U.S. patent application number 17/245388 was filed with the patent office on 2021-08-19 for trans-4-hydroxycyclohexyl phenyl amide mitofusin activators and methods of use thereof.
This patent application is currently assigned to MITOCHONDRIA EMOTION, INC.. The applicant listed for this patent is MITOCHONDRIA EMOTION, INC.. Invention is credited to Gerald W. DORN, II.
Application Number | 20210251925 17/245388 |
Document ID | / |
Family ID | 1000005553235 |
Filed Date | 2021-08-19 |
United States Patent
Application |
20210251925 |
Kind Code |
A1 |
DORN, II; Gerald W. |
August 19, 2021 |
TRANS-4-HYDROXYCYCLOHEXYL PHENYL AMIDE MITOFUSIN ACTIVATORS AND
METHODS OF USE THEREOF
Abstract
Compounds and compositions including stereoisomers of
6-phenylhexanamide derivative small molecule mitofusin activators
are described. In particular, mitofusin activators comprising
derivatives of (trans-4-hydroxycyclohexyl)-6-phenylhexanamide,
which are useful for treating diseases or disorders associated with
a mitochondria-associated disease, disorder, or condition such as
diseases or disorders associated with mitofusin-1 (MFN1) and/or
mitofusin-2 (MFN2), or mitochondrial dysfunction, are described.
Methods of treatment and pharmaceutical formulations are also
described.
Inventors: |
DORN, II; Gerald W.;
(Hamilton, OH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MITOCHONDRIA EMOTION, INC. |
Hamilton |
OH |
US |
|
|
Assignee: |
MITOCHONDRIA EMOTION, INC.
Hamilton
OH
|
Family ID: |
1000005553235 |
Appl. No.: |
17/245388 |
Filed: |
April 30, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16935557 |
Jul 22, 2020 |
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17245388 |
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PCT/US2020/014784 |
Jan 23, 2020 |
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16935557 |
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62949060 |
Dec 17, 2019 |
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62797513 |
Jan 28, 2019 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/165 20130101;
A61K 31/27 20130101; A61P 25/28 20180101 |
International
Class: |
A61K 31/165 20060101
A61K031/165; A61P 25/28 20060101 A61P025/28; A61K 31/27 20060101
A61K031/27 |
Goverment Interests
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0002] This invention was made with government support under grants
R41NS113642 and R41NS115184 awarded by the National Institutes of
Health. The government has certain rights in the invention.
Claims
1. A method comprising: administering a therapeutically effective
amount of a composition comprising one or more metabolites of
trans-(4-hydroxycyclohexyl)-6-phenylhexanamide or a
pharmaceutically acceptable salt thereof to a subject having or
suspected of having a mitochondria-associated disease, disorder or
condition, the one or more metabolites being active to promote
mitofusin activation.
2. The method of claim 1, wherein the one or more metabolites are
selected from the group consisting of ##STR00037## and any
combination thereof.
3. The method of claim 1, wherein the mitochondria-associated
disease, disorder or condition is a peripheral nervous system (PNS)
or central nervous system (CNS) genetic or non-genetic disorder,
physical damage, and/or chemical injury.
4. The method of claim 3, wherein the PNS or CNS disorder is
selected from any one or a combination of a chronic
neurodegenerative condition wherein mitochondrial fusion, fitness,
or trafficking are impaired; a disease or disorder associated with
mitofusin 1 (MFN1) or mitofusin 2 (MFN2) dysfunction; a disease
associated with mitochondrial fragmentation, dysfunction, or
dysmotility; a degenerative neuromuscular condition such as
Charcot-Marie-Tooth disease, Amyotrophic Lateral Sclerosis,
Huntington's disease, Alzheimer's disease, Parkinson's disease;
hereditary motor and sensory neuropathy, autism, autosomal dominant
optic atrophy (ADOA), muscular dystrophy, Lou Gehrig's disease,
cancer, mitochondrial myopathy, diabetes mellitus and deafness
(DAD), Leber's hereditary optic neuropathy (LHON), Leigh syndrome,
subacute sclerosing encephalopathy, neuropathy, ataxia, retinitis
pigmentosa, and ptosis (NARP), myoneurogenic gastrointestinal
encephalopathy (MNGIE), myoclonic epilepsy with ragged red fibers
(MERRF), mitochondrial myopathy, encephalomyopathy, lactic
acidosis, stroke-like symptoms (MELAS), mtDNA depletion,
mitochondrial neurogastrointestinal encephalomyopathy (MNGIE),
dysautonomic mitochondrial myopathy, mitochondrial channelopathy,
or pyruvate dehydrogenase complex deficiency (PDCD/PDH); diabetic
neuropathy; chemotherapy-induced peripheral neuropathy; crush
injury, spinal cord injury (SCI), traumatic brain injury, stroke,
optic nerve injury, and related conditions that involve axonal
disconnection; and any combination thereof.
5. A method comprising: administering a therapeutically effective
amount of a composition comprising
trans-(4-hydroxycyclohexyl)-6-phenylhexanamide or a
pharmaceutically acceptable salt thereof to a subject having or
suspected of having a mitochondria-associated disease, disorder or
condition; and forming one or more metabolites of the
trans-(4-hydroxycyclohexyl)-6-phenylhexanamide in vivo within the
subject, the one or more metabolites being active to promote
mitofusin activation and being selected from the group consisting
of ##STR00038## and any combination thereof.
6. The method of claim 5, wherein the mitochondria-associated
disease, disorder or condition is a peripheral nervous system (PNS)
or central nervous system (CNS) genetic or non-genetic disorder,
physical damage, and/or chemical injury.
7. The method of claim 6, wherein the PNS or CNS disorder is
selected from any one or a combination of a chronic
neurodegenerative condition wherein mitochondrial fusion, fitness,
or trafficking are impaired; a disease or disorder associated with
mitofusin 1 (MFN1) or mitofusin 2 (MFN2) dysfunction; a disease
associated with mitochondrial fragmentation, dysfunction, or
dysmotility; a degenerative neuromuscular condition such as
Charcot-Marie-Tooth disease, Amyotrophic Lateral Sclerosis,
Huntington's disease, Alzheimer's disease, Parkinson's disease;
hereditary motor and sensory neuropathy, autism, autosomal dominant
optic atrophy (ADOA), muscular dystrophy, Lou Gehrig's disease,
cancer, mitochondrial myopathy, diabetes mellitus and deafness
(DAD), Leber's hereditary optic neuropathy (LHON), Leigh syndrome,
subacute sclerosing encephalopathy, neuropathy, ataxia, retinitis
pigmentosa, and ptosis (NARP), myoneurogenic gastrointestinal
encephalopathy (MNGIE), myoclonic epilepsy with ragged red fibers
(MERRF), mitochondrial myopathy, encephalomyopathy, lactic
acidosis, stroke-like symptoms (MELAS), mtDNA depletion,
mitochondrial neurogastrointestinal encephalomyopathy (MNGIE),
dysautonomic mitochondrial myopathy, mitochondrial channelopathy,
or pyruvate dehydrogenase complex deficiency (PDCD/PDH); diabetic
neuropathy; chemotherapy-induced peripheral neuropathy; crush
injury, spinal cord injury (SCI), traumatic brain injury, stroke,
optic nerve injury, and related conditions that involve axonal
disconnection; and any combination thereof.
8. A composition comprising: one or more metabolites of
trans-(4-hydroxycyclohexyl)-6-phenylhexanamide or a
pharmaceutically acceptable salt thereof, the one or more
metabolites being active to promote mitofusin activation.
9. The composition of claim 8, wherein the one or more metabolites
are selected from the group consisting of ##STR00039## and any
combination thereof.
10. The composition of claim 8, further comprising: a
pharmaceutically acceptable excipient.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a continuation of U.S. patent
application Ser. No. 16/935,557, filed on Jul. 22, 2020, which is a
continuation-in-part of International Patent Application
PCT/US2020/014784, filed on Jan. 23, 2020, which claims the benefit
of priority under 35 U.S.C. .sctn. 119 from U.S. Provisional Patent
Applications 62/797,513, filed on Jan. 28, 2019, and 62/949,060,
filed on Dec. 17, 2019, each of which is incorporated herein by
reference in its entirety.
FIELD OF THE INVENTION
[0003] Among the various aspects of the present disclosure is the
provision of novel active trans-4-hydroxycyclohexyl phenyl amide
stereoisomers of small molecule mitofusin activators and methods of
use thereof.
[0004] The present disclosure generally relates to compositions and
methods for treating genetic or acquired central and peripheral
neuropathies, neurodegenerative diseases, disorders, or
conditions.
SUMMARY
[0005] Among the various aspects of the present disclosure is the
provision of novel functionally active stereoisomers of small
molecule mitofusin activators and methods of use thereof.
[0006] One aspect of the present disclosure provides for methods of
treating neuropathies, neurodegenerative diseases, disorders, or
conditions. In some features, the method comprises administering to
a subject a therapeutically effective amount of a composition
comprising one or more active stereoisomers of mitofusin
activators; the mitofusin activators stimulate mitochondrial fusion
and subcellular mitochondrial transport in neurons, thereby evoking
resistance to neuronal injury, accelerating neuronal repair, and
promoting neuronal regrowth and regeneration.
[0007] Another aspect of the present disclosure provides for a
method of activating mitofusin in a subject in need thereof. In
some features, the method comprises administering to a human
subject or affected animal a composition comprising one or more
active trans-4-hydroxycyclohexyl stereoisomers of mitofusin
activators; the active trans-4-hydroxycyclohexyl stereoisomers of
mitofusin activators stimulate mitochondrial fusion and subcellular
mitochondrial transport in neurons, thereby evoking resistance to
neuronal injury, accelerating neuronal repair, and promoting
neuronal regrowth and regeneration; the subject or affected animal
has a genetic or acquired central or peripheral neuropathy,
neurodegenerative disease, disorder, or condition.
[0008] Another aspect of the present disclosure provides for
methods of preventing, mitigating, reducing, or enhancing recovery
from iatrogenic, traumatic, or collateral nerve damage in a subject
in need thereof. In some features, the method comprises
administering to a subject a composition comprising one or more
active trans-4-hydroxycyclohexyl stereoisomers of mitofusin
activators; the active stereoisomers of mitofusin activators
stimulate mitochondrial fusion and subcellular mitochondrial
transport in neurons, thereby evoking resistance to neuronal
injury, accelerating neuronal repair, and promoting neuronal
regrowth and regeneration; the subject or affected animal has a
genetic or acquired central or peripheral neuropathy,
neurodegenerative disease, disorder, or condition.
[0009] In some aspects, the active trans-4-hydroxycyclohexyl
stereoisomers of mitofusin activators have 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]propanamido}-
-4H,5H,6H-cyclopenta[b]thiophene-3-carboxamide (Cpd B, Rocha
Science 2018).
[0010] In some aspects, the active trans-4-hydroxycyclohexyl
stereoisomers of mitofusin activators have 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]propanamido}-
-4H,5H,6H-cyclopenta[b]thiophene-3-carboxamide (Cpd B, Rocha
Science 2018).
[0011] In some aspects, the active trans-4-hydroxycyclohexyl
stereoisomers of mitofusin activators: target mitofusin-1 (MFN1) or
mitofusin-2 (MFN2); increase mitochondrial elongation by enhancing
mitochondrial fusion; enhance mitochondrial function measured as
inner membrane electrochemical polarization; enhance mitochondrial
transport in nerve axons; correct cell and organ dysfunction caused
by primary or secondary mitochondrial abnormalities; reverse
mitochondrial defects (e.g., dysmorphometry, clustering, loss of
polarization, loss of motility); restore, activate, regulate,
modulate, promote, or enhance the fusion, function, tethering,
transport, trafficking (e.g., axonal mitochondrial trafficking),
mobility, or movement of mitochondria (in, optionally, a nerve or a
neuron); enhance mitochondrial elongation or mitochondrial aspect
ratio; disrupt intramolecular restraints in MFN2; allosterically
activate MFN2; and repair morphological and functional defects in
diseased or damaged neurons with mitochondrialabnormalities.
[0012] In some aspects, the active trans-4-hydroxycyclohexyl
stereoisomer mitofusin activator comprises one or more compounds
having structures represented by formula (I):
##STR00001##
or a pharmaceutically acceptable salt, tautomer, or stereoisomer
thereof wherein, R.sup.1 may be a non-, mono-, or poly-substituted
C.sub.3-8 cycloalkyl, C.sub.3-8 heteroaryl, C.sub.3-8 aryl, or
C.sub.3-8 heterocyclyl.
[0013] In some aspects, the active trans-4-hydroxycyclohexyl
stereoisomer mitofusin activator may be a compound having a
structure represented by formula (I)
##STR00002##
wherein R.sup.1 may be one of the following moieties:
##STR00003##
[0014] In some aspects, R.sup.1 may be optionally substituted by
one or more of: acetamide, C.sub.1-8alkoxy, amino, azo, Br,
C.sub.1-3 alkyl, carbonyl, carboxyl, Cl, cyano,
C.sub.3-8cycloalkyl, C.sub.3-8 heteroaryl, C.sub.3-8 heterocyclyl,
hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide,
sulfone, and/or thiophene; and optionally further substituted with
one or more acetamide, alkoxy, amino, azo, Br, C.sub.1-8 alkyl,
carbonyl, carboxyl, Cl, cyano, C.sub.3-8 cycloalkyl, C.sub.3-8
heteroaryl, C.sub.3-8 heterocyclyl, hydroxyl, F, halo, indole, N,
nitrile, O, phenyl, S, sulfoxide, sulfone, and/orthiophene.
Optionally the aforementioned alkyl, cycloalkyl, heteroaryl,
heterocyclyl, indole, or phenyl may be further substituted with one
or more of the following: acetamide, alkoxy, amino, azo, Br,
C.sub.1-3 alkyl, carbonyl, carboxyl, Cl, cyano, C.sub.3-8
cycloalkyl, C.sub.3-8 heteroaryl, C.sub.3-8 heterocyclyl, hydroxyl,
F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfone,
and/or thiophene.
[0015] In some aspects, formula (I) may be one of the following
moieties:
##STR00004##
[0016] Yet another aspect of the present disclosure provides for a
pharmaceutical composition comprising an active
trans-4-hydroxycyclohexyl stereoisomer mitofusin activator,
optionally in combination with one or more therapeutically
acceptable diluents or carriers.
[0017] In some aspects, the pharmaceutical composition comprises a
pharmaceutically acceptable excipient.
[0018] In some aspects, the pharmaceutical composition comprises
one or more of the following: neuroprotectants, anti-Parkinsonian
drugs, amyloid protein deposition inhibitors, beta amyloid
synthesis inhibitors, antidepressants, anxiolytic drugs,
antipsychotic drugs, anti-amyotrophic lateral sclerosis drugs,
anti-Huntington's drugs, anti-Alzheimer's drugs, anti-epileptic
drugs, and/or steroids.
[0019] Yet another aspect of the present disclosure provides for a
method of treating a mitochondria-associated disease, disorder, or
condition in a subject, the method comprising administering to the
subject a therapeutically effective amount of a mitofusin
activator.
[0020] In some aspects, the subject may be diagnosed with or may be
suspected of having a mitochondria-associated disease, disorder, or
condition.
[0021] In some aspects, the mitochondria-associated disease,
disorder, or condition may be one or more of: 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 that involve axonal disconnection; a chronic
neurodegenerative condition wherein mitochondrial fusion, fitness,
or trafficking are impaired; a disease or disorder associated with
mitofusin 1 (MFN1) or mitofusin 2 (MFN2) or mitochondrial
dysfunction, fragmentation, or fusion; dysfunction in MFN1 or MFN2
unfolding; mitochondria dysfunction caused by mutations; a
degenerative neurological condition, such as Alzheimer's disease,
Parkinson's disease, Charcot-Marie-Tooth disease, or Huntington's
disease; hereditary motor and sensory neuropathy, autism, autosomal
dominant optic atrophy (ADOA), muscular dystrophy, Lou Gehrig's
disease, cancer, mitochondrial myopathy, diabetes mellitus and
deafness (DAD), Leber's hereditary optic neuropathy (LHON), Leigh
syndrome, subacute sclerosing encephalopathy, neuropathy, ataxia,
retinitis pigmentosa, and ptosis (NARP), myoneurogenic
gastrointestinal encephalopathy (MNGIE), myoclonic epilepsy with
ragged red fibers (MERRF), mitochondrial myopathy,
encephalomyopathy, lactic acidosis, stroke-like symptoms (MELAS),
mtDNA depletion, mitochondrial neurogastrointestinal
encephalomyopathy (MNGIE), dysautonomic mitochondrial myopathy,
mitochondrial channelopathy, and/or pyruvate dehydrogenase complex
deficiency (PDCD/PDH).
[0022] Other objects and features will be in part apparent and in
part pointed out hereinafter.
DESCRIPTION OF THE DRAWINGS
[0023] Those of skill in the art will understand that the drawings,
described below are for illustrative purposes only. The drawings
are not intended to limit the scope of the present teachings in any
way.
[0024] FIG. 1A illustrates a structural model of human MFN2 in the
closed/inactive (left) and open/active conformation (right) that
may be promoted by mitofusin activators. FIG. 1B depicts a
pharmacophore modeling of interacting amino acid side chains that
resulted in the prototype mitofusin small molecule agonist (Rocha,
et al.; Science 2018).
[0025] FIGS. 2A-2C depict structure-function relationships of MiM
111 cis- and trans-isostereomers (see e.g., Example 3). FIG. 2A
shows the dose-response curves for cis- (Cpd 15A) and trans-(Cpd
15B) MiM 111 stereoisomers in cells expressing only MFN2. FIG. 2B
shows the dose-response curves for cis-(Cpd 15A) and trans-(Cpd
15B) MiM 111 stereoisomers in cells expressing only MFN1. "Cpd 2"
represents Chimera C, a prototype mitofusin activator for
comparison. FIG. 2C shows the FRET analysis of conformational
switching provoked by mitofusin activators. Open conformation is
more active. "Peptide" is MP1 mitofusin agonist peptide described
in reference Franco Nature 2016 and depicted in FIG. 1A.
[0026] FIGS. 3A-3B depict chemical structures and corresponding NMR
spectra of MiM 111 cis- (FIG. 3A) and trans-(FIG. 3B) isostereomers
(see e.g., Example 3). Black arrows and arrowhead are specific to
cis-isomer; grey arrowheads signify trans-isomer.
[0027] FIGS. 4A-4B depict in vivo (mouse) pharmacokinetic
properties of trans-MiM 111 (Cpd 15B) (see e.g., Example 5). FIG.
4A shows the total plasma and brain concentrations after single
dose IV injections. FIG. 4B shows the steady state elimination
kinetics after 3 days of continuous subcutaneous infusion.
[0028] FIGS. 5A-5B shows the oral bioavailability and in vivo
target engagement of trans-MiM 111 (Cpd 15B) (see e.g., Example 6).
FIG. 5A shows the plasma levels after single dose IV (closed
circles) or oral (open circles) administration. FIG. 5B shows
kymographs demonstrating mitochondrial motility in sciatic nerves
of CMT2A mice 6 hours after oral administration as in A. Motile
mitochondria exhibit horizontal displacement (corresponding group
quantitative data are on the right).
DETAILED DESCRIPTION
[0029] The present disclosure is based, at least in part, on the
discovery that pharmacophore modeling of function-critical
MFN2-derived interacting peptides may produce structurally diverse
small molecule peptidomimetic activators useful to treat
mitochondrial-associated diseases, disorders, and conditions. As
shown herein, the present disclosure provides stereoisomer-specific
chemicals or compositions for regulating mitochondrial function.
These compositions may be useful to correct organelle, cell, and
organ dysfunction caused by primary or secondary mitochondrial
abnormalities that cause or contribute to disease pathology and
dysfunction.
Mitofusin Activators
[0030] The present disclosure provides for a class of
stereoisomer-specific trans-4-hydroxycyclohexyl derivative small
molecules that promote a change in configuration of MFN1 and MFN2
leading to heightened activation. As described herein, a
composition for the treatment of a mitochondria-associated disease,
disorder, or condition may comprise an active
trans-4-hydroxycyclohexyl mitofusin activator, such as a
peptidomimetic (e.g., a small-molecule that mimics the
chemico-structural features of a peptide). A peptidomimetic may be
a chemical peptidomimetic. For example, the peptidomimetic may
mimic a mitofusin-derived mini-peptide. Certain metabolites of the
mitofusin activator may also maintain activity in vivo.
[0031] The present disclosure describes functional activity of
small molecule peptidomimetics requiring mimicry of the endogenous
peptide conformation in 3-dimensional space. Diastereomers of the
same chemical mitofusin activator prefer different 3-dimensional
structures (FIGS. 1A-1B). Thus, cis- and trans-4-hydroxycyclohexyl
derivatives exhibit dramatically different abilities to physically
engage with and functionally activate their mitofusin protein
targets (FIG. 2A-2C).
[0032] As described herein, a new generation of
trans-4-hydroxycyclohexyl derivative peptidomimetic small molecules
has been developed exhibiting stereoisomer-specific functional
activity. These compounds activate mitochondrial fusion by
directing MFN1 and MFN2 to different conformational states. The
prototype small molecule peptidomimetics to target MFN1 or MFN2
(described in Rocha, et al.; Science, 2018) had poor
pharmacokinetic characteristics, making them "undruggable."
Described herein are active stereoisomers of a structurally
distinct class of small molecule mitofusin activators that activate
mitochondrial fusion and subcellular transport, have favorable
pharmacokinetic properties, and may be used to correct
mitochondrial and cellular dysfunction.
Mitofusin Mini-Peptide
[0033] As described herein, a peptide mitofusin activator may be an
MFN2-derived mini-peptide as described in Franco, et al.; Nature
2016.
MFN Activator (Fusion-Promoting) Peptidomimetic
[0034] As described herein, a peptidomimetic may be a MFN activator
(fusion-promoting) peptidomimetic that competes with endogenous
MFN1 or MFN2 HR1-HR2 peptide-peptide interactions as described in
Franco, et al.; Nature 2016 and Rocha, et al.; Science 2018.
[0035] The prototype mitofusin activator, Chimera C according to
the present disclosure, includes the following compound:
##STR00005##
Mitofusin Activators: Structurally Distinct Small Molecules that
Activate MFN1 and/or MFN2
[0036] The small molecule mitofusin activators described herein are
allosteric mitofusin activators designed in part using the
pharmacophore HR1-HR2 peptide-peptide interaction model described
in Rocha, et al.; Science 2018, but which are structurally distinct
and of separate chemical classes from those reported by Rocha. An
activator is a substance that partially or fully activates the
protein to which it binds.
[0037] The mitofusin activator may comprise one or more compounds
represented by formula (I):
##STR00006##
or a pharmaceutically acceptable salt, tautomer, or stereoisomer
thereof, wherein R.sup.1 may be selected from the following
moieties:
##STR00007##
[0038] Optionally, R.sup.1 in formula (I) may be independently
substituted by one or more of the following groups: acetamide,
C.sub.1-8 alkoxy, amino, azo, Br, C.sub.1-8 alkyl, carbonyl,
carboxyl, Cl, cyano, C.sub.3-8 cycloalkyl, C.sub.3-8 heteroaryl,
C.sub.3-8 heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O,
phenyl, S, sulfoxide, sulfone, and/or thiophene and optionally
further substituted with acetamide, alkoxy, amino, azo, Br,
C.sub.1-3 alkyl, carbonyl, carboxyl, Cl, cyano, C.sub.3-8
cycloalkyl, C.sub.3-8 heteroaryl, C.sub.3-8 heterocyclyl, hydroxyl,
F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfone,
and/or thiophene and the alkyl, cycloalkyl, heteroaryl,
heterocyclyl, indole, or phenyl may be optionally further
substituted with one or more of the following groups: acetamide,
alkoxy, amino, azo, Br, C.sub.1-8alkyl, carbonyl, carboxyl, Cl,
cyano, C.sub.3-8 cycloalkyl, C.sub.3-8 heteroaryl,
C.sub.3-8-heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O,
phenyl, S, sulfoxide, sulfone and/or thiophene.
[0039] Optionally, the R.sup.1 group in formula (I) may be
independently substituted with one or more of the following groups:
hydroxyl; C.sub.1-10 alkyl hydroxyl (i.e., C.sub.1-C.sub.10
alkoxides); amine; C.sub.1-10 carboxylic acid; C.sub.1-10 carboxyl;
straight chain or branched C.sub.1-10 alkyl, optionally containing
unsaturation; a C.sub.2-8 cycloalkyl optionally containing
unsaturation or one oxygen or nitrogen atom; straight chain or
branched C.sub.1-10 alkyl amine; heterocyclyl; heterocyclic amine;
and/or aryl comprising phenyl, heteroaryl containing from one to
four of the following heteroatoms: N, O, and/or S, unsubstituted
phenyl ring, substituted phenyl ring, unsubstituted heterocyclyl,
and substituted heterocyclyl. Optionally, the unsubstituted phenyl
ring or substituted phenyl ring may be independently substituted
with one or more of the following groups: hydroxyl; C.sub.1-10
alkyl hydroxyl (i.e., C.sub.1-C.sub.10 alkoxides); amine;
C.sub.1-10 carboxylic acid; C.sub.1-10 carboxyl; straight chain or
branched C.sub.1-10 alkyl, optionally containing unsaturation;
straight chain or branched C.sub.1-10 alkyl amine, optionally
containing unsaturation; a C.sub.2-10 cycloalkyl optionally
containing unsaturation or one oxygen or nitrogen atom; straight
chain or branched C.sub.1-10 alkyl amine; heterocyclyl;
heterocyclic amine; and/or aryl comprising phenyl and heteroaryl
containing from one to four of the following heteroatoms: N, O,
and/or S. Optionally, the unsubstituted heterocyclyl or substituted
heterocyclyl may be independently substituted with one or more of
the following groups: hydroxyl; C.sub.1-10alkyl hydroxyl (i.e.,
C.sub.1-C.sub.10 alkoxides); amine; C.sub.1-10 carboxylic acid;
C.sub.1-10 carboxyl; straight chain or branched C.sub.1-10 alkyl
optionally containing unsaturation; straight chain or branched
C.sub.1-10 alkyl amine optionally containing unsaturation; a
C.sub.2-8 cycloalkyl optionally containing unsaturation or one
oxygen or nitrogen atom; heterocyclyl; straight chain or branched
C.sub.1-10 alkyl amine; heterocyclic amine; and/or aryl comprising
a phenyl and a heteroaryl containing from one to four of the
following heteroatoms: N, O, and S. Any of the above may be further
optionally substituted.
[0040] In some aspects, R.sup.1 in formula (I) may be optionally
substituted by one or more of the following groups: acetamide,
alkoxy, amino, azo, Br, C.sub.1-8 alkyl, carbonyl, carboxyl, Cl,
cyano, C.sub.3-8 cycloalkyl, C.sub.3-8 heteroaryl, C.sub.3-8
heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S,
sulfoxide, sulfone, and/or thiophene; and optionally further
substituted with one or more of the following groups: acetamide,
alkoxy, amino, azo, Br, C.sub.1-3 alkyl, carbonyl, carboxyl, Cl,
cyano, C.sub.3-8 cycloalkyl, C.sub.3-8 heteroaryl, C.sub.3-8
heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S,
sulfoxide, sulfone, or thiophene; wherein the alkyl, cycloalkyl,
heteroaryl, heterocyclyl, indole, or phenyl, may be optionally
further substituted with one or more of acetamide, alkoxy, amino,
azo, Br, C.sub.1-8 alkyl, carbonyl, carboxyl, Cl, cyano, C.sub.3-8
cycloalkyl, C.sub.3-8 heteroaryl, C.sub.3-8 heterocyclyl, hydroxyl,
F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide, sulfone,
and/or thiophene.
[0041] In another aspect of the disclosure, the mitofusin activator
may comprise one or more compounds represented by formula (II):
##STR00008##
or a pharmaceutically acceptable salt thereof. In Formula (II), o
may be 0, 1, 2, 3, 4, or 5; p may be 0 or 1; and q may be 0, 1, 2,
3, 4, or 5 with the proviso that the sum of o+p+q is not less than
3 or greater than 7; Z may be cycloalkyl, heterocycloalkyl, aryl,
or heteroaryl; R.sup.2 and R.sup.3 may be independently H, F,
alkyl, or C.sub.3-7 cycloalkyl; or optionally, R.sup.2 and R.sup.3
taken together may form a C.sub.3-7 cycloalkyl or heterocycloalkyl;
R.sup.4 and R.sup.5 may be independently H, F, alkyl, COR.sup.8, or
C.sub.3-7 cycloalkyl; or optionally, R.sup.4 and R.sup.5 taken
together may form a C.sub.3-7 cycloalkyl or heterocycloalkyl; Y may
be O, CR.sup.6R.sup.7, CR.sup.8.dbd.CR.sup.9, C.ident.C,
cycloalkyl, heterocycloalkyl, aryl, heteroaryl, NR.sup.8, S,
SO.sub.2, SONR.sup.9, NR.sup.9SO.sub.2, NR.sup.8CO, CONR.sup.8,
NR.sup.7CONR.sup.9; each R.sup.6 may be independently selected from
H, alkyl, and C.sub.3-7 cycloalkyl; each R.sup.7 may be
independently selected from H, alkyl, COR.sup.8 and C.sub.3-7
cycloalkyl; or optionally, R.sup.6 and R.sup.7 taken together may
form C.sub.3-7 cycloalkyl; and each R.sup.8 may be independently
selected from H, alkyl, and C.sub.3-7 cycloalkyl; each R.sup.9 may
be independently selected from H, alkyl, COR.sup.8 and C.sub.3-7
cycloalkyl; or optionally, R.sup.8 and R.sup.9 may be taken
together to form C.sub.3-7 cycloalkyl.
[0042] In some aspects, in the mitofusin activator of formula (II),
o may be 0, 1, 2, 3, 4, or 5; p may be 0 or 1; and q may be 0, 1,
2, 3, 4, or 5 with the proviso that the sum of o+p+q is not less
than 3 or greater than 7; Z may be cycloalkyl, heterocycloalkyl,
aryl, or heteroaryl; Y may be O, CR.sup.6R.sup.7, cycloalkyl, or
aryl; and R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6 and R.sup.7
may be independently selected from H or alkyl.
[0043] In some aspects, in the mitofusin activator of formula (II),
o may be 0, 1, 2, 3, 4, or 5; p may be 0 or 1; and q may be 0, 1,
2, 3, 4, or 5 with the proviso that the sum of o+p+q is not less
than 3 or greater than 5; Z may be aryl or heteroaryl; Y may be O,
CH.sub.2, or cycloalkyl; R.sup.2, R.sup.3, R.sup.4 and R.sup.5 may
each be H.
[0044] In some aspects, in the mitofusin activator of formula (II),
o may be 0, 1, 2, or 3; p may be 1; and q may be 0, 1, 2, or 3 with
the proviso that the sum of o+p+q is not less than 3 or greater
than 5; Z may be aryl or heteroaryl; Y may be cyclopropyl or
cyclobutyl; R.sup.2, R.sup.3, R.sup.4 and R.sup.5 may each be
H.
[0045] In some aspects, in the mitofusin activator of formula (II),
Z may be aryl or heteroaryl; Y may be 0 or CH.sub.2; R.sup.2,
R.sup.3, R.sup.4 and R.sup.5 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 with the proviso that
the sum of o+p+q is 5.
[0046] In some aspects, in the mitofusin activator of formula (II),
Z may be phenyl or heteroaryl, wherein the heteroaryl may contain
from one to four heteroatoms independently selected from N, O, and
S, and wherein the phenyl or heteroaryl may be substituted with
zero to four of the following independently-chosen substituents:
R.sup.8, OR.sup.8, Cl, F, CN, CF.sub.3, NR.sup.8R.sup.9,
SO.sub.2NR.sup.8R.sup.9, NR.sup.8SO.sub.2R.sup.10, SO.sub.2R.sup.9,
CONR.sup.8R.sup.10, NR.sup.8COR.sup.10, C.sub.3-7 cycloalkyl,
and/or heterocycloalkyl; each R.sup.8 may be independently selected
from H, alkyl, and C.sub.3-7 cycloalkyl; each R.sup.9 may be
independently selected from H, alkyl, COR.sup.7 and C.sub.3-7
cycloalkyl; or optionally, R.sup.8 and R.sup.9 taken together may
form a C.sub.3-7 cycloalkyl; Y may be O or CH.sub.2; R.sup.2,
R.sup.3, R.sup.4, and R.sup.5 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 with the proviso
that the sum of o+p+q is 5.
[0047] In some aspects, in the mitofusin activator of formula (II),
Z may be phenyl or heteroaryl, wherein the heteroaryl may contain
one to three of the following heteroatoms: N, O, and S, and wherein
the phenyl or heteroaryl may contain zero to three of the following
substituents independently selected from R.sup.8, OR.sup.8, Cl, F,
CN, CF.sub.3, NR.sup.8R.sup.9, SO.sub.2R.sup.9, CONR.sup.8R.sup.10,
NR.sup.8COR.sup.10, C.sub.3-7 cycloalkyl, and/or heterocycloalkyl;
Y may be O or CH.sub.2; R.sup.2, R.sup.3, R.sup.4, and R.sup.5 may
each be H; each R.sup.8 may independently be selected from H,
alkyl, and C.sub.3-7 cycloalkyl; each R.sup.9 may be independently
selected from H, alkyl, COR.sup.7 and C.sub.3-7 cycloalkyl; or
optionally, R.sup.8 and R.sup.9 taken together may form a C.sub.3-7
cycloalkyl; each R.sup.9 may be alkyl or C.sub.3-7 cycloalkyl; o
may be 0, 1, 2, 3, or 4; p may be 1; and q may be 0, 1, 2, 3, or 4
with the proviso that the sum of o+p+q is 5.
[0048] In some aspects, in the mitofusin activator of formula (II),
Z may be phenyl, 2-pyridinyl, 3-pyridinyl, 4-pyridinyl,
6-pyrimidinyl, 5-pyrimidinyl, 4-pyrimidinyl, or 2-pyrimidinyl,
wherein the phenyl, pyridinyl, and pyrimidinyl moieties may
independently contain zero to two of the following
independently-chosen substituents: R.sup.8, OR.sup.8, Cl, F, CN,
CF.sub.3, NR.sup.8R.sup.9, SO.sub.2R.sup.9, CONR.sup.8R.sup.9,
and/or NR.sup.8COR.sup.10; Y may be O or CH.sub.2; R.sup.2,
R.sup.3, R.sup.4, and R.sup.5 may each be H; each R.sup.8 may
independently be H, alkyl, and C.sub.3-7 cycloalkyl; each R.sup.9
may be independently selected from H, alkyl, COR.sup.7 and
C.sub.3-7 cycloalkyl; or optionally, R.sup.8 and R.sup.9 taken
together may form a C.sub.3-7 cycloalkyl; R.sup.10 may be alkyl or
C.sub.3-7 cycloalkyl; o may be 0, 1, 2, 3, or 4; p may be 1; and q
may be 0, 1, 2, 3, or 4 with the proviso that the sum of o+p+q is
5.
[0049] Accordingly, in some aspects, in the mitofusin activator of
formula (II), Z may be cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl; R.sup.2 and R.sup.3 may be independently selected from
H, F, alkyl, and C.sub.3-7 cycloalkyl, or R.sup.2 and R.sup.3 taken
together may form a C.sub.3-7 cycloalkyl or heterocycloalkyl;
R.sup.4 and R.sup.5 may be independently selected from H, F, alkyl,
and C.sub.3-7 cycloalkyl, or R.sup.4 and R.sup.5 taken together may
form a C.sub.3-7 cycloalkyl or heterocycloalkyl; Y is O,
CR.sup.6R.sup.7; CR.sup.8CR.sup.9, C.ident.C, cycloalkyl,
heterocycloalkyl, aryl, heteroaryl, NR.sup.8, S, SO.sub.2,
SONR.sup.9, NR.sup.9SO.sub.2, NR.sup.8CO, CONR.sup.8, or
NR.sup.8CONR.sup.9; R.sup.6 may be H, F, alkyl or cycloalkyl;
R.sup.7 may be H, F, alkyl or cycloalkyl; or R.sup.6 and R.sup.7
taken together may form a C.sub.3-7 cycloalkyl or heterocycloalkyl;
R.sup.8 may be H, F, alkyl or cycloalkyl; R.sup.9 may be H, F,
alkyl or cycloalkyl; or R.sup.8 and R.sup.9 taken together may form
a C.sub.3-7 cycloalkyl; o is 0, 1, 2, 3, 4, or 5; p is 0 or 1; and
q is 0, 1, 2, 3, 4, or 5, provided that if Y is cycloalkyl,
particularly cyclopropyl or cyclobutyl, the sum of o+p+q is not
less than 3 or greater than 5, and otherwise the sum of o+p+q is
5.
[0050] In particular aspects, in the mitofusin activator of formula
(II), Z may be aryl or heteroaryl; Y may be O, CR.sup.6R.sup.7, or
cycloalkyl; R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, and
R.sup.7 may be H or alkyl; and p is 1. In other particular aspects,
Y may be cyclopropyl or cyclobutyl, or Y may be O or CH.sub.2;
R.sup.2, R.sup.3, R.sup.4, and R.sup.5 may each be H; and p is 1.
In still other particular aspects, Z may be phenyl or heteroaryl,
wherein the heteroaryl is 2-pyridinyl, 3-pyridinyl, 4-pyridinyl,
2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, or 6-pyrimidinyl,
wherein the phenyl or heteroaryl may be optionally substituted.
Optional substitutions include 0 to 4, or 0 to 3, or 0 to 2
substituents independently selected from R.sup.8, OR.sup.8, Cl, F,
CN, CF.sub.3, NR.sub.8R.sub.9, SO.sub.2NR.sub.8R.sub.9,
NR.sub.8SO.sub.2R.sub.9, SO.sub.2R.sub.9, CONR.sub.8R.sub.9,
NR.sub.8COR.sub.10, C.sub.3-7 cycloalkyl, and heterocycloalkyl.
[0051] Mitofusin activators of the present disclosure may stimulate
mitochondrial fusion, increase mitochondrial fitness, and enhance
mitochondrial subcellular transport. More particular examples of
mitofusin activators suitable for achieving one or more of these
results may include trans-stereoisomers of 6-phenylhexanamide
derivatives or a pharmaceutically acceptable salt thereof,
particularly 6-phenyl hexanamide derivatives that are N-substituted
with a trans-4-cyclohexyl group. Compositions of the present
disclosure and methods employing comprising trans-stereoisomers of
6-phenylhexanamide derivatives may contain greater than a 1:1 molar
ratio of the trans-stereoisomer relative to the cis-stereoisomer,
such as at least about 60% or greater trans-stereoisomer on a molar
basis, or about 70% or greater, or about 80% or greater, or about
90% or greater, or about 95% or greater, or about 97% or greater,
or about 99% or greater. Compositions may even comprise
trans-stereoisomer 6-phenylhexanamide derivatives that are
racemically pure.
[0052] Particular examples of trans-stereoisomer 6-phenylhexanamide
derivatives having activity for promoting mitofusin activation
include, for example,
##STR00009## ##STR00010##
[0053] In another aspect of the present disclosure, a method of
treating a disease for which a mitofusin activator is indicated may
comprise administering to a mammal in need thereof, such as a
human, a therapeutically effective amount of a compound of formula
(II)
##STR00011##
or a pharmaceutically acceptable salt thereof. In Formula (II), o
may be 0, 1, 2, 3, 4, or 5; p may be 0 or 1; and q may be 0, 1, 2,
3, 4, or 5 with the proviso that the sum of o+p+q is not less than
3 or greater than 7; Z may be cycloalkyl, heterocycloalkyl, aryl,
or heteroaryl; R.sup.2 and R.sup.3 may be independently H, F,
alkyl, or C.sub.3-7 cycloalkyl; or optionally, R.sup.2 and R.sup.3
taken together may form a C.sub.3-7 cycloalkyl or heterocycloalkyl;
R.sup.4 and R.sup.5 may be independently H, F, alkyl, COR.sup.8, or
C.sub.3-7 cycloalkyl; or optionally, R.sup.4 and R.sup.5 taken
together may form a C.sub.3-7 cycloalkyl or heterocycloalkyl; Y may
be O, CR.sup.6R.sup.7, CR.sup.8.dbd.CR.sup.9, C.ident.C,
cycloalkyl, heterocycloalkyl, aryl, heteroaryl, NR.sup.8, S,
SO.sub.2, SONR.sup.9, NR.sup.9SO.sub.2, NR.sup.8CO, CONR.sup.8,
NR.sup.7CONR.sup.9; each R.sup.6 may be independently selected from
H, alkyl, and C.sub.3-7 cycloalkyl; each R.sup.7 may be
independently selected from H, alkyl, COR.sup.8 and C.sub.3-7
cycloalkyl; or optionally, R.sup.6 and R.sup.7 taken together may
form C.sub.3-7 cycloalkyl; and each R.sup.8 may be independently
selected from H, alkyl, and C.sub.3-7 cycloalkyl; each R.sup.9 may
be independently selected from H, alkyl, COR.sup.8 and C.sub.3-7
cycloalkyl; or optionally, R.sup.8 and R.sup.9 may be taken
together to form C.sub.3-7 cycloalkyl. Any of the mitofusin
activators specified above may be employed for treating a human or
other mammal having or suspected of having a
mitochondria-associated disease, disorder or condition.
[0054] The mitochondria-associated disease, disorder or condition
may be a pheripheral nervous system (PNS) or central nervous system
(CNS) genetic or non-genetic disorder, physical damage, and/or
chemical injury. In some aspects, in the method of treating a
disease for which a mitofusin activator is indicated, the PNS or
CNS disorder may be selected from any one or a combination of: a
chronic neurodegenerative condition wherein mitochondrial fusion,
fitness, or trafficking are impaired; a disease or disorder
associated with mitofusin-1 (MFN1) or mitofusin-2 (MFN2)
dysfunction; a disease associated with mitochondrial fragmentation,
dysfunction, or dysmotility; a degenerative neuromuscular condition
such as Charcot-Marie-Tooth disease, amyotrophic lateral sclerosis
(ALS), Huntington's disease, Alzheimer's disease, Parkinson's
disease, hereditary motor and sensory neuropathy, autism, autosomal
dominant optic atrophy (ADOA), muscular dystrophy, Lou Gehrig's
disease, cancer, mitochondrial myopathy, diabetes mellitus and
deafness (DAD), Leber's hereditary optic neuropathy (LHON), Leigh
syndrome, subacute sclerosing encephalopathy, neuropathy, ataxia,
retinitis pigmentosa, and ptosis (NARP), myoneurogenic
gastrointestinal encephalopathy (MNGIE), myoclonic epilepsy with
ragged red fibers (MERRF), mitochondrial myopathy,
encephalomyopathy, lactic acidosis, stroke-like symptoms (MELAS),
mtDNA depletion, mitochondrial neurogastrointestinal
encephalomyopathy (MNGIE), dysautonomic mitochondrial myopathy,
mitochondrial channelopathy, 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 that
involve axonal disconnection.
[0055] In some aspects of the disclosure, in the method of treating
a disease for which a mitofusin activator is indicated, the
composition may further comprise a pharmaceutically acceptable
excipient.
[0056] In some aspects of the disclosure, in the method of treating
a CNS and/or PNS genetic and/or non-genetic neurodegenerative
condition, injury, damage, and/or trauma comprising administering
to the subject a therapeutically effective amount of a mitofusin
activator according to the present disclosure.
[0057] In some aspects of the disclosure, in the method of treating
a CNS or PNS genetic or non-genetic neurodegenerative condition,
injury, damage, or trauma, the subject may be diagnosed with or may
be suspected of having one or more of the following: a chronic
neurodegenerative condition wherein mitochondrial fusion, fitness,
or trafficking are impaired; a disease or disorder associated with
MFN1 or MFN2 dysfunction; a disease associated with mitochondrial
fragmentation, dysfunction, or dysmotility; a degenerative
neuromuscular condition (such as Charcot-Marie-Tooth disease, ALS,
Huntington's disease, Alzheimer's disease, Parkinson's disease);
hereditary motor and sensory neuropathy, autism, ADOA, muscular
dystrophy, Lou Gehrig's disease, cancer, mitochondrial myopathy,
DAD, LHON, Leigh syndrome, subacute sclerosing encephalopathy,
NARP, MNGIE, MERRF, MELAS, mtDNA depletion, MNGIE, dysautonomic
mitochondrial myopathy, mitochondrial Channelopathy, PDCD/PDH,
diabetic neuropathy, chemotherapy-induced peripheral neuropathy,
crush injury, SCI, TBI, stroke, optic nerve injury, and/or related
conditions that involve axonal disconnection.
[0058] The terms "imine" or "imino," as used herein, unless
otherwise indicated, include a functional group or chemical
compound containing a carbon-nitrogen double bond. The expression
"imino compound," as used herein, unless otherwise indicated,
refers to a compound that includes an "imine" or an "imino" group
as defined herein. The "imine" or "imino" group may be optionally
substituted.
[0059] The term "hydroxy," as used herein, unless otherwise
indicated, includes --OH. The "hydroxy" may be optionally
substituted (e.g., incorporated in an alkoxide, phenoxide, or
carboxylic acid ester).
[0060] The terms "halogen" and "halo", as used herein, unless
otherwise indicated, include a chlorine, chloro, Cl; fluorine,
fluoro, F; bromine, bromo, Br; and iodine, iodo, or I.
[0061] The term "acetamide," as used herein, is an organic compound
with the formula CH.sub.3CONH.sub.2. The "acetamide" may be
optionally substituted.
[0062] The term "aryl," as used herein, unless otherwise indicated,
includes a carbocyclic aromatic group. Examples of aryl groups
include, but are not limited to, phenyl, benzyl, naphthyl, and
anthracenyl. The "aryl" may be optionally substituted.
[0063] The terms "amine" and "amino", as used herein, unless
otherwise indicated, include a functional group that contains a
nitrogen atom with a lone pair of electrons and wherein one or more
hydrogen atoms have been replaced by a substituent such as, but not
limited to, an alkyl group or an aryl group. The "amine" or "amino"
group may be optionally substituted.
[0064] The term "alkyl," as used herein, unless otherwise
indicated, includes saturated monovalent hydrocarbon radicals
having straight or branched 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,
1-hexyl, 2-hexyl, 3-hexyl, 3-methylhexyl, 2,2-dimethylhexyl,
2,4-dimethylhexyl, 2,5-dimethylhexyl, 3,5-dimethylhexyl,
2,4-dimethylpentyl, 2-methylheptyl, and 3-methylheptyl. Unsaturated
alkyl groups may be referred to as alkenyl (at least one
carbon-carbon double bond) or alkynyl (at least one carbon-carbon
triple bond) groups, which may include, but are 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, acetylenyl, propynyl, -1-butynyl,
2-butynyl, 1-pentynyl, 2-pentynyl, or 3-methyl-1 butynyl. E and Z
isomers may be present in any alkenyl group. The "alkyl,"
"alkenyl," or "alkynyl" may be optionally substituted.
[0065] 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). The "carboxyl" may be optionally substituted.
[0066] The term "acyl," as used herein, unless otherwise indicated,
includes a functional group derived from an aliphatic carboxylic
acid by removal of the hydroxyl (--OH) group. The "acyl" may be
optionally substituted.
[0067] The term "alkoxy," as used herein, unless otherwise
indicated, includes O-alkyl groups wherein alkyl is as defined
above and O represents oxygen. Representative alkoxy groups
include, but are not limited to, --O-methyl, --O-ethyl,
--O-n-propyl, --O-n-butyl, --O-n-pentyl, --O-n-hexyl, --O-n-heptyl,
--O-n-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-2,4dimethylpentyl, --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-cyclopropyl, --O-cyclobutyl, --O-- cyclopentyl, --O-cyclohexyl,
--O-cycloheptyl, --O-cyclooctyl, --O-cyclononyl, --O-- cyclodecyl,
--O--CH.sub.2-cyclopropyl, --O--CH.sub.2-cyclobutyl,
--O--CH.sub.2-cyclopentyl, --O--CH.sub.2-cyclohexyl,
--O--CH.sub.2-cycloheptyl, --O--CH.sub.2-cyclooctyl,
--O--CH.sub.2-cyclononyl, --O--CH.sub.2-cyclodecyl,
--O--(CH.sub.2).sub.n-cyclopropyl,
--O--(CH.sub.2).sub.n-cyclobutyl,
--O--(CH.sub.2).sub.n-cyclopentyl,
--O--(CH.sub.2).sub.n-cyclohexyl,
--O--(CH.sub.2).sub.n-cycloheptyl,
--O--(CH.sub.2).sub.n-cyclooctyl, --O--(CH.sub.2).sub.n-cyclononyl,
and/or --O--(CH.sub.2).sub.n-cyclodecyl. The alkoxy may be
saturated, partially saturated, or unsaturated. The "alkoxy" may be
optionally substituted. In any example above, n may be from one to
about twenty.
[0068] The term "cycloalkyl," as used herein, unless otherwise
indicated, includes a non-aromatic, saturated, partially saturated,
or unsaturated, monocyclic or fused, spiro or unfused bicyclic or
tricyclic hydrocarbon referred to herein containing a total of from
3 to 10 carbon atoms. Examples of cycloalkyls include, but are not
limited to, C.sub.3-10 cycloalkyl groups including cyclopropyl,
cyclobutyl, cyclopentyl, cyclopentadienyl, cyclohexyl,
cyclohexenyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl,
cycloheptyl, 1,3-cycloheptadienyl, 1,3,5-cycloheptatrienyl,
cyclooctyl, and cyclooctadienyl. The term "cycloalkyl" also
includes lower alkyl-cycloalkyl, wherein lower alkyl and cycloalkyl
are as defined herein. Examples of -lower alkyl-cycloalkyl groups
include, but are not limited to, --CH.sub.2-cyclopropyl,
--CH.sub.2-cyclobutyl, --CH.sub.2-cyclopentyl,
--CH.sub.2-cyclopentadienyl, --CH.sub.2-cyclohexyl,
--CH.sub.2-cycloheptyl, and/or --CH.sub.2-cyclooctyl. The
"cycloalkyl" may be optionally substituted.
[0069] The term "heterocyclic" as used herein, unless otherwise
indicated, includes an aromatic group or non-aromatic cycloalkyl
group in which one to four of the ring carbon atoms are
independently replaced with one or more of O, S, and N. Aromatic
heterocyclic groups are referred to as "heteroaryl" groups.
Non-aromatic heterocyclic groups are referred to as "heterocyclyl"
groups. Representative examples of heterocyclic groups 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)-dioxolane, 4,5-dihydro-1H-imidazolyl, and/or tetrazolyl.
Heterocyclic groups may be substituted or unsubstituted.
Heterocyclic groups may also be bonded at any ring atom (i.e., at
any carbon atom or heteroatom of the heterocyclic ring). The
heterocyclic group may be saturated, partially saturated, or
unsaturated.
[0070] The term "indole," as used herein, is an aromatic
heterocyclic organic compound with formula C.sub.8H.sub.7N. It has
a bicyclic structure containing a six-membered benzene ring fused
to a five-membered nitrogen-containing pyrrole ring. The "indole"
may be optionally substituted.
[0071] The term "cyano," as used herein, unless otherwise
indicated, includes a --CN group.
[0072] The term "alcohol," as used herein, unless otherwise
indicated, includes a compound in which a hydroxy functional group
(--OH) is bound to a carbon atom. In particular, this carbon atom
may be saturated, having single bonds to three other atoms. The
"alcohol" may be optionally substituted. The "alcohol" may be a
primary, secondary, or tertiary alcohol.
[0073] The term "solvate" is intended to mean a solvated form of a
specified compound that retains the effectiveness of such compound.
Examples of solvates include compounds of the invention in
combination with, but not limited to, one or more of: water,
isopropanol, ethanol, methanol, dimethylsulfoxide (DMSO), ethyl
acetate, acetic acid, or ethanolamine.
[0074] The term "stereoisomer" encompasses both optical isomers,
such as enantiomers or diastereomers, the latter existing due to
more than one center of chirality in the molecule, as well as
geometrical isomers (cis/trans isomers or diastereomers).
[0075] A composition comprising the trans-stereoisomer
6-phenylhexanamide derivative mitofusin activator or a
pharmaceutically acceptable salt thereof of the disclosure may
comprise the trans-stereoisomer in greater amount than the
cis-stereoisomer. A method of using the composition comprising the
trans-stereoisomer 6-phenylhexanamide derivative mitofusin
activator or a pharmaceutically acceptable salt thereof of the
disclosure may be such that the composition comprises the
trans-stereoisomer in greater amount than the cis-stereoisomer.
[0076] The term "mmol," as used herein, is intended to mean
millimole. The term "equiv" and "eq.," as used herein, are intended
to mean equivalent. The term "mL," as used herein, is intended to
mean milliliter. The term "g," as used herein, is intended to mean
gram. The term "kg," as used herein, is intended to mean kilogram.
The term ".mu.g," as used herein, is intended to mean micrograms.
The term "h," as used herein, is intended to mean hour. The term
"min," as used herein, is intended to mean minute. The term "M," as
used herein, is intended to mean molar. The term ".mu.L," as used
herein, is intended to mean microliter. The term ".mu.M," as used
herein, is intended to mean micromolar. The term "nM," as used
herein, is intended to mean nanomolar. The term "N," as used
herein, is intended to mean normal. The term "amu," as used herein,
is intended to mean atomic mass unit. The term ".degree. C.," as
used herein, is intended to mean degree Celsius. The term "wt/wt,"
as used herein, is intended to mean weight/weight. The term "v/v,"
as used herein, is intended to mean volume/volume. The term "MS,"
as used herein, is intended to mean mass spectrometry. The term
"HPLC," as used herein, is intended to mean high performance liquid
chromatography. The term "RT," as used herein, is intended to mean
room temperature or retention time, depending on context. The term
"e.g.," as used herein, is intended to mean for example. The term
"N/A," as used herein, is intended to mean not tested or not
applicable.
[0077] As used herein, the expression "pharmaceutically acceptable
salt" refers to pharmaceutically acceptable organic or inorganic
salts of a compound of the 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, glucaronate, saccharate, formate,
benzoate, glutamate, methanesulfonate, ethanesulfonate,
benzenesulfonate, p-toluenesulfonate, and/or pamoate (i.e.,
1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts. A
pharmaceutically acceptable salt may involve the inclusion of
another molecule such as an acetate ion, a succinate ion, or other
counterion. The counterion may 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. Instances where multiple charged atoms are
part of the pharmaceutically acceptable salt may have multiple
counterions. Hence, a pharmaceutically acceptable salt may have one
or more charged atoms and/or one or more counterion. As used
herein, the expression "pharmaceutically acceptable solvate" refers
to an association of one or more solvent molecules and a compound
of the 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 expression
"pharmaceutically acceptable hydrate" refers to a compound of the
invention, or a salt thereof, that further includes a
stoichiometric or non-stoichiometric amount of water bound by
non-covalent intermolecular forces.
[0078] Each of the states, diseases, disorders, and conditions,
described herein, as well as others, can benefit from compositions
and methods described herein. Generally, treating a state, disease,
disorder, or condition includes preventing or delaying the
appearance of clinical symptoms in a mammal that may be afflicted
with or predisposed to the state, disease, disorder, or condition
but does not yet experience or display clinical or subclinical
symptoms thereof. Treating can also include inhibiting the state,
disease, disorder, or condition, e.g., arresting or reducing the
development of the disease or at least one clinical or subclinical
symptom thereof. Furthermore, treating can include relieving the
disease, e.g., causing regression of the state, disease, disorder,
or condition or at least one of its clinical or subclinical
symptoms. A benefit to a subject to be treated can be either
statistically significant or at least perceptible to the subject or
to a physician.
Mitofusin 1 and Mitofusin 2
[0079] Mitofusins (MFN) 1 and 2, named for their central roles in
mitochondrial fusion, are attractive drug targets because their
regulatory functioning in mitochondrial dynamics and
quality/quantity control is perturbed in several neurodegenerative
disorders. In particular, genetic mutations that abrogate or impair
MFN2 functioning and therefore suppress mitochondrial fusion cause
the rare autosomal dominant neurodegenerative condition
Charcot-Marie-Tooth disease type 2A (CMT2A), for which there is
currently no disease-altering therapy. Moreover, accumulating
experimental evidence supports an important role for MFN1 and MFN2
in heart disease. A pharmacological means of enhancing mitofusin
function has the potential to correct the underlying cause of CMT2A
and other cardiac or neurodegenerative diseases caused by
mitochondrial dys-dynamism. MFN1 and MFN2 form homo (MFN1-MFN1 or
MFN2-MFN2) or hetero (MFN1-MFN2) trans-dimers between mitochondria.
This process is referred to as mitochondrial tethering, and is the
requisite first step in mitochondrial fusion essential for
metabolic cellular health (Koshiba, T., et al.; Science 305:858-62,
2004). Mitochondrial tethering and fusion depend upon a shift in
MFN protein conformation, from a more closed resting state to a
more open active state. MFN conformation is regulated by
intra-molecular peptide-peptide interactions (PPI) between 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 tight alpha
helix within the interacting peptide, directing critical Va372, Met
376, and His380 amino acid side chains to their interacting
partners Leu727, Leu723, and Lys720 respectively (Rocha, A. G., et
al.; Science 360:336-41, 2018). A strong PPI promotes a folded
protein conformation that decreases the probability of trans
MFN-MFN dimer formation between mitochondria (e.g., is unfavorable
for mitochondrial tethering/fusion). When serine 378 is not
phosphorylated the interacting peptide alpha helix partially
unwinds, weakening the PPI and increasing the probability that the
protein will unfold to permit MFN trans-dimerization and subsequent
mitochondrial fusion (Rocha, A. G., et al.; Science 2018).
[0080] Franco, et al. (Franco, A., et al.; Nature 2016) described
an 18 amino acid peptide, modified slightly from hMFN2 amino acids
367-384, that competitively inhibited intra-molecular PPI in MFN1
and MFN2, thus promoting the protein conformation favoring
mitochondrial fusion. By understanding the critical interacting
amino acids within this activator peptide, a pharmacophore model
was developed leading to identification of a prototype small
molecule mitofusin activator, Chimera B-A/I, which mimicked
mitofusin activator peptide effects by promoting mitochondrial
fusion after topical application to mitofusin deficient cultured
cells and by increasing mitochondrial motility in ex vivo CMT2A
nerves (Rocha, A. G., et al.; Science 2018). Clinical application
of small molecule mitofusin activators having drug-like properties
promises the first disease-altering therapy for CMT2A and may open
the door to a novel therapeutic approach of enhancing mitochondrial
fusion in multiple diseases with impaired mitochondrial
dynamics.
Mitochondria-Associated Diseases, Disorders, or Conditions
[0081] The present disclosure provides for compositions and methods
of treatment for treating mitochondria-related diseases, disorders,
or conditions, including diseases or disorders associated with MFN1
and/or MFN2 and mitochondrial dysfunction. A
mitochondria-associated disease, disorder, or condition may be a
disease primarily caused by or secondarily associated with
mitochondrial dysfunction, fragmentation, or loss-of-fusion, or
associated with dysfunction in MFN1 or MFN2 catalytic activity or
conformational unfolding. Mitochondrial dysfunction may be caused
by genetic mutations of mitofusins or other (nuclear or
mitochondrial encoded) genes, or may be caused by physical,
chemical, or environmental injury to the CNS or PNS.
[0082] Mitochondria transit within cells and undergo fusion to
exchange genomes and promote mutual repair. Mitochondrial fusion
and subcellular trafficking are mediated in part by MFN1 and MFN2.
Genetic mutations in MFN2 that suppress mitochondrial fusion and
motility cause Charcot-Marie-Tooth Disease, type 2A (CMT2A), the
most common heritable axonal neuropathy. Mitochondrial
fragmentation, dysfunction, and dysmotility are also central
features of other genetic neurodegenerative syndromes, such as
amyotrophic lateral sclerosis, Huntington's disease, Parkinson's
disease, and Alzheimer's disease. Because no therapeutics exist
that directly enhance mitochondrial fusion or trafficking, these
diseases are unrelenting and considered irreversible.
[0083] Examples of mitochondria-associated 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 neuropathy,
autism, ADOA, muscular dystrophy, Lou Gehrig's disease, cancer,
mitochondrial myopathy, DAD, LHON, Leigh syndrome, subacute
sclerosing encephalopathy, NARP, MNGIE, MERRF, MELAS, mtDNA
depletion, MNGIE, dysautonomic mitochondrial myopathy,
mitochondrial channelopathy, and/or PDCD/PDH.
[0084] Symptoms that may be treated with the methods as described
herein include, but are not limited to, poor growth, loss of muscle
coordination, muscle paralysis and atrophy, visual problems,
hearing problems, learning disabilities, heart disease, liver
disease, kidney disease, gastrointestinal disorders, respiratory
disorders, neurological problems, autonomic dysfunction, and
dementia.
Neurodegenerative Disease
[0085] As described herein, trans-stereoisomer 6-phenylhexanamide
derivative mitofusin activators may rapidly reverse mitochondrial
dysmotility in sciatic nerve axons of a mouse model of
Charcot-Marie-Tooth disease, type 2A. Because impaired
mitochondrial fusion, fitness, and/or trafficking also contribute
to neuronal degeneration in various neurodegenerative diseases
(e.g., in Charcot-Marie-Tooth disease (CMT2A), Huntington's
disease, Parkinson's disease, and Alzheimer's disease, and
especially in ALS), the present disclosure provides for
compositions (e.g., compositions containing mitofusin activators)
and methods to treat such neurodegenerative diseases, disorders,
and/or conditions.
[0086] Examples of neurodegenerative diseases, disorders and
conditions include a disease of impaired neuronal mitochondrial
dynamism or trafficking, such as, but not limited to, a hereditary
motor and sensory neuropathy (HMSN) (e.g., CMT1 (a dominantly
inherited, hypertrophic, predominantly demyelinating form), CMT2 (a
dominantly inherited predominantly axonal form), Dejerine-Sottas
(severe form with onset in infancy), CMTX (inherited in an X-linked
manner), and CMT4 (includes the various demyelinating autosomal
recessive forms of Charcot-Marie-Tooth disease); hereditary sensory
and autonomic neuropathy type IE, hereditary sensory and autonomic
neuropathy type II, hereditary sensory and autonomic neuropathy
type V, HMSN types 1A and 1B (e.g., dominantly inherited
hypertrophic demyelinating neuropathies), HMSN type 2 (e.g.,
dominantly inherited neuronal neuropathies), HMSN type 3 (e.g.,
hypertrophic neuropathy of infancy [Dejerine-Sottas]), HMSN type 4
(e.g., hypertrophic neuropathy [Refsum] associated with phytanic
acid excess), HMSN type 5 (associated with spastic paraplegia),
and/or HMSN type 6 (e.g., with optic atrophy)).
[0087] Other examples of neurodegenerative diseases, disorders, and
conditions include, but are not limited to, Alzheimer's disease,
ALS, Alexander disease, Alpers' disease, Alpers-Huttenlocher
syndrome, alpha-methylacyl-CoA racemase deficiency, Andermann
syndrome, Arts syndrome, ataxia neuropathy spectrum, ataxia (e.g.,
with oculomotor apraxia, autosomal dominant cerebellar ataxia,
deafness, and narcolepsy), autosomal recessive spastic ataxia of
Charlevoix-Saguenay, Batten disease, beta-propeller
protein-associated neurodegeneration, cerebro-oculo-facio-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 with anhidrosis, dementia, familial
encephalopathy with neuroserpin inclusion bodies, familial British
dementia, familial Danish dementia, fatty acid
hydroxylase-associated neurodegeneration, Friedreich's Ataxia,
Gerstmann-Straussler-Scheinker Disease, GM2-gangliosidosis (e.g.,
AB variant), HMSN type 7 (e.g., with retinitis pigmentosa),
Huntington's disease, infantile neuroaxonal dystrophy,
infantile-onset ascending hereditary spastic paralysis,
infantile-onset spinocerebellar ataxia, juvenile primary lateral
sclerosis, Kennedy's disease, Kuru, Leigh's Disease,
Marinesco-Sjogren syndrome, mild cognitive impairment (MCI),
mitochondrial membrane protein-associated neurodegeneration, motor
neuron disease, monomelic amyotrophy, motor neuron diseases (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 of aging, neurodegeneration with brain iron
accumulation, neuromyelitis optica, pantothenate kinase-associated
neurodegeneration, opsoclonus myoclonus, prion disease, progressive
multifocal leukoencephalopathy, Parkinson's disease, Parkinson's
disease-related disorders, polycystic lipomembranous osteodysplasia
with sclerosing leukoencephalopathy, prion disease, progressive
external ophthalmoplegia, riboflavin transporter deficiency
neuronopathy, Sandhoff disease, spinal muscular atrophy (SMA),
spinocerebellar ataxia (SCA), striatonigral degeneration,
transmissible spongiform encephalopathies (prion diseases), and/or
Wallerian-like degeneration.
Charcot-Marie-Tooth (CMT) Disease Type 2A
[0088] Charcot-Marie-Tooth type 2A (CMT2A) disease is an example of
a non-curable neurodegenerative disease/axonal neuropathy,
disorder, or condition caused by mutations of MFN2 and for which
there are currently no disease-modifying treatments. As described
herein, it was discovered that severely impaired mitochondrial
transport from neuron cell body in the spinal cord to distal
neuronal synapse in the lower leg or hand (in addition to smaller
mitochondria size as is widely recognized) is a central factor in
CMT2A disease onset and progression. CMT2A is a progressive
neuromuscular disease that typically causes muscle weakness and
wasting in the distal legs/feet in children of ages 1-8 years, then
upper limbs, ultimately producing severe muscle wasting, skeletal
deformities, and permanent disability. The present disclosure
provides for the correction of impaired neuronal mitochondria
transport as a therapeutic target in this disease. Data showed that
administration of a trans-6-phenylhexanamide mitofusin activators
promoted the mitochondria to move along neuronal axons in mouse
models where mitochondria were not previously moving, and reversed
disease-associated defects in neuromuscular function, which is
applicable in any neuropathy (e.g., Huntington's disease, ALS,
ALS-like sclerosis, and/or Alzheimer's disease).
Neurological and Neurodegenerative Diseases
[0089] As described herein, trans-4-hydroxycyclohexyl
6-phenylhexanamide derivative mitofusin activators may rapidly
reverse mitochondrial dysmotility in sciatic nerve axons of a mouse
model of Charcot-Marie-Tooth disease type 2A. It is currently
believed that impaired mitochondrial trafficking also contribute to
neuronal degeneration in various neurological diseases (e.g., in
Huntington's disease, Parkinson's disease, and Alzheimer's disease,
and especially in ALS). As such, the present disclosure provides
for methods and compositions to treat neurological diseases,
disorders, or conditions. For example, a neurological disease,
disorder, or condition may be, but is not limited to, abulia;
agraphia; alcoholism; alexia; alien hand syndrome;
Allan-Herndon-Dudley syndrome; alternating hemiplegia of childhood;
Alzheimer's disease; amaurosis fugax; amnesia; ALS; aneurysm;
angelman syndrome; anosognosia; aphasia; apraxia; arachnoiditis;
Arnold-Chiari malformation; asomatognosia; Asperger syndrome;
ataxia; attention deficit hyperactivity disorder; atr-16 syndrome;
auditory processing disorder; autism spectrum; Behcets disease;
bipolar disorder; Bell's palsy; brachial plexus injury; brain
damage; brain injury; brain tumor; Brody myopathy; Canavan disease;
capgras delusion; carpal tunnel syndrome; causalgia; central pain
syndrome; central pontine myelinolysis; centronuclear myopathy;
cephalic disorder; cerebral aneurysm; cerebral arteriosclerosis;
cerebral atrophy; cerebral autosomal dominant arteriopathy with
subcortical infarcts and leukoencephalopathy (CADASIL); cerebral
dysgenesis-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; compression neuropathy; congenital facial diplegia;
corticobasal degeneration; cranial arteritis; craniosynostosis;
Creutzfeldt-Jakob disease; cumulative trauma disorders; Cushing's
syndrome; cyclothymic disorder; cyclic vomiting syndrome (CVS);
cytomegalic inclusion body disease (CIBD); cytomegalovirus
infection; Dandy-Walker syndrome; dawson disease; de Morsier's
syndrome; Dejerine-Klumpke palsy; Dejerine-Sottas disease; delayed
sleep phase syndrome; dementia; dermatomyositis; developmental
coordination disorder; diabetic neuropathy; diffuse sclerosis;
diplopia; disorders of consciousness; down syndrome; Dravet
syndrome; duchenne muscular dystrophy; dysarthria; dysautonomia;
dyscalculia; dysgraphia; dyskinesia; dyslexia; dystonia; empty
sella syndrome; encephalitis; encephalocele; encephalotrigeminal
angiomatosis; encopresis; enuresis; epilepsy; epilepsy-intellectual
disability in females; erb's palsy; erythromelalgia; essential
tremor; exploding head syndrome; Fabry's disease; Fahr's syndrome;
fainting; familial spastic paralysis; febrile seizures; Fisher
syndrome; Friedreich's ataxia; fibromyalgia; Foville's syndrome;
fetal alcohol syndrome; fragile x syndrome; fragile x-associated
tremor/ataxia syndrome (FXTAS); Gaucher's disease; generalized
epilepsy with febrile seizures plus; Gerstmann's syndrome; giant
cell arteritis; giant cell inclusion disease; globoid cell
leukodystrophy; gray matter heterotopia; Guillain-Barre syndrome;
generalized anxiety disorder; HTLV-1 associated myelopathy;
Hallervorden-Spatz syndrome; head injury; headache; hemifacial
spasm; hereditary spastic paraplegia; heredopathia atactica
polyneuritiformis; herpes zoster oticus; herpes zoster; Hirayama
syndrome; Hirschsprung's disease; Holmes-Adie syndrome;
holoprosencephaly; Huntington's disease; hydranencephaly;
hydrocephalus; hypercortisolism; hypoxia; immune-mediated
encephalomyelitis; inclusion body myositis; incontinentia pigmenti;
infantile refsum disease; infantile spasms; inflammatory myopathy;
intracranial cyst; intracranial hypertension; isodicentric 15;
Joubert syndrome; Karak syndrome; Kearns-Sayre syndrome; Kinsbourne
syndrome; Kleine-Levin syndrome; Klippel Feil syndrome; Krabbe
disease; Kufor-Rakeb syndrome; Lafora disease; Lambert-Eaton
myasthenic syndrome; Landau-Kleffner syndrome; lateral medullary
(Wallenberg) syndrome; learning disabilities; Leigh's disease;
Lennox-Gastaut syndrome; Lesch-Nyhan syndrome; leukodystrophy;
leukoencephalopathy with vanishing white matter; lewy body
dementia; lissencephaly; locked-in syndrome; Lou Gehrig's disease
(amyotrophic lateral sclerosis (ALS)); lumbar disc disease; lumbar
spinal stenosis; lyme disease--neurological sequelae;
Machado-Joseph disease (spinocerebellar ataxia type 3);
macrencephaly; macropsia; mal de debarquement; megalencephalic
leukoencephalopathy with subcortical cysts; megalencephaly;
Melkersson-Rosenthal syndrome; menieres disease; meningitis; Menkes
disease; metachromatic leukodystrophy; microcephaly; micropsia;
migraine; Miller Fisher syndrome; mini-stroke (transient ischemic
attack); misophonia; mitochondrial myopathy; mobius syndrome;
monomelic amyotrophy; Morvan syndrome; motor neurone disease--see
ALS; motor skills disorder; moyamoya disease;
mucopolysaccharidoses; multi-infarct dementia; multifocal motor
neuropathy; multiple sclerosis; multiple system atrophy; muscular
dystrophy; myalgic encephalomyelitis; myasthenia gravis;
myelinoclastic diffuse sclerosis; myoclonic encephalopathy of
infants; myoclonus; myopathy; myotubular myopathy; myotonia
congenita; narcolepsy; neuro-Behcet's disease; neurofibromatosis;
neuroleptic malignant syndrome; neurological manifestations of
aids; neurological sequelae of lupus; neuromyotonia; neuronal
ceroid lipofuscinosis; neuronal migration disorders; neuropathy;
neurosis; Niemann-Pick disease; non-24-hour sleep-wake disorder;
nonverbal learning disorder; O'Sullivan-McLeod syndrome; occipital
neuralgia; occult spinal dysraphism sequence; Ohtahara syndrome;
olivopontocerebellar atrophy; opsoclonus myoclonus syndrome; optic
neuritis; orthostatic hypotension; otosclerosis; overuse syndrome;
palinopsia; paresthesia; Parkinson's disease; paramyotonia
congenita; paraneoplastic diseases; paroxysmal attacks;
Parry-Romberg syndrome; pediatric autoimmune neuropsychiatric
disorders associated with streptococcoal infections (PANDAS);
Pelizaeus-Merzbacher disease; periodic paralyses; peripheral
neuropathy; pervasive developmental disorders; phantom limb/phantom
pain; photic sneeze reflex; phytanic acid storage disease; Pick's
disease; pinched nerve; pituitary tumors; pmg; polyneuropathy;
polio; polymicrogyria; polymyositis; porencephaly; post-polio
syndrome; postherpetic neuralgia (phn); postural hypotension;
Prader-Willi syndrome; primary lateral sclerosis; prion diseases;
progressive hemifacial atrophy; progressive multifocal
leukoencephalopathy; progressive supranuclear palsy; prosopagnosia;
pseudotumor cerebri; quadrantanopia; quadriplegia; rabies;
radiculopathy; Ramsay Hunt syndrome type 1; Ramsay Hunt syndrome
type 2; Ramsay Hunt syndrome type 3--see Ramsay-Hunt syndrome;
Rasmussen encephalitis; reflex neurovascular dystrophy; refsum
disease; REM sleep behavior disorder; repetitive stress injury;
restless legs syndrome; retrovirus-associated myelopathy; Rett
syndrome; Reye's syndrome; rhythmic movement disorder; Romberg
syndrome; Saint Vitus' dance; Sandhoff disease; Schilder's disease
(two distinct conditions); schizencephaly; sensory processing
disorder; septo-optic dysplasia; shaken baby syndrome; shingles;
Shy-Drager syndrome; Sjogren's syndrome; sleep apnea; sleeping
sickness; snatiation; Sotos syndrome; spasticity; spina bifida;
spinal cord injury; spinal cord tumors; spinal muscular atrophy;
spinal and bulbar muscular atrophy; spinocerebellar ataxia;
split-brain; Steele-Richardson-Olszewski syndrome; stiff-person
syndrome; stroke; Sturge-Weber syndrome; stuttering; subacute
sclerosing panencephalitis; subcortical arteriosclerotic
encephalopathy; superficial siderosis; Sydenham's chorea; syncope;
synesthesia; syringomyelia; tarsal tunnel syndrome; tardive
dyskinesia; tardive dysphrenia; Tarlov cyst; Tay-Sachs disease;
temporal arteritis; temporal lobe epilepsy; tetanus; tethered
spinal cord syndrome; Thomsen disease; thoracic outlet syndrome;
tic douloureux; Todd's Paralysis; tourette syndrome; toxic
encephalopathy; transient ischemic attack; transmissible spongiform
encephalopathies; 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); viliuisk
encephalomyelitis (VE); Wallenberg's syndrome; west syndrome;
whiplash; Williams syndrome; Wilson's disease; y-linked hearing
impairment; and/or Zellweger syndrome.
Chemotherapy-Induced Peripheral Neuropathy (CIPN)
[0090] Cancer chemotherapy-induced sensory and motor neuropathies
may be prevented or treated with the compostions and methods
described herein. Although cancer continues to be a leading cause
of mortality world-wide, early detection and improved cancer
chemotherapeutics preferentially attacking rapidly dividing cells
are favorably impacting this disease. Consequently, the number of
cancer survivors is increasing and collateral detrimental effects
of successful cancer therapy are a growing problem for cancer
survivors. Chemotherapy-induced peripheral neuropathy is one of the
most common complications of cancer chemotherapy, affecting 20% of
all patients and almost 100% of patients receiving high doses of
chemotherapeutic agents. Dose-dependent neurotoxicity of motor and
sensory neurons can lead to chronic pain, hypersensitivity to hot,
cold, and mechanical stimuli, and/or impaired neuromuscular
control. The most common chemotherapeutic agents linked to CIPN are
platinum, vinca alkaloids, taxanes, epothilones, and the targeted
proteasome inhibitor, bortezomib.
[0091] CIPN most commonly affects peripheral sensory neurons whose
cell bodies are located in dorsal root ganglia lacking the
blood-brain barrier that protects other components of the central
and peripheral nervous system. Unprotected dorsal root ganglion
neurons are more sensitive to neuronal hyperexcitability and innate
immune system activation evoked by circulating cytotoxic
chemotherapeutic agents. CIPN affects quality of life, and is
potentially disabling, because it provokes chronic neuropathic pain
that, like other causes of neuralgia (e.g., post herpetic
neuralgia, diabetic mononeuropathy), is refractory to analgesic
therapy. Motor nerve involvement commonly manifests as loss of fine
motor function with deterioration in hand writing, difficulty in
buttoning clothes or sewing, and sometimes upper and lower
extremity weakness or loss of endurance. CIPN typically manifests
within weeks of chemotherapy and in many cases improves after
chemotherapy treatment ends, although residual pain, sensory, or
motor defects are observed in one third to one half of affected
patients. Unfortunately, CIPN-limited chemotherapy dosing can lead
to delays, reduction, or interruption of cancer treatment, thus
shortening survival.
[0092] Mitochondrial dysfunction and oxidative stress are
implicated in CIPN because of observed ultrastructural
morphological abnormalities, impaired mitochondria DNA
transcription and replication, induction of mitochondrial apoptosis
pathways, and reduction of experimental CIPN signs by anticipatory
mitochondrial protection. As described herein,
trans-4-hydroxycyclohexyl 6-phenylhexanamide derivative mitofusin
activators may enhance overall mitochondrial function in damaged
neurons, increase mitochondrial transport to areas of neuronal
damage, and accelerate in vitro neuron repair/regeneration after
chemotherapy-induced damage. For this reason, it is believed that
trans-4-hydroxycyclohexyl 6-phenylhexanamide derivative mitofusin
activators may reduce neuronal injury conferred by chemotherapeutic
agents in CIPN and accelerate regeneration/repair of nerves damaged
by chemotherapeutic anticancer agents. Further testing of the CIPN
damage prevention/repair and regeneration hypothesis will be
further developed with the trans-4-hydroxycyclohexyl
6-phenylhexanamide derivative mitofusin activators by evaluating
their in vivo effectiveness. As such, the present disclosure
provides for compositions and methods to treat cancer chemotherapy
induced nerve injury and neuropathy.
CNS or PNS Injury or Trauma
[0093] Injury in the CNS or PNS (e.g., trauma to the CNS or PNS,
crush injury, SCI, TBI, stroke, optic nerve injury, or related
conditions that involve axonal disconnection) may be treated with
the compositions and methods as described herein. The CNS includes
the brain and the spinal cord and the PNS is composed of cranial,
spinal, and autonomic nerves that connect to the CNS.
[0094] Damage to the nervous system caused by mechanical, thermal,
chemical, or ischemic factors may impair various nervous system
functions such as memory, cognition, language, and voluntary
movement. Most often, this is through accidental crush or
transection of nerve tracts, or as an unintended consequence of
medical interventions, that interrupt normal communications between
nerve cell bodies and their targets. Other types of injuries may
include disruption of the interrelations between neurons and their
supporting cells or the destruction of the blood-brain barrier.
[0095] As described herein, trans-4-hydroxycyclohexyl
6-phenylhexanamide derivative mitofusin activators may rapidly
reverse mitochondrial dysmotility in neurons from mice or patients
with various genetic or chemotherapeutic neurodegenerative
diseases, in axons injured by chemotherapeutic agents, and in axons
severed by physical injury. For this reason, it is believed that
enhancing mitochondrial trafficking with trans-4-hydroxycyclohexyl
6-phenylhexanamide derivative mitofusin activators may enhance
regeneration/repair of physically damaged nerves, as in vehicular
and sports injuries, penetration trauma from military or criminal
actions, and iatrogenic injury during invasive medical procedures.
Further testing of the injury-regeneration hypothesis will be
further developed with the small molecule mitofusin activators for
evaluation of their in vivo effectiveness. As such, the present
disclosure provides for compositions and methods to treat physical
nerve injury.
[0096] As disclosed herein, mitochondrial motility is implicated in
neuropathy. It is believed that mitochondrial motility is also
implicated in traumatic crush or severance nerve injuries. After
nerve laceration or crush injury, nerves will either regenerate and
restore neuromuscular function or fail to regenerate such that
neuromuscular function in permanently impaired.
Trans-4-hydroxycyclohexyl 6-phenylhexanamide derivative mitofusin
activators, as described herein, may increase mitochondrial
trafficking, enabling the nerve to regenerate after traumatic
injuries.
Formulation
[0097] The agents and compositions described herein may be
formulated by any conventional manner using one or more
pharmaceutically acceptable carriers or excipients as described
previously (e.g., Remington's Pharmaceutical Sciences (A. R.
Gennaro, Ed.), 21st edition, ISBN: 0781746736 (2005), which is
incorporated herein by reference with respect to its disclosure of
pharmaceutically acceptable carriers). Such formulations will
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 to provide the form for proper
administration to a subject.
[0098] The term "formulation" refers to a preparation of a drug in
a form suitable for administration to a subject such as a human or
animal pet or livestock. Thus, a "formulation" may include
pharmaceutically acceptable excipients, including diluents or
carriers.
[0099] The term "pharmaceutically acceptable," as used herein,
describes substances or components that do not cause unacceptable
losses of pharmacological activity or unacceptable adverse side
effects. One of skill in the art will be familiar with suitable
pharmaceutically acceptable substances. Examples of
pharmaceutically acceptable ingredients include those having
monographs in United States Pharmacopeia (USP 29) and National
Formulary (NF 24), United States Pharmacopeial Convention, Inc,
Rockville, Md., 2005 ("USP/NF"), or a more recent edition, and the
components listed in the continuously updated Inactive Ingredient
Search online database of the FDA. Other useful components that are
not described in the USP/NF may also be used.
[0100] The term "pharmaceutically acceptable excipient," as used
herein, includes solvents, dispersion media, coatings,
antibacterial agents, antifungal agents, isotonic, and absorption
delaying agents. The use of such media and agents for
pharmaceutical active substances is well known in the art (see
generally Remington's Pharmaceutical Sciences (A. R. Gennaro, Ed.),
21st edition, ISBN: 0781746736 (2005)). Except insofar as any
conventional media or agent is incompatible with an active
ingredient, its use in the therapeutic compositions is
contemplated. Supplementary active ingredients may also be
incorporated into the compositions.
[0101] A "stable" formulation or composition refers to a
composition having sufficient stability to allow storage at a
convenient temperature, such as between about 0.degree. C. and
about 60.degree. C., for a commercially reasonable period of time,
such as at least about one day, at least about one week, at least
about one month, at least about three months, at least about six
months, at least about one year, or at least about two years.
[0102] A formulation should suit the desired mode of
administration. The agents of use with the current disclosure may
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,
ophthalmic, buccal, and rectal. The individual agents may also be
administered in combination with one or more additional agents or
together with other biologically active or biologically inert
agents. Such biologically active or inert agents may be in fluid or
mechanical communication with the agent(s) or attached to the
agent(s) by ionic, covalent, Van der Waals, hydrophobic,
hydrophilic or other physical forces.
[0103] Controlled-release (or sustained-release) preparations may
be formulated to extend the activity of the agent(s) and reduce
dosage frequency. Controlled-release preparations may also be used
to affect the time of onset of action or other characteristics,
such as blood levels of the agent, and consequently affect the
occurrence of side effects. Controlled-release preparations may be
designed to initially release an amount of an agent(s) that
produces the desired therapeutic effect, and gradually and
continually release other amounts of the agent to maintain the
level of therapeutic effect over an extended period. In order to
maintain a near-constant level of an agent in the body, the agent
may be released from the dosage form at a rate that will replace
the amount of agent being metabolized or excreted from the body.
The controlled-release of an agent may be stimulated by various
inducers (e.g., change in pH, change in temperature, enzymes,
water, or other physiological conditions or molecules).
[0104] Agents or compositions described herein may also be used in
combination with other therapeutic modalities, as described further
below. Thus, in addition to the therapies described herein, one may
also provide to the subject other therapies known to be efficacious
for treatment of the disease, disorder, or condition.
Therapeutic Methods
[0105] Also provided herein is a process of treating a
mitochondria-associated disease, disorder, or condition in a
subject in need of administration of a therapeutically effective
amount of a trans-4-hydroxycyclohexyl 6-phenylhexanamide derivative
mitofusin activator to prevent or treat a mitochondria-associated
disease, disorder, or condition.
[0106] For example, the compositions and methods described herein
may be used as a primary therapy for Charcot-Marie-Tooth or as an
adjunctive therapy for Huntington's disease, Parkinson's disease,
Alzheimer's disease, or ALS to retard or reverse disease
progression.
[0107] As another example, the compositions and methods described
herein may be used for the prevention or treatment of
chemotherapy-induced peripheral neuropathy. For example as pre- and
post-therapy for individuals undergoing scheduled chemotherapy for
the treatment of cancer. Pre- and post-chemotherapy treatment with
trans-4-hydroxycyclohexyl 6-phenylhexanamide derivative mitofusin
activators may prevent, attenuate, and accelerate recovery from
chemotherapy-induced peripheral neuropathy. This therapy may
minimize sensory and motor neuron susceptibility to
chemotherapeutic agents and accelerate repair of
chemotherapy-induced neuronal damage by promoting mitochondrial
fitness and localization to areas of injury and regrowth.
[0108] As yet another example, the compositions and methods
described herein may be used for the treatment of a physical
injury. For example, as a primary therapy for any contusive or
laceration injury involving the spine or peripheral nerves (perhaps
even the brain, (i.e., concussion), such as motor vehicle or sports
injuries. This therapy may help restore normal motor function by
augmenting regeneration and repair of injured neurons.
[0109] Methods described herein are generally performed on a
subject in need thereof. A subject in need of the therapeutic
methods described herein may be a subject having, diagnosed with,
suspected of having, or at risk for developing a
mitochondria-associated disease, disorder, or condition. A
determination of the need for treatment will typically be assessed
by a history and physical exam consistent with the disease or
condition at issue. Diagnosis of the various conditions treatable
by the methods described herein is within the skill of the art. The
subject may be an animal subject, including a mammal, such as
horses, cows, dogs, cats, sheep, pigs, mice, rats, monkeys,
hamsters, guinea pigs, chickens, and humans. For example, the
subject may be a human subject.
[0110] Generally, a safe and effective amount of a
trans-4-hydroxycyclohexyl 6-phenylhexanamide derivative mitofusin
activator is, for example, that amount that would cause the desired
therapeutic effect in a subject while minimizing undesired side
effects. In various aspects, an effective amount of a mitofusin
activator described herein may substantially inhibit
mitochondria-associated disease, disorder, or condition, slow the
progress of mitochondria-associated disease, disorder, or
condition, or limit the development of mitochondria-associated
disease, disorder, or condition. For example, a desired therapeutic
effect may be a delay in peripheral neuropathy (e.g., over the
course of three years) compared to placebo assessed by slower
increase in modified composite CMT neuropathy score. As another
example, a desired therapeutic effect may be reversal or absence of
progression of peripheral neuropathy compared to placebo, as
indicated by lower or stable modified composite CMT neuropathy
score. As yet another example, a desired therapeutic effect may be
reversal or absence of progression of dysregulated motor function
or increased regeneration and repair of injured neurons.
[0111] According to the methods described herein, administration
may be parenteral, pulmonary, oral, topical, intradermal,
intramuscular, intraperitoneal, intravenous, subcutaneous,
intranasal, epidural, ophthalmic, buccal, or rectal
administration.
[0112] When used in the treatments described herein, a
therapeutically effective amount of a mitofusin activator may be
employed in pure form or, where such forms exist, in
pharmaceutically acceptable salt form and with or without a
pharmaceutically acceptable excipient. For example, the compounds
of the present disclosure may be administered, at a reasonable
benefit/risk ratio applicable to any medical treatment, in a
sufficient amount to treat, reverse, prevent, or slow the
progression of mitochondria-associated disease, disorder, or
condition.
[0113] The amount of a composition described herein that may be
combined with a pharmaceutically acceptable carrier to produce a
single dosage form will vary depending upon the host treated and
the particular mode of administration. It will be appreciated by
those skilled in the art that the unit content of agent contained
in an individual dose of each dosage form need not in itself
constitute a therapeutically effective amount, as the necessary
therapeutically effective amount could be reached by administration
of a number of individual doses.
[0114] Toxicity and therapeutic efficacy of compositions described
herein may be determined by standard pharmaceutical procedures in
cell cultures or experimental animals for determining the LD.sub.50
(the dose lethal to 50% of the population) and the ED.sub.50, (the
dose therapeutically effective in 50% of the population). The dose
ratio between toxic and therapeutic effects is the therapeutic
index that may be expressed as the ratio LD.sub.50/ED.sub.50, where
larger therapeutic indices are generally understood in the art to
be optimal.
[0115] The specific therapeutically effective dose level for any
particular subject will depend upon a variety of factors including
the disorder being treated and the severity of the disorder;
activity of the specific compound employed; the specific
composition employed; the age, body weight, general health, sex and
diet of the subject; the time of administration; the route of
administration; the rate of excretion of the composition employed;
the duration of the treatment; drugs used in combination or
coincidental with the specific compound employed; and like factors
well known in the medical arts (see 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.sup.th 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 doses of the composition at levels lower than those required
to achieve the desired therapeutic effect and to gradually increase
the dosage until the desired effect is achieved. If desired, the
effective daily dose may be divided into multiple doses for
purposes of administration. Consequently, single dose compositions
may contain such amounts or submultiples thereof to make up the
daily dose. It will be understood, however, that the total daily
usage of the compounds and compositions of the present disclosure
will be decided by an attending physician within the scope of sound
medical judgment.
[0116] Again, each of the states, diseases, disorders, and
conditions, described herein, as well as others, may benefit from
compositions and methods described herein. Generally, treating a
state, disease, disorder, or condition includes preventing or
delaying the appearance of clinical symptoms in a mammal that may
be afflicted with or predisposed to the state, disease, disorder,
or condition but does not yet experience or display clinical or
subclinical symptoms thereof. Treating may also include inhibiting
the state, disease, disorder, or condition (e.g., arresting or
reducing the development of the disease or at least one clinical or
subclinical symptom thereof). Furthermore, treating may include
relieving the disease (e.g., causing regression of the state,
disease, disorder, or condition or at least one of its clinical or
subclinical symptoms). A benefit to a subject to be treated may be
either statistically significant or at least perceptible to the
subject or to a physician.
[0117] Administration of a trans-4-hydroxycyclohexyl
6-phenylhexanamide derivative mitofusin activator may occur as a
single event or over a time course of treatment. For example, a
mitofusin activator may be administered daily, weekly, bi-weekly,
or monthly. For treatment of acute conditions, the time course of
treatment will usually be at least several days. Certain conditions
could extend treatment from several days to several weeks. For
example, treatment could extend over one week, two weeks, or three
weeks. For chronic conditions, treatment could extend from several
weeks to several months or even years.
[0118] Treatment in accord with the methods described herein may be
performed prior to, concurrent with, or after conventional
treatment modalities for treating, preventing, or slowing the
progression of mitochondria-associated disease, disorder, or
condition.
[0119] A trans-4-hydroxycyclohexyl 6-phenylhexanamide derivative
mitofusin activator may be administered simultaneously or
sequentially with another agent, such as an antibiotic, an
anti-inflammatory, or another neuroregenerative or neurotherapeutic
agent. For example, a trans-4-hydroxycyclohexyl 6-phenylhexanamide
derivative mitofusin activator may be administered simultaneously
with another agent, such as an antibiotic or an anti-inflammatory.
Simultaneous administration may occur through administration of
separate compositions, each containing one or more of a mitofusin
activator, an antibiotic, an anti-inflammatory, or another agent.
Simultaneous administration may occur through administration of one
composition containing two or more of a mitofusin activator, an
antibiotic, an anti-inflammatory, or another agent. A
trans-4-hydroxycyclohexyl 6-phenylhexanamide derivative mitofusin
activator may be administered sequentially with an antibiotic, an
anti-inflammatory, or another agent. For example, a
trans-4-hydroxycyclohexyl 6-phenylhexanamide derivative mitofusin
activator may be administered before or after administration of an
antibiotic, an anti-inflammatory, or another agent.
Administration
[0120] Agents and compositions described herein may be administered
according to methods described herein in a variety of means known
to the art. The agents and composition may be used therapeutically
either as exogenous materials or as endogenous materials. Exogenous
agents are those produced or manufactured outside of the body and
administered to the body. Endogenous agents are those produced or
manufactured inside the body by some type of device (biologic or
other) for delivery within or to other organs in the body.
[0121] As discussed above, administration may be parenteral,
pulmonary, oral, topical, transdermal (e.g., a transdermal patch)
intradermal, intramuscular, intraperitoneal, intravenous,
subcutaneous, intranasal, epidural, ophthalmic, buccal, or rectal
administration.
[0122] Agents and compositions described herein may be administered
in a variety of methods well known in the arts. Administration
methods may include, for example, methods involving oral ingestion,
direct injection (e.g., systemic or stereotactic), implantation of
cells engineered to secrete the factor of interest, drug-releasing
biomaterials, polymer 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 .mu.m), nanospheres
(e.g., less than 1 .mu.m), microspheres (e.g., 1-100 .mu.m),
reservoir devices, a combination of any of the above, or other
suitable delivery vehicles to provide the desired release profile
in varying proportions. Other methods of controlled-release
delivery of agents or compositions will be known to the skilled
artisan and are within the scope of the present disclosure.
[0123] Delivery systems may include, for example, an infusion pump
that may be used to administer the agent or composition in a manner
similar to that used for delivering insulin or chemotherapy to
specific organs or tumors. Typically, using such a system, an agent
or composition may be administered in combination with a
biodegradable, biocompatible polymeric 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. In addition, a controlled
release system may be placed in proximity of a therapeutic target,
thus requiring only a fraction of a systemic dosage.
[0124] Agents may be encapsulated and administered in a variety of
carrier delivery systems. Examples of carrier delivery systems
include microspheres, hydrogels, polymeric implants, smart
polymeric carriers, and liposomes (see generally, Uchegbu and
Schatzlein, eds. (2006) Polymers in Drug Delivery, CRC, ISBN-10:
0849325331). Carrier-based systems for molecular or biomolecular
agent delivery can: provide for intracellular delivery, tailor
biomolecule/agent release rates; increase the proportion of
biomolecule that reaches its site of action; improve the transport
of the drug to its site of action; allow colocalized deposition
with other agents or excipients; improve the stability of the agent
in vivo; prolong the residence time of the agent at its site of
action by reducing clearance; decrease the nonspecific delivery of
the agent to nontarget tissues; decrease irritation caused by the
agent; decrease toxicity due to high initial doses of the agent;
alter the immunogenicity of the agent; decrease dosage frequency,
improve taste of the product; or improve shelf life of the
product.
Kits
[0125] Also provided herein are kits. Such kits may include an
agent or composition described herein and, in certain aspects,
instructions for administration. Such kits may facilitate
performance of the methods described herein. When supplied as a
kit, the different components of the composition may be packaged in
separate containers and admixed immediately before use. Components
include, but are not limited to MFN1, MFN2, activator target
peptides, or trans-4-hydroxycyclohexyl 6-phenylhexanamide
derivative mitofusin activators. Such packaging of the components
separately can, if desired, be presented in a pack or dispenser
device, which may 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 packaging of the
components separately may also, in certain instances, permit
long-term storage without losing activity of the components.
[0126] Kits may also include reagents in separate containers (e.g.,
sterile water or saline) to be added to a lyophilized active
component packaged separately. For example, sealed glass ampules
may contain a lyophilized component and in a separate ampule,
sterile water, sterile saline or sterile each of which has been
packaged under a neutral non-reacting gas, such as nitrogen.
Ampules may consist of any suitable material, such as glass,
organic polymers, such as polycarbonate, polystyrene, ceramic,
metal or any other material typically employed to hold reagents.
Other examples of suitable containers include bottles that may be
fabricated from similar substances as ampules, and envelopes that
may consist of foil-lined interiors, such as aluminum or an alloy.
Other containers include test tubes, vials, flasks, bottles,
syringes, and the like. Containers may have a sterile access port,
such as a bottle having a stopper that may be pierced by a
hypodermic injection needle. Other containers may have two
compartments that are separated by a readily removable membrane
that upon removal permits the components to mix. Removable
membranes may be glass, plastic, rubber, and the like.
[0127] In certain aspects, kits may be supplied with instructional
materials. Instructions may be printed on paper or other substrate,
and/or may be supplied as an electronic-readable medium, such as a
floppy disc, mini-CD-ROM, CD-ROM, DVD-ROM, Zip disc, videotape,
audio tape, and the like. Detailed instructions may not be
physically associated with the kit; instead, a user may be directed
to an Internet web site specified by the manufacturer or
distributor of the kit.
[0128] Compositions and methods described herein utilizing
molecular biology protocols may be according to a variety of
standard techniques known to the art (see, e.g., Sambrook and
Russel (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 Russel (2001) Molecular Cloning: A Laboratory Manual, 3d ed.,
Cold Spring Harbor Laboratory Press, ISBN-10: 0879695773; Elhai, J.
and Wolk, C. P. 1988. Methods in Enzymology 167, 747-754; Studier
(2005) Protein 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).
Metabolites
[0129] Metabolites of
trans-(4-hydroxycyclohexyl)-6-phenylhexanamide were isolated
following in vivo administration and certain examples were found to
be highly active mitofusin activators. Among mitofusin activators
that are metabolites of
trans-(4-hydroxycyclohexyl)-6-phenylhexanamide include the
following compounds:
##STR00012##
[0130] Mitofusin activators having structures analogues to those
produced metabolically are also encompassed within the compounds
disclosed herein. Any of the foregoing mitofusin activator
metabolites may be administered to a subject directly for purposes
of managing a mitochondria-associated disease, disorder or
condition.
[0131] Definitions and methods described 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 according to
conventional usage by those of ordinary skill in the relevant
art.
[0132] In some aspects, numbers expressing quantities of
ingredients, properties such as molecular weight, reaction
conditions, and so forth, used to describe and claim certain
aspects of the present disclosure are to be understood as being
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 of the mean for the device or method being
employed to determine the value. In some features, the numerical
parameters set forth in the written description and attached claims
are approximations that may vary depending upon the desired
properties sought to be obtained by a 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 recited herein.
[0133] In some aspects, the terms "a" and "an" and "the" and
similar references used in the context of describing a particular
aspect (especially in the context of certain of the following
claims) may be construed to cover both the singular and the plural,
unless specifically noted otherwise. In some aspects, the term "or"
as used herein, including the claims, is used to mean "and/or"
unless explicitly indicated to refer to alternatives only or the
alternatives are mutually exclusive.
[0134] The terms "comprise," "have" and "include" are open-ended
linking verbs. Any forms or tenses of one or more of these verbs,
such as "comprises," "comprising," "has," "having," "includes" and
"including," are open-ended. For example, any method that
"comprises," "has" or "includes" one or more steps is not limited
to possessing only those one or more steps and may cover other
unlisted steps. Similarly, any composition or device that
"comprises," "has" or "includes" one or more features is not
limited to possessing only those one or more features and may cover
other unlisted features.
[0135] All methods described herein may be performed in any
suitable order unless otherwise indicated herein or otherwise
clearly contradicted by context. The use of any and all examples,
or exemplary language (e.g., "such as") provided with respect to
certain aspects herein is intended merely to better illuminate the
present disclosure and does not pose a limitation on the scope of
the present disclosure otherwise claimed. No language in the
specification should be construed as indicating any non-claimed
element essential to the practice of the present disclosure.
[0136] Groupings of alternative elements, embodiments, aspects, or
features of the present disclosure disclosed herein are not to be
construed as limitations. 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. One or more members of a
group may be included in, or deleted from, a group for reasons of
convenience or patentability. When any such inclusion or deletion
occurs, the specification is herein deemed to contain the group as
modified thus fulfilling the written description of all Markush
groups used in the appended claims.
[0137] Citation of a reference herein shall not be construed as an
admission that such is prior art to the present disclosure.
[0138] Having described the present disclosure in detail, it will
be apparent that modifications, variations, and equivalent
embodiments, features, or aspects are possible without departing
the scope of the present disclosure defined in the appended claims.
Furthermore, it should be appreciated that all examples in the
present disclosure are provided as non-limiting examples.
EXAMPLES
[0139] The following non-limiting examples are provided to further
illustrate the present disclosure. It should be appreciated by
those of skill in the art that the techniques disclosed in the
examples that follow represent approaches the inventors have found
function well in the practice of the present disclosure, and thus
may be considered to constitute examples of modes for its practice.
However, those of skill in the art should, in light of the present
disclosure, appreciate that many changes may be made in the
specific features that are disclosed and still obtain a like or
similar result without departing from the spirit and scope of the
present disclosure.
Example 1. N-(cis-4-Hydroxycyclohexyl)-6-phenylhexanamide (MiM 111
Cis, Compound 15A)
##STR00013##
[0141] To a solution of 6-phenylhexanoic acid 1.2 (200 mg, 1.04
mmol, 196 .mu.L, 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
.mu.L, 2.00 eq.) in DMF (3 mL) was added HOBt (169 mg, 1.25 mmol,
1.20 eq.) and EDCl (299 mg, 1.56 mmol, 1.50 eq.). The mixture was
stirred at 10.degree. C. for 10 hours. The mixture was diluted with
water (20 mL) and extracted with EtOAc (10 mL.times.3). The organic
phase was washed with 1M aqueous HCl (30 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by preparative HPLC.
The title compound was obtained as a light yellow gum. MS:
m/z=290.1 (M+H).sup.+; .sup.1H NMR (400 MHz): MeOD .delta.
7.25-7.22 (m, 2H), 7.17-7.11 (m, 3H), 3.84 (s, 1H), 3.70-3.65 (m,
1H), 2.60 (t, J=7.6 Hz, 2H), 2.16 (t, J=7.2 Hz, 2H), 1.73-1.65 (m,
3H), 1.65-1.59 (m, 9H), 1.36-1.34 (m, 2H). .sup.13C NMR (400 MHz):
MeOD .delta.: 175.628, 143.895, 129.579, 129.414, 126.809, 66.945,
37.182, 36.885, 32.558, 32.179, 29.863, 28.074, 27.168.
Example 2. N-(trans-4-Hydroxycyclohexyl)-6-phenylhexanamide (MiM
111 Trans, Compound 15B)
##STR00014##
[0143] To a solution of 6-phenylhexanoic acid 1.2 (200 mg, 1.04
mmol, 196 .mu.L, 1.00 eq.) and trans-4-hydroxycyclohexylamine 2.1
(174 mg, 1.14 mmol, 1.10 eq.) and DIEA (269 mg, 2.08 mmol, 362
.mu.L, 2.00 eq.) in DMF (3 mL) was added HOBt (169 mg, 1.25 mmol,
1.20 eq.) and EDCl (299 mg, 1.56 mmol, 1.50 eq.). The mixture was
stirred at 10.degree. C. for 10 hours. The mixture was diluted with
water (20 mL) and extracted with EtOAc (10 mL.times.3). The organic
phase was washed with 1M aqueous HCl (30 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by preparative HPLC.
The title compound was obtained as a white solid. MS: m/z=290.1
(M+H).sup.+; .sup.1H NMR (400 MHz): MeOD .delta. 7.25-7.21 (m, 2H),
7.16-7.11 (m, 3H), 3.59-3.48 (m, 1H), 2.60 (t, J=7.6 Hz, 2H), 2.13
(t, J=7.6 Hz, 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). .sup.13C NMR (400 MHz): MeOD .delta.:
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.
Example 3: The Trans-Stereoisomer of MiM 111 is an Allosteric
Mitofusin Agonist, Whereas the Cis-Stereoisomer of MiM 111 is
Inactive
[0144] Previously, chemical syntheses of candidate mitofusin
activators sourced starting materials as a mixture of
stereoisomers, resulting in products that were also mixtures of 2
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 are no
data describing, nor even computer models that can predict: 1. if
stereoisomers affect mitofusin activator efficacy; and 2. if they
do, which stereoisomer(s) have superior properties. Accordingly,
cis- and trans-diastereomers of
N-(4-hydroxycyclohexyl)-6-phenylhexanamide, designated cis- and
trans-MiM 111, were individually synthesized, validated, and
characterized (FIGS. 3A-3B). Remarkably, the cis form exhibited no
detectable mitofusin activating activity, measured as mitochondrial
elongation, in either MFN1 or MFN2 deficient murine embryonic
fibroblasts (FIGS. 2A-2B). By contrast, the trans form of MiM 111
was equipotent to a prototype mitofusin agonist of the chemical
class described in Rocha, et al.; Science 2018, Chimera C.
Functional activity of cis- and
trans-N-(4-hydroxycyclohexyl)-6-phenylhexanamide mirrored their
ability to provoke the characteristic change in MFN2 conformation
that underlies its activation (FIG. 2C), mechanistically linking
biological effect and target engagement as a function of isomeric
structure. The cis- and trans-stereoisomers of MiM 111 had similar
in vitro pharmacokinetic profiles (Table 1).
TABLE-US-00001 TABLE 1 In vitro functional and pharmacokinetic
properties of MiM 111 cis- and trans- diastereomers (Compounds 15A
and 15B) EC.sub.50 EC.sub.50 (nM) (nM) t.sub.1/2 t.sub.1/2 PAMPA
Mfn1 Mfn2 hPPB mPPB HLM MLM Pe Cpd Structure KO KO (%) (%) (min)
(min) (nm/sec) 15A ##STR00015## inactive inactive 93.9 93.8 102 70
42.6 15B ##STR00016## 7.7 9.5 90.4 96.7 >145 127 22.9
Example 4: Trans-Stereoisomers of MiM 111 Having Oxy-Substituted
Linkers are Potent Mitofusin Activators with a Spectrum of Passive
Membrane Permeability Characteristics
[0145] The complete series of oxy-substituted linker analogs for
trans-N-(4-hydroxycyclohexyl)-6-phenylhexanamide were synthesized
and evaluated for fusogenic activity and pharmacokinetic
properties. All of the oxy-substituted trans-analogs retained
excellent fusogenic activity, but the position of the oxygen within
the linker altered the passive membrane permeability
characteristics of these compounds, with the carbamate exhibiting
10-fold greater PAMPA, but reduced microsomal stability, compared
to parent trans-N-(4-hydroxycyclohexyl)-6-phenylhexanamide (Table
2).
TABLE-US-00002 TABLE 2 Functional and pharmacokinetic properties of
oxy-substituted linker analogs of trans-MiM 111 ((Compound 15B)
t.sub.1/2 t.sub.1/2 PAMPA EC.sub.50 hPPB mPPB HLM MLM Pe Cpd
Structure (nM) (%) (%) (min) (min) (nm/sec) 15B ##STR00017## 5.54
90.4 96.7 >145 127 22.9 21 ##STR00018## 9.63 80.2 81.8 >145
>145 3.66 22 ##STR00019## 12.92 57.1 60.4 >145 >145 1.176
23 ##STR00020## 3.31 38.2 48.5 >145 >145 2.228 24
##STR00021## 7 31 55.2 66.4 >145 >145 23.799 25 ##STR00022##
6.28 93.3 94.8 131.3 48.9 210.149
[0146] Compounds 21-25 were synthesized similarly to Compound 15B
(Example 2), except incorporating an appropriate oxygenated
6-phenylhexanoic acid derivative for 6-phenylhexanoic acid 1.2.
Namely, Compounds 21-25 were synthesized by replacing
6-phenylhexanoic acid 1.2 with 5-phenoxypentanoic acid,
4-(benzyloxy)butanoic acid, 3-phenethoxypropanoic acid,
2-(3-phenylpropoxy)acetic acid, or 4-phenylbutylcaronochloridate,
respectively.
[0147] Compound 21: HPLC: RT: 2.17 min, purity: 99.9%. LC-MS:
RT=0.830 min, m/z=292.3 (M+H).sup.+. .sup.1H NMR: 400 MHz, MeOD.
.delta. 7.25 (t, J=8.0 16.4 Hz, 2H), 6.9 (d, J=8.4 Hz, 3H), 3.9 (m,
2H), 3.51-3.63 (m, 2H), 2.24 (s, 2H), 1.92-1.97 (m, 4H), 1.77-1.70
(m, 4H), 1.30-1.37 (m, 4H). .sup.13C NMR: (400 MHz, MeOD) .delta.
(176, 161, 131, 122, 116, 71.2, 69.2, 37.6, 35.6, 32.3, 30.7, 24.5)
ppm.
[0148] Compound 22: HPLC: RT: 1.80 min, purity: 99.8%. LC-MS:
RT=0.802 min, m/z=292.3 (2M+Na).sup.+. .sup.1H NMR: (400 MHz, MeOD)
.delta. 7.25-7.33 (m, 5H), 4.48 (s, 2H), 3.46-3.59 (m, 4H), 2.24
(t, J=7.2, 2H), 1.82-1.92 (m, 6H), 1.20-1.33 (m, 4H). .sup.13C NMR:
(400 MHz, MeOD) .delta. (175, 139, 130, 129, 128, 74.0, 70.6, 70.5,
34.9, 34.1, 31.6, 27.2) ppm.
[0149] Compound 23: HPLC: RT: 1.86 min, purity: 97.6%. LC-MS:
RT=0.789 min, m/z=292.4 (M+H).sup.+. .sup.1H NMR: (400 MHz, MeOD)
.delta. 7.17-7.27 (m, 5H), 3.66-3.70 (m, 4H), 3.49-3.64 (m, 2H),
2.84 (t, J=6.8, 2H), 2.37 (t, J=6.0, 2H), 1.82-1.94 (m, 4H),
1.17-1.34 (m, 4H). .sup.13C NMR: (400 MHz, MeOD) .delta. (173, 140,
130, 129, 127, 73.0, 70.5, 68.1, 37.9, 37.2, 34.9, 31.5) ppm.
[0150] Compound 24: HPLC: RT: 2.24 min, purity: 98.3%. LC-MS:
RT=0.825 min, m/z=292.1 (M+H).sup.+. .sup.1H NMR: (400 MHz, MeOD)
.delta. 7.24-7.28 (m, 2H), 7.15-7.20 (m, 3H), 3.88 (s, 2H),
3.68-3.70 (m, 1H), 3.53-3.56 (m, 1H), 3.50 (t, J=6.4 Hz, 2H), 2.70
(t, J=7.6 Hz, 2H), 1.91-1.96 (m, 4H), 1.86-1.87 (m, 2H), 1.33-1.38
(m, 4H). .sup.13C NMR: (400 MHz, MeOD) .delta. (172, 143, 130, 129,
127, 72.1, 71.1, 70.4, 34.9, 33.3, 32.3, 31.4) ppm.
[0151] Compound 25: HPLC: RT: 2.54 min, purity: 99.7%. LC-MS:
RT=0.886 min, m/z=292.2 (M+H).sup.+. .sup.1H NMR: (400 MHz, MeOD)
.delta. 7.12-7.26 (m, 5H), 4.02 (t, J=6.0, 2H), 3.47-3.52 (m, 1H),
3.33-3.35 (m, 1H), 2.63 (t, J=7.2, 2H), 1.88-1.94 (m, 4H),
1.64-1.66 (m, 4H), 1.1.23-1.31 (m, 4H). .sup.13C NMR: (400 MHz,
MeOD) .delta. (158, 143, 130, 129, 126, 70.5, 65.6, 50.7, 36.5,
34.9, 31.9, 29.9, 29.1) ppm.
Example 4B: The Cyclic Cyclopropane Backbone Analogue of
Trans-Stereoisomer MiM 111 is Also Active
[0152] Compound 26 was synthesized similarly to Compound 15B
(Example 2), except incorporating
2-(3-phenylpropyl)cyclopropane-1-carboxylic acid for
6-phenylhexanoic acid 1.2. This compound had an EC.sub.50 of 5.1
nm, H and M plasma protein binding of 94.4% and 95.5%, H and M
liver microsome stability T.sup.1/2 values of >145 minutes and
114.1 minutes, and a PAMPA assay value of 58.451 nm/s.
Example 5: In Vivo Pharmacological Profiling Shows that Trans-MiM
111 (Compound 15B) is a Clinical Candidate for CMT2A
[0153] Trans-MiM 111 exhibited high microsomal stability and
passive permeability (PAMPA Pe), with a low efflux ratio in NIH
MDR1 cells that did not change 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 anticipated that microsomal
stability would correlate with in vivo plasma t.sup.1/2, and
passive permeability with CNS levels.
TABLE-US-00003 TABLE 3 P-gp studies of trans-MiM 111 (Compound 15B)
Mean P.sub.app Mean Compound GF120918 (10.sup.-6 cm/s) Efflux
Recovery % ID (+/-) A to B B to A Ratio A to B B to A Note Nadolol
- 0.26 ND ND 97.88 ND Low permeability marker Metoprolol - 21.24 ND
ND 105.27 ND High permeability marker Digoxin - 0.11 13.36 123.06
98.93 103.29 P-gp + 0.73 2.05 2.81 97.04 99.34 substrate 15B -
20.36 35.45 1.74 84.58 100.21 -- + 31.88 23.21 0.73 84.76
100.61
[0154] This notion was examined by in vivo PK studies in mice, the
only species in which pre-clinical models of human CMT2A have been
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). A target therapeutic brain level of 10
times the in vitro EC.sub.50 for each compound, or 30 ng/g
(.about.100 nM), was established.
[0155] Comparative mouse plasma and brain levels were determined at
increasing times after a single 10 mg/kg intravenous (IV) dose. The
Vdss (volume of distribution at steady state) of trans-MiM 111 was
0.35 L/kg and peak brain levels were 2,793 ng/g; therapeutic levels
(of >30 ng/g) were maintained for more than 2 hours after the
single IV dose (FIG. 4A).
[0156] Because of its low Vdss, high Pe, and low efflux in NIH MDR1
cells it was surmised that trans-MiM 111 might accumulate in brains
over time. If this were so, then brain levels after single dose
administration could be misleading vis-a-vis therapeutic efficacy.
To test this idea osmotic mini-pumps were used to deliver trans-MiM
111 subcutaneously (SQ) at a daily dose of 60 mg/kg/day for three
days to achieve steady state, and its elimination kinetics were
defined after mini-pump removal (FIG. 4B). Trans-MiM 111 plasma
half-lives were similar after bolus IV and chronic SQ
administration (1.1 hours and 1.33 hours, respectively), but its
brain half-life was substantially longer after chronic infusion
(3.37 hours vs 1.06 hours after IV). Because it is 96.7% plasma
protein bound in mice, the unbound fraction (fu) of trans-MiM 111
is 0.033. Using this factor to convert plasma [Cpd 15B].sub.total
to plasma [Cpd 15B].sub.free/unbound, the calculated ratio of brain
to free plasma level is 10.8 after chronic infusion.
Example 6: In Vivo Trans-MiM 111 Engages its Mitochondrial Targets
in a Pre-Clinical Mouse Model of CMT2A
[0157] Taken together, the above results indicated that trans-MiM
111 has properties enabling it to activate mitofusins of neuronal
mitochondria in CMT2A mice in vivo. In the only published study a
prototype mitofusin activator, Chimera B-A/I, increased
mitochondrial motility when topically applied to sciatic nerve
neurons ex vivo (Rocha, A. G., et al.; Science 360:336-41, 2018).
The in vivo effects of mitofusin activators on CMT2A neuronal
mitochondria were never assessed because this chemical class of
mitofusin agonists has a very short in vivo plasma half-life of
.about.0.2 hour. Because trans-MiM 111 is >75% orally
bioavailable (FIG. 5A) its ability to engage peripheral nerve
mitochondrial targets in vivo was assessed after a single oral
dose. Transgenic mice expressing the human MFN2 T105M mutation in
motor neurons (Rocha A. G., et al.; Science 360:336-41, 2018)
received 50 mg/kg of trans-MiM 111 and mitochondrial motility in
sciatic nerve neurons was assessed by a blinded investigator 6
hours thereafter. trans-MiM 111 markedly increased both the number
and velocity of motile mitochondria in CMT2A mouse neurons (FIG.
5B).
Example 7: Off-Target, Specificity, and Safety Studies Show that
Trans-MiM 111 has Properties of an Advanced Clinical Lead for
CMT2A
[0158] Off-target, specificity, and safety studies performed to
determine the suitability of trans-MiM 111 for possible clinical
translation (Table 4) demonstrated that it is a potent and
selective mitofusin activator with a favorable drug profile. It
exhibited sub-10 nM potency for both MFN1 and MFN2, with very low
inhibitory activity for cytochrome P450 enzymes, indicating a small
likelihood for drug-drug interactions. Activity screening against
hERG, hNAV1.5, hKCNQ, and a panel of 42 receptors/kinases revealed
only mild inhibition of dopamine amino transferase (DAT) and
monoamine oxidase (MAO-A; .about.30% inhibition at 10 .mu.M),
indicating a safety window of 1,000-fold compared to on-target
efficacy, and a correspondingly limited potential for off-target
side-effects.
TABLE-US-00004 TABLE 4 Summary of key properties of trans-MiM 111
(Compound 158) On-target potency EC.sub.50 Mfn1 KO cells (nm) 7.7
EC.sub.50 Mfn2 KO cells (nm) 9.5 Physical properties MW/cLogP/TPSA
289/3.22/49.33 .ANG. kinetic solubility (mM) 175 fsp3 (%) 61
Selectivity and safety profiles hNav1.5 (% inhib @ 10 mM) 8.3 hKCNQ
(% inhib @ 10 mM) 0 hERG patch clamp IC.sub.50 (mM) >30 44
receptor/kinase panel (10 mM) DAT and MAO-A > 30% inhib
CYP1A2/209/2019/2D6/3A4-M >50/>50/>50/>50/>50
IC.sub.50 (mM) AMES test negative
Materials and Methods
Cell Lines
[0159] Wild-type MEFs were prepared from E10.5 c57/bl6 mouse
embryos. SV-40 T antigen-immortalized MFN1 null (CRL-2992), MFN2
null (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, 1.times. nonessential amino
acids, 2 mM L-glutamine, 100 units/mL penicillin and 100 .mu.g/mL
streptomycin.
Confocal Live Cell Studies of Mitochondria
[0160] Live cell imaging was performed on an Olympus Diaphot 200
fluorescence microscope equipped with a 60.times. water immersion
objective. All live cells were grown on coated glass-bottom 12-well
plates and studied in modified Krebs-Henseleit buffer (138 mM NaCl,
3.7 mM KCl, 1.2 mM KH.sub.2PO.sub.4, 15 mM, 20 mM HEPES and 1 mM
CaCl.sub.2) at room temperature.
[0161] Cells were excited with 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, mitochondrial aspect
ratio (long axis/short axis) was calculated using automated edge
detection and Image J software. Mitochondrial depolarization was
calculated as percent of green mitochondria visualized on
MitoTracker Green and TMRE merged images, expressed as
green/(green+yellow mitochondria).times.100.
Preparative HPLC
[0162] Purification was performed using HPLC (H.sub.2O-MeOH;
Agilent 1260 Infinity systems equipped with DAD and mass-detectors.
Waters SunFire C18 OBD Prep Column, 100 .ANG., 5 .mu.m, 19
mm.times.100 mm with SunFire C18 Prep Guard Cartridge, 100 .ANG.,
10 .mu.m, 19 mm.times.10 mm) was used for separation. The material
was dissolved in 0.7 mL DMSO. Flow rate: 30 mL/minute. Purity of
the obtained fractions was checked via analytical LCMS. Spectra
were recorded for each fraction as it was obtained straight after
chromatography in the solution form. The solvent was evaporated in
the flow of N.sub.2 at 80.degree. C. On the basis of
post-chromatography LCMS analysis, fractions were combined united.
Solid fractions were dissolved in 0.5 mL MeOH and transferred into
pre-weighted marked vials.
Obtained solutions were again evaporated in the flow of N.sub.2 at
80.degree. C. After drying, products were characterized by LCMS,
.sup.1H NMR, and .sup.13C NMR.
HPLC/HRMS (ESI)
[0163] LC/MS analysis was carried out using Agilent 1100 Series
LC/MSD system with DAD\ELSD and Agilent LC\MSD VL (G1956 .ANG.), SL
(G1956B) mass-spectrometer or Agilent 1200 Series LC/MSD system
with DAD\ELSD and Agilent LC\MSD SL (G6130 .ANG.), SL (G6140 .ANG.)
mass-spectrometer. All the LC/MS data were obtained using
positive/negative mode switching. The compounds were separated
using a Zorbax SB-C18 1.8 .mu.m 4.6.times.15 mm Rapid Resolution
cartridge (PN 821975-932) under a mobile phase (A--ACN, 0.1% formic
acid; B--water (0.1% formic acid)). Flow rate: 3 mL/minute;
Gradient 0 minutes--100% B; 0.01 minute--100% B; 1.5 minutes--0% B;
1.8 minutes--0% B; 1.81 minutes--100% B; Injection volume 1 .mu.L;
Ionization mode atmospheric pressure chemical ionization (APCI);
Scan range m/z 80-1000.
Statistical Methods
[0164] Time-course and dose-response data are calculated for each
study using GraphPad Prism. All data are reported as mean.+-.SEM.
Statistical comparisons (two-sided) used one-way ANOVA and Tukey's
tests for multiple groups or Student's t-test for paired
comparisons. p<0.05 was considered significant. In vitro
pharmacokinetic analyses of mitofusin activators was performed at
WuXi Apptec Co. Ltd.
[0165] Binding to human and CD-1 mouse plasma proteins was measured
using equilibrium dialysis. Pooled individual frozen EDTA
anticoagulated plasma mouse and human samples were used as test
matrix. Warfarin was used as a positive control. The test compounds
were spiked into blank matrix at the final concentration of 2
.mu.M. A 150-.mu.L aliquot of matrix sample was added to one side
of the chamber in a 96-well equilibrium dialyzer plate (HTD
dialysis) and an equal volume of dialysis buffer was added to the
other side of the chamber. An aliquot of matrix sample was
harvested before the incubation and used as T.sub.0 samples for
recovery calculation. The incubations were performed in triplicate.
The dialyzer plate was placed in a humidified incubator and rotated
slowly for four hours at 37.degree. C. After incubation, the
samples were taken from the matrix side as well as the buffer side.
The plasma sample was matched with equal volume of blank buffer;
and buffer samples were matched with equal volume of blank plasma.
The matrix-matched samples were quenched with stop solution
containing internal standard. All samples were analyzed by
LC-MS/MS. All test compound concentrations in matrix and buffer
samples are expressed as peak area ratios (PAR) of analyte/internal
standard.
[0166] In vitro stability was measured in human and mouse liver
microsomes. An intermediate solution (100 .mu.M of small molecule)
was initially 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 a compound working solution or control working
solution was added to all wells of a 96-well plate for the time
points (minutes): T.sub.0, T.sub.5, T.sub.10, T.sub.20, T.sub.30,
T.sub.60, NCF60, except the matrix blank. The microsome solution
(680 .mu.L/well) (#452117, Corning; Woburn, Mass., USA; #R1000,
Xenotech; Kansas City, Kans., USA and #M1000, Xenotech; Kansas
City, Kans., USA) was dispersed to 96-well plate as reservoir
according to the plate map. Then, 80 .mu.L/well was added to every
plate by ADDA (Apricot Design Dual Arm, Apricot Designs, Inc.,
Covina, Calif., USA), and the mixture of microsome solution and
compound were allowed to incubate at 37.degree. C. for about 10
minutes. Next, 10 .mu.L of 100 mM potassium phosphate buffer/well
was added to NCF60 and incubated at 37.degree. C. (timer 1H was
started). After pre-warming, 90 .mu.L/well of NADPH (#00616, Sigma,
Aldrich, St. Louis, Mo., USA) regenerating system was dispensed to
96-well plate as reservoir according to the plate map. Then 10
.mu.L/well was added to every plate by ADDA to start reaction. To
terminate the reaction, 300 .mu.L/well of stop solution (cold in
4.degree. C., including 100 ng/mL tolbutamide and 100 ng/mL
labetalol as internal standards) was used, and sampling plates were
agitated for approximately 10 minutes. The samples were next
centrifuged at 4000 rpm for 20 minutes at 4.degree. C. Supernatants
were analyzed by LC-MS/MS.
Parallel Artificial Membrane Permeability Assay (PAMPA)
[0167] A 10 .mu.M solution of a small molecule in 5% DMSO (150
.mu.L) was added to each well of the donor plate, whose PVDF
membrane was pre-coated with 5 .mu.L of 1% brain polar lipid
extract (porcine)/dodecane mixture. Then, 300 .mu.L of PBS was
added to each well of the PTFE acceptor plate. The donor plate and
acceptor plate were combined together and incubated for 4 hours at
room temperature with shaking at 300 rpm. To prepare the T.sub.0
sample, 20 .mu.L of a donor solution was transferred to new well,
followed by the addition of 250 .mu.L PBS (DF:13.5) and 130 .mu.L
of ACN (containing internal standard) as the T.sub.0 sample. To
prepare the acceptor sample, the plate was removed from incubator
and 270 .mu.L of the solution was transferred from each acceptor
well and mixed with 130 .mu.L ACN (containing internal standard) as
an acceptor sample. To prepare the donor sample, 20 .mu.L of the
solution was transferred from each donor well and mixed with 250
.mu.L PBS (DF: 13.5), 130 .mu.L ACN (containing internal standard)
as a donor sample. The acceptor samples and donor samples were
analyzed by LC-MS/MS.
[0168] The present invention is also directed to the following
clauses.
[0169] Clause 1: A method of treating a peripheral nervous system
(PNS) or central nervous system (CNS) genetic disorder, physical
damage, and/or chemical injury, comprising: administering to a
subject a therapeutically effective amount of a composition
comprising one or more of a trans-stereoisomer 6-phenylhexanamide
derivative mitofusin activator or a pharmaceutically acceptable
salt thereof, wherein the trans-stereoisomer 6-phenylhexanamide
derivative mitofusin activator stimulates mitochondrial fusion,
increases mitochondrial fitness, and enhances mitochondrial
subcellular transport.
[0170] Clause 2. The method of clause 1, wherein the composition
comprises one or more mitofusin activators, wherein the mitofusin
activator comprises a structure of formula:
##STR00023##
[0171] or a pharmaceutically acceptable salt, tautomer, or
stereoisomer thereof,
[0172] wherein R.sup.1 is a non-, mono-, or poly-substituted
C.sub.3-8 cycloalkyl, C.sub.3-8 heteroaryl, C.sub.3-8 aryl, or
C.sub.3-8 heterocyclyl.
[0173] Clause 3: The method of any of clauses 1 to 2, wherein the
mitofusin activator comprises a structure of formula:
##STR00024##
[0174] and wherein R.sup.1 is
##STR00025##
[0175] Clause 4. The method of any of clauses 1 to 3, wherein
R.sup.1 is independently and optionally substituted by one or more
of acetamide, C.sub.1-8 alkoxy, amino, azo, Br, C.sub.1-8 alkyl,
carbonyl, carboxyl, Cl, cyano, C.sub.3-8 cycloalkyl, C.sub.3-8
heteroaryl, C.sub.3-8 heterocyclyl, hydroxyl, F, halo, indole, N,
nitrile, O, phenyl, S, sulfoxide, sulfone, and/or thiophene;
wherein R.sup.1 is optionally further substituted with one or more
acetamide, alkoxy, amino, azo, Br, C.sub.1-8 alkyl, carbonyl,
carboxyl, Cl, cyano, C.sub.3-8 cycloalkyl, C.sub.3-8 heteroaryl,
C.sub.3-8 heterocyclyl, hydroxyl, F, halo, indole, N, nitrile, O,
phenyl, S, sulfone, sulfur dioxide, and/or thiophene; and wherein
one or more of the alkyl, cycloalkyl, heteroaryl, heterocyclyl,
indole, or phenyl substituent is optionally further substituted
with one or more of the following substituents: acetamide, alkoxy,
amino, azo, Br, C.sub.1-8 alkyl, carbonyl, carboxyl, Cl, cyano,
C.sub.3-8 cycloalkyl, C.sub.3-8 heteroaryl, C.sub.3-8 heterocyclyl,
hydroxyl, F, halo, indole, N, nitrile, O, phenyl, S, sulfoxide,
sulfone, and thiophene.
[0176] Clause 5. The method of any of clauses 1 to 4, wherein the
mitofusin activator is:
##STR00026##
[0177] Clause 6. A compound of Formula II
##STR00027##
[0178] or a pharmaceutically acceptable salt thereof, wherein
[0179] Z is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;
[0180] R.sup.2 and R.sup.3 are independently selected from H, F,
alkyl, and C.sub.3-7 cycloalkyl, or R.sup.2 and R.sup.3 are taken
together to form a C.sub.3-7 cycloalkyl or heterocycloalkyl;
[0181] R.sup.4 and R.sup.5 are independently selected from H, F,
alkyl, and C.sub.3-7 cycloalkyl or R.sup.4 and R.sup.5 are taken
together to form a C.sub.3-7 cycloalkyl or heterocycloalkyl;
[0182] Y is O, CR.sup.6R.sup.7, CR.sup.8.dbd.CR.sup.9, a triple
bond, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, NR.sup.8, S,
SO.sub.2, SONR.sup.9, --NR.sup.9SO.sub.2--, --NR.sup.8CO--,
--CONR.sup.8--, or --NR.sup.8CONR.sup.9--;
[0183] R.sup.6 is H, F, alkyl, or cycloalkyl; and R.sup.7 is H, F,
alkyl, or cycloalkyl; or R.sup.6 and R.sup.7 are taken together to
form C.sub.3-7 cycloalkyl or heterocycloalkyl;
[0184] R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl;
[0185] R.sup.9 is H, alkyl, or C.sub.3-7 cycloalkyl;
[0186] o is 0, 1, 2, 3, 4 or 5;
[0187] p is 0 or 1; and
[0188] q is 0, 1, 2, 3, 4 or 5, wherein when o is equal to or
greater than 1, then Y=NR.sup.8, S, SO.sub.2, SONR.sup.9,
--NR.sup.9SO.sub.2--, --NR.sup.8CO--, --CONR.sup.8--,
--NR.sup.8CONR.sup.9--, and the sum of o+p+q is not less than 3 or
greater than 7.
[0189] Clause 7. The compound of clause 6, or a pharmaceutically
acceptable salt thereof, wherein
[0190] Z is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;
[0191] Y is O, CR.sup.6R.sup.7, cycloalkyl, or aryl;
[0192] R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7 and
R.sup.8 are independently selected from H and alkyl;
[0193] o is 0, 1, 2, 3, 4 or 5;
[0194] p is 0 or 1; and
[0195] q is 0, 1, 2, 3, 4 or 5;
[0196] wherein when o is equal to or greater than 1, then X is S or
SO.sub.2; and
[0197] wherein the sum of o+p+q is not less than 3 or greater than
7.
[0198] Clause 8. The compound of any of clauses 6 to 7, or a
pharmaceutically acceptable salt thereof, wherein
[0199] Z is aryl or heteroaryl;
[0200] Y is O, CH.sub.2, or cycloalkyl;
[0201] R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are CH.sub.2;
[0202] R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl;
[0203] R.sup.9 is or H, alkyl, and C.sub.3-7 cycloalkyl;
[0204] o is 0, 1, 2, 3, 4 or 5;
[0205] p is 0 or 1; and
[0206] q is 0, 1, 2, 3, 4 or 5;
[0207] wherein when o is equal to or greater than 1, then X is S or
SO.sub.2; and
[0208] wherein the sum of o+p+q is not less than 3 or greater than
5.
[0209] Clause 9. The compound of any of clauses 6 to 8, or a
pharmaceutically acceptable salt thereof, wherein
[0210] Z is aryl or heteroaryl;
[0211] Y is cyclopropyl or cyclobutyl;
[0212] R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are CH.sub.2;
[0213] R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl;
[0214] R.sup.9 is H, alkyl, COR.sup.7, or C.sub.3-7 cycloalkyl;
[0215] or R.sup.8 and R.sup.9 are taken together to form C.sub.3-7
cycloalkyl;
[0216] o is 0, 1, 2, or 3;
[0217] p is 1; and
[0218] q is 0, 1, 2, or 3;
[0219] wherein the sum of o+p+q is not less than 3 or greater than
5.
[0220] Clause 10. The compound of any of clauses 6 to 9, or a
pharmaceutically acceptable salt thereof, wherein
[0221] Z is aryl or heteroaryl;
[0222] Y is O or CH.sub.2;
[0223] R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are CH.sub.2;
[0224] R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl; R.sup.9 is H,
alkyl, COR.sup.7 or C.sub.3-7 cycloalkyl; or
[0225] R.sup.8 and R.sup.9 are taken together to form C.sub.3-7
cycloalkyl;
[0226] o is 0, 1, 2, 3 or 4;
[0227] p is 1; and
[0228] q is 0, 1, 2, 3 or 4;
[0229] wherein the sum of o+p+q is 5.
[0230] Clause 11. The compound of any of clauses 6 to 10, or a
pharmaceutically acceptable salt thereof, wherein
[0231] Z is aryl or heteroaryl;
[0232] Y is O or CH.sub.2;
[0233] R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are CH.sub.2;
[0234] o is 0, 1, 2, 3 or 4;
[0235] p is 1; and
[0236] q is 0, 1, 2, 3 or 4;
[0237] wherein the sum of o+p+q is 5.
[0238] Clause 12. The compound of any of clauses 6 to 11, or a
pharmaceutically acceptable salt thereof, wherein
[0239] Z is phenyl or heteroaryl; wherein the heterocyclic moiety
contains 1 to 4 atoms independently selected from nitrogen, oxygen
and sulfur, and wherein the phenyl or heterocyclic moiety has 0 to
4 substituents independently selected from R.sup.8, OR.sup.8, Cl,
F, --CN, CF.sub.3, --NR.sup.8R.sup.9, --SO.sub.2NR.sup.8R.sup.9,
--NR.sup.8SO.sub.2R.sup.9, --SO.sub.2R.sup.9, --CONR.sup.8R.sup.10,
--NR.sup.8COR.sup.10, C.sub.3-7 cycloalkyl, and
heterocycloalkyl;
[0240] Y is O or CH.sub.2;
[0241] R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are CH.sub.2;
[0242] R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl; R.sup.9 is H,
alkyl, COR.sup.7 or C.sub.3-7 cycloalkyl; or R.sup.8 and R.sup.9
are taken together to form C.sub.3-7 cycloalkyl;
[0243] R.sup.10 is independently selected from alkyl or C.sub.3-7
cycloalkyl;
[0244] o is 0, 1, 2, 3 or 4;
[0245] p is 1; and
[0246] q is 0, 1, 2, 3 or 4;
[0247] wherein the sum of o+p+q is 5.
[0248] Clause 13. The compound of any of clauses 6 to 12, or a
pharmaceutically acceptable salt thereof, wherein
[0249] Z is phenyl or heteroaryl; wherein the heterocyclic moiety
contains 1 to 3 atoms independently selected from nitrogen, oxygen
and sulfur, and wherein the phenyl or heterocyclic moiety has 0 to
3 substituents independently selected from R.sup.8, OR.sup.8, Cl,
F, --CN, CF.sub.3, --NR.sup.8R.sup.9, --SO.sub.2R.sup.9,
--CONR.sup.8R.sup.9, --NR.sup.7COR.sup.10, C.sub.3-7 cycloalkyl,
and heterocycloalkyl;
[0250] Y is O or CH.sub.2;
[0251] R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are CH.sub.2;
[0252] R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl; R.sup.9 is H,
alkyl, COR.sup.7 or C.sub.3-7 cycloalkyl; or R.sup.8 and R.sup.9
are taken together to form C.sub.3-7 cycloalkyl;
[0253] R.sup.10 is alkyl or C.sub.3-7 cycloalkyl;
[0254] o is 0, 1, 2, 3 or 4;
[0255] p is 1; and
[0256] q is 0, 1, 2, 3 or 4;
[0257] wherein the sum of o+p+q is 5.
[0258] Clause 14. The compound of any of clauses 6 to 13, or a
pharmaceutically acceptable salt thereof, wherein
[0259] Z is phenyl, 2-pyridinyl, 3-pyridinyl, 4-pyridinyl,
6-pyrimidinyl, 5-pyrimidinyl, 4-pyrimidinyl or 2-pyrimidinyl,
wherein the phenyl, pyridinyl, and pyrimidinyl moiety has 0 to 2
substituents independently selected from R.sup.8, OR.sup.8, Cl, F,
--CN, CF.sub.3, --NR.sup.8R.sup.9, --SO.sub.2R.sup.10,
--CONR.sup.8R.sup.9, and --NR.sup.8COR.sup.10
[0260] Y is O or CH.sub.2;
[0261] R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are CH.sub.2;
[0262] R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl;
[0263] R.sup.9 is H, alkyl, COR.sup.7 or C.sub.3-7 cycloalkyl;
or
[0264] R.sup.8 and R.sup.9 are taken together to form C.sub.3-7
cycloalkyl;
[0265] R.sup.10 is alkyl or C.sub.3-7 cycloalkyl;
[0266] o is 0, 1, 2, 3 or 4;
[0267] p is 1; and
[0268] q is 0, 1, 2, 3 or 4;
[0269] wherein the sum of o+p+q is 5.
[0270] Clause 15. The compound of any of clauses 6 to 14, or a
pharmaceutically acceptable salt thereof, wherein
[0271] Z is phenyl, 2-pyridinyl, 3-pyridinyl, 4-pyridinyl,
6-pyrimidinyl, 5-pyrimidinyl, 4-pyrimidinyl or 2-pyrimidinyl,
wherein the phenyl, pyridinyl, and pyrimidinyl moiety has 0 to 2
substituents independently selected from R.sup.8, OR.sup.8, Cl, F,
--CN, CF.sub.3, --NR.sup.8R.sup.9, --SO.sub.2R.sup.10,
--CONR.sup.8R.sup.9, and --NR.sup.8COR.sup.10
[0272] Y is O or CH.sub.2;
[0273] R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are CH.sub.2;
[0274] R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl; R.sup.9 is H,
alkyl, COR.sup.7 or C.sub.3-7 cycloalkyl; or
[0275] R.sup.8 and R.sup.9 are taken together to form C.sub.3-7
cycloalkyl;
[0276] R.sup.10 is alkyl or C.sub.3-7 cycloalkyl;
[0277] o is 0, 1, 2, 3 or 4;
[0278] p is 1; and
[0279] q is 0, 1, 2, 3 or 4;
[0280] wherein the sum of o+p+q is 5.
[0281] Clause 16. The compound of any of clauses 6 to 15, or
pharmaceutically acceptable salt thereof, wherein the compound
is
##STR00028##
[0282] Clause 17. The compound of any of clauses 6 to 15, or
pharmaceutically acceptable salt thereof, wherein the compound
is:
##STR00029##
[0283] Clause 18. A method of treating a disease for which a
mitofusin activator is indicated, the method comprising
administering to a mammal in need thereof a therapeutically
effective amount of a compound of Formula II
##STR00030##
[0284] or a pharmaceutically acceptable salt thereof, wherein
[0285] Z is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;
[0286] R.sup.2 and R.sup.3 are independently selected from H, F,
alkyl, and C.sub.3-7 cycloalkyl, or R.sup.2 and R.sup.3 are taken
together to form a C.sub.3-7 cycloalkyl or heterocycloalkyl;
[0287] R.sup.4 and R.sup.5 are independently selected from H, F,
alkyl, and C.sub.3-7 cycloalkyl or R.sup.4 and R.sup.5 are taken
together to form a C.sub.3-7 cycloalkyl or heterocycloalkyl;
[0288] Y is O, CR.sup.6R.sup.7, CR.sup.8.dbd.CR.sup.9, a triple
bond, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, NR.sup.8, S,
SO.sub.2, SONR.sup.9, --NR.sup.9SO.sub.2--, --NR.sup.8CO--,
--CONR.sup.8--, or --NR.sup.8CONR.sup.9--;
[0289] R.sup.6 is H, F, alkyl, or cycloalkyl; and R.sup.7 is H, F,
alkyl, or cycloalkyl; or R.sup.6 and R.sup.7 are taken together to
form C.sub.3-7 cycloalkyl or heterocycloalkyl;
[0290] R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl;
[0291] R.sup.9 is H, alkyl, or C.sub.3-7 cycloalkyl;
[0292] o is 0, 1, 2, 3, 4 or 5;
[0293] p is 0 or 1; and
[0294] q is 0, 1, 2, 3, 4 or 5, wherein when o is equal to or
greater than 1, then Y=NR.sup.8, S, SO.sub.2, SONR.sup.9,
--NR.sup.9SO.sub.2--, --NR.sup.8CO--, --CONR.sup.B--,
--NR.sup.8CONR.sup.9--, and the sum of o+p+q is not less than 3 or
greater than 7.
[0295] Clause 19. The method of any of clauses 1-5 or 18, wherein
the PNS or CNS disorder is selected from any one or a combination
of:
[0296] a chronic neurodegenerative condition wherein mitochondrial
fusion, fitness, or trafficking are impaired;
[0297] a disease or disorder associated with mitofusin 1 (MFN1) or
mitofusin 2 (MFN2) dysfunction;
[0298] a disease associated with mitochondrial fragmentation,
dysfunction, or dysmotility;
[0299] a degenerative neuromuscular condition such as
Charcot-Marie-Tooth disease, Amyotrophic Lateral Sclerosis,
Huntington's disease, Alzheimer's disease, Parkinson's disease;
[0300] hereditary motor and sensory neuropathy, autism, autosomal
dominant optic atrophy (ADOA), muscular dystrophy, Lou Gehrig's
disease, cancer, mitochondrial myopathy, diabetes mellitus and
deafness (DAD), Leber's hereditary optic neuropathy (LHON), Leigh
syndrome, subacute sclerosing encephalopathy, neuropathy, ataxia,
retinitis pigmentosa, and ptosis (NARP), myoneurogenic
gastrointestinal encephalopathy (MNGIE), myoclonic epilepsy with
ragged red fibers (MERRF), mitochondrial myopathy,
encephalomyopathy, lactic acidosis, stroke-like symptoms (MELAS),
mtDNA depletion, mitochondrial neurogastrointestinal
encephalomyopathy (MNGIE), dysautonomic mitochondrial myopathy,
mitochondrial channelopathy, or pyruvate dehydrogenase complex
deficiency (PDCD/PDH);
[0301] diabetic neuropathy;
[0302] chemotherapy-induced peripheral neuropathy; and/or
[0303] crush injury, spinal cord injury (SCI), traumatic brain
injury, stroke, optic nerve injury, and related conditions that
involve axonal disconnection.
[0304] Clause 20. The method of any of clauses 1-5, 18 or 19, with
the proviso that the mitofusin activator is not selected from the
following compounds:
##STR00031##
[0305] Clause 21. The method of any of clauses 1-5 or 18-20,
wherein the composition further comprises a pharmaceutically
acceptable excipient.
[0306] Clause 22. A method of treating a CNS or PNS genetic or
non-genetic neurodegenerative condition, injury, damage, or trauma
comprising administering to the subject a therapeutically effective
amount of the compound of any one of clauses 6 to 17.
[0307] Clause 23. The method of clause 22, wherein the subject is
diagnosed with or is suspected of having:
[0308] a chronic neurodegenerative condition wherein mitochondrial
fusion, fitness, or trafficking are impaired;
[0309] a disease or disorder associated with mitofusin 1 (MFN1) or
mitofusin 2 (MFN2) dysfunction;
[0310] a disease associated with mitochondrial fragmentation,
dysfunction, or dysmotility;
[0311] a degenerative neuromuscular condition such as
Charcot-Marie-Tooth disease, Amyotrophic Lateral Sclerosis,
Huntington's disease, Alzheimer's disease, Parkinson's disease;
[0312] hereditary motor and sensory neuropathy, autism, autosomal
dominant optic atrophy (ADOA), muscular dystrophy, Lou Gehrig's
disease, cancer, mitochondrial myopathy, diabetes mellitus and
deafness (DAD), Leber's hereditary optic neuropathy (LHON), Leigh
syndrome, subacute sclerosing encephalopathy, neuropathy, ataxia,
retinitis pigmentosa, and ptosis (NARP), myoneurogenic
gastrointestinal encephalopathy (MNGIE), myoclonic epilepsy with
ragged red fibers (MERRF), Mitochondrial myopathy,
encephalomyopathy, lactic acidosis, stroke-like symptoms (MELAS),
mtDNA depletion, mitochondrial neurogastrointestinal
encephalomyopathy (MNGIE), dysautonomic mitochondrial myopathy,
mitochondrial channelopathy, or pyruvate dehydrogenase complex
deficiency (PDCD/PDH);
[0313] diabetic neuropathy;
[0314] chemotherapy-induced peripheral neuropathy; and/or
[0315] crush injury, spinal cord injury, traumatic brain injury,
stroke, optic nerve injury, and related conditions that involve
axonal disconnection.
[0316] Clause 24. A composition comprising the compound of any one
of clauses 6 to 17 and a pharmaceutically acceptable excipient.
[0317] The present disclosure is also directed to the following
embodiments:
[0318] A. Methods for treating a mitochondria-associated disease,
disorder, or condition. The methods comprise: administering a
therapeutically effective amount of a composition comprising one or
more of mitofusin activator or a pharmaceutically acceptable salt
thereof to a subject having or suspected of having a
mitochondria-associated disease, disorder or condition, the
mitofusin activator having a structure represented by the following
formula:
##STR00032##
[0319] wherein: [0320] Z is cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl; [0321] R.sup.2 and R.sup.3 are independently selected
from H, F, alkyl, and C.sub.3-7 cycloalkyl, or R.sup.2 and R.sup.3
taken together form a C.sub.3-7 cycloalkyl or heterocycloalkyl;
[0322] R.sup.4 and R.sup.5 are independently selected from H, F,
alkyl, and C.sub.3-7 cycloalkyl, or R.sup.4 and R.sup.5 taken
together form a C.sub.3-7 cycloalkyl or heterocycloalkyl; [0323] Y
is O, CR.sup.6R.sup.7, CR.sup.8.dbd.CR.sup.9, C.ident.C,
cycloalkyl, heterocycloalkyl, aryl, heteroaryl, NR.sup.8, S,
SO.sub.2, SONR.sup.9, NR.sup.9SO.sub.2, NR.sup.8CO, CONR.sup.8, or
NR.sup.8CONR.sup.9; [0324] R.sup.6 is H, F, alkyl, or cycloalkyl,
R.sup.7 is H, F, alkyl, or cycloalkyl, or R.sup.6 and R.sup.7 taken
together form a C.sub.3-7 cycloalkyl or heterocycloalkyl; [0325]
R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl, R.sup.9 is H, alkyl,
or C.sub.3-7 cycloalkyl, or R.sup.8 and R.sup.9 taken together form
a C.sub.3-7 cycloalkyl; [0326] o is 0, 1, 2, 3, 4 or 5; [0327] p is
0 or 1; and [0328] q is 0, 1, 2, 3, 4 or 5, provided that if Y is
cycloalkyl and p is 1 the sum of o+p+q is not less than 3 or
greater than 5, and otherwise the sum of o+p+q is 5.
[0329] B. Compositions comprising a mitofusin activator or a
pharmaceutically acceptable salt thereof. The compositions
comprise: a mitofusin activator or a pharmaceutically acceptable
salt thereof, the mitofusin activator having a structure
represented by the following formula:
##STR00033##
[0330] wherein: [0331] Z is cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl; [0332] R.sup.2 and R.sup.3 are independently selected
from H, F, alkyl, and C.sub.3-7 cycloalkyl, or R.sup.2 and R.sup.3
taken together form a C.sub.3-7 cycloalkyl or heterocycloalkyl;
[0333] R.sup.4 and R.sup.5 are independently selected from H, F,
alkyl, and C.sub.3-7 cycloalkyl, or R.sup.4 and R.sup.5 taken
together form a C.sub.3-7 cycloalkyl or heterocycloalkyl; [0334] Y
is O, CR.sup.6R.sup.7, CR.sup.8.dbd.CR.sup.9, C.ident.C,
cycloalkyl, heterocycloalkyl, aryl, heteroaryl, NR.sup.8, S,
SO.sub.2, SONR.sup.9, NR.sup.9SO.sub.2, NR.sup.8CO, CONR.sup.8, or
NR.sup.8CONR.sup.9; [0335] R.sup.6 is H, F, alkyl, or cycloalkyl,
R.sup.7 is H, F, alkyl, or cycloalkyl, or R.sup.6 and R.sup.7 taken
together form a C.sub.3-7 cycloalkyl or heterocycloalkyl; [0336]
R.sup.8 is H, alkyl, or C.sub.3-7 cycloalkyl, R.sup.9 is H, alkyl,
or C.sub.3-7 cycloalkyl, or R.sup.8 and R.sup.9 taken together form
a C.sub.3-7 cycloalkyl; [0337] o is 0, 1, 2, 3, 4 or 5; [0338] p is
0 or 1; and [0339] q is 0, 1, 2, 3, 4 or 5, provided that if Y is
cycloalkyl and p is 1 the sum of o+p+q is not less than 3 or
greater than 5, and otherwise the sum of o+p+q is 5.
[0340] Embodiments A and B may include one or more of the following
elements:
[0341] Element 1: wherein:
[0342] Z is aryl or heteroaryl;
[0343] Y is O, CR.sup.6R.sup.7, or cycloalkyl;
[0344] R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, and R.sup.7 are
independently selected from H and alkyl; and
[0345] p is 1.
[0346] Element 2: wherein:
[0347] Y is cyclopropyl or cyclobutyl;
[0348] R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are H;
[0349] o is 0, 1, 2, or 3;
[0350] p is 1; and
[0351] q is 0, 1, 2 or 3.
[0352] Element 3: wherein:
[0353] Y is O or CH.sub.2;
[0354] R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are H; and
[0355] p is 1.
[0356] Element 4: wherein Z is phenyl or heteroaryl, the heteroaryl
containing 1 to 4 atoms independently selected from nitrogen,
oxygen and sulfur, and the phenyl or the heteroaryl having 0 to 4
substituents independently selected from R.sup.8, OR.sup.8, Cl, F,
CN, CF.sub.3, NR.sup.8R.sup.9, SO.sub.2NR.sup.8R.sup.9,
NR.sup.8SO.sub.2R.sup.9, SO.sub.2R.sup.9, CONR.sup.8R.sup.10,
NR.sup.8COR.sup.10, C.sub.3-7 cycloalkyl, and heterocycloalkyl;
wherein R.sup.10 is alkyl or C.sub.3-7 cycloalkyl.
[0357] Element 5: Z is phenyl, 2-pyridinyl, 3-pyridinyl,
4-pyridinyl, 6-pyrimidinyl, 5-pyrimidinyl, 4-pyrimidinyl or
2-pyrimidinyl, Z having 0 to 2 substituents independently selected
from R.sup.8, OR.sup.8, Cl, F, CN, CF.sub.3, NR.sup.8R.sup.9,
SO.sub.2R.sup.10, CONR.sup.8R.sup.9, and NR.sup.8COR.sup.10;
wherein R.sup.10 is alkyl or C.sub.3-7 cycloalkyl.
[0358] Element 6: wherein the mitofusin activator comprises a
trans-stereoisomer 6-phenylhexanamide derivative or a
pharmaceutically acceptable salt thereof; wherein the mitofusin
activator stimulates mitochondrial fusion, increases mitochondrial
fitness, and enhances mitochondrial subcellular transport.
[0359] Element 7: wherein the mitofusin activator has a structure
represented by the following formula:
##STR00034##
or a pharmaceutically acceptable salt, tautomer, or stereoisomer
thereof; wherein R.sup.1 is a non-, mono-, or poly-substituted
C.sub.3-8 cycloalkyl, C.sub.3-8 heteroaryl, C.sub.3-8 aryl, or
C.sub.3-8 heterocyclyl.
[0360] Element 8: wherein R.sup.1 is
##STR00035##
[0361] Element 9: wherein the mitofusin activator has a structure
represented by one or more of the following formulas:
##STR00036##
[0362] Element 10: wherein the mitochondria-associated disease,
disorder or condition is a peripheral nervous system (PNS) or
central nervous system (CNS) genetic or non-genetic disorder,
physical damage, and/or chemical injury.
[0363] Element 11: wherein the PNS or CNS disorder is selected from
any one or a combination of a chronic neurodegenerative condition
wherein mitochondrial fusion, fitness, or trafficking are impaired;
a disease or disorder associated with mitofusin 1 (MFN1) or
mitofusin 2 (MFN2) dysfunction; a disease associated with
mitochondrial fragmentation, dysfunction, or dysmotility; a
degenerative neuromuscular condition such as Charcot-Marie-Tooth
disease, Amyotrophic Lateral Sclerosis, Huntington's disease,
Alzheimer's disease, Parkinson's disease; hereditary motor and
sensory neuropathy, autism, autosomal dominant optic atrophy
(ADOA), muscular dystrophy, Lou Gehrig's disease, cancer,
mitochondrial myopathy, diabetes mellitus and deafness (DAD),
Leber's hereditary optic neuropathy (LHON), Leigh syndrome,
subacute sclerosing encephalopathy, neuropathy, ataxia, retinitis
pigmentosa, and ptosis (NARP), myoneurogenic gastrointestinal
encephalopathy (MNGIE), myoclonic epilepsy with ragged red fibers
(MERRF), mitochondrial myopathy, encephalomyopathy, lactic
acidosis, stroke-like symptoms (MELAS), mtDNA depletion,
mitochondrial neurogastrointestinal encephalomyopathy (MNGIE),
dysautonomic mitochondrial myopathy, mitochondrial channelopathy,
or pyruvate dehydrogenase complex deficiency (PDCD/PDH); diabetic
neuropathy; chemotherapy-induced peripheral neuropathy; crush
injury, spinal cord injury (SCI), traumatic brain injury, stroke,
optic nerve injury, and related conditions that involve axonal
disconnection; and any combination thereof.
[0364] Element 12: wherein the composition further comprises a
pharmaceutically acceptable excipient.
[0365] All documents described herein are incorporated by reference
herein for purposes of all jurisdictions where such practice is
allowed, including any priority documents and/or testing procedures
to the extent they are not inconsistent with this text. As is
apparent from the foregoing general description and the specific
embodiments, while forms of the disclosure have been illustrated
and described, various modifications can be made without departing
from the spirit and scope of the disclosure. Accordingly, it is not
intended that the disclosure be limited thereby. For example, the
compositions described herein may be free of any component, or
composition not expressly recited or disclosed herein. Any method
may lack any step not recited or disclosed herein. Likewise, the
term "comprising" is considered synonymous with the term
"including." Whenever a method, composition, element or group of
elements is preceded with the transitional phrase "comprising," it
is understood that we also contemplate the same composition or
group of elements with transitional phrases "consisting essentially
of," "consisting of," "selected from the group of consisting of,"
or "is" preceding the recitation of the composition, element, or
elements and vice versa. The term "and/or" as used in a phrase such
as "A and/or B" herein is intended to include "A and B," "A or B,"
"A," and "B." Numerical ranges used herein include the numbers
recited in the range. For example, the numerical range "from 1 wt %
to 10 wt %" includes 1 wt % and 10 wt % within the recited
range.
[0366] Unless otherwise indicated, all numbers expressing
quantities of ingredients, properties such as molecular weight,
reaction conditions, and so forth used in the present specification
and associated claims are to be understood as being modified in all
instances by the term "about." Accordingly, unless indicated to the
contrary, the numerical parameters set forth in the following
specification and attached 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 claim, each numerical parameter
should at least be construed in light of the number of reported
significant digits and by applying ordinary rounding
techniques.
[0367] Whenever a numerical range with a lower limit and an upper
limit is disclosed, any number and any included range falling
within the range is specifically disclosed. In particular, every
range of values (of the form, "from about a to about b," or,
equivalently, "from approximately a to b," or, equivalently, "from
approximately a-b") disclosed herein is to be understood to set
forth every number and range encompassed within the broader range
of values. Also, the terms in the claims have their plain, ordinary
meaning unless otherwise explicitly and clearly defined by the
patentee. Moreover, the indefinite articles "a" or "an," as used in
the claims, are defined herein to mean one or more than one of the
element that it introduces.
[0368] One or more illustrative embodiments are presented herein.
Not all features of a physical implementation are described or
shown in this application for the sake of clarity. It is understood
that in the development of a physical embodiment of the present
disclosure, numerous implementation-specific decisions must be made
to achieve the developer's goals, such as compliance with
system-related, business-related, government-related and other
constraints, which vary by implementation and from time to time.
While a developer's efforts might be time-consuming, such efforts
would be, nevertheless, a routine undertaking for one of ordinary
skill in the art and having benefit of this disclosure.
[0369] Therefore, the present disclosure is well adapted to attain
the ends and advantages mentioned as well as those that are
inherent therein. The particular embodiments disclosed above are
illustrative only, as the present disclosure may be modified and
practiced in different but equivalent manners apparent to one
having ordinary skill in the art and having the benefit of the
teachings herein. Furthermore, no limitations are intended to the
details of construction or design herein shown, other than as
described in the claims below. It is therefore evident that the
particular illustrative embodiments disclosed above may be altered,
combined, or modified and all such variations are considered within
the scope and spirit of the present disclosure. The embodiments
illustratively disclosed herein suitably may be practiced in the
absence of any element that is not specifically disclosed herein
and/or any optional element disclosed herein.
* * * * *