U.S. patent application number 14/946721 was filed with the patent office on 2016-05-26 for compounds for treating ophthalmic diseases and disorders.
The applicant listed for this patent is Acucela Inc.. Invention is credited to Jennifer GAGE, Feng HONG, Ryo KUBOTA, Vladimir A. KUKSA, Ian L. SCOTT.
Application Number | 20160145198 14/946721 |
Document ID | / |
Family ID | 41426558 |
Filed Date | 2016-05-26 |
United States Patent
Application |
20160145198 |
Kind Code |
A1 |
SCOTT; Ian L. ; et
al. |
May 26, 2016 |
COMPOUNDS FOR TREATING OPHTHALMIC DISEASES AND DISORDERS
Abstract
Provided are compounds, pharmaceutical compositions thereof, and
methods of treating ophthalmic diseases and disorders, such as
age-related macular degeneration and Stargardt's Disease, using
said compounds and compositions.
Inventors: |
SCOTT; Ian L.; (Dublin,
CA) ; KUKSA; Vladimir A.; (Bothell, WA) ;
HONG; Feng; (Bellevue, WA) ; KUBOTA; Ryo;
(Seattle, WA) ; GAGE; Jennifer; (Kenmore,
WA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Acucela Inc. |
Seattle |
WA |
US |
|
|
Family ID: |
41426558 |
Appl. No.: |
14/946721 |
Filed: |
November 19, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14175959 |
Feb 7, 2014 |
9193669 |
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14946721 |
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12603025 |
Oct 21, 2009 |
8674137 |
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14175959 |
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61197083 |
Oct 22, 2008 |
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61197082 |
Oct 22, 2008 |
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61197081 |
Oct 22, 2008 |
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61197091 |
Oct 22, 2008 |
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Current U.S.
Class: |
514/629 ;
514/649; 514/651; 514/653; 564/218; 564/223; 564/342; 564/346;
564/355 |
Current CPC
Class: |
C07C 215/68 20130101;
C07C 311/08 20130101; A61K 31/18 20130101; C07C 233/36 20130101;
C07C 2601/14 20170501; C07C 225/06 20130101; C07C 217/58 20130101;
C07B 2200/05 20130101; C07C 225/22 20130101; C07C 217/08 20130101;
C07C 211/52 20130101; C07C 217/10 20130101; A61P 9/00 20180101;
C07C 215/42 20130101; A61P 27/02 20180101; A61P 9/10 20180101; C07C
233/43 20130101; C07C 275/40 20130101; C07B 59/001 20130101; C07C
233/62 20130101; C07C 215/08 20130101; A61P 43/00 20180101; C07C
215/14 20130101; A61K 31/167 20130101; C07D 277/28 20130101; C07C
311/10 20130101; C07C 2601/08 20170501; C07C 311/14 20130101; C07C
233/25 20130101; C07C 311/21 20130101; C07C 211/49 20130101; C07C
335/18 20130101; C07C 311/24 20130101 |
International
Class: |
C07C 233/25 20060101
C07C233/25; C07C 225/06 20060101 C07C225/06; C07C 215/08 20060101
C07C215/08 |
Claims
1.-98. (canceled)
99. A compound of Formula (I) or tautomer, stereoisomer, geometric
isomer or a pharmaceutically acceptable salt or N-oxide thereof:
##STR00444## wherein, Z is
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--; G is selected from
--N(R.sup.42)C(.dbd.O)--R.sup.40, or
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40; R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18); each R.sup.42 is
hydrogen; R.sup.1 and R.sup.2 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl; or R.sup.1 and R.sup.2
together form an oxo; R.sup.3 and R.sup.4 are each hydrogen;
R.sup.9 is OR.sup.19; R.sup.10 is each is selected from hydrogen,
halogen, C.sub.1-C.sub.5 alkyl or --OR.sup.19; or R.sup.9 and
R.sup.10 form an oxo; R.sup.11 is hydrogen and R.sup.12 is
hydrogen, --CH.sub.3, or --C(.dbd.O)R.sup.23; R.sup.23 is
C.sub.1-C.sub.5 alkyl; R.sup.19 and R.sup.34 are each independently
hydrogen or C.sub.1-C.sub.5 alkyl; R.sup.16 and R.sup.17 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
halo; R.sup.18 is selected from hydrogen, C.sub.1-C.sub.5 alkyl,
alkoxy, hydroxy, halo or fluoroalkyl; each R.sup.33 is
independently selected from halogen, OR.sup.34, C.sub.1-C.sub.5
alkyl, or fluoroalkyl; and n is 0, 1, or 2.
100. The compound of claim 99 wherein, G is selected from
--N(R.sup.42)C(.dbd.O)--R.sup.40; and R.sup.10 is hydrogen.
101. The compound of claim 99 wherein, G is selected from
--N(R.sup.42)C(.dbd.O)--R.sup.40; and R.sup.9 and R.sup.10 form an
oxo.
102. The compound of claim 100 wherein, R.sup.11 and R.sup.12 are
each hydrogen; and n is 0.
103. The compound of claim 101 wherein, R.sup.11 and R.sup.12 are
each hydrogen; and n is 0.
104. The compound of claim 102 wherein, R.sup.16 and R.sup.17 are
each independently selected from hydrogen or C.sub.1-C.sub.5 alkyl;
and R.sup.18 is selected from hydrogen, C.sub.1-C.sub.5 alkyl,
alkoxy, or hydroxyl.
105. The compound of claim 103 wherein, R.sup.16 and R.sup.17 are
each independently selected from hydrogen, C.sub.1-C.sub.5 alkyl;
and R.sup.18 is selected from hydrogen, C.sub.1-C.sub.5 alkyl,
alkoxy, or hydroxyl.
106. The compound of claim 99 wherein, G is selected from
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40; and R.sup.10 is
hydrogen.
107. The compound of claim 99 wherein, G is selected from
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40; and R.sup.9 and
R.sup.10 form an oxo.
108. The compound of claim 106 wherein, R.sup.11 and R.sup.12 are
each hydrogen; and n is 0.
109. The compound of claim 100 wherein, R.sup.11 and R.sup.12 are
each hydrogen; and n is 0.
110. The compound of claim 108 wherein, R.sup.11 and R.sup.12 are
each hydrogen; and n is 0.
111. The compound of claim 109 wherein, R.sup.11 and R.sup.12 are
each hydrogen; and n is 0.
112. The compound of claim 110 wherein, R.sup.16 and R.sup.17 are
each independently selected from hydrogen, --C.sub.1-C.sub.5 alkyl;
and R.sup.18 is selected from hydrogen, C.sub.1-C.sub.5 alkyl,
alkoxy, or hydroxyl.
113. The compound of claim 111 wherein, R.sup.16 and R.sup.17 are
each independently selected from hydrogen, C.sub.1-C.sub.5 alkyl;
and R.sup.18 is selected from hydrogen, C.sub.1-C.sub.5 alkyl,
alkoxy, or hydroxyl.
114. A compound chosen from: ##STR00445## ##STR00446##
115. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and a compound of claim 99 or a tautomer,
stereoisomer, geometric isomer, pharmaceutically acceptable salt,
or N-oxide thereof.
116. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and a compound of claim 114 or a tautomer,
stereoisomer, geometric isomer, pharmaceutically acceptable salt,
or N-oxide thereof.
Description
CROSS-REFERENCE
[0001] This application is a divisional of U.S. Utility application
Ser. No. 12/603,025, filed Oct. 21, 2009, which claims the benefit
of U.S. Provisional Application No. 61/197,083, filed Oct. 22,
2008, U.S. Provisional Application No. 61/197,082, filed Oct. 22,
2008, U.S. Provisional Application No. 61/197,081, filed Oct. 22,
2008; and U.S. Provisional Application No. 61/197,091, filed Oct.
22, 2008, each of which is incorporated herein by reference in its
entirety.
BACKGROUND OF THE INVENTION
[0002] Neurodegenerative diseases, such as glaucoma, macular
degeneration, and Alzheimer's disease, affect millions of patients
throughout the world. Because the loss of quality of life
associated with these diseases is considerable, drug research and
development in this area is of great importance.
[0003] Macular degeneration affects between ten and fifteen million
patients in the United States, and it is the leading cause of
blindness in aging populations worldwide. Age-related macular
degeneration (AMD) affects central vision and causes the loss of
photoreceptor cells in the central part of retina called the
macula. Macular degeneration can be classified into two types:
dry-type and wet-type. The dry-form is more common than the wet;
about 90% of age-related macular degeneration patients are
diagnosed with the dry-form. The wet-form of the disease and
geographic atrophy, which is the end-stage phenotype of dry AMD,
causes the most serious vision loss. All patients who develop
wet-form AMD are believed to previously have developed dry-form AMD
for a prolonged period of time. The exact causes of age-related
macular degeneration are still unknown. The dry-form of AMD may
result from the senescence and thinning of macular tissues
associated with the deposition of pigment in the macular retinal
pigment epithelium. In wet AMD, new blood vessels grow beneath the
retina, form scar tissue, bleed, and leak fluid. The overlying
retina can be severely damaged, creating "blind" areas in the
central vision.
[0004] For the vast majority of patients who have the dry-form of
macular degeneration, no effective treatment is yet available.
Because the dry-form precedes development of the wet-form of
macular degeneration, therapeutic intervention to prevent or delay
disease progression in the dry-form AMD would benefit patients with
dry-form AMD and might reduce the incidence of the wet-form.
[0005] Decline of vision noticed by the patient or characteristic
features detected by an ophthalmologist during a routine eye exam
may be the first indicator of age-related macular degeneration. The
formation of "drusen," or membranous debris beneath the retinal
pigment epithelium of the macula is often the first physical sign
that AMD is developing. Late symptoms include the perceived
distortion of straight lines and, in advanced cases, a dark, blurry
area or area with absent vision appears in the center of vision;
and/or there may be color perception changes.
[0006] Different forms of genetically-linked macular degenerations
may also occur in younger patients. In other maculopathies, factors
in the disease are heredity, nutritional, traumatic, infection, or
other ecologic factors.
[0007] Glaucoma is a broad term used to describe a group of
diseases that causes a slowly progressive visual field loss,
usually asymptomatically. The lack of symptoms may lead to a
delayed diagnosis of glaucoma until the terminal stages of the
disease. The prevalence of glaucoma is estimated to be 2.2 million
in the United States, with about 120,000 cases of blindness
attributable to the condition. The disease is particularly
prevalent in Japan, which has four million reported cases. In many
parts of the world, treatment is less accessible than in the United
States and Japan, thus glaucoma ranks as a leading cause of
blindness worldwide. Even if subjects afflicted with glaucoma do
not become blind, their vision is often severely impaired.
[0008] The progressive loss of peripheral visual field in glaucoma
is caused by the death of ganglion cells in the retina. Ganglion
cells are a specific type of projection neuron that connects the
eye to the brain. Glaucoma is usually accompanied by an increase in
intraocular pressure. Current treatment includes use of drugs that
lower the intraocular pressure; however, contemporary methods to
lower the intraocular pressure are often insufficient to completely
stop disease progression. Ganglion cells are believed to be
susceptible to pressure and may suffer permanent degeneration prior
to the lowering of intraocular pressure. An increasing number of
cases of normal-tension glaucoma are observed in which ganglion
cells degenerate without an observed increase in the intraocular
pressure. Current glaucoma drugs only treat intraocular pressure
and are ineffective in preventing or reversing the degeneration of
ganglion cells.
[0009] Recent reports suggest that glaucoma is a neurodegenerative
disease, similar to Alzheimer's disease and Parkinson's disease in
the brain, except that it specifically affects retinal neurons. The
retinal neurons of the eye originate from diencephalon neurons of
the brain. Though retinal neurons are often mistakenly thought not
to be part of the brain, retinal cells are key components of the
central nervous system, interpreting the signals from the
light-sensing cells.
[0010] Alzheimer's disease (AD) is the most common form of dementia
among the elderly. Dementia is a brain disorder that seriously
affects a person's ability to carry out daily activities.
Alzheimer's is a disease that affects four million people in the
United States alone. It is characterized by a loss of nerve cells
in areas of the brain that are vital to memory and other mental
functions. Currently available drugs can ameliorate AD symptoms for
a relatively period of time, but no drugs are available that treat
the disease or completely stop the progressive decline in mental
function. Recent research suggests that glial cells that support
the neurons or nerve cells may have defects in AD sufferers, but
the cause of AD remains unknown. Individuals with AD seem to have a
higher incidence of glaucoma and age-related macular degeneration,
indicating that similar pathogenesis may underlie these
neurodegenerative diseases of the eye and brain. (See Giasson et
al., Free Radic. Biol. Med. 32:1264-75 (2002); Johnson et al.,
Proc. Natl. Acad. Sci. USA 99:11830-35 (2002); Dentchev et al.,
Mol. Vis. 9:184-90 (2003)).
[0011] Neuronal cell death underlies the pathology of these
diseases. Unfortunately, very few compositions and methods that
enhance retinal neuronal cell survival, particularly photoreceptor
cell survival, have been discovered. A need therefore exists to
identify and develop compositions that that can be used for
treatment and prophylaxis of a number of retinal diseases and
disorders that have neuronal cell death as a primary, or
associated, element in their pathogenesis.
[0012] In vertebrate photoreceptor cells, the irradiance of a
photon causes isomerization of 11-cis-retinylidene chromophore to
all-trans-retinylidene and uncoupling from the visual opsin
receptors. This photoisomerization triggers conformational changes
of opsins, which, in turn, initiate the biochemical chain of
reactions termed phototransduction (Filipek et al., Annu. Rev.
Physiol. 65:851-79 (2003)). Regeneration of the visual pigments
requires that the chromophore be converted back to the
11-cis-configuration in the processes collectively called the
retinoid (visual) cycle (see, e.g., McBee et al., Prog. Retin. Eye
Res. 20:469-52 (2001)). First, the chromophore is released from the
opsin and reduced in the photoreceptor by retinol dehydrogenases.
The product, all-trans-retinol, is trapped in the adjacent retinal
pigment epithelium (RPE) in the form of insoluble fatty acid esters
in subcellular structures known as retinosomes (Imanishi et al., J.
Cell Biol. 164:373-87 (2004)).
[0013] In Stargardt's disease (Allikmets et al., Nat. Genet.
15:236-46 (1997)), a disease associated with mutations in the ABCR
transporter that acts as a flippase, the accumulation of
all-trans-retinal may be responsible for the formation of a
lipofuscin pigment, A2E, which is toxic towards retinal pigment
epithelial cells and causes progressive retinal degeneration and,
consequently, loss of vision (Mata et al., Proc. Natl. Acad. Sci.
USA 97:7154-59 (2000); Weng et al., Cell 98:13-23 (1999)). Treating
patients with an inhibitor of retinol dehydrogenases, 13-cis-RA
(Isotretinoin, Accutane.RTM., Roche), has been considered as a
therapy that might prevent or slow the formation of A2E and might
have protective properties to maintain normal vision (Radu et al.,
Proc. Natl. Acad. Sci. USA 100:4742-47 (2003)). 13-cis-RA has been
used to slow the synthesis of 11-cis-retinal by inhibiting
11-cis-RDH (Law et al., Biochem. Biophys. Res. Commun. 161:825-9
(1989)), but its use can also be associated with significant night
blindness. Others have proposed that 13-cis-RA works to prevent
chromophore regeneration by binding RPE65, a protein essential for
the isomerization process in the eye (Gollapalli et al., Proc.
Natl. Acad. Sci. USA 101:10030-35 (2004)). Gollapalli et al.
reported that 13-cis-RA blocked the formation of A2E and suggested
that this treatment may inhibit lipofuscin accumulation and, thus,
delay either the onset of visual loss in Stargardt's disease or
age-related macular degeneration, which are both associated with
retinal pigment-associated lipofuscin accumulation. However,
blocking the retinoid cycle and forming unliganded opsin may result
in more severe consequences and worsening of the patient's
prognosis (see, e.g., Van Hooser et al., J. Biol. Chem.
277:19173-82 (2002); Woodruff et al., Nat. Genet. 35:158-164
(2003)). Failure of the chromophore to form may lead to progressive
retinal degeneration and may produce a phenotype similar to Leber
Congenital Amaurosis (LCA), is a very rare genetic condition
affecting children shortly after birth.
SUMMARY OF THE INVENTION
[0014] A need exists in the art for an effective treatment for
treating ophthalmic diseases or disorders resulting in ophthalmic
disfunction including those described above. In particular, there
exists a pressing need for compositions and methods for treating
Stargardt's disease and age-related macular degeneration (AMD)
without causing further unwanted side effects such as progressive
retinal degeneration, LCA-like conditions, night blindness, or
systemic vitamin A deficiency. A need also exists in the art for
effective treatments for other ophthalmic diseases and disorders
that adversely affect the retina.
[0015] In one embodiment is a compound of Formula (I) or tautomer,
stereoisomer, geometric isomer or a pharmaceutically acceptable
solvate, hydrate, salt, N-oxide or prodrug thereof:
##STR00001##
wherein, [0016] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0017] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0018] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0019] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0020] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0021] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0022] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0023] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0024] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0025] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0026] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0027] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0028] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0029]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0030] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl;
[0031] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and R.sup.35 are each
independently hydrogen or alkyl; [0032] R.sup.20 and R.sup.21 are
each independently selected from hydrogen, alkyl, carbocyclyl,
heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0033] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0034] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0035] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0036] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0037] In another embodiment is the compound of Formula (I)
wherein, [0038] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)-- [0039] X is --O--,
--S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0040] R.sup.1 and R.sup.2 are each
independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0041] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0042] R.sup.36 and R.sup.37 are each independently
selected from hydrogen, halogen, C.sub.1-C.sub.5 alkyl,
fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.36 and
R.sup.37 together form an oxo; [0043] R.sup.3 and R.sup.4 are each
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or C-attached heterocyclyl; or
R.sup.3 and R.sup.4 together with the carbon atom to which they are
attached, form a carbocyclyl or heterocyclyl; or R.sup.3 and
R.sup.4 together form an imino; [0044] R.sup.7 and R.sup.8 are each
independently selected from hydrogen, alkyl, carbocyclyl,
heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0045] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; [0046] R.sup.11 and R.sup.12 are each
independently selected from hydrogen, alkyl, carbocyclyl,
--C(.dbd.O)R.sup.23, SO.sub.2R.sup.23, CO.sub.2R.sup.23 or
SO.sub.2NR.sup.28R.sup.29; or R.sup.11 and R.sup.12, together with
the nitrogen atom to which they are attached, form an
N-heterocyclyl; [0047] each R.sup.13, R.sup.22 and R.sup.23 is
independently selected from alkyl, heteroalkyl, alkenyl, aryl,
aralkyl, carbocyclyl, heteroaryl or heterocyclyl; [0048] R.sup.6,
R.sup.19, R.sup.30, R.sup.34 and R.sup.35 are each independently
hydrogen or alkyl; [0049] R.sup.20 and R.sup.21 are each
independently selected from hydrogen, alkyl, carbocyclyl,
heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0050] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0051] each R.sup.33
is independently selected from halogen, OR.sup.34, alkyl, or
fluoroalkyl; and n is 0, 1, 2, 3, or 4.
[0052] In another embodiment is the compound of Formula (I) having
the structure of Formula (Ia)
##STR00002##
wherein, [0053] Z is --C(R.sup.9)(R.sup.10)--C)(R.sup.1)(R.sup.2)--
or --O--C(R.sup.31)(R.sup.32)--; [0054] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0055] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0056] R.sup.3 and R.sup.4 are each independently
selected from hydrogen or alkyl; or R.sup.3 and R.sup.4 together
form an imino; [0057] R.sup.7 and R.sup.8 are each independently
selected from hydrogen, alkyl, carbocyclyl or --C(.dbd.O)R.sup.13;
or R.sup.7 and R.sup.8, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0058] R.sup.9 and
R.sup.10 are each independently selected from hydrogen, halogen,
alkyl, fluoroalkyl, --OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl;
or R.sup.9 and R.sup.10 together form an oxo; or optionally,
R.sup.9 and R.sup.1 together form a direct bond to provide a double
bond; or optionally, R.sup.9 and R.sup.1 together form a direct
bond, and R.sup.10 and R.sup.2 together form a direct bond to
provide a triple bond; [0059] R.sup.11 and R.sup.12 are each
independently selected from hydrogen, alkyl, carbocyclyl or
--C(.dbd.O)R.sup.23; or [0060] R.sup.11 and R.sup.12, together with
the nitrogen atom to which they are attached, form an
N-heterocyclyl; and [0061] each R.sup.13, R.sup.22 and R.sup.23 is
independently selected from alkyl, alkenyl, aryl, aralkyl,
carbocyclyl, heteroaryl or heterocyclyl; [0062] R.sup.6, R.sup.19,
and R.sup.34 are each independently hydrogen or alkyl; [0063] each
R.sup.33 is independently selected from halogen, OR.sup.34, alkyl,
or fluoroalkyl; and n is 0, 1, 2, 3, or 4; [0064] R.sup.20 and
R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, --C(.dbd.O)R.sup.22; or [0065] R.sup.20 and R.sup.21,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; and [0066] each R.sup.24, R.sup.25, R.sup.26,
R.sup.27, R.sup.28 and R.sup.29 is independently selected from
hydrogen, alkyl, alkenyl, fluoroalkyl, aryl, heteroaryl,
carbocyclyl or heterocyclyl.
[0067] In another embodiment is the compound of Formula (Ia)
wherein, [0068] Z is --C(R.sup.9)(R.sup.10)--C)(R.sup.1)(R.sup.2)--
or --O--C(R.sup.31)(R.sup.32)--; [0069] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0070] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0071] R.sup.3 and R.sup.4 are each independently
selected from hydrogen or alkyl; or R.sup.3 and R.sup.4 together
form an imino; [0072] R.sup.7 and R.sup.8 are each independently
selected from hydrogen, alkyl, carbocyclyl or --C(.dbd.O)R.sup.13;
or R.sup.7 and R.sup.8, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0073] R.sup.9 and
R.sup.10 are each independently selected from hydrogen, halogen,
alkyl, fluoroalkyl, --OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl;
or R.sup.9 and R.sup.10 together form an oxo; [0074] R.sup.11 and
R.sup.12 are each independently selected from hydrogen, alkyl,
carbocyclyl or --C(.dbd.O)R.sup.23; or R.sup.11 and R.sup.12,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; and [0075] each R.sup.13, R.sup.22 and R.sup.23 is
independently selected from alkyl, alkenyl, aryl, aralkyl,
carbocyclyl, heteroaryl or heterocyclyl; [0076] R.sup.6, R.sup.19,
and R.sup.34 are each independently hydrogen or alkyl; [0077] each
R.sup.33 is independently selected from halogen, OR.sup.34, alkyl,
or fluoroalkyl; and n is 0, 1, 2, 3, or 4; [0078] R.sup.20 and
R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, --C(.dbd.O)R.sup.22; or R.sup.20 and R.sup.21,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; and [0079] each R.sup.24, R.sup.25, R.sup.26,
R.sup.27, R.sup.28 and R.sup.29 is independently selected from
hydrogen, alkyl, alkenyl, fluoroalkyl, aryl, heteroaryl,
carbocyclyl or heterocyclyl.
[0080] In another embodiment is the compound of Formula (Ia)
wherein, G is selected from --N(R.sup.42)--SO.sub.2--R.sup.40;
R.sup.40 is selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl,
or heteroaryl.
[0081] In another embodiment is the compound of Formula (Ia) having
the structure of Formula (Ib):
##STR00003##
wherein, [0082] Z is --C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--
or --O--C(R.sup.31)(R.sup.32)--; [0083] R.sup.40 is selected from
--C(R.sup.16)(R.sup.17)(R.sup.18); [0084] R.sup.16 and R.sup.17 are
each independently selected from hydrogen, alkyl, halo, aryl,
heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or R.sup.16 and
R.sup.17, together with the carbon to which they are attached form
a carbocyclyl or heterocycle; [0085] R.sup.18 is selected from
hydrogen, alkyl, alkoxy, hydroxy, halo or fluoroalkyl; [0086]
R.sup.1 and R.sup.2 are each independently selected from hydrogen,
halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6 or
--NR.sup.7R.sup.8; or R.sup.1 and R.sup.2 together form an oxo;
[0087] R.sup.31 and R.sup.32 are each independently selected from
hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl; [0088] R.sup.3 and
R.sup.4 are each independently selected from hydrogen or alkyl; or
R.sup.3 and R.sup.4 together form an imino; [0089] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl or --C(.dbd.O)R.sup.13; or R.sup.7 and R.sup.8,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; [0090] R.sup.9 and R.sup.10 are each independently
selected from hydrogen, halogen, alkyl, fluoroalkyl, --OR.sup.19,
NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and R.sup.10 together
form an oxo; or optionally, R.sup.9 and R.sup.1 together form a
direct bond to provide a double bond; or optionally, R.sup.9 and
R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0091]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl or --C(.dbd.O)R.sup.23; or R.sup.11
and R.sup.12, together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0092] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or heterocyclyl;
[0093] R.sup.6, R.sup.19, and R.sup.34 are each independently
hydrogen or alkyl; [0094] each R.sup.33 is independently selected
from halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2,
3, or 4; [0095] R.sup.20 and R.sup.21 are each independently
selected from hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.22; or
R.sup.20 and R.sup.21, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; and [0096] each
R.sup.24, R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl.
[0097] In another embodiment is the compound of Formula (Ib)
wherein, R.sup.9 and R.sup.10 are each independently selected from
hydrogen, halogen, alkyl, fluoroalkyl, --OR.sup.19,
--NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and R.sup.10
together form an oxo.
[0098] In another embodiment is the compound of Formula (Ib) having
the structure of Formula (Ic):
##STR00004##
wherein, [0099] R.sup.1 and R.sup.2 are each independently selected
from hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl,
--OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and R.sup.2 together
form an oxo; [0100] R.sup.3 and R.sup.4 are each independently
selected from hydrogen or alkyl; or R.sup.3 and R.sup.4 together
form an imino; [0101] R.sup.7 and R.sup.8 are each independently
selected from hydrogen, alkyl, carbocyclyl or --C(.dbd.O)R.sup.13;
or R.sup.7 and R.sup.8, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0102] R.sup.9 and
R.sup.10 are each independently selected from hydrogen, halogen,
alkyl, fluoroalkyl, --OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl;
or R.sup.9 and R.sup.10 together form an oxo; [0103] R.sup.11 and
R.sup.12 are each independently selected from hydrogen, alkyl,
carbocyclyl or --C(.dbd.O)R.sup.23; or R.sup.11 and R.sup.12,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; [0104] each R.sup.13, R.sup.22 and R.sup.23 is
independently selected from alkyl, alkenyl, aryl, aralkyl,
carbocyclyl, heteroaryl or heterocyclyl; [0105] R.sup.6, R.sup.19
and R.sup.34 are each independently hydrogen or alkyl; [0106]
R.sup.20 and R.sup.21 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.22; or R.sup.20 and
R.sup.21, together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0107] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0108] R.sup.16 and
R.sup.17 are each independently selected from hydrogen,
C.sub.1-C.sub.13 alkyl, halo or fluoroalkyl; or R.sup.16 and
R.sup.17, together with the carbon to which they are attached form
a carbocyclyl or heterocycle; [0109] each R.sup.33 is independently
selected from halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is
0, 1, 2, 3, or 4; and [0110] R.sup.18 is selected from a hydrogen,
alkyl, alkoxy, hydroxy, halo or fluoroalkyl.
[0111] In another embodiment is the compound of Formula (Ic)
wherein n is 0 and each of R.sup.11 and R.sup.12 is hydrogen. In a
further embodiment is the compound wherein each of R.sup.3,
R.sup.4, R.sup.14 and R.sup.15 is hydrogen. In a further embodiment
is the compound wherein, [0112] R.sup.1 and R.sup.2 are each
independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, or --OR.sup.6; [0113] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, or
--OR.sup.19; or R.sup.9 and R.sup.10 together form an oxo; [0114]
R.sup.6 and R.sup.19 are each independently hydrogen or alkyl;
[0115] R.sup.16 and R.sup.17, together with the carbon to which
they are attached form a carbocyclyl or heterocycle; and [0116]
R.sup.18 is selected from a hydrogen, alkoxy or hydroxy.
[0117] In a further embodiment is the compound wherein R.sup.16 and
R.sup.17, together with the carbon to which they are attached, form
a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or
cyclooctyl, and R.sup.18 is hydrogen or hydroxy.
[0118] In another embodiment is the compound of Formula (Ic),
wherein R.sup.1 is hydrogen and R.sup.12 is --C(.dbd.O)R.sup.23,
wherein R.sup.23 is alkyl. In further embodiment is the compound
wherein, [0119] R.sup.1 and R.sup.2 are each independently selected
from hydrogen, halogen, C.sub.1-C.sub.5 alkyl, or --OR.sup.6;
[0120] R.sup.9 and R.sup.10 are each independently selected from
hydrogen, halogen, alkyl, or --OR.sup.19; or R.sup.9 and R.sup.10
together form an oxo; [0121] R.sup.6 and R.sup.19 are each
independently selected from hydrogen or alkyl; [0122] R.sup.16 and
R.sup.17, together with the carbon atom to which they are attached,
form a carbocyclyl; and [0123] R.sup.18 is hydrogen, hydroxy or
alkoxy.
[0124] In a further embodiment is the compound wherein [0125] n is
0; [0126] R.sup.16 and R.sup.17, together with the carbon atom to
which they are attached, form a cyclopentyl, cyclohexyl or
cyclohexyl; and [0127] R.sup.18 is hydrogen or hydroxy.
[0128] In a further embodiment is the compound of Formula (Ic),
wherein [0129] R.sup.1 and R.sup.2 are each independently selected
from hydrogen, halogen, C.sub.1-C.sub.5 alkyl or --OR.sup.6; [0130]
R.sup.9 and R.sup.10 are each independently selected from hydrogen,
halogen, alkyl, or --OR.sup.19; or R.sup.9 and R.sup.10 together
form an oxo; [0131] R.sup.6 and R.sup.19 are each independently
hydrogen or alkyl; [0132] R.sup.16 and R.sup.17 is independently
selected from C.sub.1-C.sub.13 alkyl; and [0133] R.sup.18 is
hydrogen, hydroxy or alkoxy.
[0134] In another embodiment is the compound of Formula (Ib) having
the structure of Formula (Id):
##STR00005##
wherein, [0135] R.sup.31 and R.sup.32 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0136] R.sup.3 and R.sup.4 are each independently selected from
hydrogen or alkyl; or R.sup.3 and R.sup.4 together form an imino;
[0137] R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, or --C(.dbd.O)R.sup.23; or R.sup.11
and R.sup.12, together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; [0138] R.sup.23 is selected from
alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;
[0139] R.sup.14 and R.sup.15 are each independently selected from
hydrogen or alkyl; [0140] R.sup.16 and R.sup.17 are each
independently selected from hydrogen, C.sub.1-C.sub.13 alkyl, halo
or fluoroalkyl; or [0141] R.sup.16 and R.sup.17, together with the
carbon atom to which they are attached, form a carbocyclyl or
heterocycle; [0142] R.sup.18 is selected from a hydrogen, alkyl,
alkoxy, hydroxy, halo or fluoroalkyl; [0143] R.sup.34 is hydrogen
or alkyl; and [0144] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0145] In another embodiment is the compound wherein n is 0 and
each of R.sup.11 and R.sup.12 is hydrogen. In another embodiment is
the compound wherein each R.sup.3, R.sup.4, R.sup.14 and R.sup.15
is hydrogen. In another embodiment is the compound wherein,
R.sup.31 and R.sup.32 are each independently hydrogen, or
C.sub.1-C.sub.5 alkyl; R.sup.16 and R.sup.17, together with the
carbon atom to which they are attached, form a carbocyclyl; and
R.sup.18 is hydrogen, hydroxy, or alkoxy. In a further embodiment
is the compound wherein R.sup.16 and R.sup.17, together with the
carbon atom to which they are attached form a cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl and
R.sup.18 is hydrogen or hydroxy. In a further embodiment is the
compound wherein, R.sup.31 and R.sup.32 are each independently
selected from hydrogen, or C.sub.1-C.sub.5 alkyl; and R.sup.18 is
hydrogen, hydroxy or alkoxy. In a further embodiment is the
compound wherein, R.sup.31 and R.sup.32 are each independently
hydrogen, or C.sub.1-C.sub.5 alkyl; R.sup.6 and R.sup.19 are each
independently hydrogen or alkyl; R.sup.16 and R.sup.17 is
independently selected from C.sub.1-C.sub.13 alkyl; and R.sup.18 is
hydrogen, hydroxy or alkoxy.
[0146] In another embodiment is the compound of Formula (I)
wherein, Z is a bond, --X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; and X is --S--,
--S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--, --C(.dbd.O)--,
--C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--. In a further embodiment is the compound
wherein, G is selected from --N(R.sup.42)--SO.sub.2--R.sup.40; and
R.sup.40 is selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl,
or heteroaryl.
[0147] In an additional embodiment is the compound of Formula (I)
having the structure of Formula (Ie):
##STR00006##
wherein, [0148] X is --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--,
--N(R.sup.30)--, --C(.dbd.O)--, --C(.dbd.CH.sub.2)--,
--C(.dbd.N--NR.sup.35)--, or --C(.dbd.N--OR.sup.35)--; [0149]
R.sup.31 and R.sup.32 are each independently selected from
hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl; [0150] R.sup.3 and
R.sup.4 are each independently selected from hydrogen or alkyl; or
R.sup.3 and R.sup.4 together form an imino; [0151] R.sup.11 and
R.sup.12 are each independently selected from hydrogen, alkyl,
carbocyclyl, or --C(.dbd.O)R.sup.23; or R.sup.11 and R.sup.12,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; [0152] R.sup.23 is selected from alkyl, alkenyl,
aryl, carbocyclyl, heteroaryl or heterocyclyl; [0153] R.sup.16 and
R.sup.17 are each independently selected from hydrogen,
C.sub.1-C.sub.13 alkyl, halo or fluoroalkyl; or R.sup.16 and
R.sup.17, together with the carbon atom to which they are attached,
form a carbocyclyl or heterocycle; [0154] R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0155] R.sup.18
is selected from a hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0156] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0157] In a further embodiment is the compound of Formula (Ie)
wherein n is 0 and each R.sup.11 and R.sup.12 is hydrogen. In a
further embodiment is the compound wherein each R.sup.3, R.sup.4,
R.sup.14 and R.sup.15 is hydrogen. In a further embodiment is the
compound wherein, R.sup.31 and R.sup.32 are each independently
hydrogen, or C.sub.1-C.sub.5 alkyl; R.sup.16 and R.sup.17, together
with the carbon atom to which they are attached, form a carbocyclyl
or heterocycle; and R.sup.18 is hydrogen, hydroxy, or alkoxy. In a
further embodiment is the compound wherein R.sup.16 and R.sup.17,
together with the carbon atom to which they are attached form a
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or
cyclooctyl and R.sup.18 is hydrogen or hydroxy. In a further
embodiment is the compound wherein, R.sup.31 and R.sup.32 are each
independently selected from hydrogen, or C.sub.1-C.sub.5 alkyl;
R.sup.16 and R.sup.17 is independently selected from
C.sub.1-C.sub.13 alkyl; and R.sup.18 is hydrogen, hydroxy or
alkoxy.
[0158] In an additional embodiment is the compound of Formula (Ia)
wherein, G is selected from --N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40; R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; each R.sup.42 is independently selected from hydrogen
or alkyl. In a further embodiment is the compound wherein, G is
selected from --N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40; R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; each R.sup.42 is independently selected from hydrogen
or alkyl. In a further embodiment is the compound wherein, R.sup.42
is a hydrogen; R.sup.40 is selected from
--C(R.sup.16)(R.sup.17)(R.sup.18); R.sup.16 and R.sup.17, together
with the carbon atom to which they are attached, form a carbocyclyl
or heterocycle; and R.sup.18 is hydrogen, hydroxy, or alkoxy. In a
further embodiment is the compound wherein, R.sup.42 is a hydrogen;
R.sup.40 is selected from --C(R.sup.16)(R.sup.17)(R.sup.18);
R.sup.16 and R.sup.17, together with the carbon atom to which they
are attached, form a carbocyclyl or heterocycle; and R.sup.18 is
hydrogen, hydroxy, or alkoxy.
[0159] In another embodiment is the compound of Formula (Ia)
wherein, [0160] G is selected from
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0161] each
R.sup.43 is independently selected from hydrogen, alkyl,
cycloalkyl, aralkyl, C-attached heterocyclyl, aryl, or heteroaryl;
or two R.sup.43 groups, together with the nitrogen to which they
are attached, may form a heterocyclyl; [0162] each R.sup.42 is
independently selected from hydrogen or alkyl.
[0163] In another embodiment is the compound Formula (Ia) wherein,
[0164] G is selected from
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0165] each
R.sup.43 is independently selected from hydrogen, alkyl,
cycloalkyl, aralkyl, C-attached heterocyclyl, aryl, or heteroaryl;
or two R.sup.43 groups, together with the nitrogen to which they
are attached, may form a heterocyclyl; and [0166] R.sup.42 is
hydrogen.
[0167] In a further embodiment is the compound wherein, [0168] each
R.sup.43 is independently selected from hydrogen, alkyl,
cycloalkyl, aralkyl, C-attached heterocyclyl, aryl, or heteroaryl;
or two R.sup.43 groups, together with the nitrogen to which they
are attached, may form a heterocyclyl; and [0169] R.sup.42 is
hydrogen.
[0170] In a further embodiment is the compound wherein, [0171]
R.sup.40 is selected from --C(R.sup.16)(R.sup.17)(R.sup.18); [0172]
R.sup.16 and R.sup.17 are each independently selected from
hydrogen, alkyl, halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or
fluoroalkyl; or R.sup.16 and R.sup.17, together with the carbon to
which they are attached form a carbocyclyl or heterocycle; [0173]
R.sup.18 is selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl.
[0174] In another embodiment is the compound of Formula (I) wherein
one, more than one, or all of the non-exchangeable .sup.1H atoms
have been substituted with .sup.2H atoms.
[0175] In a specific embodiment, the compound of Formula (I) is
selected from the group consisting of:
##STR00007## ##STR00008## ##STR00009## ##STR00010## ##STR00011##
##STR00012## ##STR00013## ##STR00014## ##STR00015## ##STR00016##
##STR00017## ##STR00018##
[0176] In an additional embodiment is a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound of
Formula (I) or tautomer, stereoisomer, geometric isomer or a
pharmaceutically acceptable solvate, hydrate, salt, N-oxide or
prodrug thereof:
##STR00019##
wherein, [0177] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.35)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0178] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0179] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0180] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0181] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0182] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0183] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0184] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0185] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0186] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0187] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0188] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0189] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0190]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0191] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0192] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0193] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0194] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0195] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0196] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0197] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0198] In an additional embodiment is a method for treating an
ophthalmic disease or disorder in a subject, comprising
administering to the subject a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound of
Formula (I) or tautomer, stereoisomer, geometric isomer or a
pharmaceutically acceptable solvate, hydrate, salt, N-oxide or
prodrug thereof:
##STR00020##
wherein, [0199] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0200] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0201] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0202] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0203] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0204] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0205] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0206] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0207] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0208] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0209] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0210] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0211] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0212]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0213] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0214] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0215] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0216] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0217] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0218] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0219] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0220] In a further embodiment is the method wherein the ophthalmic
disease or disorder is a retinal disease or disorder. In an
additional embodiment is the method wherein the retinal disease or
disorder is age-related macular degeneration or Stargardt's macular
dystrophy. In an additional embodiment is the method wherein the
ophthalmic disease or disorder is selected from retinal detachment,
hemorrhagic retinopathy, retinitis pigmentosa, optic neuropathy,
inflammatory retinal disease, proliferative vitreoretinopathy,
retinal dystrophy, hereditary optic neuropathy, Sorsby's fundus
dystrophy, uveitis, a retinal injury, a retinal disorder associated
with Alzheimer's disease, a retinal disorder associated with
multiple sclerosis, a retinal disorder associated with Parkinson's
disease, a retinal disorder associated with viral infection, a
retinal disorder related to light overexposure, and a retinal
disorder associated with AIDS. In an additional embodiment is the
method wherein the ophthalmic disease or disorder is selected from
diabetic retinopathy, diabetic maculopathy, retinal blood vessel
occlusion, retinopathy of prematurity, or ischemia reperfusion
related retinal injury.
[0221] In an additional embodiment is the method of inhibiting at
least one visual cycle trans-cis isomerase in a cell comprising
contacting the cell with a compound of Formula (I) as described
herein, thereby inhibiting the at least one visual cycle trans-cis
isomerase. In a further embodiment is the method wherein the cell
is a retinal pigment epithelial (RPE) cell.
[0222] In a further embodiment is the method of inhibiting at least
one visual cycle trans-cis isomerase in a subject comprising
administering to the subject the pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound of
Formula (I) or tautomer, stereoisomer, geometric isomer or a
pharmaceutically acceptable solvate, hydrate, salt, N-oxide or
prodrug thereof:
##STR00021##
wherein, [0223] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0224] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0225] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0226] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0227] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0228] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0229] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0230] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0231] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0232] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0233] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0234] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0235] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0236]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0237] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0238] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0239] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0240] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0241] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0242] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0243] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0244] In a further embodiment is the method wherein the subject is
human. In a further embodiment is the method wherein accumulation
of lipofuscin pigment is inhibited in an eye of the subject. In a
further embodiment is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E). In a further
embodiment is the method wherein degeneration of a retinal cell is
inhibited. In a further embodiment is the method wherein the
retinal cell is a retinal neuronal cell. In a further embodiment is
the method wherein the retinal neuronal coil is a photoreceptor
cell, an amacrine cell, a horizontal cell, a ganglion cell, or a
bipolar cell. In a further embodiment is the method wherein the
retinal cell is a retinal pigment epithelial (RPE) cell.
[0245] In an additional embodiment is a compound that inhibits
11-cis-retinol production with an IC.sub.50 of about 1 .mu.M or
less when assayed in vitro, utilizing extract of cells that express
RPE65 and LRAT, wherein the extract further comprises CRALBP,
wherein the compound is stable in solution for at least about 1
week at room temperature. In an additional embodiment, the compound
is a non-retinoid compound. In a further embodiment is the
compound, wherein the compound inhibits 11-cis-retinol production
with an IC.sub.50 of about 0.1 .mu.M or less. In a further
embodiment is the compound, wherein the compound inhibits
11-cis-retinol production with an IC.sub.50 of about 0.01 .mu.M or
less.
[0246] In an additional embodiment is a non-retinoid compound that
inhibits an 11-cis-retinol producing isomerase reaction, wherein
said isomerase reaction occurs in RPE, and wherein said compound
has an ED.sub.50 value of 1 mg/kg or less when administered to a
subject. In a further embodiment is the non-retinoid compound
wherein the ED.sub.50 value is measured after administering a
single dose of the compound to said subject for about 2 hours or
longer.
[0247] In a further embodiment is the non-retinoid compound wherein
the structure of the non-retinoid compound corresponds to Formula
(I) or tautomer, stereoisomer, geometric isomer or a
pharmaceutically acceptable solvate, hydrate, salt, N-oxide or
prodrug thereof:
##STR00022##
wherein, [0248] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.3)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0249] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0250] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0251] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0252] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0253] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0254] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0255] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0256] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0257] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0258] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0259] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0260] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0261]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0262] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0263] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0264] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0265] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0266] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0267] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0268] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0269] In an additional embodiment is a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound
that inhibits 11-cis-retinol production with an IC.sub.50 of about
1 .mu.M or less when assayed in vitro, utilizing extract of cells
that express RPE65 and LRAT, wherein the extract further comprises
CRALBP, wherein the compound is stable in solution for at least
about 1 week at room temperature. In an additional embodiment is a
pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a non-retinoid compound that inhibits an 11-cis-retinol
producing isomerase reaction, wherein said isomerase reaction
occurs in RPE, and wherein said compound has an ED.sub.50 value of
1 mg/kg or less when administered to a subject.
[0270] In an additional embodiment is a method of modulating
chromophore flux in a retinoid cycle comprising introducing into a
subject a compound of Formula (I) as described herein. In a further
embodiment is the method resulting in a reduction of lipofuscin
pigment accumulated in an eye of the subject. In another embodiment
is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E). In yet another
embodiment is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E).
[0271] In an additional embodiment is a method of modulating
chromophore flux in a retinoid cycle comprising introducing into a
subject a compound that inhibits 11-cis-retinol production as
described herein. In a further embodiment is the method resulting
in a reduction of lipofuscin pigment accumulated in an eye of the
subject. In another embodiment is the method wherein the lipofuscin
pigment is N-retinylidene-N-retinyl-ethanolamine (A2E). In yet
another embodiment is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E).
[0272] In an additional embodiment is a method of modulating
chromophore flux in a retinoid cycle comprising introducing into a
subject a non-retinoid compound that inhibits an 11-cis-retinol
producing isomerase reaction as described herein. In a further
embodiment is the method resulting in a reduction of lipofuscin
pigment accumulated in an eye of the subject. In another embodiment
is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E). In yet another
embodiment is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E).
[0273] In an additional embodiment is a method for treating an
ophthalmic disease or disorder in a subject, comprising
administering to the subject a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound
that inhibits 11-cis-retinol production with an IC.sub.50 of about
1 .mu.M or less when assayed in vitro, utilizing extract of cells
that express RPE65 and LRAT, wherein the extract further comprises
CRALBP, wherein the compound is stable in solution for at least
about 1 week at room temperature. In a further embodiment is the
method wherein the ophthalmic disease or disorder is age-related
macular degeneration or Stargardt's macular dystrophy. In a further
embodiment is the method wherein the ophthalmic disease or disorder
is selected from retinal detachment, hemorrhagic retinopathy,
retinitis pigmentosa, cone-rod dystrophy, Sorsby's fundus
dystrophy, optic neuropathy, inflammatory retinal disease, diabetic
retinopathy, diabetic maculopathy, retinal blood vessel occlusion,
retinopathy of prematurity, or ischemia reperfusion related retinal
injury, proliferative vitreoretinopathy, retinal dystrophy,
hereditary optic neuropathy, Sorsby's fundus dystrophy, uveitis, a
retinal injury, a retinal disorder associated with Alzheimer's
disease, a retinal disorder associated with multiple sclerosis, a
retinal disorder associated with Parkinson's disease, a retinal
disorder associated with viral infection, a retinal disorder
related to light overexposure, myopia, and a retinal disorder
associated with AIDS. In a further embodiment is the method
resulting in a reduction of lipofuscin pigment accumulated in an
eye of the subject.
[0274] In an additional embodiment is a method for treating an
ophthalmic disease or disorder in a subject, comprising
administering to the subject a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a non-retinoid
compound that inhibits an 11-cis-retinol producing isomerase
reaction, wherein said isomerase reaction occurs in RPE, and
wherein said compound has an ED.sub.50 value of 1 mg/kg or less
when administered to a subject. In a further embodiment is the
method wherein the ophthalmic disease or disorder is age-related
macular degeneration or Stargardt's macular dystrophy. In a further
embodiment is the method wherein the ophthalmic disease or disorder
is selected from retinal detachment, hemorrhagic retinopathy,
retinitis pigmentosa, cone-rod dystrophy, Sorsby's fundus
dystrophy, optic neuropathy, inflammatory retinal disease, diabetic
retinopathy, diabetic maculopathy, retinal blood vessel occlusion,
retinopathy of prematurity, or ischemia reperfusion related retinal
injury, proliferative vitreoretinopathy, retinal dystrophy,
hereditary optic neuropathy, Sorsby's fundus dystrophy, uveitis, a
retinal injury, a retinal disorder associated with Alzheimer's
disease, a retinal disorder associated with multiple sclerosis, a
retinal disorder associated with Parkinson's disease, a retinal
disorder associated with viral infection, a retinal disorder
related to light overexposure, myopia, and a retinal disorder
associated with AIDS. In a further embodiment is the method
resulting in a reduction of lipofuscin pigment accumulated in an
eye of the subject.
[0275] In a further embodiment is a method of inhibiting dark
adaptation of a rod photoreceptor cell of the retina comprising
contacting the retina with a compound of Formula (I) as described
herein.
[0276] In a further embodiment is a method of inhibiting dark
adaptation of a rod photoreceptor cell of the retina comprising
contacting the retina with a compound that inhibits 11-cis-retinol
production as described herein.
[0277] In a further embodiment is a method of inhibiting dark
adaptation of a rod photoreceptor cell of the retina comprising
contacting the retina with a non-retinoid compound that inhibits an
11-cis-retinol producing isomerase reaction as described
herein.
[0278] In a further embodiment is a method of inhibiting
regeneration of rhodopsin in a rod photoreceptor cell of the retina
comprising contacting the retina with a compound of Formula (I) as
described herein.
[0279] In a further embodiment is a method of inhibiting
regeneration of rhodopsin in a rod photoreceptor cell of the retina
comprising contacting the retina with a compound that inhibits
11-cis-retinol production as described herein.
[0280] In a further embodiment is a method of inhibiting
regeneration of rhodopsin in a rod photoreceptor cell of the retina
comprising contacting the retina with a non-retinoid compound that
inhibits an 11-cis-retinol producing isomerase reaction as
described herein.
[0281] In a further embodiment is a method of reducing ischemia in
an eye of a subject comprising administering to the subject the
pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a compound of Formula (I) or tautomer, stereoisomer,
geometric isomer or a pharmaceutically acceptable solvate, hydrate,
salt, N-oxide or prodrug thereof:
##STR00023##
wherein, [0282] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0283] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0284] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0285] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0286] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0287] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0288] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0289] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0290] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0291] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0292] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0293] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0294] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0295]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0296] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0297] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0298] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0299] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0300] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0301] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0302] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0303] In another embodiment is a method of reducing ischemia in an
eye of a subject comprising administering to the subject a
pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a compound that inhibits 11-cis-retinol production with
an IC.sub.50 of about 1 .mu.M or less when assayed in vitro,
utilizing extract of cells that express RPE65 and LRAT, wherein the
extract further comprises CRALBP, wherein the compound is stable in
solution for at least about 1 week at room temperature. In a
further embodiment is the method wherein the pharmaceutical
composition is administered under conditions and at a time
sufficient to inhibit dark adaptation of a rod photoreceptor cell,
thereby reducing ischemia in the eye.
[0304] In another embodiment is a method of reducing ischemia in an
eye of a subject comprising administering to the subject a
pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a non-retinoid compound that inhibits an 11-cis-retinol
producing isomerase reaction, wherein said isomerase reaction
occurs in RPE, and wherein said compound has an ED.sub.50 value of
1 mg/kg or less when administered to a subject. In a further
embodiment is the method wherein the pharmaceutical composition is
administered under conditions and at a time sufficient to inhibit
dark adaptation of a rod photoreceptor cell, thereby reducing
ischemia in the eye.
[0305] In another embodiment is a method of inhibiting
neovascularization in the retina of an eye of a subject comprising
administering to the subject a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound
that inhibits 11-cis-retinol production with an IC.sub.50 of about
1 .mu.M or less when assayed in vitro, utilizing extract of cells
that express RPE65 and LRAT, wherein the extract further comprises
CRALBP, wherein the compound is stable in solution for at least
about 1 week at room temperature. In a further embodiment is the
method wherein the pharmaceutical composition is administered under
conditions and at a time sufficient to inhibit dark adaptation of a
rod photoreceptor cell, thereby inhibiting neovascularization in
the retina.
[0306] In another embodiment is a method of inhibiting
neovascularization in the retina of an eye of a subject comprising
administering to the subject a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a non-retinoid
compound that inhibits an 11-cis-retinol producing isomerase
reaction, wherein said isomerase reaction occurs in RPE, and
wherein said compound has an ED.sub.50 value of 1 mg/kg or less
when administered to a subject. In a further embodiment is the
method wherein the pharmaceutical composition is administered under
conditions and at a time sufficient to inhibit dark adaptation of a
rod photoreceptor cell, thereby inhibiting neovascularization in
the retina.
[0307] In another embodiment is a method of inhibiting degeneration
of a retinal cell in a retina comprising contacting the retina with
the compound of Formula (I) as described herein. In a further
embodiment is the method wherein the retinal cell is a retinal
neuronal cell. In yet another embodiment is the method wherein the
retinal neuronal cell is a photoreceptor cell.
[0308] In another embodiment is a method of inhibiting degeneration
of a retinal cell in a retina comprising contacting the retina with
a compound that inhibits 11-cis-retinol production with an
IC.sub.50 of about 1 .mu.M or less when assayed in vitro, utilizing
extract of cells that express RPE65 and LRAT, wherein the extract
further comprises CRALBP, wherein the compound is stable in
solution for at least about 1 week at room temperature. In a
further embodiment is the method wherein the retinal cell is a
retinal neuronal cell. In yet another embodiment is the method
wherein the retinal neuronal cell is a photoreceptor cell.
[0309] In another embodiment is a method of inhibiting degeneration
of a retinal cell in a retina comprising contacting the retina with
a non-retinoid compound that inhibits an 11-cis-retinol producing
isomerase reaction, wherein said isomerase reaction occurs in RPE,
and wherein said compound has an ED.sub.50 value of 1 mg/kg or less
when administered to a subject. In a further embodiment is the
method wherein the retinal cell is a retinal neuronal cell. In yet
another embodiment is the method wherein the retinal neuronal cell
is a photoreceptor cell.
[0310] In a further embodiment is a method of reducing lipofuscin
pigment accumulated in a subject's retina comprising administering
to the subject a pharmaceutical composition comprising a
pharmaceutically acceptable carrier and a compound of Formula (I)
or tautomer, stereoisomer, geometric isomer or a pharmaceutically
acceptable solvate, hydrate, salt, N-oxide or prodrug thereof:
##STR00024##
wherein, [0311] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.3)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0312] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0313] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0314] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0315] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0316] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0317] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0318] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0319] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0320] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0321] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0322] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0323] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0324]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0325] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0326] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0327] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0328] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0329] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0330] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0331] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0332] In a further embodiment is the method wherein the lipofuscin
is N-retinylidene-N-retinyl-ethanolamine (A2E).
[0333] In another embodiment is a method of reducing lipofuscin
pigment accumulated in a subject's retina comprising administering
to the subject a pharmaceutical composition comprising a
pharmaceutically acceptable carrier and a compound that inhibits
11-cis-retinol production with an IC.sub.50 of about 1 .mu.M or
less when assayed in vitro, utilizing extract of cells that express
RPE65 and LRAT, wherein the extract further comprises CRALBP,
wherein the compound is stable in solution for at least about 1
week at room temperature. In a further embodiment is the method
wherein the lipofuscin is N-retinylidene-N-retinyl-ethanolamine
(A2E).
[0334] In another embodiment is a method of reducing lipofuscin
pigment accumulated in a subject's retina comprising administering
to the subject a pharmaceutical composition comprising a
pharmaceutically acceptable carrier and a non-retinoid compound
that inhibits an 11-cis-retinol producing isomerase reaction,
wherein said isomerase reaction occurs in RPE, and wherein said
compound has an ED.sub.50 value of 1 mg/kg or less when
administered to a subject. In a further embodiment is the method
wherein the lipofuscin is N-retinylidene-N-retinyl-ethanolamine
(A2E).
[0335] In one embodiment is a compound having a structure of
Formula (II):
##STR00025##
as an isolated E or Z geometric isomer or a mixture of E and Z
geometric isomers, as a tautomer or a mixture of tautomers, as a
stereoisomer or as a pharmaceutically acceptable salt, hydrate,
solvate, N-oxide or prodrug thereof, wherein: [0336] R.sup.1 and
R.sup.2 are each the same or different and independently hydrogen
or alkyl; [0337] R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are each the
same or different and independently hydrogen, halogen, nitro,
--NH.sub.2, --NHR.sup.13, --N(R.sup.13).sub.2, --OR.sup.12, alkyl
or fluoroalkyl; [0338] R.sup.7 and R.sup.8 are each the same or
different and independently hydrogen or alkyl; or R.sup.7 and
R.sup.8 together with the carbon atom to which they are attached,
form a carbocyclyl or heterocyclyl; or R.sup.7 and R.sup.8 together
form an imino; [0339] R.sup.9 is hydrogen, alkyl, carbocyclyl,
heterocyclyl, --C(.dbd.O)R.sup.13, --SO.sub.2R.sup.13,
--CO.sub.2R.sup.13, --CONH.sub.2, --CON(R.sup.13).sub.2 or
--CON(H)R.sup.13; [0340] R.sup.10 is hydrogen or alkyl; or R.sup.9
and R.sup.10, together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; [0341] R.sup.11 is alkyl,
alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl; [0342] each
R.sup.12 is independently selected from hydrogen or alkyl; [0343]
each R.sup.13 is independently selected from alkyl, carbocyclyl,
heterocyclyl, aryl or heteroaryl; [0344] Z is a bond, Y or W--Y,
wherein [0345] W is --C(R.sup.14)(R.sup.15)--, --O--, --S--,
--S(.dbd.O)--, --S(.dbd.O).sub.2-- or --N(R.sup.12)_; [0346] Y is
--C(R.sup.16)(R.sup.17)-- or
--C(R.sup.16)(R.sup.7)--C(R.sup.21)(R.sup.22)--; [0347] R.sup.14
and R.sup.15 are each the same or different and independently
hydrogen, halogen, alkyl, fluoroalkyl, --OR.sup.12,
--NR.sup.18R.sup.19, carbocyclyl or heterocyclyl; or R.sup.14 and
R.sup.15 together form an oxo, an imino, an oximo, or a hydrazino;
[0348] R.sup.16 and R.sup.17 are each the same or different and
independently hydrogen, halogen, alkyl, fluoroalkyl, --OR.sup.12,
--NR.sup.18R.sup.19, carbocyclyl or heterocyclyl; or R.sup.16 and
R.sup.17 together form an oxo; or [0349] optionally, R.sup.14 and
R.sup.16 together form a direct bond to provide a double bond
connecting W and Y; or optionally, R.sup.14 and R.sup.16 together
form a direct bond, and R.sup.15 and R.sup.17 together form a
direct bond to provide a triple bond connecting W and Y; [0350]
each R.sup.18 and R.sup.19 is independently selected from hydrogen,
alkyl, carbocyclyl, or --C(.dbd.O)R.sup.13, --SO.sub.2R.sup.13,
--CO.sub.2R.sup.13, --CONH.sub.2, --CON(R.sup.13).sub.2 or
--CON(H)R.sup.13; or R.sup.18 and R.sup.19, together with the
nitrogen atom to which they are attached, form an N-heterocyclyl;
[0351] R.sup.21 and R.sup.22 are each the same or different and
independently hydrogen, halogen, alkyl, fluoroalkyl, --OR.sup.12,
--NR.sup.18R.sup.19, carbocyclyl or heterocyclyl; [0352] provided
that when R.sup.11 is phenyl, the compound of Formula (A) is not:
[0353]
2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]ace-
tamide; [0354]
(2S,3R)-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)-e-
thenyl]phenyl]-butanamide; [0355] L-glutamic acid,
1-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]ester;
glycine, 3-hydroxy-5-[(1E)-2-(4-hydroxyphenyl)ethenyl]phenyl ester;
[0356]
(2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)etheny-
l]phenyl]propanamide; [0357]
(2S)-2-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)eth-
enyl]phenyl]propanamide; [0358] (2S)-2-amino-N-[2-methoxy-5-[(1
Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-4-methyl-pentanamide;
[0359] (2S)-2-amino-N-[2-methoxy-5-[(1
Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-3-methyl-butanamide;
or [0360]
2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phe-
nylbutanamide; and wherein the compound of Formula (II) is
isotopically enriched.
[0361] In another embodiment is the compound of Formula (II) having
one, more than one, or all of the non exchangeable .sup.1H atoms
are replaced with .sup.2H atoms.
[0362] In another embodiment is the compound of Formula (II) having
the structure of Formula (IIa):
##STR00026##
wherein R.sup.11 is selected from:
##STR00027##
and one, more than one, or all of the non-exchangeable .sup.1H
atoms are replaced with .sup.2H atoms.
[0363] In another embodiment is the compound of Formula (IIa)
selected from:
##STR00028##
[0364] One embodiment provides a compound having a structure of
Formula (III):
##STR00029## [0365] as a tautomer or a mixture of tautomers, or as
a pharmaceutically acceptable salt, hydrate, solvate, N-oxide,
stereoisomer, geometric isomer or prodrug thereof, wherein: [0366]
m is 0, 1, 2 or 3; [0367] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.21)(R.sup.22)--,
--C(R.sup.23)(R.sup.24)--C(R.sup.1)(R.sup.2)--, or
--C(R.sup.23)(R.sup.24)--C(R.sup.25)(R.sup.26)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.21)(R.sup.22)--C(R.sup.1)(R.sup.2)--,
--C(R.sup.32)(R.sup.33)--X--C(R.sup.21)(R.sup.22)--; [0368] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.31)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0369] Y is a bond,
--C(R.sup.27)(R.sup.28)--, or
--C(R.sup.27)(R.sup.2)--C(R.sup.29)(R.sup.30)--; [0370] R.sup.1 and
R.sup.2 are each independently selected from hydrogen, halogen,
C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6 or
--NR.sup.7R.sup.8; or R.sup.1 and R.sup.2 together form an oxo;
[0371] R.sup.21, R.sup.22, R.sup.32 and R.sup.33 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0372] R.sup.23 and R.sup.24 are each independently
selected from hydrogen, halogen, C.sub.1-C.sub.5 alkyl,
fluoroalkyl, --OR.sup.6, --NR.sup.7R.sup.8; or R.sup.23 and
R.sup.24 together form an oxo; or optionally, R.sup.23 and an
adjacent R.sup.1 together form a direct bond to provide a double
bond; or optionally, R.sup.23 and an adjacent R.sup.1 together form
a direct bond, and R.sup.24 and an adjacent R.sup.2 together form a
direct bond to provide a triple bond; [0373] R.sup.25 and R.sup.26
are each independently selected from hydrogen, halogen,
C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6 or
--NR.sup.7R.sup.8; or R.sup.25 and R.sup.26 together form an oxo;
[0374] R.sup.3 and R.sup.4 are each independently selected from
hydrogen, alkyl, heteroalkyl, alkenyl, fluoroalkyl, aryl,
heteroaryl, carbocyclyl or C-attached heterocyclyl; or R.sup.3 and
R.sup.4 together with the carbon atom to which they are attached,
form a carbocyclyl or heterocyclyl; or R.sup.3 and R.sup.4 together
form an imino; [0375] R.sup.5 is alkyl, heteroalkyl, alkenyl,
heteroalkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;
[0376] each R.sup.6 is the same or different and independently
hydrogen or C.sub.1-C.sub.5 alkyl; [0377] each R.sup.7 and each
R.sup.8 are each the same or different and independently hydrogen,
alkyl, carbocyclyl, heteroalkyl, heterocycloalkyl, aryl,
heteroaryl, --C(.dbd.O)R.sup.9, SO.sub.2R.sup.9, CO.sub.2R.sup.9,
SO.sub.2NH.sub.2, SO.sub.2NHR.sup.9 or SO.sub.2N(R.sup.9).sub.2; or
R.sup.7 and R.sup.8, together with the nitrogen atom to which they
are attached, form an N-heterocyclyl; [0378] each R.sup.9 is the
same or different and each is independently alkyl, alkenyl, aryl,
carbocyclyl, heteroaryl or heterocyclyl; [0379] R.sup.12 and
R.sup.13 are the same or different and independently hydrogen,
alkyl, heteroalkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl,
--C(.dbd.O)R.sup.9, SO.sub.2R.sup.9, CO.sub.2R.sup.9,
SO.sub.2NH.sub.2, SO.sub.2NHR.sup.9 or SO.sub.2N(R.sup.9).sub.2; or
R.sup.12 and R.sup.13 together with the nitrogen atom to which they
are attached, form an N-heterocyclyl; and [0380] each R.sup.14 is
the same or different and independently alkyl, halo, fluoroalkyl or
--OR.sup.6; [0381] each R.sup.27, R.sup.28, R.sup.29 and R.sup.31
are the same or different and independently hydrogen, alkyl or
--OR.sup.6; and [0382] R.sup.30 and R.sup.35 are each independently
hydrogen or C.sub.1-C.sub.5 alkyl; and wherein the compound of
Formula (III) is isotopically enriched.
[0383] Another embodiment provides the compound of Formula (III)
having one, more than one or all of the non-exchangeable .sup.1H
atoms replaced with .sup.2H atoms.
[0384] Another embodiment provides the compound of Formula
(IIIa):
##STR00030##
wherein Y is a bond; [0385] R.sup.5 is selected from:
[0385] ##STR00031## [0386] one, more than one, or all of the
non-exchangeable .sup.1H atoms are replaced with .sup.2H atoms.
[0387] Another embodiment provides the compound of Formula (III)
selected from:
##STR00032##
[0388] One embodiment provides a compound of Formula (IV) or
tautomer, stereoisomer, geometric isomer or a pharmaceutically
acceptable solvate, hydrate, salt, N-oxide or prodrug thereof:
##STR00033##
wherein, [0389] Z is --C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--
or --X--C(R.sup.3)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0390] R.sup.1
and R.sup.2 are each independently selected from hydrogen, halogen,
C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6 or
--NR.sup.7R.sup.8; or R.sup.1 and R.sup.2 together form an oxo;
[0391] R.sup.31 and R.sup.32 are each independently selected from
hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl; [0392] R.sup.36
and R.sup.37 are each independently selected from hydrogen,
halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6 or
--NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an oxo;
or optionally, R.sup.36 and R.sup.1 together form a direct bond to
provide a double bond; or optionally, R.sup.36 and R.sup.1 together
form a direct bond, and R.sup.37 and R.sup.2 together form a direct
bond to provide a triple bond; [0393] R.sup.3 and R.sup.4 are each
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or C-attached heterocyclyl; or
R.sup.3 and R.sup.4 together with the carbon atom to which they are
attached, form a carbocyclyl or heterocyclyl; or R.sup.3 and
R.sup.4 together form an imino; [0394] R.sup.5 is C.sub.5-C.sub.15
alkyl or carbocyclyalkyl; [0395] R.sup.7 and R.sup.8 are each
independently selected from hydrogen, alkyl, carbocyclyl,
heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0396] X is --O--, --S--, --S(.dbd.O)--,
--S(.dbd.O).sub.2--, --N(R.sup.30)--, --C(.dbd.O)--,
--C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0397] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0398]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0399] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0400] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0401] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0402] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0403] each R.sup.33
is independently selected from halogen, OR.sup.34, alkyl, or
fluoroalkyl; and n is 0, 1, 2, 3, or 4; with the provision that
R.sup.5 is not 2-(cyclopropyl)-1-ethyl or an unsubstituted normal
alkyl; and wherein [0404] the compound of Formula (IV) is
isotopically enriched.
[0405] Another embodiment provides the compound of Formula (IV) has
one, more than one or all of the non-exchangeable .sup.1H atoms
replaced with .sup.2H atoms.
[0406] Another embodiment provides the compound having the
structure of Formula (IVa):
##STR00034##
wherein Y is a bond; [0407] R.sup.5 is selected from:
##STR00035##
[0407] and one, more than one, or all of the non-exchangeable
.sup.1H atoms are replaced with .sup.2H atoms.
[0408] Another embodiment provides the compound selected from:
##STR00036##
[0409] One embodiment provides a method for treating an ophthalmic
disease or disorder in a subject, comprising administering to the
subject a compound of Formula (II), (IIa), (III), (IIIa), (IV), or
(IVa) as described herein, or tautomer, stereoisomer, geometric
isomer or a pharmaceutically acceptable solvate, hydrate, salt,
N-oxide or prodrug thereof. Another embodiment provides a method
for treating an ophthalmic disease or disorder wherein the
ophthalmic disease or disorder is age-related macular degeneration
or Stargardt's macular dystrophy.
INCORPORATION BY REFERENCE
[0410] All publications, patents, and patent applications mentioned
in this specification are herein incorporated by reference to the
same extent as if each individual publication, patent, or patent
application was specifically and individually indicated to be
incorporated by reference.
BRIEF DESCRIPTION OF THE DRAWINGS
[0411] The novel features of the invention are set forth with
particularity in the appended claims. A better understanding of the
features and advantages of the present invention will be obtained
by reference to the following detailed description that sets forth
illustrative embodiments, in which the principles of the invention
are utilized, and the accompanying drawings of which:
[0412] FIG. 1 depicts dose-dependent inhibition of 11-cis-retinol
production (as assayed by a human in vitro isomerase assay) by the
compound of Example 5 (Compound 5).
[0413] FIG. 2 depicts dose-dependent inhibition of 11-cis-retinol
production (as assayed by a human in vitro isomerase assay) by the
compound of Example 6 (Compound 6).
DETAILED DESCRIPTION OF THE INVENTION
[0414] Compounds are described herein that inhibit an isomerization
step of the retinoid cycle. These compounds and compositions
comprising these compounds are useful for inhibiting degeneration
of retinal cells or for enhancing retinal cell survival. The
compounds described herein are, therefore, useful for treating
ophthalmic diseases and disorders, including retinal diseases or
disorders, such as age related macular degeneration and Stargardt's
disease.
Nitrogen-Linked Compounds
[0415] In one embodiment is a compound of Formula (I) or tautomer,
stereoisomer, geometric isomer or a pharmaceutically acceptable
solvate, hydrate, salt, N-oxide or prodrug thereof:
##STR00037##
wherein, [0416] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0417] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0418] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0419] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0420] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0421] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0422] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0423] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0424] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0425] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0426] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0427] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0428] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0429]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0430] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0431] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0432] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0433] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0434] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0435] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0436] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0437] In another embodiment is the compound of Formula (I)
wherein, [0438] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--
or --X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0439] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0440] R.sup.1 and R.sup.2 are each
independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0441] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0442] R.sup.36 and R.sup.37 are each independently
selected from hydrogen, halogen, C.sub.1-C.sub.5 alkyl,
fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.36 and
R.sup.37 together form an oxo; [0443] R.sup.3 and R.sup.4 are each
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or C-attached heterocyclyl; or
R.sup.3 and R.sup.4 together with the carbon atom to which they are
attached, form a carbocyclyl or heterocyclyl; or R.sup.3 and
R.sup.4 together form an imino; [0444] R.sup.7 and R.sup.8 are each
independently selected from hydrogen, alkyl, carbocyclyl,
heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0445] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; [0446] R.sup.11 and R.sup.12 are each
independently selected from hydrogen, alkyl, carbocyclyl,
--C(.dbd.O)R.sup.23, SO.sub.2R.sup.23, CO.sub.2R.sup.23 or
SO.sub.2NR.sup.28R.sup.29; or R.sup.11 and R.sup.12, together with
the nitrogen atom to which they are attached, form an
N-heterocyclyl; [0447] each R.sup.13, R.sup.22 and R.sup.23 is
independently selected from alkyl, heteroalkyl, alkenyl, aryl,
aralkyl, carbocyclyl, heteroaryl or heterocyclyl; [0448] R.sup.6,
R.sup.19, R.sup.30, R.sup.34 and R.sup.35 are each independently
hydrogen or alkyl; [0449] R.sup.20 and R.sup.21 are each
independently selected from hydrogen, alkyl, carbocyclyl,
heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and each R.sup.24, R.sup.25,
R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is independently selected
from hydrogen, alkyl, alkenyl, fluoroalkyl, aryl, heteroaryl,
carbocyclyl or heterocyclyl; [0450] each R.sup.33 is independently
selected from halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is
0, 1, 2, 3, or 4.
[0451] In another embodiment is the compound of Formula (I) having
the structure of Formula (Ia)
##STR00038##
wherein, [0452] Z is --C(R.sup.9)(R.sup.10)--C)(R.sup.1)R.sup.2)--
or --O--C(R.sup.31)(R.sup.32)--; [0453] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0454] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0455] R.sup.3 and R.sup.4 are each independently
selected from hydrogen or alkyl; or R.sup.3 and R.sup.4 together
form an imino; [0456] R.sup.7 and R.sup.8 are each independently
selected from hydrogen, alkyl, carbocyclyl or --C(.dbd.O)R.sup.13;
or R.sup.7 and R.sup.8, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0457] R.sup.9 and
R.sup.10 are each independently selected from hydrogen, halogen,
alkyl, fluoroalkyl, --OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl;
or R.sup.9 and R.sup.10 together form an oxo; or optionally,
R.sup.9 and R.sup.1 together form a direct bond to provide a double
bond; or optionally, R.sup.9 and R.sup.1 together form a direct
bond, and R.sup.10 and R.sup.2 together form a direct bond to
provide a triple bond; [0458] R.sup.11 and R.sup.12 are each
independently selected from hydrogen, alkyl, carbocyclyl or
--C(.dbd.O)R.sup.23; or R.sup.11 and R.sup.12, together with the
nitrogen atom to which they are attached, form an N-heterocyclyl;
and [0459] each R.sup.13, R.sup.22 and R.sup.23 is independently
selected from alkyl, alkenyl, aryl, aralkyl, carbocyclyl,
heteroaryl or heterocyclyl; [0460] R.sup.6, R.sup.19, and R.sup.34
are each independently hydrogen or alkyl; [0461] each R.sup.33 is
independently selected from halogen, OR.sup.34, alkyl, or
fluoroalkyl; and n is 0, 1, 2, 3, or 4; [0462] R.sup.20 and
R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, --C(.dbd.O)R.sup.22; or R.sup.20 and R.sup.21,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; and [0463] each R.sup.24, R.sup.25, R.sup.26,
R.sup.27, R.sup.28 and R.sup.29 is independently selected from
hydrogen, alkyl, alkenyl, fluoroalkyl, aryl, heteroaryl,
carbocyclyl or heterocyclyl.
[0464] In another embodiment is the compound of Formula (Ia)
wherein, [0465] Z is --C(R.sup.9)(R.sup.10)--C)(R.sup.1)R.sup.2)--
or --O--C(R.sup.31)(R.sup.32)--; [0466] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0467] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0468] R.sup.3 and R.sup.4 are each independently
selected from hydrogen or alkyl; or R.sup.3 and R.sup.4 together
form an imino; [0469] R.sup.7 and R.sup.8 are each independently
selected from hydrogen, alkyl, carbocyclyl or --C(.dbd.O)R.sup.13;
or R.sup.7 and R.sup.8, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0470] R.sup.9 and
R.sup.10 are each independently selected from hydrogen, halogen,
alkyl, fluoroalkyl, --OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl;
or R.sup.9 and R.sup.10 together form an oxo; [0471] R.sup.11 and
R.sup.12 are each independently selected from hydrogen, alkyl,
carbocyclyl or --C(.dbd.O)R.sup.23; or R.sup.11 and R.sup.12,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; and [0472] each R.sup.13, R.sup.22 and R.sup.23 is
independently selected from alkyl, alkenyl, aryl, aralkyl,
carbocyclyl, heteroaryl or heterocyclyl; [0473] R.sup.6, R.sup.19,
and R.sup.34 are each independently hydrogen or alkyl; [0474] each
R.sup.33 is independently selected from halogen, OR.sup.34, alkyl,
or fluoroalkyl; and n is 0, 1, 2, 3, or 4; [0475] R.sup.20 and
R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, --C(.dbd.O)R.sup.22; or R.sup.20 and R.sup.21,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; and [0476] each R.sup.24, R.sup.25, R.sup.26,
R.sup.27, R.sup.28 and R.sup.29 is independently selected from
hydrogen, alkyl, alkenyl, fluoroalkyl, aryl, heteroaryl,
carbocyclyl or heterocyclyl.
[0477] In another embodiment is the compound of Formula (Ia)
wherein, G is selected from --N(R.sup.42)--SO.sub.2--R.sup.40;
R.sup.40 is selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl,
or heteroaryl.
[0478] In another embodiment is the compound of Formula (Ia) having
the structure of Formula (Ib)
##STR00039##
wherein, [0479] Z is --C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--
or --O--C(R.sup.31)(R.sup.32)--; [0480] R.sup.40 is selected from
--C(R.sup.16)(R.sup.17)(R.sup.18); [0481] R.sup.16 and R.sup.17 are
each independently selected from hydrogen, alkyl, halo, aryl,
heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or R.sup.16 and
R.sup.17, together with the carbon to which they are attached form
a carbocyclyl or heterocycle; [0482] R.sup.18 is selected from
hydrogen, alkyl, alkoxy, hydroxy, halo or fluoroalkyl; [0483]
R.sup.1 and R.sup.2 are each independently selected from hydrogen,
halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6 or
--NR.sup.7R.sup.8; or R.sup.1 and R.sup.2 together form an oxo;
[0484] R.sup.31 and R.sup.32 are each independently selected from
hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl; [0485] R.sup.3 and
R.sup.4 are each independently selected from hydrogen or alkyl; or
R.sup.3 and R.sup.4 together form an imino; [0486] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl or --C(.dbd.O)R.sup.13; or R.sup.7 and R.sup.8,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; [0487] R.sup.9 and R.sup.10 are each independently
selected from hydrogen, halogen, alkyl, fluoroalkyl, --OR.sup.19,
NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and R.sup.10 together
form an oxo; or optionally, R.sup.9 and R.sup.1 together form a
direct bond to provide a double bond; or optionally, R.sup.9 and
R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0488]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl or --C(.dbd.O)R.sup.23; or R.sup.11
and R.sup.12, together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0489] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or heterocyclyl;
[0490] R.sup.6, R.sup.19, and R.sup.34 are each independently
hydrogen or alkyl; [0491] each R.sup.33 is independently selected
from halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2,
3, or 4; [0492] R.sup.20 and R.sup.21 are each independently
selected from hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.22; or
[0493] R.sup.20 and R.sup.21, together with the nitrogen atom to
which they are attached, form an N-heterocyclyl; and [0494] each
R.sup.24, R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl.
[0495] In another embodiment is the compound of Formula (Ib)
wherein, R.sup.9 and R.sup.10 are each independently selected from
hydrogen, halogen, alkyl, fluoroalkyl, --OR.sup.19,
--NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and R.sup.10
together form an oxo.
[0496] In another embodiment is the compound of Formula (Ib) having
the structure of Formula (Ic):
##STR00040##
wherein, [0497] R.sup.1 and R.sup.2 are each independently selected
from hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl,
--OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and R.sup.2 together
form an oxo; [0498] R.sup.3 and R.sup.4 are each independently
selected from hydrogen or alkyl; or R.sup.3 and R.sup.4 together
form an imino; [0499] R.sup.7 and R.sup.8 are each independently
selected from hydrogen, alkyl, carbocyclyl or --C(.dbd.O)R.sup.13;
or R.sup.7 and R.sup.8, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0500] R.sup.9 and
R.sup.10 are each independently selected from hydrogen, halogen,
alkyl, fluoroalkyl, --OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl;
or R.sup.9 and R.sup.10 together form an oxo; [0501] R.sup.11 and
R.sup.12 are each independently selected from hydrogen, alkyl,
carbocyclyl or --C(.dbd.O)R.sup.23; or R.sup.11 and R.sup.12,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; [0502] each R.sup.13, R.sup.22 and R.sup.23 is
independently selected from alkyl, alkenyl, aryl, aralkyl,
carbocyclyl, heteroaryl or heterocyclyl; [0503] R.sup.6, R.sup.19
and R.sup.34 are each independently hydrogen or alkyl; [0504]
R.sup.20 and R.sup.21 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.22; or R.sup.20 and
R.sup.21, together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0505] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0506] R.sup.16 and
R.sup.17 are each independently selected from hydrogen,
C.sub.1-C.sub.13 alkyl, halo or fluoroalkyl; or R.sup.16 and
R.sup.17, together with the carbon to which they are attached form
a carbocyclyl or heterocycle; [0507] each R.sup.33 is independently
selected from halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is
0, 1, 2, 3, or 4; and R.sup.is is selected from a hydrogen, alkyl,
alkoxy, hydroxy, halo or fluoroalkyl.
[0508] In another embodiment is the compound of Formula (Ic)
wherein n is 0 and each of R.sup.11 and R.sup.12 is hydrogen. In a
further embodiment is the compound wherein each of R.sup.3,
R.sup.4, R.sup.14 and R.sup.15 is hydrogen. In a further embodiment
is the compound wherein, [0509] R.sup.1 and R.sup.2 are each
independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, or --OR.sup.6; [0510] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, or
--OR.sup.19; or R.sup.9 and R.sup.10 together form an oxo; [0511]
R.sup.6 and R.sup.19 are each independently hydrogen or alkyl;
[0512] R.sup.16 and R.sup.17, together with the carbon to which
they are attached form a carbocyclyl or heterocycle; and [0513]
R.sup.18 is selected from a hydrogen, alkoxy or hydroxy.
[0514] In a further embodiment is the compound wherein R.sup.16 and
R.sup.17, together with the carbon to which they are attached, form
a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or
cyclooctyl, and R.sup.18 is hydrogen or hydroxy.
[0515] In another embodiment is the compound of Formula (Ic),
wherein R.sup.11 is hydrogen and R.sup.12 is --C(.dbd.O)R.sup.23,
wherein R.sup.23 is alkyl. In further embodiment is the compound
wherein, R.sup.1 and R.sup.2 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, or --OR.sup.6; R.sup.9
and R.sup.10 are each independently selected from hydrogen,
halogen, alkyl, or --OR.sup.19; or R.sup.9 and R.sup.10 together
form an oxo; [0516] R.sup.6 and R.sup.19 are each independently
selected from hydrogen or alkyl; [0517] R.sup.16 and R.sup.17,
together with the carbon atom to which they are attached, form a
carbocyclyl; and [0518] R.sup.18 is hydrogen, hydroxy or
alkoxy.
[0519] In a further embodiment is the compound wherein [0520] n is
0; [0521] R.sup.16 and R.sup.17, together with the carbon atom to
which they are attached, form a cyclopentyl, cyclohexyl or
cyclohexyl; and [0522] R.sup.18 is hydrogen or hydroxy.
[0523] In a further embodiment is the compound of Formula (Ic),
wherein [0524] R.sup.1 and R.sup.2 are each independently selected
from hydrogen, halogen, C.sub.1-C.sub.5 alkyl or --OR.sup.6; [0525]
R.sup.9 and R.sup.10 are each independently selected from hydrogen,
halogen, alkyl, or --OR.sup.19; or R.sup.9 and R.sup.10 together
form an oxo; [0526] R.sup.6 and R.sup.19 are each independently
hydrogen or alkyl; [0527] R.sup.16 and R.sup.17 is independently
selected from C.sub.1-C.sub.13 alkyl; and R.sup.18 is hydrogen,
hydroxy or alkoxy.
[0528] In another embodiment is the compound of Formula (Ib) having
the structure of Formula (Id):
##STR00041##
wherein, [0529] R.sup.31 and R.sup.32 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0530] R.sup.3 and R.sup.4 are each independently selected from
hydrogen or alkyl; or R.sup.3 and R.sup.4 together form an imino;
[0531] R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, or --C(.dbd.O)R.sup.23; or R.sup.11
and R.sup.12, together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; [0532] R.sup.23 is selected from
alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;
[0533] R.sup.14 and R.sup.15 are each independently selected from
hydrogen or alkyl; [0534] R.sup.16 and R.sup.17 are each
independently selected from hydrogen, C.sub.1-C.sub.13 alkyl, halo
or fluoroalkyl; or R.sup.16 and R.sup.17, together with the carbon
atom to which they are attached, form a carbocyclyl or heterocycle;
[0535] R.sup.18 is selected from a hydrogen, alkyl, alkoxy,
hydroxy, halo or fluoroalkyl; [0536] R.sup.34 is hydrogen or alkyl;
and [0537] each R.sup.33 is independently selected from halogen,
OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or 4.
[0538] In another embodiment is the compound wherein n is 0 and
each of R.sup.11 and R.sup.12 is hydrogen. In another embodiment is
the compound wherein each R.sup.3, R.sup.4, R.sup.14 and R.sup.15
is hydrogen. In another embodiment is the compound wherein,
R.sup.31 and R.sup.32 are each independently hydrogen, or
C.sub.1-C.sub.5 alkyl; R.sup.16 and R.sup.17, together with the
carbon atom to which they are attached, form a carbocyclyl; and
R.sup.18 is hydrogen, hydroxy, or alkoxy. In a further embodiment
is the compound wherein R.sup.16 and R.sup.17, together with the
carbon atom to which they are attached form a cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl and
R.sup.18 is hydrogen or hydroxy. In a further embodiment is the
compound wherein, R.sup.31 and R.sup.32 are each independently
selected from hydrogen, or C.sub.1-C.sub.5 alkyl; and R.sup.18 is
hydrogen, hydroxy or alkoxy. In a further embodiment is the
compound wherein, R.sup.31 and R.sup.32 are each independently
hydrogen, or C.sub.1-C.sub.5 alkyl; R.sup.6 and R.sup.19 are each
independently hydrogen or alkyl; R.sup.16 and R.sup.17 is
independently selected from C.sub.1-C.sub.13 alkyl; and R.sup.18 is
hydrogen, hydroxy or alkoxy.
[0539] In another embodiment is the compound of Formula (I)
wherein, Z is a bond, --X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; and X is --S--,
--S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--, --C(.dbd.O)--,
--C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--. In a further embodiment is the compound
wherein, G is selected from --N(R.sup.42)--SO.sub.2--R.sup.40; and
R.sup.40 is selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl,
or heteroaryl.
[0540] In an additional embodiment is the compound of Formula (I)
having the structure of Formula (Ie):
##STR00042##
wherein, [0541] X is --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--,
--N(R.sup.30)--, --C(.dbd.O)--, --C(.dbd.CH.sub.2)--,
--C(.dbd.N--NR.sup.35)--, or --C(.dbd.N--OR.sup.35)--; [0542]
R.sup.31 and R.sup.32 are each independently selected from
hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl; [0543] R.sup.3 and
R.sup.4 are each independently selected from hydrogen or alkyl; or
R.sup.3 and R.sup.4 together form an imino; [0544] R.sup.11 and
R.sup.12 are each independently selected from hydrogen, alkyl,
carbocyclyl, or --C(.dbd.O)R.sup.23; or R.sup.11 and R.sup.12,
together with the nitrogen atom to which they are attached, form an
N-heterocyclyl; [0545] R.sup.23 is selected from alkyl, alkenyl,
aryl, carbocyclyl, heteroaryl or heterocyclyl; [0546] R.sup.16 and
R.sup.17 are each independently selected from hydrogen,
C.sub.1-C.sub.13 alkyl, halo or fluoroalkyl; or R.sup.16 and
R.sup.17, together with the carbon atom to which they are attached,
form a carbocyclyl or heterocycle; [0547] R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0548] R.sup.18
is selected from a hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; each R.sup.33 is independently selected from halogen,
OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or 4.
[0549] In a further embodiment is the compound of Formula (Ie)
wherein n is 0 and each R.sup.11 and R.sup.12 is hydrogen. In a
further embodiment is the compound wherein each R.sup.3, R.sup.4,
R.sup.14 and R.sup.15 is hydrogen. In a further embodiment is the
compound wherein, R.sup.31 and R.sup.32 are each independently
hydrogen, or C.sub.1-C.sub.5 alkyl; R.sup.16 and R.sup.17, together
with the carbon atom to which they are attached, form a carbocyclyl
or heterocycle; and R.sup.18 is hydrogen, hydroxy, or alkoxy. In a
further embodiment is the compound wherein R.sup.16 and R.sup.17,
together with the carbon atom to which they are attached form a
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or
cyclooctyl and R.sup.18 is hydrogen or hydroxy. In a further
embodiment is the compound wherein, R.sup.31 and R.sup.32 are each
independently selected from hydrogen, or C.sub.1-C.sub.5 alkyl;
R.sup.16 and R.sup.17 is independently selected from
C.sub.1-C.sub.13 alkyl; and R.sup.18 is hydrogen, hydroxy or
alkoxy.
[0550] In an additional embodiment is the compound of Formula (Ia)
wherein, G is selected from --N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40; R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.8), aryl, or
heteroaryl; each R.sup.42 is independently selected from hydrogen
or alkyl. In a further embodiment is the compound wherein, G is
selected from --N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40; R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; each R.sup.42 is independently selected from hydrogen
or alkyl. In a further embodiment is the compound wherein, R.sup.42
is a hydrogen; R.sup.40 is selected from
--C(R.sup.16)(R.sup.17)(R.sup.18); R.sup.16 and R.sup.17, together
with the carbon atom to which they are attached, form a carbocyclyl
or heterocycle; and R.sup.18 is hydrogen, hydroxy, or alkoxy. In a
further embodiment is the compound wherein, R.sup.42 is a hydrogen;
R.sup.40 is selected from --C(R.sup.16)(R.sup.17)(R.sup.18);
R.sup.16 and R.sup.17, together with the carbon atom to which they
are attached, form a carbocyclyl or heterocycle; and R.sup.18 is
hydrogen, hydroxy, or alkoxy.
[0551] In another embodiment is the compound of Formula (Ia)
wherein, [0552] G is selected from
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0553] each
R.sup.43 is independently selected from hydrogen, alkyl,
cycloalkyl, aralkyl, C-attached heterocyclyl, aryl, or heteroaryl;
or two R.sup.43 groups, together with the nitrogen to which they
are attached, may form a heterocyclyl; [0554] each R.sup.42 is
independently selected from hydrogen or alkyl.
[0555] In another embodiment is the compound Formula (Ia) wherein,
[0556] G is selected from
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0557] each
R.sup.43 is independently selected from hydrogen, alkyl,
cycloalkyl, aralkyl, C-attached heterocyclyl, aryl, or heteroaryl;
or two R.sup.43 groups, together with the nitrogen to which they
are attached, may form a heterocyclyl; and [0558] R.sup.42 is
hydrogen.
[0559] In a further embodiment is the compound wherein, [0560] each
R.sup.43 is independently selected from hydrogen, alkyl,
cycloalkyl, aralkyl, C-attached heterocyclyl, aryl, or heteroaryl;
or two R.sup.43 groups, together with the nitrogen to which they
are attached, may form a heterocyclyl; and [0561] R.sup.42 is
hydrogen.
[0562] In a further embodiment is the compound wherein, [0563]
R.sup.40 is selected from --C(R.sup.16)(R.sup.17)(R.sup.18); [0564]
R.sup.16 and R.sup.17 are each independently selected from
hydrogen, alkyl, halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or
fluoroalkyl; or R.sup.16 and R.sup.17, together with the carbon to
which they are attached form a carbocyclyl or heterocycle; [0565]
R.sup.18 is selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl.
[0566] In another embodiment is the compound of Formula (I) wherein
one, more than one, or all of the non-exchangeable .sup.1H atoms
have been substituted with .sup.2H atoms.
[0567] In a specific embodiment, the compound of Formula (I) is
selected from the group consisting of:
##STR00043## ##STR00044## ##STR00045## ##STR00046## ##STR00047##
##STR00048## ##STR00049## ##STR00050## ##STR00051## ##STR00052##
##STR00053## ##STR00054##
[0568] Additional Compounds of the Invention
[0569] In one embodiment is a compound having a structure of
Formula (II):
##STR00055##
as an isolated E or Z geometric isomer or a mixture of E and Z
geometric isomers, as a tautomer or a mixture of tautomers, as a
stereoisomer or as a pharmaceutically acceptable salt, hydrate,
solvate, N-oxide or prodrug thereof, wherein: R.sup.1 and R.sup.2
are each the same or different and independently hydrogen or alkyl;
R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are each the same or
different and independently hydrogen, halogen, nitro, --NH.sub.2,
--NHR.sup.13, --N(R.sup.13).sub.2, --OR.sup.12, alkyl or
fluoroalkyl; R.sup.7 and R.sup.8 are each the same or different and
independently hydrogen or alkyl; or R.sup.7 and R.sup.8 together
with the carbon atom to which they are attached, form a carbocyclyl
or heterocyclyl; or R.sup.7 and R.sup.8 together form an imino;
R.sup.9 is hydrogen, alkyl, carbocyclyl, heterocyclyl,
--C(.dbd.O)R.sup.13, --SO.sub.2R.sup.13, --CO.sub.2R.sup.13,
--CONH.sub.2, --CON(R.sup.13).sub.2 or --CON(H)R.sup.13; R.sup.10
is hydrogen or alkyl; or R.sup.9 and R.sup.10, together with the
nitrogen atom to which they are attached, form an N-heterocyclyl;
R.sup.11 is alkyl, alkenyl, aryl, carbocyclyl, heteroaryl or
heterocyclyl; each R.sup.12 is independently selected from hydrogen
or alkyl; each R.sup.13 is independently selected from alkyl,
carbocyclyl, heterocyclyl, aryl or heteroaryl; Z is a bond, Y or
W--Y, wherein W is --C(R.sup.14)(R.sup.15)--, --O--, --S--,
--S(.dbd.O)--, --S(.dbd.O).sub.2-- or --N(R.sup.12)_; Y is
--C(R.sup.16)(R.sup.17)-- or
--C(R.sup.16)(R.sup.7)--C(R.sup.21)(R.sup.22)--; R.sup.14 and
R.sup.15 are each the same or different and independently hydrogen,
halogen, alkyl, fluoroalkyl, --OR.sup.12, --NR.sup.18R.sup.19,
carbocyclyl or heterocyclyl; or R.sup.14 and R.sup.15 together form
an oxo, an imino, an oximo, or a hydrazino; R.sup.16 and R.sup.17
are each the same or different and independently hydrogen, halogen,
alkyl, fluoroalkyl, --OR.sup.12, --NR.sup.18R.sup.19, carbocyclyl
or heterocyclyl; or R.sup.16 and R.sup.17 together form an oxo; or
optionally, R.sup.14 and R.sup.16 together form a direct bond to
provide a double bond connecting W and Y; or optionally, R.sup.14
and R.sup.16 together form a direct bond, and R.sup.15 and R.sup.17
together form a direct bond to provide a triple bond connecting W
and Y; each R.sup.18 and R.sup.19 is independently selected from
hydrogen, alkyl, carbocyclyl, or --C(.dbd.O)R.sup.13,
--SO.sub.2R.sup.3, --CO.sub.2R.sup.13, --CONH.sub.2,
--CON(R.sup.13).sub.2 or --CON(H)R.sup.13; or R.sup.18 and
R.sup.19, together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; R.sup.21 and R.sup.22 are each
the same or different and independently hydrogen, halogen, alkyl,
fluoroalkyl, --OR.sup.12, --NR.sup.18R.sup.19, carbocyclyl or
heterocyclyl; provided that when R.sup.11 is phenyl, the compound
of Formula (A) is not: [0570]
2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]ace-
tamide; [0571]
(2S,3R)-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)-e-
thenyl]phenyl]-butanamide; [0572] L-glutamic acid,
1-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]ester;
glycine, 3-hydroxy-5-[(1E)-2-(4-hydroxyphenyl)ethenyl]phenyl ester;
[0573]
(2S)-2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)etheny-
l]phenyl]propanamide; [0574]
(2S)-2-amino-3-hydroxy-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)eth-
enyl]phenyl]propanamide; [0575] (2S)-2-amino-N-[2-methoxy-5-[(1
Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-4-methyl-pentanamide;
[0576] (2S)-2-amino-N-[2-methoxy-5-[(1
Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]-3-methyl-butanamide;
or [0577]
2-amino-N-[2-methoxy-5-[(1Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phe-
nylbutanamide; and wherein the compound of Formula (II) is
isotopically enriched.
[0578] In another embodiment is the compound of Formula (II) having
one, more than one, or all of the non exchangeable .sup.1H atoms
are replaced with .sup.2H atoms.
[0579] In another embodiment is the compound of Formula (II) having
the structure of Formula (IIa):
##STR00056##
wherein R.sup.11 is selected from:
##STR00057##
and one, more than one, or all of the non-exchangeable .sup.1H
atoms are replaced with .sup.2H atoms.
[0580] In another embodiment is the compound of Formula (IIa)
selected from:
##STR00058##
[0581] One embodiment provides a compound having a structure of
Formula (III):
##STR00059## [0582] as a tautomer or a mixture of tautomers, or as
a pharmaceutically acceptable salt, hydrate, solvate, N-oxide,
stereoisomer, geometric isomer or prodrug thereof, wherein: [0583]
m is 0, 1, 2 or 3; [0584] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.21)(R.sup.22)--,
--C(R.sup.23)(R.sup.24)--C(R.sup.1)(R.sup.2)--, or
--C(R.sup.23)(R.sup.24)--C(R.sup.25)(R.sup.26)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.21)(R.sup.22)--C(R.sup.1)(R.sup.2)--,
--C(R.sup.32)(R.sup.33)--X--C(R.sup.21)(R.sup.22)--; [0585] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.31)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0586] Y is a bond,
--C(R.sup.27)(R.sup.28)--, or
--C(R.sup.27)(R.sup.2)--C(R.sup.29)(R.sup.30)--; [0587] R.sup.1 and
R.sup.2 are each independently selected from hydrogen, halogen,
C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6 or
--NR.sup.7R.sup.8; or R.sup.1 and R.sup.2 together form an oxo;
[0588] R.sup.21, R.sup.22, R.sup.32 and R.sup.33 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0589] R.sup.23 and R.sup.24 are each independently
selected from hydrogen, halogen, C.sub.1-C.sub.5 alkyl,
fluoroalkyl, --OR.sup.6, --NR.sup.7R.sup.8; or R.sup.23 and
R.sup.24 together form an oxo; or optionally, R.sup.23 and an
adjacent R.sup.1 together form a direct bond to provide a double
bond; or optionally, R.sup.23 and an adjacent R.sup.1 together form
a direct bond, and R.sup.24 and an adjacent R.sup.2 together form a
direct bond to provide a triple bond; [0590] R.sup.25 and R.sup.26
are each independently selected from hydrogen, halogen,
C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6 or
--NR.sup.7R.sup.8; or R.sup.25 and R.sup.26 together form an oxo;
[0591] R.sup.3 and R.sup.4 are each independently selected from
hydrogen, alkyl, heteroalkyl, alkenyl, fluoroalkyl, aryl,
heteroaryl, carbocyclyl or C-attached heterocyclyl; or R.sup.3 and
R.sup.4 together with the carbon atom to which they are attached,
form a carbocyclyl or heterocyclyl; or R.sup.3 and R.sup.4 together
form an imino; [0592] R.sup.5 is alkyl, heteroalkyl, alkenyl,
heteroalkenyl, aryl, carbocyclyl, heteroaryl or heterocyclyl;
[0593] each R.sup.6 is the same or different and independently
hydrogen or C.sub.1-C.sub.5 alkyl; [0594] each R.sup.7 and each
R.sup.8 are each the same or different and independently hydrogen,
alkyl, carbocyclyl, heteroalkyl, heterocycloalkyl, aryl,
heteroaryl, --C(.dbd.O)R.sup.9, SO.sub.2R.sup.9, CO.sub.2R.sup.9,
SO.sub.2NH.sub.2, SO.sub.2NHR.sup.9 or SO.sub.2N(R.sup.9).sub.2; or
R.sup.7 and R.sup.8, together with the nitrogen atom to which they
are attached, form an N-heterocyclyl; [0595] each R.sup.9 is the
same or different and each is independently alkyl, alkenyl, aryl,
carbocyclyl, heteroaryl or heterocyclyl; [0596] R.sup.12 and
R.sup.13 are the same or different and independently hydrogen,
alkyl, heteroalkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl,
--C(.dbd.O)R.sup.9, SO.sub.2R.sup.9, CO.sub.2R.sup.9,
SO.sub.2NH.sub.2, SO.sub.2NHR.sup.9 or SO.sub.2N(R.sup.9).sub.2; or
R.sup.12 and R.sup.13 together with the nitrogen atom to which they
are attached, form an N-heterocyclyl; and [0597] each R.sup.14 is
the same or different and independently alkyl, halo, fluoroalkyl or
--OR.sup.6; [0598] each R.sup.27, R.sup.28, R.sup.29 and R.sup.31
are the same or different and independently hydrogen, alkyl or
--OR.sup.6; and [0599] R.sup.30 and R.sup.35 are each independently
hydrogen or C.sub.1-C.sub.5 alkyl; and wherein [0600] the compound
of Formula (III) is isotopically enriched.
[0601] Another embodiment provides the compound of Formula (III)
having one, more than one or all of the non-exchangeable .sup.1H
atoms replaced with .sup.2H atoms.
[0602] Another embodiment provides the compound of Formula
(IIIa):
##STR00060##
wherein Y is a bond; [0603] R.sup.5 is selected from:
##STR00061##
[0603] and one, more than one, or all of the non-exchangeable
.sup.1H atoms are replaced with .sup.2H atoms.
[0604] Another embodiment provides the compound of Formula (III)
selected from:
##STR00062##
[0605] One embodiment provides a compound of Formula (IV) or
tautomer, stereoisomer, geometric isomer or a pharmaceutically
acceptable solvate, hydrate, salt, N-oxide or prodrug thereof:
##STR00063##
wherein, [0606] Z is --C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--
or --X--C(R.sup.3)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0607] R.sup.1
and R.sup.2 are each independently selected from hydrogen, halogen,
C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6 or
--NR.sup.7R.sup.8; or R.sup.1 and R.sup.2 together form an oxo;
[0608] R.sup.31 and R.sup.32 are each independently selected from
hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl; [0609] R.sup.36
and R.sup.37 are each independently selected from hydrogen,
halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6 or
--NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an oxo;
or optionally, R.sup.36 and R.sup.1 together form a direct bond to
provide a double bond; or optionally, R.sup.36 and R.sup.1 together
form a direct bond, and R.sup.37 and R.sup.2 together form a direct
bond to provide a triple bond; [0610] R.sup.3 and R.sup.4 are each
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or C-attached heterocyclyl; or
R.sup.3 and R.sup.4 together with the carbon atom to which they are
attached, form a carbocyclyl or heterocyclyl; or R.sup.3 and
R.sup.4 together form an imino; [0611] R.sup.5 is C.sub.5-C.sub.15
alkyl or carbocyclyalkyl; [0612] R.sup.7 and R.sup.8 are each
independently selected from hydrogen, alkyl, carbocyclyl,
heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0613] X is --O--, --S--, --S(.dbd.O)--,
--S(.dbd.O).sub.2--, --N(R.sup.30)--, --C(.dbd.O)--,
--C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0614] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0615]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0616] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0617] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0618] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0619] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0620] each R.sup.33
is independently selected from halogen, OR.sup.34, alkyl, or
fluoroalkyl; and n is 0, 1, 2, 3, or 4; with the provision that
R.sup.5 is not 2-(cyclopropyl)-1-ethyl or an unsubstituted normal
alkyl; and wherein [0621] the compound of Formula (IV) is
isotopically enriched.
[0622] Another embodiment provides the compound of Formula (IV) has
one, more than one or all of the non-exchangeable .sup.1H atoms
replaced with .sup.2H atoms.
[0623] Another embodiment provides the compound having the
structure of Formula (IVa):
##STR00064##
wherein Y is a bond; [0624] R.sup.5 is selected from:
##STR00065##
[0624] and one, more than one, or all of the non-exchangeable
.sup.1H atoms are replaced with .sup.2H atoms.
[0625] Another embodiment provides the compound selected from:
##STR00066##
[0626] One embodiment provides a method for treating an ophthalmic
disease or disorder in a subject, comprising administering to the
subject a compound of Formula (I) or tautomer, stereoisomer,
geometric isomer or a pharmaceutically acceptable solvate, hydrate,
salt, N-oxide or prodrug thereof:
##STR00067##
wherein, [0627] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.3)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0628] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0629] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0630] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0631] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0632] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0633] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0634] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0635] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0636] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0637] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0638] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0639] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0640]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0641] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0642] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0643] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0644] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0645] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0646] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0647] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0648] Another embodiment provides a method for treating an
ophthalmic disease or disorder wherein the ophthalmic disease or
disorder is age-related macular degeneration or Stargardt's macular
dystrophy.
[0649] In a further embodiment is the method wherein the ophthalmic
disease or disorder is a retinal disease or disorder. In an
additional embodiment is the method wherein the retinal disease or
disorder is age-related macular degeneration or Stargardt's macular
dystrophy. In an additional embodiment is the method wherein the
ophthalmic disease or disorder is selected from retinal detachment,
hemorrhagic retinopathy, retinitis pigmentosa, optic neuropathy,
inflammatory retinal disease, proliferative vitreoretinopathy,
retinal dystrophy, hereditary optic neuropathy, Sorsby's fundus
dystrophy, uveitis, a retinal injury, a retinal disorder associated
with Alzheimer's disease, a retinal disorder associated with
multiple sclerosis, a retinal disorder associated with Parkinson's
disease, a retinal disorder associated with viral infection, a
retinal disorder related to light overexposure, and a retinal
disorder associated with AIDS. In an additional embodiment is the
method wherein the ophthalmic disease or disorder is selected from
diabetic retinopathy, diabetic maculopathy, retinal blood vessel
occlusion, retinopathy of prematurity, or ischemia reperfusion
related retinal injury.
[0650] One embodiment provides a method for treating an ophthalmic
disease or disorder in a subject, comprising administering to the
subject a compound of Formula (II), (IIa), (III), (IIIa), (IV), or
(IVa) as described herein, or tautomer, stereoisomer, geometric
isomer or a pharmaceutically acceptable solvate, hydrate, salt,
N-oxide or prodrug thereof. Another embodiment provides a method
for treating an ophthalmic disease or disorder wherein the
ophthalmic disease or disorder is age-related macular degeneration
or Stargardt's macular dystrophy.
[0651] In an additional embodiment is the method of inhibiting at
least one visual cycle trans-cis isomerase in a cell comprising
contacting the cell with a compound of Formula (I) as described
herein, thereby inhibiting the at least one visual cycle trans-cis
isomerase. In a further embodiment is the method wherein the cell
is a retinal pigment epithelial (RPE) cell.
[0652] In a further embodiment is the method of inhibiting at least
one visual cycle trans-cis isomerase in a subject comprising
administering to the subject the pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound of
Formula (I) or tautomer, stereoisomer, geometric isomer or a
pharmaceutically acceptable solvate, hydrate, salt, N-oxide or
prodrug thereof:
##STR00068##
wherein, [0653] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.1)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0654] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0655] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0656] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0657] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0658] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0659] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0660] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0661] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0662] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0663] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0664] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0665] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0666]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0667] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0668] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0669] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0670] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0671] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0672] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0673] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0674] In a further embodiment is the method wherein the subject is
human. In a further embodiment is the method wherein accumulation
of lipofuscin pigment is inhibited in an eye of the subject. In a
further embodiment is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E). In a further
embodiment is the method wherein degeneration of a retinal cell is
inhibited. In a further embodiment is the method wherein the
retinal cell is a retinal neuronal cell. In a further embodiment is
the method wherein the retinal neuronal coil is a photoreceptor
cell, an amacrine cell, a horizontal cell, a ganglion cell, or a
bipolar cell. In a further embodiment is the method wherein the
retinal cell is a retinal pigment epithelial (RPE) cell.
[0675] In an additional embodiment is a compound that inhibits
11-cis-retinol production with an IC.sub.50 of about 1 .mu.M or
less when assayed in vitro, utilizing extract of cells that express
RPE65 and LRAT, wherein the extract further comprises CRALBP,
wherein the compound is stable in solution for at least about 1
week at room temperature. In an additional embodiment, the compound
is a non-retinoid compound. In a further embodiment is the
compound, wherein the compound inhibits 11-cis-retinol production
with an IC.sub.50 of about 0.1 .mu.M or less. In a further
embodiment is the compound, wherein the compound inhibits
11-cis-retinol production with an IC.sub.50 of about 0.01 .mu.M or
less.
[0676] In an additional embodiment is a non-retinoid compound that
inhibits an 11-cis-retinol producing isomerase reaction, wherein
said isomerase reaction occurs in RPE, and wherein said compound
has an ED.sub.50 value of 1 mg/kg or less when administered to a
subject. In a further embodiment is the non-retinoid compound
wherein the ED.sub.50 value is measured after administering a
single dose of the compound to said subject for about 2 hours or
longer.
[0677] In a further embodiment is the non-retinoid compound wherein
the structure of the non-retinoid compound corresponds to Formula
(I) or tautomer, stereoisomer, geometric isomer or a
pharmaceutically acceptable solvate, hydrate, salt, N-oxide or
prodrug thereof:
##STR00069##
wherein, [0678] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0679] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0680] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0681] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0682] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0683] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0684] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0685] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0686] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0687] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0688] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0689] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0690] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0691]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0692] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0693] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0694] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0695] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0696] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0697] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0698] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0699] In an additional embodiment is a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound
that inhibits 11-cis-retinol production with an IC.sub.50 of about
1 .mu.M or less when assayed in vitro, utilizing extract of cells
that express RPE65 and LRAT, wherein the extract further comprises
CRALBP, wherein the compound is stable in solution for at least
about 1 week at room temperature. In an additional embodiment is a
pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a non-retinoid compound that inhibits an 11-cis-retinol
producing isomerase reaction, wherein said isomerase reaction
occurs in RPE, and wherein said compound has an ED.sub.50 value of
1 mg/kg or less when administered to a subject.
[0700] In an additional embodiment is a method of modulating
chromophore flux in a retinoid cycle comprising introducing into a
subject a compound of Formula (I) as described herein. In a further
embodiment is the method resulting in a reduction of lipofuscin
pigment accumulated in an eye of the subject. In another embodiment
is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E). In yet another
embodiment is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E).
[0701] In an additional embodiment is a method of modulating
chromophore flux in a retinoid cycle comprising introducing into a
subject a compound that inhibits 11-cis-retinol production as
described herein. In a further embodiment is the method resulting
in a reduction of lipofuscin pigment accumulated in an eye of the
subject. In another embodiment is the method wherein the lipofuscin
pigment is N-retinylidene-N-retinyl-ethanolamine (A2E). In yet
another embodiment is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E).
[0702] In an additional embodiment is a method of modulating
chromophore flux in a retinoid cycle comprising introducing into a
subject a non-retinoid compound that inhibits an 11-cis-retinol
producing isomerase reaction as described herein. In a further
embodiment is the method resulting in a reduction of lipofuscin
pigment accumulated in an eye of the subject. In another embodiment
is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E). In yet another
embodiment is the method wherein the lipofuscin pigment is
N-retinylidene-N-retinyl-ethanolamine (A2E).
[0703] In an additional embodiment is a method for treating an
ophthalmic disease or disorder in a subject, comprising
administering to the subject a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound
that inhibits 11-cis-retinol production with an IC.sub.50 of about
1 .mu.M or less when assayed in vitro, utilizing extract of cells
that express RPE65 and LRAT, wherein the extract further comprises
CRALBP, wherein the compound is stable in solution for at least
about 1 week at room temperature. In a further embodiment is the
method wherein the ophthalmic disease or disorder is age-related
macular degeneration or Stargardt's macular dystrophy. In a further
embodiment is the method wherein the ophthalmic disease or disorder
is selected from retinal detachment, hemorrhagic retinopathy,
retinitis pigmentosa, cone-rod dystrophy, Sorsby's fundus
dystrophy, optic neuropathy, inflammatory retinal disease, diabetic
retinopathy, diabetic maculopathy, retinal blood vessel occlusion,
retinopathy of prematurity, or ischemia reperfusion related retinal
injury, proliferative vitreoretinopathy, retinal dystrophy,
hereditary optic neuropathy, Sorsby's fundus dystrophy, uveitis, a
retinal injury, a retinal disorder associated with Alzheimer's
disease, a retinal disorder associated with multiple sclerosis, a
retinal disorder associated with Parkinson's disease, a retinal
disorder associated with viral infection, a retinal disorder
related to light overexposure, myopia, and a retinal disorder
associated with AIDS. In a further embodiment is the method
resulting in a reduction of lipofuscin pigment accumulated in an
eye of the subject.
[0704] In an additional embodiment is a method for treating an
ophthalmic disease or disorder in a subject, comprising
administering to the subject a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a non-retinoid
compound that inhibits an 11-cis-retinol producing isomerase
reaction, wherein said isomerase reaction occurs in RPE, and
wherein said compound has an ED.sub.50 value of 1 mg/kg or less
when administered to a subject. In a further embodiment is the
method wherein the ophthalmic disease or disorder is age-related
macular degeneration or Stargardt's macular dystrophy. In a further
embodiment is the method wherein the ophthalmic disease or disorder
is selected from retinal detachment, hemorrhagic retinopathy,
retinitis pigmentosa, cone-rod dystrophy, Sorsby's fundus
dystrophy, optic neuropathy, inflammatory retinal disease, diabetic
retinopathy, diabetic maculopathy, retinal blood vessel occlusion,
retinopathy of prematurity, or ischemia reperfusion related retinal
injury, proliferative vitreoretinopathy, retinal dystrophy,
hereditary optic neuropathy, Sorsby's fundus dystrophy, uveitis, a
retinal injury, a retinal disorder associated with Alzheimer's
disease, a retinal disorder associated with multiple sclerosis, a
retinal disorder associated with Parkinson's disease, a retinal
disorder associated with viral infection, a retinal disorder
related to light overexposure, myopia, and a retinal disorder
associated with AIDS. In a further embodiment is the method
resulting in a reduction of lipofuscin pigment accumulated in an
eye of the subject.
[0705] In a further embodiment is a method of inhibiting dark
adaptation of a rod photoreceptor cell of the retina comprising
contacting the retina with a compound of Formula (I) as described
herein.
[0706] In a further embodiment is a method of inhibiting dark
adaptation of a rod photoreceptor cell of the retina comprising
contacting the retina with a compound that inhibits 11-cis-retinol
production as described herein.
[0707] In a further embodiment is a method of inhibiting dark
adaptation of a rod photoreceptor cell of the retina comprising
contacting the retina with a non-retinoid compound that inhibits an
11-cis-retinol producing isomerase reaction as described
herein.
[0708] In a further embodiment is a method of inhibiting
regeneration of rhodopsin in a rod photoreceptor cell of the retina
comprising contacting the retina with a compound of Formula (I) as
described herein.
[0709] In a further embodiment is a method of inhibiting
regeneration of rhodopsin in a rod photoreceptor cell of the retina
comprising contacting the retina with a compound that inhibits
11-cis-retinol production as described herein.
[0710] In a further embodiment is a method of inhibiting
regeneration of rhodopsin in a rod photoreceptor cell of the retina
comprising contacting the retina with a non-retinoid compound that
inhibits an 11-cis-retinol producing isomerase reaction as
described herein.
[0711] In a further embodiment is a method of reducing ischemia in
an eye of a subject comprising administering to the subject the
pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a compound of Formula (I) or tautomer, stereoisomer,
geometric isomer or a pharmaceutically acceptable solvate, hydrate,
salt, N-oxide or prodrug thereof:
##STR00070##
wherein, [0712] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0713] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0714] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0715] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0716] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0717] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0718] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0719] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0720] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0721] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0722] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0723] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0724] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0725]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0726] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0727] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0728] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0729] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0730] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0731] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0732] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0733] In another embodiment is a method of reducing ischemia in an
eye of a subject comprising administering to the subject a
pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a compound that inhibits 11-cis-retinol production with
an IC.sub.50 of about 1 .mu.M or less when assayed in vitro,
utilizing extract of cells that express RPE65 and LRAT, wherein the
extract further comprises CRALBP, wherein the compound is stable in
solution for at least about 1 week at room temperature. In a
further embodiment is the method wherein the pharmaceutical
composition is administered under conditions and at a time
sufficient to inhibit dark adaptation of a rod photoreceptor cell,
thereby reducing ischemia in the eye.
[0734] In another embodiment is a method of reducing ischemia in an
eye of a subject comprising administering to the subject a
pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a non-retinoid compound that inhibits an 11-cis-retinol
producing isomerase reaction, wherein said isomerase reaction
occurs in RPE, and wherein said compound has an ED.sub.50 value of
1 mg/kg or less when administered to a subject. In a further
embodiment is the method wherein the pharmaceutical composition is
administered under conditions and at a time sufficient to inhibit
dark adaptation of a rod photoreceptor cell, thereby reducing
ischemia in the eye.
[0735] In another embodiment is a method of inhibiting
neovascularization in the retina of an eye of a subject comprising
administering to the subject a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound
that inhibits 11-cis-retinol production with an IC.sub.50 of about
1 .mu.M or less when assayed in vitro, utilizing extract of cells
that express RPE65 and LRAT, wherein the extract further comprises
CRALBP, wherein the compound is stable in solution for at least
about 1 week at room temperature. In a further embodiment is the
method wherein the pharmaceutical composition is administered under
conditions and at a time sufficient to inhibit dark adaptation of a
rod photoreceptor cell, thereby inhibiting neovascularization in
the retina.
[0736] In another embodiment is a method of inhibiting
neovascularization in the retina of an eye of a subject comprising
administering to the subject a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a non-retinoid
compound that inhibits an 11-cis-retinol producing isomerase
reaction, wherein said isomerase reaction occurs in RPE, and
wherein said compound has an ED.sub.50 value of 1 mg/kg or less
when administered to a subject. In a further embodiment is the
method wherein the pharmaceutical composition is administered under
conditions and at a time sufficient to inhibit dark adaptation of a
rod photoreceptor cell, thereby inhibiting neovascularization in
the retina.
[0737] In another embodiment is a method of inhibiting degeneration
of a retinal cell in a retina comprising contacting the retina with
the compound of Formula (I) as described herein. In a further
embodiment is the method wherein the retinal cell is a retinal
neuronal cell. In yet another embodiment is the method wherein the
retinal neuronal cell is a photoreceptor cell.
[0738] In another embodiment is a method of inhibiting degeneration
of a retinal cell in a retina comprising contacting the retina with
a compound that inhibits 11-cis-retinol production with an
IC.sub.50 of about 1 .mu.M or less when assayed in vitro, utilizing
extract of cells that express RPE65 and LRAT, wherein the extract
further comprises CRALBP, wherein the compound is stable in
solution for at least about 1 week at room temperature. In a
further embodiment is the method wherein the retinal cell is a
retinal neuronal cell. In yet another embodiment is the method
wherein the retinal neuronal cell is a photoreceptor cell.
[0739] In another embodiment is a method of inhibiting degeneration
of a retinal cell in a retina comprising contacting the retina with
a non-retinoid compound that inhibits an 11-cis-retinol producing
isomerase reaction, wherein said isomerase reaction occurs in RPE,
and wherein said compound has an ED.sub.50 value of 1 mg/kg or less
when administered to a subject. In a further embodiment is the
method wherein the retinal cell is a retinal neuronal cell. In yet
another embodiment is the method wherein the retinal neuronal cell
is a photoreceptor cell.
[0740] In another embodiment is a method of reducing lipofuscin
pigment accumulated in a subject's retina comprising administering
to the subject a pharmaceutical composition comprising a
pharmaceutically acceptable carrier and a compound of Formula (I)
or tautomer, stereoisomer, geometric isomer or a pharmaceutically
acceptable solvate, hydrate, salt, N-oxide or prodrug thereof:
##STR00071##
wherein, [0741] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.3)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [0742] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [0743] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [0744] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [0745] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [0746] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [0747] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [0748] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [0749] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
[0750] R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [0751] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [0752] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [0753] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [0754]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [0755] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [0756] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [0757] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [0758] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [0759] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [0760] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [0761] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[0762] In a further embodiment is the method wherein the lipofuscin
is N-retinylidene-N-retinyl-ethanolamine (A2E).
[0763] In another embodiment is a method of reducing lipofuscin
pigment accumulated in a subject's retina comprising administering
to the subject a pharmaceutical composition comprising a
pharmaceutically acceptable carrier and a compound that inhibits
11-cis-retinol production with an IC.sub.50 of about 1 .mu.M or
less when assayed in vitro, utilizing extract of cells that express
RPE65 and LRAT, wherein the extract further comprises CRALBP,
wherein the compound is stable in solution for at least about 1
week at room temperature. In a further embodiment is the method
wherein the lipofuscin is N-retinylidene-N-retinyl-ethanolamine
(A2E).
[0764] In another embodiment is a method of reducing lipofuscin
pigment accumulated in a subject's retina comprising administering
to the subject a pharmaceutical composition comprising a
pharmaceutically acceptable carrier and a non-retinoid compound
that inhibits an 11-cis-retinol producing isomerase reaction,
wherein said isomerase reaction occurs in RPE, and wherein said
compound has an ED.sub.50 value of 1 mg/kg or less when
administered to a subject. In a further embodiment is the method
wherein the lipofuscin is N-retinylidene-N-retinyl-ethanolamine
(A2E).
[0765] Certain compounds disclosed herein have the structures shown
in Table 1. The example number refers to a specific Example herein
that describes the preparation of the compound having the
structure/name shown.
TABLE-US-00001 TABLE 1 Ex- ample No. Structure Name 1 ##STR00072##
N-(3-(2-Aminoethoxy)phenyl) pentane-2-sulfonamide 2 ##STR00073##
N-(3-(2-Aminoethoxy)phenyl) butane-2-sulfonamide 3 ##STR00074##
N-(3-(2-Aminoethoxy)phenyl) propane-2-sulfonamide hydrochloride 4
##STR00075## N-(3-(2-Aminoethoxy) phenyl)cyclohexanesulfonamide
hydrochloride 5 ##STR00076## N-(3-(3-Amino-1- hydroxypropyl)phenyl)
cyclohexanesulfonamide 6 ##STR00077## N-(3-(3- Aminopropyl)phenyl)
cyclohexanesulfonamide 7 ##STR00078## 3-(3-Aminopropyl)-N-
(cyclohexylmethyl)aniline 8 ##STR00079## N-(3-(3-
Aminopropyl)phenyl) cyclohexanecarboxamide 9 ##STR00080##
3-(3-(3-Aminopropyl)phenyl)- 1,1-dipropylurea 10 ##STR00081##
1-(3-(2-Aminoethoxy) phenyl)-3- cyclohexylthiourea 11 ##STR00082##
3-Amino-1-(3- (cyclohexylmethylamino) phenyl)propan-1-ol 12
##STR00083## 3-Amino-1-(3- (cyclohexylmethylamino)
phenyl)propan-1-one 13 ##STR00084## 3-Amino-1-(3-
(pentylamino)phenyl) propan-1-ol 14 ##STR00085## 3-Amino-1-(3-
(pentylamino)phenyl) propan-1-one 15 ##STR00086## N-(3-(3-Amino-1-
hydroxypropyl)phenyl) cyclohexanecarboxamide 16 ##STR00087##
N-(3-(3- Aminopropanoyl)phenyl) cyclohexanecarboxamide 17
##STR00088## N-(3-(3-Amino-1- hydroxypropyl)phenyl) pentanamide 18
##STR00089## N-(3-(3- Aminopropanoyl)phenyl) pentanamide 19
##STR00090## 3-(3-Amino-1-fluoropropyl)-N-
(cyclohexylmethyl)aniline 20 ##STR00091## N-(3-(3-
Aminopropanoyl)phenyl) cyclohexanesulfonamide 21 ##STR00092##
N-(3-(3-Amino-1- hydroxypropyl)phenyl) butane-1- sulfonamide 22
##STR00093## N-(3-(3- Aminopropanoyl)phenyl) butane-1-sulfonamide
23 ##STR00094## (E)-3-(3-Aminoprop- 1-enyl)-N-
(cyclohexylmethyl)aniline 24 ##STR00095## 3-(3-Aminoprop-1-ynyl)-N-
(cyclohexylmethyl)aniline 25 ##STR00096## (E)-N-(3-(3-Aminoprop-1-
enyl)phenyl)cyclo- hexanecarboxamide 26 ##STR00097##
N-(3-(3-Aminoprop-1- ynyl)phenyl)cyclo- hexanecarboxamide 27
##STR00098## (E)-N-(3-(3-Aminoprop-1- enyl)phenyl)cyclo-
hexanesulfonamide 28 ##STR00099## N-(3-(3-Aminoprop-1-
ynyl)phenyl)cyclo- hexanesulfonamide 29 ##STR00100##
(E)-1-((3-(3-Aminoprop-1- enyl)phenylamino)methyl) cyclohexanol 30
##STR00101## 1-((3-(3-Aminoprop-1- ynyl)phenylamino)methyl)
cyclohexanol 31 ##STR00102## 3-(3-Aminopropyl)-N-
(cyclopentylmethyl)aniline 32 ##STR00103## 3-(3-Aminopropyl)-N-(2-
propylpentyl)aniline 33 ##STR00104## 3-(3-Aminopropyl)-N-(2-
ethylbutyl)aniline 34 ##STR00105## 3-(3-Aminopropyl)-N-
benzylaniline 35 ##STR00106## 3-Amino-1-(3-(2-
ethylbutylamino)phenyl) propan-1-ol 36 ##STR00107##
3-Amino-1-(3-(2- ethylbutylamino)phenyl) propan-1-one 37
##STR00108## 3-Amino-1-(3-(2- propylpentylamino)phenyl) propan-1-ol
38 ##STR00109## 3-Amino-1-(3-(2- propylpentylamino)phenyl)
propan-1-one 39 ##STR00110## 3-Amino-1-(3- (cyclopentylmethylamino)
phenyl)propan-1-ol 40 ##STR00111## 3-Amino-1-(3-
(cyclopentylmethylamino) phenyl)propan-1-one 41 ##STR00112##
3-Amino-1-(3-(5- (benzyloxy)pentylamino) phenyl)propan-1-ol 42
##STR00113## 3-Amino-1-(3-(5- (benzyloxy)pentylamino)
phenyl)propan-1-one 43 ##STR00114## 5-(3-(3-Amino-1-
hydroxypropyl)phenylamino) pentan-1-ol 44 ##STR00115##
3-Amino-1-(3-(5- hydroxypentylamino)phenyl) propan-1-one 45
##STR00116## 3-Amino-1-(3-(5- methoxypentylamino)
phenyl)propan-1-ol 46 ##STR00117## 3-Amino-1-(3-(5-
methoxypentylamino)phenyl) propan-1-one 47 ##STR00118##
3-Amino-1-(3-((2- methoxybenzyl)amino)phenyl) propan-1-ol 48
##STR00119## 3-Amino-1-(3-(2- methoxybenzylamino)
phenyl)propan-1-one 49 ##STR00120## 3-Amino-1-(3-
(phenethylamino)phenyl) propan-1-ol 50 ##STR00121## 3-Amino-1-(3-
(phenethylamino)phenyl) propan-1-one 51 ##STR00122##
3-Amino-1-(3-(3- cyclohexylpropylamino) phenyl)propan-1-ol 52
##STR00123## 3-Amino-1-(3-(3- cyclohexylpropylamino)
phenyl)propan-1-one 53 ##STR00124## 4-((3-(3-Amino-1-
hydroxypropyl)phenylamino) methyl)heptan-4-ol 54 ##STR00125##
3-Amino-1-(3-(2-hydroxy-2- propylpentylamino) phenyl)propan-1-one
55 ##STR00126## 1-((3-(3-Amino-1- hydroxypropyl)phenylamino)
methyl)cyclohexanol 56 ##STR00127## 3-Amino-1-(3-((1-
hydroxycyclohexyl)methylamino) phenyl)propan-1-one 57 ##STR00128##
N-(3-(3-amino-2,2- dideutero-1- hydroxypropyl)phenyl)
cyclohexanecarboxamide 58 ##STR00129## N-(3-(3-amino-2,2-
dideutero-1- hydroxypropyl)phenyl) cyclohexanesulfonamide 59
##STR00130## 3-Amino-1-(3-(3- phenylpropylamino) phenyl)propan-1-ol
60 ##STR00131## 3-Amino-1-(3-(3- phenylpropylamino)
phenyl)propan-1-one 61 ##STR00132## 3-Amino-1-(3-((4,4-
difluorocyclohexyl) methylamino)phenyl) propan-1-ol 62 ##STR00133##
3-Amino-1-(3-((4,4- difluorocyclohexyl) methylamino)phenyl)
propan-1-one 63 ##STR00134## 3-(3-Aminopropyl)-N-((4,4-
difluorocyclohexyl) methyl)aniline 64 ##STR00135##
3-(3-Aminopropyl)-N-(3- phenylpropyl)aniline 65 ##STR00136##
3-(3-Aminopropyl)-N-(5- methoxypentyl)aniline 66 ##STR00137##
5-(3-(3- Aminopropyl)phenylamino) pentan-1-ol 67 ##STR00138##
4-((3-(3- Aminopropyl)phenylamino) methyl)heptan-4-ol 68
##STR00139## 3-((3-(3- Aminopropyl)phenylamino) methyl)pentan-3-ol
69 ##STR00140## 1-((3-3- Aminopropyl)phenylamino)
methyl)cyclohexanol 70 ##STR00141## 1-((3-(3-
Aminopropyl)phenylamino) methyl)cyclopentanol 71 ##STR00142##
N-(3-(3-Aminopropyl) phenyl)-2- propylpentanamide 72 ##STR00143##
N-(3-(3-Aminopropyl) phenyl)heptane- 4-sulfonamide 73 ##STR00144##
N-(3-(3-Amino-1- hydroxypropyl)phenyl)-2- propylpentanamide 74
##STR00145## N-(3-(3-Amino-1- hydroxypropyl)phenyl)
heptane-4-sulfonamide 75 ##STR00146## N-(3-(3-Aminopropanoyl)
phenyl)-2- propylpentanamide 76 ##STR00147## N-(3-(3-
Aminopropanoyl)phenyl) heptane-4-sulfonamide 77 ##STR00148##
3-((3-(3-Amino-1- hydroxypropyl)phenylamino) methyl)pentan-3-ol 78
##STR00149## 1-((3-(3-Amino-1- hydroxypropyl)phenylamino)
methyl)cyclopentanol 79 ##STR00150## 3-Amino-1-(3-(2-ethyl-2-
hydroxybutylamino)phenyl) propan-1-one 80 ##STR00151##
3-Amino-1-(3-((1- hydroxycyclopentyl) methylamino)
phenyl)propan-1-one 81 ##STR00152## 3-Amino-1-(3-
(cyclohexylmethylamino) phenyl)-1-deuteropropan-1-ol 82
##STR00153## 3-Amino-1-(3- (cyclohexylmethylamino) phenyl)-2,2-
dideuteropropan-1-ol 83 ##STR00154## 3-Amino-1-(3-
(cyclohexylmethylamino) phenyl)-3,3- dideuteropropan-1-ol 84
##STR00155## N-(3-(3-Amino-3,3-dideutero-1- hydroxypropyl)phenyl)
cyclohexanecarboxamide 85 ##STR00156##
N-(3-(3-Amino-3,3-dideutero-1- hydroxypropyl)phenyl)
cyclohexanesulfonamide 86 ##STR00157## (R)-3-Amino-1-(3-
(cyclohexylmethylamino) phenyl)propan-1-ol 87 ##STR00158##
3-Amino-1-(3- (cyclohexylmethylamino)phenyl)-2- methylpropan-1-ol
88 ##STR00159## 1-Amino-3-(3- (cyclohexylmethylamino)
phenyl)propan-2-ol 89 ##STR00160## N-(3-(3- (cyclohexylmethylamino)
phenyl)-3-hydroxypropyl) acetamide 90 ##STR00161## 3-Amino-1-(3-
((cyclohexylmethyl)(methyl) amino)phenyl)propan-1-ol 91
##STR00162## 3-Amino-1-(3-((1- deuterocyclohexyl) methylamino)
phenyl)propan-1-ol 92 ##STR00163## 3-Amino-1-(3-
cyclohexyldideutero- methylamino)phenyl) propan-1-ol 93
##STR00164## N-(3-(3-Amino-1- hydroxypropyl)phenyl)-
1,2,2,3,3,4,4,5,5,6,6- undecadeuterocyclo- hexanecarboxamide 94
##STR00165## 1-(3-(cyclohexylmethyl- amino)phenyl)-
3-(methylamino)propan-1-ol 95 ##STR00166## 3-(3-Aminopropyl)-N-
pentylaniline
96 ##STR00167## N-(3-(3- Aminopropyl)phenyl) pentanamide 97
##STR00168## N-(3-(3-Amino-1- hydroxypropyl)phenyl)
cyclopentanesulfonamide 98 ##STR00169## N-(3-(3-
Aminopropanoyl)phenyl) cyclopentanesulfonamide 99 ##STR00170##
N-(3-(3- Aminopropyl)phenyl) benzenesulfonamide 100 ##STR00171##
3-Amino-1-(3- (benzylamino)phenyl) propan-1-ol 101 ##STR00172##
N-(3-(3-Amino-1- hydroxypropyl)phenyl) benzenesulfonamide 102
##STR00173## 3-Amino-1-(3- (benzylamino)phenyl) propan-1-one 103
##STR00174## N-(3-(3- Aminopropanoyl)phenyl) benzenesulfonamide 104
##STR00175## 3-(3-Aminopropyl)-N-(2- methoxybenzyl)aniline 105
##STR00176## 3-(3-Aminopropyl)-N- phenethylaniline 106 ##STR00177##
3-(3-Aminopropyl)-N-(thiazol-2- ylmethyl)aniline 107 ##STR00178##
N-(3-(3-Aminopropyl) phenyl)-2-cyclohexylethane- sulfonamide 108
##STR00179## N-(3-(3-Aminopropanoyl) phenyl)-2-cyclohexylethane-
sulfonamide 109 ##STR00180## N-(3-(3-Amino-1-
hydroxypropyl)phenyl)-2- cyclohexylethane- sulfonamide 110
##STR00181## 3-(3-Aminopropyl)-N-(5- (benzyloxy)pentyl)aniline 111
##STR00182## N-(3-(3-Aminopropyl) phenyl)-5-methoxypentane-
1-sulfonamide 112 ##STR00183## N-(3-(3-Amino-1-
hydroxypropyl)phenyl)- 5-methoxypentane-1- sulfonamide 113
##STR00184## N-(3-(3-Aminopropanoyl) phenyl)-5-methoxypentane-
1-sulfonamide 114 ##STR00185## (E)-1-(3-(3-Amino-
1-fluoro-1-hydroxypropyl) styryl)cyclohexanol 115 ##STR00186##
(E)-3-amino-1-(3-(2- cyclohexylvinyl)phenyl)-2,2-
dideuteropropan-1-ol 116 ##STR00187## (E)-1-(3-(3-Amino-3,3-
dideutero-1-hydroxypropyl) styryl)cyclohexanol 117 ##STR00188##
(E)-4-(2-(3-(3-Amino-1- hydroxypropyl)phenyl)-1,2-
dideuterovinyl)heptan-4-ol 118 ##STR00189## (E)-1-(3-(3-Amino-1-
hydroxypropyl)-4- deuterostyryl)cyclohexanol 119 ##STR00190##
4-((3-(3-Amino-1-deutero-1- hydroxypropyl)phenyl)ethynyl)
heptan-4-ol 120 ##STR00191## 1-((3-(3-Amino-2,2-dideutero-1-
hydroxypropyl)phenypethynyl) cyclohexanol 121 ##STR00192##
1-((3-(3-Amino-3,3-dideutero-1- hydroxypropyl)phenypethynyl)
cyclohexanol 122 ##STR00193## 3-Amino-1-(3-
(cyclohexylethynyl)phenyl)-2,2- dideuteropropan-1-ol 123
##STR00194## 3-Amino-1-(3- (cyclohexylethynyl)phenyl)-3,3-
dideuteropropan-1-ol 124 ##STR00195## 1-((3-(3-Amino-1-
hydroxypropyl)-4- deuterophenyl)ethynyl) cyclohexanol 125
##STR00196## 1-((3-(3-Amino-1- hydroxypropyl)-5-
deuterophenyl)ethynyl) cyclohexanol 126 ##STR00197## 3-Amino-1-(3-
(cyclohexylmethoxy)phenyl)-1- deuteropropan-1-ol 127 ##STR00198##
3-Amino-1-(3- (cyclohexylmethoxy)phenyl)-2,2- dideuteropropan-1-ol
128 ##STR00199## 3-Amino-1-(3- (cyclohexylmethoxy)phenyl)-3,3-
dideuteropropan-1-ol 129 ##STR00200## 3-Amino-1-(3-((1-
deuterocyclohexyl)methoxy) phenyl)propan-1-ol 130 ##STR00201##
(R)-3-Amino-1-(3- (cyclohexyldideuteromethoxy) phenyl)propan-1-ol
131 ##STR00202## 3-Amino-1-(3- ((perdeuterocyclohexyl)methoxy)
phenyl)propan-1-ol 132 ##STR00203## 3-Amino-1-(3-
(cyclohexylmethoxy)-5- deuterophenyl)propan-1-ol
[0766] As used herein and in the appended claims, the singular
forms "a," "and," and "the" include plural referents unless the
context clearly dictates otherwise. Thus, for example, reference to
"a compound" includes a plurality of such compounds, and reference
to "the cell" includes reference to one or more cells (or to a
plurality of cells) and equivalents thereof known to those skilled
in the art, and so forth. When ranges are used herein for physical
properties, such as molecular weight, or chemical properties, such
as chemical formulae, all combinations and subcombinations of
ranges and specific embodiments therein are intended to be
included. The term "about" when referring to a number or a
numerical range means that the number or numerical range referred
to is an approximation within experimental variability (or within
statistical experimental error), and thus the number or numerical
range may vary between 1% and 15% of the stated number or numerical
range. The term "comprising" (and related terms such as "comprise"
or "comprises" or "having" or "including") is not intended to
exclude that in other certain embodiments, for example, an
embodiment of any composition of matter, composition, method, or
process, or the like, described herein, may "consist of" or
"consist essentially of" the described features.
"Sulfanyl" refers to the --S-- radical. "Sulfonyl" refers to the
--S(.dbd.O)-- radical. "Sulfonyl" refers to the --S(.dbd.O).sub.2--
radical. "Amino" refers to the --NH.sub.2 radical. "Cyano" refers
to the --CN radical. "Nitro" refers to the --NO.sub.2 radical.
"Oxa" refers to the --O-- radical. "Oxo" refers to the .dbd.O
radical. "Imino" refers to the .dbd.NH radical. "Thioxo" refers to
the .dbd.S radical.
[0767] "Alkyl" refers to a straight or branched hydrocarbon chain
radical consisting solely of carbon and hydrogen atoms, containing
no unsaturation, having from one to fifteen carbon atoms (e.g.,
C.sub.1-C.sub.15 alkyl). In certain embodiments, an alkyl comprises
one to thirteen carbon atoms (e.g., C.sub.1-C.sub.13 alkyl). In
certain embodiments, an alkyl comprises one to eight carbon atoms
(e.g., C.sub.1-C.sub.8 alkyl). In other embodiments, an alkyl
comprises five to fifteen carbon atoms (e.g., C.sub.5-C.sub.15
alkyl). In other embodiments, an alkyl comprises five to eight
carbon atoms (e.g., C.sub.5-C.sub.8 alkyl). The alkyl is attached
to the rest of the molecule by a single bond, for example, methyl
(Me), ethyl (Et), n-propyl, 1-methylethyl (iso-propyl), n-butyl,
n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl,
2-methylhexyl, and the like. Unless stated otherwise specifically
in the specification, an alkyl group is optionally substituted by
one or more of the following substituents: halo, cyano, nitro, oxo,
thioxo, trimethylsilanyl, --OR.sup.a, --SR.sup.a, --OC(O)--R.sup.a,
--N(R.sup.a).sub.2, --C(O)R.sup.a, --C(O)OR.sup.a,
--C(O)N(R.sup.a).sub.2, --N(R.sup.a)C(O)OR.sup.a,
--N(R.sup.a)C(O)R.sup.a, --N(R.sup.a)S(O).sub.tR.sup.a (where t is
1 or 2), --S(O).sub.tOR.sup.a (where t is 1 or 2) and
--S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2) where each R.sup.a
is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,
carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,
heteroaryl or heteroarylalkyl.
[0768] "Alkenyl" refers to a straight or branched hydrocarbon chain
radical group consisting solely of carbon and hydrogen atoms,
containing at least one double bond, and having from two to twelve
carbon atoms. In certain embodiments, an alkenyl comprises two to
eight carbon atoms. In other embodiments, an alkenyl comprises two
to four carbon atoms. The alkenyl is attached to the rest of the
molecule by a single bond, for example, ethenyl (i.e., vinyl),
prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl,
penta-1,4-dienyl, and the like. Unless stated otherwise
specifically in the specification, an alkenyl group is optionally
substituted by one or more of the following substituents: halo,
cyano, nitro, oxo, thioxo, trimethylsilanyl, --OR.sup.a,
--SR.sup.a, --OC(O)--R.sup.a, --N(R.sup.a).sub.2, --C(O)R.sup.a,
--C(O)OR.sup.a, --C(O)N(R.sup.a).sub.2, --N(R.sup.a)C(O)OR.sup.a,
--N(R.sup.a)C(O)R.sup.a, --N(R.sup.a)S(O).sub.tR.sup.a (where t is
1 or 2), --S(O).sub.tOR.sup.a (where t is 1 or 2) and
--S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2) where each R.sup.a
is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,
carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,
heteroaryl or heteroarylalkyl.
[0769] "Alkynyl" refers to a straight or branched hydrocarbon chain
radical group consisting solely of carbon and hydrogen atoms,
containing at least one triple bond, having from two to twelve
carbon atoms. In certain embodiments, an alkynyl comprises two to
eight carbon atoms. In other embodiments, an alkynyl has two to
four carbon atoms. The alkynyl is attached to the rest of the
molecule by a single bond, for example, ethynyl, propynyl, butynyl,
pentynyl, hexynyl, and the like. Unless stated otherwise
specifically in the specification, an alkynyl group is optionally
substituted by one or more of the following substituents: halo,
cyano, nitro, oxo, thioxo, trimethylsilanyl, --OR.sup.a,
--SR.sup.a, --OC(O)--R.sup.a, --N(R.sup.a).sub.2, --C(O)R.sup.a,
--C(O)ORa, --C(O)N(R.sup.a).sub.2, --N(R.sup.a)C(O)OR.sup.a,
--N(R.sup.a)C(O)R.sup.a, --N(R.sup.a)S(O).sub.tR.sup.a (where t is
1 or 2), --S(O).sub.tOR.sup.a (where t is 1 or 2) and
--S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2) where each R.sup.a
is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,
carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,
heteroaryl or heteroarylalkyl.
[0770] "Alkylene" or "alkylene chain" refers to a straight or
branched divalent hydrocarbon chain linking the rest of the
molecule to a radical group, consisting solely of carbon and
hydrogen, containing no unsaturation and having from one to twelve
carbon atoms, for example, methylene, ethylene, propylene,
n-butylene, and the like. The alkylene chain is attached to the
rest of the molecule through a single bond and to the radical group
through a single bond. The points of attachment of the alkylene
chain to the rest of the molecule and to the radical group can be
through one carbon in the alkylene chain or through any two carbons
within the chain. Unless stated otherwise specifically in the
specification, an alkylene chain is optionally substituted by one
or more of the following substituents: halo, cyano, nitro, aryl,
cycloalkyl, heterocyclyl, heteroaryl, oxo, thioxo,
trimethylsilanyl, --OR.sup.a, --SR.sup.a, --OC(O)--R.sup.a,
--N(R.sup.a).sub.2, --C(O)R.sup.a, --C(O)OR.sup.a,
--C(O)N(R.sup.a).sub.2, --N(R.sup.a)C(O)OR.sup.a,
--N(R.sup.a)C(O)R.sup.a, --N(R.sup.a)S(O).sub.tR.sup.a (where t is
1 or 2), --S(O).sub.tOR.sup.a (where t is 1 or 2) and
--S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2) where each R.sup.a
is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,
carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,
heteroaryl or heteroarylalkyl.
[0771] "Alkenylene" or "alkenylene chain" refers to a straight or
branched divalent hydrocarbon chain linking the rest of the
molecule to a radical group, consisting solely of carbon and
hydrogen, containing at least one double bond and having from two
to twelve carbon atoms, for example, ethenylene, propenylene,
n-butenylene, and the like. The alkenylene chain is attached to the
rest of the molecule through a double bond or a single bond and to
the radical group through a double bond or a single bond. The
points of attachment of the alkenylene chain to the rest of the
molecule and to the radical group can be through one carbon or any
two carbons within the chain. Unless stated otherwise specifically
in the specification, an alkenylene chain is optionally substituted
by one or more of the following substituents: halo, cyano, nitro,
aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, thioxo,
trimethylsilanyl, --OR.sup.a, --SR.sup.a, --OC(O)--R.sup.a,
--N(R.sup.a).sub.2, --C(O)R.sup.a, --C(O)OR.sup.a,
--C(O)N(R.sup.a).sub.2, --N(R.sup.a)C(O)OR.sup.a,
--N(R.sup.a)C(O)R.sup.a, --N(R.sup.a)S(O).sub.tR.sup.a (where t is
1 or 2), --S(O).sub.tOR.sup.a (where t is 1 or 2) and
--S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2) where each R.sup.a
is independently hydrogen, alkyl, fluoroalkyl, cycloalkyl,
cycloalkylalkyl, aryl (optionally substituted with one or more halo
groups), aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or
heteroarylalkyl, and where each of the above substituents is
unsubstituted unless otherwise indicated.
[0772] "Aryl" refers to a radical derived from an aromatic
monocyclic or multicyclic hydrocarbon ring system by removing a
hydrogen atom from a ring carbon atom. The aromatic monocyclic or
multicyclic hydrocarbon ring system contains only hydrogen and
carbon from six to eighteen carbon atoms, where at least one of the
rings in the ring system is fully unsaturated, i.e., it contains a
cyclic, delocalized (4n+2) .pi.-electron system in accordance with
the Huckel theory. Aryl groups include, but are not limited to,
groups such as phenyl, fluorenyl, and naphthyl. Unless stated
otherwise specifically in the specification, the term "aryl" or the
prefix "ar-" (such as in "aralkyl") is meant to include aryl
radicals optionally substituted by one or more substituents
independently selected from alkyl, alkenyl, alkynyl, halo,
fluoroalkyl, cyano, nitro, optionally substituted aryl, optionally
substituted aralkyl, optionally substituted aralkenyl, optionally
substituted aralkynyl, optionally substituted carbocyclyl,
optionally substituted carbocyclylalkyl, optionally substituted
heterocyclyl, optionally substituted heterocyclylalkyl, optionally
substituted heteroaryl, optionally substituted heteroarylalkyl,
--R.sup.b--OR.sup.a,--R.sup.b--OC(O)--R.sup.a,
--R.sup.b--N(R.sup.a).sub.2, --R.sup.b--C(O)R.sup.a,
R.sup.b--C(O)OR.sup.a, --R.sup.b--C(O)N(R.sup.a).sub.2,
--R.sup.b--O--R.sup.c--C(O)N(R.sup.a).sub.2,
--R.sup.b--N(R.sup.a)C(O)OR.sup.a,
--R.sup.b--N(R.sup.a)C(O)R.sup.a,
--R.sup.b--N(R.sup.a)S(O).sub.tR.sup.a (where t is 1 or 2),
--R.sup.b--S(O).sub.tOR.sup.a (where t is 1 or 2) and
--R.sup.b--S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2), where
each R.sup.a is independently hydrogen, alkyl, fluoroalkyl,
cycloalkyl, cycloalkylalkyl, aryl (optionally substituted with one
or more halo groups), aralkyl, heterocyclyl, heterocyclylalkyl,
heteroaryl or heteroarylalkyl, each R.sup.b is independently a
direct bond or a straight or branched alkylene or alkenylene chain,
and R.sup.c is a straight or branched alkylene or alkenylene chain,
and where each of the above substituents is unsubstituted unless
otherwise indicated.
[0773] "Aralkyl" refers to a radical of the formula --R.sup.c-aryl
where R.sup.c is an alkylene chain as defined above, for example,
benzyl, diphenylmethyl and the like. The alkylene chain part of the
aralkyl radical is optionally substituted as described above for an
alkylene chain. The aryl part of the aralkyl radical is optionally
substituted as described above for an aryl group.
[0774] "Aralkenyl" refers to a radical of the formula
--R.sup.d-aryl where R.sup.d is an alkenylene chain as defined
above. The aryl part of the aralkenyl radical is optionally
substituted as described above for an aryl group. The alkenylene
chain part of the aralkenyl radical is optionally substituted as
defined above for an alkenylene group.
[0775] "Aralkynyl" refers to a radical of the formula --Re-aryl,
where R.sup.e is an alkynylene chain as defined above. The aryl
part of the aralkynyl radical is optionally substituted as
described above for an aryl group. The alkynylene chain part of the
aralkynyl radical is optionally substituted as defined above for an
alkynylene chain.
[0776] "Carbocyclyl" refers to a stable non-aromatic monocyclic or
polycyclic hydrocarbon radical consisting solely of carbon and
hydrogen atoms, which includes fused or bridged ring systems,
having from three to fifteen carbon atoms. In certain embodiments,
a carbocyclyl comprises three to ten carbon atoms. In other
embodiments, a carbocyclyl comprises five to seven carbon atoms.
The carbocyclyl is attached to the rest of the molecule by a single
bond. Carbocyclyl is optionally saturated, (i.e., containing single
C--C bonds only) or unsaturated (i.e., containing one or more
double bonds or triple bonds.) A fully saturated carbocyclyl
radical is also referred to as "cycloalkyl." Examples of monocyclic
cycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl, and cyclooctyl. An unsaturated carbocyclyl
is also referred to as "cycloalkenyl." Examples of monocyclic
cycloalkenyls include, e.g., cyclopentenyl, cyclohexenyl,
cycloheptenyl, and cyclooctenyl. Polycyclic carbocyclyl radicals
include, for example, adamantyl, norbornyl (i.e.,
bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl,
7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise
stated specifically in the specification, the term "carbocyclyl" is
meant to include carbocyclyl radicals that are optionally
substituted by one or more substituents independently selected from
alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano,
nitro, optionally substituted aryl, optionally substituted aralkyl,
optionally substituted aralkenyl, optionally substituted aralkynyl,
optionally substituted carbocyclyl, optionally substituted
carbocyclylalkyl, optionally substituted heterocyclyl, optionally
substituted heterocyclylalkyl, optionally substituted heteroaryl,
optionally substituted heteroarylalkyl, --R.sup.b--OR.sup.a,
--R.sup.b--SR.sup.a, --R.sup.b--OC(O)--R.sup.a,
--R.sup.b--N(R.sup.a).sub.2, --R.sup.b--C(O)R.sup.a,
--R.sup.b--C(O)OR.sup.a, --R.sup.b--C(O)N(R.sup.a).sub.2,
--R.sup.b--O--R.sup.c--C(O)N(R.sup.a).sub.2,
--R.sup.b--N(R.sup.a)C(O)OR.sup.a,
--R.sup.b--N(R.sup.a)C(O)R.sup.a,
--R.sup.b--N(R.sup.a)S(O).sub.tR.sup.a (where t is 1 or 2),
--R.sup.b--S(O).sub.tOR.sup.a (where t is 1 or 2) and
--R.sup.b--S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2), where
each R.sup.a is independently hydrogen, alkyl, fluoroalkyl,
cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl,
heterocyclylalkyl, heteroaryl or heteroarylalkyl, each R.sup.b is
independently a direct bond or a straight or branched alkylene or
alkenylene chain, and R.sup.c is a straight or branched alkylene or
alkenylene chain, and where each of the above substituents is
unsubstituted unless otherwise indicated.
[0777] "Carbocyclylalkyl" refers to a radical of the formula
--R.sup.c-carbocyclyl where R.sup.c is an alkylene chain as defined
above. The alkylene chain and the carbocyclyl radical is optionally
substituted as defined above.
[0778] "Halo" or "halogen" refers to bromo, chloro, fluoro or iodo
substituents.
[0779] "Fluoroalkyl" refers to an alkyl radical, as defined above,
that is substituted by one or more fluoro radicals, as defined
above, for example, trifluoromethyl, difluoromethyl,
2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like.
The alkyl part of the fluoroalkyl radical is optionally substituted
as defined above for an alkyl group.
[0780] "Heterocyclyl" refers to a stable 3- to 18-membered
non-aromatic ring radical that comprises two to twelve carbon atoms
and from one to six heteroatoms selected from nitrogen, oxygen and
sulfur. Unless stated otherwise specifically in the specification,
the heterocyclyl radical is a monocyclic, bicyclic, tricyclic or
tetracyclic ring system, and includes fused or bridged ring
systems. The heteroatom(s) in the heterocyclyl radical is
optionally oxidized. One or more nitrogen atoms, if present, are
optionally quaternized. The heterocyclyl radical is partially or
fully saturated. The heterocyclyl is attached to the rest of the
molecule through any atom of the ring(s). Examples of such
heterocyclyl radicals include, but are not limited to, dioxolanyl,
thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,
imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,
octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,
2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,
piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl,
quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl,
tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl,
1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated
otherwise specifically in the specification, the term
"heterocyclyl" is meant to include heterocyclyl radicals as defined
above that are optionally substituted by one or more substituents
selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo,
thioxo, cyano, nitro, optionally substituted aryl, optionally
substituted aralkyl, optionally substituted aralkenyl, optionally
substituted aralkynyl, optionally substituted carbocyclyl,
optionally substituted carbocyclylalkyl, optionally substituted
heterocyclyl, optionally substituted heterocyclylalkyl, optionally
substituted heteroaryl, optionally substituted heteroarylalkyl,
--R.sup.b--OR.sup.a, --R.sup.b--SR.sup.a,
--R.sup.b--OC(O)--R.sup.a, --R.sup.b--N(R.sup.a).sub.2,
--R.sup.b--C(O)R.sup.a, --R.sup.b--C(O)OR.sup.a,
--R.sup.b--C(O)N(R.sup.a).sub.2,
--R.sup.b--O--R.sup.c--C(O)N(R.sup.a).sub.2,
--R.sup.b--N(R.sup.a)C(O)OR.sup.a,
--R.sup.b--N(R.sup.a)C(O)R.sup.a,
--R.sup.b--N(R.sup.a)S(O).sub.tR.sup.a (where t is 1 or 2),
--R.sup.b--S(O).sub.tOR.sup.a (where t is 1 or 2) and
--R.sup.b--S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2), where
each R.sup.a is independently hydrogen, alkyl, fluoroalkyl,
cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl,
heterocyclylalkyl, heteroaryl or heteroarylalkyl, each R.sup.b is
independently a direct bond or a straight or branched alkylene or
alkenylene chain, and R.sup.c is a straight or branched alkylene or
alkenylene chain, and where each of the above substituents is
unsubstituted unless otherwise indicated.
[0781] "N-heterocyclyl" or "N-attached heterocyclyl" refers to a
heterocyclyl radical as defined above containing at least one
nitrogen and where the point of attachment of the heterocyclyl
radical to the rest of the molecule is through a nitrogen atom in
the heterocyclyl radical. An N-heterocyclyl radical is optionally
substituted as described above for heterocyclyl radicals. Examples
of such N-heterocyclyl radicals include, but are not limited to,
1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl,
pyrazolidinyl, imidazolinyl, and imidazolidinyl.
[0782] "C-heterocyclyl" or "C-attached heterocyclyl" refers to a
heterocyclyl radical as defined above containing at least one
heteroatom and where the point of attachment of the heterocyclyl
radical to the rest of the molecule is through a carbon atom in the
heterocyclyl radical. A C-heterocyclyl radical is optionally
substituted as described above for heterocyclyl radicals. Examples
of such C-heterocyclyl radicals include, but are not limited to,
2-morpholinyl, 2- or 3- or 4-piperidinyl, 2-piperazinyl, 2- or
3-pyrrolidinyl, and the like.
[0783] "Heterocyclylalkyl" refers to a radical of the formula
--R.sup.c-heterocyclyl where R.sup.c is an alkylene chain as
defined above. If the heterocyclyl is a nitrogen-containing
heterocyclyl, the heterocyclyl is optionally attached to the alkyl
radical at the nitrogen atom. The alkylene chain of the
heterocyclylalkyl radical is optionally substituted as defined
above for an alkylene chain. The heterocyclyl part of the
heterocyclylalkyl radical is optionally substituted as defined
above for a heterocyclyl group.
[0784] "Heteroaryl" refers to a radical derived from a 3- to
18-membered aromatic ring radical that comprises two to seventeen
carbon atoms and from one to six heteroatoms selected from
nitrogen, oxygen and sulfur. As used herein, the heteroaryl radical
is a monocyclic, bicyclic, tricyclic or tetracyclic ring system,
wherein at least one of the rings in the ring system is fully
unsaturated, i.e., it contains a cyclic, delocalized (4n+2)
.pi.-electron system in accordance with the Huckel theory.
Heteroaryl includes fused or bridged ring systems. The
heteroatom(s) in the heteroaryl radical is optionally oxidized. One
or more nitrogen atoms, if present, are optionally quaternized. The
heteroaryl is attached to the rest of the molecule through any atom
of the ring(s). Examples of heteroaryls include, but are not
limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl,
1,3-benzodioxolyl, benzofuranyl, benzooxazolyl, benzo[d]thiazolyl,
benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl,
1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl,
benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl,
benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl),
benzothieno[3,2-d]pyrimidinyl, benzotriazolyl,
benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,
cyclopenta[d]pyrimidinyl,
6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,
5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl,
6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl,
dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl,
furo[3,2-c]pyridinyl,
5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,
5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl,
5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl,
imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl,
isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl,
5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl,
1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl,
oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl,
1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl,
phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl,
pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl,
pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl,
pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl,
tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl,
5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl,
6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,
5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl,
thiadiazolyl, triazolyl, tetrazolyl, triazinyl,
thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl,
thieno[2,3-c]pridinyl, and thiophenyl (i.e. thienyl). Unless stated
otherwise specifically in the specification, the term "heteroaryl"
is meant to include heteroaryl radicals as defined above which are
optionally substituted by one or more substituents selected from
alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl,
haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted
aryl, optionally substituted aralkyl, optionally substituted
aralkenyl, optionally substituted aralkynyl, optionally substituted
carbocyclyl, optionally substituted carbocyclylalkyl, optionally
substituted heterocyclyl, optionally substituted heterocyclylalkyl,
optionally substituted heteroaryl, optionally substituted
heteroarylalkyl, --R.sup.b--OR.sup.a, --R.sup.b--SR.sup.a,
--R.sup.b--OC(O)--R.sup.a, --R.sup.b--N(R.sup.a).sub.2,
--R.sup.b--C(O)R.sup.a, --R.sup.b--C(O)OR.sup.a,
--R.sup.b--C(O)N(R.sup.a).sub.2,
--R.sup.b--O--R.sup.c--C(O)N(R.sup.a).sub.2,
--R.sup.b--N(R.sup.a)C(O)OR.sup.a,
--R.sup.b--N(R.sup.a)C(O)R.sup.a,
--R.sup.b--N(R.sup.a)S(O).sub.tR.sup.a (where t is 1 or 2),
--R.sup.b--S(O).sub.tOR.sup.a (where t is 1 or 2) and
--R.sup.b--S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2), where
each R.sup.a is independently hydrogen, alkyl, fluoroalkyl,
cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl,
heterocyclylalkyl, heteroaryl or heteroarylalkyl, each R.sup.b is
independently a direct bond or a straight or branched alkylene or
alkenylene chain, and R.sup.c is a straight or branched alkylene or
alkenylene chain, and where each of the above substituents is
unsubstituted unless otherwise indicated.
[0785] "N-heteroaryl" refers to a heteroaryl radical as defined
above containing at least one nitrogen and where the point of
attachment of the heteroaryl radical to the rest of the molecule is
through a nitrogen atom in the heteroaryl radical. An N-heteroaryl
radical is optionally substituted as described above for heteroaryl
radicals.
[0786] "C-heteroaryl" refers to a heteroaryl radical as defined
above and where the point of attachment of the heteroaryl radical
to the rest of the molecule is through a carbon atom in the
heteroaryl radical. A C-heteroaryl radical is optionally
substituted as described above for heteroaryl radicals.
[0787] "Heteroarylalkyl" refers to a radical of the formula
--R.sup.c-heteroaryl, where R.sup.c is an alkylene chain as defined
above. If the heteroaryl is a nitrogen-containing heteroaryl, the
heteroaryl is optionally attached to the alkyl radical at the
nitrogen atom. The alkylene chain of the heteroarylalkyl radical is
optionally substituted as defined above for an alkylene chain. The
heteroaryl part of the heteroarylalkyl radical is optionally
substituted as defined above for a heteroaryl group.
[0788] The compounds, or their pharmaceutically acceptable salts
may contain one or more asymmetric centers and may thus give rise
to enantiomers, diastereomers, and other stereoisomeric forms that
may be defined, in terms of absolute stereochemistry, as (R)- or
(S)- or, as (D)- or (L)- for amino acids. When the compounds
described herein contain olefinic double bonds or other centers of
geometric asymmetry, and unless specified otherwise, it is intended
that the compounds include both E and Z geometric isomers (e.g.,
cis or trans.) Likewise, all possible isomers, as well as their
racemic and optically pure forms, and all tautomeric forms are also
intended to be included.
[0789] "Stereoisomers" are compounds that have the same sequence of
covalent bonds and differ in the relative disposition of their
atoms in space. "Enantiomers" refers to two stereoisomers that are
nonsuperimposeable mirror images of one another.
[0790] Unless otherwise stated, structures depicted herein are also
meant to include compounds which differ only in the presence of one
or more isotopically enriched atoms. For example, compounds having
the present structures except for the replacement of a hydrogen by
a deuterium or tritium, or the replacement of a carbon by .sup.13C-
or .sup.14C-enriched carbon are within the scope of this
invention.
[0791] The compounds of the present invention may also contain
unnatural proportions of atomic isotopes at one or more atoms that
constitute such compounds. For example, the compounds may be
labeled with isotopes, such as for example, deuterium (.sup.2H),
tritium (.sup.3H), iodine-125 (.sup.125I) or carbon-14 (.sup.14C).
Isotopic substitution with .sup.2H, .sup.11C, .sup.13C, .sup.14C,
.sup.15C, .sup.12N, .sup.13N, .sup.15N, .sup.16N, .sup.16O,
.sup.17O, .sup.14F, .sup.15F, .sup.16F, .sup.17F, .sup.18F,
.sup.33S, .sup.34S, .sup.35S, .sup.36S, .sup.35Cl, .sup.37Cl,
.sup.79Br, .sup.81Br, .sup.125I are all contemplated. All isotopic
variations of the compounds of the present invention, whether
radioactive or not, are encompassed within the scope of the present
invention.
[0792] In certain embodiments, the compounds disclosed herein have
some or all of the .sup.1H atoms replaced with .sup.2H atoms. The
methods of synthesis for deuterium-containing amine derivative
compounds are known in the art and include, by way of non-limiting
example only, the following synthetic methods.
[0793] Deuterated starting materials, such as acid i and acid ii,
are readily available and are subjected to the synthetic methods
described herein for the synthesis of amine derivative
compounds.
##STR00204##
[0794] Other deuterated starting materials are also employed in the
synthesis of deuterium-containing amine derivative compounds as
shown, in a non-limiting example, in the scheme below. Large
numbers of deuterium-containing reagents and building blocks are
available commerically from chemical vendors, such as Aldrich
Chemical Co.
##STR00205##
[0795] Deuterium-transfer reagents, such as lithium aluminum
deuteride (LiAlD.sub.4), are employed to transfer deuterium under
reducing conditions to the reaction substrate. The use of
LiAlD.sub.4 is illustrated, by way of example only, in the reaction
schemes below.
##STR00206##
[0796] Deuterium gas and palladium catalyst are employed to reduce
unsaturated carbon-carbon linkages and to perform a reductive
substitution of aryl carbon-halogen bonds as illustrated, by way of
example only, in the reaction schemes below.
##STR00207##
[0797] In one embodiments, the compounds disclosed herein contain
one deuterium atom. In another embodiment, the compounds disclosed
herein contains two deuterium atoms. In another embodiment, the
compounds disclosed herein contains three deuterium atoms. In
another embodiment, the compounds disclosed herein contains four
deuterium atoms. In another embodiment, the compounds disclosed
herein contains five deuterium atoms. In another embodiment, the
compounds disclosed herein contains six deuterium atoms. In another
embodiment, the compounds disclosed herein contains more than six
deuterium atoms. In another embodiment, the compounds disclosed
herein is fully substituted with deuterium atoms and contains no
non-exchangeable .sup.1H hydrogen atoms. In one embodiment, the
level of deuterium incorporation is determined by synthetic methods
in which a per-deuterated synthetic building block is used as a
starting material. In one embodiment, acid ii is incorporated in
the compounds disclosed herein to provide a compound with eleven
deuterium atoms such as, by way of example only, compound iii.
##STR00208##
[0798] In another embodiment, is a deuterium labeled compound
selected from:
##STR00209## ##STR00210## ##STR00211## ##STR00212##
[0799] Compounds described herein optionally have a substitution of
one, more than one or all of the non-exchangeable hydrogen atoms
for deuterium atoms. A non-exchangeable hydrogen atom is one bound
to a carbon atom. This type of deuterium substitution provides for
improved pharmacokinetic and pharmacodynamic properties. As the C-D
bond is stronger than the C--H bond, a metabolic process that
involves breaking a C--H bond will be relatively slower for the C-D
analog. Pharmacokinetic and pharmacodynamic properties modulated by
deuterium substitution include bioavailability, serum half-life,
clearance, drug-drug interactions, CYP inhibition and metabolite
profile.
[0800] A "tautomer" refers to a proton shift from one atom of a
molecule to another atom of the same molecule. The compounds
presented herein may exist as tautomers. Tautomers are compounds
that are interconvertible by migration of a hydrogen atom,
accompanied by a switch of a single bond and adjacent double bond.
In bonding arrangements where tautomerization is possible, a
chemical equilibrium of the tautomers will exist. All tautomeric
forms of the compounds disclosed herein are contemplated. The exact
ratio of the tautomers depends on several factors, including
temperature, solvent, and pH. Some examples of tautomeric
interconversions include:
##STR00213##
[0801] "Optional" or "optionally" means that a subsequently
described event or circumstance may or may not occur and that the
description includes instances when the event or circumstance
occurs and instances in which it does not. For example, "optionally
substituted aryl" means that the aryl radical may or may not be
substituted and that the description includes both substituted aryl
radicals and aryl radicals having no substitution.
[0802] "Pharmaceutically acceptable salt" includes both acid and
base addition salts. A pharmaceutically acceptable salt of any one
of the compounds described herein is intended to encompass any and
all pharmaceutically suitable salt forms. Preferred
pharmaceutically acceptable salts of the compounds described herein
are pharmaceutically acceptable acid addition salts and
pharmaceutically acceptable base addition salts.
[0803] "Pharmaceutically acceptable acid addition salt" refers to
those salts which retain the biological effectiveness and
properties of the free bases, which are not biologically or
otherwise undesirable, and which are formed with inorganic acids
such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric
acid, phosphoric acid, hydroiodic acid, hydrofluoric acid,
phosphorous acid, and the like. Also included are salts that are
formed with organic acids such as aliphatic mono- and dicarboxylic
acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids,
alkanedioic acids, aromatic acids, aliphatic and. aromatic sulfonic
acids, etc. and include, for example, acetic acid, trifluoroacetic
acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid,
maleic acid, malonic acid, succinic acid, fumaric acid, tartaric
acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,
methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,
salicylic acid, and the like. Exemplary salts thus include
sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates,
phosphates, monohydrogenphosphates, dihydrogenphosphates,
metaphosphates, pyrophosphates, chlorides, bromides, iodides,
acetates, trifluoroacetates, propionates, caprylates, isobutyrates,
oxalates, malonates, succinate suberates, sebacates, fumarates,
maleates, mandelates, benzoates, chlorobenzoates, methylbenzoates,
dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates,
phenylacetates, citrates, lactates, malates, tartrates,
methanesulfonates, and the like. Also contemplated are salts of
amino acids, such as arginates, gluconates, and galacturonates
(see, for example, Berge S. M. et al., "Pharmaceutical Salts,"
Journal of Pharmaceutical Science, 66:1-19 (1997), which is hereby
incorporated by reference in its entirety). Acid addition salts of
basic compounds may be prepared by contacting the free base forms
with a sufficient amount of the desired acid to produce the salt
according to methods and techniques with which a skilled artisan is
familiar.
[0804] "Pharmaceutically acceptable base addition salt" refers to
those salts that retain the biological effectiveness and properties
of the free acids, which are not biologically or otherwise
undesirable. These salts are prepared from addition of an inorganic
base or an organic base to the free acid. Pharmaceutically
acceptable base addition salts may be formed with metals or amines,
such as alkali and alkaline earth metals or organic amines. Salts
derived from inorganic bases include, but are not limited to,
sodium, potassium, lithium, ammonium, calcium, magnesium, iron,
zinc, copper, manganese, aluminum salts and the like. Salts derived
from organic bases include, but are not limited to, salts of
primary, secondary, and tertiary amines, substituted amines
including naturally occurring substituted amines, cyclic amines and
basic ion exchange resins, for example, isopropylamine,
trimethylamine, diethylamine, triethylamine, tripropylamine,
ethanolamine, diethanolamine, 2-dimethylaminoethanol,
2-diethylaminoethanol, dicyclohexylamine, lysine, arginine,
histidine, caffeine, procaine, N,N-dibenzylethylenediamine,
chloroprocaine, hydrabamine, choline, betaine, ethylenediamine,
ethylenedianiline, N-methylglucamine, glucosamine, methylglucamine,
theobromine, purines, piperazine, piperidine, N-ethylpiperidine,
polyamine resins and the like. See Berge et al., supra.
[0805] Unless otherwise stated, structures depicted herein are also
meant to include compounds which differ only in the presence of one
or more isotopically enriched atoms. For example, compounds having
the present structures except for the replacement of a hydrogen by
a deuterium or tritium, or the replacement of a carbon by .sup.13C-
or .sup.14C-enriched carbon are within the scope of this
invention.
[0806] The compounds of the present invention may also contain
unnatural proportions of atomic isotopes at one or more of atoms
that constitute such compounds. For example, the compounds may be
radiolabeled with radioactive isotopes, such as for example tritium
(.sup.3H), iodine-125 (.sup.125I) or carbon-14 (.sup.14C). All
isotopic variations of the compounds of the present invention,
whether radioactive or not, are encompassed within the scope of the
present invention.
[0807] "Non-retinoid compound" refers to any compound that is not a
retinoid. A retinoid is a compound that has a diterpene skeleton
possessing a trimethylcyclohexenyl ring and a polyene chain that
terminates in a polar end group. Examples of retinoids include
retinaldehyde and derived imine/hydrazide/oxime, retinol and any
derived ester, retinol amine and any derived amide, retinoic acid
and any derived ester or amide. A non-retinoid compound can
comprise though not require an internal cyclic group (e.g.,
aromatic group). A non-retinoid compound can contain though not
require a nitrogen-linked group.
[0808] As used herein, "treatment" or "treating," or "palliating"
or "ameliorating" are used interchangeably herein. These terms
refers to an approach for obtaining beneficial or desired results
including but not limited to therapeutic benefit and/or a
prophylactic benefit. By therapeutic benefit is meant eradication
or amelioration of the underlying disorder being treated. Also, a
therapeutic benefit is achieved with the eradication or
amelioration of one or more of the physiological symptoms
associated with the underlying disorder such that an improvement is
observed in the patient, notwithstanding that the patient may still
be afflicted with the underlying disorder. For prophylactic
benefit, the compositions may be administered to a patient at risk
of developing a particular disease, or to a patient reporting one
or more of the physiological symptoms of a disease, even though a
diagnosis of this disease may not have been made.
[0809] "Prodrug" is meant to indicate a compound that may be
converted under physiological conditions or by solvolysis to a
biologically active compound described herein. Thus, the term
"prodrug" refers to a precursor of a biologically active compound
that is pharmaceutically acceptable. A prodrug may be inactive when
administered to a subject, but is converted in vivo to an active
compound, for example, by hydrolysis. The prodrug compound often
offers advantages of solubility, tissue compatibility or delayed
release in a mammalian organism (see, e.g., Bundgard, H., Design of
Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam).
[0810] A discussion of prodrugs is provided in Higuchi, T., et al.,
"Pro-drugs as Novel Delivery Systems," A.C.S. Symposium Series,
Vol. 14, and in Bioreversible Carriers in Drug Design, ed. Edward
B. Roche, American Pharmaceutical Association and Pergamon Press,
1987, both of which are incorporated in full by reference
herein.
[0811] The term "prodrug" is also meant to include any covalently
bonded carriers, which release the active compound in vivo when
such prodrug is administered to a mammalian subject. Prodrugs of an
active compound, as described herein, may be prepared by modifying
functional groups present in the active compound in such a way that
the modifications are cleaved, either in routine manipulation or in
vivo, to the parent active compound. Prodrugs include compounds
wherein a hydroxy, amino or mercapto group is bonded to any group
that, when the prodrug of the active compound is administered to a
mammalian subject, cleaves to form a free hydroxy, free amino or
free mercapto group, respectively. Examples of prodrugs include,
but are not limited to, acetate, formate and benzoate derivatives
of an alcohol or acetamide, formamide and benzamide derivatives of
an amine functional group in the active compound and the like.
[0812] The compounds of the invention are synthesized by an
appropriate combination of generally well known synthetic methods.
Techniques useful in synthesizing the compounds of the invention
are both readily apparent and accessible to those of skill in the
relevant art.
[0813] The discussion below is offered to illustrate how, in
principle, to gain access to the compounds claimed under this
invention and to give details on certain of the diverse methods
available for use in assembling the compounds of the invention.
However, the discussion is not intended to define or limit the
scope of reactions or reaction sequences that are useful in
preparing the compounds of the present invention. The compounds of
this invention may be made by the procedures and techniques
disclosed in the Examples section below, as well as by known
organic synthesis techniques.
I. Preparation of Compounds
[0814] In general, the compounds used in the reactions described
herein may be made according to organic synthesis techniques known
to those skilled in this art, starting from commercially available
chemicals and/or from compounds described in the chemical
literature. "Commercially available chemicals" may be obtained from
standard commercial sources including Acros Organics (Pittsburgh
Pa.), Aldrich Chemical (Milwaukee Wis., including Sigma Chemical
and Fluka), Apin Chemicals Ltd. (Milton Park UK), Avocado Research
(Lancashire U.K.), BDH Inc. (Toronto, Canada), Bionet (Cornwall,
U.K.), Chemservice Inc. (West Chester Pa.), Crescent Chemical Co.
(Hauppauge N.Y.), Eastman Organic Chemicals, Eastman Kodak Company
(Rochester N.Y.), Fisher Scientific Co. (Pittsburgh Pa.), Fisons
Chemicals (Leicestershire UK), Frontier Scientific (Logan Utah),
ICN Biomedicals, Inc. (Costa Mesa Calif.), Key Organics (Cornwall
U.K.), Lancaster Synthesis (Windham N.H.), Maybridge Chemical Co.
Ltd. (Cornwall U.K.), Parish Chemical Co. (Orem Utah), Pfaltz &
Bauer, Inc. (Waterbury Conn.), Polyorganix (Houston Tex.), Pierce
Chemical Co. (Rockford Ill.), Riedel de Haen A G (Hanover,
Germany), Spectrum Quality Product, Inc. (New Brunswick, N.J.), TCI
America (Portland Oreg.), Trans World Chemicals, Inc. (Rockville
Md.), and Wako Chemicals USA, Inc. (Richmond Va.).
[0815] Methods known to one of ordinary skill in the art may be
identified through various reference books and databases. Suitable
reference books and treatise that detail the synthesis of reactants
useful in the preparation of compounds described herein, or provide
references to articles that describe the preparation, include for
example, "Synthetic Organic Chemistry", John Wiley & Sons,
Inc., New York; S. R. Sandler et al., "Organic Functional Group
Preparations," 2nd Ed., Academic Press, New York, 1983; H. O.
House, "Modern Synthetic Reactions", 2nd Ed., W. A. Benjamin, Inc.
Menlo Park, Calif. 1972; T. L. Gilchrist, "Heterocyclic Chemistry",
2nd Ed., John Wiley & Sons, New York, 1992; J. March, "Advanced
Organic Chemistry: Reactions, Mechanisms and Structure", 4th Ed.,
Wiley-Interscience, New York, 1992. Additional suitable reference
books and treatise that detail the synthesis of reactants useful in
the preparation of compounds described herein, or provide
references to articles that describe the preparation, include for
example, Fuhrhop, J. and Penzlin G. "Organic Synthesis: Concepts,
Methods, Starting Materials", Second, Revised and Enlarged Edition
(1994) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R. V.
"Organic Chemistry, An Intermediate Text" (1996) Oxford University
Press, ISBN 0-19-509618-5; Larock, R. C. "Comprehensive Organic
Transformations: A Guide to Functional Group Preparations" 2nd
Edition (1999) Wiley-VCH, ISBN: 0-471-19031-4; March, J. "Advanced
Organic Chemistry: Reactions, Mechanisms, and Structure" 4th
Edition (1992) John Wiley & Sons, ISBN: 0-471-60180-2; Otera,
J. (editor) "Modern Carbonyl Chemistry" (2000) Wiley-VCH, ISBN:
3-527-29871-1; Patai, S. "Patai's 1992 Guide to the Chemistry of
Functional Groups" (1992) Interscience ISBN: 0-471-93022-9; Quin,
L. D. et al. "A Guide to Organophosphorus Chemistry" (2000)
Wiley-Interscience, ISBN: 0-471-31824-8; Solomons, T. W. G.
"Organic Chemistry" 7th Edition (2000) John Wiley & Sons, ISBN:
0-471-19095-0; Stowell, J. C., "Intermediate Organic Chemistry" 2nd
Edition (1993) Wiley-Interscience, ISBN: 0-471-57456-2; "Industrial
Organic Chemicals: Starting Materials and Intermediates: An
Ullmann's Encyclopedia" (1999) John Wiley & Sons, ISBN:
3-527-29645-X, in 8 volumes; "Organic Reactions" (1942-2000) John
Wiley & Sons, in over 55 volumes; and "Chemistry of Functional
Groups" John Wiley & Sons, in 73 volumes.
[0816] Specific and analogous reactants may also be identified
through the indices of known chemicals prepared by the Chemical
Abstract Service of the American Chemical Society, which are
available in most public and university libraries, as well as
through on-line databases (the American Chemical Society,
Washington, D.C., may be contacted for more details). Chemicals
that are known but not commercially available in catalogs may be
prepared by custom chemical synthesis houses, where many of the
standard chemical supply houses (e.g., those listed above) provide
custom synthesis services. A reference for the preparation and
selection of pharmaceutical salts of the compounds described herein
is P. H. Stahl & C. G. Wermuth "Handbook of Pharmaceutical
Salts", Verlag Helvetica Chimica Acta, Zurich, 2002.
[0817] The term "protecting group" refers to chemical moieties that
block some or all reactive moieties of a compound and prevent such
moieties from participating in chemical reactions until the
protective group is removed, for example, those moieties listed and
described in T. W. Greene, P. G. M. Wuts, Protective Groups in
Organic Synthesis, 3rd ed. John Wiley & Sons (1999). It may be
advantageous, where different protecting groups are employed, that
each (different) protective group be removable by a different
means. Protective groups that are cleaved under totally disparate
reaction conditions allow differential removal of such protecting
groups. For example, protective groups can be removed by acid,
base, and hydrogenolysis. Groups such as trityl, dimethoxytrityl,
acetal and tert-butyldimethylsilyl are acid labile and may be used
to protect carboxy and hydroxy reactive moieties in the presence of
amino groups protected with Cbz groups, which are removable by
hydrogenolysis, and Fmoc groups, which are base labile. Carboxylic
acid moieties may be blocked with base labile groups such as,
without limitation, methyl, or ethyl, and hydroxy reactive moieties
may be blocked with base labile groups such as acetyl in the
presence of amines blocked with acid labile groups such as
tert-butyl carbamate or with carbamates that are both acid and base
stable but hydrolytically removable.
[0818] Carboxylic acid and hydroxy reactive moieties may also be
blocked with hydrolytically removable protective groups such as the
benzyl group, while amine groups may be blocked with base labile
groups such as Fmoc. Carboxylic acid reactive moieties may be
blocked with oxidatively-removable protective groups such as
2,4-dimethoxybenzyl, while co-existing amino groups may be blocked
with fluoride labile silyl carbamates.
[0819] Allyl blocking groups are useful in the presence of acid-
and base-protecting groups since the former are stable and can be
subsequently removed by metal or pi-acid catalysts. For example, an
allyl-blocked carboxylic acid can be deprotected with a
palladium(0)-catalyzed reaction in the presence of acid labile
t-butyl carbamate or base-labile acetate amine protecting groups.
Yet another form of protecting group is a resin to which a compound
or intermediate may be attached. As long as the residue is attached
to the resin, that functional group is blocked and cannot react.
Once released from the resin, the functional group is available to
react.
[0820] Typical blocking/protecting groups are known in the art and
include, but are not limited to the following moieties.
##STR00214##
Methods for Preparing Compounds of Formula (I)
[0821] The following methods illustrate various synthetic pathways
for preparing nitogen-linked intermediates and the side chain
moieties. One skilled in the art will recognize that, for example,
a method for amide formation can be combined with a method for side
chain formation. For example, any one of Methods A-C can be
combined with any of Methods D-H, or any of Methods I-J. They can
be further combined with any of Methods K-S to modify the linkage
and/or the terminal nitrogen-containing moiety. In the following
methods Ar is defined as an optionally substituted phenyl
group.
[0822] 1. N-Linkage Formation:
[0823] Methods A-E below describe the formation of the
N-Linkage.
[0824] Method A below describes an approach to amide formation.
[0825] Method A illustrates the construction of a amide
intermediate (A-3) through acylation of an aniline (A-2). The
acylating agent (A-1) comprises a leaving group (X). This leaving
group can be, for example, halogen, mesylate, acyl (as in an
anhydride), alcohol (as in ester/active ester) and the like. As
shown, the acylation process eliminates a molecule of HX.
[0826] A base can be used to facilitate the deprotonation of the
aniline and trapping of the HX byproduct. Suitable bases are
typically mild bases such as alkali carbonates (e.g.,
K.sub.2CO.sub.3).
##STR00215##
[0827] Method B shows the construction of a sulphonamide
intermediate (A-5) through the coupling of a sulphonyl halide (A-4)
with aniline (A-2).
##STR00216##
[0828] Method C shows the construction of a urea intermediate (A-7)
through the coupling of aniline (A-2) with an isocyanate (A-6)
##STR00217##
[0829] Method D shows the construction of an aniline intermediate
(A-8) through the reduction of amide (A-3) with the reducing agent
lithium aluminium hydride or the like.
##STR00218##
[0830] Method E shows the construction of an aniline intermediate
(A-8) through the Palladium catalysed cross-coupling of an aryl
halide (A-9) with an amine (A-10).
##STR00219##
[0831] 2. Side Chain Formation and Modification
[0832] Methods F-T describe methods for side chain formation and
modifications.
[0833] Generally, a suitably substituted phenyl derivative can be
coupled to a diverse range of side chains, which is further
modified to provide the final linkages and the nitrogen-containing
moieties of the compounds disclosed herein.
[0834] Methods F-T illustrate pathways to form propylene linkages
of the compounds disclosed herein.
[0835] Method F illustrates an aryl halide coupling with an allyl
alcohol in the presence of a palladium(0) catalyst. The terminal
alcohol group of allyl alcohol has been simultaneously oxidized to
an aldehyde group, which is further transformed to an amine via a
reductive amination.
##STR00220##
[0836] Method G illustrates a condensation between an aryl aldehyde
or aryl ketone and a nitrile having at least one .alpha.-hydrogen.
The resulting intermediate is further reduced to an amine.
##STR00221##
[0837] Method H is an acylation reaction to form a ketone-based
linkage. One skilled in the art will recognize that the R' group
may comprise functional groups that can be further modified.
##STR00222##
[0838] Method I is an ring-opening reaction of an epoxide to form a
hydroxy-substituted propylene side chain linkage.
##STR00223##
[0839] Method J is an attachment of side chain moieties via an
oxygen atom. More specifically, a side chain precursor (R'OH) can
be condensed with an aryl derivative by eliminating a molecule of
H.sub.2O. R' may comprise functional groups that can be further
modified to prepare linkages and nitrogen-containing moieties of
compounds disclosed herein.
##STR00224##
[0840] Method K is a condensation reaction that provides an oxygen
linking atom. Here, a molecule of HX is eliminated as the result of
the condensation.
##STR00225##
[0841] After attachment, the side chain moiety is optionally
further modified to provide the final linkage and the terminal
nitrogen-containing moiety for the compounds disclosed herein. The
following methods illustrate a variety of synthetic pathways to
modify the side chain moiety by reduction, oxidation, substitution,
fluorination, acylation and the like. Through application of these
methods, one of skill in the art recognizes that a diverse group of
linkages can be synthesized.
[0842] Method L illustrates an amination process in which
carboxylic acid is converted to an amine. Typically, the carboxylic
acid (or ester) can be first reduced to primary alcohol, which can
then be converted to an amine via mesylate, halide, azide,
phthalimide, or Mitsunobu reaction and the like. Suitable reducing
agents include, for example, lithium aluminum hydride (LiAlH.sub.4)
and the like. As shown, the resulting amine can be further
functionalized, by known methods in the art.
##STR00226##
[0843] Additional or alternative modifications can be carried out
according to the methods illustrated below.
##STR00227##
[0844] As a non-limiting example only, Scheme A illustrates a
complete synthetic sequence for preparing a compound disclosed
herein.
##STR00228##
[0845] In Scheme A, the three carbon side chain is introduced
through alkylation of 3-nitrobenzaldehyde with acetonitrile. The
sulphonamide is introduced by a two step process involving
reduction of the nitro group followed by reaction with a sulphonyl
halide. Finally, reduction of the nitrile to an amine gives the
target compound.
Methods for Preparing Compounds of Formula (II)
[0846] Generally speaking, the compounds disclosed herein can be
prepared in a stepwise manner involving an olefin formation and a
side chain formation.
[0847] In certain embodiments, an olefin intermediate can be first
constructed, which forms the precursor to the styrenyl core
structure. A side chain moiety, which is a precursor to the linkage
and the nitrogen-containing moiety of the compounds disclosed
herein, can then be attached to the olefin intermediate.
[0848] In other embodiments, the compounds disclosed herein can be
prepared by first preparing a phenyl intermediate having an
appropriate side chain, followed by an olefin formation to provide
the styrenyl core structure.
[0849] The following Methods illustrate various synthetic pathways
for preparing olefin intermediates and the side chain moieties. One
skilled in the art will recognize that a method for olefin
formation can be combined with a method for side chain formation to
provide the compounds disclosed herein. For example, Method A can
be combined with any of Method K, Methods K and U, Methods K and L,
Methods K and AB, Methods T and L, Method R, Method S, Method J,
Method E, Methods R and U, and the like. Similarly, Method C can be
combined with Method J.
Olefin Formation:
[0850] Methods A-I below describe various approaches to olefin
formation.
[0851] More specifically, Method A illustrates constructing an
olefin intermediate (A-3) in a
[0852] Wittig reaction. Depending on the sequence of the reactions,
Ar can be a phenyl derivative compound that is already attached to
a side chain moiety, or Ar may comprise a reactive group
(appropriately protected), which will be coupled to a side chain
moiety after the olefin formation step.
[0853] According to Method A, a phosphonium ylide reagent (or
"Wittig reagent") (A-1) can be coupled to a benzaldehyde or ketone
derivative (A-2) to provide the olefin intermediate (A-3) in the
presence of a base. The geometry of the resulting A-3 may depend on
the reactivity of the ylide reagent. Triphenylphosphonium-based
ylide reagent (R is phenyl) typically produces predominantly (E) or
trans-styrenes; whereas trialkylphosphonium-based ylide reagent (R
is alkyl) produces predominantly (Z) or cis-styrene. The E or Z
stereoisomers can be separated by, for example, chromatography or
other known methods in the art.
[0854] The ylide reagent (A-1) can be prepared according to known
methods in the art. For example, R.sub.11--CH.sub.2OH can be
converted to the corresponding ylide reagent (A-1) in the presence
of triphenylphosphine hydrobromide. The benzaldehyde or ketone
derivative (A-2) may be commercially available or can be prepared
by known methods in the art.
[0855] The olefin intermediate (A-3) may also be prepared by
coupling a phosphonium ylide reagent derivatized from the Ar group
(A-4) and an aldehyde or ketene derivative of R.sub.11 (A-5). The
ylide reagent (A-4) can be prepared from, for example, a benzyl
alcohol, whereas (A-5) can be prepared by known methods in the art
or can be obtained from commercial vendors.
##STR00229##
[0856] Method AE shows a coupling reaction similar to the Wittig
reaction of Method A, except that a phosphorus ylide is used in
place of the phosphonium ylide. The phosphorus ylide can be coupled
to an aldehyde or ketone in the presence of a base
(Wittig-Horner-Emmons reaction.)
##STR00230##
[0857] In addition, elimination reactions can be used to form
olefin bonds. Methods B-D illustrate various approaches to forming
alcohol precursors that can undergo alcohol dehydration in acidic
conditions to produce olefin bonds. The Ar group is typically
activated with a metal (e.g., Li) to facilitate the alcohol
formation. Grignard reagent can also be used in place of the
metal.
[0858] As discussed above in connection with Method A, the alcohol
precursor in each of Methods B-D can also be prepared by using a
metal activated R.sub.11 group and an Ar group derivatized with a
carbonyl group or a cyclopropyl group.
##STR00231##
[0859] Methods E-G illustrate coupling an olefin or an activated
olefin directly with an aryl halide in the presence of a
palladium(0) catalyst. In certain embodiments, the olefin can be
activated by a transition metal (e.g., Zn or Sn), or boronic acid
(e.g., Suzuki reaction), as are known in the art. The halo
substituent of the aryl group can be, for example, bromo or
iodo.
[0860] Palladium catalysts suitable for coupling reactions are
known to one skilled in the art.
[0861] Exemplary palladium(0) catalysts include, for example,
tetrakis(triphenylphosphine)palladium(0) [Pd(PPh.sub.3).sub.4] and
tetrakis(tri(o-tolylphosphine)palladium(0),
tetrakis(dimethylphenylphosphine)palladium(0),
tetrakis(tris-p-methoxyphenylphosphine)palladium(0) and the like.
It is understood that a palladium (II) salt can also be used, which
generates the palladium (0) catalyst in situ. Suitable palladium
(II) salts include, for example, palladium diacetate
[Pd(OAc).sub.2], bis(triphenylphosphine)-palladium diacetate and
the like.
##STR00232##
[0862] An olefin intermediate can also be constructed from an
alkyne addition/hydrogenation reaction. Depending on the reaction
conditions (syn or anti addition), cis or trans configuration can
be formed.
[0863] Method H illustrates a syn-addition, i.e., both hydrogens
are added from one side of the alkyne molecule, which results in a
cis olefin configuration. Typically, hydrogen gas can be used in
the presence of a catalyst (e.g., Pd on carbon or platinum) to
effect a syn addition.
##STR00233##
[0864] Method I illustrate an anti-addition, i.e., an adding agent
is added to opposite sides of the alkyne molecule, resulting in a
trans olefin configuration. The adding agent can be, for example,
aluminum hydride reagents, lithium/NH.sub.3 reagents and the
like.
##STR00234##
Side Chain Formation and Modification
[0865] Methods J-T and AA-AD below describe various approaches to
side chain formation and modifications.
[0866] Generally speaking, a suitably substituted phenyl derivative
can be coupled to a diverse range of side chains, which may be
further modified to provide the final linkages and the
nitrogen-containing moieties of the compounds disclosed herein.
[0867] Method J illustrates an aryl halide coupled with an allyl
alcohol in the presence of a palladium(0) catalyst. The terminal
alcohol group of allyl alcohol has been simultaneously oxidized to
an aldehyde group, which can be further reduced to an amine
(--NR.sub.9R.sub.10).
##STR00235##
[0868] Method K illustrates an aldol condensation between an aryl
aldehyde or aryl ketone with a nitrile reagent comprising at least
one .alpha.-hydrogen. The resulting condensation intermediate can
be further reduced to an amine (--NR.sub.9R.sub.10).
##STR00236##
[0869] Method AA shows an acylation reaction to form a ketone-based
linkage. One skilled in the art will recognize that the R' group
may comprise functional groups that can be further modified.
##STR00237##
[0870] Method R shows a ring-opening reaction of an epoxide reagent
to form a 3-carbon side chain linkage.
##STR00238##
[0871] Method S shows the formation of a triple bond linkage based
on a Sonogashira reaction. Typically, palladium(0) catalyst is used
in combination with a base to couple an aryl halide with a
acetylene derivative. R' can be further modified, as described
herein.
##STR00239##
[0872] Method T shows the formation of a double bond linkage based
on a Heck reaction. Typically, palladium(0) catalyst is used in
combination with a base to couple an aryl halide with a vinyl
derivative. R' can be further modified, as described herein.
##STR00240##
[0873] Methods M-P illustrate attachments of side chain moieties by
heteroatoms. Method M shows a side chain precursor (R'OH) attached
to an aryl derivative via an oxygen atom in a condensation reaction
in which a molecule of H.sub.2O is eliminated. R' may comprise
functional groups that can be further modified to prepare linkages
and nitrogen-containing moieties of the compounds disclosed
herein.
##STR00241##
[0874] Method N shows a similar coupling reaction that provides a
sulfur linking atom. Method O illustrates an oxidation step of the
sulfur linking atom to provide --S(O)-- or --S(O).sub.2--,
depending on the degree of oxidation.
##STR00242##
[0875] Method P shows the formation of an amide-containing linkage,
in which a aniline derivative is coupled with a carboxylic acid
derivative. The carboxylic acid derivative can be activated to
facilitate the amide formation. Suitable activating reagents
include, for example, 1,3-dicyclohexylcarbodiimide (DCC),
1,1'-carbonyldiimidazole (CDI),
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCL),
benzotriazol-1-yl-oxy-tris(dimethylamino)phosphonium
hexafluorophosphate (BOP), and 1,3-diisopropylcarbodiimide
(DICD).
##STR00243##
[0876] After attachment, the side chain moiety can be further
modified to provide the final linkage and the terminal
nitrogen-containing moiety for the compounds disclosed herein. The
following methods illustrate a variety of synthetic pathways to
manipulate or modify the side chain moiety by reduction, oxidation,
nucleophilic or electrophilic substitution, fluorination, acylation
and the like. As a result, a diverse group of linkages can be
synthesized.
[0877] Method L illustrates an amination process in which
carboxylic acid is converted to an amine. Typically, the carboxylic
acid (or ester) can be first reduced to primary alcohol, which can
then be converted to an amine via mesylate, halide, azide,
phthalimide, or Mitsunobu reaction and the like. Suitable reducing
agents include, for example, sodium borohydride (NaBH.sub.4),
sodium cyanoborohydride (NaBH.sub.3CN), sodium
triacetoxyborohydride (NaBH(OCOCH.sub.3).sub.3), lithium aluminum
hydride (LiAlH.sub.4) and the like. As shown, the resulting amine
can be further functionalized, by known methods in the art.
##STR00244##
[0878] Additional or alternative modifications can be carried out
according to the methods illustrated below.
##STR00245## ##STR00246##
[0879] Scheme I illustrates a complete synthetic sequence for
preparing one example of the compounds disclosed herein.
##STR00247##
[0880] In Scheme I, an olefin intermediate is first constructed,
followed by coupling to a side chain moiety. Further modification
of the side chain moiety by reduction affords the compounds
disclosed herein having a propylene linkage and a terminal amine.
Other nitrogen-containing moieties can be further derived from the
terminal amine, according to known methods in the art.
[0881] One skilled in the art should recognize, however, that the
order of the reactions may vary. Thus, in other embodiments, as
shown in Scheme II, a side chain attachment is initially performed,
followed by olefin formation.
##STR00248##
[0882] Additional methods for preparing compounds of Formula (II)
are disclosed in WO 2008/131368, which is incorporated by reference
in its entirety.
Methods for Preparing Compounds of Formula (III)
[0883] Generally speaking, compounds disclosed herein can be
prepared in a stepwise manner involving an acetylene formation and
a side chain formation of a phenyl ring. Typically, the acetylene
formation can take place by attaching an acetylene precursor to a
phenyl. For example, in certain embodiments, an acetylene
intermediate can be first constructed, which forms the precursor to
the alkynyl phenyl core structure. A side chain moiety, which is a
precursor to the linkage (i.e., propylene or ethylene oxide) and
the nitrogen-containing moiety of the compounds disclosed herein,
can then be attached to the acetylene intermediate.
[0884] In other embodiments, the compounds disclosed herein can be
prepared by first preparing a phenyl intermediate having an
appropriate side chain, followed by an acetylene formation to
provide the alkynyl core structure.
[0885] The following Methods illustrate various synthetic pathways
for preparing acetylene intermediates and the side chain moieties.
One skilled in the art will recognize that a method for acetylene
formation can be combined with a method for side chain formation to
provide the compounds disclosed herein. For example, any one of
Methods A-D can be combined with any of Methods E-H, or any of
Methods I-J. They can be further combined with any of Methods K-S
to modify the linkage and/or the terminal nitrogen-containing
moiety.
Acetylene Formation:
[0886] Methods A-D below describe various approaches to acetylene
formation.
[0887] More specifically, Method A illustrates the construction of
an acetylene intermediate (A-3) in a Sonogashira or Castro-Stephens
reaction. Depending on the sequence of the reactions, Ar can be a
phenyl derivative compound that is already attached to a side chain
moiety, or Ar may comprise a reactive group (appropriately
protected), which will be coupled to a side chain moiety after the
acetylene formation step.
[0888] According to Method A, an alkyne (A-1) can be coupled to an
aryl halide or a reactive equivalent (A-2) to provide the acetylene
intermediate (A-3) in the presence of a copper (I) catalyst
(Castro-Stephens) or a mixture of Pd.sup.0 and Cu.sup.1 catalysts
(Sonogashira).
[0889] The alkyne (A-1) has a terminal acetylene structure that is
capable of coupling to A-2. Alkynes comprising diverse R.sub.5
groups can be prepared according to known methods in the art. For
example, organic halides (e.g., R.sub.5Br) can be converted to the
corresponding alkyne (A-1) by coupling to an ethyne. The
halobenzene or its reactive equivalent (A-2) may be commercially
available or can be prepared by known methods in the art.
[0890] Palladium catalysts suitable for coupling reactions are
known to one skilled in the art. Exemplary palladium(0) catalysts
include, for example, tetrakis(triphenylphosphine)palladium(0)
[Pd(PPh.sub.3).sub.4] and
tetrakis(tri(o-tolylphosphine)palladium(0),
tetrakis(dimethylphenylphosphine)palladium(0),
tetrakis(tris-p-methoxyphenylphosphine)palladium(0) and the like.
It is understood that a palladium (II) salt can also be used, which
generates the palladium (0) catalyst in situ. Suitable palladium
(II) salts include, for example, palladium diacetate
[Pd(OAc).sub.2], bis(triphenylphosphine)-palladium diacetate and
the like.
[0891] Copper catalysts suitable for coupling reactions are known
to one skilled in the art. Typically, the copper (I) catalyst can
be copper (I) iodide.
##STR00249##
[0892] Method B shows an alternative construction of the acetylene
intermediate (A-3) by coupling an organic halide (i.e., R.sub.5X)
with a phenyl comprising a terminal acetylene (A-5).
##STR00250##
[0893] Method C shows the construction of an acetylene intermediate
(A-7) through the addition of a terminal acetylene (A-5) to an
aldehyde or ketone (A-6).
##STR00251##
[0894] Method D shows the construction of an acetylene intermediate
(A-8) through the addition of a terminal acetylene (A-5) to an
epoxide (A-9).
##STR00252##
Side Chain Formation and Modification
[0895] Methods E-S below describe various approaches to side chain
formation and modifications.
[0896] Generally speaking, a suitably substituted phenyl derivative
can be coupled to a diverse range of side chains, which may be
further modified to provide the final linkages and the
nitrogen-containing moieties of the compounds disclosed herein.
[0897] Methods E-H illustrate pathways to form propylene linkages
of the compounds disclosed herein.
[0898] Method E illustrates an aryl halide coupled with an allyl
alcohol in the presence of a palladium(0) catalyst. The terminal
alcohol group of allyl alcohol has been simultaneously oxidized to
an aldehyde group, which can be further reductively aminated to an
amine (--NR.sub.12R.sub.13).
##STR00253##
[0899] Method F illustrates an aldol condensation between an aryl
aldehyde or aryl ketone with a nitrile reagent comprising at least
one .alpha.-hydrogen. The resulting condensation intermediate can
be further reduced to an amine (--NR.sub.12R.sub.13).
##STR00254##
[0900] Method G shows an acylation reaction to form a ketone-based
linkage (i.e., R.sub.10 and R.sub.11 of Formula (I) form an oxo).
One skilled in the art will recognize that the R' group may
comprise functional groups that can be further modified.
##STR00255##
[0901] Method H shows a ring-opening reaction of an epoxide reagent
to form a hydroxy-substituted propylene side chain linkage.
##STR00256##
[0902] Method I illustrates an attachment of side chain moieties by
an oxygen, which can be a precursor to an ethylene oxide linkage.
More specifically, a side chain precursor (R'OH) can be condensed
with an aryl derivative by eliminating a molecule of H.sub.2O. R'
may comprise functional groups that can be further modified to
prepare linkages and nitrogen-containing moieties of compounds of
Formula (III) and its substructures, including Formulae (IIIa) and
(IIIb).
##STR00257##
[0903] Method J shows a condensation reaction that provides an
oxygen linking atom. Here, a molecule of HX is eliminated as the
result of the condensation.
##STR00258##
[0904] After attachment, the side chain moiety can be further
modified to provide the final linkage and the terminal
nitrogen-containing moiety for the compounds disclosed herein. The
following methods illustrate a variety of synthetic pathways to
manipulate or modify the side chain moiety by reduction, oxidation,
nucleophilic or electrophilic substitution, fluorination, acylation
and the like. As a result, a diverse group of linkages can be
synthesized.
[0905] Method K illustrates an amination process in which
carboxylic acid is converted to an amine. Typically, the carboxylic
acid (or ester) can be first reduced to primary alcohol, which can
then be converted to an amine via mesylate, halide, azide,
phthalimide, or Mitsunobu reaction and the like. Suitable reducing
agents include, for example, sodium borohydride (NaBH.sub.4),
sodium cyanoborohydride (NaBH.sub.3CN), sodium
triacetoxyborohydride (NaBH(OCOCH.sub.3).sub.3), lithium aluminum
hydride (LiAlH.sub.4) and the like. As shown, the resulting amine
can be further functionalized, by known methods in the art.
##STR00259##
[0906] Additional or alternative modifications can be carried out
according to the methods illustrated below.
##STR00260## ##STR00261## ##STR00262## ##STR00263## ##STR00264##
##STR00265## ##STR00266## ##STR00267##
[0907] Scheme I illustrates a complete synthetic sequence for
preparing a compound disclosed herein.
##STR00268##
[0908] In Scheme I, the side chain moiety is first constructed and
the amine protected. The acetylene moiety is then formed through
coupling with a terminal acetylene according to Method A. The
coupling product is then deprotected to give rise to the final
alkynyl phenyl derivative compound comprising a propylene linkage
terminating in a primary amine Other nitrogen-containing moieties
(--NR.sub.12R.sub.13) can be further derived from the terminal
amine, according to known methods in the art.
[0909] One skilled in the art should recognize, however, that the
order of the reactions may vary. Thus, in other embodiments,
acetylene formation may precede the side chain attachment.
[0910] Scheme II illustrates a complete synthetic sequence for
preparing a compound disclosed herein.
##STR00269##
[0911] Additional methods for preparing compounds of Formula (III)
are disclosed in WO 2009/005794, which is incorporated by reference
in its entirety.
Methods for Preparing Compounds of Formula (IV)
[0912] Compounds disclosed herein can be prepared in a stepwise
manner involving alkylation of a phenol and construction of the
linker to the amine
Alkylation:
[0913] Methods A-B below describe various approaches to
alkylation.
[0914] More specifically, Method A illustrates the construction of
an alkoxy intermediate (A-3) through alkylation of a phenol (A-2).
The alkylating agent (A-1) comprises a moiety (X) reactive to the
acidic hydrogen of phenol. X can be, for example, halogen,
mesylate, tosylate, triflate and the like. As shown, the alkylation
process eliminates a molecule of HX.
[0915] A base can be used to facilitate the deprotonation of the
phenol. Suitable bases are typically mild bases such as alkali
carbonates (e.g., K.sub.2CO.sub.3). Depending on X, other reagents
(e.g., PPh.sub.3 in combination with DEAD) can be used to
facilitate the alkylation process.
##STR00270##
[0916] Method B shows the construction of an alkoxy intermediate
(A-5) through the ring-opening of an epoxide (A-4).
##STR00271##
Side Chain Formation and Modification
[0917] Methods C-P below describe various approaches to side chain
formation and modifications.
[0918] Generally speaking, a suitably substituted aryl derivative
(e.g., alkoxyphenyl) can be coupled to a diverse range of side
chains, which may be further modified to provide the final linkages
and the nitrogen-containing moieties of compounds disclosed
herein.
[0919] Method C illustrates an aldol condensation between an aryl
aldehyde or aryl ketone with a nitrile reagent comprising at least
one .alpha.-hydrogen. The resulting condensation intermediate can
be further reduced to an amine (--NH.sub.2).
##STR00272##
[0920] Method D shows an acylation reaction to form a ketone-based
linkage. One skilled in the art will recognize that the R' group
comprises functional groups that can be further modified.
##STR00273##
[0921] Method E shows a ring-opening reaction of an epoxide reagent
to form a 3-carbon side chain linkage. R' can be further
modified.
##STR00274##
[0922] Method F shows the formation of a triple bond linkage based
on a Sonogashira reaction.
[0923] Typically, palladium(0) catalyst is used in combination with
a base to couple an aryl halide with a acetylene derivative. R' can
be further modified, as described herein. The acetylene linkage can
also be further modified, for example, by hydrogenation to provide
alkylene or alkenylene linkage.
##STR00275##
[0924] Palladium catalysts suitable for coupling reactions are
known to one skilled in the art.
[0925] Exemplary palladium(0) catalysts include, for example,
tetrakis(triphenylphosphine)palladium(0) [Pd(PPh.sub.3).sub.4] and
tetrakis(tri(o-tolylphosphine)palladium(0),
tetrakis(dimethylphenylphosphine)palladium(0),
tetrakis(tris-p-methoxyphenylphosphine)palladium(0) and the like.
It is understood that a palladium (II) salt can also be used, which
generates the palladium (0) catalyst in situ. Suitable palladium
(II) salts include, for example, palladium diacetate
[Pd(OAc).sub.2], bis(triphenylphosphine)-palladium diacetate and
the like.
[0926] Method G shows the formation of a double bond linkage based
on a Heck reaction. Typically, palladium(0) catalyst is used in
combination with a base to couple an aryl halide with a vinyl
derivative. R' can be further modified, as described herein.
##STR00276##
[0927] Methods H-P illustrate attachments of side chain moieties by
heteroatoms. Method H shows a side chain precursor (R'OH) attached
to an aryl derivative via an oxygen atom in a condensation reaction
in which a molecule of water is eliminated. R' comprises functional
groups that can be further modified to prepare linkages and
nitrogen-containing moieties of the compounds disclosed herein.
##STR00277##
[0928] Additional or alternative modifications can be carried out
according to the methods illustrated below.
##STR00278## ##STR00279## ##STR00280## ##STR00281## ##STR00282##
##STR00283## ##STR00284## ##STR00285##
[0929] Scheme I illustrates a complete synthetic sequence for
preparing a compound disclosed herein.
##STR00286##
[0930] In Scheme I, the alkoxy intermediate is formed via
alkylation of a phenol. The side chain is introduced through a
Sonogashira coupling. Deprotection of the amine, followed by
hydrogenation of the acetylene gives the target compound. Other
nitrogen-containing moieties can be further derived from the
terminal amine, according to known methods in the art.
[0931] Additional methods for preparing compounds of Formula (IV)
are disclosed in WO 2009/045479, which is incorporated by reference
in its entirety.
[0932] In addition to the generic reaction schemes and methods
discussed above, other exemplary reaction schemes are also provided
to illustrate methods for preparing compounds described herein or
any of its subgenus structures.
II. Treatment of Ophthalmic Diseases and Disorders
[0933] Compounds as described herein, including compounds having
the structure as set forth in Formula (I), (II), (IIa), (III),
(IIIa), (IV), or (IVa) and substructures thereof, are useful for
treating an ophthalmic disease or disorder by inhibiting one or
more steps in the visual cycle. In some embodiments, the compounds
disclosed herein function by inhibiting or blocking the activity of
a visual cycle trans-cis isomerase. The compounds described herein,
may inhibit, block, or in some manner interfere with the
isomerization step in the visual cycle. In a particular embodiment,
the compound inhibits isomerization of an all-trans-retinyl ester;
in certain embodiments, the all-trans-retinol ester is a fatty acid
ester of all-trans-retinol, and the compound inhibits isomerization
of all-trans-retinol to 11-cis-retinol. The compound may bind to,
or in some manner interact with, and inhibit the isomerase activity
of at least one visual cycle isomerase, which may also be referred
to herein and in the art as a retinal isomerase or an
isomerohydrolase. The compound may block or inhibit binding of an
all-trans-retinol ester substrate to an isomerase. Alternatively,
or in addition, the compound may bind to the catalytic site or
region of the isomerase, thereby inhibiting the capability of the
enzyme to catalyze isomerization of an all-trans-retinol ester
substrate. On the basis of scientific data to date, an at least one
isomerase that catalyzes the isomerization of all-trans-retinyl
esters is believed to be located in the cytoplasm of RPE cells. As
discussed herein, each step, enzyme, substrate, intermediate, and
product of the visual cycle is not yet elucidated (see, e.g.,
Moiseyev et al., Proc. Natl. Acad. Sci. USA 102:12413-18 (2004);
Chen et al., Invest. Ophthalmol. Vis. Sci. 47:1177-84 (2006); Lamb
et al. supra).
[0934] Compounds of Formula (II), (IIa), (III), (IIIa), (IV) or
(IVa) as described herein, and substructures thereof, are useful
for treating an ophthalmic disease or disorder by inhibiting one or
more steps in the visual cycle. The compounds described herein may
be useful for treating a subject who has an ophthalmic disease or
disorder, particularly a retinal disease or disorder such as
age-related macular degeneration or Stargardt's macular
dystrophy.
[0935] A method for determining the effect of a compound on
isomerase activity may be performed in vitro as described herein
and in the art (Stecher et al., J Biol Chem 274:8577-85 (1999); see
also Golczak et al., Proc. Natl. Acad. Sci. USA 102:8162-67
(2005)). Retinal pigment epithelium (RPE) microsome membranes
isolated from an animal (such as bovine, porcine, human, for
example) may serve as the source of the isomerase. The capability
of the compounds described herein to inhibit isomerase may also be
determined by an in vivo murine isomerase assay. Brief exposure of
the eye to intense light ("photobleaching" of the visual pigment or
simply "bleaching") is known to photo-isomerize almost all
11-cis-retinal in the retina. The recovery of 11-cis-retinal after
bleaching can be used to estimate the activity of isomerase in vivo
(see, e.g., Maeda et al., J. Neurochem 85:944-956 (2003); Van
Hooser et al., J Biol Chem 277:19173-82, 2002).
Electroretinographic (ERG) recording may be performed as previously
described (Haeseleer et al., Nat. Neurosci. 7:1079-87 (2004);
Sugitomo et al., J. Toxicol. Sci. 22 Suppl 2:315-25 (1997); Keating
et al., Documenta Ophthalmologica 100:77-92 (2000)). See also
Deigner et al., Science, 244: 968-971 (1989); Gollapalli et al.,
Biochim Biophys Acta. 1651: 93-101 (2003); Parish, et al., Proc.
Natl. Acad. Sci. USA 95:14609-13 (1998); Radu, et al., Proc Natl
Acad Sci USA 101: 5928-33 (2004)). In certain embodiments,
compounds that are useful for treating a subject who has or who is
at risk of developing any one of the ophthalmic and retinal
diseases or disorders described herein have IC.sub.50 levels
(compound concentration at which 50% of isomerase activity is
inhibited) as measured in the isomerase assays described herein or
known in the art that is less than about 1 .mu.M; in other
embodiments, the determined IC.sub.50 level is less than about 10
nM; in other embodiments, the determined IC.sub.50 level is less
than about 50 nM; in certain other embodiments, the determined
IC.sub.50 level is less than about 100 nM; in other certain
embodiments, the determined IC.sub.50 level is less than about 10
.mu.M; in other embodiments, the determined IC.sub.50 level is less
than about 50 .mu.M; in other certain embodiments, the determined
IC.sub.50 level is less than about 100 .mu.M or about 500 .mu.M; in
other embodiments, the determined IC.sub.50 level is between about
1 .mu.M and 10 .mu.M; in other embodiments, the determined
IC.sub.50 level is between about 1 nM and 10 nM. When adminstered
into a subject, one or more compounds of the present invention
exhibits an ED.sub.50 value of about 5 mg/kg, 5 mg/kg or less as
ascertained by inhibition of an isomerase reaction that results in
production of 11-cis-retinol. In some embodiments, the compounds of
the present invention have ED.sub.50 values of about 1 mg/kg when
administered into a subject. In other embodiments, the compounds of
the present invention have ED.sub.50 values of about 0.1 mg/kg when
administered into a subject. The ED.sub.50 values can be measured
after about 2 hours, 4 hours, 6 hours, 8 hours or longer upon
administering a subject compound or a pharmaceutical composition
thereof.
[0936] The compounds described herein may be useful for treating a
subject who has an ophthalmic disease or disorder, particularly a
retinal disease or disorder such as age-related macular
degeneration or Stargardt's macular dystrophy. In one embodiment,
the compounds described herein may inhibit (i.e., prevent, reduce,
slow, abrogate, or minimize) accumulation of lipofuscin pigments
and lipofuscin-related and/or associated molecules in the eye. In
another embodiment, the compounds may inhibit (i.e., prevent,
reduce, slow, abrogate, or minimize)
N-retinylidene-N-retinylethanolamine (A2E) accumulation in the eye.
The ophthalmic disease may result, at least in part, from
lipofuscin pigments accumulation and/or from accumulation of A2E in
the eye. Accordingly, in certain embodiments, methods are provided
for inhibiting or preventing accumulation of lipofuscin pigments
and/or A2E in the eye of a subject. These methods comprise
administering to the subject a composition comprising a
pharmaceutically acceptable or suitable excipient (i.e.,
pharmaceutically acceptable or suitable carrier) and a compound as
described in detail herein, including a compound having the
structure as set forth in Formula (I), (II), (IIa), (III), (IIIa),
(IV), or (IVa) and substructures thereof, and the specific
compounds described herein.
[0937] Accumulation of lipofuscin pigments in retinal pigment
epithelium (RPE) cells has been linked to progression of retinal
diseases that result in blindness, including age-related macular
degeneration (De Laey et al., Retina 15:399-406 (1995)). Lipofuscin
granules are autofluorescent lysosomal residual bodies (also called
age pigments). The major fluorescent species of lipofuscin is A2E
(an orange-emitting fluorophore), which is a positively charged
Schiff-base condensation-product formed by all-trans retinaldehyde
with phosphatidylethanolamine (2:1 ratio) (see, e.g., Eldred et
al., Nature 361:724-6 (1993); see also, Sparrow, Proc. Natl. Acad.
Sci. USA 100:4353-54 (2003)). Much of the indigestible lipofuscin
pigment is believed to originate in photoreceptor cells; deposition
in the RPE occurs because the RPE internalize membranous debris
that is discarded daily by the photoreceptor cells. Formation of
this compound is not believed to occur by catalysis by any enzyme,
but rather A2E forms by a spontaneous cyclization reaction. In
addition, A2E has a pyridinium bisretinoid structure that once
formed may not be enzymatically degraded. Lipofuscin, and thus A2E,
accumulate with aging of the human eye and also accumulate in a
juvenile form of macular degeneration called Stargardt's disease,
and in several other congenital retinal dystrophies.
[0938] A2E may induce damage to the retina via several different
mechanisms. At low concentrations, A2E inhibits normal proteolysis
in lysosomes (Holz et al., Invest. Ophthalmol. Vis. Sci. 40:737-43
(1999)). At higher, sufficient concentrations, A2E may act as a
positively charged lysosomotropic detergent, dissolving cellular
membranes, and may alter lysosomal function, release proapoptotic
proteins from mitochondria, and ultimately kill the RPE cell (see,
e.g., Eldred et al., supra; Sparrow et al., Invest. Ophthalmol.
Vis. Sci. 40:2988-95 (1999); Holz et al., supra; Finneman et al.,
Proc. Natl. Acad. Sci. USA 99:3842-347 (2002); Suter et al., J.
Biol. Chem. 275:39625-30 (2000)). A2E is phototoxic and initiates
blue light-induced apoptosis in RPE cells (see, e.g., Sparrow et
al., Invest. Ophthalmol. Vis. Sci. 43:1222-27 (2002)). Upon
exposure to blue light, photooxidative products of A2E are formed
(e.g., epoxides) that damage cellular macromolecules, including DNA
(Sparrow et al., J. Biol. Chem. 278(20):18207-13 (2003)). A2E
self-generates singlet oxygen that reacts with A2E to generate
epoxides at carbon-carbon double bonds (Sparrow et al., supra).
Generation of oxygen reactive species upon photoexcitation of A2E
causes oxidative damage to the cell, often resulting in cell death.
An indirect method of blocking formation of A2E by inhibiting
biosynthesis of the direct precursor of A2E, all-trans-retinal, has
been described (see U.S. Patent Application Publication No.
2003/0032078). However, the usefulness of the method described
therein is limited because generation of all-trans retinal is an
important component of the visual cycle. Other therapies described
include neutralizing damage caused by oxidative radical species by
using superoxide-dismutase mimetics (see, e.g., U.S. Patent
Application Publication No. 2004/0116403) and inhibiting
A2E-induced cytochrome C oxidase in retinal cells with negatively
charged phospholipids (see, e.g., U.S. Patent Application
Publication No. 2003/0050283).
[0939] The compounds described herein may be useful for preventing,
reducing, inhibiting, or decreasing accumulation (i.e., deposition)
of A2E and A2E-related and/or derived molecules in the RPE. Without
wishing to be bound by theory, because the RPE is critical for the
maintenance of the integrity of photoreceptor cells, preventing,
reducing, or inhibiting damage to the RPE may inhibit degeneration
(i.e., enhance the survival or increase or prolong cell viability)
of retinal neuronal cells, particularly, photoreceptor cells.
Compounds that bind specifically to or interact with A2E
A2E-related and/or derived molecules or that affect A2E formation
or accumulation may also reduce, inhibit, prevent, or decrease one
or more toxic effects of A2E or of A2E-related and/or derived
molecules that result in retinal neuronal cell (including a
photoreceptor cell) damage, loss, or neurodegeneration, or in some
manner decrease retinal neuronal cell viability. Such toxic effects
include induction of apoptosis, self-generation of singlet oxygen
and generation of oxygen reactive species; self-generation of
singlet oxygen to form A2E-epoxides that induce DNA lesions, thus
damaging cellular DNA and inducing cellular damage; dissolving
cellular membranes; altering lysosomal function; and effecting
release of proapoptotic proteins from mitochondria.
[0940] In other embodiments, the compounds described herein may be
used for treating other ophthalmic diseases or disorders, for
example, glaucoma, cone-rod dystrophy, retinal detachment,
hemorrhagic or hypertensive retinopathy, retinitis pigmentosa,
optic neuropathy, inflammatory retinal disease, proliferative
vitreoretinopathy, genetic retinal dystrophies, traumatic injury to
the optic nerve (such as by physical injury, excessive light
exposure, or laser light), hereditary optic neuropathy, neuropathy
due to a toxic agent or caused by adverse drug reactions or vitamin
deficiency, Sorsby's fundus dystrophy, uveitis, a retinal disorder
associated with Alzheimer's disease, a retinal disorder associated
with multiple sclerosis; a retinal disorder associated with viral
infection (cytomegalovirus or herpes simplex virus), a retinal
disorder associated with Parkinson's disease, a retinal disorder
associated with AIDS, or other forms of progressive retinal atrophy
or degeneration. In another specific embodiment, the disease or
disorder results from mechanical injury, chemical or drug-induced
injury, thermal injury, radiation injury, light injury, laser
injury. The subject compounds are useful for treating both
hereditary and non-hereditary retinal dystrophy. These methods are
also useful for preventing ophthalmic injury from environmental
factors such as light-induced oxidative retinal damage,
laser-induced retinal damage, "flash bomb injury," or "light
dazzle", refractive errors including but not limited to myopia
(see, e.g., Quinn G E et al. Nature 1999; 399:113-114; Zadnik K et
al. Nature 2000; 404:143-144; Gwiazda J et al. Nature 2000; 404:
144), etc.
[0941] In other embodiments, methods are provided herein for
inhibiting neovascularization (including but not limited to
neovascular glycoma) in the retina using any one or more of the
compounds as described in detail herein, including a compound
having the structure as set forth in Formula (I), (II), (IIa),
(III), (IIIa), (IV), or (IVa) and substructures thereof, and the
specific compounds described herein. In certain other embodiments,
methods are provided for reducing hypoxia in the retina using the
compounds described herein. These methods comprise administering to
a subject, in need thereof, a composition comprising a
pharmaceutically acceptable or suitable excipient (i.e.,
pharmaceutically acceptable or suitable carrier) and a compound as
described in detail herein, including a compound having the
structure as set forth in Formula (I), (II), (IIa), (III), (IIIa),
(IV), or (IVa) and substructures thereof, and the specific
compounds described herein.
[0942] Merely by way of explanation and without being bound by any
theory, and as discussed in further detail herein, dark-adapted rod
photoreceptors engender a very high metabolic demand (i.e.,
expenditure of energy (ATP consumption) and consumption of oxygen).
The resultant hypoxia may cause and/or exacerbate retinal
degeneration, which is likely exaggerated under conditions in which
the retinal vasculature is already compromised, including, but not
limited to, such conditions as diabetic retinopathy, macular edema,
diabetic maculopathy, retinal blood vessel occlusion (which
includes retinal venous occlusion and retinal arterial occlusion),
retinopathy of prematurity, ischemia reperfusion related retinal
injury, as well as in the wet form of age-related macular
degeneration (AMD). Furthermore, retinal degeneration and hypoxia
may lead to neovascularization, which in turn may worsen the extent
of retinal degeneration. The compounds described herein that
modulate the visual cycle can be administered to prevent, inhibit,
and/or delay dark adaptation of rod photoreceptor cells, and may
therefore reduce metabolic demand, thereby reducing hypoxia and
inhibiting neovascularization.
[0943] By way of background, oxygen is a critical molecule for
preservation of retinal function in mammals, and retinal hypoxia
may be a factor in many retinal diseases and disorders that have
ischemia as a component. In most mammals (including humans) with
dual vascular supply to the retina, oxygenation of the inner retina
is achieved through the intraretinal microvasculature, which is
sparse compared to the choriocapillaris that supplies oxygen to the
RPE and photoreceptors. The different vascular supply networks
create an uneven oxygen tension across the thickness of the retina
(Cringle et al., Invest. Ophthalmol. Vis. Sci. 43:1922-27 (2002)).
Oxygen fluctuation across the retinal layers is related to both the
differing capillary densities and disparity in oxygen consumption
by various retinal neurons and glia.
[0944] Local oxygen tension can significantly affect the retina and
its microvasculature by regulation of an array of vasoactive
agents, including, for example, vascular endothelial growth factor
(VEGF). (See, e.g., Werdich et al., Exp. Eye Res. 79:623 (2004);
Arden et al., Br. J. Ophthalmol. 89:764 (2005)). Rod photoreceptors
are believed to have the highest metabolic rate of any cell in the
body (see, e.g., Arden et al., supra). During dark adaptation, the
rod photoreceptors recover their high cytoplasmic calcium levels
via cGMP-gated calcium channels with concomitant extrusion of
sodium ions and water. The efflux of sodium from the cell is an
ATP-dependent process, such that the retinal neurons consume up to
an estimated five times more oxygen under scotopic (i.e., dark
adapted), compared with photopic (i.e., light adapted) conditions.
Thus, during characteristic dark adaptation of photoreceptors, the
high metabolic demand leads to significant local reduction of
oxygen levels in the dark-adapted retina (Ahmed et al, Invest.
Ophthalmol. Vis. Sci. 34:516 (1993)).
[0945] Without being bound by any one theory, retinal hypoxia may
be further increased in the retina of subjects who have diseases or
conditions such as, for example, central retinal vein occlusion in
which the retinal vasculature is already compromised. Increasing
hypoxia may increase susceptibility to sight-threatening, retinal
neovascularization. Neovascularization is the formation of new,
functional microvascular networks with red blood cell perfusion,
and is a characteristic of retinal degenerative disorders,
including, but not limited to, diabetic retinopathy, retinopathy of
prematurity, wet AMD and central retinal vein occlusions.
Preventing or inhibiting dark adaptation of rod photoreceptor
cells, thereby decreasing expenditure of energy and consumption of
oxygen (i.e., reducing metabolic demand), may inhibit or slow
retinal degeneration, and/or may promote regeneration of retinal
cells, including rod photoreceptor cells and retinal pigment
epithelial (RPE) cells, and may reduce hypoxia and may inhibit
neovascularization.
[0946] Methods are described herein for inhibiting (i.e., reducing,
preventing, slowing or retarding, in a biologically or
statistically significant manner) degeneration of retinal cells
(including retinal neuronal cells as described herein and RPE
cells) and/or for reducing (i.e., preventing or slowing,
inhibiting, abrogating in a biologically or statistically
significant manner) retinal ischemia. Methods are also provided for
inhibiting (i.e., reducing, preventing, slowing or retarding, in a
biologically or statistically significant manner)
neovascularization in the eye, particularly in the retina. Such
methods comprise contacting the retina, and thus, contacting
retinal cells (including retinal neuronal cells such as rod
photoreceptor cells, and RPE cells) with at least one of the
compounds described herein that inhibits at least one visual cycle
trans-cis isomerase (which may include inhibition of isomerization
of an all-trans-retinal ester), under conditions and at a time that
may prevent, inhibit, or delay dark adaptation of a rod
photoreceptor cell in the retina. As described in further detail
herein, in particular embodiments, the compound that contacts the
retina interacts with an isomerase enzyme or enzymatic complex in a
RPE cell in the retina and inhibits, blocks, or in some manner
interferes with the catalytic activity of the isomerase. Thus,
isomerization of an all-trans-retinyl ester is inhibited or
reduced. The compounds described herein or compositions comprising
said compounds may be administered to a subject who has developed
and manifested an ophthalmic disease or disorder or who is at risk
of developing an ophthalmic disease or disorder, or to a subject
who presents or who is at risk of presenting a condition such as
retinal neovascularization or retinal ischemia.
[0947] By way of background, the visual cycle (also called retinoid
cycle) refers to the series of enzyme and light-mediated
conversions between the 11-cis and all-trans forms of
retinol/retinal that occur in the photoreceptor and retinal pigment
epithelial (RPE) cells of the eye. In vertebrate photoreceptor
cells, a photon causes isomerization of the 11-cis-retinylidene
chromophore to all-trans-retinylidene coupled to the visual opsin
receptors. This photoisomerization triggers conformational changes
of opsins, which, in turn, initiate the biochemical chain of
reactions termed phototransduction (Filipek et al., Annu. Rev.
Physiol. 65 851-79 (2003)). After absorption of light and
photoisomerization of 11-cis-retinal to all-trans retinal,
regeneration of the visual chromophore is a critical step in
restoring photoreceptors to their dark-adapted state. Regeneration
of the visual pigment requires that the chromophore be converted
back to the 11-cis-configuration (reviewed in McBee et al., Prog.
Retin. Eye Res. 20:469-52 (2001)). The chromophore is released from
the opsin and reduced in the photoreceptor by retinol
dehydrogenases. The product, all-trans-retinol, is trapped in the
adjacent retinal pigment epithelium (RPE) in the form of insoluble
fatty acid esters in subcellular structures known as retinosomes
(Imanishi et al., J. Cell Biol. 164:373-78 (2004)).
[0948] During the visual cycle in rod receptor cells, the 11-cis
retinal chromophore within the visual pigment molecule, which is
called rhodopsin, absorbs a photon of light and is isomerized to
the all-trans configuration, thereby activating the
phototransduction cascade. Rhodopsin is a G-protein coupled
receptor (GPCR) that consists of seven membrane-spanning helices
that are interconnected by extracellular and cytoplasmic loops.
When the all-trans form of the retinoid is still covalently bound
to the pigment molecule, the pigment is referred to as
metarhodopsin, which exists in different forms (e.g., metarhodopsin
I and metarhodopsin II). The all-trans-retinoid is then hydrolyzed
and the visual pigment is in the form of the apoprotein, opsin,
which is also called apo-rhodopsin in the art and herein. This
all-trans-retinoid is transported or chaperoned out of the
photoreceptor cell and across the extracellular space to the RPE
cells, where the retinoid is converted to the 11-cis-isomer. The
movement of the retinoids between the RPE and photoreceptors cells
is believed to be accomplished by different chaperone polypeptides
in each of the cell types. See Lamb et al., Progress in Retinal and
Eye Research 23:307-80 (2004).
[0949] Under light conditions, rhodopsin continually transitions
through the three forms, rhodopsin, metarhodopsin, and
apo-rhodopsin. When most of the visual pigment is in the rhodopsin
form (i.e., bound with 11-cis retinal), the rod photoreceptor cell
is in a "dark-adapted" state. When the visual pigment is
predominantly in the metarhodopsin form (i.e., bound with
all-trans-retinal), the state of the photoreceptor cell is referred
to as a "light-adapted," and when the visual pigment is
apo-rhodopsin (or opsin) and no longer has bound chromophore, the
state of the photoreceptor cell is referred to as
"rhodopsin-depleted." Each of the three states of the photoreceptor
cell has different energy requirements, and differing levels of ATP
and oxygen are consumed. In the dark-adapted state, rhodopsin has
no regulatory effect on cation channels, which are open, resulting
in an influx of cations (Na.sup.+/K.sup.+ and Ca.sup.2+). To
maintain the proper level of these cations in the cell during the
dark state, the photoreceptor cells actively transport the cations
out of the cell via ATP-dependent pumps. Thus maintenance of this
"dark current" requires a large amount of energy, resulting in high
metabolic demand. In the light-adapted state, metarhodopsin
triggers an enzymatic cascade process that results in hydrolysis of
GMP, which in turn, closes cation-specific channels in the
photoreceptor cell membrane. In the rhodopsin-depleted state, the
chromophore is hydrolyzed from metarhodopsin to form the
apoprotein, opsin (apo-rhodopsin), which partially regulates the
cation channels such that the rod photoreceptor cells exhibit an
attenuated current compared with the photoreceptor in the
dark-adapted state, resulting in a moderate metabolic demand.
[0950] Under normal light conditions, the incidence of rod
photoreceptors in the dark adapted state is small, in general, 2%
or less, and the cells are primarily in the light-adapted or
rhodopsin-depleted states, which overall results in a relatively
low metabolic demand compared with cells in the dark-adapted state.
At night, however, the relative incidence of the dark-adapted
photoreceptor state increases profoundly, due to the absence of
light adaptation and to the continued operation of the "dark"
visual cycle in RPE cells, which replenishes the rod photoreceptor
cells with 11-cis-retinal. This shift to dark adaptation of the rod
photoreceptor causes an increase in metabolic demand (that is,
increased ATP and oxygen consumption), leading ultimately to
retinal hypoxia and subsequent initiation of angiogenesis. Most
ischaemic insults to the retina therefore occur in the dark, for
example, at night during sleep.
[0951] Without being bound by any theory, therapeutic intervention
during the "dark" visual cycle may prevent retinal hypoxia and
neovascularization that are caused by high metabolic activity in
the dark-adapted rod photoreceptor cell. Merely by way of one
example, altering the "dark" visual cycle by administering any one
of the compounds described herein, which is an isomerase inhibitor,
rhodopsin (i.e., 11-cis-retinal bound) may be reduced or depleted,
preventing or inhibiting dark adaptation of rod photoreceptors.
This in turn may reduce retinal metabolic demand, attenuating the
nighttime risk of retinal ischemia and neovascularization, and
thereby inhibiting or slowing retinal degeneration.
[0952] In one embodiment, at least one of the compounds described
herein (i.e., a compound as described in detail herein, including a
compound having the structure as set forth in Formula (I), (II),
(IIa), (III), (IIIa), (IV), or (IVa) and substructures thereof, and
the specific compounds described herein) that, for example, blocks,
reduces, inhibits, or in some manner attenuates the catalytic
activity of a visual cycle isomerase in a statistically or
biologically significant manner, may prevent, inhibit, or delay
dark adaptation of a rod photoreceptor cell, thereby inhibiting
(i.e., reducing, abrogating, preventing, slowing the progression
of, or decreasing in a statistically or biologically significant
manner) degeneration of retinal cells (or enhancing survival of
retinal cells) of the retina of an eye. In another embodiment, the
compounds described herein may prevent or inhibit dark adaptation
of a rod photoreceptor cell, thereby reducing ischemia (i.e.,
decreasing, preventing, inhibiting, slowing the progression of
ischemia in a statistically or biologically significant manner). In
yet another embodiment, any one of the compounds described herein
may prevent dark adaptation of a rod photoreceptor cell, thereby
inhibiting neovascularization in the retina of an eye. Accordingly,
methods are provided herein for inhibiting retinal cell
degeneration, for inhibiting neovascularization in the retina of an
eye of a subject, and for reducing ischemia in an eye of a subject
wherein the methods comprise administering at least one compound
described herein, under conditions and at a time sufficient to
prevent, inhibit, or delay dark adaptation of a rod photoreceptor
cell. These methods and compositions are therefore useful for
treating an ophthalmic disease or disorder including, but not
limited to, diabetic retinopathy, diabetic maculopathy, retinal
blood vessel occlusion, retinopathy of prematurity, or ischemia
reperfusion related retinal injury.
[0953] The compounds described herein (i.e., a compound as
described in detail herein, including a compound having the
structure as set forth in Formula (I), (II), (IIa), (III), (IIIa),
(IV), or (IVa), and substructures thereof, and the specific
compounds described herein) may prevent (i.e., delay, slow,
inhibit, or decrease) recovery of the visual pigment chromophore,
which may prevent or inhibit or retard the formation of retinals
and may increase the level of retinal esters, which perturbs the
visual cycle, inhibiting regeneration of rhodopsin, and which
prevents, slows, delays or inhibits dark adaptation of a rod
photoreceptor cell. In certain embodiments, when dark adaptation of
rod photoreceptor cells is prevented in the presence of the
compound, dark adaptation is substantially prevented, and the
number or percent of rod photoreceptor cells that are
rhodopsin-depleted or light adapted is increased compared with the
number or percent of cells that are rhodopsin-depleted or
light-adapted in the absence of the agent. Thus, in certain
embodiments when dark adaptation of rod photoreceptor cells is
prevented (i.e., substantially prevented), only at least 2% of rod
photoreceptor cells are dark-adapted, similar to the percent or
number of cells that are in a dark-adapted state during normal,
light conditions. In other embodiments, at least 5-10%, 10-20%,
20-30%, 30-40%, 40-50%, 50-60%, or 60-70% of rod photoreceptor
cells are dark-adapted after administration of an agent. In other
embodiments, the compound acts to delay dark adaptation, and in the
presence of the compound dark adaptation of rod photoreceptor cells
may be delayed 30 minutes, one hour, two hours, three hours, or
four hours compared to dark adaptation of rod photoreceptors in the
absence of the compound. By contrast, when a compound described
herein is administered such that the compound effectively inhibits
isomerization of substrate during light-adapted conditions, the
compound is administered in such a manner to minimize the percent
of rod photoreceptor cells that are dark-adapted, for example, only
2%, 5%, 10%, 20%, or 25% of rod photoreceptors are dark-adapted
(see e.g., U.S. Patent Application Publication No. 2006/0069078;
Patent Application No. PCT/US2007/002330).
[0954] In the retina in the presence of at least one compound
described herein, regeneration of rhodopsin in a rod photoreceptor
cell may be inhibited or the rate of regeneration may be reduced
(i.e., inhibited, reduced, or decreased in a statistically or
biologically significant manner), at least in part, by preventing
the formation of retinals, reducing the level of retinals, and/or
increasing the level of retinyl esters. To determine the level of
regeneration of rhodopsin in a rod photoreceptor cell, the level of
regeneration of rhodopsin (which may be called a first level) may
be determined prior to permitting contact between the compound and
the retina (i.e., prior to administration of the agent). After a
time sufficient for the compound and the retina and cells of the
retina to interact, (i.e., after administration of the compound),
the level of regeneration of rhodopsin (which may be called a
second level) may be determined. A decrease in the second level
compared with the first level indicates that the compound inhibits
regeneration of rhodopsin. The level of rhodopsin generation may be
determined after each dose, or after any number of doses, and
ongoing throughout the therapeutic regimen to characterize the
effect of the agent on regeneration of rhodopsin.
[0955] In certain embodiments, the subject in need of the
treatments described herein, may have a disease or disorder that
results in or causes impairment of the capability of rod
photoreceptors to regenerate rhodopsin in the retina. By way of
example, inhibition of rhodopsin regeneration (or reduction of the
rate of rhodopsin regeneration) may be symptomatic in patients with
diabetes. In addition to determining the level of regeneration of
rhodopsin in the subject who has diabetes before and after
administration of a compound described herein, the effect of the
compound may also be characterized by comparing inhibition of
rhodopsin regeneration in a first subject (or a first group or
plurality of subjects) to whom the compound is administered, to a
second subject (or second group or plurality of subjects) who has
diabetes but who does not receive the agent.
[0956] In another embodiment, a method is provided for preventing
or inhibiting dark adaptation of a rod photoreceptor cell (or a
plurality of rod photoreceptor cells) in a retina comprising
contacting the retina and at least one of the compounds described
herein (i.e., a compound as described in detail herein, including a
compound having the structure as set forth in Formula (I), (II),
(IIa), (III), (IIIa), (IV), or (IVa), and substructures thereof,
and the specific compounds described herein), under conditions and
at a time sufficient to permit interaction between the agent and an
isomerase present in a retinal cell (such as an RPE cell). A first
level of 11-cis-retinal in a rod photoreceptor cell in the presence
of the compound may be determined and compared to a second level of
11-cis-retinal in a rod photoreceptor cell in the absence of the
compound. Prevention or inhibition of dark adaptation of the rod
photoreceptor cell is indicated when the first level of
11-cis-retinal is less than the second level of 11-cis-retinal.
[0957] Inhibiting regeneration of rhodopsin may also include
increasing the level of 11-cis-retinyl esters present in the RPE
cell in the presence of the compound compared with the level of
11-cis-retinyl esters present in the RPE cell in the absence of the
compound (i.e., prior to administration of the agent). A two-photon
imaging technique may be used to view and analyze retinosome
structures in the RPE, which structures are believed to store
retinyl esters (see, e.g., Imanishi et al., J. Cell Biol.
164:373-83 (2004), Epub 2004 Jan. 26.). A first level of retinyl
esters may be determined prior to administration of the compound,
and a second level of retinyl esters may be determined after
administration of a first dose or any subsequent dose, wherein an
increase in the second level compared to the first level indicates
that the compound inhibits regeneration of rhodopsin.
[0958] Retinyl esters may be analyzed by gradient HPLC according to
methods practiced in the art (see, for example, Mata et al., Neuron
36:69-80 (2002); Trevino et al. J. Exp. Biol. 208:4151-57 (2005)).
To measure 11-cis and all-trans retinals, retinoids may be
extracted by a formaldehyde method (see, e.g., Suzuki et al., Vis.
Res. 28:1061-70 (1988); Okajima and Pepperberg, Exp. Eye Res.
65:331-40 (1997)) or by a hydroxylamine method (see, e.g.,
Groenendijk et al., Biochim. Biophys. Acta. 617:430-38 (1980))
before being analyzed on isocratic HPLC (see, e.g., Trevino et al.,
supra). The retinoids may be monitored spectrophotometrically (see,
e.g., Maeda et al., J. Neurochem. 85:944-956 (2003); Van Hooser et
al., J. Biol. Chem. 277:19173-82 (2002)).
[0959] In another embodiment of the methods described herein for
treating an ophthalmic disease or disorder, for inhibiting retinal
cell degeneration (or enhancing retinal cell survival), for
inhibiting neovascularization, and for reducing ischemia in the
retina, preventing or inhibiting dark adaptation of a rod
photoreceptor cell in the retina comprises increasing the level of
apo-rhodopsin (also called opsin) in the photoreceptor cell. The
total level of the visual pigment approximates the sum of rhodopsin
and apo-rhodopsin and the total level remains constant. Therefore,
preventing, delaying, or inhibiting dark adaptation of the rod
photoreceptor cell may alter the ratio of apo-rhodopsin to
rhodopsin. In particular embodiments, preventing, delaying, or
inhibiting dark adaptation by administering a compound described
herein may increase the ratio of the level of apo-rhodopsin to the
level of rhodopsin compared to the ratio in the absence of the
agent (for example, prior to administration of the agent). An
increase in the ratio (i.e., a statistically or biologically
significant increase) of apo-rhodopsin to rhodopsin indicates that
the percent or number of rod photoreceptor cells that are
rhodopsin-depleted is increased and that the percent or number of
rod photoreceptor cells that are dark-adapted is decreased. The
ratio of apo-rhodopsin to rhodopsin may be determined throughout
the course of therapy to monitor the effect of the agent.
[0960] Determining or characterizing the capability of compound to
prevent, delay, or inhibit dark adaptation of a rod photoreceptor
cell may be determined in animal model studies. The level of
rhodopsin and the ratio of apo-rhodopsin to rhodopsin may be
determined prior to administration (which may be called a first
level or first ratio, respectively) of the agent and then after
administration of a first or any subsequent dose of the agent
(which may be called a second level or second ratio, respectively)
to determine and to demonstrate that the level of apo-rhodopsin is
greater than the level of apo-rhodopsin in the retina of animals
that did not receive the agent. The level of rhodopsin in rod
photoreceptor cells may be performed according to methods practiced
in the art and provided herein (see, e.g., Yan et al. J. Biol.
Chem. 279:48189-96 (2004)).
[0961] A subject in need of such treatment may be a human or may be
a non-human primate or other animal (i.e., veterinary use) who has
developed symptoms of an ophthalmic disease or disorder or who is
at risk for developing an ophthalmic disease or disorder. Examples
of non-human primates and other animals include but are not limited
to farm animals, pets, and zoo animals (e.g., horses, cows,
buffalo, llamas, goats, rabbits, cats, dogs, chimpanzees,
orangutans, gorillas, monkeys, elephants, bears, large cats,
etc.).
[0962] Also provided herein are methods for inhibiting (reducing,
slowing, preventing) degeneration and enhancing retinal neuronal
cell survival (or prolonging cell viability) comprising
administering to a subject a composition comprising a
pharmaceutically acceptable carrier and a compound described in
detail herein, including a compound having any one of the
structures set forth in Formula (I), (II), (IIa), (III), (IIIa),
(IV), or (IVa) and substructures thereof, and specific compounds
recited herein. Retinal neuronal cells include photoreceptor cells,
bipolar cells, horizontal cells, ganglion cells, and amacrine
cells. In another embodiment, methods are provided for enhancing
survival or inhibiting degeneration of a mature retinal cell such
as a RPE cell or a Muller glial cell. In other embodiments, a
method for preventing or inhibiting photoreceptor degeneration in
an eye of a subject are provided. A method that prevents or
inhibits photoreceptor degeneration may include a method for
restoring photoreceptor function in an eye of a subject. Such
methods comprise administering to the subject a composition
comprising a compound as described herein and a pharmaceutically or
acceptable carrier (i.e., excipient or vehicle). More specifically,
these methods comprise administering to a subject a
pharmaceutically acceptable excipient and a compound described
herein, including a compound having any one of the structures set
forth in Formula (I), (II), (IIa), (III), (IIIa), (IV), or (IVa) or
substructures thereof described herein. Without wishing to be bound
by theory, the compounds described herein may inhibit an
isomerization step of the retinoid cycle (i.e., visual cycle)
and/or may slow chromophore flux in a retinoid cycle in the
eye.
[0963] The ophthalmic disease may result, at least in part, from
lipofuscin pigment(s) accumulation and/or from accumulation of
N-retinylidene-N-retinylethanolamine (A2E) in the eye. Accordingly,
in certain embodiments, methods are provided for inhibiting or
preventing accumulation of lipofuscin pigment(s) and/or A2E in the
eye of a subject. These methods comprise administering to the
subject a composition comprising a pharmaceutically acceptable
carrier and a compound as described in detail herein, including a
compound having the structure as set forth in Formula (I), (II),
(IIa), (III), (IIIa), (IV), or (IVa) or substructures thereof.
[0964] A compound described herein can be administered to a subject
who has an excess of a retinoid in an eye (e.g., an excess of
11-cis-retinol or 11-cis-retinal), an excess of retinoid waste
products or intermediates in the recycling of all-trans-retinal, or
the like. Methods described herein and practiced in the art may be
used to determine whether the level of one or more endogenous
retinoids in a subject are altered (increased or decreased in a
statistically significant or biologically significant manner)
during or after administration of any one of the compounds
described herein. Rhodopsin, which is composed of the protein opsin
and retinal (a vitamin A form), is located in the membrane of the
photoreceptor cell in the retina of the eye and catalyzes the only
light-sensitive step in vision. The 11-cis-retinal chromophore lies
in a pocket of the protein and is isomerized to all-trans-retinal
when light is absorbed. The isomerization of retinal leads to a
change of the shape of rhodopsin, which triggers a cascade of
reactions that lead to a nerve impulse that is transmitted to the
brain by the optic nerve.
[0965] Methods of determining endogenous retinoid levels in a
vertebrate eye, and an excess or deficiency of such retinoids, are
disclosed in, for example, U.S. Patent Application Publication No:
2005/0159662 (the disclosure of which is incorporated by reference
herein in its entirety). Other methods of determining endogenous
retinoid levels in a subject, which is useful for determining
whether levels of such retinoids are above the normal range, and
include for example, analysis by high pressure liquid
chromatography (HPLC) of retinoids in a biological sample from a
subject. For example, retinoid levels can be determined in a
biological sample that is a blood sample (which includes serum or
plasma) from a subject. A biological sample may also include
vitreous fluid, aqueous humor, intraocular fluid, subretinal fluid,
or tears.
[0966] For example, a blood sample can be obtained from a subject,
and different retinoid compounds and levels of one or more of the
retinoid compounds in the sample can be separated and analyzed by
normal phase high pressure liquid chromatography (HPLC) (e.g., with
a HP1100 HPLC and a Beckman, Ultrasphere-Si, 4.6 mm.times.250 mm
column using 10% ethyl acetate/90% hexane at a flow rate of 1.4
ml/minute). The retinoids can be detected by, for example,
detection at 325 nm using a diode-array detector and HP Chemstation
A.03.03 software. An excess in retinoids can be determined, for
example, by comparison of the profile of retinoids (i.e.,
qualitative, e.g., identity of specific compounds, and
quantitative, e.g., the level of each specific compound) in the
sample with a sample from a normal subject. Persons skilled in the
art who are familiar with such assays and techniques and will
readily understand that appropriate controls are included.
[0967] As used herein, increased or excessive levels of endogenous
retinoid, such as 11-cis-retinol or 11-cis-retinal, refer to levels
of endogenous retinoid higher than those found in a healthy eye of
a young vertebrate of the same species. Administration of a
compound described herein can reduce or eliminate the requirement
for endogenous retinoid. In certain embodiments, the level of
endogenous retinoid may be compared before and after any one or
more doses of a compound described herein is administered to a
subject to determine the effect of the compound on the level of
endogenous retinoids in the subject.
[0968] In another embodiment, the methods described herein for
treating an ophthalmic disease or disorder, for inhibiting
neovascularization, and for reducing ischemia in the retina
comprise administering at least one of the compounds described
herein, thereby effecting a decrease in metabolic demand, which
includes effecting a reduction in ATP consumption and in oxygen
consumption in rod photoreceptor cells. As described herein,
consumption of ATP and oxygen in a dark-adapted rod photoreceptor
cell is greater than in rod photoreceptor cells that are
light-adapted or rhodopsin-depleted; thus, use of the compounds in
the methods described herein may reduce the consumption of ATP in
the rod photoreceptor cells that are prevented, inhibited, or
delayed from dark adaptation compared with rod photoreceptor cells
that are dark-adapted (such as the cells prior to administration or
contact with the compound or cells that are never exposed to the
compound).
[0969] The methods described herein that may prevent or inhibit
dark adaptation of a rod photoreceptor cell may therefore reduce
hypoxia (i.e., reduce in a statistically or biologically
significant manner) in the retina. For example, the level of
hypoxia (a first level) may be determined prior to initiation of
the treatment regimen, that is, prior to the first dosing of the
compound (or a composition, as described herein, comprising the
compound). The level of hypoxia (for example, a second level) may
be determined after the first dosing, and/or after any second or
subsequent dosing to monitor and characterize hypoxia throughout
the treatment regimen. A decrease (reduction) in the second (or any
subsequent) level of hypoxia compared to the level of hypoxia prior
to initial administration indicates that the compound and the
treatment regiment prevent dark adaptation of the rod photoreceptor
cells and may be used for treating ophthalmic diseases and
disorders. Consumption of oxygen, oxygenation of the retina, and/or
hypoxia in the retina may be determined using methods practiced in
the art. For example, oxygenation of the retina may be determined
by measuring the fluorescence of flavoproteins in the retina (see,
e.g., U.S. Pat. No. 4,569,354). Another exemplary method is retinal
oximetry that measures blood oxygen saturation in the large vessels
of the retina near the optic disc. Such methods may be used to
identify and determine the extent of retinal hypoxia before changes
in retinal vessel architecture can be detected.
[0970] A biological sample may be a blood sample (from which serum
or plasma may be prepared), biopsy specimen, body fluids (e.g.,
vitreous fluid, aqueous humor, intraocular fluid, subretinal fluid,
or tears), tissue explant, organ culture, or any other tissue or
cell preparation from a subject or a biological source. A sample
may further refer to a tissue or cell preparation in which the
morphological integrity or physical state has been disrupted, for
example, by dissection, dissociation, solubilization,
fractionation, homogenization, biochemical or chemical extraction,
pulverization, lyophilization, sonication, or any other means for
processing a sample derived from a subject or biological source.
The subject or biological source may be a human or non-human
animal, a primary cell culture (e.g., a retinal cell culture), or
culture adapted cell line, including but not limited to,
genetically engineered cell lines that may contain chromosomally
integrated or episomal recombinant nucleic acid sequences,
immortalized or immortalizable cell lines, somatic cell hybrid cell
lines, differentiated or differentiatable cell lines, transformed
cell lines, and the like. Mature retinal cells, including retinal
neuronal cells, RPE cells, and Muller glial cells, may be present
in or isolated from a biological sample as described herein. For
example, the mature retinal cell may be obtained from a primary or
long-term cell culture or may be present in or isolated from a
biological sample obtained from a subject (human or non-human
animal).
[0971] 3. Retinal Cells
[0972] The retina is a thin layer of nervous tissue located between
the vitreous body and choroid in the eye. Major landmarks in the
retina are the fovea, the macula, and the optic disc. The retina is
thickest near the posterior sections and becomes thinner near the
periphery. The macula is located in the posterior retina and
contains the fovea and foveola. The foveola contains the area of
maximal cone density and, thus, imparts the highest visual acuity
in the retina. The foveola is contained within the fovea, which is
contained within the macula.
[0973] The peripheral portion of the retina increases the field of
vision. The peripheral retina extends anterior to the ciliary body
and is divided into four regions: the near periphery (most
posterior), the mid-periphery, the far periphery, and the ora
serrata (most anterior). The ora serrata denotes the termination of
the retina.
[0974] The term neuron (or nerve cell) as understood in the art and
used herein denotes a cell that arises from neuroepithelial cell
precursors. Mature neurons (i.e. fully differentiated cells)
display several specific antigenic markers. Neurons may be
classified functionally into three groups: (1) afferent neurons (or
sensory neurons) that transmit information into the brain for
conscious perception and motor coordination; (2) motor neurons that
transmit commands to muscles and glands; and (3) interneurons that
are responsible for local circuitry; and (4) projection
interneurons that relay information from one region of the brain to
another region and therefore have long axons. Interneurons process
information within specific subregions of the brain and have
relatively shorter axons. A neuron typically has four defined
regions: the cell body (or soma); an axon; dendrites; and
presynaptic terminals. The dendrites serve as the primary input of
information from other neural cells. The axon carries the
electrical signals that are initiated in the cell body to other
neurons or to effector organs. At the presynaptic terminals, the
neuron transmits information to another cell (the postsynaptic
cell), which may be another neuron, a muscle cell, or a secretory
cell.
[0975] The retina is composed of several types of neuronal cells.
As described herein, the types of retinal neuronal cells that may
be cultured in vitro by this method include photoreceptor cells,
ganglion cells, and interneurons such as bipolar cells, horizontal
cells, and amacrine cells. Photoreceptors are specialized
light-reactive neural cells and comprise two major classes, rods
and cones. Rods are involved in scotopic or dim light vision,
whereas photopic or bright light vision originates in the cones.
Many neurodegenerative diseases, such as AMD, that result in
blindness affect photoreceptors.
[0976] Extending from their cell bodies, the photoreceptors have
two morphologically distinct regions, the inner and outer segments.
The outer segment lies furthermost from the photoreceptor cell body
and contains disks that convert incoming light energy into
electrical impulses (phototransduction). The outer segment is
attached to the inner segment with a very small and fragile cilium.
The size and shape of the outer segments vary between rods and
cones and are dependent upon position within the retina. See Hogan,
"Retina" in Histology of the Human Eye: an Atlas and Text Book
(Hogan et al. (eds). WB Saunders; Philadelphia, Pa. (1971)); Eye
and Orbit, 8.sup.th Ed., Bron et al., (Chapman and Hall, 1997).
[0977] Ganglion cells are output neurons that convey information
from the retinal interneurons (including horizontal cells, bipolar
cells, amacrine cells) to the brain. Bipolar cells are named
according to their morphology, and receive input from the
photoreceptors, connect with amacrine cells, and send output
radially to the ganglion cells. Amacrine cells have processes
parallel to the plane of the retina and have typically inhibitory
output to ganglion cells. Amacrine cells are often subclassified by
neurotransmitter or neuromodulator or peptide (such as calretinin
or calbindin) and interact with each other, with bipolar cells, and
with photoreceptors. Bipolar cells are retinal interneurons that
are named according to their morphology; bipolar cells receive
input from the photoreceptors and sent the input to the ganglion
cells. Horizontal cells modulate and transform visual information
from large numbers of photoreceptors and have horizontal
integration (whereas bipolar cells relay information radially
through the retina).
[0978] Other retinal cells that may be present in the retinal cell
cultures described herein include glial cells, such as Muller glial
cells, and retinal pigment epithelial cells (RPE). Glial cells
surround nerve cell bodies and axons. The glial cells do not carry
electrical impulses but contribute to maintenance of normal brain
function. Muller glia, the predominant type of glial cell within
the retina, provide structural support of the retina and are
involved in the metabolism of the retina (e.g., contribute to
regulation of ionic concentrations, degradation of
neurotransmitters, and remove certain metabolites (see, e.g.,
Kljavin et al., J. Neurosci. 11:2985 (1991))). Muller's fibers
(also known as sustentacular fibers of retina) are sustentacular
neuroglial cells of the retina that run through the thickness of
the retina from the internal limiting membrane to the bases of the
rods and cones where they form a row of junctional complexes.
[0979] Retinal pigment epithelial (RPE) cells form the outermost
layer of the retina, separated from the blood vessel-enriched
choroids by Bruch's membrane. RPE cells are a type of phagocytic
epithelial cell, with some functions that are macrophage-like,
which lies immediately below the retinal photoreceptors. The dorsal
surface of the RPE cell is closely apposed to the ends of the rods,
and as discs are shed from the rod outer segment they are
internalized and digested by RPE cells. Similar process occurs with
the disc of the cones. RPE cells also produce, store, and transport
a variety of factors that contribute to the normal function and
survival of photoreceptors. Another function of RPE cells is to
recycle vitamin A as it moves between photoreceptors and the RPE
during light and dark adaptation in the process known as the visual
cycle.
[0980] Described herein is an exemplary long-term in vitro cell
culture system permits and promotes the survival in culture of
mature retinal cells, including retinal neurons, for at least 2-4
weeks, over 2 months, or for as long as 6 months. The cell culture
system may be used for identifying and characterizing the compounds
described herein that are useful in the methods described herein
for treating and/or preventing an ophthalmic disease or disorder or
for preventing or inhibiting accumulation in the eye of
lipofuscin(s) and/or A2E. Retinal cells are isolated from
non-embryonic, non-tumorigenic tissue and have not been
immortalized by any method such as, for example, transformation or
infection with an oncogenic virus. The cell culture system
comprises all the major retinal neuronal cell types
(photoreceptors, bipolar cells, horizontal cells, amacrine cells,
and ganglion cells), and also may include other mature retinal
cells such as retinal pigment epithelial cells and Muller glial
cells.
[0981] For example, a blood sample can be obtained from a subject,
and different retinoid compounds and levels of one or more of the
retinoid compounds in the sample can be separated and analyzed by
normal phase high pressure liquid chromatography (HPLC) (e.g., with
a HP1100 HPLC and a Beckman, Ultrasphere-Si, 4.6 mm.times.250 mm
column using 10% ethyl acetate/90% hexane at a flow rate of 1.4
ml/minute). The retinoids can be detected by, for example,
detection at 325 nm using a diode-array detector and HP Chemstation
A.03.03 software. An excess in retinoids can be determined, for
example, by comparison of the profile of retinoids (i.e.,
qualitative, e.g., identity of specific compounds, and
quantitative, e.g., the level of each specific compound) in the
sample with a sample from a normal subject. Persons skilled in the
art who are familiar with such assays and techniques and will
readily understand that appropriate controls are included.
[0982] As used herein, increased or excessive levels of endogenous
retinoid, such as 11-cis-retinol or 11-cis-retinal, refer to levels
of endogenous retinoid higher than those found in a healthy eye of
a young vertebrate of the same species. Administration of a
compound described herein can reduce or eliminate the requirement
for endogenous retinoid.
[0983] 4. In Vivo and In Vitro Methods for Determining Therapeutic
Effectiveness of Compounds
[0984] In one embodiment, methods are provided for using the
compounds described herein for enhancing or prolonging retinal cell
survival, including retinal neuronal cell survival and RPE cell
survival. Also provided herein are methods for inhibiting or
preventing degeneration of a retinal cell, including a retinal
neuronal cell (e.g., a photoreceptor cell, an amacrine cell, a
horizontal cell, a bipolar cell, and a ganglion cell) and other
mature retinal cells such as retinal pigment epithelial cells and
Muller glial cells using the compounds described herein. Such
methods comprise, in certain embodiments, administration of a
compound as described herein. Such a compound is useful for
enhancing retinal cell survival, including photoreceptor cell
survival and retinal pigment epithelia survival, inhibiting or
slowing degeneration of a retinal cell, and thus increasing retinal
cell viability, which can result in slowing or halting the
progression of an ophthalmic disease or disorder or retinal injury,
which are described herein.
[0985] The effect of a compound described herein on retinal cell
survival (and/or retinal cell degeneration) may be determined by
using cell culture models, animal models, and other methods that
are described herein and practiced by persons skilled in the art.
By way of example, and not limitation, such methods and assays
include those described in Oglivie et al., Exp. Neurol. 161:675-856
(2000); U.S. Pat. No. 6,406,840; WO 01/81551; WO 98/12303; U.S.
Patent Application No. 2002/0009713; WO 00/40699; U.S. Pat. No.
6,117,675; U.S. Pat. No. 5,736,516; WO 99/29279; WO 01/83714; WO
01/42784; U.S. Pat. No. 6,183,735; U.S. Pat. No. 6,090,624; WO
01/09327; U.S. Pat. No. 5,641,750; U.S. Patent Application
Publication No. 2004/0147019; and U.S. Patent Application
Publication No. 2005/0059148.
[0986] Compounds described herein that may be useful for treating
an ophthalmic disease or disorder (including a retinal disease or
disorder) may inhibit, block, impair, or in some manner interfere
with one or more steps in the visual cycle (also called the
retinoid cycle herein and in the art). Without wishing to be bound
by a particular theory, a compound described herein may inhibit or
block an isomerization step in the visual cycle, for example, by
inhibiting or blocking a functional activity of a visual cycle
trans-cis isomerase. The compounds described herein may inhibit,
directly or indirectly, isomerization of all-trans-retinol to
11-cis-retinol. The compounds may bind to, or in some manner
interact with, and inhibit the isomerase activity of at least one
isomerase in a retinal cell. Any one of the compounds described
herein may also directly or indirectly inhibit or reduce the
activity of an isomerase that is involved in the visual cycle. The
compound may block or inhibit the capability of the isomerase to
bind to one or more substrates, including but not limited to, an
all-trans-retinal ester substrate or all-trans-retinol.
Alternatively, or in addition, the compound may bind to the
catalytic site or region of the isomerase, thereby inhibiting the
capability of the enzyme to catalyze isomerization of at least one
substrate. On the basis of scientific data to date, an at least one
isomerase that catalyzes the isomerization of a substrate during
the visual cycle is believed to be located in the cytoplasm of RPE
cells. As discussed herein, each step, enzyme, substrate,
intermediate, and product of the visual cycle is not yet
elucidated. While a polypeptide called RPE65, which has been found
in the cytoplasm and membrane bound in RPE cells, is hypothesized
to have isomerase activity (and has also been referred to in the
art as having isomerohydrolase activity) (see, e.g., Moiseyev et
al., Proc. Natl. Acad. Sci. USA 102:12413-18 (2004); Chen et al.,
Invest. Ophthalmol. Vis. Sci. 47:1177-84 (2006)), other persons
skilled in the art believe that the RPE65 acts primarily as a
chaperone for all-trans-retinal esters (see, e.g., Lamb et al.
supra).
[0987] Exemplary methods are described herein and practiced by
persons skilled in the art for determining the level of enzymatic
activity of a visual cycle isomerase in the presence of any one of
the compounds described herein. A compound that decreases isomerase
activity may be useful for treating an ophthalmic disease or
disorder. Thus, methods are provided herein for detecting
inhibition of isomerase activity comprising contacting (i.e.,
mixing, combining, or in some manner permitting the compound and
isomerase to interact) a biological sample comprising the isomerase
and a compound described herein and then determining the level of
enzymatic activity of the isomerase. A person having skill in the
art will appreciate that as a control, the level of activity of the
isomerase in the absence of a compound or in the presence of a
compound known not to alter the enzymatic activity of the isomerase
can be determined and compared to the level of activity in the
presence of the compound. A decrease in the level of isomerase
activity in the presence of the compound compared to the level of
isomerase activity in the absence of the compound indicates that
the compound may be useful for treating an ophthalmic disease or
disorder, such as age-related macular degeneration or Stargardt's
disease. A decrease in the level of isomerase activity in the
presence of the compound compared to the level of isomerase
activity in the absence of the compound indicates that the compound
may also be useful in the methods described herein for inhibiting
or preventing dark adaptation, inhibiting neovascularization and
reducing hypoxia and thus useful for treating an ophthalmic disease
or disorder, for example, diabetic retinopathy, diabetic
maculopathy, retinal blood vessel occlusion, retinopathy of
prematurity, or ischemia reperfusion related retinal injury.
[0988] The capability of a compound described herein to inhibit or
to prevent dark adaptation of a rod photoreceptor cell by
inhibiting regeneration of rhodopsin may be determined by in vitro
assays and/or in vivo animal models. By way of example, inhibition
of regeneration may be determined in a mouse model in which a
diabetes-like condition is induced chemically or in a diabetic
mouse model (see, e.g., Phipps et al., Invest. Ophthalmol. Vis.
Sci. 47:3187-94 (2006); Ramsey et al., Invest. Ophthalmol. Vis.
Sci. 47:5116-24 (2006)). The level of rhodopsin (a first level) may
be determined (for example, spectrophotometrically) in the retina
of animals prior to administration of the agent and compared with
the level (a second level) of rhodopsin measured in the retina of
animals after administration of the agent. A decrease in the second
level of rhodopsin compared with the first level of rhodopsin
indicates that the agent inhibits regeneration of rhodopsin. The
appropriate controls and study design to determine whether
regeneration of rhodopsin is inhibited in a statistically
significant or biologically significant manner can be readily
determined and implemented by persons skilled in the art.
[0989] Methods and techniques for determining or characterizing the
effect of any one of the compounds described herein on dark
adaptation and rhodopsin regeneration in rod photoreceptor cells in
a mammal, including a human, may be performed according to
procedures described herein and practiced in the art. For example,
detection of a visual stimulus after exposure to light (i.e.,
photobleaching) versus time in darkness may be determined before
administration of the first dose of the compound and at a time
after the first dose and/or any subsequent dose. A second method
for determining prevention or inhibition of dark adaptation by the
rod photoreceptor cells includes measurement of the amplitude of at
least one, at least two, at least three, or more electroretinogram
components, which include, for example, the a-wave and the b-wave.
See, for example, Lamb et al., supra; Asi et al., Documenta
Ophthalmologica 79:125-39 (1992).
[0990] Inhibiting regeneration of rhodopsin by a compound described
herein comprises reducing the level of the chromophore,
11-cis-retinal, that is produced and present in the RPE cell, and
consequently reducing the level of 11-cis-retinal that is present
in the photoreceptor cell. Thus, the compound, when permitted to
contact the retina under suitable conditions and at a time
sufficient to prevent dark adaptation of a rod photoreceptor cell
and to inhibit regeneration of rhodopsin in the rod photoreceptor
cell, effects a reduction in the level of 11-cis-retinal in a rod
photoreceptor cell (i.e., a statistically significant or
biologically significant reduction). That is, the level of
11-cis-retinal in a rod photoreceptor cell is greater prior to
administration of the compound when compared with the level of
11-cis-retinal in the photoreceptor cell after the first and/or any
subsequent administration of the compound. A first level of
11-cis-retinal may be determined prior to administration of the
compound, and a second level of 11-cis-retinal may be determined
after administration of a first dose or any subsequent dose to
monitor the effect of the compound. A decrease in the second level
compared to the first level indicates that the compound inhibits
regeneration of rhodopsin and thus inhibits or prevents dark
adaptation of the rod photoreceptor cells.
[0991] An exemplary method for determining or characterizing the
capability of a compound described herein to reduce retinal hypoxia
includes measuring the level of retinal oxygenation, for example,
by Magnetic Resonance Imaging (MRI) to measure changes in oxygen
pressure (see, e.g., Luan et al., Invest. Ophthalmol. Vis. Sci.
47:320-28 (2006)). Methods are also available and routinely
practiced in the art to determine or characterize the capability of
compounds described herein to inhibit degeneration of a retinal
cell (see, e.g., Wenzel et al., Prog. Retin. Eye Res. 24:275-306
(2005)).
[0992] Animal models may be used to characterize and identify
compounds that may be used to treat retinal diseases and disorders.
A recently developed animal model may be useful for evaluating
treatments for macular degeneration has been described by Ambati et
al. (Nat. Med. 9:1390-97 (2003); Epub 2003 Oct. 19). This animal
model is one of only a few exemplary animal models presently
available for evaluating a compound or any molecule for use in
treating (including preventing) progression or development of a
retinal disease or disorder. Animal models in which the ABCR gene,
which encodes an ATP-binding cassette transporter located in the
rims of photoreceptor outer segment discs, may be used to evaluate
the effect of a compound. Mutations in the ABCR gene are associated
with Stargardt's disease, and heterozygous mutations in ABCR have
been associated with AMD. Accordingly, animals have been generated
with partial or total loss of ABCR function and may used to
characterize the compounds described herein. (See, e.g., Mata et
al., Invest. Ophthalmol. Sci. 42:1685-90 (2001); Weng et al., Cell
98:13-23 (1999); Mata et al., Proc. Natl. Acad. Sci. USA 97:7154-49
(2000); US 2003/0032078; U.S. Pat. No. 6,713,300). Other animal
models include the use of mutant ELOVL4 transgenic mice to
determine lipofuscin accumulation, electrophysiology, and
photoreceptor degeneration, or prevention or inhibition thereof
(see, e.g., Karan et al., Proc. Natl. Acad. Sci. USA 102:4164-69
(2005)).
[0993] The effect of any one of the compounds described herein may
be determined in a diabetic retinopathy animal model, such as
described in Luan et al. or may be determined in a normal animal
model, in which the animals have been light or dark adapted in the
presence and absence of any one of the compounds described herein.
Another exemplary method for determining the capability of the
agent to reduce retinal hypoxia measures retinal hypoxia by
deposition of a hydroxyprobe (see, e.g., de Gooyer et al. (Invest.
Ophthalmol. Vis. Sci. 47:5553-60 (2006)). Such a technique may be
performed in an animal model using Rho.sup.-/Rho.sup.- knockout
mice (see de Gooyer et al., supra) in which at least one compound
described herein is administered to group(s) of animals in the
presence and absence of the at least one compound, or may be
performed in normal, wildtype animals in which at least one
compound described herein is administered to group(s) of animals in
the presence and absence of the at least one compound. Other animal
models include models for determining photoreceptor function, such
as rat models that measure elctroretinographic (ERG) oscillatory
potentials (see, e.g., Liu et al., Invest. Ophthalmol. Vis. Sci.
47:5447-52 (2006); Akula et al., Invest. Ophthalmol. Vis. Sci.
48:4351-59 (2007); Liu et al., Invest. Ophthalmol. Vis. Sci.
47:2639-47 (2006); Dembinska et al., Invest. Ophthalmol. Vis. Sci.
43:2481-90 (2002); Penn et al., Invest. Ophthalmol. Vis. Sci.
35:3429-35 (1994); Hancock et al., Invest. Ophthalmol. Vis. Sci.
45:1002-1008 (2004)).
[0994] A method for determining the effect of a compound on
isomerase activity may be performed in vitro as described herein
and in the art (Stecher et al., J. Biol. Chem. 274:8577-85 (1999);
see also Golczak et al., Proc. Natl. Acad. Sci. USA 102:8162-67
(2005)). Retinal pigment epithelium (RPE) microsome membranes
isolated from an animal (such as bovine, porcine, human, for
example) may serve as the source of the isomerase. The capability
of the compounds described herein to inhibit isomerase may also be
determined by an in vivo murine isomerase assay. Brief exposure of
the eye to intense light ("photobleaching" of the visual pigment or
simply "bleaching") is known to photo-isomerize almost all
11-cis-retinal in the retina. The recovery of 11-cis-retinal after
bleaching can be used to estimate the activity of isomerase in vivo
(see, e.g., Maeda et al., J. Neurochem. 85:944-956 (2003); Van
Hooser et al., J. Biol. Chem. 277:19173-82, 2002).
Electroretinographic (ERG) recording may be performed as previously
described (Haeseleer et al., Nat. Neurosci. 7:1079-87 (2004);
Sugitomo et al., J. Toxicol. Sci. 22 Suppl 2:315-25 (1997); Keating
et al., Documenta Ophthalmologica 100:77-92 (2000)). See also
Deigner et al., Science, 244: 968-971 (1989); Gollapalli et al.,
Biochim. Biophys. Acta 1651: 93-101 (2003); Parish, et al., Proc.
Natl. Acad. Sci. USA 95:14609-13 (1998); Radu et al., Proc Natl
Acad Sci USA 101: 5928-33 (2004).
[0995] Cell culture methods, such as the method described herein,
are also useful for determining the effect of a compound described
herein on retinal neuronal cell survival. Exemplary cell culture
models are described herein and described in detail in U.S. Patent
Application Publication No. US 2005-0059148 and U.S. Patent
Application Publication No. US2004-0147019 (which are incorporated
by reference in their entirety), which are useful for determining
the capability of a compound as described herein to enhance or
prolong survival of neuronal cells, particularly retinal neuronal
cells, and of retinal pigment epithelial cells, and inhibit,
prevent, slow, or retard degeneration of an eye, or the retina or
retinal cells thereof, or the RPE, and which compounds are useful
for treating ophthalmic diseases and disorders.
[0996] The cell culture model comprises a long-term or extended
culture of mature retinal cells, including retinal neuronal cells
(e.g., photoreceptor cells, amacrine cells, ganglion cells,
horizontal cells, and bipolar cells). The cell culture system and
methods for producing the cell culture system provide extended
culture of photoreceptor cells. The cell culture system may also
comprise retinal pigment epithelial (RPE) cells and Muller glial
cells.
[0997] The retinal cell culture system may also comprise a cell
stressor. The application or the presence of the stressor affects
the mature retinal cells, including the retinal neuronal cells, in
vitro, in a manner that is useful for studying disease pathology
that is observed in a retinal disease or disorder. The cell culture
model provides an in vitro neuronal cell culture system that will
be useful in the identification and biological testing of a
compound described herein that is suitable for treatment of
neurological diseases or disorders in general, and for treatment of
degenerative diseases of the eye and brain in particular. The
ability to maintain primary, in vitro-cultured cells from mature
retinal tissue, including retinal neurons over an extended period
of time in the presence of a stressor enables examination of
cell-to-cell interactions, selection and analysis of neuroactive
compounds and materials, use of a controlled cell culture system
for in vitro CNS and ophthalmic tests, and analysis of the effects
on single cells from a consistent retinal cell population.
[0998] The cell culture system and the retinal cell stress model
comprise cultured mature retinal cells, retinal neurons, and a
retinal cell stressor, which may be used for screening and
characterizing a compound described herein that are capable of
inducing or stimulating the regeneration of CNS tissue that has
been damaged by disease. The cell culture system provides a mature
retinal cell culture that is a mixture of mature retinal neuronal
cells and non-neuronal retinal cells. The cell culture system
comprises all the major retinal neuronal cell types
(photoreceptors, bipolar cells, horizontal cells, amacrine cells,
and ganglion cells), and may also include other mature retinal
cells such as RPE and Muller glial cells. By incorporating these
different types of cells into the in vitro culture system, the
system essentially resembles an "artificial organ" that is more
akin to the natural in vivo state of the retina.
[0999] Viability of one or more of the mature retinal cell types
that are isolated (harvested) from retinal tissue and plated for
tissue culture may be maintained for an extended period of time,
for example, from two weeks up to six months. Viability of the
retinal cells may be determined according to methods described
herein and known in the art. Retinal neuronal cells, similar to
neuronal cells in general, are not actively dividing cells in vivo
and thus cell division of retinal neuronal cells would not
necessarily be indicative of viability. An advantage of the cell
culture system is the ability to culture amacrine cells,
photoreceptors, and associated ganglion projection neurons and
other mature retinal cells for extended periods of time, thereby
providing an opportunity to determine the effectiveness of a
compound described herein for treatment of retinal disease.
[1000] The biological source of the retinal cells or retinal tissue
may be mammalian (e.g., human, non-human primate, ungulate, rodent,
canine, porcine, bovine, or other mammalian source), avian, or from
other genera. Retinal cells including retinal neurons from
post-natal non-human primates, post-natal pigs, or post-natal
chickens may be used, but any adult or post-natal retinal tissue
may be suitable for use in this retinal cell culture system.
[1001] In certain instances, the cell culture system may provide
for robust long-term survival of retinal cells without inclusion of
cells derived from or isolated or purified from non-retinal tissue.
Such a cell culture system comprises cells isolated solely from the
retina of the eye and thus is substantially free of types of cells
from other parts or regions of the eye that are separate from the
retina, such as the ciliary body, iris, choroid, and vitreous.
Other cell culture methods include the addition of non-retinal
cells, such as ciliary body cell and/or stem cells (which may or
may not be retinal stem cells) and/or additional purified glial
cells.
[1002] The in vitro retinal cell culture systems described herein
may serve as physiological retinal models that can be used to
characterize aspects of the physiology of the retina. This
physiological retinal model may also be used as a broader general
neurobiology model. A cell stressor may be included in the model
cell culture system. A cell stressor, which as described herein is
a retinal cell stressor, adversely affects the viability or reduces
the viability of one or more of the different retinal cell types,
including types of retinal neuronal cells, in the cell culture
system. A person skilled in the art would readily appreciate and
understand that as described herein a retinal cell that exhibits
reduced viability means that the length of time that a retinal cell
survives in the cell culture system is reduced or decreased
(decreased lifespan) and/or that the retinal cell exhibits a
decrease, inhibition, or adverse effect of a biological or
biochemical function (e.g., decreased or abnormal metabolism;
initiation of apoptosis; etc.) compared with a retinal cell
cultured in an appropriate control cell system (e.g., the cell
culture system described herein in the absence of the cell
stressor). Reduced viability of a retinal cell may be indicated by
cell death; an alteration or change in cell structure or
morphology; induction and/or progression of apoptosis; initiation,
enhancement, and/or acceleration of retinal neuronal cell
neurodegeneration (or neuronal cell injury).
[1003] Methods and techniques for determining cell viability are
described in detail herein and are those with which skilled
artisans are familiar. These methods and techniques for determining
cell viability may be used for monitoring the health and status of
retinal cells in the cell culture system and for determining the
capability of the compounds described herein to alter (preferably
increase, prolong, enhance, improve) retinal cell or retinal
pigment epithelial cell viability or retinal cell survival.
[1004] The addition of a cell stressor to the cell culture system
is useful for determining the capability of a compound described
herein to abrogate, inhibit, eliminate, or lessen the effect of the
stressor. The retinal cell culture system may include a cell
stressor that is chemical (e.g., A2E, cigarette smoke concentrate);
biological (for example, toxin exposure; beta-amyloid;
lipopolysaccharides); or non-chemical, such as a physical stressor,
environmental stressor, or a mechanical force (e.g., increased
pressure or light exposure) (see, e.g., US 2005-0059148).
[1005] The retinal cell stressor model system may also include a
cell stressor such as, but not limited to, a stressor that may be a
risk factor in a disease or disorder or that may contribute to the
development or progression of a disease or disorder, including but
not limited to, light of varying wavelengths and intensities; A2E;
cigarette smoke condensate exposure; oxidative stress (e.g., stress
related to the presence of or exposure to hydrogen peroxide,
nitroprusside, Zn++, or Fe++); increased pressure (e.g.,
atmospheric pressure or hydrostatic pressure), glutamate or
glutamate agonist (e.g., N-methyl-D-aspartate (NMDA);
alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionate (AMPA);
kainic acid; quisqualic acid; ibotenic acid; quinolinic acid;
aspartate; trans-1-aminocyclopentyl-1,3-dicarboxylate (ACPD));
amino acids (e.g., aspartate, L-cysteine;
beta-N-methylamine-L-alanine); heavy metals (such as lead); various
toxins (for example, mitochondrial toxins (e.g., malonate,
3-nitroproprionic acid; rotenone, cyanide); MPTP
(1-methyl-4-phenyl-1,2,3,6,-tetrahydropyridine), which metabolizes
to its active, toxic metabolite MPP+ (1-methyl-4-phenylpryidine));
6-hydroxydopamine; alpha-synuclein; protein kinase C activators
(e.g., phorbol myristate acetate); biogenic amino stimulants (for
example, methamphetamine, MDMA (3-4
methylenedioxymethamphetamine)); or a combination of one or more
stressors. Useful retinal cell stressors include those that mimic a
neurodegenerative disease that affects any one or more of the
mature retinal cells described herein. A chronic disease model is
of particular importance because most neurodegenerative diseases
are chronic. Through use of this in vitro cell culture system, the
earliest events in long-term disease development processes may be
identified because an extended period of time is available for
cellular analysis.
[1006] A retinal cell stressor may alter (i.e., increase or
decrease in a statistically significant manner) viability of
retinal cells such as by altering survival of retinal cells,
including retinal neuronal cells and RPE cells, or by altering
neurodegeneration of retinal neuronal cells and/or RPE cells.
Preferably, a retinal cell stressor adversely affects a retinal
neuronal cell or RPE cell such that survival of a retinal neuronal
cell or RPE cell is decreased or adversely affected (i.e., the
length of time during which the cells are viable is decreased in
the presence of the stressor) or neurodegeneration (or neuron cell
injury) of the cell is increased or enhanced. The stressor may
affect only a single retinal cell type in the retinal cell culture
or the stressor may affect two, three, four, or more of the
different cell types. For example, a stressor may alter viability
and survival of photoreceptor cells but not affect all the other
major cell types (e.g., ganglion cells, amacrine cells, horizontal
cells, bipolar cells, RPE, and Muller glia). Stressors may shorten
the survival time of a retinal cell (in vivo or in vitro), increase
the rapidity or extent of neurodegeneration of a retinal cell, or
in some other manner adversely affect the viability, morphology,
maturity, or lifespan of the retinal cell.
[1007] The effect of a cell stressor (in the presence and absence
of a compound described herein) on the viability of retinal cells
in the cell culture system may be determined for one or more of the
different retinal cell types. Determination of cell viability may
include evaluating structure and/or a function of a retinal cell
continually at intervals over a length of time or at a particular
time point after the retinal cell culture is prepared. Viability or
long term survival of one or more different retinal cell types or
one or more different retinal neuronal cell types may be examined
according to one or more biochemical or biological parameters that
are indicative of reduced viability, such as apoptosis or a
decrease in a metabolic function, prior to observation of a
morphological or structural alteration.
[1008] A chemical, biological, or physical cell stressor may reduce
viability of one or more of the retinal cell types present in the
cell culture system when the stressor is added to the cell culture
under conditions described herein for maintaining the long-term
cell culture. Alternatively, one or more culture conditions may be
adjusted so that the effect of the stressor on the retinal cells
can be more readily observed. For example, the concentration or
percent of fetal bovine serum may be reduced or eliminated from the
cell culture when cells are exposed to a particular cell stressor
(see, e.g., US 2005-0059148). Alternatively, retinal cells cultured
in media containing serum at a particular concentration for
maintenance of the cells may be abruptly exposed to media that does
not contain any level of serum.
[1009] The retinal cell culture may be exposed to a cell stressor
for a period of time that is determined to reduce the viability of
one or more retinal cell types in the retinal cell culture system.
The cells may be exposed to a cell stressor immediately upon
plating of the retinal cells after isolation from retinal tissue.
Alternatively, the retinal cell culture may be exposed to a
stressor after the culture is established, or any time thereafter.
When two or more cell stressors are included in the retinal cell
culture system, each stressor may be added to the cell culture
system concurrently and for the same length of time or may be added
separately at different time points for the same length of time or
for differing lengths of time during the culturing of the retinal
cell system. A compound described herein may be added before the
retinal cell culture is exposed to a cell stressor, may be added
concurrently with the cell stressor, or may be added after exposure
of the retinal cell culture to the stressor.
[1010] Photoreceptors may be identified using antibodies that
specifically bind to photoreceptor-specific proteins such as
opsins, peripherins, and the like. Photoreceptors in cell culture
may also be identified as a morphologic subset of
immunocytochemically labeled cells by using a pan-neuronal marker
or may be identified morphologically in enhanced contrast images of
live cultures. Outer segments can be detected morphologically as
attachments to photoreceptors.
[1011] Retinal cells including photoreceptors can also be detected
by functional analysis. For example, electrophysiology methods and
techniques may be used for measuring the response of photoreceptors
to light. Photoreceptors exhibit specific kinetics in a graded
response to light. Calcium-sensitive dyes may also be used to
detect graded responses to light within cultures containing active
photoreceptors. For analyzing stress-inducing compounds or
potential neurotherapeutics, retinal cell cultures can be processed
for immunocytochemistry, and photoreceptors and/or other retinal
cells can be counted manually or by computer software using
photomicroscopy and imaging techniques. Other immunoassays known in
the art (e.g., ELISA, immunoblotting, flow cytometry) may also be
useful for identifying and characterizing the retinal cells and
retinal neuronal cells of the cell culture model system described
herein.
[1012] The retinal cell culture stress models may also be useful
for identification of both direct and indirect pharmacologic agent
effects by the bioactive agent of interest, such as a compound as
described herein. For example, a bioactive agent added to the cell
culture system in the presence of one or more retinal cell
stressors may stimulate one cell type in a manner that enhances or
decreases the survival of other cell types. Cell/cell interactions
and cell/extracellular component interactions may be important in
understanding mechanisms of disease and drug function. For example,
one neuronal cell type may secrete trophic factors that affect
growth or survival of another neuronal cell type (see, e.g., WO
99/29279).
[1013] In another embodiment, a compound described herein is
incorporated into screening assays comprising the retinal cell
culture stress model system described herein to determine whether
and/or to what level or degree the compound increases or prolongs
viability (i.e., increases in a statistically significant or
biologically significant manner) of a plurality of retinal cells. A
person skilled in the art would readily appreciate and understand
that as described herein a retinal cell that exhibits increased
viability means that the length of time that a retinal cell
survives in the cell culture system is increased (increased
lifespan) and/or that the retinal cell maintains a biological or
biochemical function (normal metabolism and organelle function;
lack of apoptosis; etc.) compared with a retinal cell cultured in
an appropriate control cell system (e.g., the cell culture system
described herein in the absence of the compound). Increased
viability of a retinal cell may be indicated by delayed cell death
or a reduced number of dead or dying cells; maintenance of
structure and/or morphology; lack of or delayed initiation of
apoptosis; delay, inhibition, slowed progression, and/or abrogation
of retinal neuronal cell neurodegeneration or delaying or
abrogating or preventing the effects of neuronal cell injury.
Methods and techniques for determining viability of a retinal cell
and thus whether a retinal cell exhibits increased viability are
described in greater detail herein and are known to persons skilled
in the art.
[1014] In certain embodiments, a method is provided for determining
whether a compound described herein, enhances survival of
photoreceptor cells. One method comprises contacting a retinal cell
culture system as described herein with a compound described herein
under conditions and for a time sufficient to permit interaction
between the retinal neuronal cells and the compound. Enhanced
survival (prolonged survival) may be measured according to methods
described herein and known in the art, including detecting
expression of rhodopsin.
[1015] The capability of a compound described herein to increase
retinal cell viability and/or to enhance, promote, or prolong cell
survival (that is, to extend the time period in which retinal
cells, including retinal neuronal cells, are viable), and/or
impair, inhibit, or impede degeneration as a direct or indirect
result of the herein described stress may be determined by any one
of several methods known to those skilled in the art. For example,
changes in cell morphology in the absence and presence of the
compound may be determined by visual inspection such as by light
microscopy, confocal microscopy, or other microscopy methods known
in the art. Survival of cells can also be determined by counting
viable and/or nonviable cells, for instance. Immunochemical or
immunohistological techniques (such as fixed cell staining or flow
cytometry) may be used to identify and evaluate cytoskeletal
structure (e.g., by using antibodies specific for cytoskeletal
proteins such as glial fibrillary acidic protein, fibronectin,
actin, vimentin, tubulin, or the like) or to evaluate expression of
cell markers as described herein. The effect of a compound
described herein on cell integrity, morphology, and/or survival may
also be determined by measuring the phosphorylation state of
neuronal cell polypeptides, for example, cytoskeletal polypeptides
(see, e.g., Sharma et al., J. Biol. Chem. 274:9600-06 (1999); Li et
al., J. Neurosci. 20:6055-62 (2000)). Cell survival or,
alternatively cell death, may also be determined according to
methods described herein and known in the art for measuring
apoptosis (for example, annexin V binding, DNA fragmentation
assays, caspase activation, marker analysis, e.g., poly(ADP-ribose)
polymerase (PARP), etc.).
[1016] In the vertebrate eye, for example, a mammalian eye, the
formation of A2E is a light-dependent process and its accumulation
leads to a number of negative effects in the eye. These include
destabilization of retinal pigment epithelium (RPE) membranes,
sensitization of cells to blue-light damage, and impaired
degradation of phospholipids. Products of the oxidation of A2E (and
A2E related molecules) by molecular oxygen (oxiranes) were shown to
induce DNA damage in cultured RPE cells. All these factors lead to
a gradual decrease in visual acuity and eventually to vision loss.
If reducing the formation of retinals during vision processes were
possible, this reduction would lead to decreased amounts of A2E in
the eye. Without wishing to be bound by theory, decreased
accumulation of A2E may reduce or delay degenerative processes in
the RPE and retina and thus may slow down or prevent vision loss in
dry AMD and Stargardt's Disease.
[1017] In another embodiment, methods are provided for treating
and/or preventing degenerative diseases and disorders, including
neurodegenerative retinal diseases and ophthalmic diseases, and
retinal diseases and disorders as described herein. A subject in
need of such treatment may be a human or non-human primate or other
animal who has developed symptoms of a degenerative retinal disease
or who is at risk for developing a degenerative retinal disease. As
described herein a method is provided for treating (which includes
preventing or prophylaxis) an ophthalmic disease or disorder by
administrating to a subject a composition comprising a
pharmaceutically acceptable carrier and a compound described herein
(e.g., a compound having the structure of Formula (I), (II), (IIa),
(III), (IIIa), (IV), or (IVa), and substructures thereof.) As
described herein, a method is provided for enhancing survival of
neuronal cells such as retinal neuronal cells, including
photoreceptor cells, and/or inhibiting degeneration of retinal
neuronal cells by administering the pharmaceutical compositions
described herein comprising a compound described herein.
[1018] Enhanced survival (or prolonged or extended survival) of one
or more retinal cell types in the presence of a compound described
herein indicates that the compound may be an effective agent for
treatment of a degenerative disease, particularly a retinal disease
or disorder, and including a neurodegenerative retinal disease or
disorder. Cell survival and enhanced cell survival may be
determined according to methods described herein and known to a
skilled artisan including viability assays and assays for detecting
expression of retinal cell marker proteins. For determining
enhanced survival of photoreceptor cells, opsins may be detected,
for instance, including the protein rhodopsin that is expressed by
rods.
[1019] In another embodiment, the subject is being treated for
Stargardt's disease or Stargardt's macular degeneration. In
Stargardt's disease, which is associated with mutations in the
ABCA4 (also called ABCR) transporter, the accumulation of
all-trans-retinal has been proposed to be responsible for the
formation of a lipofuscin pigment, A2E, which is toxic towards
retinal cells and causes retinal degeneration and consequently loss
of vision.
[1020] In yet another embodiment, the subject is being treated for
age-related macular degeneration (AMD). In various embodiments, AMD
can be wet- or dry-form. In AMD, vision loss primarily occurs when
complications late in the disease either cause new blood vessels to
grow under the macula or the macula atrophies. Without intending to
be bound by any particular theory, the accumulation of
all-trans-retinal has been proposed to be responsible for the
formation of a lipofuscin pigment,
N-retinylidene-N-retinylethanolamine (A2E) and A2E related
molecules, which are toxic towards RPE and retinal cells and cause
retinal degeneration and consequently loss of vision.
[1021] A neurodegenerative retinal disease or disorder for which
the compounds and methods described herein may be used for
treating, curing, preventing, ameliorating the symptoms of, or
slowing, inhibiting, or stopping the progression of, is a disease
or disorder that leads to or is characterized by retinal neuronal
cell loss, which is the cause of visual impairment. Such a disease
or disorder includes but is not limited to age-related macular
degeneration (including dry-form and wet-form of macular
degeneration) and Stargardt's macular dystrophy.
[1022] Age-related macular degeneration as described herein is a
disorder that affects the macula (central region of the retina) and
results in the decline and loss of central vision. Age-related
macular degeneration occurs typically in individuals over the age
of 55 years. The etiology of age-related macular degeneration may
include both environmental influences and genetic components (see,
e.g., Lyengar et al., Am. J. Hum. Genet. 74:20-39 (2004) (Epub 2003
Dec. 19); Kenealy et al., Mol. Vis. 10:57-61 (2004); Gorin et al.,
Mol. Vis. 5:29 (1999)). More rarely, macular degeneration occurs in
younger individuals, including children and infants, and generally,
these disorders results from a genetic mutation. Types of juvenile
macular degeneration include Stargardt's disease (see, e.g., Glazer
et al., Ophthalmol. Clin. North Am. 15:93-100, viii (2002); Weng et
al., Cell 98:13-23 (1999)); Doyne's honeycomb retinal dystrophy
(see, e.g., Kermani et al., Hum. Genet. 104:77-82 (1999)); Sorsby's
fundus dystrophy, Malattia Levintinese, fundus flavimaculatus, and
autosomal dominant hemorrhagic macular dystrophy (see also Seddon
et al., Ophthalmology 108:2060-67 (2001); Yates et al., J. Med.
Genet. 37:83-7 (2000); Jaakson et al., Hum. Mutat. 22:395-403
(2003)). Geographic atrophy of the RPE is an advanced form of
non-neovascular dry-type age-related macular degeneration, and is
associated with atrophy of the choriocapillaris, RPE, and
retina.
[1023] Stargardt's macular degeneration, a recessive inherited
disease, is an inherited blinding disease of children. The primary
pathologic defect in Stargardt's disease is also an accumulation of
toxic lipofuscin pigments such as A2E in cells of the retinal
pigment epithelium (RPE). This accumulation appears to be
responsible for the photoreceptor death and severe visual loss
found in Stargardt's patients. The compounds described herein may
slow the synthesis of 11-cis-retinaldehyde (11cRAL or retinal) and
regeneration of rhodopsin by inhibiting isomerase in the visual
cycle. Light activation of rhodopsin results in its release of
all-trans-retinal, which constitutes the first reactant in A2E
biosynthesis. Treatment with a compound described herein may
inhibit lipofuscin accumulation and thus delay the onset of visual
loss in Stargardt's and AMD patients without toxic effects that
would preclude treatment with a compound described herein. The
compounds described herein may be used for effective treatment of
other forms of retinal or macular degeneration associated with
lipofuscin accumulation.
[1024] Administration of a compound described herein to a subject
can prevent formation of the lipofuscin pigment, A2E (and A2E
related molecules), that is toxic towards retinal cells and causes
retinal degeneration. In certain embodiments, administration of a
compound described herein can lessen the production of waste
products, e.g., lipofuscin pigment, A2E (and A2E related
molecules), ameliorate the development of AMD (e.g., dry-form) and
Stargardt's disease, and reduce or slow vision loss (e.g.,
choroidal neovascularization and/or chorioretinal atrophy). In
previous studies, with 13-cis-retinoic acid (Accutane.RTM. or
Isotretinoin), a drug commonly used for the treatment of acne and
an inhibitor of 11-cis-retinol dehydrogenase, has been administered
to patients to prevent A2E accumulation in the RPE. However, a
major drawback in this proposed treatment is that 13-cis-retinoic
acid can easily isomerize to all-trans-retinoic acid.
All-trans-retinoic acid is a very potent teratogenic compound that
adversely affects cell proliferation and development. Retinoic acid
also accumulates in the liver and may be a contributing factor in
liver diseases.
[1025] In yet other embodiments, a compound described herein is
administered to a subject such as a human with a mutation in the
ABCA4 transporter in the eye. The compound described herein can
also be administered to an aging subject. As used herein, an aging
human subject is typically at least 45, or at least 50, or at least
60, or at least 65 years old. In Stargardt's disease, which is
associated with mutations in the ABCA4 transporter, the
accumulation of all-trans-retinal has been proposed to be
responsible for the formation of a lipofuscin pigment, A2E (and A2E
related molecules), that is toxic towards retinal cells and causes
retinal degeneration and consequently loss of vision. Without
wishing to be bound by theory, a compound described herein may be a
strong inhibitor of an isomerase involved in the visual cycle.
Treating patients with a compound as described herein may prevent
or slow the formation of A2E (and A2E related molecules) and can
have protective properties for normal vision.
[1026] In other certain embodiments, one or more of the compounds
described herein may be used for treating other ophthalmic diseases
or disorders, for example, glaucoma, retinal detachment,
hemorrhagic retinopathy, retinitis pigmentosa, an inflammatory
retinal disease, proliferative vitreoretinopathy, retinal
dystrophy, hereditary optic neuropathy, Sorsby's fundus dystrophy,
uveitis, a retinal injury, optical neuropathy, and retinal
disorders associated with other neurodegenerative diseases such as
Alzheimer's disease, multiple sclerosis, Parkinson's disease or
other neurodegenerative diseases that affect brain cells, a retinal
disorder associated with viral infection, or other conditions such
as AIDS. A retinal disorder also includes light damage to the
retina that is related to increased light exposure (i.e.,
overexposure to light), for example, accidental strong or intense
light exposure during surgery; strong, intense, or prolonged
sunlight exposure, such as at a desert or snow covered terrain;
during combat, for example, when observing a flare or explosion or
from a laser device, and the like. Retinal diseases can be of
degenerative or non-degenerative nature. Non-limiting examples of
degenerative retinal diseases include age-related macular
degeneration, and Stargardt's macular dystrophy. Examples of
non-degenerative retinal diseases include but are not limited
hemorrhagic retinopathy, retinitis pigmentosa, optic neuropathy,
inflammatory retinal disease, diabetic retinopathy, diabetic
maculopathy, retinal blood vessel occlusion, retinopathy of
prematurity, or ischemia reperfusion related retinal injury,
proliferative vitreoretinopathy, retinal dystrophy, hereditary
optic neuropathy, Sorsby's fundus dystrophy, uveitis, a retinal
injury, a retinal disorder associated with Alzheimer's disease, a
retinal disorder associated with multiple sclerosis, a retinal
disorder associated with Parkinson's disease, a retinal disorder
associated with viral infection, a retinal disorder related to
light overexposure, and a retinal disorder associated with
AIDS.
[1027] In other certain embodiments, at least one of the compounds
described herein may be used for treating, curing, preventing,
ameliorating the symptoms of, or slowing, inhibiting, or stopping
the progression of, certain ophthalmic diseases and disorders
including but not limited to diabetic retinopathy, diabetic
maculopathy, diabetic macular edema, retinal ischemia,
ischemia-reperfusion related retinal injury, and retinal blood
vessel occlusion (including venous occlusion and arterial
occlusion).
[1028] Diabetic retinopathy is a leading cause of blindness in
humans and is a complication of diabetes. Diabetic retinopathy
occurs when diabetes damages blood vessels inside the retina.
Non-proliferative retinopathy is a common, usually mild form that
generally does not interfere with vision. Abnormalities are limited
to the retina, and vision is impaired only if the macula is
involved. If left untreated retinopathy can progress to
proliferative retinopathy, the more serious form of diabetic
retinopathy. Proliferative retinopathy occurs when new blood
vessels proliferate in and around the retina. Consequently,
bleeding into the vitreous, swelling of the retina, and/or retinal
detachment may occur, leading to blindness.
[1029] Other ophthalmic diseases and disorders that may be treated
using the methods and compositions described herein include
diseases, disorders, and conditions that are associated with,
exacerbated by, or caused by ischemia in the retina. Retinal
ischemia includes ischemia of the inner retina and the outer
retina. Retinal ischemia can occur from either choroidal or retinal
vascular diseases, such as central or branch retinal vision
occlusion, collagen vascular diseases and thrombocytopenic purpura.
Retinal vasculitis and occlusion is seen with Eales disease and
systemic lupus erythematosus.
[1030] Retinal ischemia may be associated with retinal blood vessel
occlusion. In the United States, both branch and central retinal
vein occlusions are the second most common retinal vascular
diseases after diabetic retinopathy. About 7% to 10% of patients
who have retinal venous occlusive disease in one eye eventually
have bilateral disease. Visual field loss commonly occurs from
macular edema, ischemia, or vitreous hemorrhage secondary to disc
or retinal neovascularization induced by the release of vascular
endothelial growth factor.
[1031] Arteriolosclerosis at sites of retinal arteriovenous
crossings (areas in which arteries and veins share a common
adventitial sheath) causes constriction of the wall of a retinal
vein by a crossing artery. The constriction results in thrombus
formation and subsequent occlusion of the vein. The blocked vein
may lead to macular edema and hemorrhage secondary to breakdown in
the blood-retina barrier in the area drained by the vein,
disruption of circulation with turbulence in venous flow,
endothelial damage, and ischemia. Clinically, areas of ischemic
retina appear as feathery white patches called cotton-wool
spots.
[1032] Branch retinal vein occlusions with abundant ischemia cause
acute central and paracentral visual field loss corresponding to
the location of the involved retinal quadrants. Retinal
neovascularization due to ischemia may lead to vitreous hemorrhage
and subacute or acute vision loss.
[1033] Two types of central retinal vein occlusion, ischemic and
nonischemic, may occur depending on whether widespread retinal
ischemia is present. Even in the nonischemic type, the macula may
still be ischemic. Approximately 25% central retinal vein occlusion
is ischemic. Diagnosis of central retinal vein occlusion can
usually be made on the basis of characteristic ophthalmoscopic
findings, including retinal hemorrhage in all quadrants, dilated
and tortuous veins, and cotton-wool spots. Macular edema and foveal
ischemia can lead to vision loss. Extracellular fluid increases
interstitial pressure, which may result in areas of retinal
capillary closure (i.e., patchy ischemic retinal whitening) or
occlusion of a cilioretinal artery.
[1034] Patients with ischemic central retinal vein occlusion are
more likely to present with a sudden onset of vision loss and have
visual acuity of less than 20/200, a relative afferent pupillary
defect, abundant intraretinal hemorrhages, and extensive
nonperfusion on fluorescein angiography. The natural history of
ischemic central retinal vein occlusion is associated with poor
outcomes: eventually, approximately two-thirds of patients who have
ischemic central retinal vein occlusion will have ocular
neovascularization and one-third will have neovascular glaucoma.
The latter condition is a severe type of glaucoma that may lead to
rapid visual field and vision loss, epithelial edema of the cornea
with secondary epithelial erosion and predisposition to bacterial
keratitis, severe pain, nausea and vomiting, and, eventually,
phthisis bulbi (atrophy of the globe with no light perception).
[1035] As used herein, a patient (or subject) may be any mammal,
including a human, that may have or be afflicted with a
neurodegenerative disease or condition, including an ophthalmic
disease or disorder, or that may be free of detectable disease.
Accordingly, the treatment may be administered to a subject who has
an existing disease, or the treatment may be prophylactic,
administered to a subject who is at risk for developing the disease
or condition. Treating or treatment refers to any indicia of
success in the treatment or amelioration of an injury, pathology or
condition, including any objective or subjective parameter such as
abatement; remission; diminishing of symptoms or making the injury,
pathology, or condition more tolerable to the patient; slowing in
the rate of degeneration or decline; making the final point of
degeneration less debilitating; or improving a subject's physical
or mental well-being.
[1036] The treatment or amelioration of symptoms can be based on
objective or subjective parameters; including the results of a
physical examination. Accordingly, the term "treating" includes the
administration of the compounds or agents described herein to treat
pain, hyperalgesia, allodynia, or nociceptive events and to prevent
or delay, to alleviate, or to arrest or inhibit development of the
symptoms or conditions associated with pain, hyperalgesia,
allodynia, nociceptive events, or other disorders. The term
"therapeutic effect" refers to the reduction, elimination, or
prevention of the disease, symptoms of the disease, or sequelae of
the disease in the subject. Treatment also includes restoring or
improving retinal neuronal cell functions (including photoreceptor
function) in a vertebrate visual system, for example, such as
visual acuity and visual field testing etc., as measured over time
(e.g., as measured in weeks or months). Treatment also includes
stabilizing disease progression (i.e., slowing, minimizing, or
halting the progression of an ophthalmic disease and associated
symptoms) and minimizing additional degeneration of a vertebrate
visual system. Treatment also includes prophylaxis and refers to
the administration of a compound described herein to a subject to
prevent degeneration or further degeneration or deterioration or
further deterioration of the vertebrate visual system of the
subject and to prevent or inhibit development of the disease and/or
related symptoms and sequelae.
[1037] Various methods and techniques practiced by a person skilled
in the medical and ophthalmological arts to determine and evaluate
a disease state and/or to monitor and assess a therapeutic regimen
include, for example, fluorescein angiogram, fundus photography,
indocyanine green dye tracking of the choroidal circulatory system,
opthalmoscopy, optical coherence tomography (OCT), and visual
acuity testing.
[1038] A fluorescein angiogram involves injecting a fluorescein dye
intravenously and then observing any leakage of the dye as it
circulates through the eye. Intravenous injection of indocyanine
green dye may also be used to determine if vessels in the eye are
compromised, particularly in the choroidal circulatory system that
is just behind the retina. Fundus photography may be used for
examining the optic nerve, macula, blood vessels, retina, and the
vitreous. Microaneurysms are visible lesions in diabetic
retinopathy that may be detected in digital fundus images early in
the disease (see, e.g., U.S. Patent Application Publication No.
2007/0002275). An ophthalmoscope may be used to examine the retina
and vitreous. Opthalmoscopy is usually performed with dilated
pupils, to allow the best view inside the eye. Two types of
ophthalmoscopes may be used: direct and indirect. The direct
ophthalmoscope is generally used to view the optic nerve and the
central retina. The periphery, or entire retina, may be viewed by
using an indirect ophthalmoscope. Optical coherence tomography
(OCT) produces high resolution, high speed, non-invasive,
cross-sectional images of body tissue. OCT is noninvasive and
provides detection of microscopic early signs of disruption in
tissues.
[1039] A subject or patient refers to any vertebrate or mammalian
patient or subject to whom the compositions described herein can be
administered. The term "vertebrate" or "mammal" includes humans and
non-human primates, as well as experimental animals such as
rabbits, rats, and mice, and other animals, such as domestic pets
(such as cats, dogs, horses), farm animals, and zoo animals.
Subjects in need of treatment using the methods described herein
may be identified according to accepted screening methods in the
medical art that are employed to determine risk factors or symptoms
associated with an ophthalmic disease or condition described herein
or to determine the status of an existing ophthalmic disease or
condition in a subject. These and other routine methods allow the
clinician to select patients in need of therapy using the methods
and formulations described herein.
III. Pharmaceutical Compositions
[1040] In certain embodiments, a compound described herein may be
administered as a pure chemical. In other embodiments, the compound
described herein can be combined with a pharmaceutical carrier
(also referred to herein as a pharmaceutically acceptable excipient
(i.e., a pharmaceutically suitable and acceptable carrier, diluent,
etc., which is a non-toxic, inert material that does not interfere
with the activity of the active ingredient)) selected on the basis
of a chosen route of administration and standard pharmaceutical
practice as described, for example, in Remington: The Science and
Practice of Pharmacy (Gennaro, 21.sup.st Ed. Mack Pub. Co., Easton,
Pa. (2005)), the disclosure of which is hereby incorporated herein
by reference, in its entirety.
[1041] Accordingly, provided herein is a pharmaceutical composition
comprising one or more compounds described herein, or a
stereoisomer, tautomer, prodrug, pharmaceutically acceptable salt,
hydrate, solvate, acid salt hydrate, N-oxide or isomorphic
crystalline form thereof, of a compound described herein, together
with one or more pharmaceutically acceptable carriers and,
optionally, other therapeutic and/or prophylactic ingredients. The
carrier(s) (or excipient(s)) is acceptable or suitable if the
carrier is compatible with the other ingredients of the composition
and not deleterious to the recipient (i.e., the subject) of the
composition. A pharmaceutically acceptable or suitable composition
includes an ophthalmologically suitable or acceptable
composition.
[1042] Thus, another embodiment provides a pharmaceutical
composition comprising a pharmaceutically acceptable excipient and
a compound having a structure of Formula (I) or tautomer,
stereoisomer, geometric isomer or a pharmaceutically acceptable
solvate, hydrate, salt, N-oxide or prodrug thereof:
##STR00287##
wherein, [1043] Z is a bond, --C(R.sup.1)(R.sup.2)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--,
--X--C(R.sup.31)(R.sup.32)--,
--C(R.sup.9)(R.sup.10)--C(R.sup.1)(R.sup.2)--C(R.sup.36)(R.sup.37)--,
--C(R.sup.38)(R.sup.39)--X--C(R.sup.31)(R.sup.32)--, or
--X--C(R.sup.31)(R.sup.32)--C(R.sup.1)(R.sup.2)--; [1044] X is
--O--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --N(R.sup.30)--,
--C(.dbd.O)--, --C(.dbd.CH.sub.2)--, --C(.dbd.N--NR.sup.35)--, or
--C(.dbd.N--OR.sup.35)--; [1045] G is selected from
--N(R.sup.42)--SO.sub.2--R.sup.40,
--N(R.sup.42)C(.dbd.O)--R.sup.40,
--N(R.sup.42)C(.dbd.O)--OR.sup.40,
--N(R.sup.42)--C(R.sup.42)(R.sup.42)--R.sup.40,
--N(R.sup.42)--C(.dbd.O)--N(R.sup.43)(R.sup.43), or
--N(R.sup.42)--C(.dbd.S)--N(R.sup.43)(R.sup.43); [1046] R.sup.40 is
selected from --C(R.sup.16)(R.sup.17)(R.sup.18), aryl, or
heteroaryl; [1047] each R.sup.42 is independently selected from
hydrogen, alkyl or aryl; [1048] each R.sup.43 is independently
selected from hydrogen, alkyl, cycloalkyl, aralkyl, alkenyl,
alkynyl, C-attached heterocyclyl, aryl, or heteroaryl; or two
R.sup.43 groups, together with the nitrogen to which they are
attached, may form a heterocyclyl; [1049] R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halogen, C.sub.1-C.sub.5
alkyl, fluoroalkyl, --OR.sup.6 or --NR.sup.7R.sup.8; or R.sup.1 and
R.sup.2 together form an oxo; [1050] R.sup.31 and R.sup.32 are each
independently selected from hydrogen, C.sub.1-C.sub.5 alkyl, or
fluoroalkyl; [1051] R.sup.38 and R.sup.39 are each independently
selected from hydrogen, C.sub.1-C.sub.5 alkyl, or fluoroalkyl;
R.sup.36 and R.sup.37 are each independently selected from
hydrogen, halogen, C.sub.1-C.sub.5 alkyl, fluoroalkyl, --OR.sup.6
or --NR.sup.7R.sup.8; or R.sup.36 and R.sup.37 together form an
oxo; or optionally, R.sup.36 and R.sup.1 together form a direct
bond to provide a double bond; or optionally, R.sup.36 and R.sup.1
together form a direct bond, and R.sup.37 and R.sup.2 together form
a direct bond to provide a triple bond; [1052] R.sup.3 and R.sup.4
are each independently selected from hydrogen, alkyl, alkenyl,
fluoroalkyl, aryl, heteroaryl, carbocyclyl or C-attached
heterocyclyl; or R.sup.3 and R.sup.4 together with the carbon atom
to which they are attached, form a carbocyclyl or heterocyclyl; or
R.sup.3 and R.sup.4 together form an imino; [1053] R.sup.7 and
R.sup.8 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.13, SO.sub.2R.sup.13,
CO.sub.2R.sup.13 or SO.sub.2NR.sup.24R.sup.25; or R.sup.7 and
R.sup.8 together with the nitrogen atom to which they are attached,
form an N-heterocyclyl; [1054] R.sup.9 and R.sup.10 are each
independently selected from hydrogen, halogen, alkyl, fluoroalkyl,
--OR.sup.19, --NR.sup.20R.sup.21 or carbocyclyl; or R.sup.9 and
R.sup.10 form an oxo; or optionally, R.sup.9 and R.sup.1 together
form a direct bond to provide a double bond; or optionally, R.sup.9
and R.sup.1 together form a direct bond, and R.sup.10 and R.sup.2
together form a direct bond to provide a triple bond; [1055]
R.sup.11 and R.sup.12 are each independently selected from
hydrogen, alkyl, carbocyclyl, --C(.dbd.O)R.sup.23, --C(NH)NH.sub.2,
SO.sub.2R.sup.23, CO.sub.2R.sup.23 or SO.sub.2NR.sup.28R.sup.29; or
R.sup.11 and R.sup.12, together with the nitrogen atom to which
they are attached, form an N-heterocyclyl; [1056] each R.sup.13,
R.sup.22 and R.sup.23 is independently selected from alkyl,
heteroalkyl, alkenyl, aryl, aralkyl, carbocyclyl, heteroaryl or
heterocyclyl; [1057] R.sup.6, R.sup.19, R.sup.30, R.sup.34 and
R.sup.35 are each independently hydrogen or alkyl; [1058] R.sup.20
and R.sup.21 are each independently selected from hydrogen, alkyl,
carbocyclyl, heterocyclyl, --C(.dbd.O)R.sup.22, SO.sub.2R.sup.22,
CO.sub.2R.sup.22 or SO.sub.2NR.sup.26R.sup.27; or R.sup.20 and
R.sup.21 together with the nitrogen atom to which they are
attached, form an N-heterocyclyl; and [1059] each R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28, and R.sup.29 is
independently selected from hydrogen, alkyl, alkenyl, fluoroalkyl,
aryl, heteroaryl, carbocyclyl or heterocyclyl; [1060] R.sup.16 and
R.sup.17 are each independently selected from hydrogen, alkyl,
halo, aryl, heteroaryl, aralkyl, heteroaryalkyl or fluoroalkyl; or
R.sup.16 and R.sup.17, together with the carbon to which they are
attached form a carbocyclyl or heterocycle; [1061] R.sup.18 is
selected from hydrogen, alkyl, alkoxy, hydroxy, halo or
fluoroalkyl; [1062] each R.sup.33 is independently selected from
halogen, OR.sup.34, alkyl, or fluoroalkyl; and n is 0, 1, 2, 3, or
4.
[1063] One embodiment provides a pharmaceutical composition
comprising a pharmaceutically acceptable excipient and a compound
of Formula (II), (IIa), (III), (IIIa), (IV), or (IVa) as described
herein, or tautomer, stereoisomer, geometric isomer or a
pharmaceutically acceptable solvate, hydrate, salt, N-oxide or
prodrug thereof.
[1064] A pharmaceutical composition (e.g., for oral administration
or delivery by injection, or combined devices, or for application
as an eye drop) may be in the form of a liquid or solid. A liquid
pharmaceutical composition may include, for example, one or more of
the following: sterile diluents such as water for injection, saline
solution, preferably physiological saline, Ringer's solution,
isotonic sodium chloride, fixed oils that may serve as the solvent
or suspending medium, polyethylene glycols, glycerin, propylene
glycol or other solvents; antibacterial agents; antioxidants;
chelating agents; buffers and agents for the adjustment of tonicity
such as sodium chloride or dextrose. A parenteral preparation can
be enclosed in ampules, disposable syringes or multiple dose vials
made of glass or plastic. Physiological saline is commonly used as
an excipient, and an injectable pharmaceutical composition or a
composition that is delivered ocularly is preferably sterile.
[1065] At least one compound described herein can be administered
to human or other nonhuman vertebrates. In certain embodiments, the
compound is substantially pure, in that it contains less than about
5% or less than about 1%, or less than about 0.1%, of other organic
small molecules, such as contaminating intermediates or by-products
that are created, for example, in one or more of the steps of a
synthesis method. In other embodiments, a combination of one or
more compounds described herein can be administered.
[1066] A compound described herein can be delivered to a subject by
any suitable means, including, for example, orally, parenterally,
intraocularly, intravenously, intraperitoneally, intranasally (or
other delivery methods to the mucous membranes, for example, of the
nose, throat, and bronchial tubes), or by local administration to
the eye, or by an intraocular or periocular device. Modes of local
administration can include, for example, eye drops, intraocular
injection or periocular injection. Periocular injection typically
involves injection of the synthetic isomerization inhibitor, i.e.,
compound as described herein, under the conjunctiva or into the
Tennon's space (beneath the fibrous tissue overlying the eye).
Intraocular injection typically involves injection of the compound
described herein into the vitreous. In certain embodiments, the
administration is non-invasive, such as by eye drops or oral dosage
form, or as a combined device.
[1067] A compound described herein can be formulated for
administration using pharmaceutically acceptable (suitable)
carriers or vehicles as well as techniques routinely used in the
art. A pharmaceutically acceptable or suitable carrier includes an
ophthalmologically suitable or acceptable carrier. A carrier is
selected according to the solubility of the compound described
herein. Suitable ophthalmological compositions include those that
are administrable locally to the eye, such as by eye drops,
injection or the like. In the case of eye drops, the formulation
can also optionally include, for example, ophthalmologically
compatible agents such as isotonizing agents such as sodium
chloride, concentrated glycerin, and the like; buffering agents
such as sodium phosphate, sodium acetate, and the like; surfactants
such as polyoxyethylene sorbitan mono-oleate (also referred to as
Polysorbate 80), polyoxyl stearate 40, polyoxyethylene hydrogenated
castor oil, and the like; stabilization agents such as sodium
citrate, sodium edentate, and the like; preservatives such as
benzalkonium chloride, parabens, and the like; and other
ingredients. Preservatives can be employed, for example, at a level
of from about 0.001 to about 1.0% weight/volume. The pH of the
formulation is usually within the range acceptable to
ophthalmologic formulations, such as within the range of about pH 4
to 8, or pH 5 to 7, or pH 6 to 7, or pH 4 to 7, or pH 5 to 8, or pH
6 to 8, or pH 4 to 6, or pH 5 to 6, or pH 7 to 8.
[1068] In additional embodiments, the compositions described herein
further comprise cyclodextrins. Cyclodextrins are cyclic
oligosaccharides containing 6, 7, or 8 glucopyranose units,
referred to as .alpha.-cyclodextrin, .beta.-cyclodextrin, or
.gamma.-cyclodextrin respectively. Cyclodextrins have been found to
be particularly useful in pharmaceutical formulations.
Cyclodextrins have a hydrophilic exterior, which enhances
water-soluble, and a hydrophobic interior which forms a cavity. In
an aqueous environment, hydrophobic portions of other molecules
often enter the hydrophobic cavity of cyclodextrin to form
inclusion compounds. Additionally, cyclodextrins are also capable
of other types of nonbonding interactions with molecules that are
not inside the hydrophobic cavity. Cyclodextrins have three free
hydroxyl groups for each glucopyranose unit, or 18 hydroxyl groups
on .alpha.-cyclodextrin, 21 hydroxyl groups on .beta.-cyclodextrin,
and 24 hydroxyl groups on .gamma.-cyclodextrin. One or more of
these hydroxyl groups can be reacted with any of a number of
reagents to form a large variety of cyclodextrin derivatives. Some
of the more common derivatives of cyclodextrin are hydroxypropyl
ethers, sulfonates, and sulfoalkylethers. Shown below is the
structure of .beta.-cyclodextrin and the
hydroxypropyl-.beta.-cyclodextrin (HP.beta.CD).
##STR00288##
[1069] The use of cyclodextrins in pharmaceutical compositions is
well known in the art as cyclodextrins and cyclodextrin derivatives
are often used to improve the solubility of a drug. Inclusion
compounds are involved in many cases of enhanced solubility;
however other interactions between cyclodextrins and insoluble
compounds can also improve solubility.
Hydroxypropyl-.beta.-cyclodextrin (HP.beta.CD) is commercially
available as a pyrogen free product. It is a nonhygroscopic white
powder that readily dissolves in water. HP.beta.CD is thermally
stable and does not degrade at neutral pH.
[1070] Ophthalmic formulations utilizing cyclodextrins have been
disclosed. For example, U.S. Pat. No. 5,227,372 discloses methods
related to retaining ophthalmological agents in ocular tissues. US
Patent Application Publication 2007/0149480 teaches the use of
cyclodextrins to prepare ophthalmic formulations of a small
molecule kinase inhibitor with poor water solubility.
[1071] The concentration of the cyclodextrin used in the
compositions and methods disclosed herein can vary according to the
physiochemical properties, pharmacokinetic properties, side effect
or adverse events, formulation considerations, or other factors
associated with the therapeutically active agent, or a salt or
prodrug thereof. The properties of other excipients in a
composition may also be important. Thus, the concentration or
amount of cyclodextrin used in accordance with the compositions and
methods disclosed herein can vary. In certain compositions, the
concentration of the cyclodextrin is from 10% to 25%.
[1072] For injection, the compound described herein can be provided
in an injection grade saline solution, in the form of an injectable
liposome solution, slow-release polymer system or the like.
Intraocular and periocular injections are known to those skilled in
the art and are described in numerous publications including, for
example, Spaeth, Ed., Ophthalmic Surgery: Principles of Practice,
W. B. Sanders Co., Philadelphia, Pa., 85-87, 1990.
[1073] For delivery of a composition comprising at least one of the
compounds described herein via a mucosal route, which includes
delivery to the nasal passages, throat, and airways, the
composition may be delivered in the form of an aerosol. The
compound may be in a liquid or powder form for intramucosal
delivery. For example, the composition may delivered via a
pressurized aerosol container with a suitable propellant, such as a
hydrocarbon propellant (e.g., propane, butane, isobutene). The
composition may be delivered via a non-pressurized delivery system
such as a nebulizer or atomizer.
[1074] Suitable oral dosage forms include, for example, tablets,
pills, sachets, or capsules of hard or soft gelatin,
methylcellulose or of another suitable material easily dissolved in
the digestive tract. Suitable nontoxic solid carriers can be used
which include, for example, pharmaceutical grades of mannitol,
lactose, starch, magnesium stearate, sodium saccharin, talcum,
cellulose, glucose, sucrose, magnesium carbonate, and the like.
(See, e.g., Remington: The Science and Practice of Pharmacy
(Gennaro, 21.sup.st Ed. Mack Pub. Co., Easton, Pa. (2005)).
[1075] The compounds described herein may be formulated for
sustained or slow-release. Such compositions may generally be
prepared using well known technology and administered by, for
example, oral, periocular, intraocular, rectal or subcutaneous
implantation, or by implantation at the desired target site.
Sustained-release formulations may contain an agent dispersed in a
carrier matrix and/or contained within a reservoir surrounded by a
rate controlling membrane. Excipients for use within such
formulations are biocompatible, and may also be biodegradable;
preferably the formulation provides a relatively constant level of
active component release. The amount of active compound contained
within a sustained-release formulation depends upon the site of
implantation, the rate and expected duration of release, and the
nature of the condition to be treated or prevented.
[1076] Systemic drug absorption of a drug or composition
administered via an ocular route is known to those skilled in the
art (see, e.g., Lee et al., Int. J. Pharm. 233:1-18 (2002)). In one
embodiment, a compound described herein is delivered by a topical
ocular delivery method (see, e.g., Curr. Drug Metab. 4:213-22
(2003)). The composition may be in the form of an eye drop, salve,
or ointment or the like, such as, aqueous eye drops, aqueous
ophthalmic suspensions, non-aqueous eye drops, and non-aqueous
ophthalmic suspensions, gels, ophthalmic ointments, etc. For
preparing a gel, for example, carboxyvinyl polymer, methyl
cellulose, sodium alginate, hydroxypropyl cellulose, ethylene
maleic anhydride polymer and the like can be used.
[1077] The dose of the composition comprising at least one of the
compounds described herein may differ, depending upon the patient's
(e.g., human) condition, that is, stage of the disease, general
health status, age, and other factors that a person skilled in the
medical art will use to determine dose. When the composition is
used as eye drops, for example, one to several drops per unit dose,
preferably 1 or 2 drops (about 50 .mu.l per 1 drop), may be applied
about 1 to about 6 times daily.
[1078] Pharmaceutical compositions may be administered in a manner
appropriate to the disease to be treated (or prevented) as
determined by persons skilled in the medical arts. An appropriate
dose and a suitable duration and frequency of administration will
be determined by such factors as the condition of the patient, the
type and severity of the patient's disease, the particular form of
the active ingredient, and the method of administration. In
general, an appropriate dose and treatment regimen provides the
composition(s) in an amount sufficient to provide therapeutic
and/or prophylactic benefit (e.g., an improved clinical outcome,
such as more frequent complete or partial remissions, or longer
disease-free and/or overall survival, or a lessening of symptom
severity). For prophylactic use, a dose should be sufficient to
prevent, delay the onset of, or diminish the severity of a disease
associated with neurodegeneration of retinal neuronal cells and/or
degeneration of other mature retinal cells such as RPE cells.
Optimal doses may generally be determined using experimental models
and/or clinical trials. The optimal dose may depend upon the body
mass, weight, or blood volume of the patient.
[1079] The doses of the compounds described herein can be suitably
selected depending on the clinical status, condition and age of the
subject, dosage form and the like. In the case of eye drops, a
compound described herein can be administered, for example, from
about 0.01 mg, about 0.1 mg, or about 1 mg, to about 25 mg, to
about 50 mg, to about 90 mg per single dose. Eye drops can be
administered one or more times per day, as needed. In the case of
injections, suitable doses can be, for example, about 0.0001 mg,
about 0.001 mg, about 0.01 mg, or about 0.1 mg to about 10 mg, to
about 25 mg, to about 50 mg, or to about 90 mg of the compound
described herein, one to seven times per week. In other
embodiments, about 1.0 to about 30 mg of the compound described
herein can be administered one to seven times per week.
[1080] Oral doses can typically range from 1.0 to 1000 mg, one to
four times, or more, per day. An exemplary dosing range for oral
administration is from 10 to 250 mg one to three times per day. If
the composition is a liquid formulation, the composition comprises
at least 0.1% active compound at particular mass or weight (e.g.,
from 1.0 to 1000 mg) per unit volume of carrier, for example, from
about 2% to about 60%.
[1081] In certain embodiments, at least one compound described
herein may be administered under conditions and at a time that
inhibits or prevents dark adaptation of rod photoreceptor cells. In
certain embodiments, the compound is administered to a subject at
least 30 minutes (half hour), 60 minutes (one hour), 90 minutes
(1.5 hour), or 120 minutes (2 hours) prior to sleeping. In certain
embodiments, the compound may be administered at night before the
subject sleeps. In other embodiments, a light stimulus may be
blocked or removed during the day or under normal light conditions
by placing the subject in an environment in which light is removed,
such as placing the subject in a darkened room or by applying an
eye mask over the eyes of the subject. When the light stimulus is
removed in such a manner or by other means contemplated in the art,
the agent may be administered prior to sleeping.
[1082] The doses of the compounds that may be administered to
prevent or inhibit dark adaptation of a rod photoreceptor cell can
be suitably selected depending on the clinical status, condition
and age of the subject, dosage form and the like. In the case of
eye drops, the compound (or the composition comprising the
compound) can be administered, for example, from about 0.01 mg,
about 0.1 mg, or about 1 mg, to about 25 mg, to about 50 mg, to
about 90 mg per single dose. In the case of injections, suitable
doses can be, for example, about 0.0001 mg, about 0.001 mg, about
0.01 mg, or about 0.1 mg to about 10 mg, to about 25 mg, to about
50 mg, or to about 90 mg of the compound, administered any number
of days between one to seven days per week prior to sleeping or
prior to removing the subject from all light sources. In certain
other embodiments, for administration of the compound by eye drops
or injection, the dose is between 1-10 mg (compound)/kg (body
weight of subject) (i.e., for example, 80-800 mg total per dose for
a subject weighing 80 kg). In other embodiments, about 1.0 to about
30 mg of compound can be administered one to seven times per week.
Oral doses can typically range from about 1.0 to about 1000 mg,
administered any number of days between one to seven days per week.
An exemplary dosing range for oral administration is from about 10
to about 800 mg once per day prior to sleeping. In other
embodiments, the composition may be delivered by intravitreal
administration.
[1083] Also contemplated are compounds of the present disclosure
wherein one or more atoms in the molecule are isotopically
enriched. In one embodiment, the compound is enriched with
deuterium. In another embodiment, the compound is enriched with an
isotope selected from 2H, .sup.11C, .sup.13C, .sup.14C, .sup.15C,
.sup.12N, .sup.13N, .sup.15N, .sup.16N, .sup.16O, .sup.17O,
.sup.14F, .sup.15F, .sup.16F, .sup.17F, .sup.18F, .sup.33S,
.sup.34S, .sup.35S, .sup.36S, .sup.35Cl, .sup.37Cl, .sup.79Br,
.sup.81Br, or .sup.125I. In one embodiment, the enrichment is no
less than 98%. In one embodiment, the enrichment is no less than
95%. In one embodiment, the enrichment is no less than 90%. In one
embodiment, the enrichment is no less than 75%. In one embodiment,
the enrichment is no less than 50%. In one embodiment, the
enrichment is no less than 20%. In one embodiment, the enrichment
is no less than 10%. In one embodiment, the enrichment is no less
than 5%. In one embodiment, the enrichment is no less than 1%.
Ratios of enrichment are determined by mass spectroscopy.
[1084] Isotopically enriched compounds provide improved
pharmaceutical properties compared to the non-enriched compounds.
In many cases this is a result of kinetic isotope effect arising
during ADME processes. In one embodiment, the isotopically enriched
compound of the present disclosure has improved pharmacokinetic
properties compared to the non-isotopically enriched compound of
the present disclosure. In one embodiment, the isotopically
enriched compound of the present disclosure has an increased AUC
compared to the non-isotopically enriched compound of the present
disclosure. In one embodiment, the isotopically enriched compound
of the present disclosure has reduced first-pass effect compared to
the non-isotopically enriched compound of the present disclosure.
In one embodiment, the isotopically enriched compound of the
present disclosure has an increased half-life of elimination
compared to the non-isotopically enriched compound of the present
disclosure. In one embodiment, the isotopically enriched compound
of the present disclosure has improved drug-drug interaction
properties compared to the non-isotopically enriched compound of
the present disclosure. In one embodiment, the isotopically
enriched compound of the present disclosure has different
metabolite profile compared to the non-isotopically enriched
compound of the present disclosure. In one embodiment, the
isotopically enriched compound of the present disclosure has a
reduced rate of oxidation in vivo compared to the non-isotopically
enriched compound of the present disclosure. In one embodiment, the
isotopically enriched compound of the present disclosure has a
reduced cytochrome p450 inhibition propensity compared to the
non-isotopically enriched compound of the present disclosure. In
one embodiment, the isotopically enriched compound of the present
disclosure has a different cytochrome p450 inhibition profile
compared to the non-isotopically enriched compound of the present
disclosure. In one embodiment, the isotopically enriched compound
of the present disclosure has a reduced cytochrome p450 induction
propensity compared to the non-isotopically enriched compound of
the present disclosure.
[1085] Also provided are methods of manufacturing the compounds and
pharmaceutical compositions described herein. A composition
comprising a pharmaceutically acceptable excipient or carrier and
at least one of the compounds described herein may be prepared by
synthesizing the compound according to any one of the methods
described herein or practiced in the art and then formulating the
compound with a pharmaceutically acceptable carrier. Formulation of
the composition will be appropriate and dependent on several
factors, including but not limited to, the delivery route, dose,
and stability of the compound.
[1086] Other embodiments and uses will be apparent to one skilled
in the art in light of the present disclosures. The following
examples are provided merely as illustrative of various embodiments
and shall not be construed to limit the invention in any way.
[1087] While preferred embodiments of the present invention have
been shown and described herein, it will be obvious to those
skilled in the art that such embodiments are provided by way of
example only. Numerous variations, changes, and substitutions will
now occur to those skilled in the art without departing from the
invention. It should be understood that various alternatives to the
embodiments of the invention described herein may be employed in
practicing the invention. It is intended that the following claims
define the scope of the invention and that methods and structures
within the scope of these claims and their equivalents be covered
thereby.
EXAMPLES
[1088] Unless otherwise noted, reagents and solvents were used as
received from commercial suppliers. Anhydrous solvents and
oven-dried glassware were used for synthetic transformations
sensitive to moisture and/or oxygen. Yields were not optimized.
Reaction times are approximate and were not optimized. Flash column
chromatography and thin layer chromatography (TLC) were performed
on silica gel unless otherwise noted. Proton and carbon nuclear
magnetic resonance spectra were obtained with a Varian VnmrJ 400 at
400 MHz for proton and 100 MHz for carbon, or with a Bruker 400 MHz
with Multi Probe/Dual Probe at 400 MHz for proton and 100 MHz for
carbon, as noted. Spectra are given in ppm (.delta.) and coupling
constants, J are reported in Hertz. For proton spectra either
tetramethylsilane was used as an internal standard or the solvent
peak was used as the reference peak. For carbon spectra the solvent
peak was used as the reference. Mass-spectra were recorded using
electrospray ionization (ES+) mode in Agilent LC/MSD SL
mass-spectrometer or (ES+/ES-) mode in Waters Single Quadrupole
Detector. Chiral HPLC analysis was performed using a Chiralpak IA
column (4.6.times.250 mm, 50 on an Agilent HP 1100 system with
diode array detection with heptane-EtOH with 0.1% ethanesulfonic
acid as an eluent.
Analytical HPLC Methods
[1089] Method 1. Column: Phenomenex Gemini (150.times.4.6
mm.times.5.mu.); Flow Rate: 1.0 mL/min; Detection at 220 nm using
DAD; Column temperature 30.degree. C.; Solvent A: 0.05% TFA in
water, Solvent B: 0.05% TFA in acetonitrile; Run Time: 24 min;
Gradient program:
TABLE-US-00002 Time (min) Solvent A (%) Solvent B (%) 0 90 10 15 30
70 17 5 95 20 5 95 20.01 90 10 24 90 10
[1090] Method 2. Column: Phenomenex Gemini (150.times.4.6
mm.times.5.mu.); Flow Rate: 1.0 mL/min; Detection at 220 nm using
DAD; Column temperature 30.degree. C.; Solvent A: 0.05% TFA in
water, Solvent B: 0.05% TFA in acetonitrile; Run Time: 24 min;
Gradient program:
TABLE-US-00003 Time (min) Solvent A (%) Solvent B (%) 0 90 30 6 30
80 9 5 95 12 5 95 13 90 30 16 90 30
[1091] Method 3. Column: Acquity Shield RP-18 (2.1.times.100 mm,
1.7 .mu.m); Flow Rate: 0.3 mL/min; Detection at 214 nm using DAD;
Column temperature 30.degree. C.; Solvent A: 0.1% TFA in water,
Solvent B: acetonitrile; Run Time: 10 min; Gradient program:
TABLE-US-00004 Time (min) Solvent A (%) Solvent B (%) 0.0 90 10 1.0
90 10 2.0 85 15 4.5 55 55 6.0 10 90 8.0 10 90 9.0 90 10 10.0 90
10
[1092] Method 4. Column: Acquity Shield RP-18 (2.1.times.100 mm,
1.7 .mu.m); Flow Rate: 0.3 mL/min; Detection at 214 nm using DAD;
Column temperature 30.degree. C.; Solvent A: 0.1% TFA in water,
Solvent B: MeOH; Run Time: 10 min; Gradient program was same as for
Method 3.
[1093] Method 5. Column: Waters Acquity C-8 (2.1.times.100 mm, 1.7
.mu.m); Flow Rate: 0.3 mL/min; Detection at 214 nm using DAD;
Column temperature 30.degree. C.; Solvent A: 5 mM KH.sub.2PO.sub.4,
Solvent B: acetonitrile; Run Time: 10 min; Gradient program was
same as for Method 3.
[1094] Method 6. Column: Acquity BEH C-18 (2.1.times.100 mm, 1.7
.mu.m); Flow Rate: 0.3 mL/min; Detection at 247 nm using DAD;
Column temperature 30.degree. C.; Solvent A: 5 mM Ammonium Acetate
in water, Solvent B: acetonitrile; Run Time: 10 min; Gradient
program was same as for Method 3.
Example 1
Preparation of N-(3-(2-aminoethoxy)phenyl)pentane-2-sulfonamide
##STR00289##
[1096] N-(3-(2-Aminoethoxy)phenyl)pentane-2-sulfonamide was
prepared following the method shown in Scheme 1.
##STR00290##
[1097] Step 1: A mixture of 1-aminophenol (1) (207 mg, 1.9 mmol),
2-(tert-butoxycarbonylamino)ethyl 4-methylbenzenesulfonate (2) (500
mg, 1.9 mmol), and cesium carbonate (770 mg, 2.2 mmol) in DMF (6
ml) was stirred at room temperature under argon for 15 hours. The
mixture was concentrated under reduced pressure. The residue was
partitioned between EtOAc and water. The organic layer was washed
with brine, dried over Na.sub.2SO.sub.4 and concentrated under
reduced pressure. Purification by flash chromatography (40 to 60%
EtOAc-hexanes gradient) gave tert-butyl
2-(3-aminophenoxy)ethylcarbamate (3) as colorless oil. Yield (220
mg, 58%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 6.92 (t,
J=5.2 Hz, 1H), 6.85 (t, J=8.0 Hz, 1H), 6.08-6.12 (m, 2H), 6.02-6.04
(m, 1H), 4.99 (bs, 2H), 3.79 (t, J=6.0 Hz, 2H), 3.21 (q, J=6.0 Hz,
2H), 1.36 (s, 9H).
[1098] Step 2: A mixture of tert-butyl
2-(3-aminophenoxy)ethylcarbamate (3) (210 mg, 1.1 mmol),
2-pentylsulfonyl chloride (4) (0.17 ml, 1.1 mmol) and DMAP (20 mg)
in pyridine (5 ml) was stirred at room temperature under argon for
15 hours. The solvent was evaporated under reduced pressure. The
residue was partitioned between EtOAc and 0.5 N HCl aq. The organic
layer was washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure. Purification by flash
chromatography (40 to 60% EtOAc-hexanes gradient) gave tert-butyl
2-(3-(1-methylbutylsulfonamido)phenoxy)ethylcarbamate (5) as light
yellow oil. Yield (160 mg, 46%). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.20 (t, J=8.0 Hz, 1H), 6.75-6.82 (m, 1H), 6.73-6.75 (m,
1H), 6.64-6.67 (m, 1H), 6.37 (bs, 1H), 4.96 (bs, 1H), 3.99 (t,
J=6.0 Hz, 2H), 3.51 (q, J=6.0 Hz, 2H), 3.11-3.18 (m, 1H), 1.91-2.0
(m, 1H), 1.53-1.60 (m, 2H), 1.44 (s, 9H), 1.23-1.37 (m, 4H),
0.88-0.91 (m, 3H).
[1099] Step 3: A mixture of tert-butyl
2-(3-(1-methylbutylsulfonamido)phenoxy)ethylcarbamate (5) (160 mg,
0.48 mmol) and HCl-EtOH (6.95 M, 3.0 ml) in ethyl acetate (5 ml)
was stirred at room temperature for 15 hours. The solvent was
evaporated under reduced pressure. A mixture of EtOAc-hexane (30%,
5 ml) was added and the mixture was sonicated. The solid was
collected by filtration and dried to give Example 1 as a white
solid. Yield (80 mg, 69%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.81 (s, 1H), 8.07 (bs, 3H), 7.21 (t, J=2.4 Hz, 1H), 6.81
(dd, J=8.4, 2.4 Hz, 1H), 6.56 (dd, J=8.4, 2.4 Hz, 1H), 4.09 (t,
J=6.8 Hz, 2H), 3.18 (q, J=6.0 Hz, 2H), 3.01-3.09 (m, 1H), 1.74-1.83
(m, 1H), 1.33-1.46 (m, 2H), 1.18-1.28 (m, 4H), 0.79 (t, J=7.2 Hz,
3H).
Example 2
Preparation of N-(3-(2-aminoethoxy)phenyl)butane-2-sulfonamide
##STR00291##
[1101] N-(3-(2-Aminoethoxy)phenyl)butane-2-sulfonamide was prepared
following the method described in Example 1.
[1102] Step 1: Sulfonation of tert-butyl
2-(3-aminophenoxy)ethylcarbamate (3) using butane-2-sulfonyl
chloride following the method described in Example 1 gave
tert-butyl 2-(3-(1-methylpropylsulfonamido)phenoxy)ethylcarbamate
(6) as a light yellow oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.72 (s, 1H), 7.17 (t, J=8.0 Hz, 1H), 6.97 (t, J=6.0 Hz,
1H), 6.76-6.78 (m, 2H), 6.59-6.62 (m, 1H), 3.87 (t, J=5.6 Hz, 2H),
3.24 (q, J=6.4 Hz, 2H), 2.93-3.02 (m, 1H), 1.80-1.91 (m, 1H),
1.40-1.48 (m, 1H), 1.35 (s, 9H), 1.19 (d, J=6.8 Hz, 3H), 0.88 (t,
J=7.2 Hz, 3H).
[1103] Step 2: Deprotection of tert-butyl
2-(3-(1-methylpropylsulfonamido)phenoxy)ethylcarbamate (6)
following the method described in Example 1 gave Example 2 as a
colorless oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.16 (t,
J=8.0 Hz, 1H), 6.75-6.77 (m, 2H), 6.59-6.62 (m, 1H), 3.83 (t, J=5.2
Hz, 2H), 2.93-3.02 (m, 1H), 2.83 (t, J=5.6 Hz, 2H), 1.80-1.91 (m,
1H), 1.38-1.48 (m, 1H), 1.19 (d, J=6.8 Hz, 3H), 0.88 (t, J=7.2 Hz,
3H).
Example 3
Preparation of N-(3-(2-aminoethoxy)phenyl)propane-2-sulfonamide
##STR00292##
[1105] N-(3-(2-Aminoethoxy)phenyl)propane-2-sulfonamide was
prepared following the method described in Example 1.
[1106] Step 1: Sulfonation of tert-butyl
2-(3-aminophenoxy)ethylcarbamate (3) using propane-2-sulfonyl
chloride following the method described in Example 1 gave
tert-butyl 2-(3-(1-methylethylsulfonamido)phenoxy)ethylcarbamate
(7) as a light yellow oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.70 (s, 1H), 7.17 (t, J=8.0 Hz, 1H), 6.97 (t, J=6.0 Hz,
1H), 6.76-6.79 (m, 2H), 6.60-6.62 (m, 1H), 3.16-3.30 (m, 3H), 1.35
(s, 9H), 1.19 (d, J=6.8 Hz, 6H).
[1107] Step 2: Deprotection of tert-butyl
2-(3-(1-methylethylsulfonamido)phenoxy)ethylcarbamate (7) following
the method described in Example 1 gave Example 3 as a white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.80 (s, 1H), 8.07 (bs,
3H), 7.21 (t, J=8.0 Hz, 1H), 6.86 (t, J=2.0 Hz, 1H), 6.81 (dd,
J=8.0, 2.0 Hz, 1H), 6.66 (dd, J=8.0, 2.0 Hz, 1H), 4.10 (t, J=5.2
Hz, 2H), 3.15-3.24 (m, 3H), 1.20 (d, J=6.8 Hz, 6H).
Example 4
Preparation of
N-(3-(2-aminoethoxy)phenyl)cyclohexanesulfonamide
##STR00293##
[1109] N-(3-(2-Aminoethoxy)phenyl)cyclohexanesulfonamide was
prepared following the method described in Example 1.
[1110] Step 1: Sulfonation of tert-butyl
2-(3-aminophenoxy)ethylcarbamate (3) using cyclohexanesulfonyl
chloride (8) following the method described in Example 1 gave
tert-butyl 2-(3-(cyclohexanesulfonamido)phenoxy)ethylcarbamate (9)
as a light yellow oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.70 (s, 1H), 7.16 (t, J=8.0 Hz, 1H), 6.97 (t, J=6.0 Hz, 1H),
6.75-6.78 (m, 2H), 6.59-6.62 (m, 1H), 3.87 (t, J=5.6 Hz, 2H), 3.24
(q, J=6.0 Hz, 2H), 2.89-2.98 (m, 1H), 1.92-2.01 (m, 2H), 1.68-1.76
(m, 2H), 1.52-1.57 (m, 1H), 1.31-42 (m, 11H), 1.05-1.22 (m,
2H).
[1111] Step 2: Deprotection of tert-butyl
2-(3-(cyclohexanesulfonamido)phenoxy)ethylcarbamate (9) following
the method described in Example 1 gave Example 4 as a white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.79 (s, 1H), 8.07 (bs,
3H), 7.20 (t, J=8.0 Hz, 1H), 6.79-6.85 (m, 2H), 6.64 (d, J=7.6 Hz,
1H), 4.07 (t, J=4.8 Hz, 2H), 3.12-3.18 (m, 2H), 2.93 (t, J=7.6 Hz,
1H), 2.89-2.98 (m, 1H), 1.92-2.01 (m, 2H), 1.68-1.76 (m, 2H),
1.52-1.57 (m, 1H), 1.32-42 (m, 2H), 1.00-1.21 (m, 2H).
Example 5
Preparation of
N-(3-(3-amino-1-hydroxypropyl)phenyl)cyclohexanesulfonamide
##STR00294##
[1113] N-(3-(3-Amino-1-hydroxypropyl)phenyl)cyclohexanesulfonamide
was prepared following the method shown in Scheme 2.
##STR00295##
[1114] Step 1: To a solution of CH.sub.3CN (0.7 ml, 16 mmol) in THF
(10 ml) was added LDA (8 ml, 2M in THF, 16 mmol) at -78.degree. C.
and the mixture was stirred at this temperature for 10 min. A
chilled (-78.degree. C.) solution of nitrobenzaldehye (10) (2.0 g,
13 mmol) in THF (15 ml) was added slowly. The resulting mixture was
stirred at -78.degree. C. for 15 mins. The reaction was quenched by
the addition of sat. NH.sub.4Cl aq (10 ml) and the mixture allowed
to warm to room temperature. The organic layer was collected and
the aqueous layer was extracted with EtOAc. The combined organic
layers were dried over Na.sub.2SO.sub.4 and concentrated under
reduced pressure. Purification by flash chromatography (30 to 65%
EtOAc-hexanes gradient) gave
3-hydroxy-3-(3-nitrophenyl)propanenitrile (11) as colorless oil,
Yield (1.9 g, 77%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.30
(t, J=3.2 Hz, 1H), 8.21-8.24 (m, 1H), 7.76-7.80 (m, 1H), 7.71 (t,
J=8.0 Hz, 1H), 5.20 (t, J=2.4 Hz, 1H), 2.22 (d, J=6.0 Hz, 2H), 1.24
(s, 1H).
[1115] Step 2: A mixture of
3-hydroxy-3-(3-nitrophenyl)propanenitrile (11) (400 mg, 2.1 mmol)
and Pd/C (20 mg, 10%) in EtOAc (15 ml) was degassed vacuum/hydrogen
and then stirred at room temperature under H.sub.2 (balloon) for 15
hours. The mixture was filtered to remove the Pd/C and then
concentrated under reduced pressure to give
3-(3-aminophenyl)-3-hydroxypropanenitrile (12) as while solid.
Yield (390 mg, 99%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.15
(t, J=8.0 Hz, 1H), 6.70-6.74 (m, 2H), 6.63-6.65 (m, 1H), 4.92 (t,
J=6.0 Hz, 1H), 2.72 (d, J=6.0 Hz, 2H).
[1116] Step 3: Sulfonation of
3-(3-aminophenyl)-3-hydroxypropanenitrile (12) following the method
described in Example 1 gave
N-(3-(2-cyano-1-hydroxyethyl)phenyl)cyclohexanesulfonamide (13) as
a light yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.74
(s, 1H), 7.23-7.29 (m, 2H), 7.04-7.11 (m, 2H), 5.94 (d, J=2.8 Hz,
1H), 4.83 (q, J=5.2 Hz, 1H), 2.92-3.01 (m, 1H), 2.72-2.86 (m, 2H),
1.91-2.02 (m, 2H), 1.63-1.73 (m, 2H), 1.50-1.57 (m, 1H), 1.31-1.42
(m, 2H), 1.02-1.22 (m, 3H).
[1117] Step 4: BH.sub.3-Me.sub.2S (1.2 ml, 12.7 mmol) was added
under argon to a solution of
N-(3-(2-cyano-1-hydroxyethyl)phenyl)cyclohexanesulfonamide (1.2 g,
3.9 mmol) in anhydrous THF. The reaction mixture was stirred at
60.degree. C. for 18 hrs. The reaction was quenched by the addition
of 2N HCl to pH 0 and stirred at room temperature for 24 hrs. The
pH was then adjusted to 10 by adding 50% aq. NaOH. MTBE (40 ml) was
added to the mixture and stirred. Organic layer was dried over
anhydrous Na.sub.2SO.sub.4 and concentrated under reduced pressure
to give Example 5 as a white solid. Yield (0.60 g, 49%); .sup.1H MR
(400 MHz, CDCl.sub.3) .delta. 7.20-7.28 (m, 2H), 7.11-7.13 (m, 2H),
4.94 (dd, J=8.8, 2.4 Hz, 1H), 3.06-3.15 (m, 1H), 2.94-3.04 (m, 2H),
2.10-2.18 (m, 2H), 1.78-1.90 (m, 3H), 1.50-1.76 (m, 4H), 1.10-1.26
(m, 3H).
Example 6
Preparation of
N-(3-(3-aminopropyl)phenyl)cyclohexanesulfonamide
##STR00296##
[1119] N-(3-(3-Aminopropyl)phenyl)cyclohexanesulfonamide was
prepared following the method shown in Scheme 3.
##STR00297##
[1120] Step 1: To an oven dried, argon filed, flask was added
1-bromo-3-nitrobenzene (14) (3.09 g, 15.3 mmol), tert-butyl
prop-2-ynylcarbamate (15) (2.8 g, 18.0 mmol), diisopropylamine
(92.5 ml, 17.8 mmol), CuI (0.054 g, 0.18 mmol),
PdCl.sub.2(PPh.sub.3).sub.2 (0.42 g, 0.6 mmol) and dioxane (17 ml).
The resulting mixture was purged with argon three times and then a
solution of t-Bu.sub.3P-dioxane solution (0.9 ml, 0.9 mmol) was
added. The mixture was heated at 45.degree. C. for 15 h, cooled to
room temperature, diluted with ethyl acetate, filtered through
celite and the filtrate was concentrated under reduced pressure.
Purification by flash chromatography (10 to 50% EtOAc-hexanes
gradient) gave tert-butyl 3-(3-nitrophenyl)prop-2-ynylcarbamate
(16) as colorless oil. Yield (4.98 g, 100%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.25 (t, J=1.6 Hz, 1H), 8.15 (ddd, J=8.4, 2.4,
1.2 Hz, 1H), 7.69 (dt, J=7.6, 0.8 Hz, 1H), 4.78 (br s, 1H), 4.16
(d, J=5.6 Hz, 2H), 1.47 (s, 9H).
[1121] Step 2: Hydrogenation of tert-butyl
3-(3-nitrophenyl)prop-2-ynylcarbamate (16) following method
described in Example 5 gave tert-butyl
3-(3-aminophenyl)propylcarbamate (17) as a light yellow oil. Yield
(2.57 g, 78%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.04-7.08
(m, 1H), 6.57-6.59 (m, 1H), 7.51-7.29 (m, 2H), 4.50 (br s, 1H),
3.13 (q, J=6.8 Hz, 2H), 2.54 (t, J=7.6 Hz, 2H), 1.73-1.81 (m, 2H),
1.43 (s, 9H).
[1122] Step 3: Sulfonation of tert-butyl
3-(3-aminophenyl)propylcarbamate (17) following the method
described in Example 1 except pyridine and DMAP were used instead
of TEA and DCM gave tert-butyl
3-(3-(cyclohexanesulfonamido)phenyl)propylcarbamate (18) as a
colorless oil. Yield (0.2 g, 32%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.47 (s, 1H), 7.18 (t, J=8.0 Hz, 1H), 7.05-7.09
(m, 2H), 6.90 (d, J=7.6 Hz, 1H), 4.64 (br s, 1H), 3.05-3.16 (m,
2H), 2.93-3.01 (m, 1H), 2.58 (t, J=7.6 Hz, 2H), 2.12-2.15 (m, 2H),
1.72-1.83 (m, 4H), 1.48-1.64 (m, 3H), 1.42 (s, 9H), 1.10-1.24 (m,
3H).
[1123] Step 4: Deprotection of tert-butyl
3-(3-(cyclohexanesulfonamido)phenyl)propylcarbamate (18) following
method described in Example 1 except that the hydrochloride salt
was converted to the free amine by washing the organic solution
with aqueous NaHCO.sub.3 to give Example 6 as a colorless oil.
Yield (0.071 g, 43%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.13-7.17 (m, 1H), 6.95-7.01 (m, 2H), 6.88-6.90 (m, 1H), 2.89-2.96
(m, 1H), 2.57 (t, J=7.6 Hz, 2H), 2.07-2.10 (m, 2H), 1.68-1.84 (m,
4H), 1.46-1.61 (m, 3H), 1.08-1.22 (m, 3H).
Example 7
Preparation of 3-(3-aminopropyl)-N-(cyclohexylmethyl)aniline
##STR00298##
[1125] 3-(3-Aminopropyl)-N-(cyclohexylmethyl)aniline hydrochloride
was prepared following the method shown in Scheme 4.
##STR00299##
[1126] Step 1: A mixture of tert-butyl
3-(3-(cyclohexylmethylamino)phenyl)propylcarbamate (17) (0.31 g,
1.22 mmol), cyclohexanecarbonitrile (19) (0.73 ml, 6.1 mmol),
ammonium acetate (0.1 g, 1.29 mmol) in MeOH (20 ml) was purged with
argon. Pd/C (10%, 0.04 g) was added and the atmosphere exchange
with hydrogen. The mixture was stirred under H.sub.2 (balloon) for
18 h at room temperature. The Pd/C was removed by filtration
through celite, and the filtrate was concentrated under reduced
pressure. Purification by flash chromatography (0 to 50%
EtOAc-hexanes gradient) gave tert-butyl
3-(3-(cyclohexylmethylamino)phenyl)propylcarbamate (20) as a
colorless oil. Yield (0.33 g, 78%); .sup.1Yield (0.2 g, 32%);
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.06 (t, J=8.0 Hz, 1H),
6.48-6.49 (m, 1H), 6.39-6.44 (m, 2H), 4.50 (br s, 1H), 3.14 (q,
J=6.8 Hz, 2H), 2.92 (d, J=6.8 Hz, 2H), 2.54 (t, J=7.6 Hz, 2H),
1.65-1.83 (m, 7H), 1.50-1.61 (m, 1H), 1.43 (s, 9H), 1.11-1.29 (m,
3H), 0.92-1.02 (m, 2H).
[1127] Step 2: Deprotection of tert-butyl
3-(3-(cyclohexylmethylamino)phenyl)propylcarbamate (20) following
method described in Example 1 gave Example 7 hydrochloride salt as
a white solid. Yield (0.29 g, 98%); .sup.1H NMR (400 MHz, MeOD)
.delta. 7.51 (t, J=8.0 Hz, 1H), 7.38-7.43 (m, 2H), 7.32-7.37 (m,
1H), 3.24 (d, J=6.8 Hz, 2H), 2.97 (t, J=7.6 Hz, 2H), 2.78 (t, J=7.6
Hz, 2H), 1.95-2.03 (m, 2H), 1.68-1.88 (m, 6H), 1.22-1.36 (m, 3H),
1.06-1.18 (m, 2H).
Example 8
Preparation of N-(3-(3-aminopropyl)phenyl)cyclohexanecarboxamide
hydrochloride
##STR00300##
[1129] N-(3-(3-Aminopropyl)phenyl)cyclohexanecarboxamide
hydrochloride was prepared following the method shown in Scheme
5.
##STR00301##
[1130] Step 1: To a mixture of cyclohexanecarboxylic acid (21)
(0.23 ml, 1.79 mmol), TBTU (0.56 g, 1.74 mmol) and iPr.sub.2EtN
(0.33 ml, 1.89 mmol) in DMF (20 ml) was added tert-butyl
3-(3-(cyclohexylmethylamino)phenyl)propylcarbamate (17) (0.40 g,
1.59 mmol) in DMF (5 ml). The mixture was stirred at room
temperature for 18 h and then diluted with water. The solution was
extracted with ethyl acetate and the combined extracts were washed
with water, aqueous NaHCO.sub.3 and brine, dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure.
Purification by flash chromatography (5 to 50% EtOAc-hexanes
gradient) gave tert-butyl
3-(3-(cyclohexanecarboxamido)phenyl)propylcarbamate (22) as a
colorless oil. Yield (0.455 g, 78%); Yield (0.2 g, 32%); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.43 (br s, 1H), 7.26-7.28 (m,
2H), 7.20 (t, J=7.6 Hz, 1H), 7.08 (br s, 1H), 6.89-6.91 (m, 1H),
3.12 (t, J=6.0 Hz, 2H), 2.61 (t, J=8.4 Hz, 2H), 2.17-2.23 (m, 1H),
1.93-1.96 (m, 2H), 1.74-1.86 (m, 3H), 1.65-1.72 (m, 2H), 1.48-1.59
(m, 1H), 1.31-1.42 (m, 2H), 1.43 (s, 9H), 1.22-1.38 (m, 4H).
[1131] Step 2: Deprotection of tert-butyl
3-(3-(cyclohexanecarboxamido)phenyl)propylcarbamate (22) following
method described in Example 1 gave Example 8 hydrochloride salt as
a white solid. Yield (0.31 g, 92%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.76 (s, 1H), 7.81 (br s, 3H), 7.56 (s, 1H),
7.20-7.32 (m, 1H), 7.17 (t, J=8.0 Hz, 1H), 6.83 (d, J=6.8 Hz, 1H),
2.70-2.81 (m, 2H), 2.55 (t, J=7.6 Hz, 2H), 2.26-2.34 (m, 1H),
1.61-1.83 (m, 7H), 1.13-1.41 (m, 5H).
Example 9
Preparation of 3-(3-(3-aminopropyl)phenyl)-1,1-dipropylurea
##STR00302##
[1133] 3-(3-(3-Aminopropyl)phenyl)-1,1-dipropylurea was prepared
following the method shown in Scheme 6.
##STR00303##
[1134] Step 1: A mixture of 1-bromo-3-isocyanatobenzene (22) (1.044
g, 5.27 mmol) and dipropylamine (23) (0.75 mL, 5.80 mmol) in
anhydrous THF was stirred at room temperature for 1 hr. The mixture
was concentrated under reduced pressure. Crystallization from
hexanes gave 3-(3-bromophenyl)-1,1-dipropylurea (24) as a white
solid. Yield (1.512 g, 96%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.25 (br.s, 1H), 7.76 (t, J=2.0 Hz, 1H), 7.45 (ddd, J=1.2,
2.2, 8.2 Hz, 1H), 7.14 (t, J=8.0 Hz, 1H), 7.06 (ddd, J=1.0, 2.0,
7.8 Hz, 1H), 3.21 (t, J=7.4 Hz, 4H), 1.47 (sextet, J=7.4 Hz, 4H),
0.82 (t, J=7.2 Hz, 6H).
[1135] Step 2: A solution of 3-(3-bromophenyl)-1,1-dipropylurea
(24) (0.507 g, 1.70 mmol), tert-butyl prop-2-ynylcarbamate (15)
(0.323 g, 2.12 mmol), tri-o-tolylphosphine (0.0342 g, 0.112 mmol)
and Et.sub.3N (3.0 mL) in DMF was degassed by bubbling argon for 10
min, and applying vacuum/argon 3.times..
PdCl.sub.2(Ph.sub.3P).sub.2 (0.0434 g, 0.062 mmol) followed by CuI
(0.0263 g, 0.138 mmol) were added and the mixture was degassed by
applying vacuum/argon 3.times.. The reaction mixture was stirred
under argon at 70.degree. C. for 22 hrs. The reaction mixture was
concentrated under reduced pressure. Purification by flash
chromatography (10% to 50% EtOAc-hexanes gradient) gave tert-butyl
3-(3-(3,3-dipropylureido)phenyl)prop-2-ynylcarbamate (25) as a pale
yellow solid. Yield (0.174 g, 28%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.15 (br.s, 1H), 7.56 (t, J=1.8 Hz, 1H),
7.42-7.46 (m, 1H), 7.31 (br.t, 1H), 7.17 (t, J=7.8 Hz, 1H),
6.89-6.93 (m, 1H), 3.93 (d, J=5.7 Hz, 2H), 3.21 (t, J=7.6 Hz, 4H),
1.47 (sextet, J=7.4 Hz, 4H), 1.32-1.38 (m, 9H), 0.82 (t, J=7.4 Hz,
6H).
[1136] Step 3: A solution of tert-butyl
3-(3-(3,3-dipropylureido)phenyl)prop-2-ynylcarbamate (25) (0.17 g,
0.455 mmol) in EtOH (10 mL) was degassed with vacuum/Ar. Pd/C (10%,
0.0293 g) was added and the atmosphere was purged with H.sub.2. The
mixture was stirred under H.sub.2-filled balloon at room
temperature for 5 hrs. The reaction mixture was filtered through
Celite and concentrated under reduced pressure. The residue was
crystallized from EtOAc/hexanes to give tert-butyl
3-(3-(3,3-dipropylureido)phenyl)propylcarbamate (26) as a pale
yellow solid. Yield (0.0946 g, 55%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.00 (s, 1H), 7.22-7.26 (m, 2H), 7.07 (t,
J=7.6 Hz, 1H), 6.82 (br.t, J=5.1 Hz, 1H), 6.70-6.74 (m, 1H), 3.21
(t, J=7.4 Hz, 4H), 2.90 (q, J=6.1 Hz, 2H), 2.45 (t, J=8.0 Hz, 2H),
1.61 (m, 2H), 1.48 (sextet, J=7.4 Hz, 4H), 1.35 (s, 9H), 0.82 (t,
J=7.4 Hz, 6H).
[1137] Step 4. A mixture of tert-butyl
3-(3-(3,3-dipropylureido)phenyl)propylcarbamate (26) (0.094 g,
0.249 mmol) and HCl/EtOAc (3N, 4.5 mL) in EtOAc was stirred at room
temperature for 1 h. The reaction mixture was concentrated under
reduced pressure and the residue was partitioned between aq.
NaHCO.sub.3 and MTBE. The organic layer was concentrated under
reduced pressure. Purification by flash chromatography (20% 7N
NH.sub.3/MeOH/EtOAc) gave Example 9 as a colorless oil. Yield
(0.0154 g, 22%); .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
7.20-7.22 (m, 1H), 7.12-7.18 (m, 2H), 6.84-6.89 (m, 1H), 2.58-2.69
(m, 4H), 1.78 (p, J=7.6 Hz, 2H), 1.62 (sextet, J=7.6 Hz, 4H), 0.93
(t, J=7.4 Hz, 6H); ESI MS m/z 278.60 [M+H].sup.+.
Example 10
Preparation of 1-(3-(2-aminoethoxy)phenyl)-3-cyclohexylthiourea
##STR00304##
[1139] 1-(3-(2-Aminoethoxy)phenyl)-3-cyclohexylthiourea was
prepared following the method shown in Scheme 7.
##STR00305##
[1140] Step 1: A mixture of isothiocyanatocyclohexane (27) (0.16
mL, 1.17 mmol), tert-butyl 2-(3-aminophenoxy)ethylcarbamate (3)
(0.282 g, 1.12 mmol), DMAP (0.024 g, 0.196 mmol) and Et.sub.3N (0.3
mL, 2.15 mmol) in anhydrous THF was stirred under argon at
50.degree. C. for 24 h. The mixture was concentrated under reduced
pressure. Purification by flash chromatography (30% to 60%
EtOAc-hexanes gradient) gave tert-butyl
2-(3-(3-cyclohexylthioureido)phenoxy)ethylcarbamate (28) as a white
solid. Yield (0.1917 g, 44%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.29 (s, 1H), 7.59 (br.d, J=7.6 Hz, 1H), 7.15 (t, J=8.2 Hz,
1H), 6.97 (br.t, J=5.5 Hz, 1H), 6.85-6.89 (m, 1H), 6.58-6.63 (m,
1H), 4.06 (br.s, 1H), 3.88 (t, J=5.9 Hz, 2H), 3.25 (q, J=5.7 Hz,
2H), 1.82-1.90 (m, 2H), 1.60-1.70 (m, 2H), 1.47-1.57 (m, 1H), 1.36
(s, 9H), 1.07-1.34 (m, 6H).
[1141] Step 2: A mixture of tert-butyl
2-(3-(3-cyclohexylthioureido)phenoxy)ethylcarbamate (28) (0.19 g,
0.483 mmol) and HCl/EtOAc (3N, 5 mL) in EtOAc was stirred at room
temperature for 24 h. A precipitate formed which was collected by
filtration. The solid was dissolved in NH.sub.3/MeOH (7N) and the
resulting solution was concentrated under reduced pressure.
Purification by flash chromatography (5% 7N
NH.sub.3/MeOH/CH.sub.2Cl.sub.2) gave Example 10 as a white solid.
Yield (0.101 g, 71%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.30 (s, 1H), 7.59 (br.d, J=7.6 Hz, 1H), 7.18-7.22 (m, 1H), 7.15
(t, J=8.2 Hz, 1H), 6.85-6.89 (m, 1H), 6.58-6.63 (m, 1H), 4.06
(br.s, 1H), 3.84 (t, J=5.9 Hz, 2H), 2.83 (t, J=5.7 Hz, 2H),
1.82-1.90 (m, 2H), 1.60-1.70 (m, 2H), 1.44-1.59 (m, 3H), 1.07-1.24
(m, 5H).
Example 11
Preparation of
3-amino-1-(3-(cyclohexylmethylamino)phenyl)propan-1-ol
##STR00306##
[1143] 3-Amino-1-(3-(cyclohexylmethylamino)phenyl)propan-1-ol was
prepared following the method shown in Scheme 8.
##STR00307##
[1144] Step 1: A solution of 11 (0.8 g, 4.2 mmol) and
cyclohexanecarbaldehyde (29) (0.5 ml, 4.2 mmol) in EtOAc was
degassed and saturated with argon. To this solution was added 10%
Pd/C (50 mg). The resulting mixture was stirred under H.sub.2 at 1
atm for 18 hrs, filtered through Celite, concentrated under reduced
pressure. Purification by flash chromatography (40 to 50%
EtOAc-hexanes gradient) gave aniline 30 as a light yellow oil which
was used in the next step without further purification. Yield (0.9
g, 70%).
[1145] Step 2: Reduction of hydroxynitrile 30 was done following
the method described in Example 5 with the following exception.
After the reaction was completed, it was cooled to room
temperature, the excess of borane was quenched by careful addition
of MeOH, followed by addition of HCl-MeOH (1.25 M, 10 ml), stirring
at 60.degree. C. for 3 hr. Concentration under reduced pressure
gave amine 31 hydrochloride which was used in next step without
further purification.
[1146] Step 3: A solution of Boc.sub.2O (0.6 g, 2.73 mmol) in
CH.sub.2Cl.sub.2 was added dropwise to a suspension of amine 31
(0.68 g. 2.6 mmol) and TEA (1.0 ml, 5.2 mmol) in dichloromethane at
room temperature. The reaction mixture was stirred at room
temperature for 2 hr, washed with HCl--NH.sub.4Cl.aq (0.5 M, 50
ml), dried with Na.sub.2SO.sub.4 and concentrated under reduced
pressure. Purification by flash chromatography (50% to 60%
EtOAc-hexanes gradient) gave carbamate 32 as an off-white solid.
Yield (0.8 g, 88%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
6.94 (t, J=7.6 Hz, 1H), 6.69 (t, J=4.8 Hz, 1H), 6.49 (s, 1H), 6.40
(d, J=7.6 Hz, 1H), 6.36 (dd, J=8.0, 1.6 Hz, 1H), 5.47 (bs, 1H),
4.97 (bs, 1H), 4.33-4.38 (m, 1H), 2.88-2.98 (m, 2H), 2.79 (t, J=6.0
Hz, 2H), 1.73-1.80 (m, 2H), 1.56-1.70 (m, 5H), 1.44-1.56 (m, 1H),
1.34 (s, 9H), 1.15-1.22 (m, 3H), 0.93-0.98 (m, 2H).
[1147] Step 4: Deprotection of carbamate 32 following method
described in Example 1 gave Example 11 hydrochloride as a white
solid. Yield (0.14 g, 92%); .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.51-7.58 (m, 3H), 7.38-7.41 (m, 1H), 4.92 (dd, J=8.4, 3.6
Hz, 1H), 3.26 (d, J=6.8 Hz, 2H), 3.03-3.16 (m, 2H), 1.63-2.01 (m,
8H), 1.04-1.37 (m, 5H).
Example 12
Preparation of
3-amino-1-(3-(cyclohexylmethylamino)phenyl)propan-1-one
##STR00308##
[1149] 3-Amino-1-(3-(cyclohexylmethylamino)phenyl)propan-1-one was
prepared following the method shown in Scheme 9.
##STR00309##
[1150] Step 1: A mixture of alcohol 32 (0.54 g. 1.32 mmol) and
MnO.sub.2 (0.35 g. 3.96 mmol) in DCM was stirred at room
temperature for 18 hrs. The reaction mixture was filtered through
Celite and concentrated under reduced pressure. Purification by
flash chromatography (30% to 60% EtOAc-hexanes gradient) gave
ketone 33 as a light yellow oil. Yield (0.27 g, 57%); .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.15 (t, J=8.0 Hz, 1H), 7.02-7.07 (m,
2H), 6.73-6.79 (m, 2H), 5.87 (t, J=5.6 Hz, 1H), 3.21 (q, J=6.0 Hz,
2H), 3.03 (t, J=6.8 Hz, 2H), 2.85 (t, J=6.0 Hz, 2H), 1.72-1.81 (m,
2H), 1.46-1.71 (m, 4H), 1.34 (s, 9H), 1.10-1.20 (m, 3H), 0.85-0.97
(m, 2H).
[1151] Step 2: Deprotection of carbamate 33 following method
described in Example 1 gave Example 12 hydrochloride as a white
solid. Yield (0.19 g, 94%); .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.01-8.06 (m, 2H), 7.64-7.72 (m, 2H), 3.50 (t, J=6.0 Hz,
2H), 3.35 (t, J=5.6 Hz, 2H), 3.26 (d, J=6.8 Hz, 2H), 1.67-1.90 (m,
6H), 1.20-1.36 (m, 3H), 1.05-1.16 (m, 2H).
Example 13
Preparation of 3-amino-1-(3-(pentylamino)phenyl)propan-1-ol
##STR00310##
[1153] 3-Amino-1-(3-(pentylamino)phenyl)propan-1-ol was prepared
following the method used in Example 11.
[1154] Step 1: Hydrogenation of nitrobenzene 11 (0.8 g, 4.2 mmol)
and pentanal (0.45 ml, 4.2 mmol) following method described in
Example 11 gave 3-hydroxy-3-(3-(pentylamino)phenyl)propanenitrile
as a light yellow oil. Yield (0.90 g, 77%).
[1155] Step 2: Reduction of
3-hydroxy-3-(3-(pentylamino)phenyl)propanenitrile (0.35 g, 1.51
mmol) following method described in Example 11 gave
3-amino-1-(3-(pentylamino)phenyl)propan-1-ol that was used in next
reaction without further purification. Yield (0.41 g, quant.).
[1156] Step 3: Protection of
3-amino-1-(3-(pentylamino)phenyl)propan-1-ol (0.41 g, 1.51 mmol)
following method described in Example 11 gave tert-butyl
3-hydroxy-3-(3-(pentylamino)phenyl)propylcarbamate as a colorless
oil. Yield (0.4 g, 79%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 6.94 (t, J=7.6 Hz, 1H), 6.69 (t, J=4.8 Hz, 1H), 6.49 (s,
1H), 6.41 (d, J=7.6 Hz, 1H), 6.36 (dd, J=8.0, 1.2 Hz, 1H), 5.41 (t,
J=6.4 Hz, 1H), 4.97 (d, J=4.4 Hz, 1H), 4.37 (q, J=4.4 Hz, 1H),
2.90-2.98 (m, 2H), 1.61 (q, J=6.8 Hz, 2H), 1.46-1.56 (m, 2H),
1.26-1.36 (m, 15H), 0.93-0.98 (m, 3H).
[1157] Step 4: Deprotection of tert-butyl
3-hydroxy-3-(3-(pentylamino)phenyl)propylcarbamate (0.15 g, 0.45
mmol) following method described in Example 1 gave Example 13
hydrochloride as a white solid. Yield (0.10 g, 95%): .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 7.52-7.61 (m, 3H), 7.38-7.42 (m, 1H),
4.92 (dd, J=9.2, 3.6 Hz, 1H), 3.36-3.40 (m, 2H), 3.08-3.18 (m, 2H),
1.92-2.12 (m, 2H), 1.70-1.80 (m, 2H), 1.34-1.46 (m, 4H), 0.90-0.98
(m, 3H).
Example 14
Preparation of 3-amino-1-(3-(pentylamino)phenyl)propan-1-one
##STR00311##
[1159] 3-Amino-1-(3-(pentylamino)phenyl)propan-1-one was prepared
following the method used in Examples 13 and 12
[1160] Step 1: Oxidation of tert-butyl
3-hydroxy-3-(3-(pentylamino)phenyl)propylcarbamate following the
method used in Example 12 gave tert-butyl
3-oxo-3-(3-(pentylamino)phenyl)propylcarbamate as a light yellow
oil which was directly used in next reaction without further
purification. Yield (0.05 g, 50%).
[1161] Step 2: Deprotection tert-butyl
3-oxo-3-(3-(pentylamino)phenyl)propylcarbamate (0.05 g, 0.15 mmol)
following the method used in Example 12 gave Example 14 as a white
solid. Yield (0.03 g, 85%); .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.14-8.19 (m, 2H), 7.73-7.82 (m, 2H), 3.53 (t, J=6.0 Hz,
2H), 3.43 (t, J=7.6 Hz, 2H), 3.36 (t, J=5.2 Hz, 2H), 1.73-1.83 (m,
2H), 1.35-1.46 (m, 4H), 0.93 (t, J=7.2 Hz, 3H).
Example 15
Preparation of
N-(3-(3-amino-1-hydroxypropyl)phenyl)cyclohexanecarboxamide
##STR00312##
[1163] N-(3-(3-Amino-1-hydroxypropyl)phenyl)cyclohexanecarboxamide
was prepared following the method shown in Scheme 10.
##STR00313##
[1164] Step 1: Reduction of nitrile 12 following the method
described in Example 11 gave crude amine 34 hydrochloride which was
used directly in next step without further purification.
[1165] Step 2: To a suspension of crude amine salt 34 (0.94 g, 4.64
mmol) in dichloromethane (15 mL) and TEA (0.7 ml, 5.0 mmol) was
added dropwise a solution of Boc.sub.2O (1.0 g, 4.64 mmol) in DCM
at room temperature. The reaction mixture was stirred at room
temperature for 2 hr, washed with aqueous NH.sub.4Cl, dried over
anhydrous Na.sub.2SO.sub.4 and concentrated under reduced pressure.
Purification by flash chromatography (65% to 75% EtOAc-hexanes
gradient) gave carbamate 35 as a colorless oil. Yield (0.8 g, 64%);
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 6.90 (t, J=7.6 Hz, 1H),
6.69 (t, J=5.6 Hz, 1H), 6.51 (t, J=1.2 Hz, 1H), 6.36-6.41 (m, 2H),
4.97 (d, J=4.0 Hz, 1H), 4.92 (br.s, 2H), 4.34 (q, J=4.0 Hz, 1H),
2.88-2.94 (m, 2H), 1.60 (q, J=6.8 Hz, 2H), 1.34 (s, 9H).
[1166] Step 3: To a solution of carbamate 35 (0.43 g. 1.61 mmol),
TEA (0.24 ml, 1.76 mmol) in THF was added dropwise a solution of
cyclohexanecarbonyl chloride (36) (0.2 ml, 1.61 mmol) in THF at
0.degree. C. The resulting mixture was allowed to warm to room
temperature, stirred for 1 hr and then a mixture of 25%
NH.sub.4Cl-0.5N HCl (20 ml) was added. Organic layer was separated,
dried over anhydrous Na.sub.2SO.sub.4 and concentrated under
reduced pressure. Purification by flash chromatography (65 to 70%
EtOAc-hexanes gradient) gave tert-butyl
3-(3-(cyclohexanecarboxamido)phenyl)-3-hydroxypropylcarbamate (37)
as a colorless oil. Yield (0.5 g, 82%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.72 (s, 1H), 7.54 (s, 1H), 7.44 (dd, J=8.0,
1.2 Hz, 1H), 7.17 (t, J=8.0 Hz, 1H), 6.92 (d, J=7.6 Hz, 1H), 6.72
(t, J=4.8 Hz, 1H), 5.15 (d, J=4.4 Hz, 1H), 4.47 (q, J=4.4 Hz, 1H),
2.94 (q, J=6.4 Hz, 2H), 2.24-2.33 (m, 1H), 1.50-1.79 (m, 7H),
1.14-1.93 (m, 14H).
[1167] Step 4: Deprotection of carbamate 37 following method
described in Example 1 gave Example 17 hydrochloride as a white
solid. Yield (0.75 g, 91%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.82 (s, 1H), 7.82 (m, 3H), 7.66 (s, 1H), 7.40 (d, J=7.2
Hz, 1H), 7.20 (t, J=8.0 Hz, 1H), 6.94 (d, J=7.6 Hz, 1H), 4.60 (dd,
J=8.0, 4.8 Hz, 1H), 2.78-2.89 (m, 2H), 2.26-2.36 (m, 1H), 1.58-1.86
(m, 7H), 1.13-1.45 (m, 5H).
Example 16
Preparation of
N-(3-(3-aminopropanoyl)phenyl)cyclohexanecarboxamide
##STR00314##
[1169] N-(3-(3-Aminopropanoyl)phenyl)cyclohexanecarboxamide was
prepared following the method used in Example 15.
[1170] Step 1: Oxidation of tert-butyl
3-(3-(cyclohexanecarboxamido)phenyl)-3-hydroxypropylcarbamate (37)
following the method used in Example 12 except PCC was used in lieu
of MnO.sub.2 gave tert-butyl
3-(3-(cyclohexanecarboxamido)phenyl)-3-oxopropylcarbamate as a
white solid. Yield (0.36 g, 91%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.97 (s, 1H), 8.15-8.18 (m, 1H), 7.82 (dd,
J=1.2, 8.0 Hz, 1H), 7.57 (d, J=8.0 Hz, 1H), 7.40 (t, J=8.0 Hz, 1H),
6.79 (br.t, 1H), 3.24 (q, J=6.0 Hz, 2H), 3.08 (t, J=6.4 Hz, 2H),
2.25-2.35 (m, 1H), 1.69-1.82 (m, 4H), 1.58-1.66 (m, 1H), 1.33 (s,
9H), 1.10-1.44 (m, 5H).
[1171] Step 2: tert-Butyl
3-(3-(cyclohexanecarboxamido)phenyl)-3-oxopropylcarbamate was
deprotected following the method used in Example 15 to give Example
16 hydrochloride as a white solid. Yield (0.060 g, 81%); .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 8.05-8.07 (m, 2H), 7.65-7.72 (m,
2H), 3.48 (t, J=6.0 Hz, 2H), 3.34-3.37 (m, 2H), 3.27 (t, J=6.8 Hz,
2H), 1.68-1.91 (m, 6H), 1.22-1.38 (m, 3H), 1.04-1.16 (m, 2H).
Example 17
Preparation of N-(3-(3-amino-1-hydroxypropyl)phenyl)pentanamide
##STR00315##
[1173] N-(3-(3-Amino-1-hydroxypropyl)phenyl)pentanamide was
prepared following the method described below.
[1174] Step 1: To a solution of carbamate 35 (0.43 g. 1.61 mmol),
TEA (0.24 ml, 1.76 mmol) in THF (20 ml) was added pentanoyl
chloride (0.19 ml, 1.55 mmol) in THF (10 ml) at 0.degree. C.
dropwise. The resulting mixture was allowed to room temperature and
stirred for 1 hr and then added NH.sub.4Cl--HCl.aq (0.5 N, 20 ml).
Layers were separated, dried over anhydrous Na.sub.2SO.sub.4 and
concentrated under reduced pressure. Purification by flash
chromatography (65 to 70% EtOAc-hexanes gradient) gave tert-butyl
3-hydroxy-3-(3-pentanamidophenyl)propylcarbamate as a colorless
oil. Yield (0.5 g, 92%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.79 (s, 1H), 7.51 (s, 1H), 7.45 (d, J=8.0 Hz, 1H), 7.18
(t, J=8.0 Hz, 1H), 6.92 (d, J=8.0 Hz, 1H), 6.73 (t, J=5.6 Hz, 1H),
5.16 (d, J=4.4 Hz, 1H), 4.67 (q, J=4.8 Hz, 1H), 2.94 (q, J=6.4 Hz,
2H), 2.26 (t, J=7.2 Hz, 2H), 1.63 (q, J=7.2 Hz, 2H), 1.50-1.58 (m,
2H), 1.26-1.34 (m, 11H), 0.89 (t, J=8.4 Hz, 3H).
[1175] Step 2: Deprotection of tert-butyl
3-hydroxy-3-(3-pentanamidophenyl)propylcarbamate following the
method used in Example 15 gave Example 17 as a white solid. Yield
(0.77 g, 95%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.94 (s,
1H), 7.94 (br s, 3H), 7.62 (s, 1H), 7.44 (d, J=7.6 Hz, 1H), 7.20
(t, J=7.6 Hz, 1H), 6.95 (d, J=7.2 Hz, 1H), 4.60 (t, J=5.6 Hz, 1H),
2.28 (t, J=7.6 Hz, 2H), 2.74-2.88 (m, 2H), 1.75-1.90 (m, 2H),
1.50-1.58 (m, 2H), 1.26-1.38 (m, 2H), 0.86 (t, J=7.2 Hz, 3H).
Example 18
Preparation of N-(3-(3-aminopropanoyl)phenyl)pentanamide
##STR00316##
[1177] N-(3-(3-Aminopropanoyl)phenyl)pentanamide was prepared
following the method used in Examples 17, 16.
[1178] Step 1: Oxidation of tert-butyl
3-hydroxy-3-(3-pentanamidophenyl)propylcarbamate following the
method used in Example 16 gave tert-butyl
3-oxo-3-(3-pentanamidophenyl)propylcarbamate as a light yellow oil.
Yield (0.34 g, 84%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
10.04 (s, 1H), 8.13 (s, 1H), 7.12 (d, J=8.8 Hz, 1H), 7.58 (d, J=7.6
Hz, 1H), 7.41 (t, J=8.0 Hz, 1H), 6.79 (d, J=5.6 Hz, 1H), 3.23 (q,
J=6.0 Hz, 2H), 3.09 (t, J=6.8 Hz, 2H), 2.29 (t, J=7.6 Hz, 2H),
1.52-1.60 (m, 2H), 1.26-1.36 (m, 11H), 0.87 (t, J=7.2 Hz, 3H).
[1179] Step 2: Deprotection tert-butyl
3-oxo-3-(3-pentanamidophenyl)propylcarbamate following the method
used in Example 16 gave Example 18 as a white solid. Yield (0.09 g,
90%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.18 (s, 1H),
8.29 (t, J=2.0 Hz, 1H), 7.78-7.82 (m, 4H), 7.60-7.63 (m, 1H), 7.46
(t, J=8.0 Hz, 1H), 3.53 (t, J=7.2 Hz, 2H), 3.12 (q, J=5.6 Hz, 2H),
3.21 (t, J=7.6 Hz, 2H), 1.52-1.60 (m, 2H), 1.26-1.36 (m, 2H), 0.88
(t, J=7.6 Hz, 3H).
Example 19
Preparation of
3-(3-amino-1-fluoropropyl)-N-(cyclohexylmethyl)aniline
##STR00317##
[1181] 3-(3-Amino-1-fluoropropyl)-N-(cyclohexylmethyl)aniline is
prepared following the method described below.
[1182] Step 1: A mixture of alcohol 32 and DAST are stirred at
-78.degree. C. until no starting material is seen by TLC. The
reaction mixture is then quenched by addition of aqueous
NH.sub.4Cl. Layers are separated and aqueous layer is additionally
extracted with EtOAc. Combined organic layers are washed with
brine, dried over anhydrous MgSO.sub.4 and concentrated under
reduced pressure. Purification by flash chromatography gives
3-(3-(cyclohexylmethylamino)phenyl)-3-fluoropropanenitrile.
[1183] Step 2:
3-(3-(Cyclohexylmethylamino)phenyl)-3-fluoropropanenitrile is
reduced with BH.sub.3-Me.sub.2S following the method used in
Example 11 to give Example 19.
Example 20
Preparation of
N-(3-(3-aminopropanoyl)phenyl)cyclohexanesulfonamide
##STR00318##
[1185] N-(3-(3-Aminopropanoyl)phenyl)cyclohexanesulfonamide was
prepared following the method shown in Scheme 11.
##STR00319##
[1186] Step 1: To a solution of Example 5 (0.26 g, 0.83 mmol) in
DCM (10 mL) was added Boc.sub.2O (0.22 g, 1.0 mmol). The reaction
mixture was stirred at room temperature for 18 hrs and concentrated
under reduced pressure. Carbamate 38 was used in the next step
without purification.
[1187] Step 2: To a solution of alcohol 38 (approx. 0.83 mmol) in
dichloromethane (15 mL) was added Des Martin periodinane (0.4 g,
0.92 mmol). The mixture was stirred for 1 h at room temp, washed
with brine and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated under reduced pressure. Purification by flash
chromatography (40 to 55% EtOAc-hexanes gradient) gave ketone 39 as
a light yellow oil. Yield (0.06 g, 18%); .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.82 (m, 1H), 7.70 (d, J=7.6 Hz, 1H), 7.41-7.50
(m, 2H), 3.42 (t, J=6.8 Hz, 2H), 3.17 (t, J=6.4 Hz, 2H), 2.08-2.16
(m, 2H), 1.80-1.88 (m, 2H), 1.61-1.69 (m, 1H), 1.46-1.58 (m, 2H),
1.41 (s, 9H), 1.15-1.30 (m, 2H).
[1188] Step 3: To a solution of ketone 39 (0.06 g. 0.14 mmol) in
EtOAc was added HCl (5 ml of a 6.9 M solution in EtOH, 34.5 mmol).
The reaction mixture was stirred at room temperature for 3 hrs and
concentrated under reduced pressure to give a Example 20 as a white
solid. Yield (0.049 g, 99%); .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.91 (t, J=2.0 Hz, 1H), 7.73-7.75 (m, 1H), 7.46-7.52 (m,
2H), 3.48 (t, J=6.0 Hz, 2H), 3.35 (t, J=6.0 Hz, 2H), 2.94-3.04 (m,
1H), 2.08-2.11 (m, 2H), 1.79-1.83 (m, 2H), 1.61-1.67 (m, 1H),
1.44-1.58 (m, 2H), 1.10-1.28 (m, 3H).
Example 21
Preparation of
N-(3-(3-amino-1-hydroxypropyl)phenyl)butane-1-sulfonamide
##STR00320##
[1190] N-(3-(3-Amino-1-hydroxypropyl)phenyl)butane-1-sulfonamide
was prepared following the method used in Example 5.
[1191] Step 1: Coupling of aniline 12 with butane-1-sulfonyl
chloride (0.47 ml, 3.5 mmol) gave
N-(3-(2-cyano-1-hydroxyethyl)phenyl)butane-1-sulfonamide as a light
yellow oil. Yield (0.80 g, 89%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.76 (s, 1H), 7.24-7.29 (m, 2H), 7.06-7.09
(m, 2H), 5.95 (d, J=4.4 Hz, 1H), 4.83 (q, J=4.4 Hz, 1H), 3.05 (t,
J=8.0 Hz, 2H), 2.79 (ddd, J=24.8, 16.8, 6.4 Hz, 2H), 1.56-1.64 (m,
2H), 1.26-1.35 (m, 2H), 0.79 (t, J=8.4 Hz, 3H).
[1192] Step 2: Reduction of
N-(3-(2-cyano-1-hydroxyethyl)phenyl)butane-1-sulfonamide with
BH.sub.3-Me.sub.2S gave Example 21 as a light yellow oil. Yield
(0.76 g, 93%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.15-7.22 (m, 2H), 6.96-7.02 (m, 2H), 4.59 (t, J=6.8 Hz, 1H), 3.00
(t, J=8.4 Hz, 2H), 2.56-21.68 (m, 2H), 1.56-1.64 (m, 4H), 1.26-1.36
(m, 2H), 0.79 (t, J=7.6 Hz, 3H).
Example 22
Preparation of
N-(3-(3-aminopropanoyl)phenyl)butane-1-sulfonamide
##STR00321##
[1194] N-(3-(3-Aminopropanoyl)phenyl)butane-1-sulfonamide was
prepared following the method used in Example 20.
[1195] Step 1: Protection of Example 21 with Boc.sub.2O following
the method used in Example 20 gave tert-butyl
3-(3-(butylsulfonamido)phenyl)-3-hydroxypropylcarbamate as a
colorless oil. Yield (0.18 g, 15%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.30-7.38 (m, 2H), 7.18 (s, 1H), 7.06-7.10
(m, 1H), 6.76 (t, J=5.6 Hz, 1H), 5.31 (d, J=4.8 Hz, 1H), 4.57 (q,
J=5.2 Hz, 1H), 3.69 (t, J=8.0 Hz, 2H), 2.90-2.98 (m, 2H), 1.60-1.78
(m, 4H), 1.34-1.45 (m, 20H), 0.90 (t, J=7.2 Hz, 3H).
[1196] Step 2: Oxidation of tert-butyl
3-(3-(butylsulfonamido)phenyl)-3-hydroxypropylcarbamate by PCC
following the method used in Example 18 gave tert-butyl
3-(3-(butylsulfonamido)phenyl)-3-oxopropylcarbamate as white solid:
Yield (0.18 g, 41%); .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
8.01-8.04 (m, 1H), 7.82-7.84 (m, 1H), 7.55 (t, J=8.0 Hz, 1H),
7.49-7.52 (m, 1H), 5.86-5.64 (m, 1H), 3.71-3.75 (m, 2H), 3.43 (q,
J=6.0 Hz, 2H), 3.21 (t, J=6.8 Hz, 2H), 1.82-1.90 (m, 2H), 1.48-1.58
(m, 2H), 1.41-1.44 (m, 18H), 0.99 (t, J=7.2 Hz, 3H).
[1197] Step 3: Deprotection of tert-butyl
3-(3-(butylsulfonamido)phenyl)-3-oxopropylcarbamate following the
method used in Example 20 gave Example 22 as a white solid. Yield
(0.05 g, 97%); .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.92 (t,
J=2.0 Hz, 1H), 7.76-7.78 (m, 1H), 7.44-7.52 (m, 2H), 3.43 (t, J=6.0
Hz, 2H), 3.30-3.38 (m, 2H), 3.07-3.11 (m, 2H), 1.70-1.79 (m, 2H),
1.36-1.46 (m, 2H), 0.88 (t, J=7.2 Hz, 3H).
Example 23
Preparation of
(E)-3-(3-aminoprop-1-enyl)-N-(cyclohexylmethyl)aniline
##STR00322##
[1199] (E)-3-(3-Aminoprop-1-enyl)-N-(cyclohexylmethyl)aniline was
prepared following the method described below.
[1200] Step 1: Cyclohexanecarbonyl chloride (0.74 g, 6.97 mmol) was
added to a mixture of 3-bromoaniline (1.0 g, 5.8 mmol), TEA (1.07
mL, 7.55 mmol) and DMAP (cat.) in THF with stirring at 0.degree. C.
over 10 min. Stirring was continued for another 30 min and quenched
with saturated NaHCO.sub.3. The product was extracted with ethyl
acetate. Combined organic layers were concentrated under reduced
pressure to give a residue which was triturated with pentane to
give N-(3-bromophenyl)cyclohexanecarboxamide as an off-white solid.
Yield (1.3 g, 79%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.96 (s, 1H), 7.97 (s, 1H), 7.49 (d, J=8.0 Hz, 1H), 7.26-7.18 (m,
2H), 2.33-2.26 (m, 1H), 1.76 (t, J=14.0 Hz, 4H), 1.65 (d, J=10.4
Hz, 1H), 1.43-1.34 (m, 2H), 1.30-1.12 (m, 3H).
[1201] Step 2: Reduction of N-(3-bromophenyl)cyclohexanecarboxamide
with BH.sub.3-Me.sub.2S following the method used in Example 11
gave 3-bromo-N-(cyclohexylmethyl)aniline as a colorless oil. Yield
(1.0 g, 80%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 6.96 (t,
J=8.0 Hz, 1H), 6.68 (s, 1H), 6.60 (d, J=7.6 Hz, 1H), 6.53 (d, J=8.4
Hz, 1H), 5.93 (t, J=5.6 Hz, 1H), 2.81 (t, J=6.2 Hz, 2H), 1.78 (d,
J=12.8 Hz, 2H), 1.69-1.61 (m, 3H), 1.53-1.46 (m, 1H), 1.24-1.08 (m,
3H), 0.99-0.77 (m, 2H).
[1202] Step 3: Trifluoroacetic anhydride (0.75 ml, 4.49 mmol) was
added to a mixture of 3-bromo-N-(cyclohexylmethyl)aniline (1.0 g,
3.74 mmol), TEA (0.8 ml) in CH.sub.2Cl.sub.2 at 0.degree. C. in 10
min time. The reaction mixture was stirred for 30 min at room
temperature and partitioned between saturated NaHCO.sub.3 and
extracted with EtOAc three times. Combined organic layers were
concentrated under reduced pressure to give
N-(3-bromophenyl)-N-(cyclohexylmethyl)-2,2,2-trifluoroacetamide as
a colorless liquid. Yield (1.0 g, 74%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.56 (d, J=8.0 Hz, 1H), 7.39 (s, 1H), 7.31 (t,
J=8.0 Hz, 1H), 7.18 (d, J=8.0 Hz, 1H), 3.61 (d, J=6.0 Hz, 2H),
1.72-1.64 (m, 5H), 1.51 (bs, 1H), 1.18-1.16 (m, 3H), 1.06-1.01 (m,
2H).
[1203] Step 4:
N-(3-Bromophenyl)-N-(cyclohexylmethyl)-2,2,2-trifluoroacetamide
(1.0 g, 2.74 mmol), N-allyl-2,2,2-trifluoroacetamide (0.5 g, 3.29
mmol), tri-O-tolylphosphine (0.08 g, 0.27 mmol) and triethylamine
(2 mL, 13.7 mmol) was added to DMF and the mixture was flushed with
argon for 15 min. Pd(OAc).sub.2 (0.06 g, 0.27 mmol) was charged to
the reaction mixture which was stirred at 90.degree. C. for 2 h.
The reaction mixture was cooled and partitioned between ethyl
acetate and water. Organic layer was washed thoroughly with water,
dried over anhydrous sodium sulfate and concentrated under reduced
pressure. Purification by column chromatography (100-200 silica
mesh, 5% to 10% EtOAc in hexane) gave
(E)-N-(cyclohexylmethyl)-2,2,2-trifluoro-N-(3-(3-(2,2,2-trifluoroacetamid-
o)prop-1-enyl)phenyl)acetamide 5 as a colorless semi solid. Yield
(0.4 g, 33%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.43-7.41
(m, 2H), 7.18 (s, 1H), 7.13 (d, J=6.8 Hz, 1H), 6.61 (d, J=16.0 Hz,
1H), 6.48 (bs, 1H), 6.24-6.16 (m, 1H), 4.17 (t, J=6.0 Hz, 2H), 3.62
(bs, 2H), 1.73-1.65 (m, 5H), 1.56-1.48 (m, 1H), 1.21-1.14 (m, 3H),
1.07-0.99 (m, 2H).
[1204] Step 5: A mixture of
(E)-N-(cyclohexylmethyl)-2,2,2-trifluoro-N-(3-(3-(2,2,2-trifluoroacetamid-
o)prop-1-enyl)phenyl)acetamide (0.2 g, 0.45 mmol) and
K.sub.2CO.sub.3 (0.19 g, 1.37 mmol) in MeOH:H.sub.2O was stirred at
room temperature for 24 h and then at 50.degree. C. for 16 h. The
solvent was removed under reduced pressure. Purification by column
chromatography (5% to 10% MeOH-CH.sub.2Cl.sub.2 gradient) gave
3-(3-aminoprop-1-enyl)-N-(cyclohexylmethyl)aniline as pale brown
semi-solid. Yield (0.06 g, 54%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.99 (t, J=8.0 Hz, 1H), 6.54-6.53 (m, 2H),
6.44 (d, J=7.2 Hz, 1H), 6.42 (d, J=16.0 Hz, 1H), 6.22-6.15 (m, 1H),
5.58 (t, J=5.8 Hz, 1H), 3.34 (d, J=5.2 Hz, 2H), 2.83 (t, J=5.8 Hz,
2H), 1.80-1.77 (m, 2H), 1.70-1.56 (m, 3H), 1.54-1.49 (m, 1H),
1.18-1.12 (m, 3H), 0.97-0.91 (m, 2H); RP-HPLC (Method 4)
t.sub.R=5.30 min, 96.10% (AUC); ESI MS m/z 245.26 [M+H].sup.+.
Example 24
Preparation of
3-(3-aminoprop-1-ynyl)-N-(cyclohexylmethyl)aniline
##STR00323##
[1206] 3-(3-Aminoprop-1-ynyl)-N-(cyclohexylmethyl)aniline was
prepared following the method used in Example 23 and as described
below.
[1207] Step 1: Triethylamine (45 mL) was added to a mixture of
N-(3-bromophenyl)-N-(cyclohexylmethyl)-2,2,2-trifluoroacetamide
(3.8 g, 10.4 mmol), tert-butyl prop-2-ynylcarbamate (2.42 g, 15.6
mmol), Pd(Ph.sub.3P).sub.4 (0.6 g, 0.52 mmol) and CuI (0.1 g, 0.52
mmol) and flushed for 15 min with argon. The reaction mixture was
stirred for 16 h at 90.degree. C. The reaction mixture was cooled,
diluted with ethyl acetate and filtered through Celite bed and the
filtrate was concentrated under reduced pressure. Purification by
column chromatography (100-200 silica mesh 5% to 10% EtOAc-hexane)
gave tert-butyl
3-(3-(N-(cyclohexylmethyl)-2,2,2-trifluoroacetamido)phenyl)prop-2-ynylcar-
bamate as a yellow semi-solid. Yield (2.1 g, 50%); .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 7.49-7.48 (m, 2H), 7.43-7.41 (m, 2H),
3.99 (bs, 2H), 3.58 (bs, 2H), 1.64-1.57 (m, 6H), 1.39 (s, 9H),
1.19-1.12 (m, 3H), 0.95-0.89 (m, 2H).
[1208] Step 2: A mixture of 50% CF.sub.3COOH in DCM (20 mL) and
tert-butyl
3-(3-(N-(cyclohexylmethyl)-2,2,2-trifluoroacetamido)phenyl)prop-2-ynylcar-
bamate (1.6 g, 4.67 mmol) was initially stirred at 0.degree. C. and
stirring continued at room temperature for 3 h. The reaction
mixture was evaporated to dryness and triturated with pentane to
give Example 24 trifluoroacetate as a brown oil. Yield (0.46 g,
54%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.29 (br.s, 3H),
7.08 (t, J=8.0 Hz, 1H), 6.65 (d, J=8.8 Hz, 1H), 6.60-6.59 (m, 2H),
3.98-3.96 (m, 2H), 2.83 (d, J=6.4 Hz, 2H), 1.78-1.75 (m, 2H),
1.70-1.63 (m, 3H), 1.52-1.48 (m, 1H), 1.24-1.15 (m, 3H), 0.99-0.88
(m, 2H). RP-HPLC (Method 6) t.sub.R=6.17 min, 99.70% (AUC); ESI MS
m/z 243.23[M+H].sup.+.
Example 25
Preparation of
(E)-N-(3-(3-aminoprop-1-enyl)phenyl)cyclohexanecarboxamide
##STR00324##
[1210] (E)-N-(3-(3-Aminoprop-1-enyl)phenyl)cyclohexanecarboxamide
is prepared following the method used in Examples 33 and 15.
[1211] Step 1: Acylation of
(E)-N-(3-(3-aminophenyl)allyl)-2,2,2-trifluoroacetamide following
the method used in Example 15 gives
(E)-N-(3-(3-(2,2,2-trifluoroacetamido)prop-1-enyl)phenyl)cyclohexanecarbo-
xamide.
[1212] Step 2: Deprotection of
(E)-N-(3-(3-(2,2,2-trifluoroacetamido)prop-1-enyl)phenyl)cyclohexanecarbo-
xamide following the method used in Example 33 gives Example
25.
Example 26
Preparation of
N-(3-(3-aminoprop-1-ynyl)phenyl)cyclohexanecarboxamide
##STR00325##
[1214] N-(3-(3-Aminoprop-1-ynyl)phenyl)cyclohexanecarboxamide is
prepared following the method used in Examples 24 and 25.
[1215] Step 1: Acylation of tert-butyl
3-(3-aminophenyl)prop-2-ynylcarbamate following the method used in
Example 25 gives tert-butyl
3-(3-(cyclohexanecarboxamido)phenyl)prop-2-ynylcarbamate.
[1216] Step 2: Deprotection of tert-butyl
3-(3-(cyclohexanecarboxamido)phenyl)prop-2-ynylcarbamate following
the method used in Example 24 gives Example 26 hydrochloride.
Example 27
Preparation of
(E)-N-(3-(3-aminoprop-1-enyl)phenyl)cyclohexanesulfonamide
##STR00326##
[1218] (E)-N-(3-(3-Aminoprop-1-enyl)phenyl)cyclohexanesulfonamide
is prepared following the method used in Example 33 and 5.
[1219] Step 1: Sulfonation of
(E)-N-(3-(3-aminophenyl)allyl)-2,2,2-trifluoroacetamide following
the method used in Example 5 gives
(E)-N-(3-(3-(cyclohexanesulfonamido)phenyl)allyl)-2,2,2-trifluoroacetamid-
e.
[1220] Step 2: Deprotection of
(E)-N-(3-(3-(cyclohexanesulfonamido)phenyl)allyl)-2,2,2-trifluoroacetamid-
e following the method used in Example 33 gives Example 27.
Example 28
Preparation of
N-(3-(3-aminoprop-1-ynyl)phenyl)cyclohexanesulfonamide
##STR00327##
[1222] N-(3-(3-Aminoprop-1-ynyl)phenyl)cyclohexanesulfonamide is
prepared following the method used in Examples 24 and 5.
[1223] Step 1: Sulfonation of tert-butyl
3-(3-aminophenyl)prop-2-ynylcarbamate following the method used in
Example 5 gives tert-butyl
3-(3-(cyclohexanesulfonamido)phenyl)prop-2-ynylcarbamate.
[1224] Step 2: Deprotection of tert-butyl
3-(3-(cyclohexanesulfonamido)phenyl)prop-2-ynylcarbamate following
the method used in Example 24 gives Example 28 hydrochloride.
Example 29
Preparation of
(E)-1-((3-(3-aminoprop-1-enyl)phenylamino)methyl)cyclohexanol
##STR00328##
[1226]
(E)-1-((3-(3-Aminoprop-1-enyl)phenylamino)methyl)cyclohexanol was
prepared following the method described below and in Example
33.
[1227] Step 1: A mixture of
(E)-N-(3-(3-aminophenyl)allyl)-2,2,2-trifluoroacetamide (0.8 g,
3.28 mmol) and 1-oxaspiro[2.5]octane (0.55 g, 4.91 mmol) in
EtOH:H.sub.2O(9:1) was stirred under reflux for 36 hrs and
concentrated under reduced pressure. Purification by column
chromatography (20% to 30% EtOAc-hexanes gradient) gave
(E)-2,2,2-trifluoro-N-(3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)ally-
l)acetamide as an off-white solid. Yield (0.5 g, 43%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.71 (s, 1H), 7.00 (t, J=8.0 Hz,
1H), 6.67 (s, 1H), 6.55 (t, J=8.8 Hz, 2H), 6.41 (d, J=16.0 Hz, 1H),
6.61-6.09 (m, 1H), 5.22 (t, J=5.2 Hz, 1H), 4.20 (s, 1H), 3.95 (t,
J=5.2 Hz, 2H), 2.94 (d, J=5.6, 2H), 1.61-1.49 (m, 5H), 1.41-1.36
(m, 4H), 1.25-1.19 (m, 1H).
[1228] Step 2: A mixture of
(E)-2,2,2-trifluoro-N-(3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)ally-
l)acetamide (0.5 g, 1.14 mmol) and potassium carbonate (0.29 g, 2.1
mmol) in methanol:water (1:1) was stirred at room temperature for
24 hrs. The solvent was evaporated under reduced pressure.
Purification by column chromatography (5% to 10% MeOH-DCM gradient)
gave Example 29 as an off-white solid. Yield (0.11 g, 36%); .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 7.72 (bs, 2H), 7.03 (t, J=7.8
Hz, 1H), 6.66 (s, 1H), 6.60-6.56 (m, 3H), 6.18-6.11 (m, 1H), 5.34
(t, J=5.8 Hz, 1H), 4.23 (s, 1H), 3.57 (d, J=6.4 Hz, 2H), 2.94 (d,
J=6.0 Hz, 2H), 1.62-1.50 (m, 5H), 1.41-1.38 (m, 4H), 1.23-1.18 (m,
1H); RP-HPLC (Method 3) t.sub.R=3.55 min, 99.20% (AUC); ESI MS m/z
261.29 [M+H].sup.+.
Example 30
Preparation of
1-((3-(3-aminoprop-1-ynyl)phenylamino)methyl)cyclohexanol
##STR00329##
[1230] 1-((3-(3-Aminoprop-1-ynyl)phenylamino)methyl)cyclohexanol is
prepared following the method used in Examples 24 and 29.
[1231] Step 1: 2,2,2-Trifluoro-N-(prop-2-ynyl)acetamide (3.4 g,
22.2 mmol), 1-bromo-3-nitrobenzene (14) (3.0 g, 14.85 mmol),
Pd(Ph.sub.3P).sub.4 (0.85 g, 0.74 mmol) and CuI (0.14 g, 0.74 mmol)
was added to triethylamine (30 mL) and the mixture was flushed with
argon for 15 min. The reaction mixture was stirred at 90.degree. C.
for 16 h, cooled and diluted with ethyl acetate. The mixture was
filtered through Celite and the filtrate was concentrated under
reduced pressure. Purification by column chromatography (100-200
silica mesh, 15% to 20% EtOAc in hexane) gave
2,2,2-trifluoro-N-(3-(3-nitrophenyl)prop-2-ynyl)acetamide as a
brown semi-solid. Yield (1.95 g, 48%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.22 (s, 1H), 8.16 (d, J=8.0 Hz, 1H), 1.79
(d, J=7.6 Hz, 1H), 7.66 (t, J=8.0 Hz, 1H), 7.34 (s, 1H), 4.38 (s,
2H).
[1232] Step 2: Tin(II) chloride dihydrate was added (6.5 g, 28.67
mmol) to a solution of
2,2,2-trifluoro-N-(3-(3-nitrophenyl)prop-2-ynyl)acetamide (1.95 g,
7.16 mmol) in ethanol and the reaction mixture was stirred under
reflux overnight. The mixture was concentrated under reduced
pressure to give dark brown viscous liquid which was partitioned
between saturated aqueous NaHCO.sub.3 and EtOAc. Organic layer was
dried over anhydrous sodium sulfate and concentrated under reduced
pressure to give
N-(3-(3-aminophenyl)prop-2-ynyl)-2,2,2-trifluoroacetamide as brown
oil. Yield (0.75 g, 43%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.28 (s, 1H), 6.94 (t, J=7.8 Hz, 1H), 6.44-6.37 (m, 3H),
5.09 (bs, 2H), 3.99 (s, 2H).
[1233] Step 3: A mixture of
N-(3-(3-aminophenyl)prop-2-ynyl)-2,2,2-trifluoroacetamide (0.75 g,
3.09 mmol) and 1-oxaspiro[2.5]octane (1.2 g, 9.2 mmol) in
EtOH:H.sub.2O (9:1) was stirred under reflux for 36 h. The reaction
mixture was concentrated under reduced pressure. Purification by
column chromatography (20% to 30% EtOAc-hexanes gradient) gave
2,2,2-trifluoro-N-(3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)prop-2-y-
nyl)acetamide as an off white solid. Yield (0.48 g, 43%); .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 7.28 (s, 1H), 7.00 (t, J=7.6
Hz, 1H), 6.53-6.51 (m, 2H), 6.39 (d, J=7.6 Hz, 1H), 5.28 (t, J=5.6
Hz, 1H), 4.18 (s, 1H), 4.01 (s, 2H), 2.88 (d, J=5.6 Hz, 2H),
1.60-1.47 (m, 4H), 1.39-1.34 (m, 4H), 1.22-1.14 (m, 2H).
[1234] Step 4: Deprotection of
2,2,2-trifluoro-N-(3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)prop-2-y-
nyl)acetamide gives Example 30.
Example 31
Preparation of 3-(3-aminopropyl)-N-(cyclopentylmethyl)aniline
##STR00330##
[1236] 3-(3-Aminopropyl)-N-(cyclopentylmethyl)aniline was prepared
following the method shown in Scheme 12.
##STR00331##
[1237] Step 1: A mixture of nitrobenzene 40 (0.5 g, 1.6 mmol) and
cyclopentanecarbaldehyde (0.15 ml, 1.6 mmol) in EtOAc was degassed
and saturated with argon. 10% Pd/C (0.40 g) was added to this
solution and the resulting mixture was stirred under H.sub.2 at 1
atm for 3 hrs. The reaction mixture was filtered through Celite,
concentrated under reduced pressure. Purification by flash
chromatography (40% to 50% EtOAc-hexanes gradient) gave aniline 41
as a yellow semi-solid. Yield (0.4 g, 68%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.86-7.80 (m, 4H), 6.90 (t, J=8.0 Hz, 1H),
6.36 (s, 1H), 6.33 (d, J=5.6 Hz, 2H), 5.40 (t, J=5.6 Hz, 1H), 3.59
(t, J=7.2 Hz, 2H), 2.86 (t, J=6.4 Hz, 2H),), 2.50-2.45 (m, 2H),
2.09 (quintet, J=7.6 Hz, 1H), 1.86 (quintet, J=7.6 Hz, 2H),
1.76-1.72 (m, 2H), 1.57-1.47 (m, 4H), 1.23-1.08 (m, 2H).
[1238] Step 2: A mixture of alkylphthalimide 41 (350 mg, 0.96 mmol)
and hydrazine hydrate (0.1 ml) in methanol was stirred at room
temperature for 24 hours. The solvent was evaporated under reduced
pressure. Purification by flash chromatography (5% to 6% MeOH-DCM
gradient) gave Example 31 as a colourless semi-solid. Yield (0.16
g, 71%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 6.95 (t, J=8.0
Hz, 1H), 6.38 (bs, 2H), 6.33 (d, J=7.6 Hz, 1H), 5.47 (t, J=5.2 Hz,
1H), 3.5 (bs, 2H), 2.87 (t, J=6.4 Hz, 2H), 2.66 (t, J=7.2 Hz, 2H),
2.50-2.45 (m, 2H), 2.12 (quintet, J=7.6 Hz, 1H), 1.75-1.67 (m, 4H),
1.58-1.50 (m, 4H), 1.26-1.21 (m, 2H). RP-HPLC (Method 3)
t.sub.R=5.18 min, 97.03% (AUC); ESI MS m/z 233.27 [M+H]+.
Example 32
Preparation of 3-(3-aminopropyl)-N-(2-propylpentyl)aniline
##STR00332##
[1240] 3-(3-Aminopropyl)-N-(2-propylpentyl)aniline is prepared
following the method used in Example 31.
[1241] Step 1: Hydrogenation of nitrobenzene 40 and
2-propylpentanal gives
2-(3-(3-(2-propylpentylamino)phenyl)propyl)isoindoline-1,3-dione.
[1242] Step 2: Deprotection of
2-(3-(3-(2-propylpentylamino)phenyl)propyl)isoindoline-1,3-dione
gives Example 32.
Example 33
Preparation of 3-(3-aminopropyl)-N-(2-ethylbutyl)aniline
##STR00333##
[1244] 3-(3-Aminopropyl)-N-(2-ethylbutyl)aniline was prepared
following the method below.
[1245] Step 1: Trifluoroacetic anhydride (38.58 g, 0.18 mol) was
added dropwise over 10 min to a stirred solution of n-allylamine
(10.0 g, 0.17 mol) in CH.sub.2Cl.sub.2 at 0.degree. C. After
vigorous stirring at room temperature for 15 min, the reaction
mixture was quenched with saturated solution of NaHCO.sub.3 and
layers were separated. Aqueous layer was additionally extracted
with CH.sub.2Cl.sub.2. Combined organic layers were washed with
brine, dried over anhydrous NaSO.sub.4 and concentrated under
reduced pressure to give N-allyl-2,2,2-trifluoroacetamide as a
yellow liquid. Yield (17.5 g, 65%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.52 (bs, 1H), 5.88-5.79 (m, 1H), 5.29-5.23
(m, 2H), 3.97 (t, J=5.6 Hz, 2H).
[1246] Step 2: Palladium (II) acetate (0.449 g, 0.002 mol) was
added to a mixture of N-allyl-2,2,2-trifluoroacetamide (4.2 g, 0.02
mol), 1-bromo-3-nitrobenzene (5.09 g, 0.03 mol) and TBAA. The
reaction mixture was flushed with argon and heated under argon at
90.degree. C. for 4 h. The reaction mixture was partitioned between
EtOAc and water. Organic layer was dried over anhydrous
Na.sub.2SO.sub.4 and concentrated under reduced pressure to give
dark brown viscous liquid. Purification by flash chromatography (5%
to 30% EtOAc-hexane gradient) gave
2,2,2-trifluoro-N-(3-(3-nitrophenyl)allyl)acetamide as light yellow
solid. Yield (3.5 g, 61%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.77 (br.s, 1H), 8.27 (s, 1H), 8.10 (d, J=8.0 Hz, 1H), 7.93
(d, J=7.6 Hz, 1H), 7.63 (t, J=8.0 Hz, 1H), 6.71 (d, J=16.0 Hz, 1H),
6.51 (dt, J=5.6, 16.0 Hz, 1H), 4.02 (t, J=5.6 Hz, 2H).
[1247] Step 3: Tin(II) chloride dihydrate (3.28 g, 14.5 mmol) was
added to a solution of
(E)-2,2,2-trifluoro-N-(3-(3-nitrophenyl)allyl)acetamide (1.0 g,
3.64 mmol) in ethanol. The reaction mixture was stirred under
reflux overnight. The mixture was concentrated under reduced
pressure to give dark brown viscous liquid. The reaction mixture
was partitioned between sat NaHCO.sub.3 and EtOAc and then filtered
through Celite which was thoroughly washed with ethyl acetate.
Organic layer was separated and concentrated under reduced pressure
to give (E)-N-(3-(3-aminophenyl)allyl)-2,2,2-trifluoroacetamide as
a brown liquid. Yield (0.8 g, 89%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.70 (bs, 1H), 6.96 (t, J=7.8 Hz, 1H),
6.51-6.60 (m, 2H), 6.45-6.48 (m, 1H), 6.39 (d, J=16.0 Hz, 1H),
6.05-6.10 (m, 1H), 5.07 (bs, 2H), 3.95 (t, J=5.6 Hz, 2H); ESI MS
m/z 243.09 [M-H].sup.+.
[1248] Step 4: Hydrogenation of 2-ethylbutanal and
(E)-N-(3-(3-aminophenyl)allyl)-2,2,2-trifluoroacetamide following
the method used in Example 31 gave
N-(3-(3-(2-ethylbutylamino)phenyl)propyl)-2,2,2-trifluoroacetamide
as a colorless oil. Yield (0.5 g, 90%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.42 (bs, 1H), 6.94 (t, J=7.2 Hz, 1H),
6.38-6.37 (m, 2H), 6.32 (d, J=7.2 Hz, 1H), 5.40 (t, J=5.6 Hz, 1H),
3.21-3.16 (m, 2H), 2.86 (t, J=6.0 Hz, 2H), 2.43 (t, J=7.6 Hz, 2H),
1.77-1.70 (m, 2H), 1.48-1.41 (m, 1H), 1.39-1.31 (m, 4H), 0.85 (t,
J=7.6 Hz, 6H).
[1249] Step 5: A mixture of
N-(3-(3-(2-ethylbutylamino)phenyl)propyl)-2,2,2-trifluoroacetamide
(0.500 g, 1.51 mmol) and K.sub.2CO.sub.3 (0.631 g, 4.53 mmol) in
MeOH:water (2:1) was stirred at room temperature for 5 hr and
concentrated under reduced pressure. Purification by flash column
chromatography (5% to 20% of MeOH-DCM gradient) gave Example 33 as
a yellow oil. Yield (0.28 g, 76%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.93 (t, J=7.6 Hz, 1H), 6.38-6.36 (m, 2H),
6.31 (d, J=7.6 Hz, 1H), 5.39 (t, J=5.6 Hz, 1H), 3.46 (bs, 2H), 2.86
(t, J=6.0 Hz, 2H), 2.58 (t, J=7.2 Hz, 2H), 2.43 (t, J=8.0 Hz, 2H),
1.67-1.60 (m, 2H), 1.49-1.43 (m, 1H), 1.41-1.36 (m, 2H), 1.35-1.26
(m, 2H), 0.86 (t, J=7.6 Hz, 6H); .sup.13C NMR (100 MHz,
DMSO-d.sub.6) .delta.: 149.3, 142.3, 128.7, 115.4, 111.8, 109.3,
45.7, 40.5, 38.9, 33.4, 32.8, 23.3, 10.7; RP-HPLC (Method 3)
t.sub.R=3.71 min, 96.07% (AUC); ESI MS m/z 235.27 [M+H].sup.+.
Example 34
Preparation of 3-(3-aminopropyl)-N-benzylaniline
##STR00334##
[1251] 3-(3-Aminopropyl)-N-benzylaniline is prepared following the
method used in Example 31.
[1252] Step 1: Hydrogenation of nitrobenzene 40 and benzaldehyde
gives 2-(3-(3-(benzylamino)phenyl)propyl)isoindoline-1,3-dione.
[1253] Step 2: Deprotection of
2-(3-(3-(benzylamino)phenyl)propyl)isoindoline-1,3-dione gives
Example 34.
Example 35
Preparation of
3-amino-1-(3-(2-ethylbutylamino)phenyl)propan-1-ol
##STR00335##
[1255] 3-Amino-1-(3-(2-ethylbutylamino)phenyl)propan-1-ol was
prepared following the method used in Example 11.
[1256] Step 1: Hydrogenation of nitrobenzene 11 and 2-ethylbutanal
gave 3-(3-(2-ethylbutylamino)phenyl)-3-hydroxypropanenitrile as a
colorless oil. Yield (0.50 g, 78%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.01 (t, J=7.6 Hz, 1H), 6.61 (s, 1H), 6.50
(d, J=7.6 Hz, 1H), 6.46 (d, J=8.0 Hz, 1H), 5.77 (d, J=4.0 Hz, 1H),
5.55 (t, J=5.6 Hz, 1H), 4.72 (dd, J=4.8, 11.2 Hz, 1H), 2.88 (t,
J=6.0 Hz, 2H), 2.81 (dd, J=4.8, 16.4, 1H), 2.72 (dd, J=4.8, 16.4,
1H), 1.51-1.43 (m, 1H), 1.41-1.28 (m, 4H), 0.86 (t, J=7.6 Hz,
6H).
[1257] Step 2: BH.sub.3-Me.sub.2S reduction of
3-(3-(2-ethylbutylamino)phenyl)-3-hydroxypropanenitrile gave
Example 35 hydrochloride as a pale yellows semi-solid. Yield (0.28
g, 76%); .sup.1H NMR (400 MHz, DMSO-d.sub.6+5% D.sub.2O) .delta.
7.29 (t, J=7.6 Hz, 1H), 7.08 (s, 1H), 6.99 (m, 2H), 4.64 (dd,
J=4.0, 8.0 Hz, 1H), 3.04 (d, J=6.4 Hz, 2H), 2.91-2.80 (m, 2H),
1.89-1.75 (m, 2H), 1.56-1.49 (m, 1H), 1.44-1.27 (m, 4H), 0.82 (t,
J=7.6 Hz, 6H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 147.4,
138.4, 129.9, 124.43, 120.2, 118.7, 69.4, 53.4, 37.5, 36.4, 36.3,
22.8, 10.4; RP-HPLC (Method 6) t.sub.R=4.94 min, 96.74% (AUC); ESI
MS m/z 251.25 [M+H].sup.+.
Example 36
Preparation of
3-amino-1-(3-(2-ethylbutylamino)phenyl)propan-1-one
##STR00336##
[1259] 3-Amino-1-(3-(2-ethylbutylamino)phenyl)propan-1-one was
prepared following the method used in Examples 11 and 12.
[1260] Step 1: Protection of Example 35 with Boc.sub.2O following
the method used in Example 11 gave tert-butyl
3-(3-(2-ethylbutylamino)phenyl)-3-hydroxypropylcarbamate as a
colorless oil. Yield (0.55 g, 90%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.96 (t, J=8.0 Hz, 1H), 6.74 (br.s, 1H), 6.54
(s, 1H), 6.43 (d, J=7.6 Hz, 1H), 6.40 (d, J=8.0 Hz, 1H), 5.44 (t,
J=5.6 Hz, 1H), 5.01 (d, J=4.0 Hz, 1H), 4.37 (m, 1H), 2.98-2.92 (m,
2H), 2.87 (t, J=6.0 Hz, 2H), 1.66-1.61 (m, 2H), 1.51-1.43 (m, 1H),
1.36 (s, 9H), 1.30-1.23 (m, 4H), 0.86 (t, J=7.6 Hz, 6H).
[1261] Step 2: Oxidation of tert-butyl
3-(3-(2-ethylbutylamino)phenyl)-3-hydroxypropylcarbamate by
MnO.sub.2 following the method used in Example 12 gave tert-butyl
3-(3-(2-ethylbutylamino)phenyl)-3-oxopropylcarbamate as a pale
yellow oil. Yield (0.350 g, 86%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6+5% D.sub.2O) .delta. 7.17 (t, J=7.6 Hz, 1H), 7.06 (d,
J=7.6 Hz, 1H), 7.04 (s, 1H), 6.79 (d, J=7.6 Hz, 1H), 3.20 (t, J=6.4
Hz, 2H), 3.03 (t, J=6.8 Hz, 2H), 2.90 (d, J=6.0 Hz, 2H), 1.50-1.44
(m, 1H), 1.31 (s, 9H), 1.28-1.19 (m, 4H), 0.82 (t, J=7.6 Hz,
6H).
[1262] Step 3: Deprotection of tert-butyl
3-(3-(2-ethylbutylamino)phenyl)-3-oxopropylcarbamate following the
method used in Example 12 gave Example 36 hydrochloride as a yellow
oil. Yield (0.22 g, 90%); .sup.1H NMR (400 MHz, DMSO-d.sub.6+5%
D.sub.2O) .delta. 7.22 (t, J=7.6 Hz, 1H), 7.12-7.09 (m, 2H), 6.85
(d, J=7.6 Hz, 1H), 3.29 (t, J=6.4 Hz, 2H), 3.11 (t, J=6.0 Hz, 2H),
2.91 (d, J=6.0 Hz, 2H), 1.49-1.41 (m, 1H), 1.39-1.20 (m, 4H), 0.84
(t, J=7.2 Hz, 6H). RP-HPLC (Method 3) t.sub.R=4.49 min, 99.38%
(AUC); ESI MS m/z 249.22 [M+H].sup.+.
Example 37
Preparation of
3-amino-1-(3-(2-propylpentylamino)phenyl)propan-1-ol
##STR00337##
[1264] 3-Amino-1-(3-(2-propylpentylamino)phenyl)propan-1-ol was
prepared following the method described below.
[1265] Step 1: To a stirred solution of aniline 12 (1.0 g, 6.1
mmol) in EtOH:H.sub.2O (9:1) was added 2-propylpentyl
4-methylbenzenesulfonate (0.87 g, 3.08 mmol). The reaction mixture
was heated under reflux for 4 days, concentrated under reduced
pressure. The residue was diluted with water and extracted with
EtOAc three times. Combined organic layers were dried over
anhydrous sodium sulfate and concentrated under reduced pressure to
dryness. Purification by flash chromatography (25% EtOAc-hexanes)
gave 3-hydroxy-3-(3-(2-propylpentylamino)phenyl)propanenitrile as a
colorless semi-solid. Yield (0.3 g, 18%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.17 (t, J=7.6 Hz, 1H), 6.66 (d, J7.6 Hz, 1H),
6.62 (d, J=2.0 Hz, 1H), 6.57 (dd, J=2.0, 8.0 Hz, 1H), 4.95 (dt,
J=3.2, 6.0 Hz, 1H), 3.72 (bs, 1H), 3.02 (d, J=6.0 Hz, 2H), 2.80 (d,
J=6.8 Hz, 2H), 2.89 (d, J=3.2 Hz, 1H), 1.64-1.57 (m, 1H), 1.37-1.20
(m, 8H), 0.84 (t, J=6.8 Hz, 6H).
[1266] Step 2: BH.sub.3-Me.sub.2S reduction of
3-hydroxy-3-(3-(2-propylpentylamino)phenyl)propanenitrile following
the method used in Example 11 gave Example 37. Yield (0.19 g, 62%);
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 6.96 (t, J=7.6 Hz, 1H),
6.55 (s, 1H), 6.42 (d, J=7.2 Hz, 1H), 6.39 (d, J=8.0 Hz, 1H), 5.42
(t, J=5.6 Hz, 1H), 4.50 (t, J=6.0 Hz, 1H), 2.843 (d, J=6.4 Hz, 2H),
2.57 (t, J=6.4 Hz, 2H), 1.78-1.60 (m, 3H), 1.39-1.15 (m, 8H), 0.831
(t, J=6.4 Hz, 6H); RP-HPLC (Method 5) t.sub.R=5.67 min, 96.05%
(AUC); ESI MS m/z 279.27 [M+H].sup.+.
Example 38
Preparation of
3-amino-1-(3-(2-propylpentylamino)phenyl)propan-1-one
##STR00338##
[1268] 3-Amino-1-(3-(2-propylpentylamino)phenyl)propan-1-one was
prepared following the method used in Examples 11 and 12.
[1269] Step 1: Protection of Example 37 with Boc.sub.2O following
the method used in Example 11 gave tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)-1-hydroxypropyl)phenyl)(2-propylpentyl-
)carbamate as a pale yellow semi-solid. Yield (0.6 g, 46%); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.30 (t, J=8.0 Hz, 1H), 7.19-7.16
(m, 2H), 7.09 (d, J=7.2 Hz, 1H), 4.90 (bs, 1H), 4.73-4.64 (m, 1H),
3.55 (dd, J=7.2, 14.4 Hz, 2H), 3.22-3.14 (m, 2H), 3.01 (d, J=6.0
Hz, 1H), 1.87-1.81 (m, 2H), 1.45 (s, 9H), 1.43 (s, 9H), 1.33 (m,
5H), 1.21 (m, 4H), 0.89 (m, 3H), 0.81 (m, 3H).
[1270] Step 2: Oxidation often-butyl
(3-(3-((tert-butoxycarbonyl)amino)-1-hydroxypropyl)phenyl)(2-propylpentyl-
)carbamate by Des-Martin periodinane following the method used in
Example 40 gave tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(2-propylpentyl)carbam-
ate as a pale yellow semi-solid. Yield (0.25 g, 55%); .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.79-7.75 (m, 2H), 7.43 (d, J=4.8 Hz,
2H), 5.13 (bs, 1H), 3.61 (d, J=7.6 Hz, 2H), 3.55-3.52 (m, 2H), 3.18
(t, J=5.6 Hz, 2H), 1.56 (s, 18H), 1.44-1.21 (m, 9H), 0.81 (t, J=6.0
Hz, 6H).
[1271] Step 3: Deprotection of tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(2-propylpentyl)carbam-
ate following the method used in Example 12 gave Example 38
hydrochloride as a white solid. Yield (0.03 g, 46%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.85 (m, 3H), 7.24 (t, J=7.6 Hz,
1H), 7.14-7.12 (m, 2H), 6.89 (d, J=7.6 Hz, 1H), 3.32 (t, J=6.4 Hz,
2H), 3.14-3.09 (m, 2H), 2.94 (d, J=6.0, 2H), 1.62 (bs, 1H),
1.34-1.23 (m, 9H), 0.87 (t, J=6.4 Hz, 6H); RP-HPLC (Method 6)
t.sub.R=5.98 min, 79.55% (AUC); ESI MS m/z 277.29 [M+H].sup.+.
Example 39
Preparation of
3-amino-1-(3-(cyclopentylmethylamino)phenyl)propan-1-ol
##STR00339##
[1273] 3-Amino-1-(3-(cyclopentylmethylamino)phenyl)propan-1-ol was
prepared following the method used in Example 35.
[1274] Step 1: Hydrogenation of nitrobenzene 11 and
cyclopentylcarbaldehyde gave
3-(3-(cyclopentylmethylamino)phenyl)-3-hydroxypropanenitrile as a
brown oil. Yield (2.42 g, 95%); .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.17 (t, J=7.6 Hz, 1H), 6.66 (d, J=7.6 Hz, 1H), 6.62 (s,
1H), 6.57 (dd, J=2.0, 8.0 Hz, 1H), 4.94 (t, J=6.0 Hz, 1H), 3.72
(bs, 1H), 3.02 (d, J=7.2 Hz, 2H), 2.76-2.73 (m, 2H), 2.18-2.11 (m,
1H), 1.86-1.79 (m, 2H), 1.67-1.50 (m, 4H), 1.30-1.22 (m, 2H).
[1275] Step 2: BH.sub.3-Me.sub.2S reduction of
3-(3-(cyclopentylmethylamino)phenyl)-3-hydroxypropanenitrile gave,
after purification following the method used in Example 11, Example
39 hydrochloride as a pale yellow semi-solid. Yield (2.0 g, 81%);
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.54 (t, J=7.6 Hz, 2H),
7.48 (d, J=8.0 Hz, 1H), 7.37 (d, J=7.6 Hz, 1H), 4.90 (m, 1H), 3.37
(d, J=7.6 Hz, 2H), 3.14-3.09 (m, 2H), 2.25-2.21 (m, 1H), 2.07-1.98
(m, 2H), 1.96-1.87 (m, 2H), 1.75-1.67 (m, 2H), 1.65-1.62 (m, 2H),
1.37-1.30 (m, 2H); RP-HPLC (Method 6) t.sub.R=4.75 min, 97.99%
(AUC); ESI MS m/z 249.30 [M+H].sup.+.
Example 40
Preparation of
3-amino-1-(3-(cyclopentylmethylamino)phenyl)propan-1-one
##STR00340##
[1277] 3-Amino-1-(3-(cyclopentylmethylamino)phenyl)propan-1-one was
prepared following the method used in Examples 11 and 12.
[1278] Step 1: Protection of Example 39 hydrochloride following the
method used in Example 11 gave a mixture of tert-butyl
(3-(3-((cyclopentylmethyl)amino)phenyl)-3-hydroxypropyl)carbamate
and tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)-1-hydroxypropyl)phenyl)(cyclopentylmet-
hyl)carbamate as a pale yellow oil. Yield (2.0 g, 71%); .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.31 (t, J=8.0 Hz, 1H), 7.18 (m, 2H),
7.08 (d, J=6.8 Hz, 1H), 4.91 (s, 1H), 4.73 (s, 1H), 3.59 (d, J=7.6
Hz, 2H), 3.51 (d, J=5.2 Hz, 1H), 3.33 (s, 1H), 3.16 (dd, J=5.2,
14.4 Hz, 1H), 2.04-1.97 (m, 1H), 1.83 (bs, 2H), 1.59 (s, 2H), 1.45
(bs, 11H), 1.42 (s, 9H), 1.25-1.18 (m, 4H).
[1279] Step 2: To a stirred solution of the above mixture (0.6 g,
1.72 mmol) in CH.sub.2Cl.sub.2 was added Des-Martin periodinane
(0.80 g, 1.89 mmol). After stirring at room temperature for 1 h,
the reaction mixture was concentrated under reduced pressure.
Purification by column chromatography (5% to 20% EtOAc-hexanes)
gave a mixture of tert-butyl
(3-(3-((cyclopentylmethyl)amino)phenyl)-3-oxopropyl)carbamate and
tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(cyclopentylmethyl)car-
bamate as a pale yellow oil. Yield (0.55 g, 92%); .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.78-7.76 (m, 2H), 7.42 (d, J=4.8 Hz, 2H),
5.14 (s, 1H), 3.63 (d, J=7.6 Hz, 2H), 3.56-3.52 (m, 2H), 3.19 (t,
J=5.2 Hz, 2H), 2.03-1.95 (m, 1H), 1.64-1.58 (m, 4H), 1.55-1.48 (m,
2H), 1.42 (s, 18H), 1.23-1.16 (m, 2H).
[1280] Step 3: Deprotection of the above mixture following the
method used in Example 12 gave Example 40 hydrochloride as a white
solid. Yield (0.17 g, 95%); .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.13 (s, 2H), 7.76-7.74 (m, 2H), 3.53 (t, J=6.0 Hz, 2H),
3.43 (d, J=7.2 Hz, 2H), 3.37 (t, J=6.0 Hz, 2H), 2.31-2.23 (quintet,
J=7.6 Hz, 1H), 1.95-1.89 (m, 2H), 1.78-1.72 (m, 2H), 1.70-1.60 (m,
2H), 1.40-1.31 (m, 2H); .sup.13C NMR (100 MHz, CD.sub.3OD) .delta.
197.3, 139.2, 138.1, 132.2, 130.2, 129.0, 123.3, 58.5, 38.2, 36.8,
35.7, 31.6, 26.1; RP-HPLC (Method 6) t.sub.R=5.03 min, 95.24%
(AUC); ESI MS m/z 247.24 [M+H].sup.+.
Example 41
Preparation of
3-amino-1-(3-(5-(benzyloxy)pentylamino)phenyl)propan-1-ol
##STR00341##
[1282] 3-Amino-1-(3-(5-(benzyloxy)pentylamino)phenyl)propan-1-olwas
prepared following the method used in Example 11.
[1283] Step 1: Hydrogenation of nitrobenzene 11 and
5-(benzyloxy)pentanal gave
34345-(benzyloxy)pentylamino)phenyl)-3-hydroxypropanenitrile as a
colorless oil. Yield (0.90 g, 66%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.36-7.25 (m, 5H), 7.01 (t, J=7.6 Hz, 1H),
6.59 (s, 1H), 6.52 (d, J=7.6 Hz, 1H), 6.44 (d, J=7.6 Hz, 1H), 5.77
(d, J=4.4 Hz, 1H), 5.56 (t, J=5.2 Hz, 1H), 4.74-4.70 (m, 1H), 4.44
(s, 2H), 3.42 (t, J=6.8 Hz, 2H), 3.00-2.95 (m, 2H), 2.80 (dd,
J=4.8, 16.4 Hz, 1H), 2.73 (dd, J=4.8, 16.4 Hz, 1H), 1.60-1.51 (m,
4H), 1.44-1.20 (m, 2H).
[1284] Step 2: BH.sub.3-Me.sub.2S reduction of
3-(3-(5-(benzyloxy)pentylamino)phenyl)-3-hydroxypropanenitrile gave
Example 41 hydrochloride as a white solid. Yield (0.18 g, 66%).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.95 (bs, 3H),
7.36-7.20 (m, 9H), 4.70-4.69 (m, 1H), 4.44 (s, 2H), 3.41 (t, J=6.4
Hz, 2H), 3.19-3.14 (m, 2H), 2.86-2.85 (m, 2H), 1.90-1.80 (m, 2H),
1.67-1.52 (m, 4H), 1.43-1.23 (m, 2H); .sup.13CNMR (400 MHz,
DMSO-d.sub.6) .delta. 147.6, 138.9, 130.2, 130.1, 128.7, 127.9,
127.8, 125.2, 120.7, 119.1, 72.3, 69.7, 50.4, 36.7, 36.6, 36.4,
29.0, 25.8, 23.1; RP-HPLC Method 5) t.sub.R=5.30 min, 94.93% (AUC);
ESI MS m/z 343.30 [M+H].sup.+.
Example 42
Preparation of
3-amino-1-(3-(5-(benzyloxy)pentylamino)phenyl)propan-1-one
##STR00342##
[1286] 3-Amino-1-(3-(5-(benzyloxy)pentylamino)phenyl)propan-1-one
was prepared following the method used in Example 38.
[1287] Step 1: Protection of Example 41 with Boc.sub.2O gave a
mixture of tert-butyl
(3-(3-((5-(benzyloxy)pentyl)amino)phenyl)-3-hydroxypropyl)carbamate
(minor component) and tert-butyl
(5-(benzyloxy)pentyl)(3-(3-((tert-butoxycarbonyl)amino)-1-hydroxypropyl)p-
henyl)carbamate (major component) as a colorless oil. Yield (2.0 g,
98%); Major .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.36-7.29 (m,
4H), 7.23 (m, 1H), 7.18 (d, J=4.4 Hz, 1H), 7.14-7.12 (m, 2H), 6.64
(d, J=7.2 Hz, 1H), 4.90 (bs, 1H), 4.65 (bs, 1H), 4.50 (s, 2H),
3.62-3.60 (m, 2H), 3.48 (t, J=6.4 Hz, 2H), 3.14-3.10 (m, 2H),
1.85-1.83 (m, 2H), 1.68-1.57 (m, 6H), 1.46 (s, 9H), 1.45 (s, 9H).
Minor .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.28-7.24 (m, 5H),
7.18-7.16 (m, 1H), 7.07-7.05 (m, 2H), 6.60 (bs, 1H), 6.50 (bs, 1H),
4.70 (bs, 1H), 4.47 (s, 2H), 3.59 (m, 2H), 3.43 (t, J=6.4 Hz, 2H),
3.17-3.14 (m, 2H), 2.90 (bs, 1H), 1.83-1.81 (m, 2H), 1.68-1.57 (m,
6H), 1.41 (s, 9H).
[1288] Step 2: Oxidation of the above mixture by Des-Martin
periodinane following the method used in Example 40 gave a mixture
of tert-butyl
(3-(3-((5-(benzyloxy)pentyl)amino)phenyl)-3-oxopropyl)carbamate
(minor component) and tert-butyl
(5-(benzyloxy)pentyl)(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)c-
arbamate as a colorless oil. Yield (0.3 g, 25%); .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.78-7.75 (m, 2H), 7.42 (d, J=5.6 Hz, 2H),
7.34-7.31 (m, 5H), 5.12 (bs, 1H), 4.47 (s, 2H), 3.65 (t, J=7.6 Hz,
2H), 3.54-3.49 (m, 2H), 3.43 (t, J=6.4 Hz, 2H), 3.17 (bs, 2H),
1.62-1.55 (m, 4H), 1.43 (s, 18H), 1.37-1.35 (m, 2H).
[1289] Step 3: Deprotection of the above mixture gave Example 42
hydrochloride as an off-white solid. Yield (0.2 g, 76%); .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 8.11 (d, J=6.8 Hz, 1H), 8.07 (s,
1H), 7.75-7.69 (m, 2H), 7.35-7.31 (m, 4H), 7.29-7.24 (m, 1H), 4.49
(s, 2H), 3.53-3.49 (m, 4H), 3.42 (t, J=7.6 Hz, 2H), 3.36-3.31 (m,
2H), 1.82-1.70 (m, 2H), 1.68-1.63 (m, 2H), 1.57-1.50 (m, 2H);
RP-HPLC (Method 3) t.sub.R=4.54 min, 90.10% (AUC); ESI MS m/z
341.31 [M+H].sup.+.
Example 43
Preparation of
5-(3-(3-amino-1-hydroxypropyl)phenylamino)pentan-1-ol
##STR00343##
[1291] 5-(3-(3-Amino-1-hydroxypropyl)phenylamino)pentan-1-ol is
prepared following the method described below.
[1292] Step 1: A mixture of Example 41 and Pd(OH).sub.2/C (20% wt)
in absolute EtOH is stirred at room temperature under hydrogen
atmosphere until no starting material is seen by TLC. The reaction
mixture is filtered through Celite and concentrated under reduced
pressure to give Example 43.
Example 44
Preparation of
3-amino-1-(3-(5-hydroxypentylamino)phenyl)propan-1-one
##STR00344##
[1294] 3-Amino-1-(3-(5-hydroxypentylamino)phenyl)propan-1-one is
prepared following the method described below.
[1295] Step 1: Protection of Example 43 with Boc.sub.2O following
the method used in Example 11 gives tert-butyl
3-hydroxy-3-(3-(5-hydroxypentylamino)phenyl)propylcarbamate.
[1296] Step 2: MnO.sub.2 oxidation of tert-butyl
3-hydroxy-3-(3-(5-hydroxypentylamino)phenyl)propylcarbamate
following the method used in Example 12 gives tert-butyl
3-(3-(5-hydroxypentylamino)phenyl)-3-oxopropylcarbamate.
[1297] Step 3: Deprotection of tert-butyl
3-(3-(5-hydroxypentylamino)phenyl)-3-oxopropylcarbamate following
the method used in Example 12 gives Example 44 hydrochloride.
Example 45
Preparation of
3-amino-1-(3-(5-methoxypentylamino)phenyl)propan-1-ol
##STR00345##
[1299] 3-Amino-1-(3-(5-methoxypentylamino)phenyl)propan-1-ol was
prepared following the method used in Example 37.
[1300] Step 1: A mixture of 5-methoxypentanal (0.644 g, 5.54 mmol),
3-(3-aminophenyl)-3-hydroxypropanenitrile (12) (1.0 g, 6.16 mmol)
and activated molecular sieves in methanol was stirred at RT for 8
hrs. NaBH.sub.4 (0.937 g, 24.6 mmol) was added portion wise to the
reaction mixture at 0.degree. C. The reaction mixture was stirred
at RT overnight. The reaction mixture was filtered through Celite
and the filtrate was concentrated under reduced pressure.
Purification by column chromatography (100-200 silica, 0% to 70%
EtOAc-hexanes gradient) gave
3-hydroxy-3-(3-(5-methoxypentylamino)phenyl)propanenitrile as a
yellow oil. Yield (0.34 g, 21%); .sup.1H NMR (400 MHz, CDCl3)
.delta. 7.17 (t, J=8.0 Hz, 1H), 6.66 (d, J=7.6 Hz, 1H), 6.62 (s,
1H), 6.56 (d, J=8.0 Hz, 1H), 4.95 (t, J=6.4 Hz, 1H), 3.39 (t, J=6.4
Hz, 2H), 3.33 (s, 3H), 3.12 (t, J=7.2 Hz, 2H), 2.75 (d, J=6.4 Hz,
2H), 1.68-1.59 (m, 4H), 1.51-1.43 (m, 2H).
[1301] Step 2: BH.sub.3-Me.sub.2S reduction of
3-hydroxy-3-(3-(5-methoxypentylamino)phenyl)propanenitrile
following the method used in Example 11 gave Example 45
hydrochloride as a colorless oil. Yield (0.25 g, 72%); .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 7.60-7.53 (m, 3H), 7.40 (d, J=7.6 Hz,
1H), 4.94-4.92 (m, 1H), 3.42-3.38 (m, 4H), 3.31 (s, 3H), 3.17-3.10
(m, 2H), 2.07-2.03 (m, 1H), 2.01-1.94 (m, 1H), 1.81-1.73 (m, 2H),
1.63-1.60 (m, 2H), 1.58-1.49 (m, 2H); RP-HPLC (Method 6)
t.sub.R=4.02 min, 82.18% (AUC); ESI MS m/z 267.28 [M+H].sup.+.
Example 46
Preparation of
3-amino-1-(3-(5-methoxypentylamino)phenyl)propan-1-one
##STR00346##
[1303] 3-Amino-1-(3-(5-methoxypentylamino)phenyl)propan-1-one was
prepared following the method used in Example 38.
[1304] Step 1: Protection of Example 45 with Boc.sub.2O following
the method used in Example 11 gave tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(5-methoxypentyl)carba-
mate as a colorless oil. Yield (0.22 g); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.29 (t, J=7.2 Hz, 1H), 7.19-7.17 (m, 2H), 7.08
(d, J=8.0, 1H), 4.92-4.90 (m, 1H), 4.74-4.71 (m, 1H), 3.59 (t,
J=8.0, 2H), 3.57 (bs, 1H), 3.38-3.32 (m, 2H), 3.31 (s, 3H),
3.20-3.12 (m, 2H), 1.85-1.83 (m, 2H), 1.56 (s, 9H), 1.45 (m, 4H),
1.47 (s, 9H), 1.34-1.30 (m, 2H).
[1305] Step 2: Oxidation of tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(5-methoxypentyl)carba-
mate by Des-Martin periodinane following the method used in Example
40 gave tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(5-methoxypentyl)carba-
mate as a colorless oil. Yield (0.12 g, 53%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.78-7.76 (m, 2H), 7.43-7.42 (m, 2H), 5.13 (bs,
1H), 3.65 (t, J=7.6 Hz, 2H), 3.55-3.52 (m, 2H), 3.35-3.32 (t, J=6.4
Hz, 2H), 3.30 (s, 3H), 3.20-3.17 (t, J=5.6 Hz, 2H), 1.66-1.52 (m,
4H), 1.43 (s, 9H), 1.42 (s, 9H), 1.39-1.30 (m, 2H).
[1306] Step 3: Deprotection of tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(5-methoxypentyl)carba-
mate following the method used in Example 12 gave Example 46
hydrochloride as a yellow solid. Yield (0.07 g, 70%); .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 7.95 (d, J=7.2 Hz, 1H), 7.91 (s, 1H),
7.64 (t, J=8.0 Hz, 1H), 7.56 (d, J=6.4 Hz, 1H), 3.49 (t, J=6.0 Hz,
2H), 3.41-3.34 (m, 9H), 1.79-1.71 (m, 2H), 1.66-1.59 (m, 2H),
1.53-1.46 (m, 2H); (RP-HPLC Method 6) t.sub.R=4.42 min, 96.0%
(AUC); ESI MS m/z 265.26 [M+H].sup.+.
Example 47
Preparation of
3-amino-1-(3-((2-methoxybenzyl)amino)phenyl)propan-1-ol
##STR00347##
[1308] 3-Amino-1-(3-((2-methoxybenzyl)amino)phenyl)propan-1-ol was
prepared following the method used in Example 11.
[1309] Step 1: Hydrogenation of nitrobenzene 11 and
2-methoxybenzaldehyde gave
3-hydroxy-3-(3-(2-methoxybenzylamino)phenyl)propanenitrile as a
yellow oil. Yield (1.4 g, 95%); .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.29-7.27 (m, 1H), 7.16 (t, J=8.0 Hz, 1H), 6.94 (t, J=7.2
Hz, 1H), 6.90 (d, J=9.2 Hz, 2H), 6.68-6.67 (m, 2H), 6.61 (d, J=8.0
Hz, 1H), 4.93 (br.s, 1H), 4.69 (d, J=6.8 Hz, 1H), 4.33 (s, 2H),
3.87 (s, 3H), 2.78-2.73 (m, 2H).
[1310] Step 2: BH.sub.3-Me.sub.2S reduction of
3-hydroxy-3-(3-(2-methoxybenzylamino)phenyl)propanenitrile gave
crude 3-amino-1-(3-(2-methoxybenzylamino)phenyl)propan-1-ol
hydrochloride as a pale yellow oil. Yield (1.22 g, 86%); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.30 (dd, J=1.2, 6.4 Hz, 1H),
7.23 (dd, J=1.2, 7.6 Hz, 1H), 7.11 (t, J=8.0 Hz, 1H), 6.91-6.87 (m,
2H), 6.72 (bs, 1H), 6.66 (d, J=7.2 Hz, 1H), 6.53 (dd, J=2.0, 8.0
Hz, 1H), 4.86-4.83 (m, 1H), 4.33 (s, 2H), 4.15 (bs, 1H), 3.85 (s,
3H), 3.06-3.01 (m, 1H), 2.94-2.90 (m, 1H), 1.88-1.76 (m, 2H).
[1311] Step 3: Step 3: Boc protection of
3-amino-1-(3-(2-methoxybenzylamino)phenyl)propan-1-ol hydrochloride
gave a mixture of mono- and di-Boc products which was used directly
in the next step without further purification. Major component:
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.30 (d, J=7.6 Hz, 1H),
7.23-7.18 (m, 4H), 6.92-6.87 (m, 2H), 6.66 (d, J=7.6 Hz, 1H), 4.85
(s, 2H), 4.69-4.63 (m, 1H), 3.72 (s, 3H), 3.18-3.10 (m, 2H),
1.83-1.77 (m, 2H), 1.45 (s, 9H), 1.41 (s, 9H). Minor component:
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.28 (m, 1H), 7.14-7.07
(m, 4H), 6.82 (d, J=8.4 Hz, 2H), 6.68 (bs, 1H), 6.56 (d, J=9.6 Hz,
1H), 4.86-4.85 (m, 1H), 4.32 (s, 2H), 3.86 (s, 3H), 3.45-3.43 (m,
2H), 1.86-1.83 (m, 2H), 1.45 (s, 9H).
[1312] Step 4: Deprotection of the above mixture following the
method used in Example 11 gave Example 47 hydrochloride as a yellow
solid. Yield (0.194 g, 66%); .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.53-7.46 (m, 3H), 7.44-7.42 (m, 1H), 7.28-7.25 (m, 2H),
7.10 (d, J=8.0 Hz, 1H), 6.95 (t, J=7.2 Hz, 1H), 4.86 (m, 1H), 4.56
(s, 2H), 3.94 (s, 3H), 3.12-3.06 (m, 2H), 2.03-1.99 (m, 1H),
1.95-1.87 (m, 1H);.sup.13C NMR (100 MHz, CD.sub.3OD) .delta. 158.1,
147.4, 135.2, 131.7, 131.5, 129.9, 126.6, 121.6, 120.5, 119.9,
118.4, 110.6, 70.6, 54.8, 51.7, 37.2, 35.7; RP-HPLC (Method 6)
t.sub.R=4.49 min, 96.74% (AUC); ESI MS m/z 287.23 [M+H].sup.+.
Example 48
Preparation of
3-amino-1-(3-(2-methoxybenzylamino)phenyl)propan-1-one
##STR00348##
[1314] 3-Amino-1-(3-(2-methoxybenzylamino)phenyl)propan-1-one was
prepared following the method used in Example 12.
[1315] Step 1: Protection of Example 47 with Boc.sub.2O gave a
mixture of tert-butyl
3-hydroxy-3-(3-(2-methoxybenzylamino)phenyl)propylcarbamate and
tert-butyl
tert-butylcarbonyl(3-hydroxy-3-(3-(2-methoxybenzylamino)phenyl)propyl)car-
bamate as a yellow oil. Yield (0.4 g, 63%); .sup.1H NMR (400 MHz,
CDCl3) .delta. 7.80 (bs, 1H), 7.70 (d, J=7.2 Hz, 1H), 7.40-7.36 (m,
1H), 7.34 (t, J=7.6 Hz, 1H), 7.26-7.19 (m, 2H), 6.90 (t, J=7.2 Hz,
1H), 6.82 (d, J=8.0, 1H), 5.11 (bs, 1H), 4.89 (s, 2H), 3.71 (s,
3H), 3.51 (m, 2H), 3.12 (m, 2H), 1.42 (s, 18H).
[1316] Step 2: Oxidation of the above mixture with Des-Martin
periodinane following the method used in Example 20 gave a mixture
of tert-butyl
3-oxo-3-(3-(2-methoxybenzylamino)phenyl)propylcarbamate and
tert-butyl
tert-butylcarbonyl(3-oxo-3-(3-(2-methoxybenzylamino)phenyl)propyl)carbama-
te which was directly used in the next step without further
purification.
[1317] Step 3: Deprotection of the above mixture following the
method used in Example 12 gave Example 48 hydrochloride as an
off-white solid. Yield (0.2 g, 59%); .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 8.10 (d, J=7.6 Hz, 1H), 7.99 (s, 1H), 7.69 (t,
J=8.0 Hz, 1H), 7.61 (d, J=8.0 Hz, 1H), 7.43 (dt, J=1.2, 8.0 Hz,
1H), 7.29 (d, J=7.2 Hz, 1H), 7.09 (d, J=8.4 Hz, 1H), 6.95 (t, J=7.6
Hz, 1H), 4.63 (s, 2H), 3.93 (s, 3H), 3.49 (t, J=6.4 Hz, 2H),
3.37-3.34 (m, 2H);.sup.13C NMR (100 MHz, DMSO-d.sub.6)
.delta.:197.2, 159.5, 138.9, 137.4, 133.1, 133.0, 131.9, 130.3,
129.0, 123.5, 121.9, 119.7, 112.0, 56.3, 52.7, 36.8, 35.7; RP-HPLC
(Method 6) t.sub.R=4.84 min, 99.31% (AUC); ESI MS m/z 285.3
[M+H].sup.+.
Example 49
Preparation of 3-amino-1-(3-(phenethylamino)phenyl)propan-1-ol
##STR00349##
[1319] 3-Amino-1-(3-(phenethylamino)phenyl)propan-1-ol was prepared
following the method described below.
[1320] Step 1: Hydrogenation of aniline 12 and 2-phenylacetaldehyde
gives 3-hydroxy-3-(3-(phenethylamino)phenyl)propanenitrile. A
mixture of aniline 12 (1.00 g, 6.17 mmol), 2-phenylacetaldehyde
(0.66 g, 5.55 mmol) and A-3 molecular sieves in MeOH was stirred
for 18 h and then NaBH.sub.4 (1.16 g, 30.8 mmol) was added and the
reaction mixture was stirred overnight. The reaction mixture was
filtered through Celite, concentrated under reduced pressure.
Purification by column chromatography (100-200 silica mesh, 20%
EtOAc-hexane) gave
3-hydroxy-3-(3-(phenethylamino)phenyl)propanenitrile as a yellow
liquid. Yield (0.432 g, 27%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.32-7.26 (m, 4H), 7.20 (t, J=6.4 Hz, 1H), 7.04 (t, J=7.6
Hz, 1H), 6.64 (s, 1H), 6.55 (d, J=7.2 Hz, 1H), 6.50 (d, J=8.4 Hz,
1H), 5.77 (d, J=4.4 Hz, 1H), 5.68 (t, J=5.6 Hz, 1H), 4.76-4.72 (m,
1H), 3.26-3.21 (m, 2H), 2.85-2.81 (m, 3H), 2.79-2.72 (m, 1H).
[1321] Step 2: BH.sub.3-Me.sub.2S reduction of
3-hydroxy-3-(3-(phenethylamino)phenyl)propanenitrile following the
method used in Example 11 gave Example 49 as a yellow semi-solid.
Yield (0.15 g, 38%); .sup.1H NMR (400 MHz, MeOD) .delta. 7.31 (t,
J=7.6 Hz, 2H), 7.27-7.20 (m, 4H), 6.94 (s, 1H), 6.91 (d, J=7.2 Hz,
1H), 6.82 (d, J=6.8 Hz, 1H), 4.81-4.77 (m, 1H), 3.44 (t, J=7.2 Hz,
2H), 3.10-3.02 (m, 2H), 2.93 (t, J=7.2 Hz, 2H), 1.96 (m, 2H);
RP-HPLC (Method 3) t.sub.R=3.42 min, 96.13% (AUC); ESI MS m/z
271.25 [M+H].sup.+.
Example 50
Preparation of 3-amino-1-(3-(phenethylamino)phenyl)propan-1-one
##STR00350##
[1323] 3-Amino-1-(3-(phenethylamino)phenyl)propan-1-one was
prepared following the method used in Example 38.
[1324] Step 1: Protection of Example 49 with Boc.sub.2O gave
tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(phenethyl)carbamate
as a yellow oil. Yield (1.2 g, 98%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) 7.29-7.23 (m, 3H), 7.21 (d, J=7.2 Hz, 1H), 7.16 (d,
J=7.2 Hz, 3H), 7.10 (s, 1H), 7.04 (d, J=7.6 Hz, 1H), 6.78 (m, 1H),
5.25 (d, J=4.4 Hz, 1H), 4.54 (m, 1H), 3.79 (t, J=7.2 Hz, 2H), 2.97
(t, J=6.0 Hz, 2H), 2.77-2.71 (m, 2H), 1.70-1.66 (m, 2H), 1.42 (s,
9H), 1.34 (s, 9H).
[1325] Step 2: Oxidation of tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(phenethyl)carbamate
by Des-Martin periodinane gave tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(phenethyl)carbamate
as a yellow oil. Yield (0.9 g, 76%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.75 (d, J=7.2 Hz, 1H), 7.66 (s, 1H),
7.50-7.43 (m, 2H), 7.27 (t, J=7.6 Hz, 2H), 7.21 (d, J=7.2 Hz, 1H),
7.16 (d, J=7.6 Hz, 2H), 6.85 (bs,1H), 3.86 (t, J=7.2 Hz, 2H), 3.26
(t, J=6.0 Hz, 2H), 3.11 (t, J=6.8 Hz, 2H), 2.78 (t, J=7.2 Hz, 2H),
1.35 (s, 18H).
[1326] Step 3: Deprotection of tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(phenethyl)carbamate
gave Example 50 hydrochloride as a white solid. Yield (0.5 g, 94%);
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.05 (m, 2H), 7.73-7.69
(m, 2H), 7.35-7.24 (m, 5H), 3.69-3.65 (m, 2H), 3.51 (t, J=6.4 Hz,
2H), 3.36 (t, J=6.0 Hz, 2H), 3.07 (t, J=8.0 Hz, 2H); RP-HPLC
(Method 6) t.sub.R=4.93 min, 93.74% (AUC); ESI MS m/z 269.28
[M+H].sup.+.
Example 51
Preparation of
3-amino-1-(3-(3-cyclohexylpropylamino)phenyl)propan-1-ol
##STR00351##
[1328] 3-Amino-1-(3-(3-cyclohexylpropylamino)phenyl)propan-1-ol was
prepared following the method used in Example 35.
[1329] Step 1: Hydrogenation of nitrobenzene 11 and
3-cyclohexylpropanal gave
3-(3-(3-cyclohexylpropylamino)phenyl)-3-hydroxypropanenitrile as a
colorless semi-solid. Yield (0.32 g, 71%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.17 (t, J=7.6 Hz, 1H), 6.67 (d, J=7.6 Hz, 1H),
6.62 (s, 1H), 6.57 (d, J=8.0, 1H), 4.95 (t, J=6.0, 1H), 3.74-3.70
(bs, 1H), 3.09 (t, J=7.2 Hz, 2H), 2.76 (d, J=6.0 Hz, 2H), 2.23
(bs,1H), 1.72-1.70 (m, 4H), 1.66-1.58 (m, 2H), 1.31-1.12 (m, 7H)
0.92-0.87 (m, 2H).
[1330] Step 2: BH.sub.3-Me.sub.2S reduction of
3-(3-(3-cyclohexylpropylamino)phenyl)-3-hydroxypropanenitrile gave
Example 51 hydrochloride as a colorless semi-solid. Yield (0.250 g,
82%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 6.97 (t, J=8.0
Hz, 1H), 6.52 (s, 1H), 6.46 (d, J=7.2 Hz, 1H), 6.38 (d, J=8.0 Hz,
1H), 5.43 (t, J=4.8 Hz, 1H), 4.54-4.48 (m, 1H), 2.96-2.91 (q, J=6.4
Hz, 2H), 2.67-2.60 (m, 2H), 1.70-1.61 (m, 6H), 1.54-1.51 (m, 2H),
1.25-1.06 (m, 6H), 0.90-0.84 (m, 2H); .sup.13C NMR (100 MHz,
DMSO-d.sub.6) .delta. 148.4, 146.5, 127.9, 112.6, 109.7, 108.9,
71.1, 42.8, 41.4, 38.37, 36.5, 34.1, 32.5, 25.8, 25.6, 25.4;
RP-HPLC (Method 3) t.sub.R=4.13 min, 92.02% (AUC); ESI MS m/z
291.30 [M+H].sup.+.
Example 52
Preparation of
3-amino-1-(3-(3-cyclohexylpropylamino)phenyl)propan-1-one
##STR00352##
[1332] 3-Amino-1-(3-(3-cyclohexylpropylamino)phenyl)propan-1-one
was prepared following the method used in Example 40.
[1333] Step 1: Protection of Example 51 hydrochloride following the
method used in Example 11 gave a mixture of tert-butyl
3-(3-(3-cyclohexylpropylamino)phenyl)-3-hydroxypropylcarbamate and
tert-butyl
tert-butoxycarbonyloxy(3-(3-(3-cyclohexylpropylamino)phenyl)-3-hydroxypro-
pyl)carbamate as a colorless semi-solid which was used in the next
step. Yield (1.2 g, 48%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.29 (t, J=7.6 Hz, 1H), 7.19 (s, 2H), 7.17 (d, J=8.0 Hz,
1H), 7.14 (t, J=7.6 Hz, 1H), 7.08 (d, J=8.0 Hz, 1H), 6.64-6.61 (m,
2H), 6.51 (d, J=7.6 Hz, 1H), 4.90 (bs, 2H), 4.73-4.65 (m, 2H), 3.57
(m, 3H), 3.39 (bs, 1H), 3.19-3.15 (m, 2H), 3.09 (t, J=7.2 Hz, 2H),
2.90 (bs, 1H), 1.85-1.71 (m, 2H), 1.69-1.61 (m, 12H), 1.48 (s, 9H),
1.45 (s, 6H), 1.42-1.20 (m, 3H), 1.17-1.12 (m, 5H) 0.89-0.81 (m,
3H).
[1334] Step 2: Oxidation of the above mixture by Des-Martin
periodinane following the method used in Example 20 gave a mixture
of tert-butyl
3-(3-(3-cyclohexylpropylamino)phenyl)-3-oxopropylcarbamate and
tert-butyl
tert-butoxycarbonyloxy(3-(3-(cyclohexylpropylamino)phenyl)-3-oxopropyl)ca-
rbamate as a colorless semi-solid which was directly used in the
next step. Yield (0.35 g, 70%).
[1335] Step 3: Deprotection of the above mixture following the
method used in Example 12 gave Example 52 hydrochloride as a white
solid. Yield (0.10 g, 36%); .sup.1H NMR (400 MHz, DMSO-d.sub.6+ 5%
D.sub.2O) .delta. 7.34-7.29 (m, 2H), 7.24 (s, 1H), 7.01 (d, J=7.2
Hz, 1H), 3.32 (t, J=6.4 Hz, 2H), 3.12 (t, J=6.4 Hz, 2H), 3.03 (t,
J=7.2 Hz, 2H), 1.66-1.61 (m, 4H), 1.56-1.51 (m, 2H), 1.25-1.04 (m,
7H), 0.87-0.82 (m, 2H); RP-HPLC (Method 4) t.sub.R=6.07 min, 99.37%
(AUC); ESI MS m/z 289.31 [M+H].sup.+.
Example 53
Preparation of
4-((3-(3-amino-1-hydroxypropyl)phenylamino)methyl)heptan-4-ol
##STR00353##
[1337]
4-((3-(3-Amino-1-hydroxypropyl)phenylamino)methyl)heptan-4-ol was
prepared following the method used in Example 67.
[1338] Step 1: Reaction between 2,2-dipropyloxirane and aniline 12
gave
3-hydroxy-3-(3-(2-hydroxy-2-propylpentylamino)phenyl)propanenitrile
as a pale yellow semi-solid. Yield (1.0 g, 40%); .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.17 (t, J=8.0 Hz, 1H), 6.69-6.67 (m, 2H),
6.63 (d, J=8.0 Hz, 1H), 4.96-4.94 (m, 1H), 4.05 (bs, 1H), 3.08 (s,
2H), 2.76 (d, J=6.4 Hz, 2H), 2.36 (bs, 1H), 1.52 (t, J=8.4 Hz, 4H),
1.41-1.32 (m, 4H), 0.94 (t, J=7.2 Hz, 6H).
[1339] Step 2: BH.sub.3-Me.sub.2S reduction of
3-hydroxy-3-(3-(2-hydroxy-2-propylpentylamino)phenyl)propanenitrile
gave Example 53 as a white solid. Yield (0.6 g, 60%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 6.97 (t, J=7.6 Hz, 1H), 6.59 (s,
1H), 6.48 (d, J=8.0, 2H), 4.97 (t, J=5.2 Hz, 1H), 4.52-4.49 (m,
1H), 4.20 (bs,1H), 2.88 (d, J=5.2 Hz, 2H), 2.66-2.62 (m, 2H),
1.65-1.59 (m, 2H), 1.43-1.39 (m, 4H), 1.32-1.24 (m, 4H), 0.84 (t,
J=7.2 Hz, 6H); RP-HPLC (Method 3) t.sub.R=3.87 min, 96.19% (AUC);
ESI MS m/z 295.38 [M+H].sup.+.
Example 54
Preparation of
3-amino-1-(3-(2-hydroxy-2-propylpentylamino)phenyl)propan-1-one
##STR00354##
[1341]
3-Amino-1-(3-(2-hydroxy-2-propylpentylamino)phenyl)propan-1-one is
prepared following the method used in Example 52.
[1342] Step 1: Protection of Example 53 with Boc.sub.2O gives
tert-butyl
3-hydroxy-3-(3-(2-hydroxy-2-propylpentylamino)phenyl)propylcarbamate.
[1343] Step 2: Oxidation of tert-butyl
3-hydroxy-3-(3-(2-hydroxy-2-propylpentylamino)phenyl)propylcarbamate
following the method used in Example 12 gives tert-butyl
3-oxo-3-(3-(2-hydroxy-2-propylpentylamino)phenyl)propylcarbamate.
[1344] Step 3: Deprotection of tert-butyl
3-oxo-3-(3-(2-hydroxy-2-propylpentylamino)phenyl)propylcarbamate
following the method used in Example 12 gives Example 54
hydrochloride.
Example 55
Preparation of
1-((3-(3-amino-1-hydroxypropyl)phenylamino)methyl)cyclohexanol
##STR00355##
[1346]
1-((3-(3-Amino-1-hydroxypropyl)phenylamino)methyl)cyclohexanol was
prepared following the method described below.
[1347] Step 1: TBDMS-Cl (2.7 g, 18.24 mmol) was added at 0.degree.
C. to a stirred solution of aniline 11 (3 g, 15.62 mmol) and TEA
(1.73 g, 17.18 mmoles) in DMF and the reaction mixture was stirred
at RT for 4 h. The reaction mixture was partitioned between EtOAc
and water. Organic layer was washed with water 2.times., dried over
sodium sulfate and concentrated under reduced pressure to give
3-(tert-butyldimethylsilyloxy)-3-(3-nitrophenyl)propanenitrile as
colorless liquid. Yield (4.0 g, 83%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.31 (s, 1H), 8.18 (d, J=7.6 Hz, 1H), 7.88
(d, J=7.6 Hz, 1H), 7.70 (t, J=7.6 Hz, 1H), 5.33 (t, J=5.6 Hz, 1H),
3.01-2.92 (m, 2H), 0.88 (s, 9H), 0.13 (s, 3H), -0.05 (s, 3H).
[1348] Step 2: Hydrogenation of
3-(tert-butyldimethylsilyloxy)-3-(3-nitrophenyl)propanenitrile
following the method used in Example 11 gave
3-(3-aminophenyl)-3-(tert-butyldimethylsilyloxy)propanenitrile as a
colorless oil. Yield (3.5 g, 96%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.97 (t, J=7.6 Hz, 1H), 6.56 (s, 1H), 6.51
(d, J=7.6 Hz, 1H), 6.46 (d, J=7.6 Hz, 1H), 5.07 (s, 2H), 4.87 (t,
J=6.0 Hz, 1H), 2.82-2.70 (m, 2H), 0.86 (s, 9H), 0.07 (s, 3H), -0.06
(s, 3H).
[1349] Step 3: Epoxide ring opening of 2,2-dipropyloxirane with
3-(3-aminophenyl)-3-(tert-butyldimethylsilyloxy)propanenitrile
following the method used in Example 67 gave
3-(tert-butyldimethylsilyloxy)-3-(3-((l-hydroxycyclohexyl)methylamino)phe-
nyl)propanenitrile as a colorless oil. Yield (1.5 g, 56%); .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 7.01 (t, J=7.6 Hz, 1H), 6.63
(s, 1H), 6.57 (d, J=7.6 Hz, 1H), 6.52 (d, J=7.6 Hz, 1H), 5.22 (t,
J=5.2 Hz, 1H), 4.91 (t, J=6.4 Hz, 1H), 4.21 (s, 1H), 2.84 (d, J=5.6
Hz, 2H), 2.78-2.75 (m, 2H), 1.77-1.39 (m, 10H), 0.87 (s, 9H), 0.05
(s, 3H), -0.05 (s, 3H).
[1350] Step 4: BH.sub.3-Me.sub.2S reduction of
3-(tert-butyldimethylsilyloxy)-3-(3-((1-hydroxycyclohexyl)methylamino)phe-
nyl)propanenitrile following the method used in Example 11 gave
crude
1-((3-(3-amino-1-hydroxypropyl)phenylamino)methyl)cyclohexanol
hydrochloride which was taken directly into the next step.
[1351] Step 5: Boc protection of
1-((3-(3-amino-1-hydroxypropyl)phenylamino)methyl)cyclohexanol
hydrochloride following the method used in Example 11 gave
tert-butyl
3-hydroxy-3-(3-((1-hydroxycyclohexyl)methyl-amino)phenyl)propylcarbamate
as a colorless oil. Yield (0.6 g, 29%, after two steps); .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 6.96 (t, J=7.6 Hz, 1H), 6.57
(s, 1H), 6.48-6.44 (m, 2H), 5.09 (t, J=5.6 Hz, 1H), 5.0 (d, J=4.4
Hz, 1H), 4.38-4.37 (m, 1H), 4.17 (s, 1H), 2.95-2.91 (m, 4H),
1.65-1.61 (m, 2H), 1.57-1.52 (m, 5H), 1.41-1.36 (m, 5H), 1.36 (s,
9H).
[1352] Step 6: tert-Butyl
3-hydroxy-3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)propylcarbamate
was deprotected following the method used in Example 24.
Purification by column chromatography (5% NH.sub.4OH/10%
MeOH/CH.sub.2Cl.sub.2) gave Example 55 as a pale yellow oil. Yield
(0.3 g, 86%); .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.08 (t,
J=7.6 Hz, 1H), 6.69 (s, 1H), 6.61 (d, J=7.6 Hz, 1H), 6.59 (d, J=7.6
Hz, 1H), 4.76-4.66 (m, 1H), 3.07 (s, 2H), 3.05-2.97 (m, 2H),
2.03-1.96 (m, 2H), 1.68-1.61 (m, 5H), 1.59-1.49 (m, 5H); RP-HPLC
(Method 6) t.sub.R=4.06 min, 88.6% (AUC); ESI MS m/z 279.30
[M+H].sup.+.
Example 56
Preparation of
3-amino-1-(3-((1-hydroxycyclohexyl)methylamino)phenyl)propan-1-one
##STR00356##
[1354]
3-Amino-1-(3-((1-hydroxycyclohexyl)methylamino)phenyl)propan-1-one
is prepared following the method used in Example 54.
[1355] Step 1: Protection of Example 55 with Boc.sub.2O gives
tert-butyl
3-hydroxy-3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)propylcarbamate.
[1356] Step 2: Oxidation of tert-butyl
3-hydroxy-3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)propylcarbamate
following the method used in Example 12 gives tert-butyl
3-oxo-3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)propylcarbamate.
[1357] Step 3: Deprotection of tert-butyl
3-oxo-3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)propylcarbamate
following the method used in Example 12 gives Example 56
hydrochloride.
Example 57
Preparation of
N-(3-(3-amino-2,2-dideutero-1-hydroxypropyl)phenyl)cyclohexanecarboxamide
##STR00357##
[1359]
N-(3-(3-amino-2,2-dideutero-1-hydroxypropyl)phenyl)cyclohexanecarbo-
xamide was prepared following the methods used in Examples 82, 5,
115, 15, 12.
[1360] Step 1: Hydrogenation of
2,2-dideutero-3-hydroxy-3-(3-nitrophenyl)propanenitrile was done
following the method used in Example 5 for 48 hrs to give crude
3-(3-aminophenyl)-2,2-dideutero-3-hydroxypropanenitrile as a
colorless oil which was directly used in the next step without
further purification.
[1361] Step 2: BH.sub.3-Me.sub.2S reduction of crude
3-(3-aminophenyl)-2,2-dideutero-3-hydroxypropanenitrile following
the method used in Example 115 gave
3-amino-1-(3-aminophenyl)-2,2-dideuteropropan-1-ol hydrochloride as
a colorless oil which was directly used in the next step without
further purification.
[1362] Step 3: Boc protection of
3-amino-1-(3-aminophenyl)-2,2-dideuteropropan-1-ol hydrochloride
following the method used in Example 15 gave, after purification by
column chromatography (66% to 75% EtOAc-hexanes gradient)
tert-butyl
(3-(3-aminophenyl)-2,2-dideutero-3-hydroxypropyl)carbamate as a
colorless oil. Yield (0.500 g, 18% after 3 steps); .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 6.90 (t, J=7.6 Hz, 1H), 6.69 (t, J=5.6
Hz, 1H), 6.50 (t, J=1.6 Hz, 1H), 6.36-6.41 (m, 2H), 4.96 (d, J=4.0
Hz, 1H), 4.94 (s, 2H), 4.33 (d, J=4.0 Hz, 1H), 2.91 (t, J=4.8 Hz,
2H), 1.32 (s, 9H).
[1363] Step 4: Reaction between tert-butyl
(3-(3-aminophenyl)-2,2-dideutero-3-hydroxypropyl)carbamate and
chloride 36 following the method used in Example 15 gave tert-butyl
(3-(3-(cyclohexanecarboxamido)phenyl)-2,2-dideutero-3-hydroxypropyl)carba-
mate. Yield (0.230 g, 65%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.73 (s, 1H), 7.54 (s, 1H), 7.43 (d, J=8.8 Hz, 1H), 7.16
(t, J=7.6 Hz, 1H), 6.91 (t, J=5.6 Hz, 1H), 6.73 (t, J=5.6 Hz, 1H),
5.16 (d, J=4.0 Hz, 1H), 4.44 (d, J=4.4 Hz, 1H), 2.92 (d, J=5.6 Hz,
2H), 2.24-2.32 (m, 1H), 1.58-1.80 (m, 6H), 1.30-1.41 (m, 11H),
1.12-1.28 (m, 3H).
[1364] Step 5: Deprotection of tert-butyl
(3-(3-(cyclohexanecarboxamido)phenyl)-2,2-dideutero-3-hydroxypropyl)carba-
mate following the method used in Example 12 gave Example 57
hydrochloride as a white solid. Yield (0.170 g, 89%); .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 7.01 (d, J=2.4 Hz, 1H), 7.26-7.33 (m,
2H), 7.09-7.11 (m, 1H), 4.78 (s, 1H), 2.99-3.11 (m, 2H), 2.31-2.40
(m, 1H), 1.81-1.84 (m, 4H), 1.68-1.76 (m, 1H), 1.46-1.57 (m, 2H),
1.22-1.42 (m, 3H).
Example 58
Preparation of
N-(3-(3-amino-2,2-dideutero-1-hydroxypropyl)phenyl)cyclohexanesulfonamide
##STR00358##
[1366]
N-(3-(3-amino-2,2-dideutero-1-hydroxypropyl)phenyl)cyclohexanesulfo-
namide was prepared following the method used in Examples 57,
5.
[1367] Step 1: Sulfonation of tert-butyl
(3-(3-aminophenyl)-2,2-dideutero-3-hydroxypropyl)carbamate by
sulfonyl chloride 8 following the method used in Example 5 gave,
after purification by column chromatography (EtOAc-hexanes, 2:1)
tert-butyl
(3-(3-(cyclohexanesulfonamido)phenyl)-2,2-dideutero-3-hydroxypropyl)carba-
mate as a colorless oil. Yield (0.170 g, 44%); .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.66 (s, 1H), 7.16-7.22 (m, 2H),
6.99-7.06 (m, 1H), 6.97 (d, J=8.0 Hz, 1H), 6.73 (t, J=5.6 Hz, 1H),
5.20 (d, J=4.4 Hz, 1H), 4.47 (d, J=4.4 Hz, 1H), 2.86-2.95 (m, 3H),
1.96-1.98 (m, 2H), 1.68-1.76 (m, 2H), 1.51-1.58 (m, 1H), 1.25-1.42
(m, 11H), 1.02-1.18 (m, 3H).
[1368] Step 2: Step 3: Deprotection of tert-butyl
(3-(3-(cyclohexanesulfonamido)phenyl)-2,2-dideutero-3-hydroxypropyl)carba-
mate following the method used in Example 57 gave Example 58
hydrochloride as a white solid. Yield (0.188 g, quant.); .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 7.35 (t, J=2.0 Hz, 1H), 7.29 (t,
J=8.0 Hz, 1H), 7.09-7.12 (m, 2H), 2.92-3.11 (m, 3H), 2.07-2.22 (m,
2H), 1.80-1.88 (m, 2H), 1.62-1.68 (m, 1H), 1.46-1.58 (m, 2H),
1.14-1.28 (m, 3H).
Example 59
Preparation of
3-amino-1-(3-(3-phenylpropylamino)phenyl)propan-1-ol
##STR00359##
[1370] 3-Amino-1-(3-(3-phenylpropylamino)phenyl)propan-1-ol was
prepared following the method used in Example 11.
[1371] Step 1: Hydrogenation of nitrobenzene 11 and
3-phenylpropanal gave
3-hydroxy-3-(3-(3-phenylpropylamino)phenyl)propanenitrile as a
yellow oil. Yield (1.0 g, 68%); .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.32-7.28 (m, 2H), 7.22-7.18 (m, 3H), 7.16-7.14 (m, 1H),
6.66 (d, J=7.6 Hz, 1H), 6.57 (s, 1H), 6.53 (d, J=8.0 Hz, 1H), 4.93
(m, 1H), 3.73 (bs, 1H), 3.15 (t, J=7.2 Hz, 2H), 2.75-2.72 (m, 4H),
2.22 (d, J=3.2 Hz, 1H), 1.99-1.92 (quintet, J=7.2 Hz, 2H).
[1372] Step 2: BH.sub.3-Me.sub.2S reduction of
3-hydroxy-3-(3-(3-phenylpropylamino)phenyl)propanenitrile gave
Example 59 hydrochloride as a yellow solid. Yield (0.85 g, 84%);
.sup.1H NMR (400 MHz, DMSO-d.sub.6+5% D.sub.2O) .delta. 7.38 (t,
J=8.0 Hz, 1H), 7.29-7.23 (m, 3H), 7.18 (d, J=7.2 Hz, 4H), 7.13 (d,
J=7.6 Hz, 1H), 4.69-4.66 (m, 1H), 3.19 (t, J=8.0 Hz, 2H), 2.85 (m,
2H), 2.65 (t, J=7.6 Hz, 2H), 1.94-1.81 (m, 4H); RP-HPLC (Method 5)
t.sub.R=5.04 min, 94.44% (AUC); ESI MS m/z 285.38 [M+H].sup.+.
Example 60
Preparation of
3-amino-1-(3-(3-phenylpropylamino)phenyl)propan-1-one
##STR00360##
[1374] 3-Amino-1-(3-(3-phenylpropylamino)phenyl)propan-1-one was
prepared following the method used in Example 52.
[1375] Step 1: Protection of Example 59 with Boc.sub.2O gave a
mixture of tert-butyl
(3-hydroxy-3-(3-((3-phenylpropyl)amino)phenyl)propyl)carbamate and
tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)-1-hydroxypropyl)phenyl)(3-phenylpropyl-
)carbamate which was directly used in the next step without
purification.
[1376] Step 2: Oxidation of the above mixture gave a mixture of
tert-butyl
(3-oxo-3-(3-((3-phenylpropyl)amino)phenyl)propyl)carbamate and
tert-butyl
(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)(3-phenylpropyl)carbam-
ate which was directly used in the next step without
purification.
[1377] Step 3: Deprotection of the above mixture gave Example 60
hydrochloride as an off-white solid. Yield (0.30 g, 51%); .sup.1H
NMR (400 MHz, DMSO-d.sub.6+5% D.sub.2O) .delta. 7.61-7.59 (m, 2H),
7.48 (t, J=8.0 Hz, 1H), 7.33 (d, J=8.0 Hz, 1H), 7.28-7.25 (m, 2H),
7.20-7.15 (m, 3H), 3.37 (t, J=6.0 Hz, 2H), 3.19-3.11 (m, 4H), 2.66
(t, J=7.6 Hz, 2H), 1.93-1.87 (quintet, J=7.2 Hz, 2H). RP-HPLC
(Method 5) t.sub.R=4.45 min, 96.38% (AUC); ESI MS m/z 283.25
[M+H].sup.+.
Example 61
Preparation of
3-amino-1-(3-((4,4-difluorocyclohexyl)methylamino)phenyl)propan-1-ol
##STR00361##
[1379]
3-Amino-1-(3-(4,4-difluorocyclohexyl)methylamino)phenyl)propan-1-ol
is prepared following the method used in Example 37.
[1380] Step 1: Hydrogenation of nitrobenzene 11 and
4,4-difluorocyclohexanecarbaldehyde gives
3-(3-(4,4-difluorocyclohexyl)methylamino)phenyl)-3-hydroxypropanenitrile.
[1381] Step 2: BH.sub.3-Me.sub.2S reduction of
3-(3-((4,4-difluorocyclohexyl)methylamino)phenyl)-3-hydroxypropanenitrile
gives Example 61.
Example 62
Preparation of
3-amino-1-(3-((4,4-difluorocyclohexyl)methylamino)phenyl)propan-1-one
##STR00362##
[1383]
3-Amino-1-(3-(4,4-difluorocyclohexyl)methylamino)phenyl)propan-1-on-
e is prepared following the method used in Example 38.
[1384] Step 1: Protection of Example 61 with Boc.sub.2O gives
tert-butyl
3-(3-((4,4-difluorocyclohexyl)methylamino)phenyl)-3-hydroxypropylcarbamat-
e.
[1385] Step 2: Oxidation tert-butyl
3-(3-((4,4-difluorocyclohexyl)methylamino)phenyl)-3-hydroxypropylcarbamat-
e by MnO.sub.2 gives tert-butyl
3-(3-((4,4-difluorocyclohexyl)methylamino)phenyl)-3-oxopropylcarbamate.
[1386] Step 3: Deprotection of tert-butyl
3-(3-((4,4-difluorocyclohexyl)methylamino)phenyl)-3-oxopropylcarbamate
gives Example 62 hydrochloride.
Example 63
Preparation of
3-(3-aminopropyl)-N-((4,4-difluorocyclohexyl)methyl)aniline
##STR00363##
[1388] 3-(3-Aminopropyl)-N-((4,4-difluorocyclohexyl)methyl)aniline
is prepared following the method used in Example 31.
[1389] Step 1: Hydrogenation of nitrobenzene 40 and
4,4-difluorocyclohexanecarbaldehyde gives
2-(3-(3-((4,4-difluorocyclohexyl)methylamino)phenyl)propyl)isoindoline-1,-
3-dione.
[1390] Step 2: Deprotection of
2-(3-(3-((4,4-difluorocyclohexyl)methylamino)phenyl)propyl)isoindoline-1,-
3-dione gives Example 63.
Example 64
Preparation of 3-(3-aminopropyl)-N-(3-phenylpropyl)aniline E
##STR00364##
[1392] 3-(3-Aminopropyl)-N-(3-phenylpropyl)aniline was prepared
following the method used in Example 33.
[1393] Step 1: A mixture of
2,2,2-trifluoro-N-(3-(3-nitrophenyl)allyl)acetamide (1.0 g, 3.6
mmol) and 3-phenylpropanal (0.48 g, 3.6 mmol) in EtOAc was degassed
and saturated with argon. 10% Pd/C (500 mg) was added to this
solution and the resulting mixture was stirred under H.sub.2 at 1
atm for 16 hrs, filtered through Celite, and concentrated under
reduced pressure. Purification by flash chromatography (40% to 50%
EtOAc-hexanes gradient) gave
2,2,2-trifluoro-N-(3-(3-(3-phenylpropylamino)phenyl)propyl)acetamide
as a colorless semi-solid. Yield (0.54 g, 41%)..sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.41 (br.s, 1H), 7.30-7.17 (m, 5H) 6.95
(t, J=7.6 Hz, 1H), 6.36-6.34 (m, 3H), 5.50 (t, J=5.6 Hz, 1H), 3.18
(q, J=6.4 Hz, 2H), 2.98 (q, J=6.4 Hz, 2H), 2.67 (t, J=8.0 Hz, 2H),
2.43 (t, J=7.6 Hz, 2H), 1.82 (quintet, J=7.6 Hz, 2H), 1.73
(quintet, J=7.6 Hz, 2H).
[1394] Step 2: A mixture of
2,2,2-trifluoro-N-(3-(3-(3-phenylpropylamino)phenyl)propyl)acetamide
(0.54 g, 1.4 mmol) and K.sub.2CO.sub.3 (0.73 g, 5.3 mmol) in
MeOH:H.sub.2O was stirred at room temperature for 24 hours. The
solvent was removed under reduced pressure. Purification by flash
chromatography (5% to 6% MeOH-CH.sub.2Cl.sub.2 gradient) gave
Example 64 as a light green solid. Yield (0.22 g, 55%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.28-7.20 (m, 4H), 7.17 (t, J=7.2
Hz, 1H), 6.94 (t, J=7.6 Hz, 1H), 6.35-6.32 (m, 3H), 5.54 (t, J=5.6
Hz, 1H), 2.96 (q, J=6.4 Hz, 2H), 2.70-2.63 (m, 4H), 2.47-2.43 (m,
2H), 1.80 (quintet, J=7.6 Hz, 2H), 1.73 (quintet, J=7.6 Hz, 2H);
RP-HPLC (Method-3) t.sub.R=3.95 min, 94.30% (AUC); ESI MS m/z
269.25 [M+H].sup.+.
Example 65
Preparation of 3-(3-aminopropyl)-N-(5-methoxypentyl)aniline
##STR00365##
[1396] 3-(3-Aminopropyl)-N-(5-methoxypentyl)aniline is prepared
following the method used in Example 31.
[1397] Step 1: Hydrogenation of nitrobenzene 40 and
5-methoxypentanal gives
2-(3-(3-(5-methoxypentylamino)phenyl)propyl)isoindoline-1,3-dione.
[1398] Step 2: Deprotection of
2-(3-(3-(5-methoxypentylamino)phenyl)propyl)isoindoline-1,3-dione
gives Example 65.
Example 66
Preparation of 5-(3-(3-aminopropyl)phenylamino)pentan-1-ol
##STR00366##
[1400] 5-(3-(3-Aminopropyl)phenylamino)pentan-1-ol is prepared
following the method used in Example 31.
[1401] Step 1: Hydrogenation of nitrobenzene 40 and
5-hydroxypentanal gives
2-(3-(3-(5-hydroxypentylamino)phenyl)propyl)isoindoline-1,3-dione.
[1402] Step 2: Deprotection of
2-(3-(3-(5-hydroxypentylamino)phenyl)propyl)isoindoline-1,3-dione
gives Example 66.
Example 67
Preparation of
4-((3-(3-aminopropyl)phenylamino)methyl)heptan-4-ol
##STR00367##
[1404] 4-((3-(3-Aminopropyl)phenylamino)methyl)heptan-4-ol was
prepared following the method described below.
[1405] Step 1: To a stirred solution of
2-(3-(3-aminophenyl)propyl)isoindoline-1,3-dione (0.50 g, 1.78
mmol) in EtOH:H.sub.2O (9:1), 2,2-dipropyloxirane (0.45 g, 3.57
mmol) was added and the reaction mixture was stirred under reflux
for 36 h. The reaction mixture was concentrated under reduced
pressure. Purification by column chromatography (20% to 30%
EtOAc-hexanes gradient) gave
2-(3-(3-(2-hydroxy-2-propylpentylamino)phenyl)propyl)isoindoline-1,3-dion-
e as a yellow semisolid. Yield (0.22 g, 30%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.86-7.81 (m, 4H), 6.92 (t, J=7.8 Hz, 1H),
6.45 (s, 1H), 6.41 (d, J=8.0 Hz, 1H), 6.37 (d, J=7.6 Hz, 1H), 4.89
(bs, 1H), 4.16 (s, 1H), 3.59 (t, J=7.2 Hz, 2H), 2.87 (d, J=5.2,
2H), 2.46-2.50 (m, 2H), 1.83-1.90 (quintet, J=7.2 Hz, 2H),
1.42-1.38 (m, 4H), 1.28-1.24 (m, 4H), 0.84 (t, J=7.2 Hz, 6H).
[1406] Step 2: A mixture of
2-(3-(3-(2-hydroxy-2-propylpentylamino)phenyl)propyl)isoindoline-1,3-dion-
e (0.22 g, 0.71 mmol) and hydrazine hydrate (0.1 ml, 1.6 mmol) in
ethanol was stirred at room temperature for 24 hours. The solvent
was evaporated under reduced pressure. Purification by column
chromatography (5% to 10% MeOH-CH2Cl2 gradient) gave
4-((3-(3-aminopropyl)phenylamino)methyl)heptan-4-ol as a light
yellow semisolid. Yield (0.06 g, 18%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.93 (t, J=7.6 Hz, 1H), 6.44-6.41 (m, 2H),
6.35 (d, J=7.6 Hz, 1H), 4.92 (t, J=5.2 Hz, 1H), 4.19 (bs, 1H), 2.87
(d, J=5.2 Hz, 2H), 2.54-2.50 (m, 2H), 2.44 (t, J=7.6 Hz, 2H),
1.55-1.62 (quintet, J=7.2 Hz, 2H), 1.42-1.33 (m, 4H), 1.32-1.27 (m,
4H), 0.85 (t, J=7.2 Hz, 6H); RP-HPLC (Method 3) t.sub.R=4.44 min,
97.48% (AUC); ESI MS m/z 279.31 [M+H].sup.+.
Example 68
Preparation of
3-((3-(3-aminopropyl)phenylaminomethyl)pentan-3-ol
##STR00368##
[1408] 3-((3-(3-Aminopropyl)phenylamino)methyl)pentan-3-ol is
prepared following the method used in Example 67.
[1409] Step 1: Reaction between 2,2-diethyloxirane and
2-(3-(3-aminophenyl)propyl)isoindoline-1,3-dione gives
2-(3-(3-((2-ethyl-2-hydroxybutyl)amino)phenyl)propyl)isoindoline-1,3-dion-
e.
[1410] Step 2: Deprotection of
2-(3-(3-(2-ethyl-2-hydroxybutyl)amino)phenyl)propyl)isoindoline-1,3-dione
gives Example 68.
Example 69
Preparation of
1-((3-(3-aminopropyl)phenylamino)methyl)cyclohexanol
##STR00369##
[1412] 1-((3-(3-Aminopropyl)phenylamino)methyl)cyclohexanol was
prepared following the method used in Example 29.
[1413] Step 1: Epoxide ring opening of 1-oxaspiro[2.5]octane with
N-(3-(3-aminophenyl)propyl)-2,2,2-trifluoroacetamide gave
2,2,2-trifluoro-N-(3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)propyl)a-
cetamide as a colorless oil. Yield (0.8 g, 46%); .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.42 (bs, 1H), 6.94 (t, J=8.0 Hz, 1H),
6.44-6.37 (m, 2H), 6.35-6.33 (d, J=7.6 Hz, 1H), 5.07 (t, J=5.6 Hz,
1H), 4.18 (s, 1H), 3.18 (q, J=6.4 Hz, 2H), 2.91 (d, J=5.6 Hz, 2H),
2.44-2.42 (m, 2H), 1.76-1.70 (m, 2H), 1.58-1.49 (m, 6H), 1.41-1.27
(m, 4H).
[1414] Step 2: A mixture of
2,2,2-trifluoro-N-(3-(3-((1-hydroxycyclohexyl)methylamino)phenyl)propyl)a-
cetamide 2 (0.7 g, 1.9 mmol) and K.sub.2CO.sub.3 (0.815 g, 5.8
mmol) in MeOH:H.sub.2O (1:1) was stirred at room temperature for 16
h. The solvent was evaporated under reduced pressure. The residue
was partitioned between DCM and water. Aqueous layer was extracted
five times with DCM. Combined organic layers were dried over
anhydrous sodium sulfate and concentrated under reduced pressure.
Purification by flash chromatography (5% to 6% MeOH-DCM+5%
NH.sub.4OH) gave crude which was dissolved in dioxane and stirred
with 4M HCl in Dioxane. The mixture was concentrated under reduced
pressure and triturated with diethyl ether to give Example 69
hydrochloride as a white solid. Yield (0.32 g, 56%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.76 (m, 3H), 6.96 (t, J=7.6 Hz,
1H), 6.45 (m, 2H), 6.34 (d, J=7.6 Hz, 1H), 5.13 (t, J=5.6 Hz, 1H),
4.20 (s, 1H), 2.91 (d, J=5.6 Hz, 2H), 2.76 (t, J=7.6 Hz, 2H),
2.50-2.46 (m, 2H), 1.83-1.75 (m, 2H), 1.69-1.56 (m, 6H), 1.53-1.38
(m, 4H); RP-HPLC (Method 6) t.sub.R=4.57 min, 92.1% (AUC); ESI MS
m/z 263.2 [M+H].sup.+.
Example 70
Preparation of
1-((3-(3-aminopropyl)phenylamino)methyl)cyclopentanol
##STR00370##
[1416] 1-((3-(3-Aminopropyl)phenylamino)methyl)cyclopentanol is
prepared following the method used in Example 67.
[1417] Step 1: Reaction between 1-oxaspiro[2.4]heptane and
2-(3-(3-aminophenyl)propyl)isoindoline-1,3-dione gives
2-(3-(3-(((1-hydroxycyclopentyl)methyl)amino)phenyl)propyl)isoindoline-1,-
3-dione.
[1418] Step 2: Deprotection of
2-(3-(3-((1-hydroxycyclopentyl)methyl)amino)phenyl)propyl)isoindoline-1,3-
-dione gives Example 68.
Example 71
Preparation of N-(3-(3-aminopropyl)phenyl)-2-propylpentanamide
##STR00371##
[1420] N-(3-(3-Aminopropyl)phenyl)-2-propylpentanamide was prepared
following the method described below.
[1421] Step 1: Et.sub.3N (3.24 mL, 23.25 mmol) was added to a
solution of 2-propylpentanoic acid in (2 g, 11.62 mmol) in DMF. The
reaction mixture was cooled to 0.degree. C. HATU (6.63 g, 17.4
mmol) was added to the reaction mixture which was stirred for 15
min and then 3-bromoaniline (2.5 g, 17.43 mmol) was added. The
reaction mixture was stirred for 2 h at 0.degree. C. The reaction
mixture was diluted with H.sub.2O, extracted with EtOAc, and
organic layer was concentrated under reduced pressure. The residue
was washed with pentane to give
N-(3-bromophenyl)-2-propylpentanamide as a white solid. Yield (1.6
g, 47%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.82 (s, 1H),
7.44 (d, J=8.0 Hz, 1H), 7.23 (d, J=7.6 Hz, 1H), 7.17 (t, J=8.0 Hz,
1H), 7.12 (bs, 1H), 2.21-2.14 (m, 1H), 1.73-1.63 (m, 2H), 1.51-1.45
(m, 2H), 1.43-1.25 (m, 4H), 0.92 (t, J=7.2 Hz, 6H).
[1422] Step 2: Et.sub.3N (1.2 mL) was added to a solution of
N-(3-bromophenyl)-2-propylpentanamide (0.6 g, 2.01 mmol),
tert-butyl allylcarbamate (1.026 g, 6.55 mmol) and P(o-tol).sub.3
(0.06 g, 0.201 mmol) in DMF (10 mL). The reaction mixture was
degassed for 30 min and then added Pd(OAc).sub.2 (0.09 g, 0.409
mmol) was added. The reaction mixture was again degassed for 15 min
and then refluxed at 90.degree. C. for 8 h. The reaction mixture
was diluted with EtOAc, washed with H.sub.2O, brine. The organic
layer was concentrated under reduced pressure. Purification by
column chromatography (100-200 mesh silica, elution 10% to 15%
EtOAc in hexane) gave (E)-tert-butyl
3-(3-(2-propylpentanamido)phenyl)allylcarbamate as a yellow oil.
Yield (0.7 g, 37%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.65
(s, 1H), 7.43 (t, J=7.6 Hz, 1H), 7.24 (s, 1H), 7.08 (dd, J=1.2, 7.2
Hz, 2H), 6.48 (d, J=16.0 Hz, 1H), 6.26-6.16 (m, 1H), 4.67 (bs, 1H),
3.89 (bs, 2H), 2.17-2.04 (m, 1H), 1.73-1.64 (m, 4H), 1.48 (s, 9H),
1.45-1.14 (m, 4H), 0.91 (t, J=7.2 Hz, 6H).
[1423] Step 3: A solution of (E)-tert-butyl
3-(3-(2-propylpentanamido)phenyl)allylcarbamate (0.5 g, 1.32 mmol)
in ethanol was degassed by bubbling argon for 2 min. Pd/C (10% wt,
0.5 g) was added and the reaction mixture atmosphere was changed to
hydrogen by alternating between vacuum and hydrogen 2.times.. The
reaction mixture was stirred under a H.sub.2-filled balloon for 16
h then filtered through Celite and the filtrate was concentrated
under reduced pressure. Purification by column chromatography
(100-200 mesh silica, 10% to 15% EtOAc in hexane) gave compound
tert-butyl 3-(3-(2-propylpentanamido)phenyl)propylcarbamate as a
thick yellow oil. Yield (0.5 g, 99%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.45 (s, 1H), 7.31 (d, J=7.6 Hz, 1H), 7.22 (t,
J=8.0 Hz, 1H), 7.08 (bs, 1H), 6.93 (d, J=7.6 Hz, 1H), 4.60 (bs,
1H), 3.15-3.14 (m, 2H), 2.62 (t, J=7.2 Hz, 2H), 2.49-2.16 (m, 1H),
1.82-1.72 (m, 2H), 1.72-1.66 (m, 2H), 1.37 (s, 9H), 1.36-1.22 (m,
6H), 0.92 (t, J=7.2 Hz, 6H).
[1424] Step 4: 4M HCl/dioxane was added to a solution of tert-butyl
3-(3-(2-propylpentanamido)phenyl)propylcarbamate in DCM. The
reaction mixture was stirred for 30 min. The reaction mixture was
concentrated under reduced pressure to give Example 71 as a white
solid. Yield (0.142 g, 41%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.89 (s, 1H), 7.93 (bs, 3H), 7.58 (s, 1H), 7.39 (d, J=8.0
Hz, 1H), 7.20 (t, J=7.6 Hz, 1H), 6.88 (d, J=7.6 Hz, 1H), 2.79-2.77
(m, 2H), 2.60 (t, J=7.6 Hz, 2H), 2.42-2.38 (m, 1H), 1.83 (quintet,
J=7.6 Hz, 2H), 1.56-1.48 (m, 2H), 1.37-1.19 (m, 6H), 0.86 (t, J=7.2
Hz, 6H); RP-HPLC (Method 6) t.sub.R=5.01 min, 99.58% (AUC); ESI MS
m/z 277.30 [M+H].sup.+.
Example 72
Preparation of N-(3-(3-aminopropyl)phenyl)heptane-4-sulfonamide
##STR00372##
[1426] N-(3-(3-Aminopropyl)phenyl)heptane-4-sulfonamide is prepared
following the method used in Example 6.
[1427] Step 1: Sulfonation of aniline 17 by heptane-4-sulfonyl
chloride following the method used in Example 6 gives tert-butyl
3-(3-(1-propylbutylsulfonamido)phenyl)propylcarbamate.
[1428] Step 2: Deprotection of tert-butyl
3-(3-(1-propylbutylsulfonamido)phenyl)propylcarbamate gave Example
72 hydrochloride.
Example 73
Preparation of
N-(3-(3-amino-1-hydroxypropyl)phenyl)-2-propylpentanamide
##STR00373##
[1430] N-(3-(3-Amino-1-hydroxypropyl)phenyl)-2-propylpentanamide is
prepared following the method used in Example 15.
[1431] Step 1: Acylation of aniline 35 by 2-propylpentanoyl
chloride following the method used in Example 15 gives tert-butyl
3-hydroxy-3-(3-(2-propylpentanamido)phenyl)propylcarbamate.
[1432] Step 2: Deprotection of tert-butyl
3-hydroxy-3-(3-(2-propylpentanamido)phenyl)propylcarbamate gives
Example 73 hydrochloride.
Example 74
Preparation of
N-(3-(3-amino-1-hydroxypropyl)phenyl)heptane-4-sulfonamide
##STR00374##
[1434] N-(3-(3-Amino-1-hydroxypropyl)phenyl)heptane-4-sulfonamide
is prepared following the method used in Example 5.
[1435] Step 1: Sulfonation of aniline 35 by heptane-4-sulfonyl
chloride following the method used in Example 5 gives
N-(3-(2-cyano-1-hydroxyethyl)phenyl)heptane-4-sulfonamide.
[1436] Step 2: BH.sub.3-Me.sub.2S reduction of
N-(3-(2-cyano-1-hydroxyethyl)phenyl)heptane-4-sulfonamide following
the method used in Example 5 gives Example 74.
Example 75
Preparation of
N-(3-(3-aminopropanoyl)phenyl)-2-propylpentanamide
##STR00375##
[1438] N-(3-(3-Aminopropanoyl)phenyl)-2-propylpentanamide is
prepared following the method used in Examples 73 16 and 12.
[1439] Step 1: Oxidation of tert-butyl
3-hydroxy-3-(3-(2-propylpentanamido)phenyl)propylcarbamate by PCC
following the method used in Example 16 gives tert-butyl
3-oxo-3-(3-(2-propylpentanamido)phenyl)propylcarbamate.
[1440] Step 2: tert-Butyl
3-oxo-3-(3-(2-propylpentanamido)phenyl)propylcarbamate is
deprotected following the method used in Example 12 to give Example
75 hydrochloride.
Example 76
Preparation of
N-(3-(3-aminopropanoyl)phenyl)heptane-4-sulfonamide
##STR00376##
[1442] N-(3-(3-Aminopropanoyl)phenyl)heptane-4-sulfonamide is
prepared following the methods used in Examples 20, 16, 12.
[1443] Step 1: Protection of Example 74 with Boc.sub.2O following
the method used in Example 20 gives tert-butyl
3-hydroxy-3-(3-(1-propylbutylsulfonamido)phenyl)propylcarbamate.
[1444] Step 2: Oxidation of tert-butyl
3-hydroxy-3-(3-(1-propylbutylsulfonamido)phenyl)propylcarbamate by
PCC following the method used in Example 16 gives tert-butyl
3-oxo-3-(3-(1-propylbutylsulfonamido)phenyl)propylcarbamate.
[1445] Step 3: Deprotection of tert-butyl
3-oxo-3-(3-(1-propylbutylsulfonamido)phenyl)propylcarbamate
following the method used in Example 12 gives Example 76
hydrochloride.
Example 77
Preparation of
3-((3-(3-amino-1-hydroxypropyl)phenylamino)methyl)pentan-3-ol
##STR00377##
[1447]
3-((3-(3-Amino-1-hydroxypropyl)phenylamino)methyl)pentan-3-ol is
prepared following the method used in Example 53.
[1448] Step 1: Reaction between 2,2-diethyloxirane and aniline 12
gives
3-(3-(2-ethyl-2-hydroxybutylamino)phenyl)-3-hydroxypropanenitrile.
[1449] Step 2: BH.sub.3-Me.sub.2S reduction of
3-(3-(2-ethyl-2-hydroxybutylamino)phenyl)-3-hydroxypropanenitrile
gives Example 77.
Example 78
Preparation of
1-((3-(3-amino-1-hydroxypropyl)phenylamino)methyl)cyclopentanol
##STR00378##
[1451]
1-((3-(3-Amino-1-hydroxypropyl)phenylamino)methyl)cyclopentanol is
prepared following the method used in Example 53.
[1452] Step 1: Reaction between 1-oxaspiro[2.4]heptane and aniline
12 gives
3-hydroxy-3-(3-((l-hydroxycyclopentyl)methylamino)phenyl)propanenit-
rile.
[1453] Step 2: BH.sub.3-Me.sub.2S reduction of
3-hydroxy-3-(3-((1-hydroxycyclopentyl)methylamino)phenyl)propanenitrile
gives Example 78.
Example 79
Preparation of
3-amino-1-(3-(2-ethyl-2-hydroxybutylamino)phenyl)propan-1-one
##STR00379##
[1455]
3-Amino-1-(3-(2-ethyl-2-hydroxybutylamino)phenyl)propan-1-one is
prepared following the method used in Example 40.
[1456] Step 1: Protection of Example 77 with Boc.sub.2O gives
tert-butyl
3-(3-(2-ethyl-2-hydroxybutylamino)phenyl)-3-hydroxypropylcarbamate.
[1457] Step 2: Oxidation of tert-butyl
3-(3-(2-ethyl-2-hydroxybutylamino)phenyl)-3-hydroxypropylcarbamate
gives tert-butyl
3-(3-(2-ethyl-2-hydroxybutylamino)phenyl)-3-oxopropylcarbamate.
[1458] Step 3: Deprotection of tert-butyl
3-(3-(2-ethyl-2-hydroxybutylamino)phenyl)-3-oxopropylcarbamate
gives Example 79 hydrochloride.
Example 80
Preparation of
3-amino-1-(3-((1-hydroxycyclopentyl)methylamino)phenyl)propan-1-one
##STR00380##
[1460]
3-Amino-1-(3-((l-hydroxycyclopentyl)methylamino)phenyl)propan-1-one
is prepared following the method used in Example 40.
[1461] Step 1: Protection of Example 78 with Boc.sub.2O gives
tert-butyl
3-hydroxy-3-(3-((1-hydroxycyclopentyl)methylamino)phenyl)propylcarbamate.
[1462] Step 2: Oxidation of tert-butyl
3-hydroxy-3-(3-((1-hydroxycyclopentyl)methylamino)phenyl)propylcarbamate
gives tert-butyl
3-oxo-3-(3-((1-hydroxycyclopentyl)methylamino)phenyl)propylcarbamate.
[1463] Step 3: Deprotection of tert-butyl
3-oxo-3-(3-((1-hydroxycyclopentyl)methylamino)phenyl)propylcarbamate
gives Example 80 hydrochloride.
Example 81
Preparation of
3-amino-1-(3-(cyclohexylmethylamino)phenyl)-1-deuteropropan-1-ol
##STR00381##
[1465]
3-Amino-1-(3-(cyclohexylmethylamino)phenyl)-1-deuteropropan-1-ol
was prepared following the method used in Example 20.
[1466] Step 1: NaBD.sub.4 (0.08 g, 0.94 mmol) was added at
0.degree. C. to a solution of ketone 33 (0.19 g. 0.47 mmol) in
i-PrOH. The reaction mixture was stirred at 0.degree. C. for 2 hr
and then at room temperature for 3 hrs. The reaction mixture was
partitioned between aqueous NH.sub.4Cl and ethyl acetate, dried
over anhydrous Na.sub.2SO.sub.4 and concentrated under reduced
pressure to give tert-butyl
3-(3-(cyclohexylmethylamino)phenyl)-3-deutero-3-hydroxypropylcarbamate
as a colorless oil which was used directly in the next step.
[1467] Step 2: Deprotection of tert-butyl
3-(3-(cyclohexylmethylamino)phenyl)-3-deutero-3-hydroxypropylcarbamate
following the method used in Example 20 gave Example 81 as a white
solid. Yield (0.14 g, quant); .sup.1H NMR (400 MHz, DMSO-d.sub.6+5%
D.sub.2O) .delta. 7.25 (t, J=8.0 Hz, 1H), 7.12 (br. s, 1H),
6.91-6.97 (m, 2H), 2.96 (d, J=6.8 Hz, 1H), 2.78-2.86 (m, 2H),
1.42-1.86 (m, 8H), 0.88-1.18 (m, 5H).
Example 82
Preparation of
3-amino-1-(3-(cyclohexylmethylamino)phenyl)-2,2-dideuteropropan-1-ol
##STR00382##
[1469]
3-Amino-1-(3-(cyclohexylmethylamino)phenyl)-2,2-dideuteropropan-1-o-
l was prepared following the method used in Examples 5 and 11.
[1470] Step 1: Addition of CD.sub.3CN to aldehyde 10 following the
method described in Example 5 gave
2,2-dideutero-3-hydroxy-3-(3-nitrophenyl)propanenitrile as a light
yellow solid. Yield (2.5 g, 39%); .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.34 (t, J=1.6 Hz, 1H), 8.16-8.19 (m, 1H), 7.82-7.84 (m,
1H), 7.62 (t, J=8.0 Hz, 1H), 5.10 (s, 1H).
[1471] Step 2: Hydrogenation of
2,2-dideutero-3-hydroxy-3-(3-nitrophenyl)propanenitrile with
aldehyde 29 following the method described in Example 11 gave
3-(3-(cyclohexylmethylamino)phenyl)-2,2-dideutero-3-hydroxypropanenitrile
as a colorless oil. Yield (0.46 g, 68%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.97 (t, J=8.0 Hz, 1H), 6.56 (t, J=1.2 Hz,
1H), 6.47 (d, J=7.6 Hz, 1H), 6.42 (dd, J=8.0, 1.6 Hz, 1H), 5.71 (d,
J=4.4 Hz, 1H), 5.57 (t, J=6.0 Hz, 1H), 4.68 (d, J=4.4 Hz, 1H), 2.80
(t, J=6.0 Hz, 2H), 1.44-1.78 (m, 6H), 1.08-1.21 (m, 3H), 0.84-0.96
(m, 2H).
[1472] Step 3: BH.sub.3-Me.sub.2S reduction of
3-(3-(cyclohexylmethylamino)phenyl)-2,2-dideutero-3-hydroxypropanenitrile
following the method described in Example 11 gave Example 82.
Example 83
Preparation of
3-amino-1-(3-(cyclohexylmethylamino)phenyl)-3,3-dideuteropropan-1-ol
##STR00383##
[1474]
3-Amino-1-(3-(cyclohexylmethylamino)phenyl)-3,3-dideuteropropan-1-o-
l was prepared following the method used in Example 20.
[1475] Step 1: LiAlD.sub.4 (0.012 g, 2.88 mmol) was added to a
solution of nitrile 30 (0.5 g, 1.92 mmol) in ether was added
LiAlD.sub.4 (0.012 g, 2.88 mmol) at 0.degree. C. The reaction
mixture was stirred at .degree. C. for 2 hr. The reaction was
quenched by slow addition of aqueous Na.sub.2SO.sub.4, the mixture
was then diluted with MTBE, dried over MgSO.sub.4 and concentrated
under reduced pressure. The residue was redissolved in DCM,
(Boc).sub.2O (0.6 g, 3.84 mmol) and Et.sub.3N (1.0 ml) were added.
The resulting mixture was stirred at room temperature for 18 hr,
concentrated under reduced pressure. Purification by flash
chromatography (30% to 50% EtOAc-hexanes gradient) gave tert-butyl
3-(tert-butoxycarbonyloxy)-3-(3-(cyclohexylmethylamino)phenyl)-1,1-dideut-
eropropylcarbamate as a colorless oil. Yield (0.23 g, 26%); .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 7.30 (d, J=8.0 Hz, 1H),
7.20-7.22 (m, 2H), 7.08 (d, J=7.6 Hz, 1H), 4.67 (t, J=6.4 Hz, 1H),
3.49 (d, J=7.2 Hz, 2H), 1.81 (d, J=6.4 Hz, 2H), 1.60-1.72 (m, 6H),
1.40-1.42 (m, 18H), 1.10-1.22 (m, 3H), 0.86-0.99 (m, 2H).
[1476] Step 2: Deprotection of tert-butyl
3-(tert-butoxycarbonyloxy)-3-(3-(cyclohexylmethylamino)phenyl)-1,1-didete-
ropropylcarbamate following the method used in Example 20 gave
Example 83 as a yellow solid. Yield (0.14 g, 90%); .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 7.48-7.60 (m, 3H), 7.38-7.42 (m, 1H), 4.91
(dd, J=9.2, 3.6 Hz, 1H), 3.24-3.33 (m, 2H), 1.66-2.08 (m, 8H),
1.24-1.36 (m, 3H), 0.96-1.06 (m, 2H).
Example 84
Preparation of
N-(3-(3-amino-3,3-dideutero-1-hydroxypropyl)phenyl)cyclohexanecarboxamide
##STR00384##
[1478]
N-(3-(3-Amino-3,3-dideutero-1-hydroxypropyl)phenyl)cyclohexanecarbo-
xamide is prepared following the method described below.
[1479] Step 1: Reduction of
3-(3-aminophenyl)-3-hydroxypropanenitrile (12) following the method
used in Example 83 gives
3-amino-1-(3-aminophenyl)-3,3-dideuteropropan-1-ol.
[1480] Step 2: Protection of
3-amino-1-(3-aminophenyl)-3,3-dideuteropropan-1-ol with Boc.sub.2O
following the method used in Example 15 gives tert-butyl
3-(3-aminophenyl)-1,1-dideutero-3-hydroxypropylcarbamate.
[1481] Step 3: Acylation of tert-butyl
3-(3-aminophenyl)-1,1-dideutero-3-hydroxypropylcarbamate by acyl
chloride 36 following the method used in Example 15 gives
tert-butyl
3-(3-(cyclohexanecarboxamido)phenyl)-1,1-dideutero-3-hydroxypropylcarbama-
te.
[1482] Step 4: Deprotection of tert-butyl
3-(3-(cyclohexanecarboxamido)phenyl)-1,1-dideutero-3-hydroxypropylcarbama-
te following the method used in Example 15 gives Example 84
hydrochloride.
Example 85
Preparation of
N-(3-(3-amino-3,3-dideutero-1-hydroxypropyl)phenyl)cyclohexanesulfonamide
##STR00385##
[1484]
N-(3-(3-Amino-3,3-dideutero-1-hydroxypropyl)phenyl)cyclohexanesulfo-
namide is prepared following the method used in Examples 84, 5, and
15.
[1485] Step 1: Sulfonation of tert-butyl
3-(3-aminophenyl)-1,1-dideutero-3-hydroxypropylcarbamate by
sulfonyl chloride 8 following the method used in Example 5 gives
tert-butyl
3-(3-(cyclohexanesulfonamido)phenyl)-1,1-dideutero-3-hydroxypropylcarbama-
te.
[1486] Step 2: Deprotection of tert-butyl
3-(3-(cyclohexanesulfonamido)phenyl)-1,1-dideutero-3-hydroxypropylcarbama-
te following the method used in Example 15 gives Example 84
hydrochloride.
Example 86
Preparation of
(R)-3-amino-1-(3-(cyclohexylmethylamino)phenyl)propan-1-ol
##STR00386##
[1488] (R)-3-Amino-1-(3-(cyclohexylmethylamino)phenyl)propan-1-ol
is prepared following the method described below.
[1489] Step 1: A mixture of aniline 33, Boc.sub.2O and 4-DMAP are
stirred under reflux until no starting aniline is seen by TLC. The
reaction mixture partitioned between aqueous NH.sub.4Cl and EtOAc
and aqueous layer additionally extracted with EtOAc. Organic layer
is then washed with brine, dried over anhydrous MgSO.sub.4 and
concentrated under reduced pressure. Purification by flash
chromatography (EtOAc-hexanes gradient) gives tert-butyl
3-(3-(tert-butoxycarbonyl(cyclohexylmethypamino)phenyl)-3-oxopropylcarbam-
ate.
[1490] Step 2: A mixture of tert-butyl
3-(3-(tert-butoxycarbonyl(cyclohexylmethyl)amino)phenyl)-3-oxopropylcarba-
mate and (+)-Ipc.sub.2BCl in anhydrous THF is stirred at room
temperature until no starting material is seen by TLC. The reaction
is then quenched with aqueous NH.sub.4Cl and stirred at room
temperature. Extraction with EtOAc and drying over anhydrous
MgSO.sub.4 followed by flash chromatography (EtOAc-hexanes
gradient) gives tert-butyl
(R)-3-(3-(tert-butoxycarbonyl(cyclohexylmethyl)amino)phenyl)-3-hydroxypro-
pylcarbamate.
[1491] Step 3: Deprotection of tert-butyl
(R)-3-(3-(tert-butoxycarbonyl(cyclohexylmethyl)amino)phenyl)-3-hydroxypro-
pylcarbamate following the method used in Example 12 gives Example
86 hydrochloride.
Example 87
Preparation of
3-amino-1-(3-(cyclohexylmethylamino)phenyl)-2-methylpropan-1-ol
##STR00387##
[1493]
3-Amino-1-(3-(cyclohexylmethylamino)phenyl)-2-methylpropan-1-ol is
prepared following the method used in Examples 5 and 11.
[1494] Step 1: Addition of propiononitrile to aldehyde 10 following
the method used in Example 5 gives
3-hydroxy-2-methyl-3-(3-nitrophenyl)propanenitrile.
[1495] Step 2: Hydrogenation of the mixture of
3-hydroxy-2-methyl-3-(3-nitrophenyl)propanenitrile and aldehyde 29
following the method used in Example 11 gives
3-(3-(cyclohexylmethylamino)phenyl)-3-hydroxy-2-methylpropanenitrile.
[1496] Step 3: BH.sub.3-Me.sub.2S reduction of
3-(3-(cyclohexylmethylamino)phenyl)-3-hydroxy-2-methylpropanenitrile
following the method used in Example 12 gives Example 87.
Example 88
Preparation of
1-amino-3-(3-(cyclohexylmethylamino)phenyl)propan-2-ol
##STR00388##
[1498] 1-Amino-3-(3-(cyclohexylmethylamino)phenyl)propan-2-ol is
prepared following the method described below.
[1499] Step 1: A mixture of Example 23 and Boc.sub.2O in
CH.sub.2Cl.sub.2 are stirred at room temperature until no starting
material is seen by TLC. The reaction mixture is then concentrated
under reduced pressure to give (E)-tert-butyl
3-(3-(cyclohexylmethylamino)phenyl)allylcarbamate.
[1500] Step 2: To a solution of (E)-tert-butyl
3-(3-(cyclohexylmethylamino)phenyl)allylcarbamate in
CH.sub.2Cl.sub.2 is added MCPBA (77%) followed by Na.sub.2CO.sub.3.
The reaction mixture is stirred at room temperature until no
starting material is seen by TLC. Aqueous NaHCO.sub.3 (10%) is
added and the product is extracted with CH.sub.2Cl.sub.2 three
times. Combined organic layers are washed with brine-NaHCO.sub.3,
dried over anhydrous Na.sub.2SO.sub.4 and concentrated under
reduced pressure. Purification by flash chromatography (10% to 50%
EtOAc-hexanes gradient) gives tert-butyl
(3-(3-(cyclohexylmethylamino)phenyl)oxiran-2-yl)methylcarbamate
which is used in the next step without further purification.
[1501] Step 3: A mixture of tert-butyl
(3-(3-(cyclohexylmethylamino)phenyl)oxiran-2-yl)methylcarbamate,
HCOOH.Et.sub.3N complex (5:2), Pd/C (10% wt) in absolute EtOH is
degassed by applying vacuum/argon 3 times. The reaction mixture is
stirred at room temperature until no starting material is seen by
TLC, then concentrated under reduced pressure. Purification by
flash chromatography (EtOAc-hexanes gradient) gives tert-butyl
3-(3-(cyclohexylmethylamino)phenyl)-2-hydroxypropylcarbamate.
[1502] Step 4: tert-Butyl
3-(3-(cyclohexylmethylamino)phenyl)-2-hydroxypropylcarbamate is
deprotected following the method used in Example 12 to give Example
88 hydrochloride.
Example 89
Preparation of
N-(3-(3-(cyclohexylmethylamino)phenyl)-3-hydroxypropyl)acetamide
##STR00389##
[1504]
N-(3-(3-(Cyclohexylmethylamino)phenyl)-3-hydroxypropyl)acetamide is
prepared following the method shown below.
[1505] Step 1: A mixture of Example 11 and 2,5-dioxopyrrolidin-1-yl
acetate in CH.sub.2Cl.sub.2 are stirred at room temperature until
no starting material is seen by TLC then concentrated under reduced
pressure. Purification by flash chromatography (EtOAc-hexanes
gradient) gives Example 89.
Example 90
Preparation of
3-amino-1-(3-((cyclohexylmethyl)(methyl)amino)phenyl)propan-1-ol
##STR00390##
[1507]
3-Amino-1-(3-((cyclohexylmethyl)(methyl)amino)phenyl)propan-1-ol
was prepared following the method described below.
[1508] Step 1: A mixture of aniline 32 (0.118 g, 0.327 mmol), DIPEA
(0.060 mL) and methyl iodide (0.094 g, 0.661 mmol) in absolute EtOH
was stirred at +75.degree. C. for 28 hrs. The reaction mixture was
concentrated under reduced pressure. Purification by column
chromatography (30% EtOAc-hexanes) gave tert-butyl
3-(3-((cyclohexylmethyl)(methyl)amino)phenyl)-3-hydroxypropylcarbamate
as a colorless oil. Yield (0.060 g, 49%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.16 (t, J=7.8 Hz, 1H), 6.52-6.68 (m, 3H), 4.92
(br. s, 1H), 4.68 (t, J=6.3 Hz, 1H), 3.36-3.50 (m, 1H), 3.14-3.23
(m, 1H), 3.11 (d, J=6.7 Hz, 2H), 2.94 (s, 3H), 1.87 (q, J=6.7 Hz,
2H), 1.58-1.76 (m, 6H), 1.38-1.49 (m, 10H), 1.08-1.28 (m, 3H),
0.86-1.00 (m, 2H).
[1509] Step 2: Deprotection of tert-butyl
3-(3-((cyclohexylmethyl)(methyl)amino)phenyl)-3-hydroxypropylcarbamate
following the method used in Example 11 gave Example 90
hydrochloride as a colorless oil. Yield (0.057 g, quant.); .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 7.75-7.79 (m, 1H), 7.54-7.63 (m,
3H), 4.94 (dd, J=3.5, 9.0 Hz, 1H), 3.40-3.60 (br. s, 1H), 3.20-3.30
(m, 2H), 3.05-3.18 (m, 5H), 2.02-2.14 (m, 1H), 1.91-2.02 (m, 1H),
1.56-1.74 (m, 4H), 1.27-1.40 (m, 1H), 0.95-1.22 (m, 5H); RP-HPLC
(Method 1) t.sub.R=5.10 min, 71.9% (AUC); ESI MS m/z 277.3
[M+H].sup.+.
Example 91
Preparation of
3-amino-1-(3-((1-deuterocyclohexyl)methylamino)phenyl)propan-1-ol
##STR00391##
[1511]
3-Amino-1-(3-((1-deuterocyclohexyl)methylamino)phenyl)propan-1-ol
was prepared following the method described below.
[1512] Step 1. To a solution of 1-deuteroclohexanecarboxylic acid
(5.0 g, 38.7 mmol) in anhydrous DMSO was added KOH (2.39 g, 42.6
mmol) with stirring for 5 min. Methyl iodide (6.59 g, 46.4 mmol)
was added and the reaction mixture was stirred overnight at room
temperature. Saturated NaHCO.sub.3 and ether was added and the
mixture was washed with brine, dried over Na.sub.2SO.sub.4 and
evaporated to dryness giving methyl 1-deuterocyclohexanecarboxylate
as a clear liquid. Yield (5.62 g, quant.); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 3.55 (s, 3H), 1.78-1.75 (m, 2H), 1.65-1.60
(m, 2H), 1.57-1.52 (m, 1H), 1.34-1.09 (m, 5H).
[1513] Step 2. To a solution of methyl
1-deuterocyclohexanecarboxylate (5.0 g, 34.9 mmol) in anhydrous
CH.sub.2Cl.sub.2 on an ice bath was added a solution of DIBAL-H in
CH.sub.2Cl.sub.2 (1.0 M, 73.3 ml, 73.3 mmol) The reaction mixture
was allowed to warm to room temperature over 2 hrs and quenched
with Rochelle's salt (100 ml). The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to give
(1-deuterocyclohexyl)methanol as a clear liquid. Yield (3.99 g,
97%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.27 (t, J=5.2
Hz, 1H), 3.15 (d, J=5.2 Hz, 2H), 1.66-1.56 (m, 5H), 1.21-1.20 (m,
3H), 0.84-0.78 (m, 2H).
[1514] Step 3. To a solution of (1-deuterocyclohexyl)methanol (3.0
g, 26.0 mmol) in anhydrous CH.sub.2Cl.sub.2 on an ice bath was
added Et.sub.3N (2.98 g, 28.6 mmol) and methanesulfonyl chloride
(3.28 g, 28.6 mmol). The reaction mixture was warmed to room temp
over 2 hr. 1N HCl was added and layers were separated. The organic
layer was dried over Na.sub.2SO.sub.4 and concentrated under
reduced pressure to give (1-deuterocyclohexyl)methyl
methanesulfonate as an off white solid. Yield (4.92 g, 98%);
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.97 (s, 2H), 3.12 (s,
3H), 1.68-1.58 (m, 5H), 1.25-1.08 (m, 3H), 0.97-0.88 (m, 2H).
[1515] Step 4: A mixture of aniline 12 (0.478 g, 2.95 mmol) and
(1-deuterocyclohexyl)methyl methanesulfonate (0.243 g, 1.26 mmol)
in absolute EtOH was stirred under argon at +70.degree. C. for 2
days. The reaction mixture was concentrated under reduced pressure.
Purification by flash chromatography (3% of 7N NH.sub.3/MeOH in
CH.sub.2Cl.sub.2) gave
3-(3-((1-deuterocyclohexyl)methylamino)phenyl)-3-hydroxypropanenitrile
as a yellow oil which crystallized on standing to off-white solid.
Yield (0.157 g, 48%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
6.97 (t, J=7.8 Hz, 1H), 6.54-6.59 (m, 1H), 6.45-6.50 (m, 1H),
6.40-6.45 (m, 1H), 5.72 (d, J=4.3 Hz, 1H), 5.56 (t, J=5.7 Hz, 1H),
4.66-4.72 (m, 1H), 2.80 (d, J=5.7 Hz, 2H), 2.65-2.80 (m, 2H),
1.54-1.78 (m, 5H), 1.05-1.22 (m, 3H), 0.83-0.97 (m, 2H).
[1516] Step 5: Reduction of
3-(3-((l-deuterocyclohexyl)methylamino)phenyl)-3-hydroxypropanenitrile
following the method used in Example 35 gave crude Example 91
hydrochloride as a colorless oil. This was partitioned between
CH.sub.2Cl.sub.2 and sat. NaHCO.sub.3, aqueous layer was extracted
with CH.sub.2Cl.sub.2. Combined organic layers were washed with
brine, concentrated under reduced pressure. Purification by flash
chromatography (4% to 10% of 7N
NH.sub.3/MeOH/CH.sub.2Cl.sub.2--CH.sub.2Cl.sub.2 gradient) gave
Example 91 as a colorless oil. Yield (0.0827 g, 23% over two
steps); .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.04 (t, J=7.8
Hz, 1H), 6.61-6.63 (m, 1H), 6.55-6.59 (m, 1H), 6.49 (ddd, J=0.8,
2.3, 8.0 Hz, 1H), 4.59 (dd, J=5.5, 7.8 Hz, 1H), 2.90 (s, 2H),
2.64-2.77 (m, 2H), 1.63-1.92 (m, 7H), 1.14-1.32 (m, 3H), 0.92-1.02
(m, 2H); RP-HPLC (Method 1) t.sub.R=5.20 min, 91.7% (AUC); ESI MS
m/z 264.3 [M+H].sup.+.
Example 92
Preparation of
3-amino-1-(3-(cyclohexyldideuteromethylamino)phenyl)propan-1-ol
##STR00392##
[1518]
3-Amino-1-(3-(cyclohexyldideuteromethylamino)phenyl)propan-1-ol is
prepared following the method described below.
[1519] Step 1. A solution of methyl cyclohexane carboxylate (9.99
g, 70.3 mmol) was added under inert atmosphere to a cooled
(0.degree. C.) suspension of LiAlD.sub.4 (2.99 g, 71.2 mmol) in
anhydrous Et.sub.2O. The reaction mixture was stirred at 0.degree.
C. for 3 hrs and then slowly quenched by addition of saturated
Na.sub.2SO.sub.4 until white precipitate formed. The mixture was
dried over anhydrous MgSO.sub.4, filtered. The filtrate was
concentrated under reduced pressure to give
cyclohexyldideuteromethanol as a colorless volatile liquid. Yield
(2.52 g, 32%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.63-1.78
(m, 5H), 1.40-1.50 (m, 1H), 1.10-1.35 (m, 4H), 0.86-0.99 (m,
2H).
[1520] Step 2. Mesylation of cyclohexyldideuteromethanol following
the method used in Example 91 gave cyclohexyldideuteromethyl
methanesulfonate as a colorless oil. Yield (4.14 g, 97%); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 2.98 (s, 3H), 1.64-1.80 (m, 6H),
1.10-1.32 (m, 3H), 0.92-1.05 (m, 2H).
[1521] Step 3: Alkylation of aniline 12 with
cyclohexyldideuteromethyl methanesulfonate following the method
used in Example 91 gave
3-(3-(cyclohexyldideuteromethylamino)phenyl)-3-hydroxypropanenitrile
as an off-white solid. Yield (0.128 g, 42%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.97 (t, J=7.8 Hz, 1H), 6.56 (t, J=1.8 Hz,
1H), 6.45-6.49 (m, 1H), 6.42 (ddd, J=0.8, 2.35, 8.0 Hz, 1H), 5.71
(d, J=4.3 Hz, 1H), 5.54 (br. s, 1H), 4.65-4.72 (m, 1H), 2.78 (ABd,
J=4.9, 16.6 Hz, 1H), 2.69 (ABd, J=6.65, 6.62 Hz, 1H), 1.70-1.79 (m,
2H), 1.54-1.70 (m, 3H), 1.48 (tt, J=3.5, 11.2 Hz, 1H), 1.07-1.22
(m, 3H), 0.84-0.95 (m, 2H).
[1522] Step 4: Reduction of
3-(3-(cyclohexyldideuteromethylamino)phenyl)-3-hydroxypropanenitrile
by BH.sub.3-Me.sub.2S following the method used in Example 91 gives
Example 92.
Example 93
Preparation of
N-(3-(3-amino-1-hydroxypropyl)phenyl)-1,2,2,3,3,4,4,5,5,6,6-undecadeutero-
cyclohexanecarboxamide
##STR00393##
[1524]
N-(3-(3-Amino-1-hydroxypropyl)phenyl)-1,2,2,3,3,4,4,5,5,6,6-undecad-
euterocyclohexanecarboxamide was prepared following the method
below.
[1525] Step 1: Oxalyl chloride (0.25 mL, 2.89 mmol) was added at
room temperature to a solution of perdeuterocyclohexanecarboxylic
acid (0.337 g, 2.42 mmol) in anhydrous CH.sub.2Cl.sub.2. DMF (0.05
mL) was then added and the reaction mixture was stirred at RT for 5
min, concentrated under reduced pressure and re-dissolved in
anhydrous CH.sub.2Cl.sub.2. This solution was then added to a
stirred solution of aniline 35 (0.36 g, 1.35 mmol) in anhydrous
CH.sub.2Cl.sub.2. After stirring overnight the mixture was
concentrated under reduced pressure. Purification by flash
chromatography (50% to 100% EtOAc-hexanes gradient) gave tert-butyl
(3-hydroxy-3-(3-(perdeuterocyclohexanecarboxamido)phenyl)propyl)carbamate
as a white solid. Yield (0.39 g, 75%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.71 (s, 1H), 7.52-7.56 (m, 1H), 7.41-7.46
(m, 1H), 7.17 (t, J=7.8 Hz, 1H), 6.90-6.94 (m, 1H), 6.73 (t, J=5.1
Hz, 1H), 5.15 (d, J=4.3 Hz, 1H), 4.46 (dt, J=6.5, 4.7 Hz, 1H),
2.90-2.98 (m, 2H), 1.60-1.68 (m, 2H), 1.34 (s, 9H).
[1526] Step 2: A mixture of tert-butyl
(3-hydroxy-3-(3-(perdeuterocyclohexanecarboxamido)phenyl)propyl)carbamate
(0,159 g, 0.41 mmol) and HCl/i-PrOH (5.5 M, 3 mL) in EtOAc was
stirred at room temperature for 22 hrs, then concentrated under
reduced pressure. Purification by flash chromatography (20% to 100%
of 20% 7N NH.sub.3/MeOH/CH.sub.2Cl.sub.2--CH.sub.2Cl.sub.2
gradient) gave Example 93 as a colorless oil. Yield (0.090 g, 64%);
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.54-7.57 (m, 1H), 7.45
(ddd, J=0.98, 1.96, 8.02 Hz, 1H), 7.25 (t, J=7.8 Hz, 1H), 7.05-7.10
(m, 1H), 4.70 (dd, J=5.3, 7.6 Hz, 1H), 2.68-2.81 (m, 2H), 1.77-1.94
(m, 2H); RP-HPLC (Method 1) t.sub.R=6.83 min, 95.7% (AUC); ESI MS
m/z 288.3 [M+H].sup.+.
Example 94
Preparation of
1-(3-(cyclohexylmethylamino)phenyl)-3-(methylamino)propan-1-ol
##STR00394##
[1528]
1-(3-(Cyclohexylmethylamino)phenyl)-3-(methylamino)propan-1-ol is
prepared following the method described below.
[1529] Step 1: A mixture of carbamate 32 and sodium
bis(2-methoxyethoxy)aluminumhydride in anhydrous THF is stirred
under an inert atmosphere until no starting material is seen by
TLC. The reaction mixture is then quenched by slow addition of 1N
NaOH and partitioned between aqueous NaHCO.sub.3 and
CH.sub.2Cl.sub.2. Organic layer is dried over anhydrous
Na.sub.2SO.sub.4 and concentrated under reduced pressure.
Purification by flash chromatography
(NH.sub.3/MeOH/CH.sub.2Cl.sub.2--CH.sub.2Cl.sub.2 gradient) gives
Example 94.
Example 95
Preparation of 3-(3-aminopropyl)-N-pentylaniline
##STR00395##
[1531] 3-(3-Aminopropyl)-N-pentylaniline is prepared following the
method used in Example 13.
[1532] Step 1: Hydrogenation of aniline 17 and pentanal gives
tert-butyl 3-(3-(pentylamino)phenyl)propylcarbamate.
[1533] Step 2: Deprotection of tert-butyl
3-(3-(pentylamino)phenyl)propylcarbamate following the method used
in Example 11 gives Example 95 hydrochloride.
Example 96
Preparation of N-(3-(3-aminopropyl)phenyl)pentanamide
##STR00396##
[1535] N-(3-(3-Aminopropyl)phenyl)pentanamide is prepared following
the method used in Example 15.
[1536] Step 1: Acylation of aniline 17 with pentanoyl chloride
gives tert-butyl 3-(3-pentanamidophenyl)propylcarbamate.
[1537] Step 2: Deprotection of tert-butyl
3-(3-pentanamidophenyl)propylcarbamate following the method used in
Example 15 gives Example 96 hydrochloride.
Example 97
Preparation of
N-(3-(3-amino-1-hydroxypropyl)phenyl)cyclopentanesulfonamide
##STR00397##
[1539] N-(3-(3-Amino-1-hydroxypropyl)phenyl)cyclopentanesulfonamide
was prepared following the method used in Example 19, 12.
[1540] Step 1: Sulfonation of aniline 35 by cyclopentanesulfonyl
chloride following the method used in Example 19 gave tert-butyl
3-(3-(cyclopentanesulfonamido)phenyl)-3-hydroxypropylcarbamate as a
yellow semi-solid. Yield (0.26 g, 36%)..sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.74 (s, 1H), 7.30 (s, 1H), 7.28 (d, J=7.6
Hz, 1H), 7.14-7.08 (m, 2H), 5.98 (bs, 1H), 4.85 (m, 1H), 3.54-3.48
(m, 1H), 2.88 (dd, J=5.2, 16.4 Hz, 1H), 2.78 (dd, J=5.2, 16.4 Hz,
1H), 1.90-1.77 (m, 4H), 1.64-1.62 (m, 2H), 1.53-1.44 (m, 2H).
[1541] Step 2: Deprotection of tert-butyl
3-(3-(cyclopentanesulfonamido)phenyl)-3-hydroxypropylcarbamate
following the method used in Example 12 gave Example 97
hydrochloride as a pale brown oil. Yield (0.14 g, 54%); .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 7.32 (s, 1H), 7.29 (t, J=8.0 Hz, 1H),
7.14 (d, J=8.0 Hz, 2H), 4.83-4.75 (t, J=6.4 Hz, 1H), 3.59-3.51 (m,
1H), 2.98-2.86 (m, 2H), 2.04-1.97 (m, 2H), 1.96-1.80 (m, 4H),
1.80-1.74 (m, 2H), 1.62-1.60 (m, 2H). RP-HPLC (Method 6)
t.sub.R=3.81 min, 90.65% (AUC); ESI MS m/z 299.32 [M+H].sup.+.
Example 98
Preparation of
N-(3-(3-aminopropanoyl)phenyl)cyclopentanesulfonamide
##STR00398##
[1543] N-(3-(3-Aminopropanoyl)phenyl)cyclopentanesulfonamide is
prepared following the method used in Example 97, 20.
[1544] Step 1: Oxidation of tert-butyl
3-(3-(cyclopentanesulfonamido)phenyl)-3-hydroxypropylcarbamate
following the method used in Example 20 gives tert-butyl
3-(3-(cyclopentanesulfonamido)phenyl)-3-oxopropylcarbamate.
[1545] Step 2: Deprotection of tert-butyl
3-(3-(cyclopentanesulfonamido)phenyl)-3-oxopropylcarbamate
following the method used in Example 20 gives Example 98
hydrochloride.
Example 99
Preparation of N-(3-(3-aminopropyl)phenyl)benzenesulfonamide
##STR00399##
[1547] N-(3-(3-Aminopropyl)phenyl)benzenesulfonamide was prepared
following the method described below.
[1548] Step 1: Sulfonation of
2-(3-(3-aminophenyl)propyl)isoindoline-1,3-dione by benzenesulfonyl
chloride following the method used in Example 6 gave
N-(3-(3-(1,3-dioxoisoindolin-2-yl)propyl)phenyl)benzenesulfonamide
as a yellow semi-solid. Yield (0.80 g, 62%)..sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 10.18 (s, 1H), 7.87-7.81 (m, 4H), 7.72 (d,
J=7.6 Hz, 2H), 7.58-7.49 (m, 3H), 7.11 (t, J=8.0, 1H), 6.92 (s,
1H), 6.88-6.85 (m, 2H), 3.52 (t, J=7.2 Hz, 2H), 2.50-2.49 (m, 2H),
1.81-1.74 (m, 2H).
[1549] Step 2: Deprotection of
N-(3-(3-(1,3-dioxoisoindolin-2-yl)propyl)phenyl)benzenesulfonamide
following the method used in Example 31 gave Example 99 as a white
solid. Yield (0.26 g, 42%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.23 (d, J=6.4 Hz, 2H), 7.55-7.46 (m, 3H), 7.03 (t, J=7.6
Hz, 1H), 6.86 (s, 1H), 6.82 (d, J=8.0 Hz, 1H), 6.73 (d, J=7.2 Hz,
1H), 2.56 (t, J=7.2 Hz, 2H), 2.46 (t, J=7.6 Hz, 2H), 1.60 (quintet,
J=7.6 Hz, 2H). RP-HPLC (Method 6) t.sub.R=4.32 min, 99.85% (AUC);
ESI MS m/z 291.19 [M+H].sup.+.
Example 100
Preparation of 3-amino-1-(3-(benzylamino)phenyl)propan-1-ol
##STR00400##
[1551] 3-Amino-1-(3-(benzylamino)phenyl)propan-1-ol was prepared
following the method used in Example 11.
[1552] Step 1: NaBH(OAc).sub.3 (7.84 g, 36.99 mmol) was added to a
solution of aniline 12 (2.0 g, 12.33 mmol) and benzaldehyde (1.3 g,
12.33 mmol) in DCM. The resulting mixture was stirred at RT for 5 h
and quenched with saturated aqueous NaHCO.sub.3. Organic layer was
washed with water followed by brine and dried over anhydrous
Na.sub.2SO.sub.4. Organic layer was concentrated under reduced
pressure. Purification by flash chromatography (40% to 50%
EtOAc-hexanes gradient) gave
3-(3-(benzylamino)phenyl)-3-hydroxypropanenitrile as a pale yellow
oil. Yield (2.61 g, 83%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.36-7.29 (m, 4H), 7.23-7.19 (m, 1H), 6.99 (t, J=8.0 Hz,
1H), 6.68 (s, 1H), 6.54 (d, J=7.6 Hz, 1H), 6.44 (dd, J=1.6, 8.0 Hz,
1H), 6.27 (t, J=6.0 Hz, 1H), 5.79 (d, J=4.4 Hz, 1H), 4.72-4.68 (m,
1H), 4.25 (d, J=6.0 Hz, 2H), 2.82-2.67 (m, 2H).
[1553] Step 2: BH.sub.3-Me.sub.2S reduction of
3-(3-(benzylamino)phenyl)-3-hydroxypropanenitrile following the
method used in Example 11 gave crude
3-amino-1-(3-(benzylamino)phenyl)propan-1-ol as an off-white
semi-solid which was used directly in next step. Yield (2.0 g,
75%).
[1554] Step 3: Protection of
3-amino-1-(3-(benzylamino)phenyl)propan-1-ol with Boc.sub.2O gave
tert-butyl
benzyl(3-(3-((tert-butoxycarbonyl)amino)-1-hydroxypropyl)phenyl)carbamate
as an off white semi-solid. Yield (2.7 g, 84%); .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 7.31-7.27 (m, 2H), 7.26-7.16 (m, 4H),
7.14 (s, 1H), 7.09 (d, J=7.6 Hz, 1H), 7.02 (d, J=8.0 Hz, 1H), 6.76
(t, J=5.2 Hz, 1H), 5.20 (d, J=4.4 Hz, 1H), 4.81 (d, J=6.0 Hz, 2H),
4.52-4.47 (m, 1H), 2.93 (q, J=6.4 Hz, 2H), 1.65-1.60 (m, 2H), 1.36
(s, 18H).
[1555] Step 4: Deprotection of tert-butyl
benzyl(3-(3-((tert-butoxycarbonyl)amino)-1-hydroxypropyl)phenyl)carbamate
gave Example 100 hydrochloride as a yellow solid. Yield (0.5 g,
69%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.06 (bs, 3H),
7.45 (d, J=6.0 Hz, 2H), 7.36-7.24 (m, 4H), 7.19 (bs, 1H), 7.05 (bs,
2H), 4.64-4.62 (m, 1H), 4.42 (s, 2H), 2.82-2.77 (m, 2H), 1.86-1.77
(m, 2H); RP-HPLC (Method 6) t.sub.R=4.48 min, 90.84% (AUC); ESI MS
m/z 257.22 [M+H].sup.+.
Example 101
Preparation of
N-(3-(3-amino-1-hydroxypropyl)phenyl)benzenesulfonamide
##STR00401##
[1557] N-(3-(3-Amino-1-hydroxypropyl)phenyl)benzenesulfonamide was
prepared following the method used in Example 5.
[1558] Step 1: Sulfonation of aniline 12 by benzenesulfonyl
chloride gave
N-(3-(2-cyano-1-hydroxyethyl)phenyl)benzenesulfonamide as a yellow
semi-solid. Yield (0.9 g, 81%);.sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 10.30 (s, 1H), 7.77-7.69 (m, 2H), 7.61-7.57 (m, 1H),
7.54-7.45 (m, 2H), 7.21 (d, J=4.8 Hz, 1H), 7.18 (d, J=8.0 Hz, 1H),
7.08-6.98 (m, 1H), 6.84 (d, J=8.0 Hz, 1H), 6.09 (d, J=3.6 Hz, 1H),
4.91-4.77 (m, 1H), 2.80 (dd, J=4.8, 16.8 Hz, 1H), 2.70 (dd, J=4.8,
16.8 Hz, 1H).
[1559] Step 2: BH.sub.3-Me.sub.2S reduction of
N-(3-(2-cyano-1-hydroxyethyl)phenyl)benzenesulfonamide gives
Example 101 as a white solid. Yield (0.385 g, 48%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.71 (dd, J=2.0, 7.2 Hz, 2H),
7.52-7.43 (m, 3H), 7.04 (t, J=8.0 Hz, 1H), 6.98 (s, 1H), 6.81 (t,
J=8.4 Hz, 2H), 4.48 (t, J=6.4 Hz, 1H), 2.60 (t, J=7.2 Hz, 2H), 1.61
(quintet, J=7.2 Hz, 2H); RP-HPLC (Method 6) t.sub.R=4.02 min,
94.69% (AUC); ESI MS m/z 307.29 [M+H].sup.+.
Example 102
Preparation of 3-amino-1-(3-(benzylamino)phenyl)propan-1-one
##STR00402##
[1561] 3-Amino-1-(3-(benzylamino)phenyl)propan-1-one was prepared
following the method used in Example 100 and 12.
[1562] Step 1: Oxidation of tert-butyl
benzyl(3-(3-((tert-butoxycarbonyl)amino)-1-hydroxypropyl)phenyl)carbamate
with Des-Martin periodinane following the method used in Example 40
gave tert-butyl
benzyl(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)carbamate
as a yellow oil. Yield (1.2 g, 74%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.75 (s, 1H), 7.71 (d, J=6.0 Hz, 1H),
7.45-7.41 (m, 2H), 7.32-7.29 (m, 2H), 7.23-7.19 (m, 3H), 6.80 (bs,
1H), 4.89 (s, 2H), 3.24 (m, 2H), 3.15-3.09 (m, 2H), 1.38 (s, 9H),
1.35 (s, 9H).
[1563] Step 2: Deprotection of tert-butyl
benzyl(3-(3-((tert-butoxycarbonyl)amino)propanoyl)phenyl)carbamate
gave Example 102 hydrochloride as a yellow oil. Yield (0.8 g,
yellow solid, 92%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
8.06 (bs, 3H), 7.45 (d, J=6.4 Hz, 2H), 7.45-7.22 (m, 7H), 7.01 (bs,
1H), 4.42 (s, 2H), 3.49-3.42 (m, 2H), 3.10-3.02 (m, 2H); RP-HPLC
(Method 6) t.sub.R=4.72 min, 73.30% (AUC); ESI MS m/z 255.20
[M+H].sup.+.
Example 103
Preparation of N-(3-(3-aminopropanoyl)phenyl)benzenesulfonamide
##STR00403##
[1565] N-(3-(3-Aminopropanoyl)phenyl)benzenesulfonamide was
prepared following the method used in Example 40.
[1566] Step 1: Protection of Example 101 following the method used
in Example 40 gave tert-butyl
tert-butoxycarbonyl(3-hydroxy-3-(3-(phenylsulfonamido)phenyl)propyl)carba-
mate as a colorless oil. Yield (0.36 g, 87%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.98 (dd, J=1.2, 8.0 Hz, 2H), 7.66 (t, J=7.6
Hz, 1H), 7.56 (t, J=8.0 Hz, 2H), 7.42-7.38 (m, 2H), 7.29 (s, 1H),
7.16 (d, J=6.8 Hz, 1H), 4.89 (bs, 1H), 4.80-4.78 (m, 1H), 3.52 (m,
1H), 3.45 (bs, 1H), 3.21-3.13 (m, 1H), 1.91-1.81 (m, 2H), 1.39 (s,
9H), 1.33 (s, 9H).
[1567] Step 2: Oxidation of tert-butyl
tert-butoxycarbonyl(3-hydroxy-3-(3-(phenylsulfonamido)phenyl)propyl)carba-
mate gives tert-butyl
tert-butoxycarbonyl(3-oxo-3-(3-(phenylsulfonamido)phenyl)propyl)carbamate
as a colorless oil. Yield (0.19 g, 76%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.06 (d, J=7.2 Hz, 1H), 7.99 (d, J=7.6 Hz,
2H), 7.84-7.79 (m, 2H), 7.72 (t, J=7.6 Hz, 2H), 7.68-7.62 (m, 2H),
6.82 (m, 1H), 3.31 (m, 2H), 3.21-3.18 (m, 2H), 1.36 (s, 9H), 1.24
(s, 9H).
[1568] Step 3: Deprotection of tert-butyl
3-oxo-3-(3-(phenylsulfonamido)phenyl)propylcarbamate gives Example
103 hydrochloride as a pale yellow solid. Yield (0.12 g, 93%);
.sup.1H NMR (400 MHz, MeOD) .delta. 7.82 (s, 1H), 7.79 (d, J=7.2
Hz, 2H), 7.72 (d, J=7.6 Hz, 1H), 7.56 (t, J=7.2 Hz, 1H), 7.47 (t,
J=7.6 Hz, 2H), 7.39 (t, J=8.0 Hz, 1H), 7.33 (d, J=8.0 Hz, 1H), 3.38
(t, J=6.0 Hz, 2H), 3.31 (m, 2H). RP-HPLC (Method 6)
t.sub.R.sup.=4.11 min, 98.36% (AUC); ESI MS m/z 305.25
[M+H].sup.+.
Example 104
Preparation of 3-(3-aminopropyl)-N-(2-methoxybenzyl)aniline
##STR00404##
[1570] 3-(3-Aminopropyl)-N-(2-methoxybenzyl)aniline was prepared
following the method used in Example 64.
[1571] Step 1: Hydrogenation of
2,2,2-trifluoro-N-(3-(3-nitrophenyl)allyl)acetamide and
2-methoxybenzaldehyde gave tert-butyl
3-(3-(2-methoxybenzylamino)phenyl)propylcarbamate as a colorless
semi-solid. Yield (0.16 g, 16%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.42 (br.s, 1H), 7.23-7.18 (m, 2H), 6.97 (d,
J=8.4 Hz, 1H), 6.93 (t, J=7.6 Hz, 1H), 6.86 (t, J=7.6 Hz, 1H), 6.40
(s, 1H), 6.35-6.34 (m, 2H), 5.94 (t, J=6.0 Hz, 1H), 4.18 (d, J=6.0
Hz, 2H), 3.82 (s, 3H), 3.16 (q, J=6.4, 2H), 2.41 (t, J=7.6 Hz, 2H),
1.70 (quintet, J=7.6, 2H).
[1572] Step 2: Deprotection of tert-butyl
3-(3-(2-methoxybenzylamino)phenyl)propylcarbamate following the
method used in Example 95 gave Example 104 as a light green
semi-solid. Yield (0.09 g, 76%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.23-7.19 (m, 2H), 6.98 (d, J=8.0 Hz, 1H),
6.93 (t, J=7.6 Hz, 1H), 6.87 (t, J=7.6 Hz, 1H), 6.39 (s, 1H), 6.34
(d, J=8.0 Hz, 2H), 5.95 (t, J=6.0 Hz, 1H), 4.18 (d, J=6.0 Hz, 2H),
3.83 (s, 3H), 2.62 (t, J=7.6 Hz, 2H), 2.44 (t, J=7.6 Hz, 2H), 1.65
(quintet, J=7.2 Hz, 2H); RP-HPLC (Method 6) t.sub.R=4.92 min,
97.97% (AUC); ESI MS m/z 271.28 [M+H].sup.+.
Example 105
Preparation of 3-(3-aminopropyl)-N-phenethylaniline
##STR00405##
[1574] 3-(3-Aminopropyl)-N-phenethylaniline was prepared following
the method used in Example 33 and 11.
[1575] Step 1: Hydrogenation of
(E)-2,2,2-trifluoro-N-(3-(3-nitrophenyl)allyl)acetamide and
2-phenylacetaldehyde gave
2,2,2-trifluoro-N-(3-(3-(phenethylamino)phenyl)propyl)acetamide as
a colorless semi-solid. Yield (0.3 g, 24%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.43 (bs, 1H), 7.31-7.18 (m, 5H) 6.95 (t,
J=7.6 Hz, 1H), 6.49-6.36 (m, 3H), 5.56 (t, J=6.0 Hz, 1H), 3.24-3.14
(m, 4H), 2.87-2.82 (m 2H), 2.37 (t, J=7.6 Hz, 2H), 1.74 (quintet,
J=7.6 Hz, 2H).
[1576] Step 2: Deprotection of
2,2,2-trifluoro-N-(3-(3-(phenethylamino)phenyl)propyl)acetamide
following the method used in Example 23 gave Example 105 as a light
green semi-solid. Yield (0.12 g, 55%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.30-7.18 (m, 5H), 6.98 (t, J=8.0 Hz, 1H),
6.41 (d, J=7.2 Hz, 2H), 6.37 (d, J=7.6 Hz, 1H), 5.57 (t, J=5.6 Hz,
1H), 3.21 (q, J=6.4, Hz, 2H), 2.82 (t, J=7.6 Hz, 2H), 2.64 (t,
J=7.6 Hz, 2H), 2.50-2.46 (m, 2H), 1.69 (quintet, J=7.6, 2H);
RP-HPLC (Method 6) t.sub.R=5.13 min, 97.42% (AUC); ESI MS m/z
255.24 [M+H].sup.+.
Example 106
Preparation of 3-(3-aminopropyl)-N-(thiazol-2-ylmethyl)aniline
##STR00406##
[1578] 3-(3-Aminopropyl)-N-(thiazol-2-ylmethyl)aniline was prepared
following the method described below.
[1579] Step 1: .ANG.-3 Molecular seives were added to a solution of
aniline 17 (0.4 g, 1.6 mmol) and thiazole-2-carbaldehyde (0.18 g,
1.6 mmol) in MeOH. The reaction mixture was stirred for 18 h and
then NaBH.sub.4 (0.121 g, 3.2 mmol) was added and the reaction
mixture was stirred for overnight. The reaction mixture was
filtered through Celite, concentrated under reduced pressure
Purification by column chromatography (100-200 silica mesh, 20%
EtOAc in hexane) gave tert-butyl
3-(3-(thiazol-2-ylmethylamino)phenyl)propylcarbamate as a brown
oil. Yield (0.17 g, 31%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.73 (d, J=3.2 Hz, 1H), 7.56 (d, J=3.2 Hz, 1H), 6.96 (t,
J=7.6 Hz, 1H), 6.80 (bs, 1H), 6.45-6.38 (m, 4H), 4.55 (d, J=6.0 Hz,
2H), 2.91 (q, J=6.4 Hz, 2H), 2.39 (t, J=7.2 Hz, 2H), 1.59 (quintet,
J=7.2 Hz, 2H), 1.37 (s, 9H).
[1580] Step 2: Deprotection of tert-butyl
3-(3-(thiazol-2-ylmethylamino)phenyl)propylcarbamate following the
method used in Example 11 gave Example 106 hydrochloride as a pale
brown solid. Yield (0.09 g, 31%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.90 (m, 3H), 7.76 (d, J=3.2 Hz, 1H), 7.61
(d, J=3.2 Hz, 1H), 7.00 (t, J=7.6 Hz, 1H), 6.48-6.45 (m, 3H), 4.55
(s, 2H), 4.77 (bs, 1H), 2.73 (t, J=6.4 Hz, 2H), 2.50 (m, 2H), 1.77
(quintet, J=7.6 Hz, 2H); RP-HPLC (Method 6) t.sub.R=3.92 min,
99.76% (AUC); ESI MS m/z 248.20 [M+H].sup.+.
Example 107
Preparation of
N-(3-(3-aminopropyl)phenyl)-2-cyclohexylethanesulfonamide
##STR00407##
[1582] N-(3-(3-Aminopropyl)phenyl)-2-cyclohexylethanesulfonamide is
prepared following the method used in Example 6.
[1583] Step 1: Sulfonation of aniline 17 by
2-cyclohexylethanesulfonyl chloride following the method used in
Example 6 gives tert-butyl
3-(3-(2-cyclohexylethylsulfonamido)phenyl)propylcarbamate.
[1584] Step 2: Deprotection of tert-butyl
3-(3-(2-cyclohexylethylsulfonamido)phenyl)propylcarbamate gives
Example 107 hydrochloride.
Example 108
Preparation of
N-(3-(3-aminopropanoyl)phenyl)-2-cyclohexylethanesulfonamide
##STR00408##
[1586] N-(3-(3-Aminopropanoyl)phenyl)-2-cyclohexylethanesulfonamide
is prepared following the method used in Example 97.
[1587] Step 1: Sulfonation of aniline 35 by
2-cyclohexylethanesulfonyl chloride gives tert-butyl
3-(3-(2-cyclohexylethylsulfonamido)phenyl)-3-hydroxypropylcarbamate.
[1588] Step 2: Deprotection of tert-butyl
3-(3-(2-cyclohexylethylsulfonamido)phenyl)-3-hydroxypropylcarbamate
gives Example 108 hydrochloride.
Example 109
Preparation of
N-(3-(3-amino-1-hydroxypropyl)phenyl)-2-cyclohexylethanesulfonamide
##STR00409##
[1590]
N-(3-(3-Amino-1-hydroxypropyl)phenyl)-2-cyclohexylethanesulfonamide
is prepared following the method used in Example 98.
[1591] Step 1: Oxidation of tert-butyl
3-(3-(2-cyclohexylethylsulfonamido)phenyl)-3-hydroxypropylcarbamate
gives tert-butyl
3-(3-(2-cyclohexylethylsulfonamido)phenyl)-3-oxopropylcarbamate.
[1592] Step 2: Deprotection of tert-butyl
3-(3-(2-cyclohexylethylsulfonamido)phenyl)-3-oxopropylcarbamate
gives Example 109 hydrochloride.
Example 110
Preparation of 3-(3-aminopropyl)-N-(5-(benzyloxy)pentyl)aniline
##STR00410##
[1594] 3-(3-Aminopropyl)-N-(5-(benzyloxy)pentyl)aniline is prepared
following the method used in Example 95.
[1595] Step 1: Hydrogenation of aniline 17 and
5-(benzyloxy)pentanal gives tert-butyl
34345-(benzyloxy)pentylamino)phenyl)propylcarbamate.
[1596] Step 2: Deprotection of tert-butyl
3-(3-(5-(benzyloxy)pentylamino)phenyl)propylcarbamate following the
method used in Example 11 gives Example 110 hydrochloride.
Example 111
Preparation of
N-(3-(3-aminopropyl)phenyl)-5-methoxypentane-1-sulfonamide
##STR00411##
[1598] N-(3-(3-Aminopropyl)phenyl)-5-methoxypentane-1-sulfonamide
is prepared following the method used in Example 6.
[1599] Step 1: Sulfonation of aniline 17 by hexane-1-sulfonyl
chloride following the method used in Example 6 gives tert-butyl
3-(3-(hexylsulfonamido)phenyl)propylcarbamate.
[1600] Step 2: Deprotection of tert-butyl
3-(3-(hexylsulfonamido)phenyl)propylcarbamate gives Example 111
hydrochloride.
Example 112
Preparation of
N-(3-(3-amino-1-hydroxypropyl)phenyl)-5-methoxypentane-1-sulfonamide
##STR00412##
[1602]
N-(3-(3-Amino-1-hydroxypropyl)phenyl)-5-methoxypentane-1-sulfonamid-
e is prepared following the method used in Example 97.
[1603] Step 1: Sulfonation of aniline 35 by
5-methoxypentane-1-sulfonyl chloride gives tert-butyl
3-hydroxy-3-(3-(5-methoxypentylsulfonamido)phenyl)propylcarbamate.
[1604] Step 2: Deprotection of tert-butyl
3-hydroxy-3-(3-(5-methoxypentylsulfonamido)phenyl)propylcarbamate
gives Example 112 hydrochloride.
Example 113
Preparation of
N-(3-(3-aminopropanoyl)phenyl)-5-methoxypentane-1-sulfonamide
##STR00413##
[1606]
N-(3-(3-Aminopropanoyl)phenyl)-5-methoxypentane-1-sulfonamide is
prepared following the method used in Example 98.
[1607] Step 1: Oxidation of tert-butyl
3-hydroxy-3-(3-(5-methoxypentylsulfonamido)phenyl)propylcarbamate
gives tert-butyl
3-(3-(5-methoxypentylsulfonamido)phenyl)-3-oxopropylcarbamate.
[1608] Step 2: Deprotection of tert-butyl
3-(3-(5-methoxypentylsulfonamido)phenyl)-3-oxopropylcarbamate gives
Example 113 hydrochloride.
Example 114
Preparation of
(E)-1-(3-(3-amino-1-deutero-1-hydroxypropyl)styryl)cyclohexanol
##STR00414##
[1610]
(E)-1-(3-(3-Amino-1-deutero-1-hydroxypropyl)styryl)cyclohexanol was
prepared following the method shown in Scheme 13.
##STR00415##
[1611] Step 1: To a cold (-50.degree. C.) solution of
t-BuO.sup.-K.sup.+ in THF (1M, 0.76 L, 760 mmol) under N.sub.2 was
slowly added acetonitrile (37.0 mL, 703 mmol). The reaction mixture
was stirred for 25 min and then a solution of 3-bromobenzaldehyde
(13.1) (75 mL, 640 mmol) in anhydrous THF was added dropwise
keeping the temperature below -40.degree. C. After addition was
complete, the reaction mixture as stirred at for 45 min while
slowly warming to -10.degree. C. The reaction mixture was
partitioned between THF and an aqueous solution of NH.sub.4Cl
(25%), organic layer was washed with brine, dried over anhydrous
MgSO.sub.4 and filtered. The filtrate was concentrated under
reduced pressure to give hydroxynitrile 13.2 as an amber oil. Yield
(148 g, quant.); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.60
(t, J=1.6 Hz, 1H), 7.46 (ddd, J=7.6, 2.0, 1.2 Hz, 1H), 7.40 (dd,
J=7.6, 2.0 Hz, 1H), 7.31 (t, J=7.6 Hz, 1H), 6.05 (d, J=4.8 Hz, 1H),
4.87-4.92 (m, 1H), 2.94-2.80 (m, 2H).
[1612] Step 2: To an ice cold solution of
3-(3-bromophenyl)-3-hydroxypropanenitrile (13.2) (2.70 g, 11.9
mmol) in anhydrous THF under argon was added a solution of
LiAlH.sub.4 in THF (11.9 mL of a 2 M solution in THF, 23.8 mmol).
The mixture was stirred at 0.degree. C. for 45 min, diluted with
ether (50 mL), and quenched with the dropwise addition of saturated
aqueous Na.sub.2SO.sub.4 (approximately 2 mL). After drying over
MgSO.sub.4, the mixture was filtered and concentrated under reduced
pressure to give amine 13.3 as a light green oil. This material was
used in the next step without further purification. Yield (2.30 g,
84%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.49 (m, 1H),
7.37 (dt, J=7.2, 1.6 Hz, 1H), 7.23-7.31 (m, 2H), 4.66 (t, J=6.8 Hz,
1H), 2.61 (m, 2H), 1.61 (q, J=6.8 Hz, 2H).
[1613] Step 3: To a solution of amine 13.3 (5.67 g, 24.6 mmol) in
anhydrous CH.sub.2Cl.sub.2 was added Boc.sub.2O (5.69 g, 26.1
mmol). The reaction mixture was stirred at room temperature for 15
min, concentrated under reduced pressure, the residue was dissolved
in CH.sub.2Cl.sub.2 and Celite (8.67 g) followed by pyridinium
chlorochromate (7.67 g, 35.6 mmol) was added. The reaction mixture
was stirred at room temperature for 17 hrs and solvent was removed
under reduced pressure. Dark brown residue was suspended in
EtOAc-hexanes (30%), filtered and the filtrate was concentrated
under reduced pressure. Purification by flash chromatography (20%
to 80% EtOAc-hexanes gradient) gave ketone 13.4 as a light yellow
oil. Yield (7.2 g, 89%); 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
8.0-8.04 (m, 1H), 7.87-7.93 (m, 1H), 7.78-7.83)m, 1H), 7.47 (t,
J=7.8 Hz, 1H), 6.78 (br. t, J=5.1 Hz, 1H), 3.25 (q, J=5.7 Hz, 2H),
3.12 (t, J=6.3 Hz, 2H), 1.33 (s, 9H).
[1614] Step 4: NaBD.sub.4 (1.07 g, 25.5 mmol) was added to stirred
solution of ketone 13.4 (3.30 g, 10.1 mmol) in i-PrOH. The reaction
mixture was stirred at room temperature for 30 min, aqueous
NH.sub.4Cl (25%) was carefully added. The product was extracted
with EtOAc, organic layer was washed with brine, dried over
anhydrous MgSO.sub.4 and concentrated under reduced pressure to
give 3-amino-1-(3-bromophenyl)-1-deuteropropan-1-ol as a colorless
oil. Yield (3.44 g, quant.); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.49 (t, J=1.6 Hz, 1H), 7.39 (dt, J=1.6, 7.4 Hz, 1H),
7.23-7.32 (m, 2H), 6.75 (br. t, J=4.9 Hz, 1H), 5.30 (s, 1H),
2.87-3.00 (m, 2H), 1.65 (t, J=7.0 Hz, 2H), 1.35 (s, 9H). A mixture
of 3-amino-1-(3-bromophenyl)-1-deuteropropan-1-ol (3.44 g),
HCl/i-PrOH (5.5 M, 30 mL) and Et.sub.2O was stirred at room
temperature for 6 hrs and concentrated under reduced pressure to
give amine hydrochloride 13.5 as a colorless oil. Yield (3.07 g,
quant.). The product was used in the next step without
purification.
[1615] Step 5: To a solution of salt 13.5 (3.07 g) in
CH.sub.2Cl.sub.2-MeOH (2:1) was added Et.sub.3N (1.8 mL, 12.9 mmol)
followed by CF.sub.3COOEt (3.0 mL, 25.1 mmol) and the reaction
mixture was stirred at room temperature overnight. The reaction
mixture was concentrated under reduced pressure, the residue was
partitioned between aq. NH.sub.4Cl (25%) and EtOAc. The aqueous
layer was extracted with EtOAc, combined organic layers were washed
with brine, dried over anhydrous MgSO.sub.4, and concentrated under
reduced pressure to give amide 13.6 as a light yellow oil. Yield
(3.14 g, 83%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.32
(br. s, 1H), 7.51 (t, J=1.8 Hz, 1H), 7.38-7.42 (m, 1H), 7.23-7.33
(m, 2H), 5.43 (s, 1H), 3.16-3.29 (m, 2H), 1.70-1.85 (m, 2H).
[1616] Step 6. Tetrabutylammonium acetate (2.0 g) was added to
N-(3-(3-bromophenyl)-3-deutero-3-hydroxypropyl)-2,2,2-trifluoroacetamide
(13.6) (0.72 g, 2.2 mmol), 1-vinylcyclohexanol (13.7) (0.416 g, 3.3
mmol) and Pd(OAc).sub.2 (0.01 g, 0.045 mmol). This mixture was
stirred under an atmosphere of argon at 90.degree. C. overnight.
H.sub.2O and EtOAc were added to the reaction mixture and layers
were separated. The organic layer was dried over Na.sub.2SO.sub.4
and concentrated under reduced pressure. Flash chromatography (30%
EtOAc/hexanes) gave alkene 13.8 as a light brown oil. Yield (0.53
g, 64%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.31 (t, J=5.0
Hz, 1H), 7.35 (s, 1H), 7.26-7.22 (m, 2H), 7.16-7.13 (m, 1H), 6.51
(d, J=16.0 Hz, 1H), 6.35 (d, J=16.0 Hz, 1H), 5.27 (br.s, 1H), 4.39
(br.s, 1H), 3.26-3.21 (m, 2H), 1.80-1.77 (m, 2H), 1.63-1.39 (m,
9H), 1.26-1.17 (m, 1H).
[1617] Step 7. To a solution of
(E)-N-(3-deutero-3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)prop-
yl)-2,2,2-trifluoroacetamide (13.8) (0.26 g, 0.69 mmol) in
H.sub.2O/MeOH (1:4) was added K.sub.2CO.sub.3 (0.48 g, 3.5 mmol).
This mixture was stirred at 50.degree. C. for 3 h and then
evaporated to near dryness. H.sub.2O and EtOAc were added to the
residue and the layers were separated. The organic layer was dried
over Na.sub.2SO.sub.4 and concentrated under reduced pressure.
Flash chromatography (10% MeOH/CH.sub.2Cl.sub.2) followed by (10%
7N NH.sub.3 in MeOH/CH.sub.2Cl.sub.2) gave Example 115 as a clear
oil. Yield (0.122 g, 64%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.35 (s, 1H), 7.29-7.15 (m, 2H), 7.14-7.11 (m, 1H), 6.50
(d, J=16.0 Hz, 1H), 6.33 (d, J=16.0 Hz, 1H), 4.40 (br.s, 1H),
2.66-2.53 (m, 2H), 1.66-1.49 (m, 4H), 1.47-1.39 (m, 7H), 1.25-1.17
(m, 1H); ESI MS m/z 277.3 [M+H].sup.+.
Example 115
Preparation of
(E)-3-amino-1-(3-(2-cyclohexylvinyl)phenyl)-2,2-dideuteropropan-1-ol
##STR00416##
[1619]
(E)-3-Amino-1-(3-(2-cyclohexylvinyl)phenyl)-2,2-dideuteropropan-1-o-
l was prepared following the method used in Example 114.
[1620] Step 1: Addition of trideuteroacetonitrile to
3-bromobenzaldehyde gave
3-(3-bromophenyl)-2,2-dideutero-3-hydroxypropanenitrile as a
colorless oil. Yield (5.17 g, 95%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.60 (t, J=1.6 Hz, 1H), 7.46 (ddd, J=1.2,
2.0, 7.8 Hz, 1H), 7.37-7.41 (m, 1H), 7.31 (t, J=7.6 Hz, 1H), 6.05
(d, J=4.0 Hz, 1H), 4.88 (m, 1H).
[1621] Step 2: A mixture of
3-(3-bromophenyl)-2,2-dideutero-3-hydroxypropanenitrile (1.91 g,
8.37 mmol), borane-dimethylsulfide (2.0 mL, 21.1 mmol) in anhydrous
THF was stirred under reflux for 15 hr. After cooling to room
temperature MeOH was carefully added to the reaction mixture
followed by HCl/MeOH (1.25 M, 10 mL). The mixture was stirred under
reflux for 4 hrs and concentrated under reduced pressure to give
3-amino-1-(3-bromophenyl)-2,2-dideuteropropan-1-ol hydrochloride as
a white foam which was used in the next step without purification.
Yield (2.25 g, quant.).
[1622] Step 3: To a solution of
3-amino-1-(3-bromophenyl)-2,2-dideuteropropan-1-ol hydrochloride
(2.25 g, 8.38 mmol) in CH.sub.2Cl.sub.2-MeOH (2:1) was added
CF.sub.3COOEt (3.0 mL) followed by Et.sub.3N (2.0 mL, 14.3 mmol).
The reaction mixture was stirred at room temperature for 1 h and
concentrated under reduced pressure. The residue was suspended in
EtOAc, washed with brine, dried over anhydrous MgSO.sub.4 and
concentrated under reduced pressure to give
N-(3-(3-bromophenyl)-2,2-dideutero-3-hydroxypropyl)-2,2,2-trifluoroacetam-
ide as a colorless oil. Yield (2.81 g, quant.); .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.32 (br.s, 1H), 7.49-7.52 (m, 1H), 7.40
(dt, J=1.6, 7.4 Hz, 1H), 7.24-7.32 (m, 2H), 5.44 (d, J=4.7 Hz, 1H),
4.56 (d, J=4.7 Hz, 1H), 3.16-3.27 (m, 2H).
[1623] Step 4. Heck coupling between
N-(3-(3-bromophenyl)-2,2-dideutero-3-hydroxypropyl)-2,2,2-trifluoroacetam-
ide and 1-vinylcyclohexanol following the method used in Example
114 gave
(E)-N-(2,2-dideutero-3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-
propyl)-2,2,2-trifluoroacetamide as a clear oil. Yield (0.32 g,
56%); .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.40 (s, 1H),
7.24-7.28 (m, 2H), 7.16-7.20 (m, 1H), 6.60 (d, J=16.4 Hz, 1H), 6.36
(d, J=16.4 Hz, 1H), 4.67 (s, 1H), 3.35 (s, 2H), 1.49-1.76 (m, 9H),
1.28-1.40 (m, 1H).
[1624] Step 5.
(E)-N-(2,2-dideutero-3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-
propyl)-2,2,2-trifluoroacetamide was deprotected following the
method used in Example 114 to give Example 115 as a light yellow
oil. Yield (0.22 g, quant.); .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.40 (s, 1H), 7.24-7.28 (m, 2H), 7.16-7.20 (m, 1H), 6.60
(d, J=16.4 Hz, 1H), 6.35 (d, J=16.4 Hz, 1H), 4.70 (s, 1H), 2.71 (d,
J=6.0 Hz, 2H), 1.49-1.78 (m, 9H), 1.30-1.40 (m, 1H).
Example 116
Preparation of
(E)-1-(3-(3-amino-3,3-dideutero-1-hydroxypropyl)styryl)cyclohexanol
##STR00417##
[1626]
(E)-1-(3-(3-Amino-3,3-dideutero-1-hydroxypropyl)styryl)cyclohexanol
was prepared following the method used in Example 114.
[1627] Step 1. Heck coupling between
N-(3-(3-bromophenyl)-1,1-dideuteropropyl)-2,2,2-trifluoroacetamide
and 1-vinylcyclohexanol following the method used in Example 114
gave
(E)-N-(1,1-dideutero-3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-
propyl)-2,2,2-trifluoroacetamide as a clear oil. Yield (0.41 g,
70%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.31 (s, 1H),
7.34 (s, 1H), 7.26-7.22 (m, 2H), 7.16-7.12 (m, 1H), 6.50 (d, J=16.0
Hz, 1H), 6.34 (d, J=16.0 Hz, 1H), 5.29 (d, J=4.4 Hz, 1H), 4.57-4.53
(m, 1H), 4.40 (s, 1H), 1.80-1.73 (m, 2H), 1.62-1.39 (m, 9H),
1.25-1.15 (m, 1H).
[1628] Step 2.
(E)-N-(1,1-dideutero-3-hydroxy-3-(3-(2-(1-hydroxycyclohexyl)vinyl)phenyl)-
propyl)-2,2,2-trifluoroacetamide was deprotected following the
method used in Example 114 to give Example 116 as a clear oil.
Yield (0.22 g, 72%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.36 (m, 1H), 7.22-7.15 (m, 2H), 7.14-7.11 (m, 1H), 6.50 (d, J=16.0
Hz, 1H), 6.33 (d, J=16.0 Hz, 1H), 4.63 (t, J=5.6 Hz, 1H), 4.40
(br.s, 1H), 1.67-1.52 (m, 4H), 1.49-1.39 (m, 7H), 1.26-1.17 (m,
1H); ESI MS m/z 278.2 [M+H].sup.+.
Example 117
Preparation of
(E)-4-(2-(3-(3-amino-1-hydroxypropyl)phenyl)-1,2-dideuterovinyl)heptan-4--
ol
##STR00418##
[1630]
(E)-4-(2-(3-(3-Amino-1-hydroxypropyl)phenyl)-1,2-dideuterovinyl)hep-
tan-4-ol was prepared following the method described below.
[1631] Step 1. To an ice cooled solution of
4-((3-(3-amino-1-hydroxypropyl)phenyl)ethynyl)heptan-4-ol (1.0 g,
3.46 mmol) in anhydrous ether was slowly added LiAlD.sub.4 (0.436
g, 10.4 mmol) over a 2-3 min period. The solution was allowed to
warm to room temp while stirring overnight. The reaction was
quenched with saturated solution of anhydrous Na.sub.2SO.sub.4 in
D.sub.2O (3 ml) and stirred for 6.0 hr. MgSO.sub.4 (.about.5 g) was
added and the solution was left to stand overnight. Filtration and
evaporation was followed with flash chromatography (10% 7N
NH.sub.3/MeOH/CH.sub.2Cl.sub.2) to give Example 117 as a clear oil.
Yield (0.524 g, 51%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.32 (m, 1H), 7.23-7.18 (m, 2H), 7.14-7.10 (m, 1H), 4.64-4.61 (m,
1H), 4.31 (br.s, 1H), 2.67-2.57 (m, 2H), 1.64-1.57 (m, 2H),
1.48-1.17 (m, 8H), 0.82 (t, J=7.2 Hz, 6H).
Example 118
Preparation of
(E)-1-(3-(3-amino-1-hydroxypropyl)-4-deuterostyryl)cyclohexanol
##STR00419##
[1633]
(E)-1-(3-(3-Amino-1-hydroxypropyl)-4-deuterostyryl)cyclohexanol was
prepared following the method shown in Scheme 14.
##STR00420##
[1634] Step 1: A mixture of 5-bromo-2-iodobenzaldehyde (14.9) (1.0
g, 3.2 mmol) and PTSA (0.1 g) in ethanol was stirred under reflux
for 18 hrs and concentrated under reduced pressure. The residue was
dissolved in ethyl acetate and washed with saturated NaHCO.sub.3,
dried over anhydrous Na.sub.2SO.sub.4 and concentrated to give
4-bromo-2-(diethoxymethyl)-1-iodobenzene (14.10) that was directly
used in next reaction without further purification.
[1635] Step 2. To a solution of
4-bromo-2-(diethoxymethyl)-1-iodobenzene (3.2 mmol) in THF was
added MeMgCl (2 ml, 3M in THF) at -25.degree. C. under argon. After
stirring at -25.degree. C. for 30 mins, the reaction mixture was
warmed to 0.degree. C. and stirred at 0.degree. C. for 30 mins.
D.sub.2O (0.6 ml) was added followed by 6N HCl (5 ml) and the
mixture was stirred at room temperature for 2 hrs, then extracted
with ethyl acetate (8 ml). Organic portion was washed with brine,
dried and concentrated to give product
3-bromo-5-deuterobenzaldehyde as a light yellow oil. Yield (0.59 g,
quant.); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.56 (s, 1H),
8.05 (d, J=2.0 Hz, 1H), 7.88 (dd, J=8.0, 2.4 Hz, 1H), 7.54 (d,
J=8.0 Hz, 1H).
[1636] Step 3: Addition of acetonitrile to
3-bromo-5-deuterobenzaldehyde (14.11) gave
3-(5-bromo-2-deuterophenyl)-3-hydroxypropanenitrile as a colorless
oil. Yield (0.31 g, 41%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.51 (d, J=2.0 Hz, 1H), 7.46 (dd, J=8.0, 2.0 Hz, 1H), 7.30
(d, J=8.0 Hz, 1H), 7.60 (t, J=1.6 Hz, 1H), 6.04 (br. s, 1H), 4.89
(br. s, 1H), 2.79-2.93 (m, 2H).
[1637] Step 4: A mixture of
3-(5-bromo-2-deuterophenyl)-3-hydroxypropanenitrile (14.12) (0.3 g,
1.32 mmol), borane-dimethylsulfide (0.5 mL, 3.9 mmol) in anhydrous
THF was sritted under reflux for 18 hr. After cooling to room
temperature MeOH was carefully added to the reaction mixture
followed by HCl/MeOH (1.25 M, 10 mL). The mixture was stirred at
50.degree. C. for 5 hrs and concentrated. To the residue was added
CH.sub.2Cl.sub.2-MeOH (2:1) (30 ml), CF.sub.3COOEt (5.0 mL) and
Et.sub.3N (2.0 mL, 14.3 mmol). The reaction mixture was stirred at
50.degree. C. for 8 h and concentrated under reduced pressure. The
residue was partitioned in EtOAc and 1N HCl. Organic portion was
washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and
concentrated under reduced pressure. Purification by flash
chromatography (40% to 50% EtOAc-hexanes gradient) gave
N-(3-(5-bromo-2-deuterophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamide
as a colorless oil. Yield (0.21 g, 89%); .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 9.16 (br.s, 1H), 7.53 (d, J=2.4 Hz, 1H), 7.39
(dd, J=8.0, 2.0 Hz, 1H), 7.23 (d, J=8.0 Hz, 1H), 4.65 (dd, J=7.6,
5.6 Hz, 1H), 3.35-3.41 (m, 2H), 1.88-1.94 (m, 2H).
[1638] Step 5. Heck coupling between
N-(3-(5-bromo-2-deuterophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamide
and 1-vinylcyclohexanol following the method used in Example 114
gave
(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(5-(2-(1-hydroxycyclohexyl)vinyl)-2-de-
uterophenyl)propyl)acetamide as a colorless oil. Yield (0.2 g,
84%); .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.39 (s, 1H),
7.26-7.28 (m, 2H), 6.60 (d, J=16.0 Hz, 1H), 6.36 (d, J=16.0 Hz,
1H), 4.67 (t, J=6.4 Hz, 1H), 3.37 (t, J=7.2 Hz, 2H), 1.94 (q, J=7.2
Hz, 2H), 1.49-1.76 (m, 9H), 1.26-1.40 (m, 1H).
[1639] Step 6.
(E)-2,2,2-trifluoro-N-(3-hydroxy-3-(5-(2-(1-hydroxycyclohexyl)vinyl)-2-de-
uterophenyl)propyl)acetamide (14) was deprotected following the
method used in Example 114 to give Example 118 as a light yellow
oil. Yield (0.15 g, quant.); .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.40 (s, 1H), 7.24-7.28 (m, 2H), 6.60 (d, J=16.4 Hz, 1H),
6.35 (d, J=16.4 Hz, 1H), 4.71 (dd, J=8.0, 5.6 Hz, 1H), 2.68-2.78
(m, 2H), 1.80-1.94 (m, 2H), 1.48-1.76 (m, 9H), 1.30-1.42 (m,
1H).
Example 119
Preparation of
4-((3-(3-amino-1-deutero-1-hydroxypropyl)phenyl)ethynyl)heptan-4-ol
##STR00421##
[1641]
4-((3-(3-Amino-1-deutero-1-hydroxypropyl)phenyl)ethynyl)heptan-4-ol
was prepared following the method shown in Scheme 15.
##STR00422##
[1642] Step 1: To a cold (-50.degree. C.) solution of
t-BuO.sup.-K.sup.+ in THF (1M, 0.76 L, 760 mmol) under N.sub.2 was
slowly added acetonitrile (37.0 mL, 703 mmol). The reaction mixture
was stirred for 25 min and then a solution of 3-bromobenzaldehyde
(15.1) (75 mL, 640 mmol) in anhydrous THF was added dropwise
keeping the temperature below -40.degree. C. After addition was
complete, the reaction mixture as stirred at for 45 min while
slowly warming to -10.degree. C. The reaction mixture was
partitioned between THF and an aqueous solution of NH.sub.4Cl
(25%), organic layer was washed with brine, dried over anhydrous
MgSO.sub.4 and filtered. The filtrate was concentrated under
reduced pressure to give hydroxynitrile 15.2 as an amber oil. Yield
(148 g, quant.); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.60
(t, J=1.6 Hz, 1H), 7.46 (ddd, J=7.6, 2.0, 1.2 Hz, 1H), 7.40 (dd,
J=7.6, 2.0 Hz, 1H), 7.31 (t, J=7.6 Hz, 1H), 6.05 (d, J=4.8 Hz, 1H),
4.87-4.92 (m, 1H), 2.94-2.80 (m, 2H).
[1643] Step 2: To an ice cold solution of
3-(3-bromophenyl)-3-hydroxypropanenitrile (15.2) (2.70 g, 11.9
mmol) in anhydrous THF under argon was added a solution of
LiAlH.sub.4 in THF (11.9 mL of a 2 M solution in THF, 23.8 mmol).
The mixture was stirred at 0.degree. C. for 45 min, diluted with
ether (50 mL), and quenched with the dropwise addition of saturated
aqueous Na.sub.2SO.sub.4 (approximately 2 mL). After drying over
MgSO.sub.4, the mixture was filtered and concentrated under reduced
pressure to give amine 15.3 as a light green oil. This material was
used in the next step without further purification. Yield (2.30 g,
84%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.49 (m, 1H),
7.37 (dt, J=7.2, 1.6 Hz, 1H), 7.23-7.31 (m, 2H), 4.66 (t, J=6.8 Hz,
1H), 2.61 (m, 2H), 1.61 (q, J=6.8 Hz, 2H).
[1644] Step 3: To a solution of amine 15.3 (5.67 g, 24.6 mmol) in
anhydrous CH.sub.2Cl.sub.2 was added Boc.sub.2O (5.69 g, 26.1
mmol). The reaction mixture was stirred at room temperature for 15
min, concentrated under reduced pressure, the residue was dissolved
in CH.sub.2Cl.sub.2 and Celite (8.67 g) followed by pyridinium
chlorochromate (7.67 g, 35.6 mmol) was added. The reaction mixture
was stirred at room temperature for 17 hrs and solvent was removed
under reduced pressure. Dark brown residue was suspended in
EtOAc-hexanes (30%), filtered and the filtrate was concentrated
under reduced pressure. Purification by flash chromatography (20%
to 80% EtOAc-hexanes gradient) gave ketone 15.4 as a light yellow
oil. Yield (7.2 g, 89%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.0-8.04 (m, 1H), 7.87-7.93 (m, 1H), 7.78-7.83)m, 1H), 7.47
(t, J=7.8 Hz, 1H), 6.78 (br. t, J=5.1 Hz, 1H), 3.25 (q, J=5.7 Hz,
2H), 3.12 (t, J=6.3 Hz, 2H), 1.33 (s, 9H).
[1645] Step 4: NaBD.sub.4 (1.07 g, 25.5 mmol) was added to stirred
solution of ketone 15.4 (3.30 g, 10.1 mmol) in i-PrOH. The reaction
mixture was stirred at room temperature for 30 min, aqueous
NH.sub.4Cl (25%) was carefully added. The product was extracted
with EtOAc, organic layer was washed with brine, dried over
anhydrous MgSO.sub.4 and concentrated under reduced pressure to
give 3-amino-1-(3-bromophenyl)-1-deuteropropan-1-ol as a colorless
oil. Yield (3.44 g, quant.); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.49 (t, J=1.6 Hz, 1H), 7.39 (dt, J=1.6, 7.4 Hz, 1H),
7.23-7.32 (m, 2H), 6.75 (br. t, J=4.9 Hz, 1H), 5.30 (s, 1H),
2.87-3.00 (m, 2H), 1.65 (t, J=7.0 Hz, 2H), 1.35 (s, 9H). A mixture
of 3-amino-1-(3-bromophenyl)-1-deuteropropan-1-ol (3.44 g),
HCl/i-PrOH (5.5 M, 30 mL) and Et.sub.2O was stirred at room
temperature for 6 hrs and concentrated under reduced pressure to
give amine hydrochloride 15.5 as a colorless oil. Yield (3.07 g,
quant.). The product was used in the next step without
purification.
[1646] Step 5: To a solution of salt 15.5 (3.07 g) in
CH.sub.2Cl.sub.2-MeOH (2:1) was added Et.sub.3N (1.8 mL, 12.9 mmol)
followed by CF.sub.3COOEt (3.0 mL, 25.1 mmol) and the reaction
mixture was stirred at room temperature overnight. The reaction
mixture was concentrated under reduced pressure, the residue was
partitioned between aq. NH.sub.4Cl (25%) and EtOAc. The aqueous
layer was extracted with EtOAc, combined organic layers were washed
with brine, dried over anhydrous MgSO.sub.4, and concentrated under
reduced pressure to give amide 15.6 as a light yellow oil. Yield
(3.14 g, 83%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.32
(br. s, 1H), 7.51 (t, J=1.8 Hz, 1H), 7.38-7.42 (m, 1H), 7.23-7.33
(m, 2H), 5.43 (s, 1H), 3.16-3.29 (m, 2H), 1.70-1.85 (m, 2H).
[1647] Step 6: A solution of alkyne 15.7 (0.657 g, 4.69 mmol) and
bromide 15.6 (1.369 g, 4.18 mmol) in Et.sub.3N (10 mL) was degassed
for 3 min by bubbling argon. CuI (0.04 g, 0.2 mmol) and
PdCl.sub.2(Ph.sub.3P).sub.2 (0.131 g, 0.19 mmol) were added, argon
was bubbled for 2 min and the reaction mixture was stirred under
argon at +80.degree. C. for 2 hrs. The reaction mixture was
concentrated under reduced pressure and the residue was purified by
flash chromatography (5% to 100% EtOAc-hexanes gradient). Fractions
containing product were pooled together, treated with activated
charcoal, filtered and the filtrate was concentrated under reduced
pressure to give alkyne 15.8 as a light yellow oil. Yield (1.35 g,
83.3%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.33 (br. s,
1H), 7.30-7.34 (m, 1H), 7.25-7.30 (m, 2H), 7.18-7.24 (m, 1H), 5.35
(s, 1H), 5.12 (s, 1H), 3.17-3.26 (m, 2H), 1.70-1.83 (m, 2H),
1.40-1.63 (m, 8H), 0.89 (t, J=7.0 Hz, 6H).
[1648] Step 7: A solution of amide 15.8 (0.619 g, 1.60 mmol) and
K.sub.2CO.sub.3 (0.909 g, 6.58 mmol) in MeOH:H.sub.2O (2:1, 18 mL)
was stirred at room temperature for 24 hrs and the reaction mixture
was concentrated under reduced pressure. Purification by flash
chromatography (20% to 100% of 20% 7N
NH.sub.3/MeOH-CH.sub.2Cl.sub.2--CH.sub.2Cl.sub.2 gradient) gave
Example 119 as a colorless oil. Yield (0.39 g, 84%); .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 7.39-7.41 (m, 1H), 7.26-7.33 (m, 3H),
2.68-2.79 (m, 2H), 1.76-1.89 (m, 2H), 1.52-1.73 (m, 8H), 0.97 (t,
J=7.0 Hz, 6H); RP-HPLC (Method 1) t.sub.R=9.16 min, 93.1% (AUC);
ESI-MS m/z 291.2 [M+H].sup.+.
Example 120
Preparation of
1-((3-(3-amino-2,2-dideutero-1-hydroxypropyl)phenyl)ethynyl)cyclohexanol
##STR00423##
[1650]
1-((3-(3-Amino-2,2-dideutero-1-hydroxypropyl)phenyl)ethynyl)cyclohe-
xanol was prepared following the method used in Example 119.
[1651] Step 1: Addition of trideuteroacetonitrile to
3-bromobenzaldehyde gave
3-(3-bromophenyl)-2,2-dideutero-3-hydroxypropanenitrile as a
colorless oil. Yield (5.17 g, 95%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.60 (t, J=1.6 Hz, 1H), 7.46 (ddd, J=1.2,
2.0, 7.8 Hz, 1H), 7.37-7.41 (m, 1H), 7.31 (t, J=7.6 Hz, 1H), 6.05
(d, J=4.0 Hz, 1H), 4.88 (m, 1H).
[1652] Step 2: A mixture of
3-(3-bromophenyl)-2,2-dideutero-3-hydroxypropanenitrile (1.91 g,
8.37 mmol), borane-dimethylsulfide (2.0 mL, 21.1 mmol) in anhydrous
THF was stirred under reflux for 15 hr. After cooling to room
temperature MeOH was carefully added to the reaction mixture
followed by HCl/MeOH (1.25 M, 10 mL). The mixture was stirred under
reflux for 4 hrs and concentrated under reduced pressure to give
3-amino-1-(3-bromophenyl)-2,2-dideuteropropan-1-ol hydrochloride as
a white foam which was used in the next step without purification.
Yield (2.25 g, quant.).
[1653] Step 3: To a solution of
3-amino-1-(3-bromophenyl)-2,2-dideuteropropan-1-ol hydrochloride
(2.25 g, 8.38 mmol) in CH.sub.2Cl.sub.2-MeOH (2:1) was added
CF.sub.3COOEt (3.0 mL) followed by Et.sub.3N (2.0 mL, 14.3 mmol).
The reaction mixture was stirred at room temperature for 1 h and
concentrated under reduced pressure. The residue was suspended in
EtOAc, washed with brine, dried over anhydrous MgSO.sub.4 and
concentrated under reduced pressure to give
N-(3-(3-bromophenyl)-2,2-dideutero-3-hydroxypropyl)-2,2,2-trifluoroacetam-
ide as a colorless oil. Yield (2.81 g, quant.); .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.32 (br.s, 1H), 7.49-7.52 (m, 1H), 7.40
(dt, J=1.6, 7.4 Hz, 1H), 7.24-7.32 (m, 2H), 5.44 (d, J=4.7 Hz, 1H),
4.56 (d, J=4.7 Hz, 1H), 3.16-3.27 (m, 2H).
[1654] Step 4: Sonogashira coupling between
N-(3-(3-bromophenyl)-2,2-dideutero-3-hydroxypropyl)-2,2,2-trifluoroacetam-
ide and 1-ethynylcyclohexanol following the method used in Example
119 gave
N-(2,2-dideutero-3-hydroxy-3-(3-((l-hydroxycyclohexyl)ethynyl)phenyl-
)propyl)-2,2,2-trifluoroacetamide as a light brown oil. Yield (0.99
g, 87%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.32 (br. t,
1H), 7.32-7.36 (m, 1H), 7.26-7.31 (m, 2H), 7.22-7.25 (m, 1H),
5.35-5.38 (m, 2H), 4.56 (d, J=4.5 Hz, 1H), 3.16-3.26 (m, 2H),
1.78-1.86 (m, 2H), 1.56-1.66 (m, 2H), 1.40-1.56 (m, 5H), 1.16-1.24
(m, 1H).
[1655] Step 5: Deprotection of
N-(2,2-dideutero-3-hydroxy-3-(3-((1-hydroxycyclohexyl)ethynyl)phenyl)prop-
yl)-2,2,2-trifluoroacetamide following the method used in Example
119 gave Example 120 as a colorless oil. Yield (0.22 g, 59%);
.sup.1H NMR ((400 MHz, DMSO-d.sub.6) .delta. 7.31-7.34 (m, 1H),
7.24-7.28 (m, 2H), 7.19-7.23 (m, 1H), 5.38 (br.s, 1H), 4.63 (s,
1H), 2.58 (dt, J=8.4, 12.0 Hz, 2H), 1.76-1.85 (m, 2H), 1.56-1.66
(m, 2H), 1.40-1.56 (m, 7H), 1.16-1.24 (m, 1H).
Example 121
Preparation of
1-((3-(3-amino-3,3-dideutero-1-hydroxypropyl)phenyl)ethynyl)cyclohexanol
##STR00424##
[1657]
1-((3-(3-Amino-3,3-dideutero-1-hydroxypropyl)phenyl)ethynyl)cyclohe-
xanol was prepared following the method used in Example 119.
[1658] Step 1: A solution of
3-(3-bromophenyl)-3-hydroxypropanenitrile (3.72 g, 16.5 mmol) in
anhydrous Et.sub.2O was added under argon to a cooled (0.degree.
C.) stirred suspension of LiAlD.sub.4 (0.76 g, 18.1 mmol) in
anhydrous Et.sub.2O and the reaction mixture was stirred at
0.degree. C. for 2 h. Saturated Na.sub.2SO.sub.4 was slowly added
to the reaction mixture until white precipitate formed. The
suspension was dried over anhydrous MgSO.sub.4 and filtered to give
a solution of 3-(3-bromophenyl)-1,1-dideuteropropan-1-amine.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.54 (t, J=1.6 Hz, 1H),
7.39 (dt, J=1.2, 7.8 Hz, 1H), 7.26-7.33 (m, 1H), 7.23 (t, J=7.8 Hz,
1H), 4.66 (dd, J=5.5, 7.4 Hz, 1H), 1.85-1.95 (m, 2H). Ethyl
trifluoroacetate (10 mL) was added to the solution of the amine and
the mixture was stirred at room temperature for 1 h, concentrated
under reduced pressure. Purification by flash chromatography (5% to
20% EtOAc-hexanes gradient) gave
N-(3-(3-bromophenyl)-1,1-dideutero-3-hydroxypropyl)-2,2,2-trifluoroacetam-
ide as a light yellow oil. Yield (3.76 g, 70%); .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 7.53 (br.t, J=1.6 Hz, 1H), 7.39 (ddd,
J=1.2, 1.8, 7.8 Hz, 1H), 1.30 (m, 1H), 7.23 (t, J=7.8 Hz, 1H), 4.66
(dd, J=5.5, 7.4 Hz, 1H), 1.85-1.94 (m, 2H).
[1659] Step 2: Sonogashira coupling between
N-(3-(3-bromophenyl)-1,1-dideutero-3-hydroxypropyl)-2,2,2-trifluoroacetam-
ide and 1-ethynylcyclohexanol following the method used in Example
120 gave
N-(1,1-dideutero-3-hydroxy-3-(3-((l-hydroxycyclohexyl)ethynyl)phenyl-
)propyl)-2,2,2-trifluoroacetamide as a light brown oil. Yield (0.84
g, 65%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.31 (br.s,
1H), 7.33-7.36 (m, 1H), 7.25-7.32 (m, 2H), 7.21-7.23 (m, 1H),
5.35-5.39 (m, 2H), 4.57 (dt, J=4.7, 7.8 Hz, 1H), 1.70-1.86 (m, 4H),
1.56-1.66 (m, 2H), 1.40-1.56 (m, 5H), 1.18-1.26 9m, 1H).
[1660] Step 3: Deprotection of
N-(1,1-dideutero-3-hydroxy-3-(3-((l-hydroxycyclohexyl)ethynyl)phenyl)prop-
yl)-2,2,2-trifluoroacetamide following the method used in Example
120 gave Example 121 as an off-white solid. Yield (0.097 g, 54%);
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.40-7.43 9m, 1H),
7.26-7.33 (m, 3H), 4.70 (dd, J=5.3, 7.8 Hz, 1H), 1.90-2.0 (m, 2H),
1.53-1.88 (m, 11H), 1.24-1.35 (m, 1H); RP-HPLC (Method 1)
t.sub.R=7.52 min, 96.7% (AUC); ESI-MS m/z 276.1 [M+H].sup.+.
Example 122
Preparation of
3-amino-1-(3-(cyclohexylethynyl)phenyl)-2,2-dideuteropropan-1-ol
##STR00425##
[1662]
3-Amino-1-(3-(cyclohexylethynyl)phenyl)-2,2-dideuteropropan-1-ol
was prepared following the method used in Examples 119, 120.
[1663] Step 1: Sonogashira coupling between
N-(3-(3-bromophenyl)-2,2-dideutero-3-hydroxypropyl)-2,2,2-trifluoroacetam-
ide and ethynylcyclohexane following the method used in Example 119
gave
N-(3-(3-(cyclohexylethynyl)phenyl)-2,2-dideutero-3-hydroxypropyl)-2,2,2-t-
rifluoroacetamide as a clear oil. Yield (0.29 g, 54%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.31 (s, 1H), 7.31 (s, 1H),
7.28-7.24 (m, 2H), 7.22-7.18 (m, 1H), 5.33 (d, J=4.4 Hz, 1H), 4.53
(d, J=4.0 Hz, 1H), 3.25-3.15 (m, 2H), 2.63-2.57 (m, 1H), 1.80-1.77
(m, 2H), 1.68-1.64 (m, 2H), 1.48-1.40 (m, 3H), 1.35-1.29 (m,
3H).
[1664] Step 2: Deprotection of
N-(3-(3-(cyclohexylethynyl)phenyl)-2,2-dideutero-3-hydroxypropyl)-2,2,2-t-
rifluoroacetamide following the method used in Example 119 gave
Example 122 as a colorless oil. Yield (0.14 g, 67%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.29 (s, 1H), 7.24-7.22 (m, 2H),
7.19-7.16 (m, 1H), 4.61 (s, 1H), 2.63-2.51 (m, 3H), 1.80-1.77 (m,
2H), 1.69-1.64 (m, 2H), 1.48-1.40 (m, 3H), 1.35-1.29 (m, 3H).
Example 123
Preparation of
3-amino-1-(3-(cyclohexylethynyl)phenyl)-3,3-dideuteropropan-1-ol
##STR00426##
[1666]
3-Amino-1-(3-(cyclohexylethynyl)phenyl)-3,3-dideuteropropan-1-ol
was prepared following the method used in Example 121, 119.
[1667] Step 1: Sonogashira coupling between
N-(3-(3-bromophenyl)-1,1-dideuteropropyl)-2,2,2-trifluoroacetamide
and ethynylcyclohexane following the method used in Example 119
gave
N-(3-(3-(cyclohexylethynyl)phenyl)-1,1-difluoro-3-hydroxypropyl)-2,2,2-tr-
ifluoroacetamide as a clear oil. Yield (0.079 g, 15%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.30 (s, 1H), 7.31 (s, 1H),
7.26-7.24 (m, 2H), 7.22-7.18 (m, 1H), 5.29 (d, J=4.0 Hz, 1H),
4.56-4.52 (m, 1H), 2.63-2.53 (m, 1H), 1.80-1.29 (m, 10H).
[1668] Step 2: Deprotection of
N-(3-(3-(cyclohexylethynyl)phenyl)-1,1-difluoro-3-hydroxypropyl)-2,2,2-tr-
ifluoroacetamide following the method used in Example 119 gave
Example 123 as a colorless oil. Yield (0.037 g, 73%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.29 (s, 1H), 7.24-7.22 (m, 2H),
7.20-7.16 (m, 1H), 4.62 (t, J=6.4 Hz, 1H), 2.63-2.56 (m, 1H),
1.80-1.77 (m, 2H), 1.69-1.62 (m, 2H), 1.57 (d, J=6.8 Hz, 2H),
1.52-1.40 (m, 3H), 1.35-1.29 (m, 3H).
Example 124
Preparation of
1-((3-(3-amino-1-hydroxypropyl)-4-deuterophenyl)ethynyl)cyclohexanol
##STR00427##
[1670]
1-((3-(3-Amino-1-hydroxypropyl)-4-deuterophenyl)ethynyl)cyclohexano-
l was prepared following the method shown in Scheme 16.
##STR00428##
[1671] Step 1: A mixture of 5-bromo-2-iodobenzaldehyde (1.0 g, 3.2
mmol) and PTSA (0.1 g) in ethanol was stirred under reflux for 18
hrs and concentrated under reduced pressure. The residue was
dissolved in ethyl acetate and washed with saturated NaHCO.sub.3,
dried over anhydrous Na.sub.2SO.sub.4 and concentrated to give
4-bromo-2-(diethoxymethyl)-1-iodobenzene (16.10) that was directly
used in next reaction without further purification.
[1672] Step 2. To a solution of
4-bromo-2-(diethoxymethyl)-1-iodobenzene (3.2 mmol) in THF was
added MeMgCl (2 ml, 3M in THF) at -25.degree. C. under argon. After
stirring at -25.degree. C. for 30 mins, the reaction mixture was
warmed to 0.degree. C. and stirred at 0.degree. C. for 30 mins.
D.sub.2O (0.6 ml) was added followed by 6N HCl (5 ml) and the
mixture was stirred at room temperature for 2 hrs, then extracted
with ethyl acetate (8 ml). Organic portion was washed with brine,
dried and concentrated to give product
3-bromo-5-deuterobenzaldehyde as a light yellow oil. Yield (0.59 g,
quant.); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.56 (s, 1H),
8.05 (d, J=2.0 Hz, 1H), 7.88 (dd, J=8.0, 2.4 Hz, 1H), 7.54 (d,
J=8.0 Hz, 1H).
[1673] Step 3: Addition of acetonitrile to
3-bromo-5-deuterobenzaldehyde (16.11) gave
3-(5-bromo-2-deuterophenyl)-3-hydroxypropanenitrile as a colorless
oil. Yield (0.31 g, 41%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.51 (d, J=2.0 Hz, 1H), 7.46 (dd, J=8.0, 2.0 Hz, 1H), 7.30
(d, J=8.0 Hz, 1H), 7.60 (t, J=1.6 Hz, 1H), 6.04 (br. s, 1H), 4.89
(br. s, 1H), 2.79-2.93 (m, 2H).
[1674] Step 4: A mixture of
3-(5-bromo-2-deuterophenyl)-3-hydroxypropanenitrile (16.12) (0.3 g,
1.32 mmol), borane-dimethylsulfide (0.5 mL, 3.9 mmol) in anhydrous
THF was stirred under reflux for 18 hr. After cooling to room
temperature MeOH was carefully added to the reaction mixture
followed by HCl/MeOH (1.25 M, 10 mL). The mixture was stirred at
50.degree. C. for 5 hrs and concentrated. To the residue was added
CH.sub.2Cl.sub.2-MeOH (2:1) (30 ml), CF.sub.3COOEt (5.0 mL) and
Et.sub.3N (2.0 mL, 14.3 mmol). The reaction mixture was stirred at
50.degree. C. for 8 h and concentrated under reduced pressure. The
residue was partitioned in EtOAc and 1N HCl. Organic portion was
washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and
concentrated under reduced pressure. Purification by flash
chromatography (40% to 50% EtOAc-hexanes gradient) gave
N-(3-(5-bromo-2-deuterophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamide
as a colorless oil. Yield (0.21 g, 89%); .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 9.16 (br.s, 1H), 7.53 (d, J=2.4 Hz, 1H), 7.39
(dd, J=8.0, 2.0 Hz, 1H), 7.23 (d, J=8.0 Hz, 1H), 4.65 (dd, J=7.6,
5.6 Hz, 1H), 3.35-3.41 (m, 2H), 1.88-1.94 (m, 2H).
[1675] Step 5. Sonogashira coupling between
N-(3-(5-bromo-2-deuterophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacetamide
and 1-ethynylcyclohexanol following the method used in Example 120
gave
2,2,2-trifluoro-N-(3-hydroxy-3-(5-((1-hydroxycyclohexyl)ethynyl)-2-deuter-
ophenyl)acetamide as a colorless oil. Yield (0.26 g, 88%); .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 7.41 (d, J=0.4 Hz, 1H), 7.28-7.30
(m, 2H), 4.66 (t, J=6.4 Hz, 1H), 3.37 (t, J=7.2 Hz, 2H), 1.90-1.98
(m, 4H), 1.54-1.78 (m, 7H), 1.24-1.34 (m, 1H).
[1676] Step 6.
2,2,2-Trifluoro-N-(3-hydroxy-3-(5-((1-hydroxycyclohexyl)ethynyl)-2-deuter-
ophenyl)acetamide (16.15) was deprotected following the method used
in Example 119 to give Example 124 as a light yellow oil. Yield
(0.15 g, 78%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.32 (d,
J=1.6 Hz, 1H), 7.27 (d, J=7.6 Hz, 1H), 7.19 (dd, J=7.6, 1.6 Hz,
1H), 5.37 (br s, 1H), 4.64 (t, J=6.4 Hz, 1H), 2.54-2.66 (m, 2H),
1.76-1.86 (m, 2H), 1.56-1.68 (m, 4H), 1.42-1.56 (m, 5H), 1.16-1.26
(m, 1H).
Example 125
Preparation of
1-((3-(3-amino-1-hydroxypropyl)-5-deuterophenyl)ethynyl)cyclohexanol
##STR00429##
[1678]
1-((3-(3-Amino-1-hydroxypropyl)-5-deuterophenyl)ethynyl)cyclohexano-
l is prepared following the method described below.
[1679] Step 1: A mixture of 3-bromo-5-iodophenol (1.40 g, 4.68
mmol), benzyl bromide (0.89 g, 5.20 mmol) and anhydrous
K.sub.2CO.sub.3 (1.44 g, 10.4 mmol) in anhydrous NMP (8 mL) was
stirred under argon at +70.degree. C. for 1 hour. The reaction
mixture was partitioned between aqueous NH.sub.4Cl and hexanes.
Aqueous layer was additionally extracted with hexanes and combined
organic layers were washed with 1N NaOH, brine, dried over
anhydrous MgSO.sub.4 and concentrated under reduced pressure to
give 1-(benzyloxy)-3-bromo-5-iodobenzene as a colorless oil. Yield
(2.14 g, 99%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.45 (t,
J=1.4 Hz, 1H), 7.32-7.40 (m, 5H), 7.26 (dd, J=1.4, 2.2 Hz, 1H),
7.09 (t, J=2.0 Hz, 1H), 5.00 (s, 2H).
[1680] Step 2: A solution of methylmagnesium chloride in THF (3N,
1.8 mL, 5.4 mmol) was added under argon to a cooled (-10.degree.
C.) solution of 1-(benzyloxy)-3-bromo-5-iodobenzene (1.82 g, 4.68
mmol) in anhydrous THF. The reaction mixture was stirred at
-10.degree. C. to 0.degree. C. for 2 hrs after which D.sub.2O (0.75
mL) was added to the reaction mixture. The mixture was stirred for
15 min and partitioned between NH.sub.4Cl and THF. Organic layer
was separated and concentrated under reduced pressure to give
1-(benzyloxy)-3-bromo-5-deuterobenzene as a light yellow oil. Yield
(1.29 g, quant.); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.30-7.45 (m, 5H), 7.14 (dd, J=1.8, 2.3 Hz, 1H), 7.07-7.10 (m, 1H),
6.88-6.91 (m, 1H), 5.04 (s, 2H).
[1681] Step 3: A solution of n-BuLi (2.5 M/THF, 3.0 mL, 7.5 mmol)
was added under argon to a cold (-78.degree. C.) solution of
1-(benzyloxy)-3-bromo-5-deuterobenzene (1.29 g, 4.88 mmol) and the
reaction mixture was stirred at -78.degree. C. for 10 min.
Anhydrous DMF (0.7 mL) was added to the reaction mixture and
stirring continued for 1 hr. Reaction was quenched by adding
aqueous NH.sub.4Cl. The mixture was stirred, layers were separated.
Aqueous layer was extracted with EtOAc. Combined organic layer were
washed with brine and concentrated under reduced pressure.
Purification by flash chromatography (1% to 20% EtOAc-hexanes
gradient) gave 3-(benzyloxy)-5-deuterobenzaldehyde as a white
solid. Yield (0.692 g, 67%); .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 9.97 (s, 1H), 7.28-7.48 (m, 8H), 5.12 (s, 2H).
[1682] Step 4: Addition of acetonitrile to
3-(benzyloxy)-5-deuterobenzaldehyde following the method used in
Example 6 gave
3-(3-(benzyloxy)-5-deuterophenyl)-3-hydroxypropanenitrile as a
yellow oil which was used in the next step without purification.
Yield (0.868 g, quant.); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.28-7.45 (m, 5H), 7.04-7.06 (m, 1H), 6.95-6.98 (m, 1H),
6.87-6.91 (m, 1H), 5.91 (d, J=4.5 Hz, 1H), 5.06 (s, 2H), 4.80-4.86
(m, 1H), 2.86 (ABd, J=4.9, 16.6 Hz, 1H), 2.77 (ABd, J=6.7, 16.6 Hz,
1H).
[1683] Step 5: Reduction of
3-(3-(benzyloxy)-5-deuterophenyl)-3-hydroxypropanenitrile following
the method used in Example 6 gave
3-amino-1-(3-(benzyloxy)-5-deuterophenyl)propan-1-ol hydrochloride
as a colorless oil which was used in the next step without further
purification. Yield (1.147 g, quant.).
[1684] Step 6: A mixture of
3-amino-1-(3-(benzyloxy)-5-deuterophenyl)propan-1-ol hydrochloride
(1.147 g, 3.89 mmol), Et.sub.3N (0.6 mL, 4.66 mmol), CF.sub.3COOEt
(0.7 mL, 5.87 mmol) in EtOH was stirred at room temperature for 1
hr. The reaction mixture was concentrated under reduced pressure
and the residue was resuspended in EtOAc. The resulting suspension
was filtered, the filtrate was concentrated under reduced pressure
to give crude
N-(3-(3-(benzyloxy)-5-deuterophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacet-
amide as a colorless oil which was used directly in the next step
without purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.32 (br.t, 1H), 7.27-7.44 (m, 5H), 6.95-6.97 (m, 1H), 6.86-6.88
(m, 1H), 6.83-6.85 (m, 1H), 5.30 (d, J=4.5 Hz, 1H), 5.06 (s, 2H),
4.49-4.55 (m, 1H), 3.18-3.25 (m, 2H), 1.72-1.81 (m, 2H).
[1685] Step 7: A solution of
N-(3-(3-(benzyloxy)-5-deuterophenyl)-3-hydroxypropyl)-2,2,2-trifluoroacet-
amide in EtOH was stirred under H.sub.2 atmosphere in the presence
of Pd(OH).sub.2/C (20% wt, 0.113 g) for 20 hrs. The reaction
mixture was filtered through Celite and concentrated under reduced
pressure. Purification by flash chromatography (20% to 100%
EtOAc-hexanes gradient) gave
2,2,2-trifluoro-N-(3-(3-deutero-5-hydroxyphenyl)-3-hydroxypropyl)ace-
tamide as a colorless oil. Yield (0.47 g, 46% over 2 steps);
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.32 (br. s, 1H), 9.24
(s, 1H), 6.66-6.74 (m, 2H), 6.56-6.60 (m, 1H), 5.22 (d, J=4.5 Hz,
1H), 4.42-4.50 (m, 1H), 3.17-3.25 (m, 2H), 1.68-1.80 (m, 2H).
[1686] Step 8: A mixture of
2,2,2-trifluoro-N-(3-(3-deutero-5-hydroxyphenyl)-3-hydroxypropyl)acetamid-
e, Et.sub.3N and triflic anhydride in anhydrous CH.sub.2Cl.sub.2 is
stirred at 0.degree. C. until no starting phenol is seen by TLC.
The reaction mixture is washed with brine, dried over anhydrous
MgSO.sub.4 and concentrated under reduced pressure. Purification by
flash chromatography (EtOAc-hexanes gradient) gives
3-deurero-5-(1-hydroxy-3-(2,2,2-trifluoroacetamido)propyl)phenyl
trifluoromethanesulfonate.
[1687] Step 9: Sonogashira coupling between
3-deurero-5-(1-hydroxy-3-(2,2,2-trifluoroacetamido)propyl)phenyl
trifluoromethanesulfonate and alkynol 14 following the method used
in Example 1 gives
2,2,2-trifluoro-N-(3-(3-deutero-5-((l-hydroxycyclohexyl)ethynyl)phenyl)-3-
-hydroxypropyl)acetamide.
[1688] Step 10: Deprotection of
2,2,2-trifluoro-N-(3-(3-deutero-5-((1-hydroxycyclohexyl)ethynyl)phenyl)-3-
-hydroxypropyl)acetamide following the method used in Example 1
gives Example 7.
Example 126
Preparation of
3-amino-1-(3-(cyclohexylmethoxy)phenyl)-1-deuteropropan-1-ol
##STR00430##
[1690] 3-Amino-1-(3-(cyclohexylmethoxy)phenyl)-1-deuteropropan-1-ol
was prepared following the method shown in Scheme 17.
##STR00431##
[1691] Step 1. To a mixture of 3-hydroxybenzaldehyde (545 g, 4.46
mol), K.sub.2CO.sub.3 (679 g, 4.91 mol) and NMP (0.718 L) was added
bromomethylcyclohexane (718 g, 4.05 mol) and the reaction mixture
was heated at +75.degree. C. for 24 hrs. The reaction mixture was
cooled to 20.degree. C. followed by addition of aqueous NaOH (1N),
water and heptane. The mixture was stirred for 15 min and layers
were separated. Organic layer was washed with NaOH (1N), brine and
concentrated under reduced pressure to give ether 17.3 as a pale
amber oil. Yield (675 g, 76%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.95 (s, 1H), 7.45-7.5 (m, 2H), 7.38-7.39 (m, 1H),
7.22-7.25 (m, 1H), 3.82 (d, J=6.4 Hz, 2H), 1.74-1.81 (m, 2H),
1.58-1.73 (m, 4H), 1.10-1.28 (m, 3H), 0.98-1.08 (m, 2H).
[1692] Step 2. Acetonitrile (118 mL, 2.26 mol) was added dropwise
under nitrogen to a cooled (-50.degree. C.) solution of potassium
tert-butoxide (1M/THF, 2.7 L, 2.7 mol). The reaction mixture was
stirred at -50.degree. C. for 40 mins and then a solution of
aldehyde 17.3 (450 g, 2.06 mol) in anhydrous THF was added dropwise
to the reaction mixture. The reaction mixture was stirred for 45
min at -45.degree. C. and cooling bath was replaced with ice bath.
The reaction mixture was stirred for 40 min after which aqueous
NH.sub.4Cl (20%) was added. Layers were separated and organic layer
was washed with brine, filtered and dried over anhydrous
Na.sub.2SO.sub.4. The mixture was concentrated under reduced
pressure to give hydroxynitrile 17.4 as amber oil. (Yield 502 g,
94%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.27-7.31 (m, 1H),
6.92-6.95 (m, 2H), 6.85-6.88 (m, 1H), 5.00 (t, J=6.4 Hz, 1H), 3.76
(d, J=6.4 Hz, 2H), 2.77 (d, J=1.6 Hz, 1H), 2.75 (s, 1H), 1.82-1.89
(m, 2H), 1.68-1.82 (m, 4H), 1.14-1.36 (m, 4H), 1.01-1.10 (m,
2H).
[1693] Step 3. Borane-dimethyl sulfide (240 mL, 2.52 mol) was added
dropwise under N.sub.2 atmosphere to a solution of nitrile (502 g,
3.55 mol) in anhydrous THF over 1 h while dimethylsulfide-THF (550
mL) was distilling off. The reaction mixture was heated under
reflux for 3 hrs, then cooled to 10.degree. C. and then aqueous HCl
(3N, 0.65 L) was slowly added. The mixture was stirred at room
temperature overnight, aqueous NaOH (50%) was added to pH 12. Water
and MTBE were added, the mixture was stirred and layers were
separated. Organic layer was washed with 30% NaCl, dried over
anhydrous Na.sub.2SO.sub.4 and concentrated under reduced pressure
Re-evaporation with absolute EtOH gave crude
3-amino-1-(3-(cyclohexylmethoxy)phenyl)propan-1-ol which was used
in the next step without further purification. Yield (504 g, 99%);
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.22 (t, J=8.0 Hz, 1H),
6.95 (t, J=1.6 Hz, 1H), 6.90 (d, J=7.6 Hz, 1H), 6.77 (ddd, J=8.0,
2.4, 0.8 Hz, 1H), 4.90 (dd, J=8.8, 3.2 Hz, 1H), 3.75 (d, J=6.4 Hz,
2H), 3.12 (br s, 2H), 3.06 (ddd, J=12.4, 6.0, 4.0 Hz, 1H),
2.90-2.96 (m, 1H), 1.82-1.89 (m, 3H), 1.67-1.81 (m, 6H), 1.15-1.34
(m, 3H), 0.99-1.09 (m, 2H).
[1694] To a solution of amine (504 g, 1.91 mol) in ethanol
ethanolic HCl (5.8 M, 266 mL) was added dropwise so that the
temperature was kept below +45.degree. C. The white precipitate
formed and the mixture was stirred at +40.degree. C. for 20 min.
The mixture was diluted with i-PrOAc and stirred for 20 min The
precipitate was collected by filtration, washed with i-PrOAc and
dried overnight under a stream of N.sub.2. Drying in vacuum gave
salt 5 as a white powder. Yield (425 g, 73%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.81 (br.s, 3H), 7.20 (t, J=7.8 Hz, 1H),
6.83-6.88 (m, 2H), 6.76 (ddd, J=0.8, 2.5, 8.2 Hz, 1H), 5.49 (d,
J=4.1 Hz, 1H), 4.58-4.66 (m, 1H), 3.73 (d, J=6.26 Hz, 2H),
2.74-2.86 (m, 2H), 1.59-1.90 (m, 8H), 0.95-1.30 (m, 5H).
[1695] Step 4: To a suspension of amine hydrochloride 17.5 (118 g,
0.396 mol) in anhydrous THF was added Et.sub.3N (42.0 g, 0.415 mol)
and Boc.sub.2O (86.3 g, 0.396 mol). The reaction mixture was
stirred overnight at room temperature, concentrated under reduced
pressure and partitioned between EtOAc and HCl (0.5 N). Organic
layer was washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure. Recrystallization of the
residue from hexanes/EtOAc gave carbamate 17.6 as a white solid.
Yield (125.4 g, 87%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.16 (t, J=7.8 Hz, 1H), 7.81-7.86 (m, 2H), 6.70-6.75 (m, 2H), 5.13
(d, J=4.5 Hz, 1H), 4.48 (q, J=4.9 Hz, 1H), 3.72 (d, J=6.26 Hz, 2H),
2.93 (q, J=6.8 Hz, 2H), 1.73-1.82 (m, 2H), 1.58-1.73 (m, 6H), 1.34
(s, 9H), 1.07-1.29 (m, 3H), 0.95-1.07 (m, 2H).
[1696] Step 5: To a solution of alcohol 17.6 (125.3 g, 345 mmol) in
dichloromethane was added Celite (125 g) and pyridinium
chlorochromate (81.8 g, 380 mmol). The mixture was stirred
overnight at room temp, filtered and the filtrate was concentrated
under reduced pressure. Purification by column chromatography (20%
EtOAc-hexanes) gave ketone 17.7 as a white solid. Yield (102 g,
82%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.45-7.50 (m,
1H), 7.35-7.42 (m, 2H), 7.14-7.18 (m, 1H), 6.77 (br.t, J=5.1 Hz,
1H), 3.80 (d, J=6.26 Hz, 2H), 3.24 (q, J=6.1 Hz, 2H), 3.10 (t,
J=6.5 Hz, 2H), 1.58-1.83 (m, 6H), 1.33 (s, 9H), 1.08-1.30 (m, 3H),
0.96-1.08 (m, 2H).
[1697] Step 6. Sodium borodeuteride (0.101 g, 2.41 mmol) was added
to a cooled (0.degree. C.) solution of ketone 7 (0.531 g, 1.47
mmol) in isopropanol and the reaction mixture was stirred at
0.degree. C. for 2 hrs. Aqueous NH.sub.4Cl (25%) was slowly added
to the reaction mixture followed by EtOAc. Layers were separated
and aqueous layer was additionally extracted with EtOAc. Combined
organic layers were washed with brine and dried over anhydrous
MgSO.sub.4. Concentration under reduced pressure gave alcohol 17.8
as a white solid. Yield (0.455 g, 85%).
[1698] Step 7. A solution of HCl in i-PrOH (5.5N, 3.0 mL) was added
to a stirred solution of carbamate 17.8 (0.454 g, 1.25 mmol) in
i-PrOAc at room temperature and the reaction mixture was stirred
for 20 hrs. The reaction mixture was concentrated under reduced
pressure, i-PrOAc was added to the residue and the mixture was
sonicated. The product was collected by filtration, washed with
i-PrOAc, hexanes and dried to give Example 126 hydrochloride as a
white solid. Yield (0.348 g, 93%); .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.16-7.26 (m, 1H), 6.85-6.94 (m, 2H), 6.74-6.82
(m, 1H), 3.73-3.78 (m, 2H), 2.98-3.14 (m, 2H), 1.93-2.07 (m, 2H),
1.66-1.90 (m, 5H), 1.16-1.40 (m, 3H), 1.02-1.16 (m, 2H); RP-HPLC
(Method 1) t.sub.R=10.05 min, 91.95% (AUC); ESI-MS m/z 265.2
[M+H].sup.+.
Example 127
Preparation of
3-amino-1-(3-(cyclohexylmethoxy)phenyl)-2,2-dideuteropropan-1-ol
##STR00432##
[1700]
3-Amino-1-(3-(cyclohexylmethoxy)phenyl)-2,2-dideuteropropan-1-ol
was prepared following the method shown in Scheme 18.
##STR00433##
[1701] Step 1. Addition of trideuteroacetonitrile to aldehyde 18.3
following the procedure shown in Example 126 gave hydroxynitrile
18.9 as a yellow oil. Yield (4.05 g, 85%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.21 (t, J=8.0 Hz, 1H), 6.90-6.96 (m, 2H),
6.80 (ddd, J=0.8, 2.4, 8.4 Hz, 1H), 5.88 (br. d, J=4.0 Hz, 1H),
4.81-4.82 (m, 1H), 3.74 (d, J=6.8 Hz, 2H), 1.58-1.83 (m, 6H),
1.09-1.29 (m, 3H), 0.95-1.07 (m, 2H).
[1702] Step 2. Reduction of hydroxynitrile 18.9 was done following
the procedure shown in Example 126 with the following exceptions.
Methanolic HCl (1.25 M, 3.68 mL, 4.6 mmol) was added to a cooled
solution (0.degree. C.) of free amine in Et.sub.2O. After stirring
for 15 min at 0.degree. C. the precipitate was collected by
filtration, washed with Et.sub.2O and dried to give Example 127
hydrochloride as a white solid. Yield (2.81 g, 61%); .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.95 (br.s, 3H), 7.20 (t, J=7.6 Hz,
1H), 6.83-6.88 (m, 2H), 6.76 (ddd, J=1.2, 2.4, 8.4 Hz, 1H), 5.49
(d, J=4.0 Hz, 1H), 4.62 (d, J=4.0 Hz, 1H), 3.73 (d, J=6.0 Hz, 2H),
2.54-2.57 (m, 2H), 1.58-1.82 (m, 6H), 1.08-1.28 (m, 3H), 0.95-1.07
(m, 2H); RP-HPLC (Method 1) t.sub.R=10.04 min, 91.95% (AUC); ESI-MS
m/z 266.2 [M+H].sup.+.
Example 128
Preparation of
3-amino-1-(3-(cyclohexylmethoxy)phenyl)-3,3-dideuteropropan-1-ol
##STR00434##
[1704]
3-Amino-1-(3-(cyclohexylmethoxy)phenyl)-3,3-dideuteropropan-1-ol
was prepared following the method shown in Scheme 19.
##STR00435##
[1705] Step 1. LiAlD.sub.4 was added under argon to a cooled
(0.degree. C.) solution of hydroxynitrile 19.4 (0.54 g, 2.08 mmol)
in anhydrous Et.sub.2O. The reaction mixture was stirred at
0.degree. C. for 40 min and quenched by slow addition of saturated
aqueous Na.sub.2SO.sub.4 until white precipitate formed. Anhydrous
MgSO.sub.4 was then added to the mixture which was stirred and
filtered. The filtrate was concentrated under reduced pressure, and
purification of the residue by flash column chromatography
(10%-100% of 20% 7N
NH.sub.3/MeOH/CH.sub.2Cl.sub.2--CH.sub.2Cl.sub.2 gradient) gave
pure amine as a colorless oil. Yield (0.346 g, 63%). The amine was
dissolved in i-PrOAc, cooled to 0.degree. C., and HCl/i-PrOH (5.5
N, 1 mL) was added to the reaction mixture. The precipitate was
collected by filtration, washed with i-PrOAc, hexanes and dried to
give Example 128 hydrochloride as a white solid. Yield (0.359 g,
91%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.23 (t, J=7.8
Hz, 1H), 6.89-6.94 (m, 2H), 6.79 (ddd, J=0.8, 2.4, 8.4 Hz, 1H),
4.79 (dd, J=4.4, 8.0 Hz, 1H), 3.76 (d, J=6.4 Hz, 2H), 1.90-2.04 (m,
2H), 1.82-1.90 (m, 2H), 1.66-1.80 (m, 4H), 1.16-1.38 (m, 3H),
1.02-1.14 (m, 2H); RP-HPLC (Method 1) t.sub.R=10.06 min, 97.5%
(AUC); ESI-MS m/z 266.2 [M+H].sup.+.
Example 129
Preparation of
3-amino-1-(3-((1-deuterocyclohexyl)methoxy)phenyl)propan-1-ol
##STR00436##
[1707]
3-Amino-1-(3-((1-deuterocyclohexyl)methoxy)phenyl)propan-1-ol was
prepared following the method shown in Scheme 20.
##STR00437##
[1708] Step 1. To a solution of 1-deuteroclohexanecarboxylic acid
(20.10) (5.0 g, 38.7 mmol) in anhydrous DMSO was added KOH (2.39 g,
42.6 mmol) with stirring for 5 min. Methyl iodide (6.59 g, 46.4
mmol) was added and the reaction mixture was stirred overnight at
room temperature. Saturated NaHCO.sub.3 and ether was added and the
mixture was washed with brine, dried over Na.sub.2SO.sub.4 and
evaporated to dryness giving methyl 1-deuterocyclohexanecarboxylate
(20.11) as a clear liquid. Yield (5.62 g, quant.); .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 3.55 (s, 3H), 1.78-1.75 (m, 2H),
1.65-1.60 (m, 2H), 1.57-1.52 (m, 1H), 1.34-1.09 (m, 5H).
[1709] Step 2. To a solution of ester 20.11 (5.0 g, 34.9 mmol) in
anhydrous CH.sub.2Cl.sub.2 on an ice bath was added a solution of
DIBAL-H in CH.sub.2Cl.sub.2 (1.0 M, 73.3 ml, 73.3 mmol) The
reaction mixture was allowed to warm to room temperature over 2 hrs
and quenched with Rochelle's salt (100 ml). The organic layer was
dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure
to give (1-deuterocyclohexyl)methanol (20.12) as a clear liquid.
Yield (3.99 g, 97%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
4.27 (t, J=5.2 Hz, 1H), 3.15 (d, J=5.2 Hz, 2H), 1.66-1.56 (m, 5H),
1.21-1.20 (m, 3H), 0.84-0.78 (m, 2H).
[1710] Step 3. To a solution of alcohol 20.12 (3.0 g, 26.0 mmol) in
anhydrous CH.sub.2Cl.sub.2 on an ice bath was added TEA (2.98 g,
28.6 mmol) and methanesulfonyl chloride (3.28 g, 28.6 mmol). The
reaction mixture was warmed to room temp over 2 hr. 1N HCl was
added and layers were separated. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to give
(1-deuterocyclohexyl)methyl methanesulfonate (20.13) as an off
white solid. Yield (4.92 g, 98%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 3.97 (s, 2H), 3.12 (s, 3H), 1.68-1.58 (m,
5H), 1.25-1.08 (m, 3H), 0.97-0.88 (m, 2H).
[1711] Step 4. Alkylation of 3-hydroxybenzaldehyde (20.2) by
mesylate 20.13 following the method shown in Example 126 gave
3-((1-deuterocyclohexyl)methoxy)benzaldehyde (20.14) as a colorless
oil. Yield (0.47 g, 55%); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.94 (s, 1H), 7.50-7.44 (m, 2H), 7.39-7.38 (m, 1H), 7.24
(dt, J=6.8, 2.4 Hz, 1H), 3.82 (s, 2H), 1.79-1.61 (m, 5H), 1.23-0.91
(m, 5H).
[1712] Step 5. Acetonitrile addition to aldehyde following the
method shown in Example 126 gave
3-(3-((l-deuterocyclohexyl)methoxy)phenyl)-3-hydroxypropanenitrile
(20.15) as a colorless oil. Yield (0.53 g, 96%); .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 7.21 (t, J=7.8 Hz, 1H), 6.94-6.91 (m,
2H), 6.80 (ddd, J=8.4, 2.4, 0.8 Hz, 1H), 5.88, (d, J=4.4 Hz, 1H),
4.84-4.80 (m, 1H), 3.73 (s, 2H), 2.85 (Abd, J=16.8, 4.8 Hz, 1H),
2.77 (Abd, J=16.4, 5.2 Hz, 1H), 1.79-1.61 (m, 5H), 1.28-0.94 (m,
5H).
[1713] Step 6. Hydroxynitrile reduction following the method shown
in Example 126 gave free amine as a colorless oil. Amine was
converted into HCl salt following the method shown in Example 126
to give Example 129 hydrochloride as a white solid. Yield (0.27 g,
44%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.91 (br.s, 3H),
7.20 (t, J=7.8 Hz, 1H), 6.86-6.84 (m, 2H), 6.76 (m, 1H), 5.50, (d,
J=4.4 Hz, 1H), 4.65-4.60 (m, 1H), 3.72 (s, 2H), 2.78-2.80 (m, 2H),
1.89-1.61 (m, 7H), 1.27-0.94 (m, 5H); RP-HPLC (Method 1)
t.sub.R=10.04 min, 96.9% (AUC); ESI-MS m/z 265.2 [M+H].sup.+.
Example 130
Preparation of
(R)-3-amino-1-(3-(cyclohexyldideuteromethoxy)phenyl)propan-1-ol
##STR00438##
[1715]
(R)-3-Amino-1-(3-(cyclohexyldideuteromethoxy)phenyl)propan-1-ol was
prepared following the method shown in Schemes 21a and 21b.
##STR00439##
[1716] Step 1: To a stirred suspension of t-BuO.sup.-K.sup.+ (68.5
g, 614 mmol) in THF, cooled to -50.degree. C., was added
acetonitrile (30.3 mL, 540 mmol), dropwise over a period of 5 min.
The resulting mixture was stirred at -50.degree. C. for 30 min
following which a solution of 3-hydroxybenzaldehyde (21.2) (30.0 g,
244 mmol) in THF was added slowly, over a period of 10 min. This
was then allowed to warm to 0.degree. C. and stirred for another 3
h during which the reaction was complete. The reaction was quenched
by slow addition of ice-water followed by extraction with EtOAc.
The combined organics were washed with water, brine and dried over
Na.sub.2SO.sub.4. The solution was concentrated under reduced
pressure to give 3-hydroxy-3-(3-hydroxyphenyl)propanenitrile
(21.16) as yellow oil which was purified by flash column
chromatography (0 to 20% EtOAc-hexanes gradient). Yield (25.0 g,
62%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.27 (s, 1H), 6.95
(d, J=7.6 Hz, 1H), 6.90-6.93 (m, 1H), 6.82 (dd, J=8.0, 2.4 Hz, 1H),
4.91-5.03 (m, 1H), 2.76 (d, J=6.4 Hz, 2H).
[1717] Step 2: To a stirred solution of the nitrile 21.16 (25.0 g,
153 mmol) in THF, cooled to 0.degree. C., was added BH.sub.3-DMS
(49.5 mL, 460 mmol), following which the cooling bath was removed.
The resulting mixture was boiled under reflux overnight, cooled in
an ice-bath and quenched by the slow addition of large excess of
MeOH. After stirring at room temperature for 2 h, the excess
solvent was removed under reduced pressure. The residue was again
treated with MeOH and evaporated. The process was repeated three
times. The brown oil was then applied onto a flash silica gel
column and eluted (0 to 15% (9:1 MeOH-NH.sub.3)-DCM gradient) to
give 3-(3-amino-1-hydroxypropyl)phenol (21.17) as a brown solid.
Yield (25.0 g, 97%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.04-7.09 (m, 1H), 6.74 (s, 1H), 6.70 (d, J=7.6 Hz, 1H), 6.58 (dd,
J=8.0, 2.0 Hz, 1H), 4.55 (dd, J=7.2, 5.6 Hz, 1H), 2.57-2.66 (m,
2H), 1.56-1.62 (m, 2H).
[1718] Step 3: To a solution of amine 21.17 (25.0 g, 0.149 mol) in
1,4-dioxane was added K.sub.2CO.sub.3 (20.6 g, 150 mmol) followed
by the slow addition of Boc.sub.2O (36 mL, 150 mmol). The mixture
was stirred at room temperature for 2 h during which the reaction
was found to be complete. This mixture was then quenched by the
addition of water and extracted with ethyl acetate. The organic
layer was washed with water and brine. This was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. Purification by flash chromatography (0 to 20%
EtOAc-hexanes gradient) afforded crude tert-butyl
3-hydroxy-3-(3-hydroxyphenyl)propylcarbamate (21.18) as off white
solid. Yield (35.0 g, quant); 1H NMR (400 MHz, CDCl.sub.3) .delta.
7.05-7.10 (m, 1H), 6.70-6.76 (m, 2H), 6.59 (dd, J=8.0, 1.6 Hz, 1H),
5.11 (d, J=4.4 Hz, 1H), 4.42-4.47 (m, 1H), 3.57 (s, 1H), 2.92-2.98
(m, 2H), 1.61-1.67 (m, 2H), 1.37 (s, 9H).
[1719] Step 4: A stirred suspension of PCC (42.3 g, 196 mmol) and
Celite (43 g) in DCM (300 mL) was cooled to 0.degree. C. Alcohol
21.18 (35.0 g, 131 mmol) was slowly added to the reaction mixture
over a period of 15 min. The reaction mixture was allowed to stir
at room temperature for 2 h. The reaction mixture was then filtered
through a pad of Celite and the filter bed was washed with DCM.
Concentration of the filtrate gave a black tarry mass which was
purified by flash chromatography (30-50% ethyl acetate-hexanes
gradient) to give tert-butyl
3-(3-hydroxyphenyl)-3-oxopropylcarbamate 21.19 as pale yellow
solid. Yield (20.3 g, 58%); .sup.1H NMR (400 MHz, CDCl3) .delta.
9.78 (s, 1H), 7.27-7.40 (m, 2H), 7.01 (dd, J=8.0, 1.6 Hz, 1H),
6.80-6.83 (m, 1H), 3.22-3.27 (m, 2H), 3.08 (t, J=6.8 Hz, 2H), 1.36
(s, 9H).
[1720] Step 5: To a stirred solution of TFA (80 mL) and DCM at
0.degree. C. was slowly added ketone (20 g, 75 mmol). The resulting
reaction mixture was allowed to stir at room temperature for 2 h.
After the reaction was complete, the solvent was removed under
reduced pressure and the residue was triturated with toluene. The
complete removal of the solvent gave the TFA salt of amine. The
crude mass was directly utilized for the next transformation
without purification. Yield (21.0 g, crude); MS 166
[M+H].sup.+.
[1721] DIPEA (23 mL, 179 mmol) was added to a cooled to 0.degree.
C. solution of crude amine (21.0 g, 72 mmol) in a mixture of
acetonitrile:toluene (1:3). The resulting mixture was stirred at
room temperature for 10 min. This was followed by the addition of
phthalic anhydride (10.6 g, 72 mmol). The reaction mixture was then
refluxed for 2 h using a Dean-Stark assembly. After completion of
the reaction the solvent was distilled off under reduced pressure
and the reaction mass was treated with DCM. The organic layer was
washed with water and saturated NH.sub.4Cl, followed by saturated
NaHCO.sub.3, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give phthalimidophenol 21.20
as an off-white solid. Yield (14 g, 62%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 9.79 (s, 1H), 7.82-7.88 (m, 4H), 7.38 (d, J=8.0
Hz, 1H), 7.31 (d, J=7.6 Hz, 1H), 7.28 (s, 1H), 7.01 (dd, J=8.0, 2.0
Hz, 1H), 3.91 (t, J=7.2 Hz, 2H), 3.37 (t, J=7.2 Hz, 2H). MS: 296
[M+1].sup.+.
[1722] Step 6: A solution of (+)-diisopinocampheylchloroborane
((+)-Ipc.sub.2B-Cl) in hexanes (1.5 M, 14 mL, 21 mmol) was added
under inert atmosphere to a solution of ketone 21.20 (3.02 g, 10.2
mmol) in anhydrous THF at room temperature. The reaction mixture
was stirred for 3.5 hrs and partitioned between 25% NH.sub.4Cl and
THF. Aqueous layer was additionally extracted with EtOAc, combined
organic layers were washed with brine, dried over anhydrous
MgSO.sub.4, and concentrated under reduced pressure. Purification
by flash chromatography (15% to 60% EtOAc-hexanes gradient) gave
(R)-alcohol 21.21 as a white solid. Yield (2.78 g, 92%); .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 9.23 (s, 1H), 7.75-7.84 (m,
4H), 7.04 (t, J=7.6 Hz, 1H), 6.67-6.73 (m, 2H), 6.54 (ddd, J=1.0,
2.3, 8.0 Hz, 1H), 5.22 (d, J=4.3 Hz, 1H), 4.49 (dt, J=4.5, 6.3 Hz,
1H), 3.55-3.69 (m, 2H), 1.85 (q, J=7.4 Hz, 2H).
##STR00440##
[1723] Step 7. A solution of ester 21.22 (9.99 g, 70.3 mmol) was
added under inert atmosphere to a cooled (0.degree. C.) suspension
of LiAlD.sub.4 (2.99 g, 71.2 mmol) in anhydrous Et.sub.2O. The
reaction mixture was stirred at 0.degree. C. for 3 hrs and then
slowly quenched by addition of saturated Na.sub.2SO.sub.4 until
white precipitate formed. The mixture was dried over anhydrous
MgSO.sub.4, filtered. The filtrate was concentrated under reduced
pressure to give alcohol 21.23 as a colorless volatile liquid.
Yield (2.52 g, 32%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
1.63-1.78 (m, 5H), 1.40-1.50 (m, 1H), 1.10-1.35 (m, 4H), 0.86-0.99
(m, 2H).
[1724] Step 8. Mesylation of alcohol 21.23 following the method
used in Example 129 gave mesylate 22.24 as a colorless oil. Yield
(4.14 g, 97%); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 2.98 (s,
3H), 1.64-1.80 (m, 6H), 1.10-1.32 (m, 3H), 0.92-1.05 (m, 2H).
[1725] Step 9. NaH (60% suspension in mineral oil, 0.98 g, 2.45
mmol) was added to a stirred solution of phenol 21.21 (0.756 g,
2.54 mmol) in anhydrous DMSO. The mixture was stirred at room
temperature until allNaH dissolved. Mesylate 21.24 was added to the
resulting yellow solution of phenolate and the reaction mixture was
stirred at +90.degree. C. under argon for 2 days. The reaction
mixture was partitioned between EtOAc and 25% NH.sub.4Cl, aqueous
layer was extracted with EtOAc. Combined organic layers were washed
with brine, dried over anhydrous MgSO.sub.4 and concentrated under
reduced pressure. Purification by flash chromatography (5% to 50%
EtOAc-hexanes gradient) gave ether 21.25 as a colorless oil. Yield
(0.25 g, 27%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.81 (m,
4H), 7.15 (t, J=8.0 Hz, 1H), 6.80-6.90 (m, 2H), 6.65-6.73 (m, 1H),
5.25-5.29 (m, 1H), 4.52-4.60 (m, 1H), 3.56-3.73 (m, 2H), 1.84-1.94
(m, 2H), 1.57-1.84 (m, 6H), 1.10-1.30 (m, 3H), 0.96-1.08 (m,
2H).
[1726] Step 10. A mixture of phthalimide 21.25 (0.24 g, 0.607
mmol), N.sub.2H.sub.4.H.sub.2O (0.15 mL) in EtOH was stirred at
room temperature for 26 hrs. The reaction mixture was concentrated
under reduced pressure; the residue was resuspended in
CH.sub.2Cl.sub.2, filtered. The filtrate was dissolved in i-PrOAc
(20 mL), cooled to 0.degree. C. and HCl/i-PrOH (5.5M, 0.4 mL) was
added. The precipitate was collected by filtration to give Example
130 hydrochloride as a white solid. Yield (0.126 g, 69%); .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 7.23 (t, J=7.8 Hz, 1H), 6.88-6.95
(m, 2H), 6.77-6.82 (m, 1H), 4.79 (dd, J=4.5, 7.6 Hz, 1H), 2.97-3.11
(m, 2H), 1.91-2.03 (m, 2H), 1.81-1.90 (m, 2H), 1.66-1.80 (m, 4H),
1.161-1.37 (m, 3H), 1.02-1.14 (m, 2H); RP-HPLC (Method 1)
t.sub.R=9.96 min, 90.7% (AUC); ESI-MS m/z 266.2 [M+H].sup.+.
Example 131
Preparation of
3-amino-1-(3-((perdeuterocyclohexyl)methoxy)phenyl)propan-1-ol
##STR00441##
[1728]
3-Amino-1-(3-((perdeuterocyclohexyl)methoxy)phenyl)propan-1-ol was
prepared following the method used in Example 129.
[1729] Step 1. Reaction between perdeuterocyclohexylcarboxylic acid
and MeI gave methyl perdeuterocyclohexanecarboxylate as a clear
liquid. Yield (2.26 g, quant.); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 3.55 (s).
[1730] Step 2. Reduction of methyl perdeuterocyclohexanecarboxylate
with DIBAL-H gave (perdeuterocyclohexyl)methanol as a clear oil.
Yield (1.86 g, quant.); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
4.26 (t, J=5.2 Hz, 1H), 3.15 (d, J=5.2 Hz, 2H).
[1731] Step 3. Mesylation of (perdeuterocyclohexyl)methanol gave
(perdeuterocyclohexyl)methyl methanesulfonate as a pale yellow
liquid. Yield (3.02 g, quant.); .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 3.97 (s, 2H), 3.12 (s, 3H).
[1732] Step 4. 3-((Perdeuterocyclohexyl)methoxy)benzaldehyde was
prepared following the method used in Example 4. Yield (1.32 g,
40%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.94 (s, 1H),
7.50-7.44 (m, 2H), 7.39-7.38 (m, 1H), 7.23 (dt, J=2.4, 6.8 Hz, 1H),
3.81 (s, 2H).
[1733] Step 5.
3-(3-((Perdeuterocyclohexyl)methoxy)phenyl)-3-hydroxypropanenitrile
was prepared following the method used in Example 129. Yield (1.47
g, 96%); .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.21 (t, J=7.8
Hz, 1H), 6.94-6.91 (m, 2H), 6.80 (ddd, J=8.4, 2.4, 0.8 Hz, 1H),
5.88, (d, J=4.4 Hz, 1H), 4.84-4.80 (m, 1H), 3.73 (s, 2H), 2.85
(ABd, J=16.8, 4.8 Hz, 1H), 2.77 (ABd, J=16.4, 5.2 Hz, 1H).
[1734] Step 6. Hydroxynitrile reduction following the method used
in Example 129 gave, after column chromatography purification (10%
MeOH/CH.sub.2Cl.sub.2 followed by 10% 7N
NH.sub.3/MeOH/CH.sub.2Cl.sub.2)
3-amino-1-(3-((perdeuterocyclohexyl)methoxy)phenyl)propan-1-ol as a
colorless oil. Yield (1.06 g, 71%). The amine was dissolved in
Et.sub.2O, cooled on ice bath and HCl/MeOH (1.25M, 3.7 mL, 4.6
mmol) was added. The mixture was stirred for 15 min, the
precipitate was collected by filtration to give Example 131
hydrochloride as a white solid. Yield (0.72 g, 61%); m.p.
165-166.degree. C.; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.20 (t, J=7.8 Hz, 1H), 6.87-6.84 (m, 2H), 6.76 (m, 1H), 5.88, (d,
J=4.4 Hz, 1H), 4.64-4.61 (m, 1H), 3.72 (s, 2H), 2.85-2.74 (m, 2H),
1.91-1.76 (m, 2H); RP-HPLC (Method 2) t.sub.R=4.29 min, 99.4%
(AUC); ESI-MS m/z 275.3 [M+H].sup.+; Elemental analysis: C, 61.7%;
H, 8.32%; N, 4.57%; Cl, 11.42%.
Example 132
Preparation of
3-amino-1-(3-(cyclohexylmethoxy)-5-deuterophenyl)propan-1-ol
##STR00442##
[1736] 3-Amino-1-(3-(cyclohexylmethoxy)-5-deuterophenyl)propan-1-ol
was prepared following the method shown in Scheme 23.
##STR00443##
[1737] Step 1. Alkylation of 3-bromo-5-iodophenol (23.26) with
bromomethylcyclohexane following the method used in Example 126
gave ether 23.27 as a colorless oil. Yield (2.30 g, 87%); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.40 (t, J=1.6 Hz, 1H), 7.16 (dd,
J=1.4, 2.2 Hz, 1H), 6.99 (dd, J=1.8, 2.2 Hz, 1H), 3.68 (d, J=6.26
Hz, 2H), 1.66-1.86 (m, 6H), 1.16-1.37 (m, 3H), 0.96-1.10 (m,
2H).
[1738] Step 2. To a cold (-25.degree. C.) solution of iodide 23.27
(1.95 g, 4.94 mmol) under argon was added a solution of MeMgCl in
THF (3N, 2.0 mL, 6.0 mmol) and the reaction mixture was slowly
warmed to 0.degree. C. D.sub.2O (0.6 mL) was added to the reaction
mixture which was stirred for additional 20 min while warming to
room temperature. The mixture was partitioned between aqueous
NH.sub.4Cl (25%) and THF. Aqueous layer was extracted with EtOAc,
combined organic layers were washed with brine, dried over
anhydrous MgSO.sub.4 and concentrated under reduced pressure to
give deuteride 23.28 as a colorless oil. Yield (1.56 g, quant.);
.sup.1H NMR (400 MHz, CDCl3) .delta. 7.11 (t, J=8.2 Hz, 1H),
7.02-7.06 9m, 2H), 3.72 (d, J=6.3 Hz, 2H), 1.65-1.88 (m, 6H),
1.12-1.35 (m, 3H), 0.97-1.09 (m, 2H).
[1739] Step 3. To a cold (-78.degree. C.) solution of
1-bromo-3-(cyclohexylmethoxy)-5-deuterobenzene (23.28) (1.56 g,
5.77 mmol) under argon in anhydrous THF (10 mL) was added a
solution of n-BuLi in hexanes (2.5 M, 3.0 mL, 7.5 mmol) and the
reaction mixture was stirred at -78.degree. C. for 20 min. DMF (1.0
mL, 23 mmol) was added, the reaction mixture was allowed to warm to
-20.degree. C. and partitioned between aqueos NH.sub.4Cl (25%, mL)
and EtOAc. Aqueous layer was extracted with EtOAc, combined organic
layers were washed with brine, dried over anhydrous MgSO.sub.4, and
concentrated under reduced pressure. The residue was purified to
give 3-(cyclohexylmethoxy)-5-deuterobenzaldehyde (23.29) as a
colorless oil. Yield (0.97 g, 77%); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 10.01 (s, 1H), 7.41-7.44 (m, 1H), 7.37 (dd,
J=1.4, 2.7 Hz, 1H), 7.15-7.17 (m, 1H), 3.80 (d, J=6.3 Hz, 2H),
1.66-1.90 (m, 6H), 1.14-1.36 (m, 3H), 1.00-1.11 (m, 2H).
[1740] Step 4. Acetonitrile addition to aldehyde 23.29 following
the method used in Example 126 gave hydroxypropanenitrile 23.30 as
a colorless oil. Yield (1.09 g, 95%); .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.90-6.96 (m, 2H), 6.77-6.81 (m, 1H), 5.88
(d, J=4.5 Hz, 1H), 4.80-4.85 (m, 1H), 3.74 (d, J=6.3 Hz, 2H), 2.86
(ABd, J=4.9, 16.8 Hz, 1H), 2.77 (ABd, J=6.8, 16.8 Hz, 1H),
1.60-1.82 (m, 6H), 1.10-1.30 (m, 3H), 0.95-1.08 (m, 2H).
[1741] Step 5.
3-(3-(Cyclohexylmethoxy)-5-deuterophenyl)-3-hydroxypropanenitrile
(23.30) was reduced with borane following the method used in
Example 126 except the following. After the reduction was complete
as judged by TLC (50% EtOAc-hexanes), MeOH was slowly added to the
reaction mixture until a gas formation ceased, followed by HCl/MeOH
(1.25 M, 8 mL). The mixture was heated under reflux for 1.5 hrs and
concentrated under reduced pressure. The residue was crystallized
from i-PrOH/EtOAc (1:2) to give Example 132 hydrochloride as a
white solid. Yield (0.96 g, 79%); .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 6.90-6.94 (m, 2H), 6.78-6.81 (m, 1H), 4.79 (dd, J=4.7, 7.4
Hz, 1H), 3.76 (d, J=6.3 Hz, 2H), 2.96-3.11 (m, 2H), 1.90-2.04 (m,
2H), 1.82-1.90 (m, 2H), 1.66-1.81 (m, 4H), 1.16-1.38 (m, 3H),
1.02-1.13 (m, 2H); RP-HPLC (Method 1) t.sub.R=10.07 min, 97.8%
(AUC); ESI-MS m/z 265.2 [M+H].sup.+.
Example 133
In Vitro Isomerase Inhibition Assay
[1742] The capability of compounds described herein to inhibit the
activity of a visual cycle isomerase was determined in vitro either
in a human or bovine-based assay system. The isomerase inhibition
reactions were performed essentially as described (Stecher et al.,
J. Biol. Chem. 274:8577-85 (1999); see also Golczak et al., Proc.
Natl. Acad. Sci. USA 102:8162-67 (2005), reference 3), either using
a human cell line or a bovine retinal pigment epithelium (RPE)
microsome membranes as the source of visual enzymes.
Isolation of Human Apo Cellular Retinaldehyde-Binding Protein
(CRALBP)
[1743] Recombinant human apo cellular retinaldehyde-binding protein
(CRALBP) was cloned and expressed according to standard methods in
the molecular biology art (see Crabb et al., Protein Science
7:746-57 (1998); Crabb et al., J. Biol. Chem. 263:18688-92 (1988)).
Briefly, total RNA was prepared from confluent ARPE19 cells
(American Type Culture Collection, Manassas, Va.), cDNA was
synthesized using an oligo(dT).sub.12-18 primer, and then DNA
encoding CRALBP was amplified by two sequential polymerase chain
reactions (see Crabb et al., J. Biol. Chem. 263:18688-92 (1988);
Intres, et al., J. Biol. Chem. 269:25411-18 (1994); GenBank
Accession No. L34219.1). The PCR product was sub-cloned into
pTrcHis2-TOPO TA vector according to the manufacturer's protocol
(Invitrogen Inc., Carlsbad, Calif.; catalog no. K4400-01), and then
the sequence was confirmed according to standard nucleotide
sequencing techniques. Recombinant 6.times.His-tagged human CRALBP
was expressed in One Shot TOP 10 chemically competent E. coli cells
(Invitrogen), and the recombinant polypeptide was isolated from E.
coli cell lysates by nickel affinity chromatography using nickel
(Ni) Sepharose XK16-20 columns for HPLC (Amersham Bioscience,
Pittsburgh, Pa.; catalog no. 17-5268-02). The purified
6.times.His-tagged human CRALBP was dialyzed against 10 mM
bis-tris-Propane (BTP) and analyzed by SDS-PAGE. The molecular
weight of the recombinant human CRALBP was approximately 39
kDal.
Human In Vitro Isomerase Inhibition Reaction
[1744] The concentration dependent effect of the compounds
disclosed herein on the retinol isomerization reaction was
evaluated with a recombinant human enzyme system. In particular,
the in vitro isomerase assay was performed essentially as in
Golczak et al. 2005 (Proc. Natl. Acad. Sci. USA 102:8162-67 (2005),
reference 3). A homogenate of HEK293 cell clone expressing
recombinant human RPE65 and LRAT were the source of the visual
enzymes, and exogenous all-trans-retinol (about 20 .mu.M) was used
as the substrate. Recombinant human CRALBP (about 80 ug/mL) was
added to enhance the formation of 11-cis-retinal. The 200 .mu.L
Bis-Tris Phosphate buffer (10 mM, pH 7.2) based reaction mixture
also contains 0.5% BSA, and 1 mM NaPPi. In this assay, the reaction
was carried out at 37.degree. C. in duplicates for one hour and was
terminated by addition of 300 .mu.L methanol. The amount of
reaction product, 11-cis-retinol, was measured by HPLC analysis
following Heptane extraction of the reaction mixture. The Peak Area
Units (PAUs) corresponding to 11-cis-retinol in the HPLC
chromatograms were recorded and concentration dependent curves
analyzed by GraphPad Prism for IC.sub.50 values. The ability of the
compounds disclosed herein to inhibit isomerization reaction was
quantified and the respective IC.sub.50 value was determined. Table
2 summarizes the IC.sub.50 values of several of the compounds of
the present disclosure. FIGS. 1 and 2 depict dose-dependent curves
for the inhibition of the accumulation of 11-cis-retinol in the
human in vitro assay by the compounds of Example 5 and Example 6
(Compound 5 and Compound 6).
TABLE-US-00005 TABLE 2 Human in vitro Inhibition Data IC.sub.50
(nM) Compound/Example Number >1 to .ltoreq.10 nM 35, 37, 81, 91,
117, 120, 121, 122, 123, 126, 127, 128, 129, 130, 131, 132 >10
to .ltoreq.100 nM 5, 11, 12, 13, 14, 15, 33, 39, 40, 41, 47, 48,
49, 51, 58, 82, 83, 90, 93, 114, 115, 116, 118, 119, 124 >100 to
.ltoreq.1000 nM 4, 6, 7, 8, 16, 17, 18, 20, 21, 22, 31, 52, 57, 60
>1000 nM 1, 2, 3, 9, 10, 64, 99
Bovine In Vitro Isomerase Inhibition Reaction
[1745] Bovine RPE microsome membrane extracts are prepared
according to methods described (Golczak et al., Proc. Natl. Acad.
Sci. USA 102:8162-67 (2005)) and stored at about -80.degree. C.
Crude RPE microsome extracts are thawed in a 37.degree. C. water
bath, and then immediately placed on ice. About 50 ml crude RPE
microsomes are placed into a 50 ml Teflon-glass homogenizer (Fisher
Scientific, catalog no. 0841416M) on ice, powered by a hand-held
DeWalt drill, and homogenized about ten times up and down on ice
under maximum speed. This process is repeated until the crude RPE
microsome solution is homogenized. The homogenate is then subjected
to centrifugation (50.2 Ti rotor (Beckman, Fullerton, Calif.),
about 13,000 RPM; about 15360 Rcf) for about 15 minutes at
4.degree. C. The supernatant is collected and subjected to
centrifugation at about 42,000 RPM (about 160,000 Rcf; 50.2 Ti
rotor) for about 1 hour at 4.degree. C. The supernatant is removed,
and the pellets are suspended in about 12 ml (final volume) cold 10
mM MOPS buffer, pH 7.0. The resuspended RPE membranes in about 5 ml
aliquots are homogenized in a glass-to-glass homogenizer (Fisher
Scientific, catalog no.K885500-0021) to high homogeneity. Protein
concentration is quantified using the BCA protein assay according
to the manufacturer's protocol (Pierce, Rockford, Ill.). The
homogenized RPE preparations are stored at -80.degree. C.
[1746] Compounds described herein and control compounds are
reconstituted in ethanol to about 0.1 M. Ten-fold serial dilutions
(10.sup.-1, 10.sup.-2, 10.sup.-3, 10.sup.-4, 10.sup.-5,
10.sup.-6'10.sup.-7 M) in ethanol of each compound are prepared for
analysis in the isomerase assay.
[1747] The isomerase assay is performed in about 10 mM
bis-tris-propane (BTP) buffer, pH .about.7.5, .about.0.5% BSA
(diluted in BTP buffer), about 1 mM sodium pyrophosphate, about 20
.mu.M all-trans-retinol (in ethanol), and about 6 .mu.M apo-CRALBP.
The test compounds (.about.2 .mu.l) (final 1/15 dilution of serial
dilution stocks) are added to the above reaction mixture to which
RPE microsomes are added. The same volume of ethanol is added to
the control reaction (absence of test compound). Bovine RPE
microsomes (.about.9 .mu.l) (see above) are then added, and the
mixtures transferred to 37.degree. C. to initiate the reaction
(total volume=.about.150 .mu.l). The reactions are stopped after
about 30 minutes by adding methanol (about 300 .mu.l). Heptane is
added (300 .mu.l) and mixed into the reaction mixture by pipetting.
Retinoid is extracted by agitating the reaction mixtures, followed
by centrifugation in a microcentrifuge. The upper organic phase is
transferred to HPLC vials and then analyzed by HPLC using an
Agilent 1100 HPLC system with normal phase column: SILICA (Agilent
Technologies, dp 5 g, 4.6 mmX, 25CM; running method has a flow rate
of 1.5 ml/min; injection volume about 100 .mu.l). The solvent
components are about 20% of about 2% isopropanol in EtOAc and about
80% of 100% hexane.
[1748] The area under the A.sub.318 nm curve represents the
11-cis-retinol peak, which is calculated by Agilent Chemstation
software and recorded manually. The IC.sub.50 values (concentration
of compound that gives 50% inhibition of 11-cis-retinol formation
in vitro) are calculated using GraphPad Prism.RTM. 4 Software
(Irvine, Calif.). All tests are performed in at least duplicate and
it is expected that the compounds of the present disclosure show
concentration dependent effects on the retinol isomerization
reaction, as compared to control compounds.
Example 134
In Vivo Murine Isomerase Assay
[1749] The capability of compounds described herein to inhibit
isomerase was determined by an in vivo murine isomerase assay.
Brief exposure of the eye to intense light ("photobleaching" of the
visual pigment or simply "bleaching") is known to photo-isomerize
almost all 11-cis-retinal in the retina. The recovery of
11-cis-retinal after bleaching can be used to estimate the activity
of isomerase in vivo. Delayed recovery, as represented by lower
11-cis-retinal oxime levels, indicates inhibition of isomerization
reaction. Procedures were performed essentially as described by
Golczak et al., Proc. Natl. Acad. Sci. USA 102:8162-67 (2005). See
also Deigner et al., Science, 244: 968-71 (1989); Gollapalli et
al., Biochim Biophys Acta. 1651: 93-101 (2003); Parish, et al.,
Proc. Natl. Acad. Sci. USA, 14609-13 (1998); Radu, et al., Proc
Natl Acad Sci USA 101: 5928-33 (2004).
[1750] About six-week old dark-adapted CD-1 (albino) male mice were
orally gavaged with compound (0.01-25 mg/kg) dissolved in an
appropriate amount of oil (about 100 .mu.l corn oil containing 10%
ethanol, at least five animals per group). Mice were gavaged with
the compounds described in the present disclosure. After about 2-24
hours in the dark, the mice were exposed to photobleaching of about
5,000 lux of white light for 10 minutes. The mice were allowed to
recover for about 2 hours in the dark. The animals were then
sacrificed by carbon dioxide inhalation. Retinoids were extracted
from the eye and the regeneration of 11-cis-retinal was assessed at
various time intervals.
Eye Retinoid Extraction
[1751] All steps were performed in darkness with minimal redlight
illumination (low light darkroom lights and red filtered
flashlights for spot illumination as needed) (see, e.g., Maeda et
al., J. Neurochem 85:944-956, 2003; Van Hooser et al., J Biol Chem
277:19173-82, 2002). After the mice were sacrificed, the eyes were
immediately removed and placed in liquid nitrogen for storage.
[1752] The eyes were placed in about 500 .mu.L of bis-tris propane
buffer (10 mM, pH .about.7.3) and about 20 .mu.L of 0.8M
hydroxylamine (pH-7.3). The eyes were cut up into small pieces with
small iris scissors and then thoroughly homogenized at 30000 rpm
with a mechanical homogenizer (Polytron PT 1300 D) in the tube
until no visible tissue remains. About 500 .mu.L of methanol and
about 500 .mu.L of heptane was added to each tube. The tubes were
attached to a vortexer so that the contents are mixed thoroughly
for about 15 minutes in room temperature. The organic phase was
separated from the aqueous phase by centrifugation for about 10 min
at 13K rpm, 4.degree. C. 240 .mu.L of the solution from the top
layer (organic phase) was removed and transferred to clean 300
.mu.l glass inserts in HPLC vials using glass pipette and the vials
were crimped shut tightly.
[1753] The samples were analyzed on an Agilent 1100 HPLC system
with normal phase column: SILICA (Beckman Coutlier, dp 5 .mu.m, 4.6
mM.times.250 mM). The running method has a flow rate of 1.5 ml/min;
solvent components are 15% solvent 1 (1% isopropanol in ethyl
acetate), and 85% solvent 2 (100% hexanes). Loading volume for each
sample was about 100 .mu.l; detection wavelength is 360 nm. The
area under the curve for 11-cis-retinal oxime was calculated by
Agilent Chemstation software and recorded manually. Data processing
was performed using Prizm software.
[1754] Positive control mice (no compound administered) were
sacrificed fully dark-adapted and the eye retinoids analyzed. Light
(bleached) control mice (no compound administered) were sacrificed
and retinoids isolated and analyzed immediately after light
treatment.
[1755] A time course study was also performed to determine the
isomerase inhibitory activity of compounds of the present
disclosure. Female or male mice (such as Balb/c mice) (at least
4/group) received 0 to about 5 mg of compounds (in water) per kg
bodyweight orally, by gavage. The animals were then
"photo-bleached" (about 5000 Lux white light for about 10 minutes)
at about 2, 4, 8, 16 and 24 hours after dosing, and returned to
darkness to allow recovery of the 11-cis-retinal content of the
eyes. Mice were sacrificed about 2 hours after bleaching, eyes were
enucleated, and retinoid content was analyzed by HPLC.
[1756] A dose response in vivo isomerase inhibition study is
performed with compounds of the present disclosure. Male or female
mice (such as Balb/c mice)(at least about 8/group) are dosed orally
with about 0.01 to 25 mg/kg of the compounds of HCl salts of the
compounds in sterile water as solution, and photobleached about 4
hours after dosing. Recovery and retinoid analysis is performed as
described above. Dark control mice are vehicle-only treated,
sacrificed fully dark adapted without light treatment, and
analyzed. The concentration-dependent inhibition of isomerase
activity at about 4 hours post dosing of the compounds, inhibition
of 11-cis-retinal (oxime) recovery for and estimates of ED.sub.50s
(dose of compound that gives 50% inhibition of 11-cis-retinal
(oxime) recovery) are calculated. Table 3 provides the in vivo
inhibition data.
TABLE-US-00006 TABLE 3 In vivo Inhibition Data Example % Inibition
% Inibition Number 1 mg/kg, 24 h 1 mg/kg, 4 h 5 Not tested -11.7
.+-. 4.36 6 48.70 .+-. 2.71 -11.93 .+-. 18.17 15 Not tested -0.003
.+-. 19.4 11 Not tested 95.27 .+-. 2.7 13 Not tested 1.979 .+-.
6.016 131 Not tested 97.9 .+-. 11.8 126 Not tested 91.75 .+-. 2.7
128 Not tested 98.0 .+-. 0.99 121 Not tested 27.5 .+-. 9.6 129 Not
tested 97.23 .+-. 1.5 130 Not tested 100.9 .+-. 0.955 132 Not
tested 100.8 .+-. 1.2 117 Not tested 97.9 .+-. 1.5 123 Not tested
91.4 .+-. 2.5 122 Not tested 84.9 .+-. 4.5 35 Not tested 95.25 .+-.
1.41 33 Not tested 4.32 .+-. 7.88 40 Not tested 1.24 .+-. 9.74 39
Not tested 69.94 .+-. 6.85 57 Not tested 2.01 .+-. 1.3 31 Not
tested 9.52 .+-. 4.6 47 Not tested 4.08 .+-. 4.84 58 Not tested
6.94 .+-. 5.15 16 Not tested 17.08 .+-. 5.32 14 Not tested 8.12
.+-. 16.18 12 Not tested 9.16 .+-. 9.41 93 Not tested -0.53 .+-.
4.53 83 Not tested 89.46 .+-. 2.09 81 Not tested 84.98 .+-. 3.06 90
Not tested 3.17 .+-. 4.97 91 Not tested 95.49 .+-. 1.07 82 Not
tested 75.08 .+-. 8.03 60 Not tested -1.33 .+-. 5.20 48 Not tested
0.21 .+-. 8.88 41 Not tested -0.34 .+-. 6.12 99 Not tested -3.83
.+-. 5.52 37 Not tested 101.56 .+-. 0.49 64 Not tested 3.83 .+-.
3.83 49 Not tested 4.54 .+-. 6.31 73 Not tested -1.24 .+-. 4.43 59
Not tested 4.24 .+-. 12.99 36 Not tested -1.4 .+-. 2.78 103 Not
tested -2.77 .+-. 6.53 101 Not tested 1.82 .+-. 10.54
[1757] A single dose study of any compound is also performed at
various dosages, a various time points post dosing. The experiments
can be carried out in CD1 male mice, by way of example. Results are
analyzed by HPLC. It is expected that the compounds of the present
disclosure will exhibit different profiles of activity at different
times and dosages, with different compounds also exhibiting
different recovery patterns.
Example 135
Preparation of Retinal Neuronal Cell Culture System
[1758] This example describes methods for preparing a long-term
culture of retinal neuronal cells. All compounds and reagents can
be obtained from Sigma Aldrich Chemical Corporation (St. Louis,
Mo.) or other suitable vendors.
Retinal Neuronal Cell Culture
[1759] Porcine eyes are obtained from Kapowsin Meats, Inc. (Graham,
Wash.). Eyes are enucleated, and muscle and tissue are cleaned away
from the orbit. Eyes are cut in half along their equator and the
neural retina is dissected from the anterior part of the eye in
buffered saline solution, according to standard methods known in
the art. Briefly, the retina, ciliary body, and vitreous are
dissected away from the anterior half of the eye in one piece, and
the retina is gently detached from the clear vitreous. Each retina
is dissociated with papain (Worthington Biochemical Corporation,
Lakewood, N.J.), followed by inactivation with fetal bovine serum
(FBS) and addition of 134 Kunitz units/ml of DNaseI. The
enzymatically dissociated cells are triturated and collected by
centrifugation, resuspended in Dulbecco's modified Eagle's medium
(DMEM)/F12 medium (Gibco BRL, Invitrogen Life Technologies,
Carlsbad, Calif.) containing about 25 .mu.g/ml of insulin, about
100 .mu.g/ml of transferrin, about 60 .mu.M putrescine, about 30 nM
selenium, about 20 nM progesterone, about 100 U/ml of penicillin,
about 100 .mu.g/ml of streptomycin, about 0.05 M Hepes, and about
10% FBS. Dissociated primary retinal cells are plated onto
Poly-D-lysine- and Matrigel-(BD, Franklin Lakes, N.J.) coated glass
coverslips that are placed in 24-well tissue culture plates (Falcon
Tissue Culture Plates, Fisher Scientific, Pittsburgh, Pa.). Cells
are maintained in culture for 5 days to one month in 0.5 ml of
media (as above, except with only 1% FBS) at 37.degree. C. and 5%
CO.sub.2.
Immunocytochemistry Analysis
[1760] The retinal neuronal cells are cultured for about 1, 3, 6,
and 8 weeks, and the cells are analyzed by immunohistochemistry at
each time point. Immunocytochemistry analysis is performed
according to standard techniques known in the art. Rod
photoreceptors are identified by labeling with a rhodopsin-specific
antibody (mouse monoclonal, diluted about 1:500; Chemicon,
Temecula, Calif.). An antibody to mid-weight neurofilament (NFM
rabbit polyclonal, diluted about 1:10,000, Chemicon) is used to
identify ganglion cells; an antibody to .beta.3-tubulin (G7121
mouse monoclonal, diluted about 1:1000, Promega, Madison, Wis.) is
used to generally identify interneurons and ganglion cells, and
antibodies to calbindin (AB1778 rabbit polyclonal, diluted about
1:250, Chemicon) and calretinin (AB5054 rabbit polyclonal, diluted
about 1:5000, Chemicon) are used to identify subpopulations of
calbindin- and calretinin-expressing interneurons in the inner
nuclear layer. Briefly, the retinal cell cultures are fixed with 4%
paraformaldehyde (Polysciences, Inc, Warrington, Pa.) and/or
ethanol, rinsed in Dulbecco's phosphate buffered saline (DPBS), and
incubated with primary antibody for about 1 hour at 37.degree. C.
The cells are then rinsed with DPBS, incubated with a secondary
antibody (Alexa 488- or Alexa 568-conjugated secondary antibodies
(Molecular Probes, Eugene, Oreg.)), and rinsed with DPBS. Nuclei
are stained with 4', 6-diamidino-2-phenylindole (DAPI, Molecular
Probes), and the cultures are rinsed with DPBS before removing the
glass coverslips and mounting them with Fluoromount-G (Southern
Biotech, Birmingham, Ala.) on glass slides for viewing and
analysis.
[1761] Survival of mature retinal neurons after varying times in
culture is indicated by the histochemical analyses. Photoreceptor
cells are identified using a rhodopsin antibody; ganglion cells are
identified using an NFM antibody; and amacrine and horizontal cells
are identified by staining with an antibody specific for
calretinin.
[1762] Cultures are analyzed by counting rhodopsin-labeled
photoreceptors and NFM-labeled ganglion cells using an Olympus IX81
or CZX41 microscope (Olympus, Tokyo, Japan). Twenty fields of view
are counted per coverslip with a 20.times. objective lens. Six
coverslips are analyzed by this method for each condition in each
experiment. Cells that are not exposed to any stressor are counted,
and cells exposed to a stressor are normalized to the number of
cells in the control. It is expected that compounds presented in
this disclosure promote dose-dependent and time-dependent survival
of mature retinal neurons.
Example 136
Effect of Compounds on Retinal Cell Survival
[1763] This Example describes the use of the mature retinal cell
culture system that comprises a cell stressor for determining the
effects of a compound on the viability of the retinal cells.
[1764] Retinal cell cultures are prepared as described in Example
135. A2E is added as a retinal cell stressor. After culturing the
cells for 1 week, a chemical stress, A2E, is applied. A2E is
diluted in ethanol and added to the retinal cell cultures at
concentration of about 0, 10 .mu.M, 20 .mu.M, and 40 .mu.M.
Cultures are treated for about 24 and 48 hours. A2E is obtained
from Dr. Koji Nakanishi (Columbia University, New York City, N.Y.)
or is synthesized according to the method of Parish et al. (Proc.
Natl. Acad. Sci. USA 95:14602-13 (1998)). A compound described
herein is then added to the culture. To other retinal cell
cultures, a compound described herein is added before application
of the stressor or is added at the same time that A2E is added to
the retinal cell culture. The cultures are maintained in tissue
culture incubators for the duration of the stress at 37.degree. C.
and 5% CO.sub.2. The cells are then analyzed by immunocytochemistry
as described in Example 135.
Apoptosis Analysis
[1765] Retinal cell cultures are prepared as described in Example
135 and cultured for about 2 weeks and then exposed to white light
stress at about 6000 lux for about 24 hours followed by about a
13-hour rest period. A device was built to uniformly deliver light
of specified wavelengths to specified wells of the 24-well plates.
The device contains a fluorescent cool white bulb (GE P/N
FC12T9/CW) wired to an AC power supply. The bulb is mounted inside
a standard tissue culture incubator. White light stress is applied
by placing plates of cells directly underneath the fluorescent
bulb. The CO.sub.2 levels are maintained at about 5%, and the
temperature at the cell plate is maintained at 37.degree. C. The
temperature is monitored by using thin thermocouples. The light
intensities for all devices is measured and adjusted using a light
meter from Extech Instruments Corporation (P/N 401025; Waltham,
Mass.). A compound described herein is added to wells of the
culture plates prior to exposure of the cells to white light and is
added to other wells of the cultures after exposure to white light.
To assess apoptosis, TUNEL is performed as described herein.
[1766] Apoptosis analysis is also performed after exposing retinal
cells to blue light. Retinal cell cultures are cultured as
described in Example 135. After culturing the cells for about 1
week, a blue light stress is applied. Blue light is delivered by a
custom-built light-source, which consists of two arrays of 24
(4.times.6) blue light-emitting diodes (Sunbrite LED P/N
SSP-01TWB7UWB12), designed such that each LED is registered to a
single well of a 24 well disposable plate. The first array is
placed on top of a 24 well plate full of cells, while the second
one is placed underneath the plate of cells, allowing both arrays
to provide a light stress to the plate of cells simultaneously. The
entire apparatus is placed inside a standard tissue culture
incubator. The CO.sub.2 levels are maintained at about 5%, and the
temperature at the cell plate is maintained at about 37.degree. C.
The temperature is monitored with thin thermocouples. Current to
each LED is controlled individually by a separate potentiometer,
allowing a uniform light output for all LEDs. Cell plates are
exposed to about 2000 lux of blue light stress for about either 2
hours or 48 hours, followed by about a 14-hour rest period. A
compound described herein is added to wells of the culture plates
prior to exposure of the cells to blue light and is added to other
wells of the cultures after exposure to blue light. To assess
apoptosis, TUNEL is performed as described herein.
[1767] To assess apoptosis, TUNEL is performed according to
standard techniques practiced in the art and according to the
manufacturer's instructions. Briefly, the retinal cell cultures are
first fixed with 4% paraformaldehyde and then ethanol, and then
rinsed in DPBS. The fixed cells are incubated with TdT enzyme (0.2
units/.mu.l final concentration) in reaction buffer (Fermentas,
Hanover, Md.) combined with Chroma-Tide Alexa568-5-dUTP (0.1 .mu.M
final concentration) (Molecular Probes) for about 1 hour at
37.degree. C. Cultures are rinsed with DPBS and incubated with
primary antibody either overnight at 4.degree. C. or for about 1
hour at 37.degree. C. The cells are then rinsed with DPBS,
incubated with Alexa 488-conjugated secondary antibodies, and
rinsed with DPBS. Nuclei are stained with DAPI, and the cultures
are rinsed with DPBS before removing the glass coverslips and
mounting them with Fluoromount-G on glass slides for viewing and
analysis.
[1768] Cultures are analyzed by counting TUNEL-labeled nuclei using
an Olympus IX81 or CZX41 microscope (Olympus, Tokyo, Japan). Twenty
fields of view are counted per coverslip with a 20.times. objective
lens. Six coverslips are analyzed by this method for each
condition. Cells that are not exposed to a compound described
herein are counted, and cells exposed to the antibody are
normalized to the number of cells in the control. Data are analyzed
using the unpaired Student's t-test. It is expected that compounds
described herein reduce A2E-induced apoptosis and cell death in
retinal cell cultures in a dose-dependent and time-dependent
manner.
[1769] The cells are assessed for cell death using Sytox green
nucleic acid stain assay (Sytox, Molecular Probes, Eugene, Oreg.).
Sytox is a DNA-binding dye that penetrates only dying cells in
which the plasma membrane is compromised. The green nucleic acid
stain assay is added at 1 .mu.M to 96-well plates and incubated for
30 minutes at 37.degree. C. Fluorescence is determined using a
plate reader with excitation fluorescence at 485 nm and emission
fluorescence at 528 nm.
Example 137
In Vivo Light Mouse Model
[1770] This Example describes the effect of a compound in an in
vivo light damage mouse model.
[1771] Exposure of the eye to intense white light can cause
photo-damage to the retina. The extent of damage after light
treatment can be evaluated by measuring cytoplasmic
histone-associated-DNA-fragment (mono- and oligonucleosomes)
content in the eye (see, e.g., Wenzel et al., Prog. Retin. Eye Res.
24:275-306 (2005)).
[1772] Dark adapted mice (for example, male Balb/c (albino,
10/group)) are gavaged with the compounds of the present disclosure
at various doses (about 0.01-25 mg/kg) or vehicle only is
administered. About six hours after dosing, the animals are
subjected to light treatment (8,000 lux of white light for 1 hour).
Mice are sacrificed after about 40 hours of recovery in dark, and
retinas are dissected. A cell death detection ELISA assay is
performed according to the manufacturer's instructions (ROCHE
APPLIED SCIENCE, Cell Death Detection ELISA plus Kit). Contents of
fragmented DNA in the retinas are measured to estimate the
retinal-protective activity of the compounds. It is expected that
compounds of the present disclosure mitigate or inhibit
photo-damage to the retina.
Example 138
Electroretinographic (ERG) Study
[1773] This example describes determining the effect of a compound
that is a visual cycle modulator on the magnitude of the ERG
response in the eyes of mice after oral dosing of the animals with
the compound. The level of ERG response in the eyes is determined
after administering the compound to the animals (for example at 18
and 66 hours post administration).
[1774] Three groups of about nine-week old mice (19-25 grams), both
genders (strain C5 7BL/6, Charles River Laboratories, Wilmington,
Mass.) are housed at room temperature, 72.+-.4.degree. F., and
relative humidity of approximately 25%. Animals are housed in a
12-hour light/dark cycle environment, have free access to feed and
drinking water and are checked for general health and well-being
prior to use and during the study. Body weights are determined for
a representative sample of mice prior to initiation of dosing. The
average weight determined from this sampling is used to establish
the dose for all mice in the study.
[1775] Each test compound is dissolved in the control solvent
(EtOH), and diluted 1:10 (90 ml/900 ml) in the appropriate oil (for
example corn oil (Crisco Pure Corn Oil, J. M. Smucker Company,
Orrville, Ohio)) to the desired dose (mg/kg) in the desired volume
(about 0.1 mL/animal). The control vehicle is ethanol: oil (about
1:10 (0.9 ml/9 ml)). An example of treatment designations and
animal assignments are described in Table 4.
TABLE-US-00007 TABLE 4 Group Route Treatment Dose (mg/kg) Animals
Test oral test compound (~0.01-~25 mg/kg) >4 Control oral
Vehicle None >4
[1776] Animals are dosed once orally by gavage, with the assigned
vehicle control or test compounds during the light cycle (between
about 30 min and about 3 hours 30 min after the beginning of the
light cycle). The volume of the administered dose usually does not
exceed about 10 mL/kg.
[1777] ERG recordings are made on dark-adapted and, subsequently
(during the course of the same experiment), on light-adapted
states. For the dark-adapted response, animals are housed in a
dark-adapted environment for at least about 1 hour prior to the
recording, commencing at least about 30 minutes after the start of
the light cycle.
[1778] At about eighteen and about sixty six hours after dosing,
the mice are anesthetized with a mixture of Ketamine and Xylazine
(100 mg/kg and 20 mg/kg, respectively) and placed on a heating pad
to maintain stable core body temperature during the course of the
experiment. Pupils are dilated by placing a 5 microliter drop of
mydriatic solution (tropicamide 0.5%) in the recorded eye. A mouse
corneal monopolar contact lens electrode (Mayo Corporation,
Inazawa, Aichi, Japan) is placed on the cornea, and a subcutaneous
reference low profile needle 12 mm electrode (Grass Telefactor, W
Warwick, R.I.) is placed medial from the eye. A ground needle
electrode is placed in the tail. Data collection is obtained using
an Espion E.sup.2 (Diagnosys LLC, Littleton, Mass.) ERG recording
system with Color Dome Ganzfeld stimulator. Full dark-adapted
intensity-response function is determined following a brief white
flash stimuli of about 14 intensities ranging from about 0.0001
cds/m.sup.2 to about 333 cds/m.sup.2. Subsequently, full
light-adapted intensity-response function is determined following a
brief white flash stimuli of about 9 intensities ranging from about
0.33 cds/m.sup.2 to about 333 cds/m.sup.2. Analysis of the obtained
responses is done off-line. Intensity-response function
determination is done by fitting a sigmoid function to the data
(Naka K I, Rushton Wash., 1966; Naka K I, Rushton Wash., 1967). It
is expected that compounds of the present disclosure will depress
or suppress the dark-adapted ERG responses (measured at about 0.01
cds/m.sup.2) while minimally affecting the photopic, light-adapted
V.sub.max values when compared to control compounds.
Example 139
Effect of a Compound on Recovery of Rod B-Wave Response after Light
Bleach
[1779] ERG studies with a test compound that is a visual cycle
modulator will examine the recovery of scotopic, rod-dominated
b-wave response (measured 0 to 30 minutes with white flash stimuli
at about 0.01 cds/m.sup.2) in Balb/c mice after photo-bleach (60
cds/m.sup.2, 45 seconds) as a biomarker for suppression of rod
activity. The recovery curve at different times after single oral
dosing with 0.3 mg/kg compound is compared to vehicle. The slope of
the scotopic rod ERG b-wave recovery curve (0-30 minutes) is
calculated by linear regression and normalized to the vehicle
group. The effect on rod ERG recovery varies with time after
dosing, the greatest effect is expected to be observed at 8 hours,
and returning to near vehicle control levels at 24 hours. The
effects on ERG recovery of a range of compound doses (0.03, 0.1,
0.3 and 1 mg/kg, by oral gavage) are also studied at the 8 hour
interval. The effect of the compound on rod ERG is calculated by
linear regression as above and is expected to be dose
dependent.
Example 140
Effect of a Compound on Reduction of Lipofuscin Fluorophores
[1780] This example describes testing the capability of a test
compound to reduce the level of existing bis-retinoid,
N-retinylidene-N-retinylethanolamine (A2E) and lipofuscin
fluorophores in the retina of mice as well as prevention of the
formation of A2E and lipofuscin fluorophores. A2E is the major
fluorophore of toxic lipofuscin in ocular tissues.
[1781] The eyes of abca4-null (abca4-/-) mutant mice (see, e.g.,
Weng et al., Cell 98:13-23 (1999) have an increased accumulation of
lipofuscin fluorophores, such as A2E (see, e.g., Karan et al.,
Proc. Natl. Acad. Sci. USA 102:4164-69 (2005)). Compounds (about 1
mg/kg) or vehicle are administered daily for about three months by
oral gavage to abca4.sup.-/- mice that are about 2 months old. Mice
are sacrificed after about three months of treatment. Retinas and
RPE are extracted for A2E analysis.
[1782] A similar experiment is performed with aged balb/c mice (at
least about 10 months old). The test mice are treated with about 1
mg/kg/day of compounds for about three months and the control mice
are treated with vehicle.
[1783] Briefly, under dim red light, each pair of eye balls are
harvested, homogenized in a mixture of PBS buffer and methanol and
the A2E extracted into chloroform. The samples are dried down and
reconstituted in a water/acetonitrile mix for HPLC analysis. The
amount of A2E present is determined by comparison of the area under
the curve (AUC) of the A2E peak in the sample with an A2E
concentration/AUC curve for an A2E reference standard measuring at
440 nm.
[1784] It is expected that A2E levels are reduced upon treatment
with one or more compounds disclosed herein.
Example 141
Effect of a Compound on Retinoid Nuclear Receptor Activity
[1785] Retinoid nuclear receptor activity is associated with
transduction of the non-visual physiologic, pharmacologic, and
toxicologic retinoid signals that affect tissue and organ growth,
development, differentiation, and homeostasis.
[1786] The effect of one or more compounds disclosed herein and the
effect of a retinoic acid receptor (RAR) agonist
(E-4-[2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthylenyl)-1-propeny-
l]benzoic acid) (TTNPB), and of all-trans-retinoic acid (at-RA),
which is an RAR and retinoid X receptor (RXR) agonist, are studied
on RAR and RXR receptors essentially as described by Achkar et al.
(Proc. Natl. Acad. Sci. USA 93:4879-84 (1996)). It is expected that
the compounds of the present disclosure do not show significant
effects on retinoid nuclear receptors (RAR and RXR). By contrast,
TTNPB and at-RA activated the RXR.sub..alpha., RAR.sub..alpha.,
RAR.sub..beta. and RAR.sub..gamma. receptors as expected (Table
5).
TABLE-US-00008 TABLE 5 Com- RAR.alpha. RAR.beta. RAR.gamma.
RXR.alpha. pound EC.sub.50 (nM) EC.sub.50 (nM) EC.sub.50 (nM)
EC.sub.50 (nM) TTNPB 5.5 +/- 4.5 0.3 +/- 0.1 0.065 +/- 0.005 N/A
at-RA N/A N/A N/A 316 +/- 57 N/A = Not applicable
[1787] When ranges are used herein for physical properties, such as
molecular weight, or chemical properties, such as chemical
formulae, all combinations and subcombinations of ranges and
specific embodiments therein are intended to be included.
[1788] The various embodiments described herein can be combined to
provide further embodiments. All U.S. patents, U.S. patent
application publications, U.S. patent applications, foreign
patents, foreign patent applications, and non-patent publications
referred to in this specification and/or listed in the Application
Data Sheet, are incorporated herein by reference in their
entireties.
[1789] From the foregoing it will be appreciated that, although
specific embodiments have been described herein for purposes of
illustration, various modifications may be made. Those skilled in
the art will recognize, or be able to ascertain, using no more than
routine experimentation, many equivalents to the specific
embodiments described herein. Such equivalents are intended to be
encompassed by the following claims. In general, in the following
claims, the terms used should not be construed to limit the claims
to the specific embodiments disclosed in the specification and the
claims, but should be construed to include all possible embodiments
along with the full scope of equivalents to which such claims are
entitled. Accordingly, the claims are not limited by the
disclosure.
[1790] While preferred embodiments of the present invention have
been shown and described herein, it will be obvious to those
skilled in the art that such embodiments are provided by way of
example only. Numerous variations, changes, and substitutions will
now occur to those skilled in the art without departing from the
invention. It should be understood that various alternatives to the
embodiments of the invention described herein may be employed in
practicing the invention. It is intended that the following claims
define the scope of the invention and that methods and structures
within the scope of these claims and their equivalents be covered
thereby.
* * * * *