U.S. patent application number 13/967283 was filed with the patent office on 2014-02-20 for morpholinoalkyl fumarate compounds, pharmaceutical compositions, and methods of use.
This patent application is currently assigned to XenoPort, Inc.. The applicant listed for this patent is XenoPort, Inc.. Invention is credited to Kenneth C. Cundy, Suresh K. Manthati, David J. Wustrow.
Application Number | 20140051705 13/967283 |
Document ID | / |
Family ID | 47720772 |
Filed Date | 2014-02-20 |
United States Patent
Application |
20140051705 |
Kind Code |
A1 |
Cundy; Kenneth C. ; et
al. |
February 20, 2014 |
Morpholinoalkyl Fumarate Compounds, Pharmaceutical Compositions,
and Methods of Use
Abstract
Morpholinoalkyl fumarates, pharmaceutical compositions
comprising the morpholinoalkyl fumarates, and methods of using
morpholinoalkyl fumarates and pharmaceutical compositions for
treating neurodegenerative, inflammatory, and autoimmune disorders
including multiple sclerosis, psoriasis, irritable bowel disorder,
ulcerative colitis, arthritis, chronic obstructive pulmonary
disease, asthma, Parkinson's disease, Huntington's disease, and
amyotrophic lateral sclerosis are disclosed.
Inventors: |
Cundy; Kenneth C.; (Redwood
City, CA) ; Manthati; Suresh K.; (Sunnyvale, CA)
; Wustrow; David J.; (Los Gatos, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
XenoPort, Inc. |
Santa Clara |
CA |
US |
|
|
Assignee: |
XenoPort, Inc.
Santa Clara
CA
|
Family ID: |
47720772 |
Appl. No.: |
13/967283 |
Filed: |
August 14, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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13761864 |
Feb 7, 2013 |
|
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13967283 |
|
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61595835 |
Feb 7, 2012 |
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Current U.S.
Class: |
514/239.2 ;
544/171 |
Current CPC
Class: |
C07D 265/30 20130101;
A61P 25/16 20180101; A61P 29/00 20180101; A61P 11/06 20180101; C07D
295/088 20130101; A61P 25/00 20180101; A61P 25/14 20180101; A61P
17/06 20180101; A61P 37/06 20180101; A61P 11/00 20180101; A61P
37/00 20180101; A61P 1/04 20180101; A61P 19/02 20180101 |
Class at
Publication: |
514/239.2 ;
544/171 |
International
Class: |
C07D 295/088 20060101
C07D295/088 |
Claims
1-31. (canceled)
32. A compound according to the formula: ##STR00127## or a
pharmaceutically acceptable salt thereof; wherein: n is an integer
from 2 to 6; and Q is a tertiary amine moiety.
33. A compound of claim 32, wherein n is 2.
34. A compound of claim 32, wherein Q is a morpholino group.
35. A compound of claim 32, wherein the compound is a
pharmaceutically acceptable salt.
36. A compound according to claim 32, wherein the compound is a HCl
salt.
37. A pharmaceutical composition comprising a pharmaceutically
acceptable vehicle and a therapeutically effective amount of a
compound of claim 32.
38. A method for treating in a mammal in need thereof a disease or
condition which comprises administering to the mammal an effective
disease-treating or condition-treating amount of a compound of
claim 32.
39. The method of claim 38, wherein the disease or condition is
selected from a neurodegenerative disease, an inflammatory disease,
and an autoimmune disease.
40. The method of claim 38, wherein the disease or condition is
selected from multiple sclerosis, psoriasis, irritable bowel
disorder, ulcerative colitis, arthritis, chronic obstructive
pulmonary disease, asthma, Parkinson's disease, Huntington's
disease, and amyotrophic lateral sclerosis.
41. The method of claim 38, wherein the disease or condition is
multiple sclerosis.
42. The method of claim 38, wherein the disease or condition is
psoriasis.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 13/761,864, filed Feb. 7, 2013, which claims
the benefit of U.S. Provisional Patent Application No. 61/595,835
filed Feb. 7, 2012, both of which are incorporated herein by
reference in their entirety.
FIELD
[0002] Disclosed herein are morpholinoalkyl fumarates,
pharmaceutical compositions comprising the morpholinoalkyl
fumarates, and methods of using said morpholinoalkyl fumarates and
pharmaceutical compositions thereof for treating neurodegenerative,
inflammatory, and autoimmune diseases including multiple sclerosis,
psoriasis, irritable bowel disorder, ulcerative colitis, arthritis,
chronic obstructive pulmonary disease, asthma, Parkinson's disease,
Huntington's disease, and amyotrophic lateral sclerosis.
BACKGROUND
[0003] Fumaric acid esters (FAEs) are approved in Germany for the
treatment of psoriasis, are being evaluated in the United States
for the treatment of psoriasis and multiple sclerosis, and have
been proposed for use in treating a wide range of immunological,
autoimmune, and inflammatory diseases and conditions.
[0004] FAEs and other fumaric acid derivatives have been proposed
for use in treating a wide-variety of diseases and conditions
involving immunological, autoimmune, and/or inflammatory processes
including psoriasis (Joshi and Strebel, WO 1999/49858; U.S. Pat.
No. 6,277,882; Mrowietz and Asadullah, Trends Mol Med 2005, 111(1),
43-48; and Yazdi and Mrowietz, Clinics Dermatology 2008, 26,
522-526); asthma and chronic obstructive pulmonary diseases (Joshi
et al., WO 2005/023241 and US 2007/0027076); cardiac insufficiency
including left ventricular insufficiency, myocardial infarction and
angina pectoris (Joshi et al., WO 2005/023241; Joshi et al., US
2007/0027076); mitochondrial and neurodegenerative diseases such as
Parkinson's disease, Alzheimer's disease, Huntington's disease,
retinopathia pigmentosa and mitochondrial encephalomyopathy (Joshi
and Strebel, WO 2002/055063, US 2006/0205659, U.S. Pat. No.
6,509,376, U.S. Pat. No. 6,858,750, and U.S. Pat. No. 7,157,423);
transplantation (Joshi and Strebel, WO 2002/055063, US
2006/0205659, U.S. Pat. No. 6,359,003, U.S. Pat. No. 6,509,376, and
U.S. Pat. No. 7,157,423; and Lehmann et al., Arch Dermatol Res
2002, 294, 399-404); autoimmune diseases (Joshi and Strebel, WO
2002/055063, U.S. Pat. No. 6,509,376, U.S. Pat. No. 7,157,423, and
US 2006/0205659) including multiple sclerosis (MS) (Joshi and
Strebel, WO 1998/52549 and U.S. Pat. No. 6,436,992; Went and
Lieberburg, US 2008/0089896; Schimrigk et al., Eur J Neurology
2006, 13, 604-610; and Schilling et al., Clin Experimental
Immunology 2006, 145, 101-107); ischemia and reperfusion injury
(Joshi et al., US 2007/0027076); advanced glycation end products
(AGE)-induced genome damage (Heidland, WO 2005/027899);
inflammatory bowel diseases such as Crohn's disease and ulcerative
colitis; arthritis; and others (Nilsson et al., WO 2006/037342 and
Nilsson and Muller, WO 2007/042034).
[0005] The mechanism of action of fumaric acid esters is believed
to be mediated by pathways associated with the immunological
response. For example, FAEs invoke a shift from a Th1 to Th2 immune
response, favorably altering the cytokine profile; inhibit
cytokine-induced expression of adhesion molecules such as VCAM-1,
ICAM-1 and E-selectin, thereby reducing immune cell extravasation;
and deplete lymphocytes through apoptotic mechanisms (Lehmann et
al., J Investigative Dermatology 2007, 127, 835-845; Gesser et al.,
J Investigative Dermatology 2007, 127, 2129-2137; Vandermeeren et
al., Biochm Biophys Res Commun 1997, 234, 19-23; and Treumer et
al., J Invest Dermatol 2003, 121, 1383-1388).
[0006] Recent studies suggest that FAEs are inhibitors of
NF-.kappa.B activation, a transcription factor that regulates the
inducible expression of proinflammatory mediators (D'Acquisto et
al., Molecular Interventions 2002, 2(1), 22-35). Accordingly, FAEs
have been proposed for use in treating NF-.kappa.B mediated
diseases (Joshi et al., WO 2002/055066; and Joshi and Strebel, WO
2002/055063, US 2006/0205659, U.S. Pat. No. 7,157,423 and U.S. Pat.
No. 6,509,376). Inhibitors of NF-.kappa.B activation have also been
shown to be useful in angiostatic therapy (Tabruyn and Griffioen,
Angiogenesis 2008, 11, 101-106), inflammatory bowel disease (Atreya
et al., J Intern Med 2008, 263(6), 591-6); and in animal models of
diseases involving inflammation including neutrophilic alveolitis,
asthma, hepatitis, inflammatory bowel disease, neurodegeneration,
ischemia/reperfusion, septic shock, glomerulonephritis, and
rheumatoid arthritis (D'Acquisto et al., Molecular Interventions
2002, 2(1), 22-35).
[0007] Studies also suggest that NF-.kappa.B inhibition by FAEs may
be mediated by interaction with tumor necrosis factor (TNF)
signaling. Dimethyl fumarate inhibits TNF-induced tissue factor
mRNA and protein expression and TNF-induced DNA binding of
NF-.kappa.B proteins, and inhibits the TNF-induced nuclear entry of
activated NF-.kappa.B proteins thereby inhibiting inflammatory gene
activation (Loewe et al., J Immunology 2002, 168, 4781-4787). TNF
signaling pathways are implicated in the pathogenesis of
immune-mediated inflammatory diseases such as rheumatoid arthritis,
Crohn's disease, psoriasis, psoriatic arthritis, juvenile
idiopathic arthritis, and ankylosing spondylitis (Tracey et al.,
Pharmacology & Therapetuics 2008, 117, 244-279).
[0008] FUMADERM.RTM., an enteric coated tablet containing a salt
mixture of ethyl hydrogen fumarate and dimethyl fumarate (DMF) (2),
which is rapidly hydrolyzed to methyl hydrogen fumarate (MHF) (1)
in vivo and is regarded as the main bioactive metabolite, was
approved in Germany in 1994 for the treatment of psoriasis.
##STR00001##
[0009] FUMADERM.RTM. is dosed three times/day with 1-2 grams/day
administered for the treatment of psoriasis. FUMADERM.RTM. exhibits
a high degree of interpatient variability with respect to drug
absorption and food strongly reduces bioavailability. Absorption is
thought to occur in the small intestine with peak levels achieved
5-6 hours after oral administration. Significant side effects occur
in 70-90% of patients (Brewer and Rogers, Clin Expt'l Dermatology
2007, 32, 246-49; and Hoefnagel et al., Br J Dermatology 2003, 149,
363-369). Side effects of current FAE therapy include
gastrointestinal upset including nausea, vomiting, diarrhea, and
transient flushing of the skin. Also, DMF exhibits poor aqueous
solubility.
[0010] Fumaric acid derivatives (Joshi and Strebel, WO 2002/055063,
US 2006/0205659, and U.S. Pat. No. 7,157,423 (amide compounds and
protein-fumarate conjugates); Joshi et al., WO 2002/055066 and
Joshi and Strebel, U.S. Pat. No. 6,355,676 (mono and dialkyl
esters); Joshi and Strebel, WO 2003/087174 (carbocyclic and
oxacarbocylic compounds); Joshi et al., WO 2006/122652
(thiosuccinates); Joshi et al., US 2008/0233185 (dialkyl and diaryl
esters)) and salts (Nilsson et al., US 2008/0004344) have been
developed in an effort to overcome the deficiencies of current FAE
therapy. Controlled release pharmaceutical compositions comprising
fumaric acid esters are disclosed by Nilsson and Muller, WO
2007/042034. Glycolamide ester prodrugs are described by Nielsen
and Bundgaard, J Pharm Sci 1988, 77(4), 285-298.
[0011] Flachsmann et al., U.S. Pat. No. 7,638,118, discloses
compounds having the following chemical formula:
##STR00002##
wherein: Z is --OR.sup.2 or --Y--(R--NR.sup.3R.sup.4).sub.n; R can
be a linear or branched C.sub.2-9 alkyl; R.sup.2 can be a linear or
branched C.sub.1-8 alkyl; R.sup.3 and R.sup.4, together with the
nitrogen atom to which they are bonded, can form an aromatic
heterocyclic ring such as a morpholinyl ring; and when n is 1, Y
can be oxygen.
[0012] The compounds are disclosed to be useful for neutralizing
odors.
[0013] Morpholinoalkyl ester prodrugs of the non-steroidal
anti-inflammatory drug niflumic acid exhibit unexpectedly high
protection from gastric irritation and ulcerogenicity compared to
the parent acid drug (Talath and Gadad, Arzneimittelforschung 2006,
56(11), 744-52). The protective effect is believed to involve
absorption of the intact prodrug, which reduces local gastric
exposure. Although glycolamide esters of niflumic acid have been
synthesized in an effort to improve the biocompatibility of
niflumic aid, the effects on gastrointestinal irritation in humans
does not appear to have been reported (Talath et al,
Arzneimittelforschung 2006, 56(9), 631-9; Gadad et al.,
Arzneimittelforschung 2002, 52(11), 817-21; Benoit et al., Rev.
Odontostomatol Midi Fr. 1975, 4, 249-61; and Los et al., Farmaco
Sci. 1981 36(5), 372-85). However, the morpholinoalkyl esters, and
specifically the morpholinopropyl and morpholinobutyl esters of
niflumic acid were identified as exhibiting the best combination of
stability, in vivo anti-inflammatory activity, and low
ulcerogenicity in rats (Talath and Gadad, Arzneimittelforschung
2006, 56(11), 744-52).
[0014] Gangakhedkar et al., U.S. Patent Publication No.
2010/0048651, discloses compounds having the following chemical
formula:
##STR00003##
wherein: R.sup.1 and R.sup.2 are independently chosen from
hydrogen, C.sub.1-6 alkyl, and substituted C.sub.1-6 alkyl; R.sup.3
and R.sup.4, together with the nitrogen to which they are bonded,
can form a C.sub.5-10 heteroaryl ring such as a morpholino ring;
and R.sup.5 can be hydrogen, methyl, ethyl, and C.sub.3-6 alkyl;
and pharmaceutical compositions containing such compounds for the
treatment of diseases including psoriasis, multiple sclerosis, an
inflammatory bowel disease, asthma, chronic obstructive pulmonary
disease, Parkinson's disease, Huntington's disease, amyotrophic
lateral sclerosis (ALS), and arthritis. Compounds in which
--NR.sup.3R.sup.4 is a morpholino ring are disclosed in Example 3
(methyl 2-morpholin-4-yl-2-oxoethyl (2E)but-2-ene-1,4-dioate),
Example 28 (methyl 1-methyl-2-morpholin-4-yl-2-oxoethyl
(2E)but-2-ene-1,4-dioate), Example 31
((1S)-1-methyl-2-morpholin-4-yl-2-oxoethyl methyl
(2E)but-2-ene-1,4-dioate), and Example 47
((2E)-3-[(2-morpholin-4-yl-2-oxoethyl)oxycarbonyl]prop-2-enoic
acid).
SUMMARY
[0015] Morpholinoalkyl fumarates having high gastrointestinal
permeability and/or absorption, improved solubility, ordered
hydrolysis (i.e., preferential cleavage of promoieties), and
minimal cleavage in the gut lumen or enterocyte cytoplasm are
desirable. Such morpholinoalkyl fumarates, which provide higher
oral bioavailability and plasma levels of the parent compound, an
alkyl hydrogen fumarate, e.g., MHF, and/or other metabolites of the
morpholinoalkyl fumarates, may: enhance the efficacy/responder rate
compared to present fumaric acid esters; facilitate the use of
lower doses, reduce dosing frequency, and standardize dosing
regimens; reduce food effects; reduce gastrointestinal side
effects/toxicity; and reduce interpatient treatment
variability.
[0016] Morpholinoalkyl esters of alkyl hydrogen fumarates and
hydrogen fumarates having reduced gastrointestinal side effects are
disclosed.
[0017] In a first aspect, pharmaceutical compositions comprising a
pharmaceutically acceptable vehicle and a therapeutically effective
amount of a compound of Formula (I):
##STR00004##
or a pharmaceutically acceptable salt thereof, are provided,
wherein:
[0018] n is an integer from 2 to 6;
[0019] R.sup.1 is chosen from methyl, ethyl, C.sub.3-6 alkyl,
and
##STR00005##
and
[0020] m is an integer from 2 to 6.
[0021] Such pharmaceutical compositions are useful for treating
neurodegenerative, inflammatory and autoimmune diseases and
disorders including, for example, multiple sclerosis, psoriasis,
irritable bowel disorder, ulcerative colitis, arthritis, chronic
obstructive pulmonary disease, asthma, Parkinson's disease,
Huntington's disease, and amyotrophic lateral sclerosis.
[0022] In a second aspect, methods of treating a disease in a
patient are provided comprising administering to a patient in need
of such treatment a pharmaceutical composition comprising a
therapeutically effective amount of a compound of Formula (I). In
certain embodiments, the disease is chosen from a neurodegenerative
disease, an inflammatory disease, and an autoimmune disease
including, for example, multiple sclerosis, psoriasis, irritable
bowel disorder, ulcerative colitis, arthritis, chronic obstructive
pulmonary disease, asthma, Parkinson's disease, Huntington's
disease, and amyotrophic lateral sclerosis.
[0023] In a third aspect, pharmaceutical compositions comprising a
pharmaceutically acceptable vehicle and a therapeutically effective
amount of a compound of Formula (II):
##STR00006##
or a pharmaceutically acceptable salt thereof, are provided,
wherein n is an integer from 2 to 6.
[0024] In a fourth aspect, methods of treating a disease in a
patient are provided comprising administering to a patient in need
of such treatment a pharmaceutical composition comprising a
therapeutically effective amount of a compound of Formula (II). In
certain embodiments, the disease is chosen from a neurodegenerative
disease, an inflammatory disease, and an autoimmune disease
including, for example, multiple sclerosis, psoriasis, irritable
bowel disorder, ulcerative colitis, arthritis, chronic obstructive
pulmonary disease, asthma, Parkinson's disease, Huntington's
disease, and amyotrophic lateral sclerosis.
[0025] In a specific aspect, provided here are compounds according
to Formula (I):
##STR00007##
[0026] or a pharmaceutically acceptable salt thereof;
wherein:
[0027] n is an integer from 2 to 6;
[0028] R.sup.1 is selected from H, methyl, ethyl, C.sub.3-6 alkyl,
and
##STR00008##
and
[0029] m is an integer from 2 to 6;
[0030] provided that
[0031] i) when n is 2, and R.sup.1 is
##STR00009##
[0032] then m is 3, 4, 5, or 6; and
[0033] ii) when R.sup.1 is H, then n is 4, 5, or 6.
[0034] Such compounds are useful for treating neurodegenerative,
inflammatory and autoimmune diseases and disorders including, for
example, multiple sclerosis, psoriasis, irritable bowel disorder,
ulcerative colitis, arthritis, chronic obstructive pulmonary
disease, asthma, Parkinson's disease, Huntington's disease, and
amyotrophic lateral sclerosis.
[0035] In a particular embodiment, with respect to the
pharmaceutical compositions and compounds according to Formula (I),
the compound is according to Formula (IIIc):
##STR00010##
or a pharmaceutically acceptable salt thereof.
[0036] In a more particular embodiment, with respect to the
pharmaceutical compositions and compounds according to Formula (I),
the compound is a HCl salt of a compound according to Formula
(IIIc).
DETAILED DESCRIPTION
Figures
[0037] FIG. 1: GI Effect of the Compounds of the Disclosure--FIG. 1
depicts the GI irritation score of exemplary compounds along with
comparative compounds (CC-1 and DMF) at an oral dose of 180 mg
eq/kg, dosed per day for 4 days in rats.
DEFINITIONS
[0038] A dash ("--") that is not between two letters or symbols is
used to indicate a point of attachment for a moiety or substituent.
For example, --CONH.sub.2 is bonded through the carbon atom.
[0039] "Alkyl" refers to a saturated or unsaturated, branched, or
straight-chain, monovalent hydrocarbon radical derived by the
removal of one hydrogen atom from a single carbon atom of a parent
alkane, alkene, or alkyne. Examples of alkyl groups include, for
example, methyl; ethyls such as ethanyl, ethenyl, and ethynyl;
propyls such as propan-1-yl, propan-2-yl, prop-1-en-1-yl,
prop-1-en-2-yl, prop-2-en-1-yl (allyl), prop-1-yn-1-yl,
prop-2-yn-1-yl, etc.; butyls such as butan-1-yl, butan-2-yl,
2-methyl-propan-1-yl, 2-methyl-propan-2-yl, but-1-en-1-yl,
but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl,
but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl,
but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the
like.
[0040] The term "alkyl" includes groups having any degree or level
of saturation, i.e., groups having exclusively single carbon-carbon
bonds, groups having one or more double carbon-carbon bonds, groups
having one or more triple carbon-carbon bonds, and groups having
combinations of single, double, and triple carbon-carbon bonds.
Where a specific level of saturation is intended, the terms
alkanyl, alkenyl, or alkynyl are used. In certain embodiments, an
alkyl group can have from 1 to 10 carbon atoms (C.sub.1-10), in
certain embodiments, from 1 to 6 carbon atoms (C.sub.1-6), in
certain embodiments from 1 to 4 carbon atoms (C.sub.1-4), in
certain embodiments, from 1 to 3 carbon atoms (C.sub.1-3), and in
certain embodiments, from 1 to 2 carbon atoms (C.sub.1-2). In
certain embodiments, alkyl is methyl, in certain embodiments,
ethyl, and in certain embodiments, n-propyl or isopropyl.
[0041] "Compounds" of Formula (I) and Formula (II) disclosed herein
include any specific compounds within this formula. Compounds may
be identified either by their chemical structure and/or chemical
name. Compounds are named using Chemistry 4-D Draw Pro, version
7.01c (ChemInnovation Software, Inc., San Diego, Calif.). When the
chemical structure and chemical name conflict, the chemical
structure is determinative of the identity of the compound. The
compounds described herein may comprise one or more chiral centers
and/or double bonds and therefore may exist as stereoisomers such
as double-bond isomers (i.e., geometric isomers), enantiomers, or
diastereomers. Accordingly, any chemical structures within the
scope of the specification depicted, in whole or in part, with a
relative configuration encompass all possible enantiomers and
stereoisomers of the illustrated compounds including the
stereoisomeric ally pure form (e.g., geometrically pure,
enantiomerically pure, or diastereomerically pure) and enantiomeric
and stereoisomeric mixtures. Enantiomeric and stereoisomeric
mixtures may be resolved into their component enantiomers or
stereoisomers using separation techniques or chiral synthesis
techniques well known to those skilled in the art. Compounds of
Formula (I) and Formula (II) include, for example, optical isomers
of compounds of Formula (I) and Formula (II), racemates thereof,
and other mixtures thereof. In such embodiments, a single
enantiomer or diastereomer, i.e., optically active form can be
obtained by asymmetric synthesis or by resolution of the racemates.
Resolution of the racemates may be accomplished, for example, by
methods such as crystallization in the presence of a resolving
agent, or chromatography using, for example, chiral stationary
phases. Notwithstanding the foregoing, in compounds of Formula (I)
and Formula (II) the configuration of the illustrated double bond
is only in the E configuration (i.e., trans configuration).
[0042] Compounds of Formula (I) and Formula (II) also include
isotopically labeled compounds where one or more atoms have an
atomic mass different from the atomic mass conventionally found in
nature. Examples of isotopes that may be incorporated into the
compounds disclosed herein include, for example, .sup.2H, .sup.3H,
.sup.11C, .sup.13C, .sup.14C, .sup.15N .sup.18O, .sup.17O, etc.
[0043] Compounds may exist in unsolvated forms as well as solvated
forms, including hydrated forms and as N-oxides. In general,
compounds disclosed herein may be free acid, hydrated, solvated, or
N-oxides. Certain compounds may exist in multiple crystalline,
co-crystalline, or amorphous forms. Compounds of Formula (I) and
Formula (II) include pharmaceutically acceptable salts thereof or
pharmaceutically acceptable solvates of the free acid form of any
of the foregoing, as well as crystalline forms of any of the
foregoing.
[0044] Compounds of Formula (I) and Formula (II) also include
solvates. A solvate refers to a molecular complex of a compound
with one or more solvent molecules in a stoichiometric or
non-stoichiometric amount. Such solvent molecules include those
commonly used in the pharmaceutical art, which are known to be
innocuous to a patient, e.g., water, ethanol, and the like. A
molecular complex of a compound or moiety of a compound and a
solvent can be stabilized by non-covalent intra-molecular forces
such as, for example, electrostatic forces, van der Waals forces,
or hydrogen bonds. The term "hydrate" refers to a solvate in which
the one or more solvent molecules are water.
[0045] Further, when partial structures of the compounds are
illustrated, an asterisk (*) indicates the point of attachment of
the partial structure to the rest of the molecule.
[0046] "Disease" refers to a disease, disorder, condition, or
symptom of any of the foregoing.
[0047] "Drug" as defined under 21 U.S.C. .sctn.321(g)(1) means "(A)
articles recognized in the official United States Pharmacopoeia,
official Homeopathic Pharmacopoeia of the United States, or
official National Formulary, or any supplement to any of them; and
(B) articles intended for use in the diagnosis, cure, mitigation,
treatment, or prevention of disease in man or other animals; and
(C) articles (other than food) intended to affect the structure or
any function of the body of man or other animals . . . ."
[0048] "Leaving group" has the meaning conventionally associated
with it in synthetic organic chemistry, i.e., an atom or a group
capable of being displaced by a nucleophile and includes halogen
such as chloro, bromo, fluoro, and iodo; acyloxy, such as acetoxy
and benzoyloxy, alkoxycarbonylaryloxycarbonyl, mesyloxy, tosyloxy,
and trifluoromethanesulfonyloxy; aryloxy such as
2,4-dinitrophenoxy, methoxy, N,O-dimethylhydroxylamino,
p-nitrophenolate, imidazolyl, and the like.
[0049] "MHF" refers to methyl hydrogen fumarate, a compound having
the following chemical structure:
##STR00011##
This compound is also sometimes referred to as monomethyl
fumarate.
[0050] "Parent heteroaromatic ring system" refers to an aromatic
ring system in which one or more carbon atoms (and any associated
hydrogen atoms) are independently replaced with the same or
different heteroatom in such a way as to maintain the continuous
.pi.-electron system characteristic of aromatic systems and a
number of out-of-plane .pi.-electrons corresponding to the Huckel
rule (4n+2). Examples of heteroatoms to replace the carbon atoms
include, for example, N, P, O, S, and Si, etc. Specifically
included within the definition of "parent heteroaromatic ring
systems" are fused ring systems in which one or more of the rings
are aromatic and one or more of the rings are saturated or
unsaturated, such as, for example, arsindole, benzodioxan,
benzofuran, chromane, chromene, indole, indoline, xanthene, etc.
Examples of parent heteroaromatic ring systems include, for
example, arsindole, carbazole, .beta.-carboline, chromane,
chromene, cinnoline, furan, imidazole, indazole, indole, indoline,
indolizine, isobenzofuran, isochromene, isoindole, isoindoline,
isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole,
oxazole, perimidine, phenanthridine, phenanthroline, phenazine,
phthalazine, pteridine, purine, pyran, pyrazine, pyrazole,
pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine,
quinazoline, quinoline, quinolizine, quinoxaline, tetrazole,
thiadiazole, thiazole, thiophene, triazole, xanthene, thiazolidine,
oxazolidine, and the like.
[0051] "Patient" refers to a mammal, for example, a human.
[0052] "Pharmaceutically acceptable" refers to approved or
approvable by a regulatory agency of the Federal or a state
government or listed in the U.S. Pharmacopoeia or other generally
recognized pharmacopoeia for use in animals, and more particularly
in humans.
[0053] "Pharmaceutically acceptable salt" refers to a salt of a
compound that possesses the desired pharmacological activity of the
parent compound. Such salts include acid addition salts, formed
with inorganic acids such as hydrochloric acid, hydrobromic acid,
sulfuric acid, nitric acid, phosphoric acid, and the like; or
formed with organic acids such as acetic acid, propionic acid,
hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic
acid, lactic acid, malonic acid, succinic acid, malic acid, maleic
acid, fumaric acid, tartaric acid, citric acid, benzoic acid,
3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,
methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic
acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,
4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,
4-toluenesulfonic acid, camphorsulfonic acid,
4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic
acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary
butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic
acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic
acid, and the like; and salts formed when an acidic proton present
in the parent compound is replaced by a metal ion, e.g., an alkali
metal ion, an alkaline earth ion, or an aluminum ion; or
coordinates with an organic base such as ethanolamine,
diethanolamine, triethanolamine, N-methylglucamine, and the like.
In certain embodiments, a pharmaceutically acceptable salt is the
hydrochloride salt. In certain embodiments, a pharmaceutically
acceptable salt is the sodium salt.
[0054] "Pharmaceutically acceptable vehicle" refers to a
pharmaceutically acceptable diluent, a pharmaceutically acceptable
adjuvant, a pharmaceutically acceptable excipient, a
pharmaceutically acceptable carrier, or a combination of any of the
foregoing with which a compound provided by the present disclosure
may be administered to a patient, which does not destroy the
pharmacological activity thereof and which is non-toxic when
administered in doses sufficient to provide a therapeutically
effective amount of the compound or a pharmacologically active
metabolite thereof.
[0055] "Pharmaceutical composition" refers to a compound of Formula
(I), or a pharmaceutically acceptable salt thereof, or a compound
of Formula (II), or a pharmaceutically acceptable salt thereof, and
at least one pharmaceutically acceptable vehicle, with which the
compound of Formula (I), or a pharmaceutically acceptable salt
thereof, or the compound of Formula (II), or a pharmaceutically
acceptable salt thereof, is administered to a patient.
[0056] "Treating" or "treatment" of any disease refers to
reversing, alleviating, arresting, or ameliorating a disease or at
least one of the clinical symptoms of a disease, reducing the risk
of acquiring a disease or at least one of the clinical symptoms of
a disease, inhibiting the progress of a disease or at least one of
the clinical symptoms of the disease or reducing the risk of
developing a disease or at least one of the clinical symptoms of a
disease. "Treating" or "treatment" also refers to inhibiting a
disease, either physically, (e.g., stabilization of a discernible
symptom), physiologically, (e.g., stabilization of a physical
parameter), or both, and to inhibiting at least one physical
parameter that may or may not be discernible to the patient. In
certain embodiments, "treating" or "treatment" refers to delaying
the onset of a disease or at least one or more symptoms thereof in
a patient who may be exposed to or predisposed to a disease even
though that patient does not yet experience or display symptoms of
the disease.
[0057] "Therapeutically effective amount" refers to the amount of a
compound that, when administered to a subject for treating a
disease, or at least one of the clinical symptoms of a disease, is
sufficient to effect such treatment of the disease or symptom
thereof. The "therapeutically effective amount" may vary depending,
for example, on the compound, the disease and/or symptoms of the
disease, severity of the disease and/or symptoms of the disease,
the age, weight, and/or health of the patient to be treated, and
the judgment of the prescribing physician. An appropriate amount in
any given compound may be ascertained by those skilled in the art
and/or is capable of determination by routine experimentation.
[0058] "Therapeutically effective dose" refers to a dose that
provides effective treatment of a disease in a patient. A
therapeutically effective dose may vary from compound to compound
and/or from patient to patient, and may depend upon factors such as
the condition of the patient and the route of delivery. A
therapeutically effective dose may be determined in accordance with
routine pharmacological procedures known to those skilled in the
art.
[0059] Reference is now made in detail to certain embodiments of
compounds, compositions, and methods. The disclosed embodiments are
not intended to be limiting of the claims. To the contrary, the
claims are intended to cover all alternatives, modifications, and
equivalents.
Compounds
[0060] Certain embodiments provide a compound of Formula (I):
##STR00012##
or a pharmaceutically acceptable salt thereof, wherein:
[0061] n is an integer from 2 to 6;
R.sup.1 is chosen from methyl, ethyl, C.sub.3-6 alkyl, and
##STR00013##
and
[0062] m is an integer from 2 to 6.
[0063] Certain embodiments provide a compound of Formula (I):
##STR00014##
or a pharmaceutically acceptable salt thereof, wherein:
[0064] n is an integer from 2 to 6; and
[0065] R.sup.1 is chosen from methyl, ethyl, and C.sub.3-6
alkyl.
[0066] Certain embodiments provide a pharmaceutical composition
comprising a pharmaceutically acceptable vehicle and a
therapeutically effective amount of a compound of Formula (I):
##STR00015##
or a pharmaceutically acceptable salt thereof, wherein:
[0067] n is an integer from 2 to 6;
R.sup.1 is chosen from methyl, ethyl, C.sub.3-6 alkyl, and
##STR00016##
and m is an integer from 2 to 6.
[0068] Certain embodiments provide a pharmaceutical composition
comprising a pharmaceutically acceptable vehicle and a
therapeutically effective amount of a compound of Formula (I):
##STR00017##
or a pharmaceutically acceptable salt thereof, wherein:
[0069] n is an integer from 2 to 6; and
[0070] R.sup.1 is chosen from methyl, ethyl, and C.sub.3-6
alkyl.
[0071] In certain embodiments of a compound of Formula (I), n is 2,
n is 3, n is 4, n is 5, and in certain embodiments, n is 6.
[0072] In certain embodiments of a compound of Formula (I), m is 2,
m is 3, m is 4, m is 5, and in certain embodiments, m is 6.
[0073] In certain embodiments of a compound of Formula (I), n is 2
and m is 2; n is 2 and m is 3; n is 2 and m is 4; n is 2 and m is
5; and in certain embodiments, n is 2 and m is 6.
[0074] In certain embodiments of a compound of Formula (I), n is 3
and m is 2; n is 3 and m is 3; n is 3 and m is 4; n is 3 and m is
5; and in certain embodiments, n is 3 and m is 6.
[0075] In certain embodiments of a compound of Formula (I), n is 4
and m is 2; n is 4 and m is 3; n is 4 and m is 4; n is 4 and m is
5; and in certain embodiments, n is 4 and m is 6.
[0076] In certain embodiments of a compound of Formula (I), n is 5
and m is 2; n is 5 and m is 3; n is 5 and m is 4; n is 5 and m is
5; and in certain embodiments, n is 5 and m is 6.
[0077] In certain embodiments of a compound of Formula (I), n is 6
and m is 2; n is 6 and m is 3; n is 6 and m is 4; n is 6 and m is
5; and in certain embodiments, n is 6 and m is 6.
[0078] In certain embodiments of a compound of Formula (I), R.sup.1
is chosen from methyl and ethyl.
[0079] In certain embodiments of a compound of Formula (I), R.sup.1
is methyl.
[0080] In certain embodiments of a compound of Formula (I), R.sup.1
is ethyl.
[0081] In certain embodiments of a compound of Formula (I), R.sup.1
is n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl,
pentyl-2-yl, 2-methylbutyl, isopentyl, 3-methylbutan-2-yl,
neopentyl, tert-pentyl, n-hexyl, hexan-2-yl, 2-methylpentyl,
3-methylpentuyl, 4-methylpentyl, 3-methylpentan-2-yl,
4-methylpentan-2-yl, 2,3-dimethylbutyl, and in certain embodiments,
3,3-dimethylbutyl.
[0082] In certain embodiments of a compound of Formula (I), R.sup.1
is
##STR00018##
[0083] In certain embodiments of a compound of Formula (I), R.sup.1
is
##STR00019##
and m is chosen from 2 and 3.
[0084] In certain embodiments of a compound of Formula (I), R.sup.1
is
##STR00020##
and m is chosen from 4, 5, and 6.
[0085] In certain embodiments of a compound of Formula (I), the
compound is a pharmaceutically acceptable salt.
[0086] In certain embodiments of a compound of Formula (I), the
compound is the hydrochloride salt.
[0087] In certain embodiments, a compound of Formula (I) has the
structure of Formula (Ia):
##STR00021##
or a pharmaceutically acceptable salt thereof, wherein n is an
integer from 2 to 6; and R.sup.1 is C.sub.1-6 alkyl.
[0088] In certain embodiments of a compound of Formula (Ia), n is
2. In certain embodiments of a compound of Formula (Ia) where n is
2, the compound is chosen from: [0089] methyl
(2-morpholinoethyl)fumarate; [0090] ethyl(morpholinoethyl)fumarate;
[0091] (morpholinoethyl)propyl fumarate; [0092]
isopropyl(2-morpholinoethyl)fumarate; [0093] butyl
(2-morpholinoethyl)fumarate; [0094] sec-butyl
(2-morpholinoethyl)fumarate; [0095]
isobutyl(2-morpholinoethyl)fumarate; [0096] tert-butyl
(2-morpholinoethyl)fumarate; [0097] (2-morpholinoethyl)pentyl
fumarate; [0098] (2-morpholinoethyl)pentyl-2-yl fumarate; [0099]
2-methylbutyl(2-morpholinoethyl)fumarate; [0100]
isopentyl(2-morpholinoethyl)fumarate; [0101] 3-methylbutan-2-yl
(2-morpholinoethyl)fumarate; [0102] 2-morpholinoethyl neopentyl
fumarate; [0103] 2-morpholinoethyl tert-pentyl fumarate; [0104]
hexyl(2-morpholinoethyl)fumarate; [0105]
hexan-2-yl(2-morpholinoethyl)fumarate; [0106]
2-methylpentyl(2-morpholinoethyl)fumarate; [0107]
3-methylpentyl(2-morpholinoethyl)fumarate; [0108]
4-methylpentyl(2-morpholinoethyl)fumarate; [0109]
3-methylpentan-2-yl(2-morpholinoethyl)fumarate; [0110]
4-methylpentan-2-yl(2-morpholinoethyl)fumarate; [0111]
2,3-dimethylbutyl(2-morpholinoethyl)fumarate; and [0112]
3,3-dimethylbutyl(2-morpholinoethyl)fumarate; or a pharmaceutically
acceptable salt of any of the foregoing.
[0113] In certain embodiments of a compound of Formula (Ia), n is
3. In certain embodiments of a compound of Formula (Ia) where n is
3, the compound is chosen from: [0114] methyl
(3-morpholinopropyl)fumarate; [0115]
ethyl(3-morpholinopropyl)fumarate; [0116]
(3-morpholinopropyl)propyl fumarate; [0117]
isopropyl(3-morpholinopropyl)fumarate; [0118] butyl
(3-morpholinopropyl)fumarate; [0119] sec-butyl
(3-morpholinopropyl)fumarate; [0120]
isobutyl(3-morpholinopropyl)fumarate; [0121] tert-butyl
(3-morpholinopropyl)fumarate; [0122] (3-morpholinopropyl)pentyl
fumarate; [0123] (3-morpholinopropyl)pentyl-2-yl fumarate; [0124]
2-methylbutyl(3-morpholinopropyl)fumarate; [0125]
isopentyl(3-morpholinopropyl)fumarate; [0126]
3-methylbutan-2-yl(3-morpholinopropyl)fumarate; [0127]
3-morpholinopropyl neopentyl fumarate; [0128] 3-morpholinopropyl
tert-pentyl fumarate; [0129] hexyl(3-morpholinopropyl)fumarate;
[0130] hexan-2-yl(3-morpholinopropyl)fumarate; [0131]
2-methylpentyl(3-morpholinopropyl)fumarate; [0132]
3-methylpentyl(3-morpholinopropyl)fumarate; [0133]
4-methylpentyl(3-morpholinopropyl)fumarate; [0134]
3-methylpentan-2-yl(3-morpholinopropyl)fumarate; [0135]
4-methylpentan-2-yl(3-morpholinopropyl)fumarate; [0136]
2,3-dimethylbutyl(3-morpholinopropyl)fumarate; and [0137]
3,3-dimethylbutyl(3-morpholinopropyl)fumarate; or a
pharmaceutically acceptable salt of any of the foregoing.
[0138] In certain embodiments of a compound of Formula (Ia), n is
4. In certain embodiments of a compound of Formula (Ia) where n is
4, the compound is chosen from: [0139] methyl
(4-morpholinobutyl)fumarate; [0140]
ethyl(4-morpholinobutyl)fumarate; [0141] (4-morpholinobutyl)propyl
fumarate; [0142] isopropyl(4-morpholinobutyl)fumarate; [0143] butyl
(4-morpholinobutyl)fumarate; [0144] sec-butyl
(4-morpholinobutyl)fumarate; [0145]
isobutyl(4-morpholinobutyl)fumarate; [0146] tert-butyl
(4-morpholinobutyl)fumarate; [0147] (4-morpholinobutyl)pentyl
fumarate; [0148] (4-morpholinobutyl)pentyl-2-yl fumarate; [0149]
2-methylbutyl(4-morpholinobutyl)fumarate; [0150]
isopentyl(4-morpholinobutyl)fumarate; [0151] 3-methylbutan-2-yl
(4-morpholinobutyl)fumarate; [0152] 4-morpholinobutyl neopentyl
fumarate; [0153] 4-morpholinobutyl tert-pentyl fumarate; [0154]
hexyl(4-morpholinobutyl)fumarate; [0155] hexan-2-yl
(4-morpholinobutyl)fumarate; [0156]
2-methylpentyl(4-morpholinobutyl)fumarate; [0157]
3-methylpentyl(4-morpholinobutyl)fumarate; [0158]
4-methylpentyl(4-morpholinobutyl)fumarate; [0159]
3-methylpentan-2-yl (4-morpholinobutyl)fumarate; [0160]
4-methylpentan-2-yl (4-morpholinobutyl)fumarate; [0161]
2,3-dimethylbutyl(4-morpholinobutyl)fumarate; and [0162]
3,3-dimethylbutyl(4-morpholinobutyl)fumarate; or a pharmaceutically
acceptable salt of any of the foregoing.
[0163] In certain embodiments of a compound of Formula (Ia), n is
5. In certain embodiments of a compound of Formula (Ia) where n is
5, the compound is chosen from: [0164] methyl
(5-morpholinopentyl)fumarate; [0165]
ethyl(5-morpholinopentyl)fumarate; [0166]
(5-morpholinopentyl)propyl fumarate; [0167]
isopropyl(5-morpholinopentyl)fumarate; [0168] butyl
(5-morpholinopentyl)fumarate; [0169] sec-butyl
(5-morpholinopentyl)fumarate; [0170]
isobutyl(5-morpholinopentyl)fumarate; [0171] tert-butyl
(5-morpholinopentyl)fumarate; [0172] (5-morpholinopentyl)pentyl
fumarate; [0173] (5-morpholinopentyl)pentyl-2-yl fumarate; [0174]
2-methylbutyl(5-morpholinopentyl)fumarate; [0175]
isopentyl(5-morpholinopentyl)fumarate; [0176] 3-methylbutan-2-yl
(5-morpholinopentyl)fumarate; [0177] 5-morpholinopentyl neopentyl
fumarate; [0178] 5-morpholinopentyl tert-pentyl fumarate; [0179]
hexyl(5-morpholinopentyl)fumarate; [0180] hexan-2-yl
(5-morpholinopentyl)fumarate; [0181]
2-methylpentyl(5-morpholinopentyl)fumarate; [0182]
3-methylpentyl(5-morpholinopentyl)fumarate; [0183]
4-methylpentyl(5-morpholinopentyl)fumarate; [0184]
3-methylpentan-2-yl (5-morpholinopentyl)fumarate; [0185]
4-methylpentan-2-yl (5-morpholinopentyl)fumarate; [0186]
2,3-dimethylbutyl(5-morpholinopentyl)fumarate; and [0187]
3,3-dimethylbutyl(5-morpholinopentyl)fumarate; or a
pharmaceutically acceptable salt of any of the foregoing.
[0188] In certain embodiments of a compound of Formula (Ia), n is
6. In certain embodiments of a compound of Formula (Ia) where n is
6, the compound is chosen from: [0189] methyl
(6-morpholinohexyl)fumarate; [0190]
ethyl(6-morpholinohexyl)fumarate; [0191] (6-morpholinohexyl)propyl
fumarate; [0192] isopropyl(6-morpholinohexyl)fumarate; [0193] butyl
(6-morpholinohexyl)fumarate; [0194] sec-butyl
(6-morpholinohexyl)fumarate; [0195]
isobutyl(6-morpholinohexyl)fumarate; [0196] tert-butyl
(6-morpholinohexyl)fumarate; [0197] (6-morpholinohexyl)pentyl
fumarate; [0198] (6-morpholinohexyl)pentyl-2-yl fumarate; [0199]
2-methylbutyl(6-morpholinohexyl)fumarate; [0200]
isopentyl(6-morpholinohexyl)fumarate; [0201] 3-methylbutan-2-yl
(6-morpholinohexyl)fumarate; [0202] 6-morpholinohexyl neopentyl
fumarate; [0203] 6-morpholinohexyl tert-pentyl fumarate; [0204]
hexyl(6-morpholinohexyl)fumarate; [0205] hexan-2-yl
(6-morpholinohexyl)fumarate; [0206]
2-methylpentyl(6-morpholinohexyl)fumarate; [0207]
3-methylpentyl(6-morpholinohexyl)fumarate; [0208]
4-methylpentyl(6-morpholinohexyl)fumarate; [0209]
3-methylpentan-2-yl (6-morpholinohexyl)fumarate; [0210]
4-methylpentan-2-yl (6-morpholinohexyl)fumarate; [0211]
2,3-dimethylbutyl(6-morpholinohexyl)fumarate; and [0212]
3,3-dimethylbutyl(6-morpholinohexyl)fumarate; or a pharmaceutically
acceptable salt of any of the foregoing.
[0213] In certain embodiments of compounds of Formula (Ia), the
compound is chosen from: [0214] methyl (morpholinoethyl)fumarate;
[0215] ethyl(2-morpholinoethyl)fumarate; and [0216]
propyl(morpholinoethyl)fumarate; or a pharmaceutically acceptable
salt of any of the foregoing.
[0217] In certain embodiments, any one or more of the compounds of
Formula (Ia) provided in the preceding paragraphs is a
hydrochloride salt.
[0218] In certain embodiments, a compound of Formula (I) has the
structure of Formula (Ib):
##STR00022##
or a pharmaceutically acceptable salt thereof, wherein n is an
integer from 2 to 6; and m is an integer from 2 to 6.
[0219] In certain embodiments of a compound of Formula (Ib), n is
2. In certain embodiments of a compound of Formula (Ib) where n is
2, the compound is chosen from: [0220]
bis(2-morpholinoethyl)fumarate; [0221] 2-morpholinoethyl
(3-morpholinopropyl)fumarate; [0222] 4-morpholinobutyl
(2-morpholinoethyl)fumarate; [0223] 2-morpholinoethyl
(5-morpholinopentyl)fumarate; and [0224] 2-morpholinoethyl
(6-morpholinohexyl)fumarate; or a pharmaceutically acceptable salt
of any of the foregoing.
[0225] In certain embodiments of a compound of Formula (Ib), n is
3. In certain embodiments of a compound of Formula (Ib) where n is
3, the compound is chosen from: [0226] 2-morpholinoethyl
(3-morpholinopropyl)fumarate; [0227]
bis(3-morpholinopropyl)fumarate; [0228] 4-morpholinobutyl
(3-morpholinopropyl)fumarate; [0229] 5-morpholinopentyl
(3-morpholinopropyl)fumarate; and [0230]
6-morpholinohexyl(3-morpholinopropyl)fumarate; or a
pharmaceutically acceptable salt of any of the foregoing.
[0231] In certain embodiments of a compound of Formula (Ib), n is
4. In certain embodiments of a compound of Formula (Ib) where n is
4, the compound is chosen from: [0232] 4-morpholinobutyl
(2-morpholinoethyl)fumarate; [0233] 4-morpholinobutyl
(3-morpholinopropyl)fumarate; [0234]
bis(4-morpholinobutyl)fumarate; [0235] 4-morpholinobutyl
(5-morpholinopentyl)fumarate; and [0236] 4-morpholinobutyl
(6-morpholinohexyl)fumarate; or a pharmaceutically acceptable salt
of any of the foregoing.
[0237] In certain embodiments of a compound of Formula (Ib), n is
5. In certain embodiments of a compound of Formula (Ib) where n is
5, the compound is chosen from: [0238] 2-morpholinoethyl
(5-morpholinopentyl)fumarate; [0239] 5-morpholinopentyl
(3-morpholinopropyl)fumarate; [0240] 4-morpholinobutyl
(5-morpholinopentyl)fumarate; [0241]
bis(5-morpholinopentyl)fumarate; and [0242]
6-morpholinohexyl(5-morpholinopentyl)fumarate; or a
pharmaceutically acceptable salt of any of the foregoing.
[0243] In certain embodiments of a compound of Formula (Ib), n is
6. In certain embodiments of a compound of Formula (Ib) where n is
6, the compound is chosen from: [0244]
2-morpholinoethyl(6-morpholinohexyl)fumarate; [0245]
6-morpholinohexyl(3-morpholinopropyl)fumarate; [0246]
4-morpholinobutyl (6-morpholinohexyl)fumarate; [0247]
6-morpholinohexyl(5-morpholinopentyl)fumarate; and [0248]
bis(6-morpholinohexyl)fumarate; or a pharmaceutically acceptable
salt of any of the foregoing.
[0249] In certain embodiments of compounds of Formula (Ib), the
compound is chosen from: [0250]
2-morpholinoethyl(morpholinopropyl)fumarate; and [0251]
bis(2-morpholinoethyl)fumarate; or a pharmaceutically acceptable
salt of any of the foregoing.
[0252] In certain embodiments, any one or more of the compounds of
Formula (Ib) provided in the preceding paragraphs is a
hydrochloride salt.
[0253] Compounds of Formula (I) may be pharmacologically active or
may be metabolized in vivo to produce metabolites that are
pharmacologically active.
[0254] Compounds provided by the present disclosure include
compounds of Formula (II). Compounds of Formula (II) may optionally
be produced by in vivo metabolism of the corresponding compound of
Formula (I), i.e., by cleavage of the corresponding R.sup.1 moiety.
Alternatively, a compound of Formula (II) may be administered
directly to a patient, for example by placing the compound in a
pharmaceutical preparation or dosage form that is administered to
the patient. Thus, the Formula (II) compounds are themselves
pharmacologically active and require no further metabolism to
become pharmacologically active.
[0255] Accordingly, certain embodiments provide a compound of
Formula (II):
##STR00023##
or a pharmaceutically acceptable salt thereof, wherein n is an
integer from 2 to 6.
[0256] Certain embodiments provide a pharmaceutical composition
comprising a pharmaceutically acceptable vehicle and a
therapeutically effective amount of a compound of Formula (II):
##STR00024##
or a pharmaceutically acceptable salt thereof, wherein n is an
integer from 2 to 6.
[0257] In certain embodiments of a compound of Formula (II), n is
2, n is 3, n is 4, n is 5, and in certain embodiments, n is 6.
[0258] In certain embodiments of a compound of Formula (II), the
compound is a pharmaceutically acceptable salt.
[0259] In certain embodiments of a compound of Formula (II), the
compound is the hydrochloride salt.
[0260] In certain embodiments of a compound of Formula (II), the
compound is chosen from: [0261]
(E)-4-(2-morpholinoethoxy)-4-oxobut-2-enoic acid; [0262]
(E)-4-(3-morpholinopropoxy)-4-oxobut-2-enoic acid; [0263]
(E)-4-(4-morpholinobutoxy)-4-oxobut-2-enoic acid; [0264]
(E)-4-(5-morpholinopentoxy)-4-oxobut-2-enoic acid; and [0265]
(E)-4-(6-morpholinohexoxy)-4-oxobut-2-enoic acid; or a
pharmaceutically acceptable salt of any of the foregoing.
[0266] In a specific aspect, provided here are compounds according
to Formula (I):
##STR00025##
[0267] or a pharmaceutically acceptable salt thereof;
wherein:
[0268] n is an integer from 2 to 6;
[0269] R.sup.1 is selected from H, methyl, ethyl, C.sub.3-6 alkyl,
and
##STR00026##
and
[0270] m is an integer from 2 to 6;
[0271] provided that
[0272] i) when n is 2, and R.sup.1 is
##STR00027##
[0273] then m is 3, 4, 5, or 6; and
[0274] ii) when R.sup.1 is H, then n is 4, 5, or 6.
[0275] In one embodiment, with respect to the compounds of Formula
(I), the compound is a compound according to Formula (IIa), (IIb),
(IIc), (IId), or (IIe):
##STR00028## [0276] or a pharmaceutically acceptable salt thereof;
and wherein R.sup.1 is as described for Formula (I).
[0277] In one embodiment, with respect to the compounds of Formula
(I), (IIa), (IIb), (IIc), (IId), or (IIe), R.sup.1 is methyl,
ethyl, or C.sub.3-6 alkyl. In another embodiment, R.sup.1 is
methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl,
n-pentyl, pentyl-2-yl, 2-methylbutyl, isopentyl,
3-methylbutan-2-yl, neopentyl, tert-pentyl, n-hexyl, hexan-2-yl,
2-methylpentyl, 3-methylpentuyl, 4-methylpentyl,
3-methylpentan-2-yl, 4-methylpentan-2-yl, 2,3-dimethylbutyl, or
3,3-dimethylbutyl. In a particular embodiment, R.sup.1 is
methyl.
[0278] In one embodiment, with respect to the compounds of Formula
(I), R.sup.1 is
##STR00029## [0279] n is 2; and m is 3, 4, 5, or 6.
[0280] In another embodiment, with respect to the compounds of
Formula (I), R.sup.1 is
##STR00030##
and
[0281] n is 3, 4, 5, or 6; and m is 2, 3, 4, 5, or 6.
[0282] In another embodiment, with respect to the compounds of
Formula (I), R.sup.1 is H; and n is 4, 5, or 6.
[0283] In another embodiment, with respect to the compounds of
Formula (I), the compound is a compound according to Formula
(IIIa), (IIIb), (IIIc), (IIId), or (IIIe):
##STR00031##
[0284] or a pharmaceutically acceptable salt thereof.
[0285] In another embodiment, with respect to the compounds of
Formula (I), the compound is a compound according to Formula (IVc),
(IVd), or (IVe):
##STR00032##
[0286] or a pharmaceutically acceptable salt thereof.
[0287] In another embodiment, with respect to the compounds of
Formula (I), (IIa)-(IIe), (IIIa)-(IIIe), and (IVc)-(IVe), the
compound is a pharmaceutically acceptable salt. In a particular
embodiment, the compound is a HCl salt.
[0288] In a specific embodiment, with respect to the compounds of
Formula (I), the compound is any one of the compounds listed in
Table 1.
[0289] In a particular embodiment, with respect to the compounds of
Formula (I), the compound according to Formula (IIIc):
##STR00033##
[0290] or a pharmaceutically acceptable salt thereof.
[0291] In a more particular embodiment, with respect to the
compounds of Formula (I), the compound is a HCl salt of (IIIc).
[0292] In another specific aspect, provided here are pharmaceutical
compositions comprising a pharmaceutically acceptable vehicle and a
therapeutically effective amount of a compound of Formula (I):
##STR00034##
[0293] or a pharmaceutically acceptable salt thereof;
wherein:
[0294] n is an integer from 2 to 6;
[0295] R.sup.1 is selected from H, methyl, ethyl, C.sub.3-6 alkyl,
and
##STR00035##
and
[0296] m is an integer from 2 to 6.
[0297] In one embodiment, with respect to the pharmaceutical
compositions, the compound is a compound according to Formula
(IIa), (IIb), (IIc), (IId), or (IIe) or a pharmaceutically
acceptable salt thereof; and wherein R.sup.1 is as described for
Formula (I); and Formulae (IIa)-(IIe) are as depicted above.
[0298] In one embodiment, with respect to the pharmaceutical
compositions of Formula (I), (IIa), (IIb), (IIc), (IId), or (IIe),
R.sup.1 is methyl, ethyl, or C.sub.3-6 alkyl. In another
embodiment, R.sup.1 is methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, tert-butyl, n-pentyl, pentyl-2-yl, 2-methylbutyl,
isopentyl, 3-methylbutan-2-yl, neopentyl, tert-pentyl, n-hexyl,
hexan-2-yl, 2-methylpentyl, 3-methylpentuyl, 4-methylpentyl,
3-methylpentan-2-yl, 4-methylpentan-2-yl, 2,3-dimethylbutyl, or
3,3-dimethylbutyl. In a particular embodiment, R.sup.1 is
methyl.
[0299] In one embodiment, with respect to the pharmaceutical
compositions of Formula (I), R.sup.1 is
##STR00036## [0300] and m is 2, 3, 4, 5, or 6.
[0301] In another embodiment, with respect to the pharmaceutical
compositions of Formula (I) (IIa), (IIb), (IIc), (IId), or (IIe),
R.sup.1 is H. In one embodiment, R.sup.1 is H; and n is 2. In
another embodiment, R.sup.1 is H; and n is 3. In a yet another
embodiment, R.sup.1 is H; and n is 4, 5, or 6.
[0302] In another embodiment, with respect to the pharmaceutical
compositions of Formula (I), the compound is a compound according
to Formula (IIIa), (IIIb), (IIIc), (IIId), or (IIIe), or a
pharmaceutically acceptable salt thereof; and Formulae are as
depicted above.
[0303] In another embodiment, with respect to the pharmaceutical
compositions of Formula (I), (IIa)-(IIe), and (IIIa)-(IIIe), the
compound is a pharmaceutically acceptable salt. In a particular
embodiment, the compound is a HCl salt.
[0304] In a specific embodiment, with respect to the pharmaceutical
compositions of Formula (I), the compound is any one of the
compounds listed in Table 1.
[0305] In a particular embodiment, with respect to the
pharmaceutical compositions of Formula (I), the compound according
to Formula (IIIc):
##STR00037##
[0306] or a pharmaceutically acceptable salt thereof.
[0307] In a more particular embodiment, with respect to the
pharmaceutical compositions of Formula (I), the compound is a HCl
salt of (IIIc).
[0308] In another specific aspect, provided here are method for
preventing or treating in a mammal in need thereof a disease or
condition which comprises administering to the mammal an effective
disease-treating or condition-treating amount of a compound
according to Formula (I):
##STR00038##
[0309] or a pharmaceutically acceptable salt thereof;
wherein:
[0310] n is an integer from 2 to 6;
[0311] R.sup.1 is selected from H, methyl, ethyl, C.sub.3-6 alkyl,
and
##STR00039##
and
[0312] m is an integer from 2 to 6.
[0313] In one embodiment, with respect to the methods, the compound
is a compound according to Formula (IIa), (IIb), (IIc), (IId), or
(IIe) or a pharmaceutically acceptable salt thereof; and R.sup.1 is
as described for Formula (I); and Formulae (IIa)-(IIe) are as
depicted above.
[0314] In one embodiment, with respect to the methods, the compound
is a compound according to Formula (I), (IIa), (IIb), (IIc), (IId),
or (IIe), and R.sup.1 is methyl, ethyl, or C.sub.3-6 alkyl. In
another embodiment, R.sup.1 is methyl, ethyl, n-propyl, isopropyl,
n-butyl, isobutyl, tert-butyl, n-pentyl, pentyl-2-yl,
2-methylbutyl, isopentyl, 3-methylbutan-2-yl, neopentyl,
tert-pentyl, n-hexyl, hexan-2-yl, 2-methylpentyl, 3-methylpentuyl,
4-methylpentyl, 3-methylpentan-2-yl, 4-methylpentan-2-yl,
2,3-dimethylbutyl, or 3,3-dimethylbutyl. In a particular
embodiment, R.sup.1 is methyl.
[0315] In one embodiment, with respect to the methods, the compound
is a compound according to Formula (I), R.sup.1 is
##STR00040## [0316] and m is 2, 3, 4, 5, or 6.
[0317] In another embodiment, with respect to the methods, the
compound is a compound according to Formula (I) (IIa), (IIb),
(IIc), (IId), or (IIe), and R.sup.1 is H. In one embodiment,
R.sup.1 is H; and n is 2. In another embodiment, R.sup.1 is H; and
n is 3. In a yet another embodiment, R.sup.1 is H; and n is 4, 5,
or 6.
[0318] In another embodiment, with respect to the methods, the
compound is a compound according to Formula (I), and the compound
is a compound according to Formula (IIIa), (IIIb), (IIIc), (IIId),
or (IIIe), or a pharmaceutically acceptable salt thereof; and
Formulae (IIIa)-(IIIe) are as depicted above.
[0319] In another embodiment, with respect to the methods, the
compound is a compound according to Formula (I), (IIa)-(IIe), and
(IIIa)-(IIIe), and the compound is a pharmaceutically acceptable
salt. In a particular embodiment, the compound is a HCl salt.
[0320] In a specific embodiment, with respect to the methods, the
compound is any one of the compounds listed in Table 1.
[0321] In a particular embodiment, with respect to the methods, the
compound is according to Formula (IIIc):
##STR00041##
[0322] or a pharmaceutically acceptable salt thereof.
[0323] In a more particular embodiment, with respect to the
methods, the compound is a HCl salt of (IIIc).
[0324] In one embodiment, with respect to the methods, the disease
or condition is selected from a neurodegenerative disease, an
inflammatory disease, and an autoimmune disease. In certain
embodiments, the disease or condition is selected from multiple
sclerosis, psoriasis, irritable bowel disorder, ulcerative colitis,
arthritis, chronic obstructive pulmonary disease, asthma,
Parkinson's disease, Huntington's disease, and amyotrophic lateral
sclerosis.
Synthesis
[0325] Compounds disclosed herein may be obtained via the synthetic
methods illustrated in Schemes 1 through 4. In addition, general
synthetic methods useful in the synthesis of compounds described
herein are available in the art. Starting materials useful for
preparing compounds and intermediates thereof and/or practicing
methods described herein are commercially available or can be
prepared by well-known synthetic methods. The methods presented in
the schemes provided by the present disclosure are illustrative
rather than comprehensive. It will be apparent to those skilled in
the art that many modifications, both to materials and methods, may
be practiced without departing from the scope of the
disclosure.
[0326] Certain of the halo alkyl morpholines useful for preparing
compounds of the disclosure are available from commercial sources.
Non-commercially available halo alkyl morpholines useful for
preparing compounds of the disclosure, and intermediates thereof
may be prepared by well-known synthetic methods such as those
described in Schemes 1 and 2.
[0327] Functionalized 1-halo alkyl morpholines useful for the
preparation of morpholinoalkyl fumarates of compounds of the
disclosure may be prepared according to Scheme 1:
##STR00042##
where X and Y are leaving groups such as halogen and n is as
defined in Formula (I) and Formula (II). In certain embodiments of
Scheme 1, X is chloro and Y is selected from chloro and an
O-acylisourea.
[0328] Chemical activation of the alcohol to the corresponding
chloride as shown in Scheme 1 may be achieved by reaction with
chlorination agents such as thionyl chloride (SOCl.sub.2), oxalyl
chloride (C.sub.2O.sub.2Cl.sub.2), or phosphorous pentachloride
(PCl.sub.5), optionally in the presence of a suitable catalyst such
as N,N-dimethylformamide, and either in substance (absence of
solvent) or in an inert organic solvent such as dichloromethane
(DCM) at an appropriate temperature such as from about 0.degree. C.
to about 70.degree. C. Chemical activation of the alcohol may be
performed in situ and without isolating the activated substrate
prior to the following aminolysis step. Optionally, the activated
alcohol may be isolated and/or purified using methods well known in
the art, i.e. fractional distillation.
[0329] Alternatively, carbodiimide dehydration agents such as
N,N'-diisopropylcarbodiimide (DIC), N,N'-dicyclohexylcarbodiimide
(DCC), or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAC,
EDC), optionally in the presence of a catalytic or stoichiometric
amount of a suitable additive such as 4-(N,N-dimethylaminopyridine)
(DMAP) (Steglich esterification conditions), 1-hydroxybenzotriazole
(HOBt), 1-hydroxy-7-aza-benzotriazole (HOAt), or
N-hydroxysuccinimide (NHS); uronium or phosphonium salts with
non-nucleophilic anions such as
N-[(1H-benzotriazol-1-yl)(dimethylamino)methylene]-N-methylmetanaminium
hexafluorophosphate (HBTU),
N-[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridin-1-ylmethylene]-N-methy-
lmetanaminium hexafluorophosphate N-oxide (HATU),
N-[(1H-benzotriazol-1-yl)(dimethylamino)methylene]-N-methylmetanaminium
tetrafluoroborate (TBTU), or
benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate
(PyBOP), can be employed to form an activated carboxylic acid
derivative. Optionally, organic tertiary bases such as
triethylamine (TEA) or diisopropylethylamine (DIEA) can also be
employed. The formation of the activated carboxylic acid derivative
can take place in an inert solvent such as dichloromethane (DCM),
N,N-dimethylformamide, N-methylpyrrolidone (NMP),
N,N-dimethylacetamide (DMA, DMAc), or mixtures of any of the
foregoing at an appropriate temperature such as from about
0.degree. C. to about 40.degree. C.
[0330] Aminolysis of in situ generated or isolated activated
carboxylic derivatives with the appropriately functionalized amine
derivative (morpholine) (Scheme 2) can take place in the presence
of a suitable base such as an organic tertiary base, i.e.,
triethylamine (TEA), diethylaminoethylamine (DIEA), pyridine, or
mixtures of any of the foregoing, optionally in the presence of
suitable additives such as nucleophilic acylation catalysts, i.e.,
4-(N,N-dimethylaminopyridine) (DMAP), and in the same or other
inert solvent as used for the activation step such as
dichloromethane (DCM), N,N-dimethylformamide, N-methylpyrrolidone
(NMP), N,N-dimethylacetamide (DMA, DMAc), or mixtures of any of the
foregoing, at an appropriate temperature such as from about
0.degree. C. to about 70.degree. C.
[0331] Functionalized 1-hydroxy alkyl morpholines useful for the
preparation of morpholinoalkyl fumarates of the compounds of
disclosure may be also prepared according to Scheme 2:
##STR00043##
where PG is a hydroxyl protecting group; Y is a leaving group such
as chloro or an O-isourea derived radical; and n is as defined in
Formula (I) and Formula (II).
[0332] Certain of the functionalized and activated diols are
commercially available. Methods for introducing hydroxyl protecting
groups (PGs) are well known in the art. Useful protecting groups to
temporarily block the hydroxyl group of functionalized diols
include certain alkyl ethers such as (substituted) benzyl ethers,
tert-butyl ethers, trityl ether, or various silyl ethers such as
tert-butyl dimethylsilyl ether, triisopropylsilyl ether, or
tert-butyldiphenylsilyl ethers.
[0333] Certain protected, functionalized and activated diols are
commercially available. Alternatively, the chemical activation of
the protected and functionalized diols to the corresponding
activated alcohol, i.e., alcohol chloride, may be achieved using
similar reaction procedures and conditions as those described in
Scheme 1 for the activation of functionalized 1-halo alcohols.
[0334] Aminolysis of in situ generated or isolated protected,
functionalized, and activated 1-hydroxy derivatives with morpholine
may take place using similar reaction procedures and conditions as
those described in Scheme 1 for the aminolysis of functionalized,
protected, and activated 1-halo alcohols.
[0335] Orthogonal (or ordered) deprotection of the protected
1-hydroxyacetic acid derivative liberates the corresponding free
hydroxyl group. Deprotection methods, procedures, and practices are
well known in the art.
[0336] In certain embodiments, the protecting group can be an alkyl
group such as a tert-butyl group. Deprotection may be carried out
by contacting a tert-butyl protected functionalized 1-hydroxy
acetamide derivative with an excess of a strong Bronsted acid such
as trifluoroacetic acid (TFA) or hydrogen chloride (HCl) in an
inert solvent such as dichloromethane (DCM), diethyl ether
(Et.sub.2O), 1,4-dioxane, or mixtures of any of the foregoing, at
an appropriate temperature such as from about 0.degree. C. to about
40.degree. C.
[0337] In certain embodiments, the protecting group can be chosen
from an alkyl group such as a benzyl group. When the protecting
group is a benzyl group, deprotection may be carried out by
reacting the functionalized 1-hydroxy acetamide derivative with
gaseous hydrogen (H.sub.2) in the presence of a heterogeneous
catalyst, i.e., 5-10 wt-% palladium on activated carbon (activated
or wet coal), in a solvent such as methanol (MeOH), ethanol (EtOH),
ethyl acetate (EtOAc), or mixtures of any of the foregoing,
optionally in the presence of a small amount of an activator such
as 1 N aq. hydrochloric acid at an appropriate temperature such as
from about 0.degree. C. to about 40.degree. C. and under a hydrogen
atmosphere at a pressure of about 15 psi to about 60 psi.
[0338] Morpholinoalkyl fumarates of Formula (I) and Formula (II)
can be prepared according to Scheme 3:
##STR00044##
where X is a leaving group such as halogen, and R.sup.1 and n are
as defined in Formula (I) or R.sup.1 is hydrogen as defined in
Formula (II). In certain embodiments of Scheme 3, X is chloro and
R.sup.1 is C.sub.1-6 alkyl such as methyl or ethyl.
[0339] Nucleophilic displacement of the monoalkyl fumaric acid with
the functionalized 1-halo alkyl morpholine (Scheme 1) as shown in
Scheme 3 may take place in the presence of an inorganic base such
as an alkali carbonate such as cesium hydrogencarbonate
(CsHCO.sub.3), cesium carbonate (Cs.sub.2CO.sub.3), or potassium
carbonate (K.sub.2CO.sub.3). Optionally, organic tertiary bases
such as triethylamine (TEA), diisopropylethylamine (DIEA), or
amidine; guanidine-based bases such as
1,5-diazabicyclo[4.3.0]non-5-ene (DBN),
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) or
1,1,3,3-tetramethylguanidine (TMG); silver salts such silver(I)
oxide (Ag.sub.2O) or silver(I) carbonate (Ag2CO.sub.3); or other
halide scavengers known in the art can be employed. The
corresponding alkali, tri- and tetraalkylammonium, amidine, or
guanide salts of the monoalkyl fumarate can be generated in situ
or, alternatively, can be prepared separately. The reaction can
take place in an inert solvent such as N,N-dimethylformamide,
N-methylpyrrolidone (NMP), N,N-dimethylacetamide (DMA, DMAc),
dimethylsulfoxide (DMSO), tetrahydrofuran (THF), toluene, or
mixtures of any of the foregoing at an appropriate temperature such
as from about room temperature to about 70.degree. C.
[0340] Morpholinoalkyl fumarates of Formula (I) and Formula (II)
may also be prepared according to Scheme 4:
##STR00045##
where Y is a suitable leaving group such as halogen, an
O-acylisourea, various triazolol esters, or others; and R.sup.1 and
n are as defined herein. In certain embodiments of Scheme 4, Y is
chloro and R.sup.1 is C.sub.1-6 alkyl such as methyl or ethyl for a
compound of Formula (I) or R.sup.1 is hydrogen for a compound of
Formula (II).
[0341] Chemical activation of the carboxylic acid to the
corresponding carboxylic acid chloride as shown in Scheme 4 may be
accomplished by reaction with a chlorination agent such as thionyl
chloride (SOCl.sub.2), oxalyl chloride (C.sub.2O.sub.2Cl.sub.2),
phosphorous pentachloride (PCl.sub.5), or others, optionally in the
presence of a catalyst such as N,N-dimethylformamide, and either in
substance (absence of solvent) or in an inert organic solvent such
as dichloromethane (DCM) at an appropriate temperature such as from
about 0.degree. C. to about 70.degree. C. Chemical activation of
the carboxylic acid as shown in Scheme 4 may be performed in situ
without isolating the activated substrate prior to the subsequent
alcoholysis step. Optionally, the activated carboxylic acid
chloride may be isolated and/or purified using methods well known
in the art, i.e. fractional distillation.
[0342] Alternatively, carbodiimide dehydration agents such as
N,N'-diisopropylcarbodiimide (DIC), N,N'-dicyclohexylcarbodiimide
(DCC), or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAC,
EDC), optionally in the presence of a catalytic or stoichiometric
amount of an additive such as 4-(N,N-dimethylaminopyridine) (DMAP)
(Steglich esterification conditions), 1-hydroxybenzotriazole
(HOBt), 1-hydroxy-7-aza-benzotriazole (HOAt), or
N-hydroxysuccinimide (HOSu); a uronium or phosphonium salt with
non-nucleophilic anions such as
N-[(1H-benzotriazol-1-yl)(dimethylamino)methylene]-N-methylmetanaminium
hexafluorophosphate (HBTU),
N--[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridin-1-ylmethylene]-N-meth-
ylmetanaminium hexafluorophosphate N-oxide (HATU),
N-[(1H-benzotriazol-1-yl)(dimethylamino)methylene]-N-methylmetanaminium
tetrafluoroborate (TBTU), or
benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate
(PyBOP), may be employed to form an activated monoalkyl fumarate
derivative. Optionally, organic tertiary bases such as
triethylamine (TEA) or diethylaminoethylamine (DIEA) can also be
employed. The formation of activated monoalkyl fumarate derivatives
may take place in an inert solvent such as dichloromethane (DCM),
N,N-dimethylformamide, N-methylpyrrolidone (NMP),
N,N-dimethylacetamide (DMA, DMAc), or mixtures of any of the
foregoing at an appropriate temperature such as from about room
temperature to about 70.degree. C.
[0343] Alcoholysis of the activated monoalkyl fumarate derivative
with a functionalized hydroxy alkyl morpholine derivative (Scheme
2) as shown in Scheme 4 may take place in the presence of a base,
for example, an organic tertiary base such as, triethylamine (TEA),
diethylaminoethylamine (DIEA), or pyridine, optionally in the
presence of an additive such as a nucleophilic acylation catalyst,
i.e., 4-(N,N-dimethylaminopyridine) (DMAP) (Steglich esterification
conditions), and in the same or other inert solvent as used for the
activation step such as dichloromethane (DCM),
N,N-dimethylformamide, N-methylpyrrolidone (NMP),
N,N-dimethylacetamide (DMA, DMAc), or mixtures of any of the
foregoing at an appropriate temperature such as from about
0.degree. C. to about 70.degree. C.
Pharmaceutical Compositions
[0344] Pharmaceutical compositions provided by the present
disclosure may comprise a therapeutically effective amount of a
compound of Formula (I) or Formula (II) together with a suitable
amount of one or more pharmaceutically acceptable vehicles so as to
provide a composition for proper administration to a patient.
Suitable pharmaceutical vehicles are described in the art.
[0345] In certain embodiments, a compound of Formula (I) or Formula
(II) may be incorporated into pharmaceutical compositions to be
administered orally. Oral administration of such pharmaceutical
compositions may result in uptake of a compound of Formula (I) or
Formula (II) throughout the intestine and entry of such compound
into the systemic circulation. Such oral compositions may be
prepared in a manner known in the pharmaceutical art and comprise a
compound of Formula (I) or Formula (II) and at least one
pharmaceutically acceptable vehicle. Oral pharmaceutical
compositions may include a therapeutically effective amount of a
compound of Formula (I) or Formula (II) and a suitable amount of a
pharmaceutically acceptable vehicle, so as to provide an
appropriate form for oral administration to a patient.
[0346] Compounds of Formula (I) or Formula (II) may be incorporated
into pharmaceutical compositions to be administered by any
appropriate route of administration including intradermal,
intramuscular, intraperitoneal, intravenous, subcutaneous,
intranasal, epidural, oral, sublingual, intracerebral,
intravaginal, transdermal, rectal, inhalation, or topical. In
certain embodiments, compounds of Formula (I) or Formula (II) may
be incorporated into pharmaceutical compositions to be administered
orally.
[0347] Pharmaceutical compositions comprising a compound of Formula
(I) or Formula (II) and may be manufactured by means of
conventional mixing, dissolving, granulating, dragee-making,
levigating, emulsifying, encapsulating, entrapping, or lyophilizing
processes. Pharmaceutical compositions may be formulated in a
conventional manner using one or more physiologically acceptable
carriers, diluents, excipients, or auxiliaries, which facilitate
processing of compounds of Formula (I), Formula (II), or
crystalline forms thereof and one or more pharmaceutically
acceptable vehicles into formulations that can be used
pharmaceutically. Proper formulation is in part dependent upon the
route of administration chosen. Pharmaceutical compositions
provided by the present disclosure may take the form of solutions,
suspensions, emulsions, tablets, pills, pellets, capsules, capsules
containing liquids, powders, sustained-release formulations,
suppositories, emulsions, aerosols, sprays, suspensions, or any
other form suitable for administration to a patient.
[0348] Pharmaceutical compositions provided by the present
disclosure may be formulated in a unit dosage form. A unit dosage
form refers to a physically discrete unit suitable as a unitary
dose for patients undergoing treatment, with each unit containing a
predetermined quantity of a compound of Formula (I) or Formula (II)
calculated to produce an intended therapeutic effect. A unit dosage
form may be for a single daily dose, for administration 2 times per
day, or one of multiple daily doses, e.g., 3 or more times per day.
When multiple daily doses are used, a unit dosage form may be the
same or different for each dose. One or more dosage forms may
comprise a dose, which may be administered to a patient at a single
point in time or during a time interval.
[0349] Pharmaceutical compositions comprising a compound of Formula
(I) or Formula (II) may be formulated for immediate release.
[0350] In certain embodiments, an oral dosage form provided by the
present disclosure may be a controlled release dosage form.
Controlled delivery technologies can improve the absorption of a
drug in a particular region, or regions, of the gastrointestinal
tract. Controlled drug delivery systems may be designed to deliver
a drug in such a way that the drug level is maintained within a
therapeutically effective window and effective and safe blood
levels are maintained for a period as long as the system continues
to deliver the drug with a particular release profile in the
gastrointestinal tract. Controlled drug delivery may produce
substantially constant blood levels of a drug over a period of time
as compared to fluctuations observed with immediate release dosage
forms. For some drugs, maintaining a constant blood and tissue
concentration of the drug throughout the course of therapy is the
most desirable mode of treatment. Immediate release of drugs may
cause blood levels to peak above a level required to elicit a
desired response, which may waste the drug and/or may cause or
exacerbate toxic side effects. Controlled drug delivery can result
in optimum therapy, and not only can reduce the frequency of
dosing, but may also reduce the severity of side effects. Examples
of controlled release dosage forms include dissolution controlled
systems, diffusion controlled systems, ion exchange resins,
osmotically controlled systems, erodable matrix systems, pH
independent formulations, and gastric retention systems.
[0351] An appropriate oral dosage form for a particular
pharmaceutical composition provided by the present disclosure may
depend, at least in part, on the gastrointestinal absorption
properties of a compound of Formula (I) or Formula (II), the
stability of a compound of Formula (I) or Formula (II) in the
gastrointestinal tract, the pharmacokinetics of a compound of
Formula (I) or Formula (II) and the intended therapeutic profile.
An appropriate controlled release oral dosage form may be selected
for a particular compound of Formula (I) or Formula (II). For
example, gastric retention oral dosage forms may be appropriate for
compounds absorbed primarily from the upper gastrointestinal tract,
and sustained release oral dosage forms may be appropriate for
compounds absorbed primarily from the lower gastrointestinal tract.
Certain compounds are absorbed primarily from the small intestine.
In general, compounds traverse the length of the small intestine in
about 3 to 5 hours. For compounds that are not easily absorbed by
the small intestine or that do not dissolve readily, the window for
active agent absorption in the small intestine may be too short to
provide a desired therapeutic effect.
[0352] In certain embodiments, pharmaceutical compositions provided
by the present disclosure may be practiced with dosage forms
adapted to provide sustained release of a compound of Formula (I)
or Formula (II) upon oral administration. Sustained release oral
dosage forms may be used to release drugs over a prolonged time
period and are useful when it is desired that a drug or drug form
be delivered to the lower gastrointestinal tract. Sustained release
oral dosage forms include any oral dosage form that maintains
therapeutic concentrations of a drug in a biological fluid such as
the plasma, blood, cerebrospinal fluid, or in a tissue or organ for
a prolonged time period. Sustained release oral dosage forms
include diffusion-controlled systems such as reservoir devices and
matrix devices, dissolution-controlled systems, osmotic systems,
and erosion-controlled systems. Sustained release oral dosage forms
and methods of preparing the same are well known in the art.
[0353] An appropriate dose of a compound of Formula (I) or Formula
(II), or pharmaceutical composition comprising a compound of
Formula (I) or Formula (II), may be determined according to any one
of several well-established protocols. For example, animal studies,
such as studies using mice, rats, dogs, and/or monkeys, may be used
to determine an appropriate dose of a pharmaceutical compound.
Results from animal studies may be extrapolated to determine
appropriate doses for use in other species, such as for example,
humans.
Uses
[0354] Compounds of Formula (I) are derivatives of monoalkyl
hydrogen fumarates. Compounds of Formula (II) are morpholinoalkyl
esters of fumaric acid. Thus, compounds of Formula (I) or Formula
(II) and pharmaceutical compositions thereof may be administered to
a patient suffering from any disease including a disorder,
condition, or symptom for which monoalkyl hydrogen fumarates and/or
fumaric acid esters are known or hereafter discovered to be
therapeutically effective. Indications for which methyl hydrogen
fumarate (MHF) has been prescribed, and hence for which a compound
of Formula (I) or Formula (II), or pharmaceutical compositions
thereof are also expected to be effective, include psoriasis. Other
indications for which compounds of Formula (I) or Formula (II) may
be therapeutically effective include multiple sclerosis, irritable
bowel disorder, ulcerative colitis, arthritis, chronic obstructive
pulmonary disease, asthma, Parkinson's disease, Huntington's
disease, and amyotrophic lateral sclerosis.
[0355] Methods of treating a disease in a patient provided by the
present disclosure comprise administering to a patient in need of
such treatment a therapeutically effective amount or dose of a
compound of Formula (I) or Formula (II). Compounds of Formula (I)
or Formula (II) or pharmaceutical compositions thereof may provide
therapeutic or prophylactic plasma and/or blood concentrations of
fumarate following administration to a patient.
[0356] Morpholinoalkyl fumarates of Formula (I) or Formula (II) may
be included in a pharmaceutical composition and/or dosage form
adapted for oral administration, although compounds of Formula (I)
or Formula (II) may also be administered by any other appropriate
route, such as for example, by injection, infusion, inhalation,
transdermally, or absorption through epithelial or mucosal
membranes (e.g., oral, rectal, and/or intestinal mucosa).
[0357] Morpholinoalkyl fumarates of Formula (I) or Formula (II) may
be administered in an amount and using a dosing schedule as
appropriate for treatment of a particular disease. Daily doses of
compounds of Formula (I) or Formula (II) may range from about 0.01
mg/kg to about 50 mg/kg, from about 0.1 mg/kg to about 50 mg/kg,
from about 1 mg/kg to about 50 mg/kg, and in certain embodiments,
from about 5 mg/kg to about 25 mg/kg. In certain embodiments,
compounds of Formula (I) or Formula (II) may be administered at a
dose over time from about 1 mg to about 5 g per day, from about 10
mg to about 4 g per day, and in certain embodiments from about 20
mg to about 2 g per day. An appropriate dose of a compound of
Formula (I) or Formula (II) may be determined based on several
factors, including, for example, the body weight and/or condition
of the patient being treated, the severity of the disease being
treated, the incidence and/or severity of side effects, the manner
of administration, and the judgment of the prescribing physician.
Appropriate dose ranges may be determined by methods known to those
skilled in the art.
[0358] Compounds of Formula (I) or Formula (II) may be assayed in
vitro and in vivo for the desired therapeutic or prophylactic
activity prior to use in humans. In vivo assays, for example using
appropriate animal models, may also be used to determine whether
administration of a compound of Formula (I) or Formula (II) is
therapeutically effective.
[0359] In certain embodiments, a therapeutically effective dose of
a compound of Formula (I) or Formula (II) may provide therapeutic
benefit without causing substantial toxicity including adverse side
effects. Toxicity of compounds of Formula (I) or Formula (II)
and/or metabolites thereof may be determined using standard
pharmaceutical procedures and may be ascertained by those skilled
in the art. The dose ratio between toxic and therapeutic effect is
the therapeutic index. A dose of a compound of Formula (I) or
Formula (II) may be within a range capable of establishing and
maintaining a therapeutically effective circulating plasma and/or
blood concentration of a compound of Formula (I) or Formula (II)
that exhibits little or no toxicity.
[0360] Compounds of Formula (I) or Formula (II) may be used to
treat diseases, disorders, conditions, and symptoms of any of the
foregoing for which alkyl hydrogen fumarates, such as MHF, are
known to provide or are later found to provide therapeutic benefit.
MHF is known to be effective in treating psoriasis, multiple
sclerosis, an inflammatory bowel disease, asthma, chronic
obstructive pulmonary disease, and arthritis. Hence, compounds of
Formula (I) or Formula (II) may also be used to treat any of these
diseases and disorders. The underlying etiology of any of the
foregoing diseases being treated may have a multiplicity of
origins. Further, in certain embodiments, a therapeutically
effective amount of one or more compounds of Formula (I) or Formula
(II) may be administered to a patient, such as a human, as a
preventative measure against various diseases or disorders. Thus, a
therapeutically effective amount of one or more compounds of
Formula (I) or Formula (II) may be administered as a preventative
measure to a patient having a predisposition for and/or history of
immunological, autoimmune, and/or inflammatory diseases including
psoriasis, arthritis, asthma, and chronic obstructive pulmonary
disease; cardiac insufficiency including left ventricular
insufficiency, myocardial infarction, and angina pectoris;
mitochondrial and neurodegenerative diseases such as Parkinson's
disease, Alzheimer's disease, Huntington's disease, amyotrophic
lateral sclerosis, retinopathia pigmentosa, and mitochondrial
encephalomyopathy; transplantation rejection; autoimmune diseases
such as multiple sclerosis; ischemia and reperfusion injury;
AGE-induced genome damage; inflammatory bowel diseases such as
Crohn's disease, irritable bowel disorder, and ulcerative colitis;
and NF-.kappa.B mediated diseases.
Psoriasis
[0361] Psoriasis is characterized by hyperkeratosis and thickening
of the epidermis as well as by increased vascularity and
infiltration of inflammatory cells in the dermis. Psoriasis
vulgaris manifests as silvery, scaly, erythematous plaques on
typically the scalp, elbows, knees, and buttocks. Guttate psoriasis
occurs as tear-drop size lesions.
[0362] Fumaric acid esters are recognized for the treatment of
psoriasis and dimethyl fumarate is approved for the systemic
treatment of psoriasis in Germany (Mrowietz and Asadullah, Trends
Mol Med 2005, 11(1), 43-48; and Mrowietz et al., Br J Dermatology
1999, 141, 424-429).
[0363] Efficacy of compounds of Formula (I) or Formula (II) for
treating psoriasis can be determined using animal models and in
clinical trials.
Inflammatory Arthritis
[0364] Inflammatory arthritis includes diseases such as rheumatoid
arthritis, juvenile rheumatoid arthritis (juvenile idiopathic
arthritis), psoriatic arthritis, and ankylosing spondylitis produce
joint inflammation. The pathogenesis of immune-mediated
inflammatory diseases including inflammatory arthritis is believed
to involve TNF and NF-.kappa.B signaling pathways (Tracey et al.,
Pharmacology & Therapeutics 2008, 117, 244-279). DMF has been
shown to inhibit TNF and inflammatory diseases including
inflammatory arthritis, which are believed to involve TNF and
NK-.kappa.B signaling, and therefore may be useful in treating
inflammatory arthritis (Lowewe et al., J Immunology 2002, 168,
4781-4787).
[0365] The efficacy of compounds of Formula (I) or Formula (II) for
treating inflammatory arthritis can be determined using animal
models and in clinical trials.
Multiple Sclerosis
[0366] Multiple sclerosis (MS) is an inflammatory autoimmune
disease of the central nervous system caused by an autoimmune
attack against the insulating axonal myelin sheaths of the central
nervous system. Demyelination leads to the breakdown of conduction
and to severe disease with destruction of local axons and
irreversible neuronal cell death. The symptoms of MS are highly
varied with each individual patient exhibiting a particular pattern
of motor, sensible, and sensory disturbances. MS is typified
pathologically by multiple inflammatory foci, plaques of
demyelination, gliosis, and axonal pathology within the brain and
spinal cord, all of which contribute to the clinical manifestations
of neurological disability (see e.g., Wingerchuk, Lab Invest 2001,
81, 263-281; and Virley, NeuroRx 2005, 2(4), 638-649). Although the
causal events that precipitate MS are not fully understood,
evidence implicates an autoimmune etiology together with
environmental factors, as well as specific genetic predispositions.
Functional impairment, disability, and handicap are expressed as
paralysis, sensory and octintive disturbances, spasticity, tremor,
a lack of coordination, and visual impairment, which impact the
quality of life of the individual. The clinical course of MS can
vary from individual to individual, but invariability of the
disease can be categorized in three forms: relapsing-remitting,
secondary progressive, and primary progressive.
[0367] Studies support the efficacy of fumaric acid esters for
treating MS, which are presently undergoing phase II clinical
testing (Schimrigk et al., Eur J Neurology 2006, 13, 604-610; and
Wakkee and Thio, Current Opinion Investigational Drugs 2007, 8(11),
955-962).
[0368] Assessment of MS treatment efficacy in clinical trials can
be accomplished using tools such as the Expanded Disability Status
Scale and the MS Functional as well as magnetic resonance imaging
lesion load, biomarkers, and self-reported quality of life. Animal
models of MS shown to be useful to identify and validate potential
therapeutics include experimental autoimmune/allergic
encephalomyelitis (EAE) rodent models that simulate the clinical
and pathological manifestations of MS and nonhuman primate EAE
models.
Inflammatory Bowel Disease (Crohn's Disease, Ulcerative
Colitis)
[0369] Inflammatory bowel disease (IBD) is a group of inflammatory
conditions of the large intestine and in some cases, the small
intestine that includes Crohn's disease and ulcerative colitis.
Crohn's disease, which is characterized by areas of inflammation
with areas of normal lining in between, can affect any part of the
gastrointestinal tract from the mouth to the anus. The main
gastrointestinal symptoms are abdominal pain, diarrhea,
constipation, vomiting, weight loss, and/or weight gain. Crohn's
disease can also cause skin rashes, arthritis, and inflammation of
the eye. Ulcerative colitis is characterized by ulcers or open
sores in the large intestine or colon. The main symptom of
ulcerative colitis is typically constant diarrhea with mixed blood
of gradual onset. Other types of intestinal bowel disease include
collagenous colitis, lymphocytic colitis, ischemic colitis,
diversion colitis, Bechet's colitis, and indeterminate colitis.
[0370] FAEs are inhibitors of NF-.kappa.B activation and therefore
may be useful in treating inflammatory diseases such as Crohn's
disease and ulcerative colitis (Atreya et al., J Intern Med 2008,
263(6), 59106).
[0371] The efficacy of compounds of Formula (I) or Formula (II) for
treating inflammatory bowel disease can be evaluated using animal
models and in clinical trials. Useful animal models of inflammatory
bowel disease are known.
Irritable Bowel Syndrome
[0372] Irritable bowel syndrome is a disorder that affects the
large intestine and is typically characterized by abdominal pain or
cramping, a bloated feeling, flatulence, diarrhea or constipation
and/or mucus in the stool.
[0373] The efficacy of compounds of Formula (I) or Formula (II) for
treating irritable bowel syndrome can be evaluated using animal
models and in clinical trials. Useful animal models of inflammatory
bowel disease are known.
Asthma
[0374] Asthma is reversible airway obstruction in which the airway
occasionally constricts, becomes inflamed, and is lined with an
excessive amount of mucus. Symptoms of asthma include dyspnea,
wheezing, chest tightness, and cough. Asthma episodes may be
induced by airborne allergens, food allergies, medications, inhaled
irritants, physical exercise, respiratory infection, psychological
stress, hormonal changes, cold weather, or by other factors.
[0375] As an inhibitor of NF-.kappa.B activation and as shown in
animal studies (Joshi et al., US 2007/0027076) FAEs may be useful
in treating pulmonary diseases such as asthma and chronic
obstructive pulmonary disorder.
[0376] The efficacy of compounds of Formula (I) or Formula (II) for
treating asthma can be assessed using animal models and in clinical
trials.
Chronic Obstructive Pulmonary Disease
[0377] Chronic obstructive pulmonary disease (COPD), also known as
chronic obstructive airway disease, is a group of diseases
characterized by the pathological limitation of airflow in the
airway that is not fully reversible, and includes conditions such
as chronic bronchitis, emphysema, as well as other lung disorders
such as asbestosis, pneumoconiosis, and pulmonary neoplasms (see,
e.g., Barnes, Pharmacological Reviews 2004, 56(4), 515-548). The
airflow limitation is usually progressive and associated with an
abnormal inflammatory response of the lungs to noxious particles
and gases. COPD is characterized by a shortness of breath that
lasts for months or years, possibly accompanied by wheezing, and a
persistent cough with sputum production. COPD is most often caused
by tobacco smoking, although it can also be caused by other
airborne irritants such as coal dust, asbestos, urban pollution, or
solvents. COPD encompasses chronic obstructive bronchiolitis with
fibrosis and obstruction of small airways, and emphysema with
enlargement of airspaces and destruction of lung parenchyma, loss
of lung elasticity, and closure of small airways.
[0378] The efficacy of administering at least one compound of
Formula (I) or Formula (II) for treating chronic obstructive
pulmonary disease may be assessed using animal models of chronic
obstructive pulmonary disease and in clinical studies. For example,
murine models of chronic obstructive pulmonary disease are
known.
Neurodegenerative Disorders
[0379] Neurodegenerative diseases such as Parkinson's disease,
Alzheimer's disease, Huntington's disease and amyotrophic lateral
sclerosis are characterized by progressive dysfunction and neuronal
death. NF-.kappa.B inhibition has been proposed as a therapeutic
target for neurodegenerative diseases (Camandola and Mattson,
Expert Opin Ther Targets 2007, 11(2), 123-32).
Parkinson's Disease
[0380] Parkinson's disease is a slowly progressive degenerative
disorder of the nervous system characterized by tremor when muscles
are at rest (resting tremor), slowness of voluntary movements, and
increased muscle tone (rigidity). In Parkinson's disease, nerve
cells in the basal ganglia, e.g., substantia nigra, degenerate, and
thereby reduce the production of dopamine and the number of
connections between nerve cells in the basal ganglia. As a result,
the basal ganglia are unable to control smooth muscle movements and
coordinate changes in posture as normal, leading to tremor,
incoordination, and slowed, reduced movement (bradykinesia)
(Blandini, et al., Mol. Neurobiol. 1996, 12, 73-94).
[0381] The efficacy of compounds of Formula (I) or Formula (II) for
treating Parkinson's disease may be assessed using animal and human
models of Parkinson's disease and in clinical studies.
Alzheimer's Disease
[0382] Alzheimer's disease is a progressive loss of mental function
characterized by degeneration of brain tissue, including loss of
nerve cells and the development of senile plaques and
neurofibrillary tangles. In Alzheimer's disease, parts of the brain
degenerate, destroying nerve cells and reducing the responsiveness
of the maintaining neurons to neurotransmitters. Abnormalities in
brain tissue consist of senile or neuritic plaques, e.g., clumps of
dead nerve cells containing an abnormal, insoluble protein called
amyloid, and neurofibrillary tangles, twisted strands of insoluble
proteins in the nerve cell.
[0383] The efficacy of compounds of Formula (I) or Formula (II) for
treating Alzheimer's disease may be assessed using animal and human
models of Alzheimer's disease and in clinical studies.
Huntington's Disease
[0384] Huntington's disease is an autosomal dominant
neurodegenerative disorder in which specific cell death occurs in
the neostriatum and cortex (Martin, N Engl J Med 1999, 340,
1970-80). Onset usually occurs during the fourth or fifth decade of
life, with a mean survival at age of onset of 14 to 20 years.
Huntington's disease is universally fatal, and there is no
effective treatment. Symptoms include a characteristic movement
disorder (Huntington's chorea), cognitive dysfunction, and
psychiatric symptoms. The disease is caused by a mutation encoding
an abnormal expansion of CAG-encoded polyglutamine repeats in the
protein, huntingtin.
[0385] The efficacy of compounds of Formula (I) or Formula (II) for
treating Huntington's disease may be assessed using animal and
human models of Huntington's disease and in clinical studies.
Amyotrophic Lateral Sclerosis
[0386] Amyotrophic lateral sclerosis (ALS) is a progressive
neurodegenerative disorder characterized by the progressive and
specific loss of motor neurons in the brain, brain stem, and spinal
cord (Rowland and Schneider, N Engl J Med 2001, 344, 1688-1700).
ALS begins with weakness, often in the hands and less frequently in
the feet that generally progresses up an arm or leg. Over time,
weakness increases and spasticity develops characterized by muscle
twitching and tightening, followed by muscle spasms and possibly
tremors. The average age of onset is 55 years, and the average life
expectancy after the clinical onset is 4 years. The only recognized
treatment for ALS is riluzole, which can extend survival by only
about three months.
[0387] The efficacy compounds of Formula (I) or Formula (II) for
treating ALS may be assessed using animal and human models of ALS
and in clinical studies.
Others
[0388] Other diseases and conditions for which compounds of Formula
(I) or Formula (II) can be useful in treating include rheumatica,
granuloma annulare, lupus, autoimmune carditis, eczema,
sarcoidosis, and autoimmune diseases including acute disseminated
encephalomyelitis, Addison's disease, alopecia areata, ankylosing
spondylitis, antiphospholipid antibody syndrome, autoimmune
hemolytic anemia, autoimmune hepatitis, autoimmune inner ear
disease, bullous pemphigoid, Bechet's disease, celiac disease,
Chagas disease, chronic obstructive pulmonary disease, Crohn's
disease, dermatomyositis, diabetes mellitus type I, endometriosis,
Goodpasture's syndrome, Graves' disease, Guillain-Barre syndrome,
Hashimoto's disease, hidradenitis suppurativea, Kawasaki disease,
IgA neuropathy, idiopathic thrombocytopenic purpura, interstitial
cystitis, lupus erythematosus, mixed connective tissue disease,
morphea, multiple sclerosis, myasthenia gravis, narcolepsy,
neuromyotonia, pemphigus vulgaris, pernicious anemia, psoriasis,
psoriatic arthritis, polymyositis, primary biliary cirrhosis,
rheumatoid arthritis, schizophrenia, scleroderma, Sjogren's
syndrome, stiff person syndrome, temporal arteritis, ulcerative
colitis, vasculitis, vitiligo, acute disseminated
encephalomyelitis, myasthenia gravis, and Wegener's
granulomatosis.
Administration
[0389] Compounds of Formula (I) or Formula (II) and pharmaceutical
compositions thereof may be administered orally or by any other
appropriate route, for example, by infusion or bolus injection, by
absorption through epithelial or mucocutaneous linings (e.g., oral
mucosa, rectal, and intestinal mucosa, etc.). Other suitable routes
of administration include, for example, intradermal, intramuscular,
intraperitoneal, intravenous, subcutaneous, intranasal, epidural,
oral, sublingual, intracerebral, intravaginal, transdermal, rectal,
inhalation, or topical.
[0390] Administration may be systemic or local. Various delivery
systems are known, e.g., encapsulation in liposomes,
microparticles, microcapsules, capsules, etc., that may be used to
administer a compound and/or pharmaceutical composition.
[0391] The amount of a compound of Formula (I) or Formula (II) that
will be effective in the treatment of a disease in a patient will
depend, in part, on the nature of the condition and can be
determined by standard clinical techniques known in the art. In
addition, in vitro or in vivo assays may be employed to help
identify optimal dosage ranges. A therapeutically effective amount
of a compound of Formula (I) or Formula (II) to be administered may
also depend on, among other factors, the subject being treated, the
weight of the subject, the severity of the disease, the manner of
administration, and the judgment of the prescribing physician.
[0392] For systemic administration, a therapeutically effective
dose may be estimated initially from in vitro assays. For example,
a dose may be formulated in animal models to achieve a beneficial
circulating composition concentration range. Initial doses may also
be estimated from in vivo data, e.g., animal models, using
techniques that are known in the art. Such information may be used
to more accurately determine useful doses in humans. One having
ordinary skill in the art may optimize administration to humans
based on animal data.
[0393] A dose may be administered in a single dosage form or in
multiple dosage forms. When multiple dosage forms are used the
amount of compound contained within each dosage form may be the
same or different. The amount of a compound of Formula (I) or
Formula (II) contained in a dose may depend on the route of
administration and whether the disease in a patient is effectively
treated by acute, chronic, or a combination of acute and chronic
administration.
[0394] In certain embodiments an administered dose is less than a
toxic dose. Toxicity of the compositions described herein may be
determined by standard pharmaceutical procedures in cell cultures
or experimental animals, e.g., by determining the LD.sub.50 (the
dose lethal to 50% of the population) or the LD.sub.100 (the dose
lethal to 100% of the population). The dose ratio between toxic and
therapeutic effect is the therapeutic index. In certain
embodiments, a compound or metabolite thereof may exhibit a high
therapeutic index. The data obtained from these cell culture assays
and animal studies may be used in formulating a dosage range that
is not toxic for use in humans. A dose of a compound of Formula (I)
or Formula (II) may be within a range of circulating concentrations
in for example the blood, plasma, or central nervous system, that
include the effective dose and that exhibits little or no toxicity.
A dose may vary within this range depending upon the dosage form
employed and the route of administration utilized. In certain
embodiments, an escalating dose may be administered.
Combination Therapy
[0395] Methods provided by the present disclosure further comprise
administering one or more pharmaceutically active compounds in
addition to a morpholinoalkyl fumarate of Formula (I) or Formula
(II). Such compounds may be provided to treat the same disease or a
different disease than the disease being treated with the compound
of Formula (I) or Formula (II).
[0396] In certain embodiments, a compound of Formula (I) or Formula
(II) may be used in combination with at least one other therapeutic
agent. In certain embodiments, a compound of Formula (I) or Formula
(II) may be administered to a patient together with another
compound for treating diseases and conditions involving
immunological, autoimmune, and/or inflammatory processes including:
multiple sclerosis, psoriasis, irritable bowel disorder, ulcerative
colitis, arthritis, chronic obstructive pulmonary disease, asthma,
Parkinson's disease, Huntington's disease, amyotrophic lateral
sclerosis, and others. In certain embodiments, a compound of
Formula (I) or Formula (II) may be administered to a patient
together with another compound for treating multiple sclerosis,
psoriasis, irritable bowel disorder, ulcerative colitis, arthritis,
chronic obstructive pulmonary disease, asthma, Parkinson's disease,
Huntington's disease, or amyotrophic lateral sclerosis.
[0397] A compound of Formula (I) or Formula (II) and the at least
one other therapeutic agent may act additively or, in certain
embodiments, synergistically. The at least one additional
therapeutic agent may be included in the same dosage form as a
compound of Formula (I) or Formula (II) or may be provided in a
separate dosage form. Methods provided by the present disclosure
can further include, in addition to administering a compound of
Formula (I) or Formula (II), administering one or more therapeutic
agents effective for treating the same or different disease than
the disease being treated by a compound of Formula (I) or Formula
(II). Methods provided by the present disclosure include
administration of a compound of Formula (I) or Formula (II) and one
or more other therapeutic agents provided that the combined
administration does not inhibit the therapeutic efficacy of the
compound of Formula (I) or Formula (II), or any pharmacologically
active metabolite thereof, and/or does not typically produce
significant and/or substantial adverse combination effects.
[0398] In certain embodiments, dosage forms comprising a compound
of Formula (I) or Formula (II) may be administered concurrently
with the administration of another therapeutic agent, which may be
part of the same dosage form as, or in a different dosage form
than, that comprising a compound of Formula (I) or Formula (II). A
compound of Formula (I) or Formula (II) may be administered prior
to, or subsequent to, administration of another therapeutic agent.
In certain embodiments, the combination therapy may comprise
alternating between administering a compound of Formula (I) or
Formula (II) and administering another therapeutic agent, e.g., to
minimize adverse drug effects associated with a particular drug.
When a compound of Formula (I) or Formula (II) is administered
concurrently with another therapeutic agent that potentially may
produce an adverse drug effect including, for example, toxicity,
the other therapeutic agent may be administered at a dose that
falls below the threshold at which the adverse drug effect is
elicited.
[0399] In certain embodiments, dosage forms comprising a compound
of Formula (I) or Formula (II) may be administered with one or more
substances to enhance, modulate and/or control release,
bioavailability, therapeutic efficacy, therapeutic potency,
stability, and the like of a compound of Formula (I) or Formula
(II). For example, to enhance the therapeutic efficacy of a
compound of Formula (I) or Formula (II), the compound of Formula
(I) or Formula (II) may be co-administered with, or a dosage form
comprising a compound of Formula (I) or Formula (II) may comprise,
one or more active agents to increase the absorption or diffusion
of a compound of Formula (I) or Formula (II) from the
gastrointestinal tract to the systemic circulation, or to inhibit
degradation of the compound of Formula (I) or Formula (II) in the
blood of a patient. In certain embodiments, a compound of Formula
(I) or Formula (II) may be co-administered with an active agent
having pharmacological effects that enhance the therapeutic
efficacy of a compound of Formula (I) or Formula (II).
[0400] In certain embodiments, a compound of Formula (I), a
compound of Formula (II), or a pharmaceutical composition thereof
may be administered to a patient for treating psoriasis in
combination with a therapy or another therapeutic agent known or
believed to be effective in treating psoriasis. Drugs useful for
treating psoriasis include, for example, steroids such as
flurandrenolide, fluocinonide, alclometasone, amcinonide, desonide,
halcinonide, triamcinolone, clobetasol, clocortolone, mometasone,
desoximetasone, and halobetasol; anti-rheumatics such as
etanercept, infiximab, and adalimumab; immunosuppressive agents
such as cyclosporine, alefacept, and efalizumab; psoralens such as
methoxsalen; and other such as calcipotriene, methotrexate,
hydrocortisone/pramoxine, acitretin, betamethasone/calcipotriene,
tazaraotene, benzocaine/pyrilamine/zinc oxide, and ustekinumab.
[0401] In certain embodiments, a compound of Formula (I), a
compound of Formula (II), or a pharmaceutical composition thereof
may be administered to a patient for treating inflammatory
arthritis such as rheumatoid arthritis, juvenile rheumatoid
arthritis, psoriatic arthritis, and ankylosing spondylitis in
combination with a therapy or another therapeutic agent known or
believed to be effective in treating inflammatory arthritis such as
rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic
arthritis, and ankylosing spondylitis.
[0402] Drugs useful for treating rheumatoid arthritis include, for
example, non-steroidal anti-inflammatory agents such as ibuprofen,
ketoprofen, salicylate, diclofenac, nabumetone, naproxen,
meloxicam, sulindac, flurbiprofen, indomethacin, tolmetin,
piroxicam, fenoprofen, oxaprozin, and etodolac; antiheumatics such
as entanercept, adalimumab, infliximab, hydroxychloroquine,
leflunomide, azathioprine, penicillamine, methotrexate, anakinra,
auranofin, rituximab, aurothioglucose, tocilizumab, and golimumab;
COX-2 inhibitors such as celecoxib and vadecoxib; corticosteroids
such as triamcinolone; glucocorticoids such as methylprednisolone
and prednisone; and others such as sulfasalazine.
[0403] Drugs useful for treating juvenile rheumatoid arthritis
include, for example, adalimumab, abatacept, and infliximab.
[0404] Drugs useful for treating psoriatic arthritis include, for
example, etanercept, adalimumab, triamcinolone, cortisone,
infliximab, and golimumab.
[0405] Drugs useful for treating ankylosing spondylitis include,
for example, adalimumab, celecoxib, diclofenac, etanercept,
golimumab, indomethacin infliximab, naptoxen, olsalazine,
salicylates, sulfindac, and triamcinolone.
[0406] In certain embodiments, a compound of Formula (I), a
compound of Formula (II), or a pharmaceutical composition thereof
may be administered to a patient for treating psoriatic arthritis
in combination with a therapy or another therapeutic agent known or
believed to be effective in treating psioriatic arthritis. Drugs
useful for treating psioriatic arthritis include, for example,
etanercept, adalimumab, triamcinolone, cortisone, infliximab, and
golimumab.
[0407] In certain embodiments, a compound of Formula (I), a
compound of Formula (II), or a pharmaceutical composition thereof
may be administered to a patient for treating autoimmune diseases
such as lupus in combination with a therapy or another therapeutic
agent known or believed to be effective in treating autoimmune
diseases such as lupus. Drugs useful for treating lupus include,
for example, hydroxychloroquine, triamcinolone, salicylate,
azathioprine, and abetimus.
[0408] In certain embodiments, a compound of Formula (I), a
compound of Formula (II), or a pharmaceutical composition thereof
may be administered to a patient for treating multiple sclerosis in
combination with a therapy or another therapeutic agent known or
believed to be effective in treating multiple sclerosis. Drugs
useful for treating multiple sclerosis include, for example,
interferon .beta.-1a, interferon .beta.-1b, glatiramer, modafinil,
azathioprine, prednisolone, mycophenolate mofetil, mitoxantrone,
and natalizumab. Other examples of drugs useful for treating MS
include, for example, corticosteroids such as methylprednisolone;
IFN-.beta. such as IFN-.beta.1a and IFN-.beta.1b; glatiramer
acetate; monoclonal antibodies that bind to the very late antigen-4
(VLA-4) integrin such as natalizumab; immunomodulatory agents such
as FTY 720 sphinogosie-1 phosphate modulator and COX-2 inhibitors
such as BW755c, piroxicam, and phenidone; and neuroprotective
treatments including inhibitors of glutamate excitotoxicity and
iNOS, free-radical scavengers, and cationic channel blockers;
memantine; AMPA antagonists such as topiramate; and glycine-site
NMDA antagonists.
[0409] In certain embodiments, a compound of Formula (I), a
compound of Formula (II), or a pharmaceutical composition thereof
may be administered to a patient for treating inflammatory bowel
disease in combination with a therapy or another therapeutic agent
known or believed to be effective in treating inflammatory bowel
disease. Drugs useful for treating inflammatory bowel disease
include, for example, cromolyn and mercaptopurine; and more
particularly for treating Crohn's disease include certolizumab,
budesonide, azathioprine, sulfasalazine, metronidazole, adalimumab,
mercaptopurine, infliximab, mesalamine, and natalizumab; and for
treating ulcerative colitis include balsalazide, infliximab,
azathioprine, mesalamine, and cyclosporine.
[0410] In certain embodiments, a compound of Formula (I), a
compound of Formula (II), or a pharmaceutical composition thereof
may be administered to a patient for treating irritable bowel
syndrome in combination with a therapy or another therapeutic agent
known or believed to be effective in treating irritable bowel
syndrome. Drugs useful for treating irritable bowel syndrome
include, for example, lactobacillus acidophilus, dicylmine,
atropine, hyoscyamine, phenobarbital, scopolamine, venlafaxine,
chloridazepoxide, clidinium, alosetron, psyllium, cholestyramine,
rifaximin, and tegaserod.
[0411] In certain embodiments, compounds of Formula (I) or Formula
(II) and pharmaceutical compositions thereof may be administered to
a patient for treating asthma in combination with a therapy or
another therapeutic agent known or believed to be effective in
treating asthma, or in certain embodiments, a disease, disorder, or
condition associated with asthma. Examples of drugs useful in
treating asthma include, for example, albuterol, aminophylline,
beclomethasone, bitolterol, budesonide, cromolyn, ephedrine,
epinephrine, flunisolide, fluticasone, formoterol, hydrocortisone,
isoproterenol, levalbuterol, methylprednisolone, prednisolone,
prednisone, pirbuterol, metaproterenol, racepinephrine, omalizumab,
oxytriphylline, mometusone, montelukast, nedocromil, oxtriphylline,
pirbuterol, salmeterol, terbutaline, theophylline, triamcinolone,
zafirlukast, and zileuton.
[0412] In certain embodiments, compounds of Formula (I) or Formula
(II) and pharmaceutical compositions thereof may be administered to
a patient for treating chronic obstructive pulmonary disease in
combination with a therapy or another therapeutic agent known or
believed to be effective in treating chronic obstructive pulmonary
disease, or in certain embodiments, a disease, disorder, or
condition associated with chronic obstructive pulmonary disease.
Examples of drugs useful for treating chronic obstructive pulmonary
disease include, for example, albuterol, arformoterol,
azithromycin, bitolterol, epinephrine, fluticasone, formoterol,
ipratropium, isoproterenol, levabuterol, metaproterenol,
pirbuterol, racepinephrine, salmeterol, and tiotropium. Useful
drugs for treating chronic obstructive pulmonary disease further
include, for example, bronchodialators such as .beta.2 agonists
such as salbutamol, bambuterol, clenbuterol, fenoterol, and
formoterol; M3 antimuscarinics such as ipratropium; leukotriene
antagonists such as montelukast, pranlukast, and zafirlukast;
cromones such as cromoglicate and nedocromil; xanthines such as
theophylline; corticosteroids such as beclomethasone, mometasone,
and fluticasone; and TNF antagonists such as infliximab,
adalimumab, and etanercept. Other treatments for chronic
obstructive pulmonary disease include oxygen therapy, and pulmonary
rehabilitation.
[0413] In certain embodiments, compounds of Formula (I) or Formula
(II) and pharmaceutical compositions thereof may be administered to
a patient for treating angiogenesis in combination with a therapy
or another therapeutic agent known or believed to be effective in
treating angiogenesis. Useful drugs for treating angiogenesis
include, for example, angiostatin, endostatin, vitaxin,
bevacizumab, thalidomide, batimastat, marimastat,
carboxyamidotraizole, TNP-470, CM101, IFN-.alpha., IL-12, platelet
factor-4, suramin, SU5416, thrombospondin, VEGFR, angiostatic
steroids, cartilage-derived angiogenesis inhibitory factor, matrix
metalloproteinase inhibitors, 2-methoxyestradiol, tecogalan,
thrombospondin, prolactin, .alpha..sub.v.beta..sub.3 inhibitors,
and linomide.
[0414] In certain embodiments, compounds of Formula (I) or Formula
(II) and pharmaceutical compositions thereof may be administered to
a patient for treating transplant rejection in combination with a
therapy or another therapeutic agent known or believed to be
effective in treating transplant rejection. Useful drugs for
treating transplant rejection include, for example, calcineurin
inhibitors such as cyclosporine and tacrolimus, mTOR inhibitors
such as sirolimus and everolimus, anti-proliferatives such as
azathioprine and mycophenolic acid; monoclonal anti-IL2R.alpha.
receptor antibodies including basiliximab and daclizumab; and
polyclonal anti-T-cell antibodies including anti-thymocyte globulin
and anti-lymphocyte globulin.
[0415] In certain embodiments, compounds of Formula (I) or Formula
(II) and pharmaceutical compositions thereof may be administered to
a patient for treating transplantation rejection in combination
with a therapy or another therapeutic agent known or believed to be
effective in treating transplantation rejection. Examples of drugs
useful in transplantation rejection include, for example,
corticosteroids such as dexamethasone, prednisolone, and
prednisone; globulins such as antilymphocyte globulin and
antithymocyte globulin; macrolide immunosuppressants such as
sirolimus, tacrolimus, and everolimus; mitotic inhibitors such as
azathiprine, cylophosphamide, and methotrexate; monoclonal
antibodies such as basiliximab, daclizumab, infliximab, muromonoab;
fungal metabolites such as cyclosporine; and others such as
glatiramer and mycophenolate.
[0416] In certain embodiments, compounds of Formula (I) or Formula
(II) and pharmaceutical compositions thereof may be administered to
a patient for treating cardiac insufficiency in combination with a
therapy or another therapeutic agent known or believed to be
effective in treating cardiac insufficiency. Useful drugs for
treating cardiac insufficiency include, for example,
antitensin-modulating agents, diuretics such as furosemide,
bumetanie, hydrochlorothiazide, chlorthalidone, chlorthiazide,
spironolactone, eplerenone: beta blockers such as bisoprolol,
carvedilol, and metroprolol; positive inotropes such as digoxin,
milrinone, and dobutamine; alternative vasodilators such as
isosorbide dinitrate/hydralazine; aldosterone receptor antagonists;
recombinant neuroendocrine hormones such as nesiritide; and
vasopressin receptor antagonists such as tolvaptan and
conivaptan.
[0417] In certain embodiments, compounds of Formula (I) or Formula
(II) and pharmaceutical compositions thereof may be administered to
a patient for treating a mitochondrial disease such as a
neurodegenerative disease in combination with a therapy or another
therapeutic agent known or believed to be effective in treating a
mitochondrial disease such as a neurodegenerative disease. In
certain embodiments, a neurodegenerative disease is chosen from
Alzheimer's disease, Parkinson's disease, Huntington's disease, and
amyotrophic lateral sclerosis.
[0418] Therapeutic agents useful for treating Parkinson's disease
include, for example, dopamine precursors such levodopa, dopamine
agonists such as bromocriptine, pergolide, pramipexole, and
ropinirole, MAO-B inhibitors such as selegiline, anticholinergic
drugs such as benztropine, trihexyphenidyl, tricyclic
antidepressants such as amitriptyline, amoxapine, clomipramine,
desipramine, doxepin, imipramine, maprotiline, nortriptyline,
protriptyline, amantadine, and trimipramine, some antihistamines
such as diphenhydramine; antiviral drugs such as amantadine; and
beta blockers such as propranolol.
[0419] Useful drugs for treating Alzheimer's disease include, for
example, roloxifene, vitamin E, donepezil, tacrine, rivastigmine,
galantamine, and memantine.
[0420] Useful drugs for treating symptoms of Huntington's disease
include, for example, antipsychotics such as haloperidol,
chlorpromazine and olanzapine to control hallucinations, delusions
and violent outbursts; antidepressants such as fluoxetine,
sertraline, and nortriptyline to control depression and
obsessive-compulsive behavior; tranquilizers such as
benzodiazepines, paroxetine, venflaxin and beta-blockers to control
anxiety and chorea; mood stabilizers such as lithium, valproate,
and carbamzepine to control mania and bipolar disorder; and
botulinum toxin to control dystonia and jaw clenching. Useful drugs
for treating symptoms of Huntington's disease further include
selective serotonin reuptake inhibitors (SSRIs) such as fluoxetine,
paroxetine, sertraline, escitalopram, citalopram, fluvosamine;
norepinephrine; serotonin reuptake inhibitors (NSRI) such as
venlafaxine and duloxetine; benzodiazepines such as clonazepam,
alprazolam, diazepam, and lorazepam; tricyclic antidepressants such
as amitriptyline, nortriptyline, and imipramine; atypical
antidepressants such as busipirone, bupriopion, and mirtazepine for
treating the symptoms of anxiety and depression; atomoxetine,
dextroamphetamine, and modafinil for treating apathy symptoms;
amantadine, memantine, and tetrabenazine for treating chorea
symptoms; citalopram, atomoxetine, memantine, rivastigmine, and
donepezil for treating cognitive symptoms; lorazepam and trazedone
for treating insomnia; valproate, carbamazepine and lamotrigine for
treating symptoms of irritability; SSRI antidepressants such as
fluoxetine, paroxetine, sertaline, and fluvoxamine; NSRI
antidpressants such as venlafaxine; others such as mirtazepine,
clomipramine, lomotrigine, gabapentin, valproate, carbamazepine,
olanzapine, rispiridone, and quetiapine for treating symptoms of
obsessive-compulsive disorder; haloperidol, quetiapine, clozapine,
risperidone, olanzapine, ziprasidone, and aripiprazole for treating
psychosis; and pramipexole, levodopa and amantadine for treating
rigidity.
[0421] Useful drugs for treating ALS include, for example,
riluzole. Other drugs of potential use in treating ALS include, for
example, memantine, tamoxifen, thalidomide, ceftriaxone, sodium
phenyl butyrate, celecoxib, glatiramer acetate, busipirone,
creatine, minocycline, coenzyme Q10, oxandrolone, IGF-1,
topiramate, xaliproden, and indinavir. Drugs such as baclofen and
diazepam can be useful in treating spasticity associated with
ALS.
[0422] In certain embodiments, a compound of Formula (I), a
compound of Formula (II), or a pharmaceutical composition thereof
may be administered to a patient in combination with a therapy or
another therapeutic agent known or believed to be effective in
inhibiting TNF function.
[0423] Examples of drugs known to inhibit TNF function include, for
example, infliximab, adalimumab, etanercept, certolizumab,
goliimumab, pentoxifylline, quanylhydrozone, thalidomide,
flavonoids such as narigenin, resveratol and quecetin, alkaloids
such as lycorine, terpenes such as acanthoic acid, fatty acids such
as 13-HOA, and retinoids such as retinoic acid.
EXAMPLES
[0424] The following examples describe in detail the synthesis of
morpholinoalkyl fumarates of Formula (I) or Formula (II),
properties of compounds of Formula (I) or Formula (II), and uses of
compounds of Formula (I) or Formula (II). It will be apparent to
those skilled in the art that many modifications, both to materials
and methods, may be practiced without departing from the scope of
the disclosure.
General Experimental Protocols
[0425] All reagents and solvents that are purchased from commercial
suppliers are used without further purification or manipulation
procedures.
[0426] Proton NMR (400 MHz) and carbon NMR spectra (125 MHz) are
recorded on a Varian AS 400 NMR spectrometer equipped with an
autosampler and data processing software. CDCl.sub.3 (99.8% D),
DMSO-d.sup.6 (99.9% D), or MeOH-d.sup.4 (99.8+% D), and
acetonitrile-d.sup.3 are used as solvents unless otherwise noted.
The CHCl.sub.3, DMSO-d.sup.5, or MeOH-d.sup.3 solvent signals are
used for calibration of the individual spectra. Analytical thin
layer chromatography (TLC) is performed using a Whatman, Schleicher
& Schuell TLC and MK6F silica gel plates (2.5.times.7.5 cm, 250
.mu.m layer thickness). Melting points are recorded in glass
capillaries using a Stanford Research Systems (SRS) Optimelt
Automated Melting Point System, S/N 78047. Analytical LC/MS is
performed on a Waters 2790 separation module equipped with a Waters
Micromass QZ mass spectrometer, a Waters 996 photodiode detector,
and a Merck Chromolith UM2072-027 or Phenomenex Luna C-18
analytical column. Mass-guided preparative HPLC purification of
final compounds is performed using an instrument equipped with a
Waters 600 controller, ZMD Micromass spectrometer, a Waters 2996
photodiode array detector, and a Waters 2700 Sample Manager.
Acetonitrile/water gradients containing 0.05% formic acid are used
as eluents in both analytical and preparative HPLC experiments.
Compound isolation from aqueous solvent mixtures, e.g.,
acetonitrile/water/0.05% formic acid, is accomplished by primary
lyophilization (freeze drying) of the frozen solutions under
reduced pressure at room temperature using manifold freeze dryers
such as a Heto Drywinner DW 6-85-1, a Heto FD4, or a VIRTIS
Freezemobile 25 ES equipped with high vacuum pumps. When the
isolated compound has ionizable functional groups such as an amino
group or a carboxylic acid, lyophilization is performed in the
presence of a slight excess of one molar (1 M) hydrochloric acid to
yield the purified compounds as the corresponding hydrochloride
salts (HCl-salts) or the corresponding protonated free carboxylic
acids. When the isolated compound has ionizable functional groups
such as a carboxylic acid, lyophilization is performed in the
presence of equimolar amounts of sodium hydrogen carbonate
(NaHCO.sub.3) to yield the purified compounds as the corresponding
sodium salts (Na-salts). Optionally, the isolated materials are
further purified by flush silica gel column chromatography,
optionally employing Biotage pre-packed silica gel cartridges.
Suitable organic solvents such as ethyl acetate (EtOAc), hexane
(Hxn), n-heptane (Hptn), or mixtures and/or gradients thereof are
used as eluents to yield the target compounds as colorless, viscous
oils or solids after evaporation of the solvents. Chemical names
are generated with the Chemistry 4-D Draw Pro Version 7.01c (Draw
Chemical Structures Intelligently.COPYRGT. 1993-2002) from
ChemInnovation Software, Inc., San Diego, USA).
[0427] Non-commercially available starting materials are
synthesized from commercially available starting materials, and by
adapting methods well known in the art.
General Procedure A
Nucleophilic Substitution of 1-Haloalkylmorpholine Derivatives with
Monoalkyl Fumarate
[0428] (2E)-3-(Alkoxycarbonyl)prop-2-enoic acid (alkyl hydrogen
fumarate), (2E)-3-(tert-butoxycarbonyl)prop-2-enoic acid
(tert-butyl hydrogen fumarate), or fumaric acid (FA) (1.0
equivalents) is dissolved in 5-10 mL/3.0 mmol of an inert solvent
such as N-methylpyrrolidone (NMP), N,N-dimethylformamide,
N,N-dimethylacetamide (DMA, DMAc), acetonitrile (MeCN),
dimethylsulfoxide (DMSO), tetrahydrofuran (THF), toluene, or
mixtures thereof. To the solution, 0.8 to 1.2 equivalents of an
appropriate inorganic base such as cesium hydrogen carbonate
(CsHCO.sub.3), cesium carbonate (Cs.sub.2CO.sub.3), or potassium
carbonate (K.sub.2CO.sub.3) is added. Alternatively, 0.8 to 1.2
equivalents of a silver salt such silver(I) oxide (Ag.sub.2O) or
silver(I) carbonate (Ag.sub.2CO.sub.3); an organic secondary or
tertiary base such as dicyclohexylamine (DCHA), triethylamine
(TEA), diisopropylethylamine (DIEA), tetrabutylammonium hydroxide
(TBAOH), amidine; or a guanidine-based base such as
1,5-diazabicyclo[4.3.0]non-5-ene (DBN),
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), or
1,1,3,3-tetramethylguanidine (TMG), can be employed. The
corresponding alkali, silver, di-, tri- and tetraalkylammonium,
amidine, or guanide salt of monoalkyl fumarate can also be
pre-formed. The solution is stirred for 10-60 min at room
temperature followed by addition of 0.8 to 1.2 equivalents of an
appropriately functionalized 1-halo alkylmorpholine. The reaction
mixture is stirred overnight at a temperature between 40.degree. C.
to 100.degree. C. After cooling to room temperature, insolubles can
optionally be filtered off and the reaction mixture diluted with
water and an appropriate organic solvent such as methyl tert-butyl
ether (MTBE), diethyl ether (Et.sub.2O), ethylacetate (EtOAc), or
mixtures thereof. After phase separation, the aqueous phase is
extracted several times with the same solvent. The combined organic
extracts are washed with water, brine, and dried over anhydrous
magnesium sulfate (MgSO.sub.4). After filtration, the organic
solvents are removed under reduced pressure using a rotary
evaporator. If required, the crude reaction products are further
purified by well-known purification techniques such as silica gel
flash column chromatography (i.e., Biotage), mass-guided
reversed-phase preparative HPLC/lyophilization, precipitation, or
crystallization.
General Procedure B1
Activation of Carboxylic Acid Derivatives with Dehydration Agents
for Aminolysis or Alcoholysis
[0429] A monoalkyl fumarate (1.0 equivalents) is reacted at
temperature from ca. 0.degree. C. (ice bath) to room temperature
with 1.0-1.5 equivalents of a carbodiimide dehydration agent such
as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAC, EDC),
N,N'-diisopropylcarbodiimide (DIC), N,N'-dicyclohexylcarbodiimide
(DCC) in an inert solvent such as dichloromethane (DCM),
N,N-dimethylformamide, N-methylpyrrolidone (NMP), or
N,N-dimethylacetamide (DMA, DMAc) (ca. 3 mL/mmol). 1.0-1.5
equivalents of 1-hydroxyalkyl morpholine dissolved in the same
solvent and, optionally, in the presence of a catalytic or
stoichiometric amount of 4-(N,N-dimethylaminopyridine) (DMAP) is
added at a temperature from ca. 0.degree. C. to room temperature.
When the amine is a salt form, an equimolar amount of an organic
tertiary base, such as triethylamine (TEA), or
diisopropylethylamine (DIEA) may be added to free the amine base
prior to the coupling step. The reaction mixture is stirred for 4
to 12 hours at room temperature. Optionally, the organic solvents
are removed under reduced pressure using a rotary evaporator and
the residue diluted with an appropriate extraction solvent such as
diethyl ether (Et.sub.2O), methyl tert-butyl ether (MTBE), ethyl
acetate (EtOAc), or others. The procedures described in Procedure A
for product isolation and purification may be employed.
General Procedure B2
Activation of Carboxylic Acid Derivatives with Chlorination Agents
and Aminolysis
[0430] A monoalkyl fumarate (1.0 equivalents) is reacted with
oxalyl chloride (1.0-1.5 equivalents) in anhydrous dichloromethane
(DCM), ca. 3 mL/mmol, at a temperature of ca. 0.degree. C. (ice
bath) in the presence of a catalytic amount of
N,N-dimethylformamide for 1 to 3 hours. The solvents are removed
under reduced pressure using a rotary evaporator and the crude
material is dissolved in anhydrous dichloromethane (DCM), ca. 3
mL/mmol. 1.0 to 1.5 equivalents of a 1-hydroxyalkyl morpholine in
anhydrous dichloromethane (DCM), ca. 3 mL/mmol, is added drop-wise
at ca. 0.degree. C. (ice bath), optionally in the presence of a
catalytic amount of 4-(N,N-dimethylamino)pyridine (DMAP). When the
1-hydroxyalkyl morpholine is a salt form, an equimolar amount of a
base such as triethylamine (TEA), diisopropylethylamine (DIEA), or
others, is added prior to the coupling step. The reaction is
stirred overnight with warming to room temperature, the solvents
optionally removed under reduced pressure using a rotary
evaporator, and then diluted with an appropriate extraction solvent
such as diethyl ether (Et.sub.2O), methyl tert-butyl ether (MTBE),
ethyl acetate (EtOAc), or others. The procedures described in
Procedure A for product isolation and purification may be
employed.
General Procedure C
Preparation of Mono-Esters of Fumaric Acid
[0431] Coupling Reaction of morpholin-4-ylalkyl-1-ol with Fumaric
Acid
##STR00046##
[0432] Fumaric acid (1.0 eq.) is dissolved in an inert solvent such
as dichloromethane (DCM), N,N-dimethylformamide (DMF),
N-methylpyrrolidone (NMP), or N,N-dimethylacetamide (DMA, DMAc)
(ca. 3 mL/mmol) and the solution is treacted with 1.0-1.5 eq. of a
carbodiimide dehydration agent such as
1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDAC, EDC),
N,N-diisopropylcarbodiimide (DIC), N,N-dicyclohexyl-carbodiimide
(DCC) at a temperature from ca. 0.degree. C. (ice bath) to room
temperature. The mixture is then reacted with a solution of an
appropriately functionalized of morpholin-4-ylalkyl-1-ol (1.0-1.5
eq.) in the same solvent. Optionally, a catalytic or stoichiometric
amount of 4-(N,N-dimethylaminopyridine (DMAP) is added to the
mixture at a temperature from ca. 0.degree. C. to room temperature.
When the amine is in a salt form, an equimolar amount of an organic
tertiary base, such as triethylamine (TEA), or
diisopropylethylamine (DIEA) may be added to free the amine base
prior to the coupling step. The reaction mixture is stirred for 4
to 12 hours at room temperature. Optionally the organic solvents
are removed under reduced pressure using a rotary evaporator and
the residue diluted with an appropriate extraction solvent such as
diethyl ether (Et.sub.2O), methyl tert-butyl ether (MTBE), ethyl
acetate (EtOAc), or others, Water is added to the reaction mixture,
the aqueous phase was acidified using 1N hydrochloric acid until
aqueous pH reaches to pH .about.2. After phase separation, the
aqueous phase is extracted several times with the same solvent. The
combined organic extracts are washed with water, brine, and dried
over anhydrous magnesium sulfate (MgSO.sub.4). After filtration,
the organic solvents are removed under reduced pressure using a
rotary evaporator, if required, the crude reaction products are
further purified by well-known purification techniques such as
silica gel flash column chromatography (i.e., Biotage), mass-guided
reversed-phase preparative HPLC/lyophilization, precipitation, or
crystallization to yield the pure desired product.
General Procedure D
Preparation of Bis-esters
Coupling Reaction of morpholin-4-ylalkyl-1-ol with Fumaric Acid
##STR00047##
[0434] Fumaric acid (1.0 eq.) is dissolved in an inert solvent such
as dichloromethane (DCM), N,N-dimethylformamide (DMF),
N-methylpyrrolidone (NMP), or: N,N-dimethylacetamide (DMA, DMAc)
(ca. 3 mL/mmol) and the solution is reacted with 2.0-2.5 eq. of a
carbodiimide dehydration agent such as
1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDAC, EDC),
N,N-diisopropylcarbodiimide (DIC), N,N-dicyclohexyl-carbodiimide
(DCC) at a temperature from ca. 0.degree. C. (ice bath) to room
temperature. A solution of an appropriately functionalized of
morpholin-4-ylalkyl-1-ol (2.0-2.5 eq.) in the same solvent and,
optionally, a catalytic or stoichiometric amount of
4-(N,N-dimethylaminopyridine (DMAP) are added to the above mixture
at a temperature from ca. 0.degree. C. to room temperature, When
the amine is in a salt form, an equimolar amount of an organic
tertiary base, such as triethylamine (TEA), or
diisopropylethylamine (DIEA) may be added to free up the amine base
prior to the coupling step. The reaction mixture is then stirred
for 4 to 12 hours at room temperature or until the reaction goes to
completion (TLC or HPLC). The organic solvents are removed under
reduced pressure using a rotary evaporator and the residue is
diluted with an appropriate extraction solvent such as diethyl
ether (Et.sub.2O), methyl tert-butyl ether (MTBE), ethyl acetate
(EtOAc), or others. After phase separation, the aqueous phase is
extracted several times with the same solvent, The combined organic
extracts are washed with water, brine, and dried over anhydrous
magnesium sulfate (MgSO.sub.4), After filtration, the organic
solvents are removed under reduced pressure using a rotary
evaporator. If required, the crude reaction product is further
purified by well-known purification techniques such as silica gel
flash column chromatography (i.e., Biotage), mass-guided
reversed-phase preparative HPLC/lyophilization, precipitation, or
crystallization to give the pure bis ester.
Example 1
Methyl (2-morpholinoethyl)fumarate (Methyl 2-morpholin-4-ylethyl
(2E)but-2-ene-1,4-dioate) (1)
##STR00048##
[0436] Following general Procedure A, methyl hydrogen fumarate
(MHF) (26 g, 0.2 mol) was activated with
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC)
(47.75 g, 0.25 mol) in 200 mL of dichloromethane (DCM) at ca.
0.degree. C. 2-Morpholin-4-ylethyl-1-ol (26.2 g, 0.2 mol) and
4-N,N-dimethylaminopyridine (DMAP) (1 g, 0.008 mol) were added to
the activated carboxylic acid. After synthesis, 38 g (81%, yield)
of the title compound was isolated as a viscous-oil.
[0437] .sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 6.88 (m, 2H),
4.35-4.33 (m, 2H), 3.82 (s, 3H), 3.72-3.70 (m, 4H), 2.699-2.68 (m,
2H), 2.67-2.51 (m, 4H), MS (ESI): m/z 245.11 (M+H).sup.+.
Example 2
Methyl (2-morpholinoethyl)fumarate HCl salt (Methyl
2-morpholin-4-ylethyl (2E)but-2-ene-1,4-dioate HCl salt) (2)
##STR00049##
[0439] Following general Procedure B, Methyl 2-morpholin-4-ylethyl
(2E)but-2-ene-1,4-dioate (38 g, 0.156 mol) was dissolved in methyl
tert-butyl ether (200 mL). The resulting clear reaction mixture is
cooled to 0.degree. C. (ice bath). 1.1 equivalent of hydrochloride
in dioxane (4M) was slowly added over a period of 30 minutes.
During this period the product starts to precipitate/crystallize
out as off-white solid. The solid product was separated by
filtration and the filter-cake was washed with methyl tert-butyl
ether (100 mL). The filter-cake was dried under vacuum oven at
40.degree. C. to 38 g (82.5% yield) of the title compound as a
white solid.
[0440] .sup.1H NMR (MeOH-D.sub.3, 400 MHz): .delta. 6.93 (m, 2H),
4.61-4.58 (m, 2H), 3.95-3.89 (broad m, 4H), 3.81 (s, 3H), 3.57-3.55
(m, 2H), 3.31-3.29 (m, 4H), MS (ESI): m/z 245.11 (M+H).sup.+.
Melting point: 214.3.degree. C.
Example 3
Methyl (3-morpholinopropyl)fumarate (Methyl 3-morpholin-4-ylpropyl
(2E)but-2-ene-1,4-dioate) (3)
##STR00050##
[0442] Following general Procedure A, methyl hydrogen fumarate
(MHF) (8.9 g, 0.068 mol) was activated with
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC)
(15.28 g, 0.08 mol) in 200 mL of dichloromethane (DCM) at ea.
0.degree. C. 3-Morpholin-4ylpropyl-1-ol (10 g, 0.068 mol) and
4-N,N-dimethylaminopyridine (DMAP) (500 mg, 0.004 mol) were added
to the activated carboxylic acid. After synthesis, 9 g (50.8%
yield) of the title compound was afforded as a viscous-oil.
[0443] .sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 6.87-6.86 (m,
2H), 4.28-4.26 (m, 2H), 3.82 (s, 3H), 3.72-3.70 (m, 4H), 2.44 (m,
4H), 1.89-1.86 (m, 2H), 1.63 (m, 2H), MS (ESI): m/z 259.13
(M+H).sup.+.
Example 4
Methyl (3-morpholinopropyl)fumarate HCl salt (Methyl
3-morpholin-4ylpropyl (2E)but-2-ene-1,4-dioate HCl salt) (4)
##STR00051##
[0445] Following general Procedure B, Methyl 3-morpholin-4-ylpropyl
(2E)but-2-ene-1,4-dioate (9 g, 0.035 mol) was dissolved in methyl
tert-butyl ether (50 mL). The resulting clear reaction mixture is
cooled to 0.degree. C. (ice bath). 1.2 equivalent of hydrochloride
in dioxane (4M) was slowly added over a period of 30 minutes.
During this period the product starts to precipitate/crystallize
out as off-white solid. The solid product was separated by
filtration and the filter-cake was washed with methyl tert-butyl
ether (50 mL). The filter-cake was dried under vacuum oven at
40.degree. C. to 10 g (98.0% yield) of the title compound as a
white solid.
[0446] .sup.1H NMR (MeOH-D.sub.3, 400 MHz): .delta. 6.88 (m, 2H),
4.34-4.31 (m, 2H), 3.95-3.89 (broad m, 4H), 3.82 (s, 3H), 3.30-3.27
(m, 6H), 3.31-3.29 (m, 2H), MS (ESI): m/z 259.13 (M+H).sup.+.
Melting point: 185.2.degree. C.
Example 5
Methyl (4-morpholinobutyl)fumarate (Methyl 4-morpholin-4ylbutyl
(2E)but-2-ene-1,4-dioate) (5)
##STR00052##
[0448] Following general procedure A, methyl hydrogen fumarate
(MHF) (26 g, 0.2 mol) was activated with
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC)
(47.75 g, 0.25 mol) in 200 mL, of dichloromethane (DCM) at ca.
0.degree. C. 4-Morpholin-4ylbutyl-1-ol (31.8 g, 0.2 mol) and
4-N,N-dimethylaminopyridine (DMAP) (1 g, 0.008 mol) were added to
the activated carboxylic acid. After work-up and isolation, 45 g
(83.3% yield) of the title compound was afforded as a
viscous-oil.
[0449] .sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 6.86-6.86 (m,
2H), 4.24-4.21 (m, 2H), 3.81 (s, 3H), 3.71-3.69 (m, 4H), 2.43-2.34
(m, 6H), 1.74-1.69 (m, 2H), 1.60-1.56 (m, 2H), MS (ESI): m/z 272.14
(M+H).sup.+.
Example 6
Methyl (4-morpholinobutyl)fumarate HCl salt (Methyl
4-morpholin-4ylbutyl (2E)but-2-ene-1,4-dioate HCl salt) (6)
##STR00053##
[0451] Following general procedure B, Methyl 4-morpholin-4ylbutyl
(2E)but-2-ene-1,4-dioate (45 g, 0.166 mol) was dissolved in methyl
tert-butyl ether (200 mL). The resulting clear reaction mixture is
cooled to 0.degree. C. (ice bath). 1.0 equivalent of hydrochloride
in dioxane (4M) was slowly added over a period of 30 minutes.
During this period the product starts to precipitate/crystallize
out as off-white solid. The solid product was separated by
filtration and the filter-cake was washed with methyl tert-butyl
ether (200 mL). The filter-cake was dried under vacuum oven at
40.degree. C. to 38 g (65.0% yield) of the title compound as a
white solid.
[0452] .sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 6.90-6.82 (m,
2H), 4.34-4.24 (m, 4H), 4.01-3.97 (m, 2H), 3.82 (s, 3H), 3.48-3.45
(m, 2H), 3.06-3.03 (m, 2H), 3.00-2.84 (m, 2H), 2.09-1.81 (m, 2H),
1.79-1.77 (m, 2H) MS (ESI): m/z 272.14 (M+H).sup.+. Melting point:
145.5.degree. C.
Example 7
Methyl (5-morpholinopentyl)fumarate (Methyl 5-morpholin-4ylpentyl
(2E)but-2-ene-1,4-dioate) (7)
##STR00054##
[0454] Following general procedure A, methyl hydrogen fumarate
(MHF) (13.0 g, 0.1 mol) was activated with
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC)
(22.9 g, 0.12 mol) in 50 mL of dichloromethane (DCM) at ca.
0.degree. C. 5-Morpholin-4ylpentyl-1-01 (17.3 g, 0.1 mol) and
4-N,N-dimethylaminopyridine (DMAP) (100 mg) were added to the
activated carboxylic acid. After work-up and isolation, 11 g (28.5%
yield) of the title compound was afforded as a viscous-oil.
[0455] .sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 6.86 (m, 2H),
4.24-4.21 (m, 2H), 3.81 (s, 3H), 3.71-3.69 (m, 4H), 2.43-2.34 (m,
6H), 1.74-1.69 (m, 2H), 1.60.1.56 (m, 4H), MS (ESI): m/z 287.16
(M+H).sup.+.
Example 8
Methyl (5-morpholinopentyl)fumarate HCl salt (Methyl
5-morpholin-4ylpentyl (2E)but-2-ene-1,4-dioate HCl salt) (8)
##STR00055##
[0457] Following general procedure B, Methyl 5-morpholin-4ylpentyl
(2E)but-2-ene-1,4-dioate (11 g, 0.0385 mol) was dissolved in methyl
tert-butyl ether (100 mL). The resulting clear reaction mixture is
cooled to 0.degree. C. (ice bath). 1.2 equivalent of hydrochloride
in dioxane (4M) was slowly added over a period of 30 minutes.
During this period the product starts to precipitate/crystallize
out as off-white solid. The solid product was separated by
filtration and the filter-cake was washed with methyl tert-butyl
ether (100 mL). The filter-cake was dried under vacuum oven at
40.degree. C. to 110 g (81.3% yield) of the title compound as a
white solid.
[0458] .sup.1H NMR (MeOH-D.sub.3, 400 MHz): .delta. 6.82-6.81 (m,
2H), 4.26-4.22 (m, 2H), 4.08-4.03 (broad m, 2H), 3.79 (s, 3H),
3.50-3.31 (broad m, 2H), 3.30-3.29 (m, 2H), 3.18-3.14 (m, 4H),
1.85-1.74 (m, 4H), 1.52-1.45 (m. 2H), MS (ESI): m/z 287.16.13
(M+H).sup.+. Melting point: 150.0.degree. C.
Example 9
Methyl (6-morpholinohexyl)fumarate (Methyl
6-morpholin-4-ylhexyl(2E)but-2-ene-1,4-dioate) (9)
##STR00056##
[0460] Following general procedure A, methyl hydrogen fumarate
(MHF) (2.0 g, 15.3 mmol) was activated with
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC)
(182 g, 20.0 mmol) in 25 mL of dichloromethane (DCM) at ca.
0.degree. C. 6-Morpholin-4ylhexyl-1-ol (3 g, 16.0 mmol) and
4-N,N-dimethylaminopyridine (DMAP) (100 mg) were added to the
activated carboxylic acid. After work-up and isolation, 3 g (62.6%
yield) of the title compound was afforded as a viscous-oil.
[0461] .sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 6.86 (m, 2H),
4.24-4.21 (m, 2H), 3.81 (s, 3H), 3.71-3.69 (m, 4H), 2.43-2.34 (m,
6H), 1.74-1.69 (m, 4H), 1.60.1.56 (m, 4H), MS (ESI): m/z 301.18
(M+H).sup.+.
Example 10
Methyl (6-morpholinohexyl)fumarate HCl salt (Methyl
6-morpholin-4ylhexyl(2E)but-2-ene-1,4-dioate HCl salt) (10)
##STR00057##
[0463] Following general procedure B, Methyl
6-morpholin-4ylhexyl(2E)but-2-ene-1,4-dioate (3 g, 10 mmol) was
dissolved in methyl tert-butyl ether (25 mL). The resulting clear
reaction mixture is cooled to 0.degree. C. (ice bath). 1.2
equivalent of Hydrochloride in dioxane (4M) was slowly added over a
period of 30 minutes. During this period the product starts to
precipitate/crystallize out as off-white solid. The solid product
was separated by filtration and the filter-cake was washed with
methyl tert-butyl ether (25 mL). The filter-cake was dried under
vacuum oven at 40.degree. C. to 3 g (93.75% yield) of the title
compound as a white solid.
[0464] .sup.1H NMR (MeOH-D.sub.3, 400 MHz): .delta. 6.82-6.81 (m,
2H), 4.24-4.20 (m, 2H), 4.23-4.08 (broad m, 2H), 3.79 (s, 3H),
3.55-3.45 (broad m, 2H), 3.31-3.29 (m, 2H), 3.17-3.12 (m, 4H),
1.77-1.72 (m, 4H), 1.47-1.45 (m. 4H), MS (ESI): m/z 301.18
(M+H).sup.+. Melting point: 104.1.degree. C.
Example 11
Ethyl (2-morpholinoethyl)fumarate (11)
##STR00058##
[0466] Following the procedure of Example 1, and replacing methyl
hydrogen fumarate with ethyl hydrogen fumarate provides the title
compound (II). The reaction of the free base with HCl in dioxane
and following the procedure of Example 2 affords the corresponding
HCl salt.
Example 12
Propyl (2-morpholinoethyl)fumarate (13)
##STR00059##
[0468] Following the procedure of Example 1, and replacing methyl
hydrogen fumarate with propyl hydrogen fumarate provides the title
compound (13). The reaction of the free base with HCl in dioxane
and following the procedure of Example 2 affords the corresponding
HCl salt.
Example 13
Butyl (2-morpholinoethyl)fumarate (15)
##STR00060##
[0470] Following the procedure of Example 1, and replacing methyl
hydrogen fumarate with propyl hydrogen fumarate provides the title
compound (15). The reaction of the free base with HCl in dioxane
and following the procedure of Example 2 affords the corresponding
HCl salt.
Example 14
Pentyl (2-morpholinoethyl)fumarate (17)
##STR00061##
[0472] Following the procedure of Example 1, and replacing methyl
hydrogen fumarate with pentyl hydrogen fumarate provides the title
compound (17). The reaction of the free base with HCl in dioxane
and following the procedure of Example 2 affords the corresponding
HCl salt.
Example 15
(4-Morpholinobutyl)fumarate (39)
##STR00062##
[0474] Following general procedure C, t-butyl hydrogen fumarate
(MHF) (0.2 mot) is activated with
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC)
(47.75 g, 0.25 mol) in 200 mL of dichloromethane (DCM) at ca.
0.degree. C. 4-Morpholin-4ylbutyl-1-ol (31.8 g, 0.2 mol) and
4-N,N-dimethylaminopyridine (DMAP) (1 g, 0.008 mot) were added to
the activated carboxylic acid. After work-up and isolation, the
crude material is reacted with 50% trifluoroacetic acid in DCM. The
free acid is purified by mass-guided preparative HPLC to afford the
title compound (40).
Example 16
(5-Morpholinopentyl)fumarate (40)
##STR00063##
[0476] Following the procedure of Example 15, and replacing
4-morpholin-4ylbutyl-1-ol with 5-morpholin-4ylpentyl-1-ol provides
the title compound (40).
Example 17
Methyl (6-morpholinohexyl)fumarate (Methyl
6-morpholin-4-ylhexyl(2E)but-2-ene-1,4-dioate) (9)
##STR00064##
[0478] Following the procedure of Example 15, and replacing
4-morpholin-4ylbutyl-1-ol with 6-morpholin-4ylhexyl-1-ol provides
the title compound (41).
Example 18
Bis(3-Morpholinopropyl)fumarate (42)
##STR00065##
[0480] Following general procedure D, fumaric acid (0.2 mol) is
activated with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
hydrochloride (EDAC) (0.25 mol) in 200 mL of dichloromethane (DCM)
at ca, 0.degree. C. 3-Morpholin-4ylpropyl-1-ol (0.2 mol) and
4-N,N-dimethylaminopyridine (DMAP) (1 g, 0.008 mol) were added to
the activated carboxylic acid. After work-up and isolation, the
crude is purified by mass-guided preparative HPLC to afford the
title compound (42).
Example 19
(4-Morpholinobutyl)fumarate (43)
##STR00066##
[0482] Following the procedure of Example 18, and replacing
3-morpholin-4ylpropyl-1-ol with 5-morpholin-4ylbutyl-1-ol provides
the title compound (43).
Example 20
(5-Morpholinopentyl)fumarate (44)
##STR00067##
[0484] Following the procedure of Example 18, and replacing
3-morpholin-4ylpropyl-1-ol with 5-morpholin-4ylpentyl-1-ol provides
the title compound (44).
Example 21
(5-Morpholinopentyl)fumarate (45)
##STR00068##
[0486] Following the procedure of Example 18, and replacing
3-morpholin-4ylpropyl-1-ol with 6-morpholin-4ylhexyl-1-ol provides
the title compound (45).
[0487] The synthetic and biological examples described in this
application are offered to illustrate the compounds, pharmaceutical
compositions and methods provided herein and are not to be
construed in any way as limiting their scope. In the examples, all
temperatures are in degrees Celsius (unless otherwise indicated).
Compounds that can be prepared in accordance with the methods
provided herein along with their biological and PK data are
presented in following Tables. The syntheses of these
representative compounds are carried out in accordance with the
methods set forth above.
Exemplary Compounds Provided Herein
[0488] The following compounds have been or can be prepared
according to the synthetic methods described herein.
TABLE-US-00001 TABLE 1 Exemplary Compounds of the Disclosure
Compound ID Structure 1 ##STR00069## 2 ##STR00070## 3 ##STR00071##
4 ##STR00072## 5 ##STR00073## 6 ##STR00074## 7 ##STR00075## 8
##STR00076## 9 ##STR00077## 10 ##STR00078## 11 ##STR00079## 12
##STR00080## 13 ##STR00081## 14 ##STR00082## 15 ##STR00083## 16
##STR00084## 17 ##STR00085## 18 ##STR00086## 19 ##STR00087## 20
##STR00088## 21 ##STR00089## 22 ##STR00090## 23 ##STR00091## 24
##STR00092## 25 ##STR00093## 26 ##STR00094## 27 ##STR00095## 28
##STR00096## 29 ##STR00097## 30 ##STR00098## 31 ##STR00099## 32
##STR00100## 33 ##STR00101## 34 ##STR00102## 35 ##STR00103## 36
##STR00104## 37 ##STR00105## 38 ##STR00106## 39 ##STR00107## 40
##STR00108## 41 ##STR00109## 42 ##STR00110## 43 ##STR00111## 44
##STR00112## 45 ##STR00113##
Description 1
Methods for Determining Stability of Morpholinoalkyl Fumarates In
Vitro
[0489] Certain morpholinoalkyl fumarates of the disclosure may or
may not themselves be pharmacologically active, and are metabolized
in vivo to produce a pharmacologically active metabolite. For a
prodrug, it can be desirable that the prodrug remains intact (i.e.,
uncleaved) while in the systemic circulation and be cleaved (i.e.,
to release the parent drug) in the target tissue. Alternatively, it
can be desirable that the prodrug remains intact (i.e., uncleaved)
while in the gastrointestinal tract and be cleaved (i.e., to
release the parent drug) after being absorbed or taken up from the
gastrointestinal lumen, e.g., in either the enterocytes lining the
gastrointestinal lumen or in the blood. For pharmacologically
active compounds, it can be desirable that the compound remains
intact in the gastrointestinal tract.
[0490] A useful level of stability can at least in part be
determined by the mechanism and pharmacokinetics of the prodrug or
active compound. In general, prodrugs or active compounds that are
more stable in pancreatin or colonic wash assay, and are more
labile in rat plasma, human plasma, rat liver S9, and/or human
liver S9 preparations, can be useful as an orally administered
prodrug or active compound. In general, prodrugs or active
compounds that are more stable in rat plasma, human plasma, rat
liver S9, and/or human liver S9 preparations, and which are more
labile in cell homogenate preparations such CaCo2 S9 preparations,
can be useful as systemically administered prodrugs or active
compounds and/or can be more effective in delivering a prodrug or
active compound to a target tissue. In general, prodrugs or active
compounds that are more stable in a range of pH physiological
buffers (pH 6.0 to pH 8.5) can be more useful as orally
administered prodrugs or active compounds. In general, prodrugs or
active compounds that are more labile in cell homogenate
preparations, such CaCo2 S9 preparations, can be intracellularly
metabolized. The results of tests, such as those described in this
example, for determining the enzymatic or chemical cleavage of
compounds in vitro can be used to select prodrugs for in vivo
testing.
[0491] The stabilities of prodrugs or active compounds can be
evaluated in one or more in vitro systems using a variety of
preparations following methods known in the art. For example,
methods used to determine the stability of prodrugs in Caco2 S9
homogenate, rat liver S9, rat plasma, porcine pancreatin, rat
colonic wash, and pH 8.0 buffer are described herein.
[0492] CaCo2 S9 homogenate is prepared using the following
procedure. CaCo2 cells are grown in culture for 21 days prior to
harvesting. Culture medium is removed from the culture vessel and
the monolayer is rinsed twice with 10-15 mL chilled phosphate
buffered saline (PBS) buffer. PBS buffer (7-10 mL) is added to the
flask and the cells scraped from the growth surface and transferred
to a centrifuge tube. The cells are pelleted by centrifugation at
1,500 rpm for 5 min at 4.degree. C. The supernatant is removed and
the cell pellet washed with ice cold PBS and re-pelleted by
centrifugation. The supernatant is removed and the pellet
re-suspended in cell lysis buffer (0.15M KCl and 10 mM sodium
phosphate buffer, pH 7.4). Cells are lysed by sonication at
4.degree. C. using a probe sonicator. The lysed cells are then
transferred to vials and centrifuged at 1,600 rpm for 10 min at
4.degree. C. to remove intact cells, nuclei, and large cellular
debris. The supernatant is removed and transferred to a tube for
centrifugation at 8,600 rpm for 20 min at 4.degree. C. After
centrifugation, the resulting supernatant, representing the CaCo2
cell homogenate S9 fraction, is carefully removed and aliquoted
into vials for storage at -80.degree. C. until the time of use. At
the time of use, CaCo2 S9 lysate is diluted to 0.5 mg/mL in 0.1M
Tris buffer, pH 7.4.
[0493] Rat liver S9 (XenoTech, Lenexa, Kans.; R1000.59, 20 mg/mL)
is diluted to 0.5 mg/mL in 0.1 M potassium phosphate buffer at pH
7.4 and 1 mM NADPH cofactor.
[0494] Rat plasma (Pel-Freez.RTM. Biologicals, Rogers, Ark.; 36150)
is used as obtained from the supplier.
[0495] Porcine pancreatin (Sigma Aldrich, St. Louis, Mo.;
P1625-100G) is diluted to 10 mg/mL in 0.1M Tris buffer, pH 7.4.
[0496] To prepare the rat colonic wash, the colon between the
ceacum and rectum is resected from a euthanized rat. Five to 10 mL
of PBS pH 7.4 buffer (depending on the weight of the rat) is
flushed into the lumen of the large intestine and collected into a
250 mL glass beaker at 0.degree. C. (ice bath). The colonic wash is
transferred into 10 mL conical tubes using a 10 mL syringe fitted
with a filter. Samples of 0.5 mL colonic wash are stored at
-80.degree. C. until the time of use. Colonic wash is used without
dilution.
[0497] The enzymatic stability assays for a compound in CaCo2 S9,
rat liver S9, rat plasma, pig pancreatin, and rat colonic wash are
performed using the following procedure. Ninety (90) .mu.L of
lysate is aliquoted to designated tubes on a cluster plate. The
lysate is pre-incubated for 10 min at 37.degree. C. With the
exception of the t(0) time point, 10 .mu.L of a 400 .mu.M solution
of test compound in 0.1M Tris buffer, pH 7.4 is added to multiple
wells, representing different incubation times. The samples are
incubated at 37.degree. C. At each time point, the reaction is
quenched by adding 300 .mu.L of 100% ethanol. The samples are
thoroughly mixed, the tubes transferred to a V-bottom plate, and
stored at -20.degree. C. For the t(0) time point, the lysate is
quenched with 300 .mu.L of ice cold 100% ethanol, thoroughly mixed,
10 .mu.L of 400 .mu.M test compound is added and mixed, and the
sample tube transferred to a V-bottom plate and stored at
-20.degree. C. For analysis, 180 .mu.L from each sample is
transferred to a 96 well V-bottom plate and sealed. After all time
points are collected, the plate is centrifuged for 10 min at 5600
rpm at 4.degree. C. One-hundred fifty (150) .mu.L from each well is
then transferred to a 96 well round bottom plate. Samples are
analyzed using LC/MS/MS to determine the concentrations of the
compound and/or metabolite thereof.
[0498] For the pH 8.0 stability studies, 190 .mu.L of 150 mM
NaH.sub.2PO.sub.4 buffer pH 8.0 is added to each sample tube. Ten
(10) .mu.L of 20 mM test compound is added to each tube and mixed.
The samples are incubated for 60 min at 37.degree. C. Following
incubation, the samples are transferred to room temperature and 800
.mu.L of 50% acetonitrile in water is added to each tube. Samples
are analyzed using LC/MS/MS to determine the concentrations of the
compound and/or metabolite thereof.
[0499] LC/MS/MS analysis for MHF is performed using an API 4000
equipped with an Agilent 1100 HPLC and a Leap Technologies
autosampler. An HPLC Phenomenex Onyx Monolithic C18 (CH0-7644)
column at a temperature of 35.degree. C., flow rate of 2.0 mL/min,
injection volume of 30 .mu.L, and a 3-min run time is used. The
mobile phase AI is 0.1% formic acid in water and Mobile phase AII
is 0.1% formic acid in acetonitrile. The gradient is 98% AI/2% AII
at time 0; 98% AI/2% AII at time 0.1 min; 5% AI/95% AII at time 1.4
min; 5% AI/95% AII at time 2.2 min; 98% AI/2% AII at time 2.3 min;
and 98% AI/2% AII at time 3.0 min. MHF content is determined using
negative ion mode (Q1 128.94; Q2 71).
Description 2
Methyl Hydrogen Fumarate Bioavailability Following Oral
Administration of Morpholinoalkyl Fumarates
[0500] Rats are obtained commercially and are pre-cannulated in the
jugular vein. Animals are conscious at the time of the experiment.
All animals are fasted overnight and until 4 hours post-dosing of a
compound of the disclosure.
[0501] Blood samples (0.3 mL/sample) are collected from all animals
prior to dosing and at different time-points up to 24 h post-dose
into tubes containing EDTA. Two aliquots (100 .mu.L each) are
quenched with 300 .mu.L methanol and stored at -20.degree. C. prior
to analysis.
[0502] To prepare analysis standards, 90 .mu.L of rat blood is
quenched with 300 .mu.L methanol followed by 10 .mu.L of spiking
standard and/or 20 .mu.L of internal standard. The sample tubes are
vortexed for at least 2 min and then centrifuged at 3,400 rpm for
20 min. The supernatant is then transferred to an injection vial or
plate for analysis by LC-MS-MS.
[0503] To prepare samples for analysis, 20 .mu.L of internal
standard is added to each quenched sample tube. The sample tubes
are vortexed for at least 2 min and then centrifuged at 3,400 rpm
for 20 min. The supernatant is then transferred to an injection
vial or plate for analysis by LC/MS/MS.
[0504] LC/MS/MS analysis can be performed using an API 4000 (MS12)
equipped with Agilent 1100 HPLC and a Leap Technologies
autosampler. The following HPLC column conditions are used: HPLC
column: Onyx Monolithic C18 Phenomex (PN CHO-7644), 35.degree. C.;
flow rate 2.0 mL/min; injection volume 30 .mu.L; run time 3 min;
mobile phase A: 0.1% formic acid in water; mobile phase B: 0.1%
formic acid in acetonitrile (ACN); gradient: 98% A/2% B at 0.0 min;
98% A/2% B at 0.1 min; 5% A/95% B at 1.4 min; 5% A/95% B at 2.2
min; 98% A/2% B at 2.3 min; and 98% A/2% B at 3.0 min. MHF is
monitored in negative ion mode.
[0505] Non-compartmental analysis is performed using WinNonlin
software (v.3.1 Professional Version, Pharsight Corporation,
Mountain View, Calif.) on individual animal profiles. Summary
statistics on major parameter estimates is performed for C.sub.max
(peak observed concentration following dosing), T.sub.max (time to
maximum concentration is the time at which the peak concentration
is observed), AUC.sub.(0-t) (area under the plasma
concentration-time curve from time zero to last collection time,
estimated using the log-linear trapezoidal method),
AUC.sub.(0-.infin.), (area under the plasma concentration time
curve from time zero to infinity, estimated using the log-linear
trapezoidal method to the last collection time with extrapolation
to infinity), and t.sub.1/2,z (terminal half-life).
[0506] A compound of the disclosure is administered by oral gavage
to groups of four to six adult male Sprague-Dawley rats (about 250
g). Animals are conscious at the time of the experiment. A compound
of the disclosure is orally or colonically administered in 3.4%
Phosal at a dose of 70 mg-equivalents MHF per kg body weight.
[0507] The percent relative bioavailability (F %) of the
administered compound or metabolite thereof is determined by
comparing the area under the respective concentration vs time curve
(AUC) following oral or colonic administration of a compound of the
disclosure with the AUC of the concentration vs time curve
following intravenous administration of the compound of the
disclosure, respectively, on a dose-normalized basis.
[0508] The % F can be reported as the mean % F of all animals dosed
orally with the compound of the disclosure at the specified
level.
[0509] The oral bioavailability (% F) values of the compounds
tested along with the comparative compound, DMF, in rats and in
monkeys are set forth in Table 2, below.
TABLE-US-00002 TABLE 2 Oral Bioavailability of Exemplary Compounds
Oral Oral Compound Bioavailability Bioavailability ID Structure
(Rat) F (%) (Monkey) F (%) 2 ##STR00114## 25.7 NA 4 ##STR00115##
17.2 NS 6 ##STR00116## 41.3 78.2 8 ##STR00117## 27.5 NA 10
##STR00118## 10.5 NA DMF ##STR00119## 37 61
Description 3
Animal Model for Assessing Therapeutic Efficacy of Morpholinoalkyl
Fumarates for Treating Multiple Sclerosis
Animals and Experimental Autoimmune Encephalomyelitis Induction
[0510] Female C57BL/6 mice, 8-10 weeks old (Harlan Laboratories,
Livermore, Calif.), are immunized subcutaneously in the flanks and
mid-scapular region with 200 .mu.g of myelin oligodendrocyte
glycoprotein peptide 35-55 (MOG.sub.35-55) (synthesized by
Invitrogen) emulsified (1:1 volume ratio) with complete Freund's
adjuvant (CFA) (containing 4 mg/mL Mycobacterium tuberculosis).
Emulsion is prepared by the syringe-extrusion method with two glass
Luer-Lock syringes connected by a 3-way stopcock. Mice are also
given an intraperitoneal injection of 200 ng pertussis toxin (List
Biological Laboratories, Inc, Campbell, Calif.) on the day of
immunization and on day two post immunization. Mice are weighed and
examined daily for clinical signs of experimental autoimmune
encephalomyelitis (EAE). Food and water is provided ad libitum and
once animals start to show disease, food is provided on the cage
bottom. All experiments are approved by the Institutional Animal
Care and Use Committee.
Clinical Evaluation
[0511] Mice are scored daily beginning on day 7 post immunization.
The clinical scoring scale is as follows (Miller and Karplus,
Current Protocols in Immunology 2007, 15.1.1-15.1.18): 0=normal;
1=limp tail or hind limb weakness (defined by foot slips between
bars of cage top while walking); 2=limp tail and hind limb
weakness; 3=partial hind limb paralysis (defined as no weight
bearing on hind limbs but can still move one or both hind limbs to
some extent); 4=complete hind limb paralysis; 5=moribund state
(includes forelimb paralysis) or death.
Treatment
[0512] Compound(s) of the disclosure are dissolved in 0.5%
methocellulose/0.1% Tween80 in distilled water and administered by
oral gavage twice daily starting from day 3 post-immunization until
termination. Dexamethasone is dissolved in 1.times.PBS buffer and
administered subcutaneously once daily. Treatment groups are, for
example, as follows: vehicle alone, 15 mg/kg DMF, 20 mg/kg compound
of the disclosure, and 1 mg/kg dexamethasone.
Alternate Animal Models of Multiple Sclerosis
[0513] The following experiment confirmed that MHF is the active
moiety of both MHF prodrugs DMF and the compounds of the disclosure
and examined the relationship between MHF exposure and effect in
animal models of multiple sclerosis (MS). Efficacy of
representative compound of the disclosure and DMF is compared in
the MOG35-55 mouse EAE model of multiple sclerosis. C57BL/6 mice (6
females) are injected subcutaneously with MOG35-55 peptide in CFA
with Mycobacterium tuberculosis. Pertussis toxin (200 mg) is
injected IV on Day 0 and Day 2 post-immunization. Animals received
oral test compound or DMF (90 mg-eq MHF/kg twice daily) or vehicle
on Days 3 to 29. Daily EAE clinical disease scores (5 point scale)
are recorded. End of study MHF blood levels are determined by
LC/MS/MS.
Description 4
Use of an Animal Model to Assess Efficacy in Treating Psoriasis
[0514] The severe, combined immunodeficient (SCID) mouse model can
be used to evaluate the efficacy of compounds for treating
psoriasis in humans (Boehncke, Ernst Schering Res Found Workshop
2005, 50, 213-34; and Bhagavathula et al., J Pharmacol Expt'l
Therapeutics 2008, 324(3), 938-947).
[0515] SCID mice are used as tissue recipients. One biopsy for each
normal or psoriatic volunteer (human) is transplanted onto the
dorsal surface of a recipient mouse. Treatment is initiated 1 to 2
weeks after transplantation. Animals with the human skin
transplants are divided into treatment groups. Animals are treated
twice daily for 14 days. At the end of treatment, animals are
photographed and then euthanized. The transplanted human tissue
along with the surrounding mouse skin is surgically removed and
fixed in 10% formalin and samples obtained for microscopy.
Epidermal thickness is measured. Tissue sections are stained with
an antibody to the proliferation-associated antigen Ki-67 and with
an anti-human CD3.sup.+ monoclonal antibody to detect human T
lymphocytes in the transplanted tissue. Sections are also probed
with antibodies to c-myc and .beta.-catenin. A positive response to
treatment is reflected by a reduction in the average epiderma
thickness of the psoriatic skin transplants. A positive response is
also associated with reduced expression of Ki-67 in
keratinocytes.
Alternate Animal Models of Multiple Sclerosis and Psoriasis
[0516] Imidquimod model of skin inflammation (Fits et al The
Journal of Immunology, 2009, 182: 5836-5845). 10-12 week old
BALB/c, Il17c+/+ or Il17c-/-, or Il17re+/+ or Il17re-/- mice were
administered 50 mg Aldara cream (5% Imidquimod in Graceway, 3M) in
the shaved back and right ear daily for 5 days. Clinical scoring
and ear thickness measurements were performed daily. Scoring was
based upon the manifestation of psoriatic symptoms, such as
erythema, scaling and thickness: 0, No disease. 1, Very mild
erythema with very mild thickening and scaling involving a small
area. 2, Mild erythema with mild thickening and scaling involving a
small area. 3, Moderate erythema with moderate thickening and
scaling (irregular and patchy) involving a small area (<25%). 4,
Severe erythema with marked thickening and scaling (irregular and
patchy) involving a moderate area (25-50%). 5, Severe erythema with
marked thickening and scaling (irregular and patchy) involving a
large area (>50%). Ear and back tissue were harvested on day 5
for histological evaluation.
[0517] Efficacy of compounds of the disclosure and DMF is compared
in the imiquimod (IMQ) mouse model of psoriasis. Balb/c mice (10
males/group) received daily topical IMQ (5% cream) on shaved back
and right ear for 5 days as described above. Animals received oral
dose of a representative compound of the disclosure or DMF (45 or
90 mg-eq MMF/kg twice daily) or vehicle from Day -5 to Day 5.
Erythema score is the primary outcome measure.
[0518] The Erythema score values of the compounds tested at an oral
dose of 90 mg-eq MMF/kg BID for 10 days in male Balb/C mice are set
forth in Table 3, below. The data shows that the compounds of the
disclosure are equipotent to DMF.
TABLE-US-00003 TABLE 3 Efficacy of Exemplary Compounds in Psoriasis
Model Erythema Score (% Relative to Compound ID IMQ)* 2 69 4 73.3 6
57.1 8 59.3 10 66.7 DMF 74.1 *oral dose of 90 mg-eq MMF/kg BID for
10 days in male Balb/C mice
Description 5
Animal Model for Assessing Therapeutic Efficacy of Morpholinoalkyl
Fumarates for Treating Multiple Sclerosis
[0519] Experiments are conducted on female mice aged 4-6 weeks
belong to the C57BL/6 strain weighing 17-20 g. Experimental
autoimmune encephalomyelitis (EAE) is actively induced using 95%
pure synthetic myelin oligodendrocyte glycoprotein peptide 35-55
(MOG.sub.35-55) (synthesized by Invitrogen). Each mouse is
anesthetized and receives 200 .mu.g of MOG.sub.35-55 peptide and 15
.mu.g of Saponin extract from Quilija bark emulsified in 100 .mu.L
of phosphate-buffered saline. A 25 .mu.L volume is injected
subcutaneously over four flank areas. Mice are also
intraperitoneally injected with 200 ng of pertussis toxin in 200
.mu.l of PBS. A second, identical injection of pertussis toxin is
given after 48 h.
[0520] A compound of the disclosure is administered at varying
doses. Control animals receive 25 .mu.l of DMSO. Daily treatment
extends from day 26 to day 36 post-immunization. Clinical scores
are obtained daily from day 0 post-immunization until day 60.
Clinical signs are scored using the following protocol: 0, no
detectable signs; 0.5, distal tail limpness, hunched appearance and
quiet demeanor; 1, completely limp tail; 1.5, limp tail and
hindlimb weakness (unsteady gait and poor grip with hind limbs); 2,
unilateral partial hind limb paralysis; 2.5, bilateral hind limb
paralysis; 3, complete bilateral hindlimb paralysis; 3.5, complete
hindlimb paralysis and unilateral forelimb paralysis; 4, total
paralysis of hind limbs and forelimbs (Eugster et al., Eur J
Immunol 2001, 31, 2302-2312).
[0521] Inflammation and demyelination are assessed by histology on
sections from the CNS of EAE mice. Mice are sacrificed after 30 or
60 days and whole spinal cords are removed and placed in 0.32 M
sucrose solution at 4.degree. C. overnight. Tissues are prepared
and sectioned. Luxol fast blue stain is used to observe areas of
demyelination. Haematoxylin and eosin staining is used to highlight
areas of inflammation by darkly staining the nuclei of mononuclear
cells. Immune cells stained with H&E are counted in a blinded
manner under a light microscope. Sections are separated into gray
and white matter and each sector is counted manually before being
combined to give a total for the section. T cells are immunolabeled
with anti-CD3+ monoclonal antibody. After washing, sections are
incubated with goat anti-rat HRP secondary antibody. Sections are
then washed and counterstained with methyl green. Splenocytes
isolated from mice at 30 and 60 days post-immunization are treated
with lysis buffer to remove red blood cells. Cells are then
re-suspended in PBS and counted. Cells at a density of about
3.times.10.sup.6 cells/mL are incubated overnight with 20 .mu.g/mL
of MOG peptide. Supernatants from stimulated cells are assayed for
IFN-.gamma. protein levels using an appropriate mouse IFN-.gamma.
immunoassay system.
Description 6
Use of an Animal Model to Assess Efficacy in Treating Inflammatory
Bowel Disease
[0522] Animal models of inflammatory bowel disease are described by
Jurjus et al., J Pharmaocol Toxicol Methods 2004, 50, 81-92;
Villegas et al., Int'l Immunopharmacol 2003, 3, 1731-1741; and
Murakami et al., Biochemical Pharmacol 2003, 66, 1253-1261. For
example, the following protocol can be used to assess the efficacy
of a compound of the disclosure for treating inflammatory bowel
disease.
[0523] Female ICR mice are used. Mice are divided into treatment
groups. Groups are given either water (control), 5% DSS in tap
water is given at the beginning of the experiment to induce
colitis, or various concentrations of test compound. After
administering test compound for 1 week, 5% DSS in tap water is also
administered to the groups receiving test compound for 1 week. At
the end of the experiment, all mice are sacrificed and the large
intestine is removed. Colonic mucosa samples are obtained and
homogenized. Proinflammatory mediators (e.g., IL-1.alpha.,
IL-1.beta., TNF-.alpha., PGE2, and PGF2.alpha.) and protein
concentrations are quantified. Each excised large intestine is
histologically examined and the damage to the colon scored.
Description 7
Clinical Trial for Assessing Efficacy in Treating Asthma
[0524] Adult subjects (nonsmokers) with stable mild-to-moderate
asthma are enrolled (see, e.g., Van Schoor and Pauwels, Eur Respir
J 2002, 19, 997-1002). A randomized, double-blind,
placebo-controlled, two-period crossover design is used. On
screening day 1, patients undergo a methacholine challenge (<8
mg/mL). The baseline forced expiratory volume in one second (FEV1)
prior to each subsequent challenge must be within 15% of the
screening baseline FEV1 obtained at the first visit. A neurokinin
challenge (1.times.10.sup.-6 mol/mL) on screening day 2 is
performed 24-72 h later. Study-period one commences within 10 days
after visit two. First, a methacholine and a neurokinin-A (NKA)
challenge is performed on days 1 and 0, respectively. At visit
four, test compound is administered at an appropriate dose and for
an appropriate period of time. On the last 2 days of the treatment
period, methacholine and NKA challenges are repeated. Following
treatment-period one, there is a washout period of about 5 weeks,
following which the patients crossed over to another medication or
placebo in study period two, which is identical to period one.
Pulmonary function tests are performed using a spirometer. The
metacholine challenge is performed by inhaling doubling
concentrations of methacholine until the FEV1 falls by >20% of
the post-diluent baseline FEV1 of that day as described by
Cockcroft et al., Clin Allergy 1977, 7, 235-243. NKA challenge is
performed by inhaling increasing concentrations of NKA as described
by Van Schoor et al., Eur Respir J 1998, 12, 17-23. The effect of a
treatment on airway responsiveness is determined using appropriate
statistical methods.
Description 8
Use of an Animal Model to Assess Efficacy in Treating Chronic
Obstructive Pulmonary Disease
[0525] An animal model using mice chronically exposed to cigarette
smoke can be used for assessing efficacy in treating emphysema
(see, e.g., Martorana et al., Am J Respir Crit Care Med 2005, 172,
848-835; and Cavarra et al., Am J Respir Crit Care Med 2001, 164,
886-890). Six-week old C57B1/6J male mice are used. In the acute
study, the mice are exposed either to room air or to the smoke of
five cigarettes for 20 minutes. In the chronic study, the mice are
exposed to either room air or to the smoke of three cigarettes/day,
for 5 days/week, for 7 months.
[0526] For the acute study, mice are divided into three groups of
40 animals each. These groups are then divided into four subgroups
of 10 mice each as follows: (1) no treatment/air-exposed; (2) no
treatment/smoke-exposed; (3) a first dose of test compound plus
smoke-exposed; and (4) a second dose of test compound. In the first
group, trolox equivalent antioxidant capacity is assessed at the
end of the exposure in bronchoalveolar lavage fluid. In the second
group, cytokines and chemokines are determined in bronchoalveolar
lavage fluid using a commercial cytokine panel at 4 hours; and in
the third group bronchoalveolar lavage fluid cell count is assessed
at 24 hours.
[0527] For the chronic study, five groups of animals are used: (1)
no treatment/air-exposed; (2) a first dose of a test compound plus
air-exposed; (3) no treatment/smoke-exposed; (4) a second dose of
the test compound plus smoke-exposed; and (5) the first dose of the
test compound plus smoke exposed. Seven months after chronic
exposure to room air or cigarette smoke, 5 to 12 animals from each
group are sacrificed and the lungs fixed intratracheally with
formalin. Lung volume is measured by water displacement. Lungs are
stained. Assessment of emphysema includes mean linear intercept and
internal surface area. The volume density of macrophages, marked
immunohistochemically with anti-mouse Mac-3 monoclonal antibodies
is determined by point counting. A mouse is considered to have
goblet cell metaplasia when at least one or more midsize
bronchi/lung showed a positive periodic acid-Schiff staining For
the determination of desmosine, fresh lungs are homogenized,
processed, and analyzed by high-pressure liquid chromatography.
Description 9
Animal Models for Assessing Therapeutic Efficacy of Morpholinoalkyl
Fumarates for Treating Parkinson's Disease
MPTP Induced Neurotoxicity
[0528] MPTP, or 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine is a
neurotoxin that produces a Parkinsonian syndrome in both humans and
experimental animals. Studies of the mechanism of MPTP
neurotoxicity show that it involves the generation of a major
metabolite, MPP.sup.+, formed by the activity of monoamine oxidase
on MPTP Inhibitors of monoamine oxidase block the neurotoxicity of
MPTP in both mice and primates. The specificity of the neurotoxic
effects of MPP.sup.+ for dopaminergic neurons appears to be due to
the uptake of MPP.sup.+ by the synaptic dopamine transporter.
Blockers of this transporter prevent MPP.sup.+ neurotoxicity.
MPP.sup.+ has been shown to be a relatively specific inhibitor of
mitochondrial complex I activity, binding to complex I at the
retenone binding site and impairing oxidative phosphorylation. In
vivo studies have shown that MPTP can deplete striatal ATP
concentrations in mice. It has been demonstrated that MPP.sup.+
administered intrastriatally to rats produces significant depletion
of ATP as well as increased lactate concentration confined to the
striatum at the site of the injections. Compounds that enhance ATP
production can protect against MPTP toxicity in mice.
[0529] A compound of the disclosure is administered to animals such
as mice or rats for three weeks before treatment with MPTP. MPTP is
administered at an appropriate dose, dosing interval, and mode of
administration for 1 week before sacrifice. Control groups receive
either normal saline or MPTP hydrochloride alone. Following
sacrifice the two striate are rapidly dissected and placed in
chilled 0.1 M perchloric acid. Tissue is subsequently sonicated and
aliquots analyzed for protein content using a fluorometer assay.
Dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic
acid (HVA) are also quantified. Concentrations of dopamine and
metabolites are expressed as nmol/mg protein.
[0530] Compounds of the disclosure that protect against DOPAC
depletion induced by MPTP, HVA, and/or dopamine depletion are
neuroprotective and therefore can be useful for the treatment of
Parkinson's disease.
Haloperidol-Induced Hypolocomotion
[0531] The ability of a compound to reverse the behavioral
depressant effects of dopamine antagonists, such as haloperidol, in
rodents is considered a valid method for screening drugs with
potential anti-Parkinsonian effects (Mandhane, et al., Eur. J.
Pharmacol 1997, 328, 135-141). Hence, the ability of compounds of
Formula (I) or Formula (II) to block haloperidol-induced deficits
in locomotor activity in mice can be used to assess both in vivo
and potential anti-Parkinsonian efficacy.
[0532] Mice used in the experiments are housed in a controlled
environment and allowed to acclimatize before experimental use. One
and one-half (1.5) hours before testing, mice are administered 0.2
mg/kg haloperidol, a dose that reduces baseline locomotor activity
by at least 50%. A test compound is administered 5-60 min prior to
testing. The animals are then placed individually into clean, clear
polycarbonate cages with a flat perforated lid. Horizontal
locomotor activity is determined by placing the cages within a
frame containing a 3.times.6 array of photocells interfaced to a
computer to tabulate beam interrupts. Mice are left undisturbed to
explore for 1 h, and the number of beam interruptions made during
this period serves as an indicator of locomotor activity, which is
compared with data for control animals for statistically
significant differences.
6-Hydroxydopamine Animal Model
[0533] The neurochemical deficits seen in Parkinson's disease can
be reproduced by local injection of the dopaminergic neurotoxin,
6-hydroxydopamine (6-OHDA) into brain regions containing either the
cell bodies or axonal fibers of the nigrostriatal neurons. By
unilaterally lesioning the nigrostriatal pathway on only one-side
of the brain, a behavioral asymmetry in movement inhibition is
observed. Although unilaterally-lesioned animals are still mobile
and capable of self-maintenance, the remaining dopamine-sensitive
neurons on the lesioned side become supersensitive to stimulation.
This is demonstrated by the observation that following systemic
administration of dopamine agonists, such as apomorphine, animals
show a pronounced rotation in a direction contralateral to the side
of lesioning. The ability of compounds to induce contralateral
rotations in 6-OHDA lesioned rats has been shown to be a sensitive
model to predict drug efficacy in the treatment of Parkinson's
disease.
[0534] Male Sprague-Dawley rats are housed in a controlled
environment and allowed to acclimatize before experimental use.
Fifteen minutes prior to surgery, animals are given an
intraperitoneal injection of the noradrenergic uptake inhibitor
desipramine (25 mg/kg) to prevent damage to nondopamine neurons.
Animals are then placed in an anesthetic chamber and anesthetized
using a mixture of oxygen and isoflurane. Once unconscious, the
animals are transferred to a stereotaxic frame, where anesthesia is
maintained through a mask. The top of the head is shaved and
sterilized using an iodine solution. Once dry, a 2 cm long incision
is made along the midline of the scalp and the skin refracted and
clipped back to expose the skull. A small hole is then drilled
through the skull above the injection site. In order to lesion the
nigrostriatal pathway, the injection cannula is slowly lowered to
position above the right medial forebrain bundle at -3.2 mm
anterior posterior, -1.5 mm medial lateral from the bregma, and to
a depth of 7.2 mm below the dura mater. Two minutes after lowering
the cannula, 6-OHDA is infused at a rate of 0.5 .mu.L/min over 4
min, to provide a final dose of 8 .mu.g. The cannula is left in
place for an additional 5 min to facilitate diffusion before being
slowly withdrawn. The skin is then sutured shut, the animal removed
from the sterereotaxic frame, and returned to its housing. The rats
are allowed to recover from surgery for two weeks before behavioral
testing.
[0535] Rotational behavior is measured using a rotameter system
having stainless steel bowls (45 cm dia.times.15 cm high) enclosed
in a transparent Plexiglas cover around the edge of the bowl and
extending to a height of 29 cm. To assess rotation, rats are placed
in a cloth jacket attached to a spring tether connected to an
optical rotameter positioned above the bowl, which assesses
movement to the left or right either as partial (45.degree.) or
full (360.degree.) rotations.
[0536] To reduce stress during administration of a test compound,
rats are initially habituated to the apparatus for 15 min on four
consecutive days. On the test day, rats are given a test compound,
e.g., a compound of Formula (I) or Formula (II). Immediately prior
to testing, animals are given a subcutaneous injection of a
sub-threshold dose of apomorphine, and then placed in the harness
and the number of rotations recorded for one hour. The total number
of full contralateral rotations during the hour test period serves
as an index of anti-Parkinsonian drug efficacy.
Description 10
Animal Model for Assessing Therapeutic Efficacy of Morpholinoalkyl
Fumarates for Treating Alzheimer's Disease
[0537] Heterozygous transgenic mice expressing the Swedish AD
mutant gene, hAPPK670N, M671L (Tg2576; Hsiao, Learning & Memory
2001, 8, 301-308) are used as an animal model of Alzheimer's
disease. Animals are housed under standard conditions with a 12:12
light/dark cycle and food and water available ad libitum. Beginning
at 9 months of age, mice are divided into three groups. The first
two groups of animals receive increasing doses of a compound of
Formula (I) or Formula (II), over six weeks. The remaining control
group receives daily saline injections for six weeks.
[0538] Behavioral testing is performed at each drug dose using the
same sequence over two weeks in all experimental groups: (1)
spatial reversal learning, (2) locomotion, (3) fear conditioning,
and (4) shock sensitivity.
[0539] Acquisition of the spatial learning paradigm and reversal
learning are tested during the first five days of test compound
administration using a water T-maze as described in Bardgett et
al., Brain Res Bull 2003, 60, 131-142. Mice are habituated to the
water T-maze during days 1-3, and task acquisition begins on day 4.
On day 4, mice are trained to find the escape platform in one
choice arm of the maze until 6 to 8 correct choices are made on
consecutive trails. The reversal learning phase is then conducted
on day 5. During the reversal learning phase, mice are trained to
find the escape platform in the choice arm opposite from the
location of the escape platform on day 4. The same performance
criteria and inter-trial interval are used as during task
acquisition.
[0540] Large ambulatory movements are assessed to determine that
the results of the spatial reversal learning paradigm are not
influenced by the capacity for ambulation. After a rest period of
two days, horizontal ambulatory movements, excluding vertical and
fine motor movements, are assessed in a chamber equipped with a
grid of motion-sensitive detectors on day 8. The number of
movements accompanied by simultaneous blocking and unblocking of a
detector in the horizontal dimension are measured during a one-hour
period.
[0541] The capacity of an animal for contextual and cued memory is
tested using a fear conditioning paradigm beginning on day 9.
Testing takes place in a chamber that contains a piece of absorbent
cotton soaked in an odor-emitting solution such as mint extract
placed below the grid floor. A 5-min, 3 trial 80 db, 2800 Hz
tone-foot shock sequence is administered to train the animals on
day 9. On day 10, memory for context is tested by returning each
mouse to the chamber without exposure to the tone and foot shock,
and recording the presence or absence of freezing behavior every 10
seconds for 8 minutes. Freezing is defined as no movement, such as
ambulation, sniffing or stereotypy, other than respiration.
[0542] On day 11, the response of the animal to an alternate
context and to the auditory cue is tested. Coconut extract is
placed in a cup and the 80 dB tone is presented, but no foot shock
is delivered. The presence or absence of freezing in response to
the alternate context is then determined during the first 2 minutes
of the trial. The tone is then presented continuously for the
remaining 8 minutes of the trial, and the presence or absence of
freezing in response to the tone is determined.
[0543] On day 12, the animals are tested to assess their
sensitivity to the conditioning stimulus, i.e., foot shock.
[0544] Following the last day of behavioral testing, animals are
anesthetized and the brains removed, post-fixed overnight, and
sections cut through the hippocampus. The sections are stained to
image .beta.-amyloid plaques.
[0545] Data is analyzed using appropriate statistical methods.
Description 11
Animal Model for Assessing Therapeutic Efficacy of Morpholinoalkyl
Fumarates for Treating Huntington's Disease
Neuroprotective Effects in a Transgenic Mouse Model of Huntington's
Disease
[0546] Transgenic HD mice of the N171-82Q strain and non-transgenic
littermates are treated with a compound of Formula (I), a compound
of Formula (II), or a vehicle from 10 weeks of age. The mice are
placed on a rotating rod ("rotarod"). The length of time at which a
mouse falls from the rotarod is recorded as a measure of motor
coordination. The total distance traveled by a mouse is also
recorded as a measure of overall locomotion. Mice administered
compounds of the disclosure that are neuroprotective in the
N171-82Q transgenic HD mouse model remain on the rotarod for a
longer period of time and travel farther than mice administered
vehicle.
Malonate Model of Huntington's Disease
[0547] A series of reversible and irreversible inhibitors of
enzymes involved in energy generating pathways has been used to
generate animal models for neurodegenerative diseases such as
Parkinson's and Huntington's diseases. In particular, inhibitors of
succinate dehydrogenase, an enzyme that impacts cellular energy
homeostasis, has been used to generate a model for Huntington's
disease.
[0548] To evaluate the effect of compounds of Formula (I) or
Formula (II) in this malonate model for Huntington's disease, a
compound of Formula (I) or Formula (II) is administered at an
appropriate dose, dosing interval, and route, to male
Sprague-Dawley rats. A compound of Formula (I) or Formula (II) is
administered for two weeks prior to the administration of malonate
and then for an additional week prior to sacrifice. Malonate is
dissolved in distilled deionized water and the pH adjusted to 7.4
with 0.1 M HCl. Intrastriatal injections of 1.5 .mu.L of 3 .mu.mol
malonate are made into the left striatum at the level of the
Bregma, 2.4 mm lateral to the midline and 4.5 mm ventral to the
dura. Animals are sacrificed at 7 days by decapitation and the
brains quickly removed and placed in ice cold 0.9% saline solution.
Brains are sectioned at 2 mm intervals in a brain mold. Slices are
then placed posterior side down in 2% 2,3,5-tiphenyltetrazolium
chloride. Slices are stained in the dark at room temperature for 30
min and then removed and placed in 4% paraformaldehyde pH 7.3.
Lesions, noted by pale staining, are evaluated on the posterior
surface of each section. The measurements are validated by
comparison with measurements obtained on adjacent Nissl stain
sections. Compounds exhibiting a neuroprotective effect and
therefore potentially useful in treating Huntington's disease show
a reduction in malonate-induced lesions.
Description 12
Animal Model for Assessing Therapeutic Efficacy of Morpholinoalkyl
Fumarates for Treating Amyotrophic Lateral Sclerosis
[0549] A murine model of SOD1 mutation-associated ALS has been
developed in which mice express the human superoxide dismutase
(SOD) mutation glycine.fwdarw.alanine at residue 93 (SOD1). These
SOD1 mice exhibit a dominant gain of the adverse property of SOD,
and develop motor neuron degeneration and dysfunction similar to
that of human ALS. The SOD1 transgenic mice show signs of posterior
limb weakness at about 3 months of age and die at 4 months.
Features common to human ALS include astrocytosis, microgliosis,
oxidative stress, increased levels of cyclooxygenase/prostaglandin,
and, as the disease progresses, profound motor neuron loss.
[0550] Studies are performed on transgenic mice overexpressing
human Cu/Zn-SOD G93A mutations (B6SJL-TgN (SOD1-G93A) 1 Gur) and
non-transgenic B6/SJL mice and their wild litter mates. Mice are
housed on a 12-hr day/light cycle and (beginning at 45 d of age)
allowed ad libitum access to either test compound-supplemented
chow, or, as a control, regular formula cold press chow processed
into identical pellets. Genotyping can be conducted at 21 days of
age as described in Gurney et al., Science 1994, 264(5166),
1772-1775. The SOD1 mice are separated into groups and treated with
a test compound, e.g., compound of Formula (I) or Formula (II), or
serve as controls.
[0551] The mice are observed daily and weighed weekly. To assess
health status mice are weighed weekly and examined for changes in
lacrimation/salivation, palpebral closure, ear twitch and pupillary
responses, whisker orienting, postural and righting reflexes and
overall body condition score. A general pathological examination is
conducted at the time of sacrifice.
[0552] Motor coordination performance of the animals can be
assessed by one or more methods known to those skilled in the art.
For example, motor coordination can be assessed using a
neurological scoring method. In neurological scoring, the
neurological score of each limb is monitored and recorded according
to a defined 4-point scale: 0--normal reflex on the hind limbs
(animal will splay its hind limbs when lifted by its tail);
1--abnormal reflex of hind limbs (lack of splaying of hind limbs
when animal is lifted by the tail); 2--abnormal reflex of limbs and
evidence of paralysis; 3--lack of reflex and complete paralysis;
and 4--inability to right when placed on the side in 30 seconds or
found dead. The primary end point is survival with secondary end
points of neurological score and body weight. Neurological score
observations and body weight are made and recorded five days per
week. Data analysis is performed using appropriate statistical
methods.
[0553] The rotarod test evaluates the ability of an animal to stay
on a rotating dowel allowing evaluation of motor coordination and
proprioceptive sensitivity. The apparatus is a 3 cm diameter
automated rod turning at, for example, 12 rounds per min. The
rotarod test measures how long the mouse can maintain itself on the
rod without falling. The test can be stopped after an arbitrary
limit, for example at 120 sec. If the animal falls down before 120
sec, the performance is recorded and two additional trials are
performed. The mean time of 3 trials is calculated. A motor deficit
is indicated by a decrease of walking time.
[0554] In the grid test, mice are placed on a grid (length: 37 cm,
width: 10.5 cm, mesh size: 1.times.1 cm.sup.2) situated above a
plane support. The number of times the mice put their paws through
the grid is counted and serves as a measure for motor
coordination.
[0555] The hanging test evaluates the ability of an animal to hang
on a wire. The apparatus is a wire stretched horizontally 40 cm
above a table. The animal is attached to the wire by its forepaws.
The time needed by the animal to catch the string with its hind
paws is recorded (60 sec max) during three consecutive trials.
[0556] Electrophysiological measurements (EMG) can also be used to
assess motor activity condition. Electromyographic recordings are
performed using an electromyography apparatus. During EMG
monitoring mice are anesthetized. The measured parameters are the
amplitude and the latency of the compound muscle action potential
(CMAP). CMAP is measured in gastrocnemius muscle after stimulation
of the sciatic nerve. A reference electrode is inserted near the
Achilles tendon and an active needle placed at the base of the
tail. A ground needle is inserted on the lower back of the mice.
The sciatic nerve is stimulated with a single 0.2 msec pulse at
supramaximal intensity (12.9 mA). The amplitude (mV) and the
latency of the response (ms) are measured. The amplitude is
indicative of the number of active motor units, while distal
latency reflects motor nerve conduction velocity.
[0557] The efficacy of test compounds can also be evaluated using
biomarker analysis. To assess the regulation of protein biomarkers
in SOD1 mice during the onset of motor impairment, samples of
lumbar spinal cord (protein extracts) are applied to ProteinChip
Arrays with varying surface chemical/biochemical properties and
analyzed, for example, by surface enhanced laser desorption
ionization time of flight mass spectrometry. Then, using integrated
protein mass profile analysis methods data is used to compare
protein expression profiles of the various treatment groups.
Analysis can be performed using appropriate statistical
methods.
Description 13
Animal Model for Assessing GI Irritation of Morpholinoalkyl
Fumarates
[0558] At least one MMF prodrug, e.g., dimethyl fumarate, is known
to cause gastrointestinal irritation. The Annamalai-Ma
gastrointestinal irritation rat model is predictive of
gastrointestinal irritation of MMF prodrugs in humans. This animal
model has several common features of other published GI irritation
animal models including the Whiteley-Dalrymple model described in
Models of Inflammation: Measuring Gastrointestinal Ulceration in
the Rat, Pharmacology (1998) 10.2.1-10.2.4; as well as the animal
models disclosed in Joseph J. Bertone, DVM, MS, DipACVIM.
Prevalence of Gastric Ulcers in Elite, Heavy Use Western
Performance Horses, AAEP Proceedings/Vol. 46/2000; and sbil
Buyukcoskun N., Central Effects of Glucagon-like Peptide-1 on Cold
Restraint Stress-induced Gastric Mucosal Lesions, Physiol. Res. 48:
451-455, 1999.
[0559] In order to assess gastrointestinal irritation using this
model, rats are treated orally with either vehicle or the MMF
prodrug of the present disclosure (n=10 per group) at 180
mg-equivalents MMF/kg of animal body weight, dosed once per day for
4 days, followed by necropsy and gastrointestinal evaluation at 24
hrs after the final dose. Evans Blue dye is injected IV (tail vein)
to visually emphasize any lesions in the gastrointestinal
tissue.
[0560] Accordingly, rats are dosed once per day for 4 consecutive
days with 180 mg-equivalents MMF/kg body weight per day. The
animals are fasted overnight prior to necropsy. On Day 5, to help
visualize lesions, 1 mL of 1% Evan's blue in saline is injected
into the tail vein 30 minutes prior to euthanasia. The animals are
euthanized by inhalation of carbon dioxide. A partial necropsy,
limited to the abdominal cavities, is then performed. The stomach
and small intestine are removed. Residual material is washed away,
using an irrigation syringe filled with saline. The stomach is cut
along the larger curvature and washed gently with normal saline,
and is examined for any lesions. The stomachs are scored in
accordance with the scoring system outlined in Table 4.
TABLE-US-00004 TABLE 4 Scoring System for Stomach Lesions in the
Rat Score Characteristics 0 Normal mucosa. 1 Non-erosive mucosal
changes. Swelling and reddening without any apparent mucosal
defect. 2 Apparent mucosal erosions. 3 Mild ulceration 1-5 small
lesions (1-2 mm). 4 Moderate ulceration: More than 5 small lesions
or 1 intermediate lesion (3-4 mm). 5 Severe ulceration: two or more
intermediate lesions or gross lesions (longer than 4 mm).
[0561] The GI irritation score values of the compounds tested are
set forth in Table 5, below and in FIG. 1. The GI data given below
clearly shows a several fold improvement over dimethyl fumarate
(DMF) and a Comparative Compound. Moreover, the representative
compounds of the disclosure showed normal mucosa or almost no
stomach irritation.
TABLE-US-00005 TABLE 5 GI Effect of Exemplary Compounds Compound
Stomach ID Structure Irritation Score 2 ##STR00120## 0 4
##STR00121## 0 6 ##STR00122## 0.6 8 ##STR00123## 0 10 ##STR00124##
0 CC-1 ##STR00125## 2.9 DMF ##STR00126## 4.6
[0562] Finally, it should be noted that there are alternative ways
of implementing the embodiments disclosed herein. Accordingly, the
present embodiments are to be considered as illustrative and not
restrictive, and the claims are not to be limited to the details
given herein, but may be modified within the scope and equivalents
thereof.
[0563] From the foregoing description, various modifications and
changes in the compositions and methods of this disclosure will
occur to those skilled in the art. All such modifications coming
within the scope of the appended claims are intended to be included
therein.
[0564] All publications, including but not limited to patents and
patent applications, cited in this specification are herein
incorporated by reference as if each individual publication were
specifically and individually indicated to be incorporated by
reference herein as though fully set forth.
[0565] At least some of the chemical names of compounds of the
disclosure as given and set forth in this application, may have
been generated on an automated basis by use of a commercially
available chemical naming software program, and have not been
independently verified. In the instance where the indicated
chemical name and the depicted structure differ, the depicted
structure will control.
[0566] Chemical structures shown herein were prepared using
ChemDraw or ISIS.RTM./DRAW. Any open valency appearing on a carbon,
oxygen or nitrogen atom in the structures herein indicates the
presence of a hydrogen atom. Where a chiral center exists in a
structure but no specific stereochemistry is shown for the chiral
center, both enantiomers associated with the chiral structure are
encompassed by the structure.
* * * * *