenerative neuromuscular condition (e.g., Charcot-Marie-Tooth disease, ALS, Huntington's disease, Alzheimer's disease, Parkinson's disease); a hereditary motor and sensory neuropathic disorder; May be diagnosed with or suspected of having one or more of the following conditions: autism, ADOA, muscular dystrophy, Lou Gehrig's disease, cancer, mitochondrial myopathy, DAD, LHON, Leigh's syndrome, subacute sclerosing encephalopathy, NARP, MNGIE, MERRF, MELAS, mtDNA depletion, MNGIE, autonomic neuropathic mitochondrial myopathy, mitochondrial channelopathy, PDCD / PDH, diabetic neuropathy, chemotherapy-induced peripheral neuropathy, crush injury, SCI, TBI, stroke, optic nerve injury, and / or related conditions involving axonal transection.

[0057] The term "imine" or "imino," as used herein, unless otherwise indicated, includes functional groups or compounds containing a carbon-nitrogen double bond. The phrase "imino compound," as used herein, refers to a compound containing an "imine" or "imino" group, as defined herein, unless otherwise indicated. The "imine" or "imino" group can be optionally substituted.

[0058] As used herein, the term "hydroxyl" includes --OH unless otherwise indicated. "Hydroxyl" can be optionally substituted.

[0059] The terms "halogen" and "halo" as used herein include chlorine, chloro, Cl, fluorine, fluoro, F, bromine, bromo, Br, and iodine, iodo, or I, unless otherwise indicated.

[0060] As used herein, the term "acetamide" is an organic compound having the formula CH3CONH2. "Acetamide" can be optionally substituted.

[0061] As used herein, the term "aryl" includes carbocyclic aromatic groups unless otherwise specified. Examples of aryl groups include, but are not limited to, phenyl, benzyl, naphthyl, and anthracenyl. An "aryl" can be optionally substituted.

[0062] As used herein, unless otherwise indicated, the terms "amine" and "amino" include functional groups containing a nitrogen atom having a lone pair of electrons, in which one or more hydrogen atoms have been replaced by a substituent, such as, but not limited to, an alkyl or aryl group. An "amine" or "amino" group can be optionally substituted.

[0063] The term "alkyl," as used herein, unless otherwise indicated, includes monovalent saturated hydrocarbon radicals having straight-chain or branched-chain moieties, such as, but not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, and octyl groups. Representative straight-chain lower alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl. Branched lower alkyl groups include, but are not limited to, isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, 2-methylbutyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, 3,3-dimethylpentyl, 2,3,4-trimethylpentyl, 3-methylhexyl, 2,2-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 3,5-dimethylhexyl, 2,4-dimethylpentyl, 2-methylheptyl, and 3-methylheptyl. 1-8 Alkyl includes, but is not limited to, vinyl, allyl, 1-butenyl, 2-butenyl, isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, 1-hexyl, 2-hexyl, 3-hexyl, acetylenyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, or 3-methyl-1 butynyl. Alkyl can be saturated, partially saturated, or unsaturated. "Alkyl" can be optionally substituted.

[0064] The term "carboxyl," as used herein, unless otherwise indicated, includes a functional group containing a carbon atom double-bonded to an oxygen atom and single-bonded to a hydroxyl group (-COOH). "Carboxyl" can be optionally substituted.

[0065] The term "alkenyl," as used herein, unless otherwise indicated, includes alkyl moieties (as defined above) having at least one carbon-carbon double bond, and includes E and Z isomers of said alkenyl moieties. An alkenyl can be partially saturated or unsaturated. An "alkenyl" can be optionally substituted.

[0066] The term "alkynyl," as used herein, unless otherwise indicated, includes an alkyl moiety (as defined above) having at least one carbon-carbon triple bond. An alkynyl can be partially saturated or unsaturated. An "alkynyl" can be optionally substituted.

[0067] The term "acyl," as used herein, unless otherwise indicated, includes functional groups obtained by removal of the hydroxyl (-OH) group from an aliphatic carboxylic acid. "Acyl" can be optionally substituted.

[0068] The term "alkoxyl," as used herein, unless otherwise indicated, includes O-alkyl groups, where alkyl is as defined above and O represents oxygen. Representative alkoxyl groups include -O-methyl, -O-ethyl, -O-propyl, -O-butyl, -O-pentyl, -O-hexyl, -O-heptyl, -O-octyl, -O-isopropyl, -O-sec-butyl, -O-isobutyl, -O-tert-butyl, -O-isopentyl, -O-2-methylbutyl, -O-2-methylpentyl, -O-3-methylpentyl, -O-2,2-dimethylbutyl, -O-2,3-dimethylbutyl, -O- -2,2-Dimethylpentyl, -O-2,3-Dimethylpentyl, -O-3,3-Dimethylpentyl, -O-2,3,4-Trimethylpentyl, -O-3-Methylhexyl, -O-2,2-Dimethylhexyl, -O-2,4-Dimethylhexyl, -O-2,5-Dimethylhexyl, -O-3,5-Dimethylhexyl, -O-2,4-Dimethylpentyl, -O-2-Methylheptyl, -O-3-Methylheptyl, -O-Vinyl, -O-Allyl, -O-1-Butenyl , -O-2-butenyl, -O-isobutylenyl, -O-1-pentenyl, -O-2-pentenyl, -O-3-methyl-1-butenyl, -O-2-methyl-2-butenyl, -O-2,3-dimethyl-2-butenyl, -O-1-hexyl, -O-2-hexyl, -O-3-hexyl, -O-acetylenyl, -O-propynyl, -O-1-butynyl, -O-2-butynyl, -O-1-pentynyl, -O-2-pentynyl, -O-3-methyl-1-butynyl, -O-cyclohexyl Cyclopropyl, -O-cyclobutyl, -O-cyclopentyl, -O-cyclohexyl, -O-cycloheptyl, -O-cyclooctyl, -O-cyclononyl, -O-cyclodecyl, -O-CH2-cyclopropyl, -O-CH2-cyclobutyl, -O-CH2-cyclopentyl, -O-CH2-cyclohexyl, -O-CH2-cycloheptyl, -O-CH2-cyclooctyl, -O-CH2-cyclononyl, -O-CH2-cyclodecyl, -O-(CH2) n -cyclopropyl, -O-(CH2) n -cyclobutyl, -O-(CH2) n -Cyclopentyl, -O-(CH2)n -Cyclohexyl, -O-(CH2) n -Cycloheptyl, -O-(CH2) n -Cyclooctyl, -O-(CH2) n -cyclononyl and / or -O-(CH2) n Examples include, but are not limited to, -cyclodecyl. Alkoxyl can be saturated, partially saturated, or unsaturated. "Alkoxyl" can be optionally substituted. In any of the above examples, n can be from 1 to about 20.

[0069] The term "cycloalkyl" as used herein, unless otherwise indicated, includes non-aromatic, saturated, partially saturated, or unsaturated, monocyclic, or fused, spiro, or non-fused bicyclic or tricyclic hydrocarbons as referred to herein, containing a total of 3 to 10 carbon atoms. Examples of cycloalkyl include -cyclopropyl, -cyclobutyl, -cyclopentyl, -cyclopentadienyl, -cyclohexyl, -cyclohexenyl, -1,3-cyclohexadienyl, -1,4-cyclohexadienyl, -cycloheptyl, -1,3-cycloheptadienyl, -1,3,5-cycloheptatrienyl, -cyclooctyl, and -cyclooctadienyl. 3-10 Examples of -lower alkyl-cycloalkyl groups include, but are not limited to, -CH-cyclopropyl, -CH-cyclobutyl, -CH-cyclopentyl, -CH-cyclopentadienyl, -CH-cyclohexyl, -CH-cycloheptyl, and / or -CH-cyclooctyl. A "cycloalkyl" can be optionally substituted.

[0070] The term "heterocyclyl" (e.g., "heteroaryl"), as used herein, unless otherwise indicated, includes aromatic or non-aromatic cycloalkyl, in which one to four ring carbon atoms are independently substituted with one or more of O, S, and N. Representative examples of heterocycles include, but are not limited to, benzofuranyl, benzothiophene, indolyl, benzopyrazolyl, coumarinyl, isoquinolinyl, pyrrolyl, pyrrolidinyl, thiophenyl, furanyl, thiazolyl, imidazolyl, pyrazolyl, triazolyl, quinolinyl, pyrimidinyl, pyridinyl, pyridonyl, pyrazinyl, pyridazinyl, isothiazolyl, isoxazolyl, (1,4)-dioxane, (1,3)-dioxane, 4,5-dihydro-1H-imidazolyl, and / or tetrazolyl. Heterocyclyls can be substituted or unsubstituted. The heterocyclyl may also be attached at any ring atom (i.e., any carbon atom or heteroatom of the heterocyclic ring). A heterocycle may be saturated, partially saturated, or unsaturated. A "heterocycle" may be optionally substituted.

[0071] The term "indole" as used herein refers to an aromatic heterocyclic organic compound having the formula CHN. Indole has a bicyclic structure containing a six-membered benzene ring fused to a five-membered nitrogen-containing pyrrole ring. "Indole" can be optionally substituted.

[0072] As used herein, unless otherwise indicated, the term "cyano" includes a -CN group. "Cyano" can be optionally substituted.

[0073] The term "alcohol," as used herein, unless otherwise indicated, includes compounds having a hydroxyl functional group (-OH) attached to a carbon atom. In particular, this carbon atom may be saturated with single bonds to three other atoms. An "alcohol" may be optionally substituted. An "alcohol" may be a primary, secondary, or tertiary alcohol.

[0074] The term "solvate" is intended to mean a solvated form of a particular compound that retains the effectiveness of such compound. Examples of solvates include, but are not limited to, compounds of the invention in combination with one or more of water, isopropanol, ethanol, methanol, dimethyl sulfoxide (DMSO), ethyl acetate, acetic acid, or ethanolamine.

[0075] The term "stereoisomer" includes both optical isomers, such as enantiomers or diastereomers (the latter existing due to two or more centers of asymmetry in the molecule), and geometric isomers (cis / trans isomers or diastereomers).

[0076] A composition comprising a trans-stereoisomer 6-phenylhexanamide derivative mitofusin activator of the present disclosure or a pharmaceutically acceptable salt thereof may contain a greater amount of the trans-stereoisomer than the cis-stereoisomer. A method of using a composition comprising a trans-stereoisomer 6-phenylhexanamide derivative mitofusin activator of the present disclosure or a pharmaceutically acceptable salt thereof may be such that the composition contains a greater amount of the trans-stereoisomer than the cis-stereoisomer.

[0077] As used herein, the term "mmol" is intended to mean millimole. As used herein, the terms "equiv" and "eq." are intended to mean equivalent. As used herein, the term "mL" is intended to mean milliliter. As used herein, the term "g" is intended to mean gram. As used herein, the term "kg" is intended to mean kilogram. As used herein, the term "μg" is intended to mean microgram. As used herein, the term "h" is intended to mean hour. As used herein, the term "min" is intended to mean minute. As used herein, the term "M" is intended to mean molar concentration. As used herein, the term "μL" is intended to mean microliter. As used herein, the term "μM" is intended to mean micromolar concentration. As used herein, the term "nM" is intended to mean nanomolar concentration. As used herein, the term "N" is intended to mean a defined concentration. As used herein, the term "amu" is intended to mean atomic mass unit. As used herein, the term "°C" is intended to mean degrees Celsius. As used herein, the term "wt / wt" is intended to mean weight / weight. As used herein, the term "v / v" is intended to mean volume / volume. As used herein, the term "MS" is intended to mean mass spectrometry. As used herein, the term "HPLC" is intended to mean high performance liquid chromatography. As used herein, the term "RT" is intended to mean room temperature. As used herein, the term "e.g.," is intended to mean, for example. As used herein, the term "N / A" is intended to mean not tested or not applicable.

[0078] As used herein, the expression "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic or inorganic salt of a compound of the present invention. Suitable salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and / or pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoic acid) salt). A pharmaceutically acceptable salt may require the inclusion of another molecule, such as an acetate ion, a succinate ion, or other counterion. The counterion can be any organic or inorganic moiety that stabilizes the charge on the parent compound. Furthermore, a pharmaceutically acceptable salt may have more than one charged atom in its structure. If multiple charged atoms are part of the pharmaceutically acceptable salt, it may have multiple counterions. Thus, a pharmaceutically acceptable salt may have one or more charged atoms and / or one or more counterions. As used herein, the phrase "pharmaceutically acceptable solvate" refers to an association of one or more solvent molecules with a compound of the present invention. Examples of solvents that form pharmaceutically acceptable solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and / or ethanolamine. As used herein, the phrase "pharmaceutically acceptable hydrate" refers to a compound of the present invention or a salt thereof that further contains a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces.

[0079] Each of the conditions, diseases, disorders, and conditions described herein, as well as others, may benefit from the compositions and methods described herein. Generally, treating a condition, disease, disorder, or condition includes preventing or delaying the appearance of clinical symptoms in a mammal that may be afflicted with or susceptible to the condition, disease, disorder, or condition but that has not yet experienced or exhibited clinical or asymptomatic symptoms thereof. Treatment can also include inhibiting the condition, disease, disorder, or condition, e.g., arresting or reducing the onset of the disease or at least one clinical or asymptomatic symptom thereof. Furthermore, treatment can include alleviating the disease, e.g., reversing the condition, disease, disorder, or condition, or at least one clinical or asymptomatic symptom thereof. The benefit to a treated subject may be statistically significant, or at least perceptible to the subject or physician.

[0080] Mitofusin 1 and mitofusin 2

[0081] Mitofusins ​​(MFN) 1 and 2, named for their central role in mitochondrial fusion, are attractive drug targets because their regulatory function in regulating mitochondrial dynamics and quality / quantity is disrupted in several neurodegenerative disorders. In particular, genetic mutations that abolish or impair MFN2 function and thus suppress mitochondrial fusion are responsible for Charcot-Marie-Tooth disease type 2A (CMT2A), a rare autosomal dominant neurodegenerative condition for which no disease-modifying treatments currently exist. Furthermore, accumulating experimental evidence supports the critical roles of MFN1 and MFN2 in cardiac disease. Pharmacological approaches that improve mitofusin function have the potential to correct the underlying causes of CMT2A and other cardiac or neurodegenerative disorders caused by mitochondrial dys-dynamism.

[0082] MFN1 and MFN2 form homodimers (MFN1-MFN1 or MFN2-MFN2) or heterodimers (MFN1-MFN2) between mitochondria. This process, called mitochondrial tethering, is the first essential step in mitochondrial fusion, which is essential for cellular metabolic health (Koshiba, T., et al.; Science 305:858-62, 2004). Mitochondrial tethering and fusion depend on the conformational change of the MFN protein from a more closed, resting state to a more open, active state. The conformation of MFN is regulated by intramolecular peptide-peptide interactions (PPIs) between the alpha helices in the stalk region of the protein (Franco, A., et al.; Nature 540:74-79, 2016). In human (h)MFN2, phosphorylation of serine 378 by mitochondrial PINK1 kinase promotes a rigid alpha helix within the interacting peptide, directing the crucial amino acid side chains of Val372, Met376, and His380 toward their interacting partners, Leu727, Leu723, and Lys720, respectively (Rocha, AG, et al.; Science 360:336-41, 2018). Strong PPI promotes a folded protein conformation that reduces the probability of trans-MFN-MFN dimer formation between mitochondria (e.g., unfavorable for mitochondrial tethering / fusion). When serine 378 is not phosphorylated, the alpha helix of the interacting peptide partially unwinds, weakening the PPI and increasing the probability that the protein unfolds, allowing MFN trans-dimerization and subsequent mitochondrial fusion (Rocha, AG, et al.; Science 2018).

[0083] Franco et al. (Franco, A., et al.; Nature 2016) reported a slightly modified 18-amino acid peptide derived from hMFN2 amino acids 367–384 that competitively inhibits intramolecular PPIs in MFN1 and MFN2, thereby promoting a protein conformation favorable for mitochondrial fusion. Understanding the critical interacting amino acids within this activating peptide led to the development of a pharmacophore model and the identification of a prototypical small-molecule mitofusin activator, chimeric BA / I. This chimeric BA / I mimicked the effects of the mitofusin-activating peptide by promoting mitochondrial fusion after topical application to mitofusin-deficient cultured cells and by increasing mitochondrial motility in ex vivo CMT2A neurons (Rocha, A.G., et al.; Science 2018). Clinical application of small-molecule mitofusin activators with drug-like properties holds the promise of being the first disease-modifying treatment for CMT2A and may pave the way for novel therapeutic approaches to improve mitochondrial fusion in multiple diseases in which mitochondrial dynamics are impaired.

[0084] Mitochondrial-related diseases, disorders or conditions

[0085] The present disclosure provides compositions and therapeutic methods for treating mitochondrial-associated diseases, disorders, or conditions, including diseases or disorders associated with MFN1 and / or MFN2 and mitochondrial dysfunction. The mitochondrial-associated disease, disorder, or condition can be a disease primarily caused by or secondarily associated with mitochondrial dysfunction, fragmentation, or loss of fusion, or a disease associated with dysfunction in the catalytic activity or conformational unfolding of MFN1 or MFN2. Mitochondrial dysfunction can be due to genetic mutations in mitofusin or other (nuclear- or mitochondrially-encoded) genes, or can be due to physical, chemical, or environmental insults to the CNS or PNS.

[0086] Mitochondria traverse the cell, fuse, exchange genomes, and promote mutual repair. Mitochondrial fusion and intracellular transport are mediated in part by MFN1 and MFN2. Mutations in MFN2, which inhibit mitochondrial fusion and motility, cause Charcot-Marie-Tooth disease type 2A (CMT2A), the most common inherited axonal neuropathy. Mitochondrial fragmentation, dysfunction, and motor dysfunction are also central features of other inherited neurodegenerative syndromes, such as amyotrophic lateral sclerosis, Huntington's disease, Parkinson's disease, and Alzheimer's disease. Because there are no treatments that directly enhance mitochondrial fusion or transport, these diseases are considered inexorable and irreversible.

[0087] Examples of mitochondrial-related diseases, disorders, and conditions include, but are not limited to, Alzheimer's disease, Parkinson's disease, Huntington's disease, Charcot-Marie-Tooth disease (type 2A) (CMT), hereditary motor and sensory neuropathies, autism, ADOA, muscular dystrophy, Lou Gehrig's disease, cancer, mitochondrial myopathy, DAD, LHON, Leigh's syndrome, subacute sclerosing encephalopathy, NARP, MNGIE, MERRF, MELAS, mtDNA depletion, MNGIE, autonomic neuropathic mitochondrial myopathy, mitochondrial channelopathies, and / or PDCD / PDH.

[0088] Conditions that may be treated with the methods described herein include, but are not limited to, slow growth, loss of muscle coordination, muscle paralysis and muscle atrophy, vision problems, hearing problems, learning disabilities, heart disease, liver disease, kidney disease, gastrointestinal disorders, respiratory disorders, neurological problems, autonomic dysfunction, and dementia.

[0089] Neurodegenerative diseases

[0090] As described herein, trans-6-phenylhexanamide mitofusin activators rapidly reverse mitochondrial dysmotility in sciatic nerve axons in a mouse model of Charcot-Marie-Tooth disease type 2A. Because impaired mitochondrial fusion, health, and / or trafficking contribute to neurodegeneration in various neurodegenerative diseases (e.g., Charcot-Marie-Tooth disease (CMT2A), Huntington's disease, Parkinson's disease, and Alzheimer's disease, particularly ALS), the present disclosure provides compositions (e.g., compositions comprising mitofusin activators) and methods for treating such neurodegenerative diseases, disorders, and / or conditions.

[0091] Examples of neurodegenerative diseases, disorders, and conditions include diseases in which neuronal mitochondrial dynamism or transport is impaired, such as, but not limited to, hereditary motor and sensory neuropathies (HMSN) (e.g., CMT1 (dominantly inherited, hypertrophic, primarily demyelinating), CMT2 (dominantly inherited, primarily axonal), Dejerine-Sottas (severe form with onset in infancy), CMTX (inherited in an X-linked manner), and CMT4 (including various autosomal recessive demyelinating forms of Charcot-Marie-Tooth disease)), hereditary sensory and autonomic neuropathies (HMSN), -IE, hereditary sensory autonomic neuropathy type II, hereditary sensory autonomic neuropathy type V, HMSN types 1A and 1B (e.g., dominantly inherited hypertrophic demyelinating neuropathies), HMSN type 2 (e.g., dominantly inherited neuropathic neuropathies), HMSN type 3 (e.g., hypertrophic neuropathy of infancy [Déjerine-Sottas]), HMSN type 4 (e.g., hypertrophic neuropathy associated with phytanic acid excess [Refsum]), HMSN type 5 (associated with spastic paraplegia) and / or HMSN type 6 (e.g., with optic nerve atrophy).

[0092] Other examples of neurodegenerative diseases, disorders, and conditions include Alzheimer's disease, ALS, Alexander disease, Alpers disease, Alpers-Huttenlocher syndrome, alpha-methylacyl-CoA racemase deficiency, Andermann syndrome, Aerts syndrome, ataxic neuropathy spectrum, ataxia (e.g., with oculomotor apraxia, autosomal dominant cerebellar ataxia, hearing loss, and narcolepsy), autosomal recessive spastic ataxia of Charlevoix-Saguenay, Batten disease, beta-propeller protein-associated neuropathies, and neuropathy. Neurodegeneration, cerebro-oculofacial-skeletal syndrome (COFS), corticobasal degeneration, CLN1 disease, CLN10 disease, CLN2 disease, CLN3 disease, CLN4 disease, CLN6 disease, CLN7 disease, CLN8 disease, cognitive dysfunction, congenital insensitivity to pain and anhidrosis, dementia, familial encephalopathy with neuroserpin inclusions, familial British dementia, familial Danish dementia, fatty acid hydroxylase-related neurodegeneration, Friedreich's ataxia, Gerstmann-Sträussler-Scheinker disease, GM2-gangliosidosis (e.g., AB variant), HMSN7 (e.g., retinitis pigmentosa) with orthostatic hypotension), Huntington's disease, infantile neuroaxonal dystrophy, infantile-onset ascending hereditary spastic paraplegia, infantile-onset spinocerebellar ataxia, juvenile primary lateral sclerosis, Kennedy disease, kuru, Leigh's disease, Marinesco-Sjögren's syndrome, mild cognitive impairment (MCI), mitochondrial membrane protein-associated neurodegeneration, motor neuron disease, single limb muscular atrophy, motor neuron disease (MND), multiple system atrophy, multiple system atrophy with orthostatic hypotension (Shy-Drager syndrome), multiple sclerosis, multiple system atrophy, neurodegeneration in Down's syndrome (NDS), neurodegeneration associated with age These conditions include, but are not limited to, neurodegeneration associated with brain iron accumulation, neuromyelitis optica, pantothenate kinase-associated neurodegeneration, opsoclonus-myoclonus, prion diseases, progressive multifocal leukoencephalopathy, Parkinson's disease, Parkinson's disease-related disorders, lipomembranous polycystic osseous dysplasia with sclerosing leukoencephalopathy, prion diseases, progressive external ophthalmoplegia, riboflavin transporter deficiency neuronal disorder, Sandhoff disease, spinal muscular atrophy (SMA), spinocerebellar ataxia (SCA), striatonigral degeneration, transmissible spongiform encephalopathies (prion diseases), and / or Wallerian degeneration.

[0093] Charcot-Marie-Tooth (CMT) disease type 2A

[0094] Charcot-Marie-Tooth disease type 2A (CMT2A) is an example of an incurable neurodegenerative disease / axonal neuropathy, disorder, or condition caused by mutations in MFN2, for which no disease-modifying treatment currently exists. As described herein, severely impaired mitochondrial transport from neuronal cell bodies in the spinal cord to synapses in distal neurons in the lower limbs or hands (in addition to the widely recognized smaller mitochondrial size) has been discovered to be a central factor in the development and progression of CMT2A disease. CMT2A is a progressive neuromuscular disease that typically causes muscle weakness and wasting in the distal legs / feet, then the upper limbs, in children aged 1 to 8 years, ultimately leading to severe muscle wasting, skeletal deformity, and permanent disability. The present disclosure provides correction of impaired neuronal mitochondrial transport as a therapeutic target for this disease. The data showed that administration of trans-6-phenylhexanamide mitofusin activator promoted mitochondrial movement along neuronal axons in mouse models where mitochondria were not previously moving and reversed disease-associated deficits in neuromuscular function, including any neuropathic condition (e.g., Huntington's disease, ALS, ALS-like sclerosis, and / or Alzheimer's disease).

[0095] Neurological and neurodegenerative diseases

[0096] As described herein, trans-4-hydroxycyclohexyl derivative mitofusin activators rapidly reverse mitochondrial motility dysfunction in sciatic nerve axons of a mouse model of Charcot-Marie-Tooth disease type 2A. Impaired mitochondrial transport is now believed to also contribute to neurodegeneration in various neurological diseases (e.g., Huntington's disease, Parkinson's disease, and Alzheimer's disease, particularly ALS). Accordingly, the present disclosure provides methods and compositions for treating neurological diseases, disorders, or conditions. For example, the neurological disease, disorder, or condition may be abulia, agraphia, alcoholism, dyslexia, alien hand syndrome, Allan Herndon-Dudley syndrome, alternating hemiplegia of childhood, Alzheimer's disease, amaurosis fugax, amnesia, ALS, aneurysm, Angelman syndrome, nosognosia, aphasia, apraxia, arachnoiditis, Arnold-Chiari malformation, somatognosia, Asperger's syndrome, ataxia, attention deficit hyperactivity disorder, at r-16 syndrome, auditory processing disorder, autism spectrum disorder, Behçet's disease, bipolar disorder, Bell's palsy, brachial plexus injury, brain injury, brain tumor, Brody myopathy, Canavan disease, Capgras delusion, carpal tunnel syndrome, causalgia, central pain syndrome, central pontine myelinolysis, centronuclear myopathy, head disorder, cerebral aneurysm, cerebral arteriosclerosis, cerebral atrophy, cerebral autosomal dominant arteriopathy with subcortical infarction and leukoencephalopathy (CADASIL), cerebral Dysplasia-neuropathy-ichthyosis-keratoderma syndrome (CEDNIK syndrome), cerebral gigantism, cerebral palsy, cerebral vasculitis, cervical spinal stenosis, Charcot-Marie-Tooth disease, Chiari malformation, chorea, chronic fatigue syndrome, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic pain, Cockayne syndrome, Coffin-Lowry syndrome, coma, complex regional pain syndrome, compressive neuropathy, congenital bilateral facial nerve palsy, corticobasal neuropathy Nuclear degeneration, cranial arteritis, craniosynostosis, Creutzfeldt-Jakob disease, cumulative trauma disorder, Cushing's syndrome, cyclothymic disorder, cyclic vomiting syndrome (CVS), cytomegalic inclusion body disease (CIBD), cytomegalovirus infection, Dandy-Walker syndrome, Dawson's disease, Domorsia syndrome, Dejerine-Klumpke palsy, Dejerine-Sottas disease, delayed sleep phase syndrome, dementia, dermatomyositis, developmental coordination disorder,Diabetic neuropathy, diffuse sclerosis, diplopia, impaired consciousness, Down syndrome, Dravet syndrome, Duchenne muscular dystrophy, dysarthria, autonomic neuropathy, dyscalculia, dysgraphia, dyskinesia, dyslexia, dystonia, empty sella syndrome, encephalitis, brain herniation, trigeminal nerve region angiomatosis, encopresis, enuresis, epilepsy, female epilepsy - intellectual disability, Erb's palsy, erythromelalgia, essential tremor, exploding head syndrome, Fabry disease, Fahr's syndrome, syncope, familial spastic paralysis, febrile convulsions, Fisher syndrome, Friedreich's ataxia, fibromyalgia, Foville syndrome, fetal alcohol syndrome, fragile X syndrome, fragile X-associated tremor / ataxia syndrome (FXTAS), Gaucher disease, generalized epilepsy febrile seizures plus, Gerstmann's syndrome, giant cell arteritis, giant cell inclusion disease, globoid cell leukodystrophy, heterotopic gray matter, Guillain-Barré syndrome, generalized anxiety disorder, HTLV-1-associated myelopathy, Hallervorden-Spatz syndrome, head injury, headache, hemifacial Face spasm, hereditary spastic paraplegia, hereditary polyneuropathy ataxia, otic varicella, herpes zoster, Hirayama syndrome, Hirschsprung's disease, Holmes-Addie syndrome, holoprosencephaly, Huntington's disease, hydranencephaly, hydrocephalus, hypercortisolism, hypoxia, immune-mediated encephalomyelitis, inclusion body myositis, incontinentia pigmenti, infantile Refsum's disease, infantile spasms, inflammatory myopathy, intracranial cyst, increased intracranial pressure, isodicentric 15, Joubert syndrome, Karak syndrome, Kearns-Sayre syndrome, Kinsbourne syndrome, Kleine-Lewin syndrome, Klippel-Feil syndrome, Krabbe disease, Kufo-Rakeb syndrome, Lafora disease, Eaton-Lambert myasthenic syndrome, Landau-Kleffner syndrome, lateral medullary (Wallenberg) syndrome, learning disability, Leigh disease, Lennox-Gastaut syndrome, Lesch-Nyhan syndrome, leukodystrophy, leukoencephalopathy with vanishing white matter, dementia with Lewy bodies, lissencephaly, locked-in syndrome, Lou Gehrig's disease (amyotrophic lateral sclerosis (ALS)), lumbar discopathy, lumbar spinal stenosis, Lyme disease - neurological sequelae, Machado-Joseph disease (spinocerebellar ataxia type 3), megalencephalopathy, macropsia, post-landing syndrome (mal de debarquement),Macrocephalic leukoencephalopathy with subcortical cysts, megalencephaly, Melkersson-Rosenthal syndrome, Meniere's disease, meningitis, Menkes disease, metachromatic leukodystrophy, microcephaly, micropsia, migraine, Miller-Fisher syndrome, ministroke (transient ischemic attack), sound aversion, mitochondrial myopathy, Moebius syndrome, single limb muscular atrophy, Morvan's syndrome, motor neuron disease - see ALS, motor skills disorder, moyamoya disease, mucopolysaccharidosis, multi-infarct dementia, multifocal motor neuropathy, multiple sclerosis, multiple system atrophy, muscular dystrophy, myalgic encephalomyelitis, severe Myasthenia, myelinoclastic diffuse sclerosis, myoclonic encephalopathy of infancy, myoclonus, muscle disorders, myotubular myopathy, myotonia congenita, narcolepsy, neuro-Behçet's disease, neurofibromatosis, neuroleptic malignant syndrome, neurological manifestations of AIDS, neurological sequelae of lupus, neuromyotonia, neuronal ceroid lipofuscinosis, neuronal migration disorders, neuropathy, neuropathy, Niemann-Pick disease, non-24-hour sleep-wake syndrome, nonverbal learning disorder, O'Sullivan-McLeod syndrome, occult spinal dysraphism, spinal dysraphism sequence, Ohtahara syndrome, olivopontocerebellar atrophy, opsoclonus-myoclonus syndrome, optic neuritis, orthostatic hypotension, otosclerosis, overuse syndrome, recurrent vision, paresthesia, Parkinson's disease, congenital myotonia, paraneoplastic disorders, seizures, Parry-Romberg syndrome, childhood autoimmune neuropsychiatric disorder associated with streptococcus (PANDAS), Pelizaeus-Merzbacher disease, periodic paralysis, peripheral neuropathy, pervasive developmental disorder, phantom limb / phantom pain, photoinduced sneeze reflex, phytanic acid storage disease, Pick's disease, pinched nerve nerve), pituitary tumor, PMG, polyneuropathy, polio, polymicrogyria, polymyositis, porencephaly, post-polio syndrome, postherpetic neuralgia (PHN), postural hypotension, Prader-Willi syndrome, primary lateral sclerosis, prion disease, progressive facial hemimyopathy, progressive multifocal leukoencephalopathy, progressive supranuclear palsy, prosopagnosia, pseudotumor cerebri, quadrant blindness, quadriplegia, rabies, radiculopathy, Ramsay-Hunt syndrome type 1, Ramsay-Hunt syndrome type 2, Ramsay-Hunt syndrome type 3 - see Ramsay-Hunt syndrome, Rasmussen's encephalitis, reflex neurovascular dystrophy, Refsum's disease,REM sleep behavior disorder, repetitive strain injury, restless legs syndrome, retroviral-associated myelopathy, Rett syndrome, Reye syndrome, rhythmic movement disorder, Romberg syndrome, chorea (Saint Vitus'dance, Sandhoff disease, Schilder's disease (two distinct conditions), schizencephaly, sensory processing disorder, septo-optic dysplasia, shaken baby syndrome, herpes zoster, Shy-Drager syndrome, Sjogren's syndrome, sleep apnea, sleeping sickness, snatification, Sotos syndrome, spasticity, spina bifida, spinal cord injury, spinal cord tumor, spinal muscular atrophy, spinal-bulbar muscular atrophy, spinocerebellar ataxia, split encephalopathy, Steele-Richardson-Olszewski syndrome, stiff-person syndrome, stroke, Sturge-Weber syndrome, stuttering, subacute sclerosing panencephalitis, subcortical arteriosclerotic encephalopathy, superficial hemosiderosis, Sydenham's chorea, syncope, synesthesia, syringomyelia, tarsal tunnel syndrome, tardive dyskinesia, tardive dyspnea dysphrenia), Tarlov cyst, Tay-Sachs disease, temporal arteritis, temporal lobe epilepsy, tetanus, tethered spinal cord syndrome, Thomsen's disease, thoracic outlet syndrome, painful tics, Todd's palsy, Tourette's syndrome, toxic encephalopathy, transient ischemic attack, transmissible spongiform encephalopathy, transverse myelitis, traumatic brain injury, tremor, trichotillomania, trigeminal neuralgia, tropical spastic paraparesis, trypanosomiasis, tuberous sclerosis, 22q13 deletion syndrome, Unverricht-Lundborg disease, vestibular schwannoma (acoustic neuroma), von Hippel-Lindau disease (VHL), Willeujsk encephalomyelitis (VE), Wallenberg syndrome, West syndrome, whiplash injury, Williams syndrome, Wilson's disease, Y-linked deafness, and / or Zellweger syndrome.

[0097] Chemotherapy-induced peripheral neuropathy (CIPN)

[0098] Cancer chemotherapy-induced sensorimotor neuropathy can be prevented or treated using the compositions and methods described herein. Cancer remains a leading cause of death worldwide, but early detection and improved cancer chemotherapy drugs that preferentially attack rapidly dividing cells have had a positive impact on this disease. As a result, the number of cancer survivors is increasing, and the adverse side effects of successful cancer treatment are becoming an increasingly significant problem for cancer survivors. Chemotherapy-induced peripheral neuropathy is one of the most common complications of cancer chemotherapy, affecting 20% ​​of all patients and nearly 100% of patients receiving high-dose chemotherapy. Dose-dependent neurotoxicity of motor and sensory neurons can lead to chronic pain, hypersensitivity to heat, cold, and mechanical stimuli, and / or impaired neuromuscular control. The most common chemotherapeutic agents associated with CIPN are platinum, vinca alkaloids, taxanes, epothilones, and the targeted proteasome inhibitor bortezomib.

[0099] CIPN most commonly affects peripheral sensory neurons whose cell bodies are located in the dorsal root ganglion, which lacks the blood-brain barrier that protects other components of the central and peripheral nervous systems. Unprotected dorsal root ganglion neurons are more susceptible to neuronal hyperexcitability and innate immune system activation induced by circulating cytotoxic chemotherapy agents. CIPN can affect quality of life and lead to disability because it induces chronic neuropathic pain that is resistant to analgesic treatment, as in neuralgia of other origins (e.g., postherpetic neuralgia, diabetic mononeuropathy). Motor nerve involvement typically manifests as impaired handwriting, loss of fine motor function with difficulty buttoning or sewing, and occasionally weakness or loss of endurance in the upper and lower limbs. CIPN usually manifests within a few weeks of chemotherapy and often improves after chemotherapy treatment ends, although residual pain, sensory, or motor deficits persist in one-third to one-half of affected patients. Unfortunately, the limited administration of chemotherapy due to CIPN can lead to delays, reductions, or cessation of cancer treatment, thereby shortening survival time.

[0100] Mitochondrial dysfunction and oxidative stress are associated with CIPN because of the ultrastructural morphological abnormalities observed, impaired mitochondrial DNA transcription and replication, induction of the mitochondrial apoptotic pathway, and the reduction of experimental CIPN symptoms by prior mitochondrial protection. As described herein, trans-4-hydroxycyclohexyl derivative mitofusin activators improve overall mitochondrial function in injured neurons, enhance mitochondrial transport to areas of neuronal damage, and accelerate neuronal repair / regeneration in vitro after chemotherapy-induced injury. Therefore, trans-4-hydroxycyclohexyl derivative mitofusin activators are believed to reduce neuronal injury caused by chemotherapy agents and accelerate the regeneration / repair of nerves damaged by chemotherapy-induced anticancer agents in CIPN. Further validation of the CIPN injury prevention / repair and regeneration hypothesis will be developed by evaluating the in vivo efficacy of trans-4-hydroxycyclohexyl derivative mitofusin activators. Accordingly, the present disclosure provides compositions and methods for treating cancer chemotherapy-induced nerve damage and neuropathy.

[0101] CNS or PNS injury or trauma

[0102] Injuries in the CNS or PNS (e.g., trauma to the CNS or PNS, crush injury, SCI, TBI, stroke, optic nerve injury, or related conditions involving axonal transection) can be treated with the compositions and methods described herein. The CNS includes the brain and spinal cord, and the PNS is composed of cranial nerves, spinal nerves, and autonomic nerves that connect to the CNS.

[0103] Injury to the nervous system caused by mechanical, thermal, chemical, or ischemic agents can impair various nervous system functions, such as memory, cognition, language, and voluntary movement. In most cases, this occurs due to accidental crushing or severing of nerve tracts or as an unintended consequence of medical intervention that interrupts normal communication between nerve cell bodies and their targets. Other types of injury may include disruption of the interrelationship between neurons and their supporting cells or disruption of the blood-brain barrier.

[0104] As described herein, trans-4-hydroxycyclohexyl derivative mitofusin activators rapidly reverse mitochondrial motility dysfunction in neurons from mice or patients with various genetic or chemotherapy-induced neurodegenerative diseases, axons damaged by chemotherapy, and axons severed by physical injury. Therefore, enhancement of mitochondrial transport by trans-4-hydroxycyclohexyl derivative mitofusin activators is believed to enhance the regeneration / repair of physically damaged nerves, such as those caused by vehicle and sports injuries, penetrating trauma from military or criminal acts, and iatrogenic injuries during invasive medical procedures. Further testing of the injury-regeneration hypothesis will be conducted using small molecule mitofusin activators to evaluate their in vivo efficacy. Thus, the present disclosure provides compositions and methods for treating physical nerve injuries.

[0105] As disclosed herein, mitochondrial motility is associated with neuropathy. Mitochondrial motility is also believed to be associated with nerve injury due to traumatic crush or transection. After nerve transection or crush injury, nerves either regenerate and restore neuromuscular function, or they do not regenerate, resulting in permanent impairment of neuromuscular function. The trans-4-hydroxycyclohexyl derivative mitofusin activators described herein can increase mitochondrial transport, enabling nerves to regenerate after traumatic injury.

[0106] formulation

[0107] The agents and compositions described herein can be formulated in any conventional manner using one or more pharmaceutically acceptable carriers or excipients, as previously described (e.g., Remington's Pharmaceutical Sciences (AR Gennaro, Ed.), 21st edition, ISBN: 0781746736 (2005), which is incorporated herein by reference for its disclosure of pharmaceutically acceptable carriers). Such formulations contain a therapeutically effective amount of a biologically active agent described herein, which may be in purified form, together with a suitable amount of carrier, thereby providing a form for proper administration to a subject.

[0108] The term "formulation" refers to a preparation of a drug in a form suitable for administration to a subject, such as a human or a companion or livestock animal. Thus, a "formulation" can include pharmaceutically acceptable excipients, including diluents or carriers.

[0109] As used herein, the term "pharmaceutically acceptable" describes a substance or component that does not cause an unacceptable loss of pharmacological activity or unacceptable adverse side effects. Those skilled in the art are familiar with suitable pharmaceutically acceptable substances. Examples of pharmaceutically acceptable ingredients include those having a monograph in the United States Pharmacopeia (USP 29) and National Formulary (NF 24), United States Pharmacopeial Convention, Inc., Rockville, Maryland, 2005 ("USP / NF") or more recent editions, as well as those listed in the FDA's continuously updated Inactive Ingredient Search online database. Other useful ingredients not listed in the USP / NF may also be used.

[0110] As used herein, the term "pharmaceutically acceptable excipients" includes solvents, dispersion media, coatings, antibacterial agents, antifungal agents, isotonic agents, and absorption delaying agents. The use of such media and agents for pharmaceutically active substances is well known in the art (see generally Remington's Pharmaceutical Sciences (AR Gennaro, Ed.), 21st edition, ISBN: 0781746736 (2005)). Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.

[0111] A "stable" formulation or composition refers to a composition that has sufficient stability to permit storage at a convenient temperature, such as from about 0°C to about 60°C, for a commercially reasonable period of time, e.g., at least about 1 day, at least about 1 week, at least about 1 month, at least about 3 months, at least about 6 months, at least about 1 year, or at least about 2 years.

[0112] The formulation should be suitable for the desired mode of administration. The agents useful in the present disclosure can be formulated by known methods for administration to a subject using several routes, including, but not limited to, parenteral, pulmonary, oral, topical, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, ocular, buccal, and rectal. Individual agents can also be administered in combination with one or more additional agents, or with other biologically active or biologically inactive agents. Such biologically active or biologically inactive agents can be in fluid or mechanical communication with the agent, or can be bound to the agent by ionic, covalent, van der Waals, hydrophobic, hydrophilic, or other physical forces.

[0113] Controlled-release (or sustained-release) preparations can be formulated to extend the activity of an active agent and reduce administration frequency. Controlled-release preparations can also be used to affect the time of onset of action or other characteristics, such as blood levels of the agent, and consequently, the occurrence of side effects. Controlled-release preparations can be designed to initially release an amount of the agent that produces the desired therapeutic effect, and gradually and continuously release other amounts of the agent that maintain that level of therapeutic effect over an extended period of time. To maintain a near-constant level of the agent in the body, the agent can be released from its dosage form at a rate that replaces the amount of agent metabolized or excreted from the body. Controlled-release of an agent can be stimulated by various inducers (e.g., changes in pH, temperature, enzymes, water, or other physiological conditions or molecules).

[0114] The agents or compositions described herein can also be used in combination with other treatment modalities, as further described below. Thus, in addition to the treatments described herein, a subject can also be provided with other treatments known to be effective for treating the disease, disorder, or condition.

[0115] Treatment method

[0116] Also provided herein is a process for treating a mitochondrial-associated disease, disorder, or condition in a subject in need thereof, wherein the subject is administered a therapeutically effective amount of a trans-4-hydroxycyclohexyl derivative mitofusin activator to prevent or treat the mitochondrial-associated disease, disorder, or condition.

[0117] For example, the compositions and methods described herein may also be used as a primary therapy for Charcot-Marie-Tooth or an adjunctive therapy for Huntington's disease, Parkinson's disease, Alzheimer's disease, or ALS to slow or reverse disease progression.

[0118] As another example, the compositions and methods described herein can be used to prevent or treat chemotherapy-induced peripheral neuropathy. For example, they can be used as pre- and post-treatment for individuals scheduled to undergo chemotherapy for cancer treatment. Pre- and post-chemotherapy treatment with a trans-4-hydroxycyclohexyl derivative mitofusin activator can prevent, attenuate, and accelerate recovery from chemotherapy-induced peripheral neuropathy. This treatment can minimize the sensitivity of sensory and motor neurons to chemotherapeutic agents and accelerate repair of chemotherapy-induced nerve damage by promoting mitochondrial health and facilitating mitochondrial localization to areas of injury and regrowth.

[0119] As yet another example, the compositions and methods described herein can be used to treat physical injuries, such as as a primary therapy for any contusion or laceration involving the spine or peripheral nerves, such as motor vehicle or sports injuries (and perhaps even the brain, i.e., concussion). This treatment can help restore normal motor function by enhancing the regeneration and repair of damaged neurons.

[0120] The methods described herein are typically performed on a subject in need thereof. A subject in need of the therapeutic methods described herein may have, have been diagnosed with, be suspected of having, or be at risk for developing a mitochondrial-related disease, disorder, or condition. The determination of the need for treatment is typically assessed by medical history and a physical examination consistent with the disease or condition in question. Diagnosis of various conditions treatable by the methods described herein is within the skill of the art. The subject may be an animal subject, including mammals such as horses, cows, dogs, cats, sheep, pigs, mice, rats, monkeys, hamsters, guinea pigs, and chickens, as well as humans. For example, the subject may be a human subject.

[0121] In general, a safe and effective amount of a trans-4-hydroxycyclohexyl derivative mitofusin activator is, for example, an amount that can induce a desired therapeutic effect in a subject while minimizing undesired side effects. In various embodiments, an effective amount of a mitofusin-modulating agent described herein can substantially inhibit a mitochondrial-related disease, disorder, or condition, slow the progression of a mitochondrial-related disease, disorder, or condition, or limit the onset of a mitochondrial-related disease, disorder, or condition. For example, a desired therapeutic effect can be a delay (e.g., a delay over three years) in peripheral neuropathy compared to placebo, as assessed by a slower increase in the corrected combined CMT neuropathy score. As another example, a desired therapeutic effect can be a reversal or lack of progression of peripheral neuropathy compared to placebo, as indicated by a lower or stable corrected combined CMT neuropathy score. As yet another example, a desired therapeutic effect can be a reversal or lack of progression of dysregulated motor function or an increase in the regeneration and repair of damaged neurons.

[0122] According to the methods described herein, administration can be parenteral, pulmonary, oral, topical, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, ocular, buccal, or rectal administration.

[0123] A therapeutically effective amount of a mitofusin-modulating agent, when used in the treatments described herein, can be used in pure form, or in the form of a pharmaceutically acceptable salt, if such form exists, with or without pharmaceutically acceptable excipients. For example, compounds of the present disclosure can be administered in an amount sufficient to treat, reverse, prevent, or slow the progression of a mitochondrial-related disease, disorder, or condition, at a reasonable benefit / risk ratio applicable to any medical treatment.

[0124] The amount of the compositions described herein that can be combined with a pharmaceutically acceptable carrier to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. It will be recognized by those skilled in the art that the unit content of the active ingredient contained in an individual dose of each dosage form need not itself constitute a therapeutically effective amount, as the required therapeutically effective amount can be achieved by administering several individual doses.

[0125] The toxicity and therapeutic efficacy of the compositions described herein are measured using LD 50 (lethal dose for 50% of the population) and ED 50 The LD (the dose therapeutically effective in 50% of the population) can be determined by standard pharmaceutical procedures in cell cultures or experimental animals. The dose ratio between toxic and therapeutic effects is the LD 50 / ED 50 It is commonly understood in the art that the therapeutic index can be expressed as a ratio, with the larger the therapeutic index the more optimal.

[0126] The specific therapeutically effective dose level for any particular subject will depend on factors such as the disorder being treated and the severity of the disorder, the activity of the specific compound used, the particular composition used, the age, weight, general health, sex, and diet of the subject, the time of administration, the route of administration, the rate of excretion of the composition used, the duration of treatment, drugs used in combination with or concomitantly with the specific compound used, and similar factors well known in the medical arts (e.g., Koda-Kimble, et al.; (2004) Applied Therapeutics: The Clinical Use of Drugs, Lippincott Williams & Wilkins, ISBN 0781748453; Winter (2003) Basic Clinical Pharmacokinetics, 4 thThe dosage will depend on a variety of factors, including, for example, (see, for example, "Therapeutic Uses of the Compounds of the Present and Future," ed., Lippincott Williams & Wilkins, ISBN 0781741475; Sharqel (2004) Applied Biopharmaceutics & Pharmacokinetics, McGraw-Hill / Appleton & Lange, ISBN 0071375503). For example, it is well within the skill of the art to start with a dosage of the composition lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. If desired, an effective daily dose may be divided into multiple doses for purposes of administration. Consequently, a single dose of the composition may contain such amounts or fractions thereof that make up the daily dose. However, it will be understood that the total daily usage of the compounds and compositions of the present disclosure will be determined by the attending physician within the scope of sound medical judgment.

[0127] Also, each of the conditions, diseases, disorders, and conditions described herein, as well as others, may benefit from the compositions and methods described herein. Generally, treating a condition, disease, disorder, or condition includes preventing or delaying the appearance of clinical symptoms in a mammal that may be afflicted with or susceptible to the condition, disease, disorder, or condition but that has not yet experienced or exhibited clinical or asymptomatic symptoms thereof. Treatment can also include inhibiting the condition, disease, disorder, or condition (e.g., halting or reducing the onset of the disease or at least one clinical or asymptomatic symptom thereof). Furthermore, treatment can include alleviating the disease (e.g., reversing the condition, disease, disorder, or condition, or at least one clinical or asymptomatic symptom thereof). The benefit to a treated subject may be statistically significant, or at least perceptible to the subject or to a physician.

[0128] Administration of the trans-4-hydroxycyclohexyl derivative mitofusin activator can be a single event or over a period of time. For example, the mitofusin activator can be administered daily, weekly, biweekly, or monthly. For the treatment of acute conditions, the treatment period is usually at least several days. Certain conditions may require treatment to last from several days to several weeks. For example, treatment may last for one, two, or three weeks. For chronic conditions, treatment may last from several weeks to several months or even years.

[0129] Treatment consistent with the methods described herein may precede, be concurrent with, or follow conventional treatment modalities for treating, preventing, or slowing the progression of a mitochondrial-related disease, disorder, or condition.

[0130] A trans-4-hydroxycyclohexyl derivative mitofusin activator can be administered simultaneously or sequentially with another agent, such as an antibiotic, an anti-inflammatory drug, or another neuroregenerative or neurotherapeutic agent. For example, a trans-4-hydroxycyclohexyl derivative mitofusin activator can be administered simultaneously with another agent, such as an antibiotic or an anti-inflammatory drug. Simultaneous administration can be achieved through the administration of separate compositions, each containing one or more of a mitofusin activator, an antibiotic, an anti-inflammatory drug, or another agent. Simultaneous administration can also be achieved through the administration of a single composition containing two or more of a mitofusin activator, an antibiotic, an anti-inflammatory drug, or another agent. A trans-4-hydroxycyclohexyl derivative mitofusin activator can be administered sequentially with an antibiotic, an anti-inflammatory drug, or another agent. For example, a trans-4-hydroxycyclohexyl derivative mitofusin activator can be administered before or after the administration of an antibiotic, an anti-inflammatory drug, or another agent.

[0131] Administration

[0132] The agents and compositions described herein can be administered by a variety of means known in the art according to the methods described herein. The agents and compositions can be used therapeutically as exogenous or endogenous materials. An exogenous agent is an agent that is produced or manufactured outside the body and administered to the body. An endogenous agent is an agent that is produced or manufactured within the body by some type of device (biological or otherwise) for delivery into or to another organ in the body.

[0133] As discussed above, administration can be parenteral, pulmonary, oral, topical, transdermal (e.g., transdermal patch), intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, ocular, buccal, or rectal administration.

[0134] The agents and compositions described herein can be administered by a variety of methods well known in the art. Administration methods may include, for example, oral ingestion, direct injection (e.g., systemic or stereotactic), implantation of cells engineered to secrete a factor of interest, drug-releasing biomaterials, polymeric matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, implantable matrix devices, mini-osmotic pumps, implantable pumps, injectable gels and hydrogels, liposomes, micelles (e.g., up to 30 μm), nanospheres (e.g., less than 1 μm), microspheres (e.g., 1-100 μm), reservoir devices, combinations of any of the above, or other suitable delivery vehicles that provide the desired release profile in various ratios. Other controlled-release delivery methods for agents or compositions are known to those skilled in the art and are within the scope of the present disclosure.

[0135] Delivery systems can include, for example, infusion pumps that can be used to administer agents or compositions in a manner similar to that used to deliver insulin or chemotherapy to specific organs or tumors. Typically, when using such systems, the agent or composition is administered in combination with a biodegradable, biocompatible polymer implant that releases the agent over a controlled period of time at a selected site. Examples of polymeric materials include polyanhydrides, polyorthoesters, polyglycolic acid, polylactic acid, polyethylene vinyl acetate, and copolymers and combinations thereof. Furthermore, controlled-release systems can be placed near the therapeutic target, so that only a fraction of the systemic dose is required.

[0136] Agents can be encapsulated and administered in a variety of carrier delivery systems, including microspheres, hydrogels, polymeric implants, smart polymer carriers, and liposomes (see generally Uchegbu and Schatzlein, eds. (2006) Polymers in Drug Delivery, CR Syst BN-10:0849325331). Carrier-based systems for delivering molecular or biomolecular agents can provide intracellular delivery, tailor the release rate of the biomolecule / agent, increase the proportion of biomolecules that reach the site of action, improve transport of the drug to the site of action, allow co-deposition with other agents or excipients, improve agent stability in vivo, extend the residence time of the agent at the site of action by reducing clearance, reduce non-specific delivery of the agent to non-target tissues, reduce irritation caused by the agent, reduce toxicity due to a high initial dose of the agent, alter the immunogenicity of the agent, reduce dosing frequency, improve the taste of the product, or improve the shelf life of the product.

[0137] kit

[0138] Kits are also provided herein. Such kits may include the agents or compositions described herein and, in certain embodiments, instructions for administration. Such kits may facilitate the practice of the methods described herein. When supplied as a kit, the various components of the composition may be packaged in separate containers and mixed immediately before use. Components include, but are not limited to, MFN1, MFN2, a target activating peptide, or a trans-4-hydroxycyclohexyl derivative mitofusin activator. The individual packaging of such components may, if desired, be provided as a pack or dispenser device that can contain one or more unit dosage forms containing the composition. The pack may, for example, comprise metal or plastic foil, such as a blister pack. Such individual packaging of the components may, in certain cases, allow for long-term storage without loss of activity of the components.

[0139] The kit may also contain reagents (e.g., sterile water or saline) in separate containers to be added to the separately packaged lyophilized active component. For example, a sealed glass ampoule may contain the lyophilized component, and in another ampoule, sterile water, sterile saline, or each of these is packaged under a neutral, non-reactive gas such as nitrogen. The ampoule may be made of any suitable material, such as glass, an organic polymer such as polycarbonate, polystyrene, ceramics, metal, or any other material commonly used to hold reagents. Other examples of suitable containers include bottles that may be made from materials similar to ampoules, and envelopes that may have a foil-lined interior, such as aluminum or an alloy. Other containers include test tubes, vials, flasks, bottles, syringes, etc. Containers may have a sterile access port, such as a bottle with a stopper that can be pierced by a hypodermic needle. Other containers may have two compartments separated by an easily removable membrane, which, when removed, allows the components to be mixed. The removable membrane may be glass, plastic, rubber, etc.

[0140] In certain embodiments, kits may be supplied with instructional materials. The instructions may be printed on paper or other substrate and / or may be supplied as an electronically readable medium, such as a floppy disk, mini CD-ROM, CD-ROM, DVD-ROM, Zip disk, videotape, audiotape, etc. Detailed instructions may not physically accompany the kit. Instead, the user may be directed to an internet website designated by the kit manufacturer or distributor.

[0141] The compositions and methods described herein using molecular biology protocols can follow various standard techniques known in the art (e.g., Sambrook and Russell (2006) Condensed Protocols from Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, ISBN-10:0879697717; Ausubel, et al. (2002) Short Protocols in Molecular Biology, 5th ed., Current Protocols, ISBN-10:0471250929; Sambrook and Russell (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, ISBN-10:0879695773; Elhai, J. and Wolk, CP1988. Methods in Enzymology 167, 747-754; Studier (2005) Protein Analysis. Expr Purif. 41(1), 207-234; Gellissen, ed. (2005) Production of Recombinant Proteins: Novel Microbial and Eukaryotic Expression Systems, Wiley-VCH, ISBN-10: 3527310363; Baneyx (2004) Protein Expression Technologies, Taylor & Francis, ISBN-10: 0954523253).

[0142] The definitions and methods set forth herein are provided to better define the present disclosure and to guide those of ordinary skill in the art in the practice of the present disclosure. Unless otherwise noted, terms are to be understood in accordance with conventional usage by those of ordinary skill in the relevant art.

[0143] In some aspects, numbers used to describe and claim certain aspects of the present disclosure, expressing properties such as quantities of ingredients, molecular weights, reaction conditions, and the like, are understood to be modified in some instances by the term "about." In some features, the term "about" is used to indicate that a value includes the standard deviation from the mean for the device or method being employed to measure that value. In some features, the numerical parameters set forth in the specification and appended claims are approximations that may vary depending on the desired properties sought to be obtained by the particular feature. In some aspects, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some aspects of the present disclosure are approximations, the numerical values ​​set forth in the specific examples are reported as precisely as practicable. The numerical values ​​presented in some aspects of the present disclosure may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements. The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually set forth herein.

[0144] In some embodiments, the terms "a," "an," "the," and similar references used in the context of describing particular embodiments (particularly in the context of certain claims below) can be construed to encompass both the singular and the plural, unless clearly indicated otherwise. In some embodiments, the term "or," as used herein, including the claims, is used to mean "and / or," unless expressly indicated to refer to alternatives only or that the alternatives are mutually exclusive.

[0145] The terms "comprise," "having," and "include" are open-ended linking verbs. Any form or tense of one or more of these verbs, such as "comprises," "comprising," "has," "having," "includes," and "including," is open-ended. For example, any method that "comprises," "having," or "includes" one or more steps is not limited to having only those one or more steps and may include other steps that are not listed. Similarly, any composition or device that "comprises," "having," or "includes" one or more features is not limited to having only those one or more features and may include other features that are not listed.

[0146] All methods described herein can be performed in any suitable order unless otherwise indicated herein or clearly contradicted by context. The use of any and all examples or exemplary language (e.g., "etc.") provided with respect to certain embodiments herein is intended merely to better clarify the disclosure and does not limit the scope of the disclosure as otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the disclosure.

[0147] Groupings of alternative elements, embodiments, aspects, or features of the disclosure disclosed herein are not to be construed as limiting. Each group member may be referred to and claimed individually or in any combination with other members of the group or other elements found herein. For reasons of convenience or patentability, one or more members of a group may be included in or deleted from a group. When any such inclusion or deletion occurs, the specification is deemed to include the group as modified, and thus fulfills all Markush group descriptions used in the appended claims.

[0148] The citation of any reference herein should not be construed as an admission that such reference is prior art to the present disclosure.

[0149] Although the present disclosure has been described in detail, it will be apparent that modifications, variations, and equivalent embodiments, features, or aspects may exist without departing from the scope of the present disclosure as defined in the appended claims. Furthermore, it should be recognized that all examples in this disclosure are provided as non-limiting examples. [Example]

[0150] The following non-limiting examples are provided to further illustrate the present disclosure.

[0151] It should be appreciated by those skilled in the art that the procedures disclosed in the examples that follow are approaches found by the inventors to work well in the practice of the present disclosure and can therefore be considered to constitute exemplary modes for its practice. However, those skilled in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific features disclosed and still achieve the same or similar results without departing from the spirit and scope of the present disclosure.

[0152] Example 1. N-(cis-4-hydroxycyclohexyl)-6-phenylhexanamide (MiM111 cis) [ka] To a solution of 6-phenylhexanoic acid 1.2 (200 mg, 1.04 mmol, 196 μL, 1.00 eq.) and cis 4-hydroxycyclohexylamine 1.1 (174 mg, 1.14 mmol, 1.10 eq.) and DIEA (269 mg, 2.08 mmol, 362 μL, 2.00 eq.) in DMF (3 mL) was added HOBt (169 mg, 1.25 mmol, 1.20 eq.) and EDCI (299 mg, 1.56 mmol, 1.50 eq.). The mixture was stirred at 10 °C for 10 h. The mixture was diluted with water (20 mL) and extracted with EtOAc (10 mL × 3). The organic phase was washed with 1 M aqueous HCl (30 mL), dried over Na SO , filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC. N-(cis-4-hydroxycyclohexyl)-6-phenylhexanamide Example 1 (46.19 mg, 0.159 mmol, 15.3% yield) was obtained as a white solid. MS: m / z=290.1 ​​(M+H) + ; 1 H NMR(400MHz):MeOD δ 7.25-7.21(m,2H),7.16-7.11(m,3H),3.59-3.48(m,1H),2.60(t,J=7.6Hz,2H),2.13(t,J =7.6Hz,2H),2.11-1.92(m,2H),1.92-1.83(m,2H),1.65-1.60(m,4H),1.34-1.19(m,6H). 13 C NMR (400MHz):MeOD δ:175.660,143.882,129.590,129.441,126.819,70.591,37.231,36.864,34.965,32.519,31.655,29.804,27.098.

[0153] Example 2. N-(trans-4-hydroxycyclohexyl)-6-phenylhexanamide (MiM111 trans) [ka] To a solution of 6-phenylhexanoic acid 1.2 (200 mg, 1.04 mmol, 196 μL, 1.00 eq.), trans 4-hydroxycyclohexylamine 2.1 (174 mg, 1.14 mmol, 1.10 eq.), and DIEA (269 mg, 2.08 mmol, 362 μL, 2.00 eq.) in DMF (3 mL) was added HOBt (169 mg, 1.25 mmol, 1.20 eq.) and EDCI (299 mg, 1.56 mmol, 1.50 eq.). The mixture was stirred at 10 °C for 10 h. The mixture was diluted with water (20 mL) and extracted with EtOAc (10 mL × 3). The organic phase was washed with 1 M aqueous HCl (30 mL), dried over Na SO , filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC. N-(trans-4-hydroxycyclohexyl)-6-phenylhexanamide Example 2 (46.19 mg, 0.159 mmol, 15.3% yield) was obtained as a white solid. MS: m / z=290.1 ​​(M+H) + ;1H NMR(400MHz):MeOD δ 7.25-7.21(m,2H),7.16-7.11(m,3H),3.59-3.48(m,1H),2.60(t,J=7.6Hz,2H),2.13(t,J =7.6Hz,2H),2.11-1.92(m,2H),1.92-1.83(m,2H),1.65-1.60(m,4H),1.34-1.19(m,6H). 13 C NMR (400MHz):MeOD δ:175.660,143.882,129.590,129.441,126.819,70.591,37.231,36.864,34.965,32.519,31.655,29.804,27.098.

[0154] Example 3: The trans-stereoisomer of MiM111 is an allosteric mitofusin agonist, whereas the cis-stereoisomer of MiM111 is inactive. Previously, chemical synthesis of potential mitofusin activators secured starting materials as mixtures of stereoisomers, resulting in products that were mixtures of two or more isomers. It is recognized that target recognition and biological activity can differ between stereoisomers (Sunden, H., et al.; ChemMedChem 8:1283-94, 2013; Jafurulla, M., et al.; Biochim Biophys Acta 1838:158-63, 2014). However, due to the novelty of small molecule mitofusin activators as a drug class, there is a lack of data or even computer models that can predict whether stereoisomers affect mitofusin activator efficacy and, if so, which stereoisomers have superior properties. Therefore, we separately synthesized, validated, and characterized the cis- and trans-diastereomers of N-(4-hydroxycyclohexyl)-6-phenylhexanamide, designated cis- and trans-MiM111 (Figures 3A-3B). Remarkably, the cis-form did not exhibit detectable mitofusin-activating activity, as measured by mitochondrial elongation, in MFN1- or MFN2-deficient mouse embryonic fibroblasts (Figures 2A-2B). In contrast, the trans-MiM111 was equipotent with the prototypic mitofusin agonist of chimera C, a chemical class described by Rocha, et al. (Science 2018). The functional activities of cis- and trans-N-(4-hydroxycyclohexyl)-6-phenylhexanamides reflect their ability to induce characteristic conformational changes in MFN2 underlying their activation (Fig. 2C), mechanistically linking isomeric structure-dependent biological effects with target association. The cis- and trans-stereoisomers of MiM111 had similar in vitro pharmacokinetic profiles (Table 1). [Table 1]

[0155] Example 4: Trans-stereoisomers of MiM111 bearing oxy-substituted linkers are potent mitofusin activators with a spectrum of passive membrane permeabilization properties. A full series of oxy-substituted linker analogs of trans-N-(4-hydroxycyclohexyl)-6-phenylhexanamide was synthesized and evaluated for fusion activity and pharmacokinetic properties. All oxy-substituted trans-analogues retained excellent fusion activity, but the position of the oxygen within the linker altered the passive membrane permeabilization properties of these compounds. The carbamates exhibited 10-fold higher PAMPA activity compared to the parent trans-N-(4-hydroxycyclohexyl)-6-phenylhexanamide, but exhibited poor microsomal stability (Table 2). [Table 2]

[0156] Example 5: In vivo pharmacological profiling demonstrated that trans-MiM111 is a clinical candidate for CMT2A. trans-MiM111 exhibited high microsomal stability and passive permeability (PAMPA Pe) and a low efflux ratio in NIH MDR1 cells, which was unchanged in the presence of the P-gp inhibitor GF120918 (Table 3), indicating that it is not a P-gp substrate. In this context, it was expected that microsomal stability could be correlated with in vivo plasma t1 / 2 and passive permeability at the CNS level. [Table 3]

[0157] This idea was investigated by in vivo PK studies in mice, the only species for which a preclinical model of human CMT2A is published (Cartoni, R., et al.; Brain 133:1460-9, 2010; Bannerman, P., et al.; PLoS ONE 11:e0167573, 2016; Zhou, Y., et al.; J Clin Invest 130:1756-1771, 2019). 50 A target therapeutic brain level of 30 ng / g (approximately 100 nM) was established, which is 10 times higher than the target therapeutic level of 30 ng / g (approximately 100 nM).

[0158] Comparative mouse plasma and brain levels were measured over time after a single 10 mg / kg intravenous (IV) dose. The Vdss (steady-state volume of distribution) of trans-MiM111 was 0.35 L / kg, and the peak brain level was 2,793 ng / g, with therapeutic levels (>30 ng / g) maintained for more than 2 hours after the single IV dose (Figure 4A).

[0159] Due to its low Vdss, high Pe, and low efflux in NIH MDR1 cells, we speculated that trans-MiM111 might accumulate in the brain over a long period of time. If this were the case, brain levels after a single dose could be misleading regarding therapeutic efficacy. To test this notion, we used osmotic minipumps to subcutaneously (SQ) deliver trans-MiM111 at a daily dose of 60 mg / kg / day for 3 days to achieve steady state and define its elimination kinetics after minipump removal (Figure 4B). The plasma half-life of trans-MiM111 was similar after bolus IV administration and chronic SQ administration (1.1 and 1.33 h, respectively), whereas the brain half-life after chronic infusion was substantially longer (3.37 h vs. 1.06 h after IV). Since trans-MiM111 was 96.7% bound to plasma proteins in mice, the unbound fraction (fu) of trans-MiM111 was 0.033. Plasma [Cpd15B] 総 Plasma [Cpd15B] 遊離 / 未結合Using this factor to convert to a , the calculated ratio of brain levels to free plasma levels after chronic infusion is 10.8.

[0160] Example 6: Trans-MiM111 associates with mitochondrial targets in vivo in a preclinical mouse model of CMT2A. Collectively, the above results demonstrate that trans-MiM111 possesses properties that enable it to activate mitochondrial mitofusin in neurons in CMT2A mice in vivo. In the only published study, a prototypic mitofusin activator, chimeric BA / I, enhanced mitochondrial motility when applied locally to sciatic nerve neurons ex vivo (Rocha, AG, et al.; Science 360:336-41, 2018). Because this chemical class of mitofusin agonists has a very short in vivo plasma half-life of approximately 0.2 hours, the in vivo effects of mitofusin activators on mitochondria in CMT2A neurons were not evaluated. Because trans-MiM111 has an oral bioavailability of >75% (Figure 5A), its ability to associate with mitochondrial targets in peripheral nerves was assessed in vivo after a single oral administration. Transgenic mice expressing the human MFN2 T105M mutation in motor neurons (Rocha AG, et al.; Science 360:336-41, 2018) were administered 50 mg / kg of trans-MiM111, and 6 hours later, blinded investigators assessed mitochondrial motility in sciatic nerve neurons. Trans-MiM111 significantly increased both the number and velocity of motile mitochondria in neurons of CMT2A mice (Figure 5B).

[0161] Example 7: Off-target, specificity, and safety studies demonstrate that trans-MiM111 possesses advanced clinical lead properties for CMT2A. Off-target, specificity, and safety studies (Table 4) conducted to qualify trans-MiM111 for potential clinical translation demonstrated that it is a potent and selective mitofusin activator with a favorable drug profile. It exhibited sub-10 nM potency against both MFN1 and MFN2, while simultaneously exhibiting very low inhibitory activity against cytochrome P450 enzymes, indicating a low potential for drug-drug interactions. Activity screening against hERG, hNAV1.5, hKCNQ, and a panel of 42 receptors / kinases revealed only modest inhibition of dopamine aminotransferase (DAT) and monoamine oxidase (MAO-A; approximately 30% inhibition at 10 μM), indicating a 1,000-fold safety window relative to on-target efficacy and correspondingly limited potential for off-target side effects. [Table 4]

[0162] Materials and Methods

[0163] cell line

[0164] Wild-type MEFs were prepared from E10.5 C57 / bl6 mouse embryos. SV-40 T antigen-immortalized MFN1-null MEFs (CRL-2992), MFN2-null MEFs (CRL-2993), and MFN1 / MFN2 double-null MEFs (CRL-2994) were purchased from ATCC. MEFs were subcultured in DMEM (4.5 g / L glucose) plus 10% fetal bovine serum, 1x essential amino acids, 2 mM L-glutamine, 100 units / mL penicillin, and 100 μg / mL streptomycin.

[0165] Confocal live-cell studies of mitochondria

[0166] Live-cell imaging was performed on an Olympus Diaphot 200 fluorescence microscope equipped with a 60x water-immersion objective. All live cells were grown on 12-well plates with coated glass bottoms and examined in modified Krebs-Henseleit buffer (138 mM NaCl, 3.7 mM KCl, 1.2 mM KH2PO4, 15 mM, 20 mM HEPES, and 1 mM CaCl2) at room temperature.

[0167] Cells were excited using 408 nm (Hoechst), 561 nm (MitoTracker Green and Calcein AM, GFP), or 637 nm (TMRE, MitoTracker Orange, ethidium homodimer-1, and AF594-dextran) laser diodes. For mitochondrial elongation studies, the mitochondrial aspect ratio (long axis / short axis) was calculated using automated edge detection and Image J software. Mitochondrial depolarization was calculated as the percentage of green mitochondria visualized on the merged image of MitoTracker Green and TMRE, expressed as green / (green + yellow mitochondria) × 100.

[0168] Chemical synthesis, purification, and characterization of a novel class of small molecule mitofusin activators

[0169] A new series of stereoisomers of mitofusin activator MiM111 analogs was synthesized de novo. The chemical synthesis of cis- and trans-MiM111 is described here.

[0170] Purification method

[0171] General procedure for preparing cis- and trans-MiM111 (see Examples 1 and 2) [ka]

[0172] To a solution of compound 1 (9.00 g, 41.8 mmol, 1.00 eq.) in ethyl acetate (18.0 mL) was added HCl / dioxane (4 M, 36.0 mL, 3.44 eq.). The mixture was stirred at 20° C. for 1 h. Thin layer chromatography (TLC, petroleum ether / ethyl acetate=1 / 2) confirmed that compound 1 (R f =0.50) was completely consumed and a new main spot (R f =0.02) was formed. The mixture was filtered, and the resulting filter cake was washed with ethyl acetate (10.0 mL × 3), filtered, and concentrated under reduced pressure to give a residue. The residue was used in the next step without purification. Compound 2 (6.08 g, 40.1 mmol, 95.9% yield, HCl salt) was obtained as an off-white solid. [ka]

[0173] Compound 2a (6.92 g, 36.0 mmol, 6.78 mL, 1.00 eq.), HOBt (5.83 g, 43.2 mmol, 1.20 eq.), and EDCI (10.3 g, 54.0 mmol, 1.50 eq.) were added to a solution of compound 2 (6.00 g, 39.6 mmol, 1.10 eq., HCl salt) and DIEA (14.0 g, 108 mmol, 18.8 mL, 3.00 eq.) in dimethylformamide (DMF, 60.0 mL). The mixture was stirred at 25 °C for 16 h. Liquid chromatography with mass spectrometry detection (LCMS, EW18054-2-P1A3) confirmed that compound 2a was completely consumed and the desired MS (R t= 0.684 min, 0.709 min) were detected. The reaction mixture was diluted with ethyl acetate (300 mL) and washed with saturated brine (150 mL × 5). The combined organic layers were washed with 1 N HCl (48.0 mL), dried over anhydrous NaSO, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by prep-HPLC (column: Phenomenex LUNA® C18 250 × 80 mm × 10 μm; mobile phase: [water (10 mM NH4HCO3)-ACN (acetonitrile)]; B%: 38% ACN-68% ACN, 8.5 min) to obtain a residue. The residue was triturated with a 5:1 mixture of petroleum ether:ethyl acetate (240 mL) at 25 °C for 5 minutes to obtain MiM111 (3.53 g, 12.0 mmol, 33.9% yield, 99.93% purity) as an off-white solid.

[0174] HPLC:EW18054-2-P1A, Product:R t = 10.792 min, purity: 99.93% at 210 nm.

[0175] Preparative HPLC

[0176] Purification was carried out using HPLC (HO-MeOH; Agilent 1260 Infinity system equipped with DAD and mass detector. Waters SunFire C18 OBD Prep column, 100Å, 5μm, 19mm x 100mm, equipped with SunFire C18 Prep guard cartridge, 100Å, 10μm, 19mm x 10mm). The material was dissolved in 0.7mL of DMSO. Flow rate: 30mL / min. The purity of the obtained fractions was confirmed by analytical LCMS. When each fraction was obtained in the form of a solution immediately after chromatography, a spectrum was recorded for each fraction. The solvent was evaporated in a stream of N2 at 80°C. Based on the LCMS analysis after chromatography, the fractions were combined. The solid fraction was dissolved in 0.5mL of MeOH and transferred to a pre-weighed and marked vial. The obtained solution was again evaporated in a stream of N2 at 80°C. After drying, the product was identified by LCMS, 1 H NMR and13 It was characterized by C NMR.

[0177] Analysis method

[0178] HPLC / HRMS (ESI )

[0179] LC / MS analysis was performed using an Agilent 1100 Series LC / MSD system equipped with a DAD / ELSD and an Agilent LC / MSD VL (G1956A) or SL (G1956B) mass spectrometer, or an Agilent 1200 Series LC / MSD system equipped with a DAD / ELSD and an Agilent LC / MSD SL (G6130A) or SL (G6140A) mass spectrometer. All LC / MS data were acquired using positive / negative mode switching. Compounds were separated using Zorbax SB-C18 1.8 μm 4.6 × 15 mm Rapid Resolution cartridges (PN821975-932) in the following mobile phases: A - ACN, 0.1% formic acid; B - water (0.1% formic acid). Flow rate: 3 mL / min; gradient: 0 min - 100% B; 0.01 min - 100% B; 1.5 min - 0% B; 1.8 min - 0% B; 1.81 min - 100% B; injection volume: 1 μL; ionization mode: atmospheric pressure chemical ionization (APCI), scan range: m / z 80–1000.

[0180] statistical methods

[0181] Time course and dose-response data were calculated for each study using GraphPad Prism. All data are reported as mean ± SEM. Statistical comparisons (two-tailed) were performed using one-way analysis of variance and Tukey's test for multiple groups or Student's t-test for paired comparisons. p<0.05 was considered significant. In vitro pharmacokinetic analysis of mitofusin activators was performed at WuXi Apptec Co. Ltd.

[0182] Binding to human and CD-1 mouse plasma proteins was measured using equilibrium dialysis. Pooled individual frozen EDTA-anticoagulated mouse and human plasma samples were used as test matrices. Warfarin was used as a positive control. Test compounds were added to blank matrix at a final concentration of 2 μM. A 150 μL aliquot of the matrix sample was added to one side of a chamber in a 96-well equilibrium dialysis plate (HTD dialysis), and an equal volume of dialysis buffer was added to the other side. An aliquot of the matrix sample was collected before incubation and used as the TO sample for calculating recovery. Incubations were performed in triplicate. The dialysis plate was placed in a humidified incubator and slowly rotated at 37°C for 4 hours. After incubation, samples were collected from the matrix side and the buffer side. Plasma samples were matched with an equal volume of blank buffer, and buffer samples were matched with an equal volume of blank plasma. Matrix-matched samples were quenched with a stop solution containing an internal standard. All samples were analyzed by LC-MS / MS. Concentrations of all test compounds in matrix and buffer samples are expressed as analyte / internal standard peak area ratios (PAR).

[0183] In vitro stability was measured in human and mouse liver microsomes. An intermediate solution (100 μM small molecule) was first prepared in methanol and subsequently used to prepare the working solution. This was achieved by a 10-fold dilution step of the intermediate solution in 100 mM potassium phosphate buffer. Ten microliters of compound working solution or control working solution was added to all wells of the 96-well plate at that time point (minutes) except for the matrix blank: T0, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15, T16, T17, T18, T19, T20, T21, T22, T23, T24, T25, T26, T27, T28, T29, T30, T31, T32, T33, T34, T35, T46, T47, T48, T49, T50, T51, T52, T53, T54, T55, T56, T57, T58, T59, T60, T61, T62, T63, T64, T65, T66, T67, T68, T69, T70, T71, T72, T73, T74, T75, T76, T77, T78, T79, T79, T80, T81, T82, T83, T84, T85, T86, T87, T88, T89, T91, T92, T93, T94, T95, T96, T97, T98, T99, T99, T91, T91, T92, T93, T94, T95, T96, T97, T98, T99, T99, T99, T99, T100, T10 10 , T 20 , T 30 , T 60, NCF60. Microsome solution (680 μL / well) (#452117, Corning; Woburn, MA, USA; #R1000, Xenotech; Kansas City, Kansas, USA; and #M1000, Xenotech; Kansas City, Kansas, USA) was dispensed into a 96-well plate as reservoirs according to the plate map. 80 μL / well was then added to each plate using an ADDA (Apricot Design Dual Arm, Apricot Designs, Inc., Covina, CA, USA), and the mixture of microsome solution and compound was incubated at 37°C for approximately 10 minutes. Next, 10 μL of 100 mM potassium phosphate buffer solution / well was added to NCF60 and incubated at 37°C (a 1-hour timer was started). NADPH (#00616, Sigma-Aldrich, St. Louis, Missouri, USA) regenerating system was pre-warmed and then dispensed into a 96-well plate as a reservoir according to the plate map. Then, 10 μL / well was added to each plate by ADDA to initiate the reaction. To terminate the reaction, 300 μL / well of stop solution (pre-cooled at 4°C, containing 100 ng / mL tolbutamide and 100 ng / mL labetalol as internal standards) was added, and the sampling plate was agitated for approximately 10 minutes. Next, the samples were centrifuged at 4000 rpm for 20 minutes at 4°C. The supernatant was analyzed by LC-MS / MS.

[0184] Parallel artificial membrane permeability assay (PAMPA)

[0185] A 10 μM solution of the small molecule in 5% DMSO (150 μL) was added to each well of the donor plate. The PVDF membrane of the donor plate was pre-coated with 5 μL of a 1% porcine brain polar lipid extract / dodecane mixture. 300 μL of PBS was then added to each well of the PTFE acceptor plate. The donor and acceptor plates were combined and incubated at room temperature for 4 hours with shaking at 300 rpm. To prepare the TO samples, 20 μL of the donor solution was transferred to a new well, followed by the addition of 250 μL of PBS (DF:13.5) and 130 μL of ACN (containing the internal standard). To prepare the acceptor samples, the plate was removed from the incubator, and 270 μL of solution was transferred from each acceptor well and mixed with 130 μL of ACN (containing the internal standard) to form the acceptor samples. To prepare donor samples, 20 μL of solution was transferred from each donor well and mixed with 250 μL of PBS (DF:13.5) and 130 μL of ACN (containing the internal standard) to form the donor sample. Acceptor and donor samples were analyzed by LC-MS / MS.

[0186] The present invention also relates to the following clauses:

[0187] Clause 1. A method for treating a genetic disorder, physical injury, and / or chemical injury of the peripheral nervous system (PNS) or central nervous system (CNS), the method comprising administering to a subject a therapeutically effective amount of a composition comprising one or more trans-stereoisomer 6-phenylhexanamide derivative mitofusin activators or pharmaceutically acceptable salts thereof, wherein the trans-stereoisomer 6-phenylhexanamide derivative mitofusin activators stimulate mitochondrial fusion, promote mitochondrial health, and enhance mitochondrial intracellular transport.

[0188] Clause 2. The composition comprises one or more mitofusin activators, wherein the mitofusin activators have the formula: [ka] or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, In the formula, R 1 is unsubstituted, monosubstituted or polysubstituted C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 Aryl or C 3-8 The method according to clause 1, wherein the aryl group is heterocyclyl.

[0189] Clause 3. The mitofusin activator has the formula: [ka] wherein R 1 teeth, [ka] 3. The method according to any of clauses 1 or 2, wherein

[0190] Article 4.R 1 independently and optionally, acetamide, C 1-8 Alkoxy, amino, azo, Br, C 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 R is substituted with one or more of heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfur dioxide, and / or thiophene; 1 However, if necessary, one or more of acetamide, alkoxy, amino, azo, Br, C 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8substituted with heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfur dioxide and / or thiophene, wherein one or more of the alkyl, cycloalkyl, heteroaryl, heterocyclyl, indole or phenyl substituents optionally further contain the following substituents: acetamido, alkoxy, amino, azo, Br, C 1-8 Alkyl, carbonyl, carboxyl, Cl, cyano, C 3-8 Cycloalkyl, C 3-8 Heteroaryl, C 3-8 4. The method of any of clauses 1 to 3, wherein the compound is substituted with one or more of heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfur dioxide, and thiophene.

[0191] Clause 5. The mitofusin activator [ka] N-(trans-4-hydroxycyclohexyl)-6-phenylhexanamide, [ka] N-(trans-4-hydroxycyclohexyl)-6-phenylhexanamide, [ka] N-(trans-4-hydroxycyclohexyl)-5-phenoxypentanamide, [ka] 4-(benzyloxy)-N-(trans-4-hydroxycyclohexyl)-4-butanamide, [ka] N-(trans-4-hydroxycyclohexyl)-3-phenethoxypropanamide, [ka] N-(trans-4-hydroxycyclohexyl)-2-(3-phenylpropoxy)acetamide, or [ka] 4-Phenylbutyl(trans-4-hydroxycyclohexyl)carbamate 5. The method according to any one of clauses 1 to 4, wherein

[0192] Article 6. Formula II [ka] or a pharmaceutically acceptable salt thereof, wherein Z is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; R 2 and R 3 are independently H, F, alkyl and C 3-7 cycloalkyl, or R 2 and R 3 Together, C 3-7 forming a cycloalkyl or heterocycloalkyl, R 4 and R 5 are independently H, F, alkyl and C 3-7 cycloalkyl, or R 4 and R 5 Together, C 3-7 forming a cycloalkyl or heterocycloalkyl, Y, O, CR 6 R 7 , C.R. 8 =CR 9 , triple bond, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, NR 8 ,S,SO2,SONR 9 , -NR 9 SO2-, -NR 8 CO-, -CONR 8 -or-NR 8 CONR 9 - and R 6 is H, F, alkyl or cycloalkyl, and R 7 is H, F, alkyl or cycloalkyl, or R 6 and R 7 Together, C 3-7 forming a cycloalkyl or heterocycloalkyl, R 8 is H, alkyl or C 3-7 is cycloalkyl, R 9 is H, alkyl or C 3-7 is cycloalkyl, o is 0, 1, 2, 3, 4 or 5; p is 0 or 1; q is 0, 1, 2, 3, 4 or 5, and when o is 1 or more, Y=NR 8 ,S,SO2,SONR 9 , -NR 9 SO2-, -NR 8 CO-, -CONR 8 -, -NR 8 CONR 9 - and the sum of o+p+q is greater than or equal to 3 or less than or equal to 7; A compound of formula II or a pharmaceutically acceptable salt thereof:

[0193] Clause 7. Z is cycloalkyl, heterocycloalkyl, aryl or heteroaryl; Y, O, CR 6 R 7 , cycloalkyl or aryl; R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 is independently selected from H and alkyl; and o is 0, 1, 2, 3, 4, or 5; p is 0 or 1, q is 0, 1, 2, 3, 4 or 5; when o is 1 or greater, X is S or SO; the sum of o+p+q is equal to or greater than 3 or equal to or less than 7, Item 6. A compound according to item 6 or a pharmaceutically acceptable salt thereof.

[0194] Clause 8. Z is aryl or heteroaryl; Y is O, CH2 or cycloalkyl; R 2 , R 3 , R 4 and R 5 is CH2, R 8 is H, alkyl or C 3-7 is cycloalkyl, R 9 H, alkyl and C 3-7 is cycloalkyl, o is 0, 1, 2, 3, 4 or 5; p is 0 or 1, q is 0, 1, 2, 3, 4 or 5; when o is 1 or greater, X is S or SO; the sum of o+p+q is greater than or equal to 3 or less than or equal to 5, A compound according to any one of clauses 6 to 7 or a pharmaceutically acceptable salt thereof.

[0195] Clause 9. Z is aryl or heteroaryl; Y is cyclopropyl or cyclobutyl; R 2 , R 3 , R 4 and R 5 is CH2, R 8 is H, alkyl or C 3-7 is cycloalkyl, R 9 H, alkyl, COR 7 or C 3-7 cycloalkyl or R 8 and R 9 Together, C 3-7 forming a cycloalkyl, o is 0, 1, 2 or 3; p is 1, q is 0, 1, 2 or 3; the sum of o+p+q is greater than or equal to 3 or less than or equal to 5, Item 9. The compound according to any one of items 6 to 8, or a pharmaceutically acceptable salt thereof.

[0196] Clause 10. Z is aryl or heteroaryl; Y is O or CH2; R 2 , R 3 , R 4 and R 5 is CH2, R 8 is H, alkyl or C 3-7 is cycloalkyl, and R 9 H, alkyl, COR 7 or C 3-7 cycloalkyl or R 8 and R 9 Together, C 3-7 forming a cycloalkyl, o is 0, 1, 2, 3 or 4; p is 1, q is 0, 1, 2, 3 or 4; The sum of o+p+q is 5, 10. The compound according to any one of items 6 to 9, or a pharmaceutically acceptable salt thereof.

[0197] Clause 11. Z is aryl or heteroaryl; Y is O or CH2; R 2 , R 3 , R 4 and R 5 is CH2, o is 0, 1, 2, 3 or 4; p is 1, q is 0, 1, 2, 3 or 4; The sum of o+p+q is 5, 11. The compound according to any one of clauses 6 to 10, or a pharmaceutically acceptable salt thereof.

[0198] Clause 12. Z is phenyl or heteroaryl, the heterocyclic moiety containing 1 to 4 atoms independently selected from nitrogen, oxygen and sulfur, and the phenyl or heterocyclic moiety is R 8 , OR 8 , Cl, F, -CN, CF3, -NR 8 R 9 , -SO2NR 8 R 9 , -NR 8 SO2R 9 , -SO2R 9 , -CONR 8 R 10 , -NR 8 COR 10 , C 3-7 has 0 to 4 substituents independently selected from cycloalkyl and heterocycloalkyl; Y is O or CH2; R 2 , R 3 , R 4 and R 5 is CH2, R 8 is H, alkyl or C 3-7 is cycloalkyl, and R 9 H, alkyl, COR 7 or C 3-7 cycloalkyl or R 8 and R 9 Together, C 3-7 forming a cycloalkyl, R 10 are independently alkyl or C 3-7 cycloalkyl; o is 0, 1, 2, 3 or 4; p is 1, q is 0, 1, 2, 3 or 4; The sum of o+p+q is 5, 12. The compound according to any one of clauses 6 to 11, or a pharmaceutically acceptable salt thereof.

[0199] Clause 13. Z is phenyl or heteroaryl, the heterocyclic moiety containing 1 to 3 atoms independently selected from nitrogen, oxygen and sulfur, and the phenyl or heterocyclic moiety is R 8 , OR 8 , Cl, F, -CN, CF3, -NR 8 R 9 , -SO2R 9 , -CONR 8 R 9 , -NR 7 COR 10 , C 3-7 has 0 to 3 substituents independently selected from cycloalkyl and heterocycloalkyl; Y is O or CH2; R 2 , R 3 , R 4 and R 5 is CH2, R 8 is H, alkyl or C 3-7 is cycloalkyl, and R 9 H, alkyl, COR 7 or C 3-7 cycloalkyl or R 8 and R 9 Together, C 3-7 forming a cycloalkyl, R 10 is alkyl or C 3-7 is cycloalkyl, o is 0, 1, 2, 3 or 4; p is 1, q is 0, 1, 2, 3 or 4; The sum of o+p+q is 5, 13. The compound according to any one of clauses 6 to 12, or a pharmaceutically acceptable salt thereof.

[0200] Clause 14. Z is phenyl, 2-pyridinyl, 3-pyridinyl, 4-pyridinyl, 6-pyrimidinyl, 5-pyrimidinyl, 4-pyrimidinyl or 2-pyrimidinyl, and the phenyl, pyridinyl and pyrimidinyl moieties are R 8 , OR 8 , Cl, F, -CN, CF3, -NR8 R 9 , -SO2R 10 , -CONR 8 R 9 and -NR 8 COR 10 and has 0 to 2 substituents independently selected from Y is O or CH2; R 2 , R 3 , R 4 and R 5 is CH2, R 8 is H, alkyl or C 3-7 is cycloalkyl, R 9 H, alkyl, COR 7 or C 3-7 cycloalkyl, or R 8 and R 9 Together, C 3-7 forming a cycloalkyl, R 10 is alkyl or C 3-7 is cycloalkyl, o is 0, 1, 2, 3 or 4; p is 1, q is 0, 1, 2, 3 or 4; The sum of o+p+q is 5, 14. The compound according to any one of clauses 6 to 13, or a pharmaceutically acceptable salt thereof.

[0201] Clause 15. Z is phenyl, 2-pyridinyl, 3-pyridinyl, 4-pyridinyl, 6-pyrimidinyl, 5-pyrimidinyl, 4-pyrimidinyl or 2-pyrimidinyl, and the phenyl, pyridinyl and pyrimidinyl moieties are R 8 , OR 8 , Cl, F, -CN, CF3, -NR 8 R 9 , -SO2R 10 , -CONR 8 R 9 and -NR 8 COR 10 and has 0 to 2 substituents independently selected from Y is O or CH2; R 2 , R 3 , R 4 and R 5 is CH2, R 8 is H, alkyl or C 3-7 is cycloalkyl, and R 9 H, alkyl, COR 7 or C 3-7 cycloalkyl or R 8 and R 9 Together, C 3-7 forming a cycloalkyl, R 10 is alkyl or C 3-7 is cycloalkyl, o is 0, 1, 2, 3 or 4; p is 1, q is 0, 1, 2, 3 or 4; The sum of o+p+q is 5, 15. The compound according to any one of clauses 6 to 14, or a pharmaceutically acceptable salt thereof.

[0202] Article 16. The compound [ka] N-(cis-4-hydroxycyclohexyl)-6-phenylhexanamide, or [ka] N-(trans-4-hydroxycyclohexyl)-6-phenylhexanamide 16. The compound according to any one of clauses 6 to 15, or a pharmaceutically acceptable salt thereof, wherein:

[0203] Article 17. The compound [ka] N-(trans-4-hydroxycyclohexyl)-6-phenylhexanamide, [ka] N-(trans-4-hydroxycyclohexyl)-5-phenoxypentanamide, [ka] 4-(benzyloxy)-N-(trans-4-hydroxycyclohexyl)-4-butanamide, [ka] N-(trans-4-hydroxycyclohexyl)-3-phenethoxypropanamide, [ka] N-(trans-4-hydroxycyclohexyl)-2-(3-phenylpropoxy)acetamide, or [ka] 4-Phenylbutyl(trans-4-hydroxycyclohexyl)carbamate 16. The compound according to any one of clauses 6 to 15, or a pharmaceutically acceptable salt thereof, wherein:

[0204] Clause 18. A method of treating a disease exhibiting a need for a mitofusin activator, the method comprising administering to a mammal in need thereof a therapeutically effective amount of a compound of formula II [ka] or a pharmaceutically acceptable salt thereof, wherein Z is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; R 2 and R 3 are independently H, F, alkyl and C 3-7 cycloalkyl, or R 2 and R 3 Together, C 3-7forming a cycloalkyl or heterocycloalkyl, R 4 and R 5 are independently H, F, alkyl and C 3-7 cycloalkyl, or R 4 and R 5 Together, C 3-7 forming a cycloalkyl or heterocycloalkyl, Y, O, CR 6 R 7 , C.R. 8 =CR 9 , triple bond, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, NR 8 ,S,SO2,SONR 9 , -NR 9 SO2-, -NR 8 CO-, -CONR 8 -or-NR 8 CONR 9 - and R 6 is H, F, alkyl or cycloalkyl, and R 7 is H, F, alkyl or cycloalkyl, or R 6 and R 7 Together, C 3-7 forming a cycloalkyl or heterocycloalkyl, R 8 is H, alkyl or C 3-7 is cycloalkyl, R 9 is H, alkyl or C 3-7 is cycloalkyl, o is 0, 1, 2, 3, 4 or 5; p is 0 or 1; q is 0, 1, 2, 3, 4 or 5, and when o is 1 or more, Y=NR 8 ,S,SO2,SONR 9 , -NR 9 SO2-, -NR 8 CO-, -CONR 8 -, -NR 8 CONR 9- and the sum of o+p+q is greater than or equal to 3 or less than or equal to 7; method.

[0205] Article 19. PNS or CNS disorders: Chronic neurodegenerative conditions in which mitochondrial fusion, health or transport is impaired; Diseases or disorders associated with dysfunction of mitofusin 1 (MFN1) or mitofusin 2 (MFN2), diseases associated with mitochondrial fragmentation, dysfunction or dysmotility; Degenerative neuromuscular conditions such as Charcot-Marie-Tooth disease, amyotrophic lateral sclerosis, Huntington's disease, Alzheimer's disease, and Parkinson's disease, Hereditary motor and sensory neuropathies, autism, autosomal dominant optic atrophy (ADOA), muscular dystrophy, Lou Gehrig's disease, cancer, mitochondrial myopathy, diabetes mellitus with hearing loss (DAD), Leber's hereditary optic neuropathy (LHON), Leigh's syndrome, subacute sclerosing encephalopathy, neuropathy, ataxia, retinitis pigmentosa, and ptosis (NARP), myoneurogastrointestinal encephalopathy (MNGIE), myoclonic epilepsy with ragged-red fibers syndrome (MERRF), mitochondrial myopathy, encephalomyopathy, lactic acidosis, and stroke-like events (MELAS), mtDNA depletion, mitochondrial neurogastrointestinal encephalomyopathy (MNGIE), autonomic mitochondrial myopathy, mitochondrial channelopathy or pyruvate dehydrogenase complex deficiency (PDCD / PDH), diabetic neuropathy, chemotherapy-induced peripheral neuropathy, and / or Crush injury, spinal cord injury (SCI), traumatic brain injury, stroke, optic nerve injury, and related conditions involving axotomy 19. The method of any of clauses 1 to 5 or 18, selected from any one or combination of:

[0206] Clause 20. The mitofusin activator is the following compound: [ka] 1-(3-(5-cyclopropyl-4-phenyl-4H-1,2,4-triazol-3-yl)propyl)-3-(2-methylcyclohexyl)urea (Chimera C), [ka] 1-(2-((5-cyclopropyl-4-phenyl-4H-1,2,4-triazol-3-yl)oxy)ethyl)-3-(2-methylcyclohexyl)urea 20. The method of any of clauses 1 to 5, 18 or 19, with the proviso that the method is not selected from:

[0207] Clause 21. The method of any of clauses 1-5 or 18-20, wherein the composition further comprises a pharmaceutically acceptable excipient.

[0208] Clause 22. A method for treating a genetic or non-genetic neurodegenerative condition, injury, damage or trauma of the CNS or PNS, comprising administering to a subject a therapeutically effective amount of a compound according to any one of clauses 6 to 17.

[0209] Article 23. If the subject: Chronic neurodegenerative conditions in which mitochondrial fusion, health or transport is impaired; Diseases or disorders associated with dysfunction of mitofusin 1 (MFN1) or mitofusin 2 (MFN2), diseases associated with mitochondrial fragmentation, dysfunction or dysmotility; Degenerative neuromuscular conditions such as Charcot-Marie-Tooth disease, amyotrophic lateral sclerosis, Huntington's disease, Alzheimer's disease, and Parkinson's disease, Hereditary motor and sensory neuropathies, autism, autosomal dominant optic atrophy (ADOA), muscular dystrophy, Lou Gehrig's disease, cancer, mitochondrial myopathy, diabetes mellitus with hearing loss (DAD), Leber's hereditary optic neuropathy (LHON), Leigh's syndrome, subacute sclerosing encephalopathy, neuropathy, ataxia, retinitis pigmentosa, and ptosis (NARP), myoneurogastrointestinal encephalopathy (MNGIE), myoclonic epilepsy with ragged-red fibers syndrome (MERRF), mitochondrial myopathy, encephalomyopathy, lactic acidosis, and stroke-like events (MELAS), mtDNA depletion, mitochondrial neurogastrointestinal encephalomyopathy (MNGIE), autonomic mitochondrial myopathy, mitochondrial channelopathy or pyruvate dehydrogenase complex deficiency (PDCD / PDH), diabetic neuropathy, chemotherapy-induced peripheral neuropathy, and / or Crush injuries, spinal cord injuries, traumatic brain injuries, stroke, optic nerve injuries, and related conditions involving axonal transection 23. The method of clause 22, wherein the patient has been diagnosed with or is suspected of having

[0210] Clause 24. A composition comprising a compound according to any one of clauses 6 to 17 and a pharmaceutically acceptable excipient.

[0211] All documents described herein, including any priority documents and / or testing procedures, are incorporated herein by reference for purposes of all jurisdictions where such practice is recognized, to the extent not inconsistent with the present text. While forms of the disclosure have been illustrated and described, as is apparent from the foregoing general description and specific embodiments, various modifications can be made without departing from the spirit and scope of the disclosure. Accordingly, the disclosure is not intended to be limited thereby. For example, the compositions described herein may not include any component or composition not expressly described or disclosed herein. Any method may lack any step not described or disclosed herein. Similarly, the term "comprising" is considered synonymous with the term "including." Whenever the transitional phrase "comprising" appears after a method, composition, element, or group of elements, it is understood that the inventors also contemplate the same composition or group of elements with the transitional phrase "consisting essentially of," "consisting of," "selected from the group consisting of," or "is" after the description of that composition or element, and vice versa. The term "and / or" as used herein in phrases such as "A and / or B" is intended to include "A and B," "A or B," or "A" and "B." Numerical ranges used herein include the numbers recited in the range. For example, a numerical range of "1 wt% to 10 wt%" includes 1 wt% and 10 wt% within the recited range.

[0212] Unless otherwise indicated, all numbers used in the specification and appended claims expressing quantities of ingredients, molecular weights, reaction conditions, and like properties, are understood to be modified in all instances by the term "about." Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and appended claims are approximations that may vary depending upon the desired properties sought to be obtained by the embodiments of the present disclosure. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

[0213] Whenever a numerical range with a lower and upper limit is disclosed, any number and any range falling within that range is also specifically disclosed. Specifically, each range of values ​​disclosed herein (e.g., "about a to about b" or, equivalently, "approximately a to b" or, equivalently, "approximately a to b") is understood to represent all numbers and ranges falling within the broader range of values. Furthermore, terms in the claims have their plain and ordinary meaning unless expressly and unambiguously defined by the patent owner. Furthermore, the indefinite article "a" or "an" used in the claims is defined herein to mean one or more of the element to which it refers.

[0214] One or more exemplary embodiments are presented herein. For clarity, not all features of a physical implementation are described or shown herein. It is understood that the development of a physical embodiment of the present disclosure involves numerous implementation-specific decisions that vary from implementation to implementation and from time to time, such as compliance with system-related, business-related, government-related, and other constraints, to achieve the developer's goals. While the developer's efforts may be time-consuming, such efforts may be routine for one having ordinary skill in the art and the benefit of this disclosure.

[0215] Thus, the present disclosure is well adapted to achieve the ends and advantages stated, as well as those inherent therein. The specific embodiments disclosed above are merely exemplary, as the disclosure may be modified and practiced in different but equivalent manners apparent to those of ordinary skill in the art having the benefit of the teachings herein. Furthermore, no limitations are intended to the details of construction or design shown herein, other than as described in the claims below. It is therefore apparent that the specific exemplary embodiments disclosed above may be changed, combined, or modified, and all such variations are considered within the scope and spirit of the present disclosure. The embodiments illustratively disclosed herein may suitably be practiced in the absence of any element not expressly disclosed herein and / or any optional element disclosed herein.

Claims

1. Formula (II) 【Chemistry 42】 or a pharmaceutically acceptable salt thereof, wherein Z is aryl; Y is O, CH ２ or cycloalkyl, R ２ , R ３ , R ４ and R ５ is H, o is 0, 1, 2, 3, or 4; p is 0 or 1; When q is 0, 1, 2, 3, 4, or 5, and Y is cycloalkyl and p is 1, then the sum of o+p+q is 3, 4, or 5; otherwise, the sum of o+p+q is 5.

2. Formula (II) 【Chemistry 42】 or a pharmaceutically acceptable salt thereof, wherein Z is phenyl, and said phenyl is R ８ , OR ８ , Cl, F, -CN, CF ３ , -NR ８ R ９ , -SO ２ NR ８ R ９ , -NR ８ SO ２ R ９ , -SO ２ R ９ , -CONR ８ R １０ , -NR ８ COR １０ , C ３－７ has 0 to 4 substituents independently selected from cycloalkyl and heterocycloalkyl; R ８ is H, alkyl or C ３－７ is cycloalkyl, and R ９ is H, alkyl or C ３－７ cycloalkyl or R ８ and R ９ Together, C ３－７ forms a cycloalkyl, and R １０ is alkyl or C ３－７ is cycloalkyl, Y is O, CH 2 or cycloalkyl; R 2 , R 3 , R 4 and R 5 are H; o is 0, 1, 2, 3, or 4; p is 0 or 1; When q is 0, 1, 2, 3, 4, or 5, and Y is cycloalkyl and p is 1, then the sum of o+p+q is 3, 4, or 5; otherwise, the sum of o+p+q is 5.

3. 3. A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, Y is cyclopropyl or cyclobutyl; o is 0, p is 1, q is 2, 3, or 4 is.

4. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein the compound is a compound represented by the following formula [Chemical Formula 78]: 【Chemical 78】 wherein Z is phenyl, and said phenyl is R ８ , OR ８ , Cl, F, -CN, CF ３ , -NR ８ R ９ , -SO ２ NR ８ R ９ , -NR ８ SO ２ R ９ , -SO ２ R ９ , -CONR ８ R １０ , -NR ８ COR １０ , C ３－７ has 0 to 4 substituents independently selected from cycloalkyl and heterocycloalkyl; R ８ is H, alkyl or C ３－７ is cycloalkyl, and R ９ is H, alkyl or C ３－７ cycloalkyl or R ８ and R ９ Together, C ３－７ forming a cycloalkyl, R １０ is alkyl or C ３－７ It is cycloalkyl.

5. 10. The compound of claim 1 or a pharmaceutically acceptable salt thereof, The compound is 【Chemical 44】 N-(trans-4-hydroxycyclohexyl)-6-phenylhexanamide is.

6. 10. The compound of claim 1 or a pharmaceutically acceptable salt thereof, The compound is 【Chemistry 45】 N-(trans-4-hydroxycyclohexyl)-6-phenylhexanamide, 【Chemistry 46】 N-(trans-4-hydroxycyclohexyl)-5-phenoxypentanamide, 【Chemistry 47】 4-(benzyloxy)-N-(trans-4-hydroxycyclohexyl)-4-butanamide, 【Chemistry 48】 N-(trans-4-hydroxycyclohexyl)-3-phenethoxypropanamide, 【Chemistry 49】 N-(trans-4-hydroxycyclohexyl)-2-(3-phenylpropoxy)acetamide, or 【Chemistry 50】 4-phenylbutyl(trans-4-hydroxycyclohexyl)carbamate is.

7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

8. 8. The pharmaceutical composition of claim 7 for treating a mitochondrial-related disease, disorder, or condition.

9. 9. The pharmaceutical composition of claim 8, wherein the disease, disorder, or condition is a genetic disorder, physical injury, trauma, and / or chemical injury of the central nervous system (CNS) or peripheral nervous system (PNS).

10. 10. The pharmaceutical composition of claim 9, wherein the CNS or PNS disorder is selected from one or a combination of the following: Chronic neurodegenerative conditions in which mitochondrial fusion, health or transport is impaired; Diseases or disorders associated with dysfunction of Mitofusion 1 (MFN1) or Mitofusion 2 (MFN2), Diseases associated with mitochondrial fragmentation, dysfunction or dysmotility, Degenerative neuromuscular conditions such as Charcot-Marie-Tooth disease, amyotrophic lateral sclerosis, Huntington's disease, Alzheimer's disease, and Parkinson's disease, Hereditary motor and sensory neuropathies, autism, autosomal dominant optic atrophy (ADOA), muscular dystrophy, Lou Gehrig's disease, cancer, mitochondrial myopathy, diabetes mellitus with hearing loss (DAD), Leber's hereditary optic neuropathy (LHON), Leigh's syndrome, subacute sclerosing encephalopathy, neuropathy, ataxia, retinitis pigmentosa, and ptosis (NARP), myoneuronectal gastrointestinal encephalopathy (MNGIE), myoclonic epilepsy with ragged-red fibers syndrome (MERRF), mitochondrial myopathy, encephalomyopathy, lactic acidosis, and stroke-like events (MELAS), mtDNA depletion, mitochondrial neurogastrointestinal encephalomyopathy (MNGIE), autonomic mitochondrial myopathy, mitochondrial channelopathy or pyruvate dehydrogenase complex deficiency (PDCD / PDH), diabetic neuropathy, chemotherapy-induced peripheral neuropathy, and Crush injury, spinal cord injury, traumatic brain injury, stroke, optic nerve injury, and related conditions involving axonal transection.

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