U.S. patent application number 14/532791 was filed with the patent office on 2015-04-30 for modulators of atp-binding cassette transporters.
The applicant listed for this patent is VERTEX PHARMACEUTICALS INCORPORATED. Invention is credited to Brian Bear, Peter D.J. Grootenhuis, Sara S. Hadida Ruah, Jason McCartney, Mark T. Miller, Mehdi Michel Jamel Numa, Fredrick Van Goor, Xiaoqing Yang, Jinglan Zhou.
Application Number | 20150119441 14/532791 |
Document ID | / |
Family ID | 38477342 |
Filed Date | 2015-04-30 |
United States Patent
Application |
20150119441 |
Kind Code |
A1 |
Hadida Ruah; Sara S. ; et
al. |
April 30, 2015 |
Modulators of ATP-Binding Cassette Transporters
Abstract
Compounds of the present invention and pharmaceutically
acceptable compositions thereof, are useful as modulators of
ATP-Binding Cassette ("ABC") transporters or fragments thereof,
including Cystic Fibrosis Transmembrane Conductance Regulator
("CFTR"). The present invention also relates to methods of treating
ABC transporter mediated diseases using compounds of the present
invention.
Inventors: |
Hadida Ruah; Sara S.; (La
Jolla, CA) ; Grootenhuis; Peter D.J.; (San Diego,
CA) ; Van Goor; Fredrick; (San Diego, CA) ;
Zhou; Jinglan; (San Diego, CA) ; Bear; Brian;
(Oceanside, CA) ; Miller; Mark T.; (San Diego,
CA) ; McCartney; Jason; (Cardiff By The Sea, CA)
; Numa; Mehdi Michel Jamel; (San Diego, CA) ;
Yang; Xiaoqing; (San Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
VERTEX PHARMACEUTICALS INCORPORATED |
Boston |
MA |
US |
|
|
Family ID: |
38477342 |
Appl. No.: |
14/532791 |
Filed: |
November 4, 2014 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
14058839 |
Oct 21, 2013 |
8912199 |
|
|
14532791 |
|
|
|
|
12829879 |
Jul 2, 2010 |
8598181 |
|
|
14058839 |
|
|
|
|
11786001 |
Apr 9, 2007 |
7776905 |
|
|
12829879 |
|
|
|
|
60790459 |
Apr 7, 2006 |
|
|
|
Current U.S.
Class: |
514/414 ; 435/29;
435/375; 506/10; 548/454 |
Current CPC
Class: |
A61P 21/02 20180101;
G01N 33/5008 20130101; A61P 5/06 20180101; A61P 31/00 20180101;
C07D 487/04 20130101; A61P 3/00 20180101; A61P 7/10 20180101; A61P
1/18 20180101; A61P 7/00 20180101; A61P 7/12 20180101; C07D 405/14
20130101; A61K 31/496 20130101; A61P 5/18 20180101; G01N 2500/02
20130101; A61P 13/12 20180101; A61P 3/06 20180101; A61P 7/02
20180101; A61P 27/02 20180101; C07D 403/12 20130101; A61P 3/10
20180101; G01N 2500/10 20130101; A61P 11/00 20180101; G01N 2333/705
20130101; A61P 11/08 20180101; A61P 27/04 20180101; A61P 25/28
20180101; A61P 35/00 20180101; A61P 13/02 20180101; A61P 15/00
20180101; A61P 25/14 20180101; A61P 19/08 20180101; C07D 405/12
20130101; G01N 33/6872 20130101; A61P 11/12 20180101; A61P 25/00
20180101; A61P 5/48 20180101; C07B 2200/05 20130101; A61K 31/5377
20130101; A61P 5/14 20180101; A61P 29/00 20180101; A61P 21/00
20180101; A61P 25/16 20180101; C07B 59/002 20130101; A61P 5/00
20180101; A61P 9/14 20180101; A61K 31/445 20130101; C07D 471/04
20130101; A61P 43/00 20180101 |
Class at
Publication: |
514/414 ;
548/454; 435/375; 435/29; 506/10 |
International
Class: |
C07D 405/12 20060101
C07D405/12; G01N 33/68 20060101 G01N033/68 |
Claims
1-39. (canceled)
40. A compound which is: ##STR00790##
41. A pharmaceutical composition comprising the compound as
described in claim 40 and a pharmaceutically acceptable
carrier.
42. A kit for use in measuring the activity of an ABC transporter
or a fragment thereof in a biological sample in vitro or in vivo,
comprising: (i) a composition comprising the compound according to
claim 40; and (ii) instructions for: a) contacting the composition
with the biological sample; and b) measuring activity of said ABC
transporter or a fragment thereof.
43. The kit of claim 42, further comprising instructions for: a)
contacting an additional composition with the biological sample; b)
measuring the activity of said ABC transporter or a fragment
thereof in the presence of said additional compound; and c)
comparing the activity of the ABC transporter in the presence of
the additional compound with the density of the ABC transporter in
the presence of a composition of formula (I).
44. The kit of claim 42, wherein the kit is used to measure the
density of CFTR.
45. A method of modulating ABC transporter activity comprising the
step of contacting said ABC transporter with a compound which is:
##STR00791##
46. The method of claim 45, wherein the ABC transporter is
CFTR.
47. A method of treating or lessening the severity of a disease in
a patient, wherein said disease is selected from cystic fibrosis,
hereditary emphysema, COPD, dry-eye disease, pancreatic
insufficiency or male infertility, said method comprising the step
of administering to said patient an effective amount of a compound
which is: ##STR00792##
48. The method of claim 47, wherein the disease is cystic fibrosis.
Description
CLAIM OF PRIORITY
[0001] The present patent application claims priority to U.S.
provisional patent application Ser. No. 60/790,459, filed on Apr.
7, 2006, which is hereby incorporated by reference in its
entirety.
TECHNICAL FIELD OF THE INVENTION
[0002] The present invention relates to modulators of ATP-Binding
Cassette ("ABC") transporters or fragments thereof, including
Cystic Fibrosis Transmembrane Conductance Regulator ("CFTR"),
compositions thereof and methods therewith. The present invention
also relates to methods of treating ABC transporter mediated
diseases using such modulators.
BACKGROUND OF THE INVENTION
[0003] ABC transporters are a family of membrane transporter
proteins that regulate the transport of a wide variety of
pharmacological agents, potentially toxic drugs, and xenobiotics,
as well as anions. ABC transporters are homologous membrane
proteins that bind and use cellular adenosine triphosphate (ATP)
for their specific activities. Some of these transporters were
discovered as multidrug resistance proteins (like the MDR1-P
glycoprotein, or the multidrug resistance protein, MRP1), defending
malignant cancer cells against chemotherapeutic agents. To date, 48
ABC Transporters have been identified and grouped into 7 families
based on their sequence identity and function.
[0004] ABC transporters regulate a variety of important
physiological roles within the body and provide defense against
harmful environmental compounds. Because of this, they represent
important potential drug targets for the treatment of diseases
associated with defects in the transporter, prevention of drug
transport out of the target cell, and intervention in other
diseases in which modulation of ABC transporter activity may be
beneficial.
[0005] One member of the ABC transporter family commonly associated
with disease is the cAMP/ATP-mediated anion channel, CFTR. CFTR is
expressed in a variety of cells types, including absorptive and
secretory epithelia cells, where it regulates anion flux across the
membrane, as well as the activity of other ion channels and
proteins. In epithelia cells, normal functioning of CFTR is
critical for the maintenance of electrolyte transport throughout
the body, including respiratory and digestive tissue. CFTR is
composed of approximately 1480 amino acids that encode a protein
made up of a tandem repeat of transmembrane domains, each
containing six transmembrane helices and a nucleotide binding
domain. The two transmembrane domains are linked by a large, polar,
regulatory (R)-domain with multiple phosphorylation sites that
regulate channel activity and cellular trafficking.
[0006] The gene encoding CFTR has been identified and sequenced
(See Gregory, R. J. et al. (1990) Nature 347:382-386; Rich, D. P.
et al. (1990) Nature 347:358-362), (Riordan, J. R. et al. (1989)
Science 245:1066-1073). A defect in this gene causes mutations in
CFTR resulting in Cystic Fibrosis ("CF"), the most common fatal
genetic disease in humans. Cystic Fibrosis affects approximately
one in every 2,500 infants in the United States. Within the general
United States population, up to 10 million people carry a single
copy of the defective gene without apparent ill effects. In
contrast, individuals with two copies of the CF associated gene
suffer from the debilitating and fatal effects of CF, including
chronic lung disease.
[0007] In patients with cystic fibrosis, mutations in CFTR
endogenously expressed in respiratory epithelia leads to reduced
apical anion secretion causing an imbalance in ion and fluid
transport. The resulting decrease in anion transport contributes to
enhanced mucus accumulation in the lung and the accompanying
microbial infections that ultimately cause death in CF patients. In
addition to respiratory disease, CF patients typically suffer from
gastrointestinal problems and pancreatic insufficiency that, if
left untreated, results in death. In addition, the majority of
males with cystic fibrosis are infertile and fertility is decreased
among females with cystic fibrosis. In contrast to the severe
effects of two copies of the CF associated gene, individuals with a
single copy of the CF associated gene exhibit increased resistance
to cholera and to dehydration resulting from diarrhea--perhaps
explaining the relatively high frequency of the CF gene within the
population.
[0008] Sequence analysis of the CFTR gene of CF chromosomes has
revealed a variety of disease causing mutations (Cutting, G. R. et
al. (1990) Nature 346:366-369; Dean, M. et al. (1990) Cell
61:863:870; and Kerem, B-S. et al. (1989) Science 245:1073-1080;
Kerem, B-S et al. (1990) Proc. Natl. Acad. Sci. USA 87:8447-8451).
To date, >1000 disease causing mutations in the CF gene have
been identified (http://www.genet.sickkids.on.ca/cftr/). The most
prevalent mutation is a deletion of phenylalanine at position 508
of the CFTR amino acid sequence, and is commonly referred to as
.DELTA.F508-CFTR. This mutation occurs in approximately 70% of the
cases of cystic fibrosis and is associated with a severe
disease.
[0009] The deletion of residue 508 in .DELTA.F508-CFTR prevents the
nascent protein from folding correctly. This results in the
inability of the mutant protein to exit the ER, and traffic to the
plasma membrane. As a result, the number of channels present in the
membrane is far less than observed in cells expressing wild-type
CFTR. In addition to impaired trafficking, the mutation results in
defective channel gating. Together, the reduced number of channels
in the membrane and the defective gating lead to reduced anion
transport across epithelia leading to defective ion and fluid
transport. (Quinton, P. M. (1990), FASEB J. 4: 2709-2727). Studies
have shown, however, that the reduced numbers of .DELTA.F508-CFTR
in the membrane are functional, albeit less than wild-type CFTR.
(Dalemans et al. (1991), Nature Lond. 354: 526-528; Denning et al.,
supra; Pasyk and Foskett (1995), J. Cell. Biochem. 270: 12347-50).
In addition to .DELTA.F508-CFTR, other disease causing mutations in
CFTR that result in defective trafficking, synthesis, and/or
channel gating could be up- or down-regulated to alter anion
secretion and modify disease progression and/or severity.
[0010] Although CFTR transports a variety of molecules in addition
to anions, it is clear that this role (the transport of anions)
represents one element in an important mechanism of transporting
ions and water across the epithelium. The other elements include
the epithelial Na.sup.+ channel, ENaC, Na.sup.+/2Cl.sup.-/K.sup.+
co-transporter, Na.sup.+--K.sup.+-ATPase pump and the basolateral
membrane K.sup.+ channels, that are responsible for the uptake of
chloride into the cell.
[0011] These elements work together to achieve directional
transport across the epithelium via their selective expression and
localization within the cell. Chloride absorption takes place by
the coordinated activity of ENaC and CFTR present on the apical
membrane and the Na.sup.+--K.sup.+-ATPase pump and Cl.sup.-
channels expressed on the basolateral surface of the cell.
Secondary active transport of chloride from the luminal side leads
to the accumulation of intracellular chloride, which can then
passively leave the cell via Cl.sup.- channels, resulting in a
vectorial transport. Arrangement of Na.sup.+/2Cl.sup.-/K.sup.+
co-transporter, Na.sup.+--K.sup.+-ATPase pump and the basolateral
membrane K.sup.+ channels on the basolateral surface and CFTR on
the luminal side coordinate the secretion of chloride via CFTR on
the luminal side. Because water is probably never actively
transported itself, its flow across epithelia depends on tiny
transepithelial osmotic gradients generated by the bulk flow of
sodium and chloride.
[0012] In addition to Cystic Fibrosis, modulation of CFTR activity
may be beneficial for other diseases not directly caused by
mutations in CFTR, such as secretory diseases and other protein
folding diseases mediated by CFTR. These include, but are not
limited to, chronic obstructive pulmonary disease (COPD), dry eye
disease, and Sjogren's Syndrome.
[0013] COPD is characterized by airflow limitation that is
progressive and not fully reversible. The airflow limitation is due
to mucus hypersecretion, emphysema, and bronchiolitis. Activators
of mutant or wild-type CFTR offer a potential treatment of mucus
hypersecretion and impaired mucociliary clearance that is common in
COPD. Specifically, increasing anion secretion across CFTR may
facilitate fluid transport into the airway surface liquid to
hydrate the mucus and optimized periciliary fluid viscosity. This
would lead to enhanced mucociliary clearance and a reduction in the
symptoms associated with COPD. Dry eye disease is characterized by
a decrease in tear aqueous production and abnormal tear film lipid,
protein and mucin profiles. There are many causes of dry eye, some
of which include age, Lasik eye surgery, arthritis, medications,
chemical/thermal burns, allergies, and diseases, such as Cystic
Fibrosis- and Sjogrens's syndrome. Increasing anion secretion via
CFTR would enhance fluid transport from the corneal endothelial
cells and secretory glands surrounding the eye to increase corneal
hydration. This would help to alleviate the symptoms associated
with dry eye disease. Sjogrens's syndrome is an autoimmune disease
in which the immune system attacks moisture-producing glands
throughout the body, including the eye, mouth, skin, respiratory
tissue, liver, vagina, and gut. Symptoms, include, dry eye, mouth,
and vagina, as well as lung disease. The disease is also associated
with rheumatoid arthritis, systemic lupus, systemic sclerosis, and
polymypositis/dermatomyositis. Defective protein trafficking is
believed to cause the disease, for which treatment options are
limited. Modulators of CFTR activity may hydrate the various organs
afflicted by the disease and help to elevate the associated
symptoms.
[0014] As discussed above, it is believed that the deletion of
residue 508 in .DELTA.F508-CFTR prevents the nascent protein from
folding correctly, resulting in the inability of this mutant
protein to exit the ER, and traffic to the plasma membrane. As a
result, insufficient amounts of the mature protein are present at
the plasma membrane and chloride transport within epithelial
tissues is significantly reduced. In fact, this cellular phenomenon
of defective ER processing of ABC transporters by the ER machinery
has been shown to be the underlying basis not only for CF disease,
but for a wide range of other isolated and inherited diseases. The
two ways that the ER machinery can malfunction is either by loss of
coupling to ER export of the proteins leading to degradation, or by
the ER accumulation of these defective/misfolded proteins [Aridor
M, et al., Nature Med., 5(7), pp 745-751 (1999); Shastry, B. S., et
al., Neurochem. International, 43, pp 1-7 (2003); Rutishauser, J.,
et al., Swiss Med Wkly, 132, pp 211-222 (2002); Morello, J P et
al., TIPS, 21, pp. 466-469 (2000); Bross P., et al., Human Mut.,
14, pp. 186-198 (1999)]. The diseases associated with the first
class, of ER malfunction are Cystic fibrosis (due to misfolded
.DELTA.F508-CFTR as discussed above), Hereditary emphysema (due to
al-antitrypsin; non Piz variants), Hereditary hemochromatosis,
Coagulation-Fibrinolysis deficiencies, such as Protein C
deficiency, Type 1 hereditary angioedema, Lipid processing
deficiencies, such as Familial hypercholesterolemia, Type 1
chylomicronemia, Abetalipoproteinemia, Lysosomal storage diseases,
such as I-cell disease/Pseudo-Hurler, Mucopolysaccharidoses (due to
Lysosomal processing enzymes), Sandhof/Tay-Sachs (due to
P-Hexosaminidase), Crigler-Najjar type II (due to
UDP-glucuronyl-sialyc-transferase),
Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus (due to
Insulin receptor), Laron dwarfism (due to Growth hormone receptor),
Myleoperoxidase deficiency, Primary hypoparathyroidism (due to
Preproparathyroid hormone), Melanoma (due to Tyrosinase). The
diseases associated with the latter class of ER malfunction are
Glycanosis CDG type 1, Hereditary emphysema (due to xal-Antitrypsin
(PiZ variant), Congenital hyperthyroidism, Osteogenesis imperfecta
(due to Type I, II, IV procollagen), Hereditary hypofibrinogenemia
(due to Fibrinogen), ACT deficiency (due to
.alpha.1-Antichymotrypsin), Diabetes insipidus (DI), Neurophyseal
DI (due to Vasopvessin hormone/V2-receptor), Neprogenic DI (due to
Aquaporin II), Charcot-Marie Tooth syndrome (due to Peripheral
myelin protein 22), Perlizaeus-Merzbacher disease,
neurodegenerative diseases such as Alzheimer's disease (due to
.beta.APP and presenilins), Parkinson's disease, Amyotrophic
lateral sclerosis, Progressive supranuclear plasy, Pick's disease,
several polyglutamine neurological disorders a such as Huntington,
Spinocerebullar ataxia type I, Spinal and bulbar muscular atrophy,
Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as
Spongiform encephalopathies, such as Hereditary Creutzfeldt-Jakob
disease (due to Prion protein processing defect), Fabry disease
(due to lysosomal .alpha.-galactosidase A) and Straussler-Scheinker
syndrome (due to Pip processing defect).
[0015] In addition to up-regulation of CFTR activity, reducing
anion secretion by CFTR modulators may be beneficial for the
treatment of secretory diarrheas, in which epithelial water
transport is dramatically increased as a result of secretagogue
activated chloride transport. The mechanism involves elevation of
cAMP and stimulation of CFTR.
[0016] Although there are numerous causes of diarrhea, the major
consequences of diarrheal diseases, resulting from excessive
chloride transport are common to all, and include dehydration,
acidosis, impaired growth and death.
[0017] Acute and chronic diarrheas represent a major medical
problem in many areas of the world. Diarrhea is both a significant
factor in malnutrition and the leading cause of death (5,000,000
deaths/year) in children less than five years old.
[0018] Secretory diarrheas are also a dangerous condition in
patients of acquired immunodeficiency syndrome (AIDS) and chronic
inflammatory bowel disease (IBD). 16 million travelers to
developing countries from industrialized nations every year develop
diarrhea, with the severity and number of cases of diarrhea varying
depending on the country and area of travel.
[0019] Diarrhea in barn animals and pets such as cows, pigs and
horses, sheep, goats, cats and dogs, also known as scours, is a
major cause of death in these animals. Diarrhea can result from any
major transition, such as weaning or physical movement, as well as
in response to a variety of bacterial or viral infections and
generally occurs within the first few hours of the animal's
life.
[0020] The most common diarrhea causing bacteria is enterotoxogenic
E. coli (ETEC) having the K99 pilus antigen. Common viral causes of
diarrhea include rotavirus and coronavirus. Other infectious agents
include cryptosporidium, giardia lamblia, and salmonella, among
others.
[0021] Symptoms of rotaviral infection include excretion of watery
feces, dehydration and weakness. Coronavirus causes a more severe
illness in the newborn animals, and has a higher mortality rate
than rotaviral infection. Often, however, a young animal may be
infected with more than one virus or with a combination of viral
and bacterial microorganisms at one time. This dramatically
increases the severity of the disease.
[0022] Accordingly, there is a need for modulators of an ABC
transporter activity, and compositions thereof, that can be used to
modulate the activity of the ABC transporter in the cell membrane
of a mammal.
[0023] There is a need for methods of treating ABC transporter
mediated diseases using such modulators of ABC transporter
activity.
[0024] There is a need for methods of modulating an ABC transporter
activity in an ex vivo cell membrane of a mammal.
[0025] There is a need for modulators of CFTR activity that can be
used to modulate the activity of CFTR in the cell membrane of a
mammal.
[0026] There is a need for methods of treating CFTR-mediated
diseases using such modulators of CFTR activity.
[0027] There is a need for methods of modulating CFTR activity in
an ex vivo cell membrane of a mammal.
SUMMARY OF THE INVENTION
[0028] It has now been found that compounds of this invention, and
pharmaceutically acceptable compositions thereof, are useful as
modulators of ABC transporter activity, particularly CTFR activity.
These compounds have the general formula I:
##STR00001##
or a pharmaceutically acceptable salt thereof, wherein R.sub.1,
R.sub.2, ring A, ring B, and n are defined below.
[0029] These compounds and pharmaceutically acceptable compositions
are useful for treating or lessening the severity of a variety of
diseases, disorders, or conditions, including, but not limited to,
cystic fibrosis, hereditary emphysema, hereditary hemochromatosis,
coagulation-fibrinolysis deficiencies, such as protein C
deficiency, Type I hereditary angioedema, lipid processing
deficiencies, such as familial hypercholesterolemia, Type 1
chylomicronemia, abetalipoproteinemia, lysosomal storage diseases,
such as I-cell disease/pseudo-Hurler, mucopolysaccharidoses,
Sandhof/Tay-Sachs, Crigler-Najjar type II,
polyendocrinopathy/hyperinsulemia, diabetes mellitus, laron
dwarfism, myleoperoxidase deficiency, primary hypoparathyroidism,
melanoma, glycanosis CDG type 1, hereditary emphysema, congenital
hyperthyroidism, osteogenesis imperfecta, hereditary
hypofibrinogenemia, ACT deficiency, diabetes insipidus,
neurophysiol, nephrogenic, Charcot-Marie Tooth syndrome,
Perlizaeus-Merzbacher disease, neurodegenerative diseases such as
Alzheimer's disease, Parkinson's disease, amyotrophic lateral
sclerosis, progressive supranuclear plasy, Pick's disease, several
polyglutamine neurological disorders asuch as Huntington,
spinocerebullar ataxia type 1, spinal and bulbar muscular atrophy,
dentatorubal pallidoluysian, and myotonic dystrophy, as well as
spongiform encephalopathies, such as hereditary Creutzfeldt-Jakob
disease, Fabry disease, Straussler-Scheinker syndrome, COPD,
dry-eye disease, and Sjogren's disease.
DETAILED DESCRIPTION OF THE INVENTION
I. Definitions
[0030] As used herein, the following definitions shall apply unless
otherwise indicated.
[0031] The term "ABC-transporter" as used herein means an
ABC-transporter protein or a fragment thereof comprising at least
one binding domain, wherein said protein or fragment thereof is
present in vivo or in vitro. The term "binding domain" as used
herein means a domain on the ABC-transporter that can bind to a
modulator. See, e.g., Hwang, T. C. et al., J. Gen. Physiol. (1998):
111(3), 477-90.
[0032] The term "CFTR" as used herein means cystic fibrosis
transmembrane conductance regulator or a mutation thereof capable
of regulator activity, including, but not limited to, .DELTA.F508
CFTR and G551D CFTR (see, e.g.,
http://www.genet.sickkids.on.ca/cftr/, for CFTR mutations).
[0033] The term "modulating" as used herein means increasing or
decreasing, e.g. activity, by a measurable amount. Compounds that
modulate ABC Transporter activity, such as CFTR activity, by
increasing the activity of the ABC Transporter, e.g., a CFTR anion
channel, are called agonists. Compounds that modulate ABC
Transporter activity, such as CFTR activity, by decreasing the
activity of the ABC Transporter, e.g., CFTR anion channel, are
called antagonists. An agonist interacts with an ABC Transporter,
such as CFTR anion channel, to increase the ability of the receptor
to transduce an intracellular signal in response to endogenous
ligand binding. An antagonist interacts with an ABC Transporter,
such as CFTR, and competes with the endogenous ligand(s) or
substrate(s) for binding site(s) on the receptor to decrease the
ability of the receptor to transduce an intracellular signal in
response to endogenous ligand binding.
[0034] The phrase "treating or reducing the severity of an ABC
Transporter mediated disease" refers both to treatments for
diseases that are directly caused by ABC Transporter and/or CFTR
activities and alleviation of symptoms of diseases not directly
caused by ABC Transporter and/or CFTR anion channel activities.
Examples of diseases whose symptoms may be affected by ABC
Transporter and/or CFTR activity include, but are not limited to,
Cystic fibrosis, Hereditary emphysema, Hereditary hemochromatosis,
Coagulation-Fibrinolysis deficiencies, such as Protein C
deficiency, Type I hereditary angioedema, Lipid processing
deficiencies, such as Familial hypercholesterolemia, Type 1
chylomicronemia, Abetalipoproteinemia, Lysosomal storage diseases,
such as I-cell disease/Pseudo-Hurler, Mucopolysaccharidoses,
Sandhof/Tay-Sachs, Crigler-Najjar type II,
Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus, Laron
dwarfism, Myleoperoxidase deficiency, Primary hypoparathyroidism,
Melanoma, Glycanosis CDO type 1, Hereditary emphysema, Congenital
hyperthyroidism, Osteogenesis imperfecta, Hereditary
hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI),
Neurophysiol DI, Nephrogenic DI, Charcot-Marie Tooth syndrome,
Perlizaeus-Merzbacher disease, neurodegenerative diseases such as
Alzheimer's disease, Parkinson's disease, Amyotrophic lateral
sclerosis, Progressive supranuclear plasy, Pick's disease, several
polyglutamine neurological disorders such as Huntington,
Spinocerebullar ataxia type I, Spinal and bulbar muscular atrophy,
Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as
Spongiform encephalopathies, such as Hereditary Creutzfeldt-Jakob
disease, Fabry disease, Straussler-Scheinker syndrome, COPD,
dry-eye disease, and Sjogren's disease.
[0035] For purposes of this invention, the chemical elements are
identified in accordance with the Periodic Table of the Elements,
CAS version, Handbook of Chemistry and Physics, 75th Ed.
Additionally, general principles of organic chemistry are described
in "Organic Chemistry", Thomas Sorrell, University Science Books,
Sausolito: 1999, and "March's Advanced Organic Chemistry", 5th Ed.,
Ed.: Smith, M. B. and March, J., John Wiley & Sons, New York:
2001, the entire contents of which are hereby incorporated by
reference.
[0036] As described herein, compounds of the invention may
optionally be substituted with one or more substituents, such as
are illustrated generally above, or as exemplified by particular
classes, subclasses, and species of the invention.
[0037] As used herein the term "aliphatic" encompasses the terms
alkyl, alkenyl, alkynyl, each of which being optionally substituted
as set forth below.
[0038] As used herein, an "alkyl" group refers to a saturated
aliphatic hydrocarbon group containing 1-12 (e.g., 1-8, 1-6, or
1-4) carbon atoms. An alkyl group can be straight or branched.
Examples of alkyl groups include, but are not limited to, methyl,
ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl,
n-pentyl, n-heptyl, or 2-ethylhexyl. An alkyl group can be
substituted (i.e., optionally substituted) with one or more
substituents such as halo, phospho, cycloaliphatic [e.g.,
cycloalkyl or cycloalkenyl], heterocycloaliphatic [e.g.,
heterocycloalkyl or heterocycloalkenyl], aryl, heteroaryl, alkoxy,
aroyl, heteroaroyl, acyl [e.g., (aliphatic)carbonyl,
(cycloaliphatic)carbonyl, or (heterocycloaliphatic)carbonyl],
nitro, cyano, amido [e.g., (cycloalkylalkyl)carbonylamino,
arylcarbonylamino, aralkylcarbonylamino,
(heterocycloalkyl)carbonylamino,
(heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino,
heteroaralkylcarbonylamino alkylaminocarbonyl,
cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl,
arylaminocarbonyl, or heteroarylaminocarbonyl], amino [e.g.,
aliphaticamino, cycloaliphaticamino, or heterocycloaliphaticamino],
sulfonyl [e.g., aliphatic-SO.sub.2-], sulfinyl, sulfanyl, sulfoxy,
urea, thiourea, sulfamoyl, sulfamide, oxo, carboxy, carbamoyl,
cycloaliphaticoxy, heterocycloaliphaticoxy, aryloxy, heteroaryloxy,
aralkyloxy, heteroarylalkoxy, alkoxycarbonyl, alkylcarbonyloxy, or
hydroxy. Without limitation, some examples of substituted alkyls
include carboxyalkyl (such as HOOC-alkyl, alkoxycarbonylalkyl, and
alkylcarbonyloxyalkyl), cyanoalkyl, hydroxyalkyl, alkoxyalkyl,
acylalkyl, aralkyl, (alkoxyaryl)alkyl, (sulfonylamino)alkyl (such
as (alkyl-SO.sub.2-amino)alkyl), aminoalkyl, amidoalkyl,
(cycloaliphatic)alkyl, or haloalkyl.
[0039] As used herein, an "alkenyl" group refers to an aliphatic
carbon group that contains 2-8 (e.g., 2-12, 2-6, or 2-4) carbon
atoms and at least one double bond. Like an alkyl group, an alkenyl
group can be straight or branched. Examples of an alkenyl group
include, but are not limited to allyl, isoprenyl, 2-butenyl, and
2-hexenyl. An alkenyl group can be optionally substituted with one
or more substituents such as halo, phospho, cycloaliphatic [e.g.,
cycloalkyl or cycloalkenyl], heterocycloaliphatic (e.g.,
heterocycloalkyl or heterocycloalkenyl], aryl, heteroaryl, alkoxy,
aroyl, heteroaroyl, acyl [e.g., (aliphatic)carbonyl,
(cycloaliphatic)carbonyl, or (heterocycloaliphatic)carbonyl],
nitro, cyano, amido [e.g., (cycloalkylalkyl)carbonylamino,
arylcarbonylamino, aralkylcarbonylamino,
(heterocycloalkyl)carbonylamino,
(heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino,
heteroaralkylcarbonylamino alkylaminocarbonyl,
cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl,
arylaminocarbonyl, or heteroarylaminocarbonyl], amino [e.g.,
alilhaticamino, cycloaliphaticamino, heterocycloaliphaticamino, or
aliphaticsulfonylamino], sulfonyl [e.g., alkyl-SO.sub.2--,
cycloaliphatic-SO.sub.2--, or aryl-SO.sub.2-], sulfinyl, sulfanyl,
sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, carboxy,
carbamoyl, cycloaliphaticoxy, heterocycloaliphaticoxy, aryloxy,
heteroaryloxy, aralkyloxy, heteroaralkoxy, alkoxycarbonyl,
alkylcarbonyloxy, or hydroxy. Without limitation, some examples of
substituted alkenyls include cyanoalkenyl, alkoxyalkenyl,
acylalkenyl, hydroxyalkenyl, aralkenyl, (alkoxyaryl)alkenyl,
(sulfonylamino)alkenyl (such as (alkyl-SO.sub.2-amino)alkenyl),
aminoalkenyl, amidoalkenyl, (cycloaliphatic)alkenyl, or
haloalkenyl.
[0040] As used herein, an "alkynyl" group refers to an aliphatic
carbon group that contains 2-8 (e.g., 2-12, 2-6, or 2-4) carbon
atoms and has at least one triple bond. An alkynyl group can be
straight or branched. Examples of an alkynyl group include, but are
not limited to, propargyl" and butynyl. An alkynyl group can be
optionally substituted with one or more substituents such as aroyl,
heteroaroyl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy,
heteroaryloxy, aralkyloxy, nitro, carboxy, cyano, halo, hydroxy,
sulfo, mercapto, sulfanyl [e.g., aliphaticsulfanyl or
cycloaliphaticsulfanyl], sulfinyl [e.g., aliphaticsulfinyl or
cycloaliphaticsulfinyl], sulfonyl [e.g., aliphatic-SO.sub.2--,
aliphaticamino-SO.sub.2--, or cycloaliphatic-SO.sub.2-], amido
[e.g., aminocarbonyl, alkylaminocarbonyl, alkylcarbonylamino,
cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl;
cycloalkylcarbonylamino, arylaminocarbonyl, arylcarbonylamino,
aralkylcarbonylamino, (heterocycloalkyl)carbonylamino,
(cycloalkylalkyl)carbonylamino, heteroaralkylcarbonylamino,
heteroarylcarbonylamino or heteroarylaminocarbonyl], urea,
thiourea, sulfamoyl, sulfamide, alkoxycarbonyl, alkylcarbonyloxy,
cycloaliphatic, heterocycloaliphatic, aryl, heteroaryl, acyl [e.g.,
(cycloaliphatic)carbonyl or (heterocycloaliphatic)carbonyl], amino
[e.g., aliphaticamino], sulfoxy, oxo, carboxy, carbamoyl,
(cycloaliphatic)oxy, (heterocycloaliphatic)oxy, or
(heteroaryl)alkoxy.
[0041] As used herein, an "amido" encompasses both "aminocarbonyl"
and "carbonylamino". These terms when used alone or in connection
with another group refer to an amido group such as
--N(R.sup.X)--C(O)--R.sup.Y or --C(O)--N(R.sup.X).sub.2, when used
terminally, and --C(O)--N(R.sup.X)-- or --N(R.sup.X)--C(O)-- when
used internally, wherein R.sup.X and R.sup.Y are defined below.
Examples of amido groups include alkylamido (such as
alkylcarbonylamino or alkylaminocarbonyl),
(heterocycloaliphatic)amido, (heteroaralkyl)amido,
(heteroaryl)amido, (heterocycloalkyl)alkylamido, arylamido,
aralkylamido, (cycloalkyl)alkylamido, or cycloalkylamido.
[0042] As used herein, an "amino" group refers to --NR.sup.XR.sup.Y
wherein each of R.sup.X and R.sup.Y is independently hydrogen,
aliphatic, cycloaliphatic, (cycloaliphatic)aliphatic, aryl,
araliphatic, heterocycloaliphatic, (heterocycloaliphatic)aliphatic,
heteroaryl, carboxy, sulfanyl, sulfinyl, sulfonyl,
(aliphatic)carbonyl, (cycloaliphatic)carbonyl,
((cycloaliphatic)aliphatic)carbonyl, arylcarbonyl,
(araliphatic)carbonyl, (heterocycloaliphatic)carbonyl,
((heterocycloaliphatic)aliphatic)carbonyl, (heteroaryl)carbonyl, or
(heteroaraliphatic)carbonyl, each of which being defined herein and
being optionally substituted. Examples of amino groups include
alkylamino, dialkylamino, or arylamino. When the term "amino" is
not the terminal group (e.g., alkylcarbonylamino), it is
represented by --NR.sup.X--. R.sup.X has the same meaning as
defined above.
[0043] As used herein, an "aryl" group used alone or as part of a
larger moiety as in "aralkyl", "aralkoxy", or "aryloxyalkyl" refers
to monocyclic (e.g., phenyl); bicyclic (e.g., indenyl,
naphthalenyl, tetrahydronaphthyl, tetrahydroindenyl); and tricyclic
(e.g., fluorenyl tetrahydrofluorenyl, or tetrahydroanthracenyl,
anthracenyl) ring systems in which the monocyclic ring system is
aromatic or at least one of the rings in a bicyclic or tricyclic
ring system is aromatic. The bicyclic and tricyclic groups include
benzofused 2-3 membered carbocyclic rings. For example, a
benzofused group includes phenyl fused with two or more C.sub.4-8
carbocyclic moieties. An aryl is optionally substituted with one or
more substituents including aliphatic [e.g., alkyl, alkenyl, or
alkynyl]; cycloaliphatic; (cycloaliphatic)aliphatic;
heterocycloaliphatic; (heterocycloaliphatic)aliphatic; aryl;
heteroaryl; alkoxy; (cycloaliphatic)oxy; (heterocycloaliphatic)oxy;
aryloxy; heteroaryloxy; (araliphatic)oxy; (heteroaraliphatic)oxy;
aroyl; heteroaroyl; amino; oxo (on a non-aromatic carbocyclic ring
of a benzofused bicyclic or tricyclic aryl); nitro; carboxy; amido;
acyl [e.g., (aliphatic)carbonyl; (cycloaliphatic)carbonyl;
((cycloaliphatic)aliphatic)carbonyl; (araliphatic)carbonyl;
(heterocycloaliphatic)carbonyl;
((heterocycloaliphatic)aliphatic)carbonyl; or
(heteroaraliphatic)carbonyl]; sulfonyl [e.g., aliphatic-SO.sub.2--
or amino-SO.sub.2-]; sulfinyl [e.g., aliphatic-S(O)-- or
cycloaliphatic-S(O)--]; sulfanyl [e.g., aliphatic-S-]; cyano; halo;
hydroxy; mercapto; sulfoxy; urea; thiourea; sulfamoyl; sulfamide;
or carbamoyl. Alternatively, an aryl can be unsubstituted.
[0044] Non-limiting examples of substituted aryls include haloaryl
[e.g., mono-, di (such as p,m-dihaloaryl), and (trihalo)aryl];
(carboxy)aryl [e.g., (alkoxycarbonyl)aryl,
((aralkyl)carbonyloxy)aryl, and (alkoxycarbonyl)aryl]; (amido)aryl
[e.g., (aminocarbonyl)aryl, (((alkylamino)alkyl)aminocarbonyl)aryl,
(alkylcarbonyl)aminoaryl, (arylaminocarbonyl)aryl, and
(((heteroaryl)amino)carbonyl)aryl]; aminoaryl (e.g.,
((alkylsulfonyl)amino)aryl or ((dialkyl)amino)aryl];
(cyanoalkyl)aryl; (alkoxy)aryl; (sulfamoyl)aryl [e.g.,
(aminosulfonyl)aryl]; (alkylsulfonyl)aryl; (cyano)aryl;
(hydroxyalkyl)aryl; ((alkoxy)alkyl)aryl; (hydroxy)aryl,
((carboxy)alkyl)aryl; (((dialkyl)amino)alkyl)aryl;
(nitroalkyl)aryl; (((alkylsulfonyl)amino)alkyl)aryl;
((heterocycloaliphatic)carbonyl)aryl; ((alkylsulfonyl)alkyl)aryl;
(cyanoalkyl)aryl; (hydroxyalkyl)aryl; (alkylcarbonyl)aryl;
alkylaryl; (trihaloalkyl)aryl; p-amino-m-alkoxycarbonylaryl;
p-amino-m-cyanoaryl; p-halo-m-aminoaryl; or
(m-(heterocycloaliphatic)-o-(alkyl))aryl.
[0045] As used herein, an "araliphatic" such as an "aralkyl" group
refers to an aliphatic group (e.g., a C.sub.1-4 alkyl group) that
is substituted with an aryl group. "Aliphatic," "alkyl," and "aryl"
are defined herein. An example of an araliphatic such as an aralkyl
group is benzyl.
[0046] As used herein, an "aralkyl" group refers to an alkyl group
(e.g., a C.sub.1-4 alkyl group) that is substituted with an aryl
group. Both "alkyl" and "aryl" have been defined above. An example
of an aralkyl group is benzyl. An aralkyl is optionally substituted
with one or more substituents such as aliphatic [e.g., alkyl,
alkenyl, or alkynyl, including carboxyalkyl, hydroxyalkyl, or
haloalkyl such as trifluoromethyl], cycloaliphatic [e.g.,
cycloalkyl or cycloalkenyl], (cycloalkyl)alkyl, heterocycloalkyl,
(heterocycloalkyl)alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy,
heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy,
heteroaralkyloxy, aroyl, heteroaroyl, nitro, carboxy,
alkoxycarbonyl, alkylcarbonyloxy, amido [e.g., aminocarbonyl,
alkylcarbonylamino, cycloalkylcarbonylamino,
(cycloalkylalkyl)carbonylamino, arylcarbonylamino,
aralkylcarbonylamino, (heterocycloalkyl)carbonylamino,
(heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino, or
heteroaralkylcarbonylamino], cyano, halo, hydroxy, acyl, mercapto,
alkylsulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo,
or carbamoyl.
[0047] As used herein, a "bicyclic ring system" includes 8-12
(e.g., 9, 10, or 11) membered structures that form two rings,
wherein the two rings have at least one atom in common (e.g., 2
atoms in common). Bicyclic ring systems include bicycloaliphatics
(e.g., bicycloalkyl or bicycloalkenyl), bicycloheteroaliphatics,
bicyclic aryls, and bicyclic heteroaryls.
[0048] As used herein, a "carbocycle" or "cycloaliphatic" group
encompasses a "cycloalkyl" group and a "cycloalkenyl" group, each
of which being optionally substituted as set forth below.
[0049] As used herein, a "cycloalkyl" group refers to a saturated
carbocyclic mono- or bicyclic (fused or bridged) ring of 3-10
(e.g., 5-10) carbon atoms. Examples of cycloalkyl groups include
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,
adamantyl, norbornyl, cubyl, octahydro-indenyl, decahydro-naphthyl,
bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[3.3.1]nonyl,
bicyclo[3.3.2.]decyl, bicyclo[2.2.2]octyl, adamantyl, or
((aminocarbonyl)cycloalkyl)cycloalkyl.
[0050] A "cycloalkenyl" group, as used herein, refers to a
non-aromatic carbocyclic ring of 3-10 (e.g., 4-8) carbon atoms
having one or more double bonds. Examples of cycloalkenyl groups
include cyclopentenyl, 1,4-cyclohexa-di-enyl, cycloheptenyl,
cyclooctenyl, hexahydro-indenyl, octahydro-naphthyl, cyclohexenyl,
cyclopentenyl, bicyclo[2.2.2]octenyl, or bicyclo[3.3.1]nonenyl.
[0051] A cycloalkyl or cycloalkenyl group can be optionally
substituted with one or more substituents such as phosphor,
aliphatic [e.g., alkyl, alkenyl, or alkynyl], cycloaliphatic,
(cycloaliphatic) aliphatic, heterocycloaliphatic,
(heterocycloaliphatic) aliphatic, aryl, heteroaryl, alkoxy,
(cycloaliphatic)oxy, (heterocycloaliphatic)oxy, aryloxy,
heteroaryloxy, (araliphatic)oxy, (heteroaraliphatic)oxy, aroyl,
heteroaroyl, amino, amido [e.g., (aliphatic)carbonylamino,
(cycloaliphatic)carbonylamino,
((cycloaliphatic)aliphatic)carbonylamino, (aryl)carbonylamino,
(araliphatic)carbonylamino, (heterocycloaliphatic)carbonylamino,
((heterocycloaliphatic)aliphatic)carbonylamino,
(heteroaryl)carbonylamino, or (heteroaraliphatic)carbonylamino],
nitro, carboxy [e.g., HOOC--, alkoxycarbonyl, or alkylcarbonyloxy],
acyl [e.g., (cycloaliphatic)carbonyl, ((cycloaliphatic)
aliphatic)carbonyl, (araliphatic)carbonyl,
(heterocycloaliphatic)carbonyl,
((heterocycloaliphatic)aliphatic)carbonyl, or
(heteroaraliphatic)carbonyl], cyano, halo, hydroxy, mercapto,
sulfonyl (e.g., alkyl-SO.sub.2-- and aryl-SO.sub.2-], sulfinyl
[e.g., alkyl-S(O)--], sulfanyl [e.g., alkyl-S-], sulfoxy, urea,
thiourea, sulfamoyl, sulfamide, oxo, or carbamoyl.
[0052] As used herein, the term "heterocycle" or
"heterocycloaliphatic" encompasses a heterocycloalkyl group and a
heterocycloalkenyl group, each of which being optionally
substituted as set forth below.
[0053] As used herein, a "heterocycloalkyl" group refers to a 3-10
membered mono- or bicylic (fused or bridged) (e.g., 5- to
10-membered mono- or bicyclic) saturated ring structure, in which
one or more of the ring atoms is a heteroatom (e.g., N, O, S, or
combinations thereof). Examples of a heterocycloalkyl group include
piperidyl, piperazyl, tetrahydropyranyl, tetrahydrofuryl,
1,4-dioxolanyl, 1,4-dithianyl, 1,3-dioxolanyl, oxazolidyl,
isoxazolidyl, morpholinyl, thiomorpholyl, octahydrobenzofuryl,
octahydrochromenyl, octahydrothiochromenyl, octahydroindolyl,
octahydropyrindinyl, decahydroquinolinyl,
octahydrobenzo[b]thiopheneyl, 2-oxa-bicyclo[2.2.2]octyl,
1-aza-bicyclo[2.2.2]octyl, 3-aza-bicyclo[3.2.1]octyl, and
2,6-dioxa-tricyclo[3.3.1.0.sup.3,7]nonyl. A monocyclic
heterocycloalkyl group can be fused with a phenyl moiety to form
structures, such as tetrahydroisoquinoline, which would be
categorized as heteroaryls.
[0054] A "heterocycloalkenyl" group, as used herein, refers to a
mono- or bicylic (e.g., 5- to 10-membered mono- or bicyclic)
non-aromatic ring structure having one or more double bonds, and
wherein one or more of the ring atoms is a heteroatom (e.g., N, O,
or S). Monocyclic and bicyclic heterocycloaliphatics are numbered
according to standard chemical nomenclature.
[0055] A heterocycloalkyl or heterocycloalkenyl group can be
optionally substituted with one or more substituents such as
phosphor, aliphatic [e.g., alkyl, alkenyl, or alkynyl],
cycloaliphatic, (cycloaliphatic)aliphatic, heterocycloaliphatic,
(heterocycloaliphatic)aliphatic, aryl, heteroaryl, alkoxy,
(cycloaliphatic)oxy, (heterocycloaliphatic)oxy, aryloxy,
heteroaryloxy, (araliphatic)oxy, (heteroaraliphatic)oxy, aroyl,
heteroaroyl, amino, amido [e.g., (aliphatic)carbonylamino,
(cycloaliphatic)carbonylamino, ((cycloaliphatic)
aliphatic)carbonylamino, (aryl)carbonylamino,
(araliphatic)carbonylamino, (heterocycloaliphatic)carbonylamino,
((heterocycloaliphatic) aliphatic)carbonylamino,
(heteroaryl)carbonylamino, or (heteroaraliphatic)carbonylamino],
nitro, carboxy [e.g., HOOC--, alkoxycarbonyl, or alkylcarbonyloxy],
acyl [e.g., (cycloaliphatic)carbonyl, ((cycloaliphatic)
aliphatic)carbonyl, (araliphatic)carbonyl,
(heterocycloaliphatic)carbonyl,
((heterocycloaliphatic)aliphatic)carbonyl, or
(heteroaraliphatic)carbonyl], nitro, cyano, halo, hydroxy,
mercapto, sulfonyl [e.g., alkylsulfonyl or arylsulfonyl], sulfinyl
[e.g., alkylsulfinyl], sulfanyl [e.g., alkylsulfanyl], sulfoxy,
urea, thiourea, sulfamoyl, sulfamide, oxo, or carbamoyl.
[0056] A "heteroaryl" group, as used herein, refers to i
monocyclic, bicyclic, or tricyclic ring system having 4 to 15 ring
atoms wherein one or more of the ring atoms is a heteroatom (e.g.,
N, O, S, or combinations thereof) and in which the monocyclic ring
system is aromatic or at least one of the rings in the bicyclic or
tricyclic ring systems is aromatic. A heteroaryl group includes a
benzofused ring system having 2 to 3 rings. For example, a
benzofused group includes benzo fused with one or two 4 to 8
membered heterocycloaliphatic moieties (e.g., indolizyl, indolyl,
isoindolyl, 3H-indolyl, indolinyl, benzo[b]furyl,
benzo[b]thiophenyl, quinolinyl, or isoquinolinyl). Some examples of
heteroaryl are azetidinyl, pyridyl, 1H-indazolyl, furyl, pyrrolyl,
thienyl, thiazolyl, oxazolyl, imidazolyl, tetrazolyl, benzofuryl,
isoquinolinyl, benzthiazolyl, xanthene, thioxanthene,
phenothiazine, dihydroindole, benzo[1,3]dioxole, benzo[b]furyl,
benzo[b]thiophenyl, indazolyl, benzimidazolyl, benzthiazolyl,
puryl, cinnolyl, quinolyl, quinazolyl, cinnolyl, phthalazyl,
quinazolyl, quinoxalyl, isoquinolyl, 4H-quinolizyl,
benzo-1,2,5-thiadiazolyl, or 1,8-naphthyridyl.
[0057] Without limitation, monocyclic heteroaryls include furyl,
thiophenyl, 2H-pyrrolyl, pyrrolyl, oxazolyl, thazolyl, imidazolyl,
pyrazolyl, isoxazolyl, isothiazolyl, 1,3,4-thiadiazolyl,
2H-pyranyl, 4-1H-pranyl, pyridyl, pyridazyl, pyrimidyl, pyrazolyl,
pyrazyl, or 1,3,5-triazyl. Monocyclic heteroaryls are numbered
according to standard chemical nomenclature.
[0058] Without limitation, bicyclic heteroaryls include indolizyl,
indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo[b]furyl,
benzo[b]thiophenyl, quinolinyl, isoquinolinyl, indolizinyl,
isoindolyl, indolyl, benzo[b]furyl, bexo[b]thiophenyl, indazolyl,
benzimidazyl, benzthiazolyl, purinyl, 4H-quinolizyl, quinolyl,
isoquinolyl, cinnolyl, phthalazyl, quinazolyl, quinoxalyl,
1,8-naphthyridyl, or pteridyl. Bicyclic heteroaryls are numbered
according to standard chemical nomenclature.
[0059] A heteroaryl is optionally substituted with one or more
substituents such as aliphatic [e.g., alkyl, alkenyl, or alkynyl];
cycloaliphatic; (cycloaliphatic)aliphatic; heterocycloaliphatic;
(heterocycloaliphatic)aliphatic; aryl; heteroaryl; alkoxy;
(cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy;
heteroaryloxy; (araliphatic)oxy; (heteroaraliphatic)oxy; aroyl;
heteroaroyl; amino; oxo (on a non-aromatic carbocyclic or
heterocyclic ring of a bicyclic or tricyclic heteroaryl); carboxy;
amido; acyl [e.g., aliphaticcarbonyl; (cycloaliphatic)carbonyl;
((cycloaliphatic)aliphatic)carbonyl; (araliphatic)carbonyl;
(heterocycloaliphatic)carbonyl;
((heterocycloaliphatic)aliphatic)carbonyl; or
(heteroaraliphatic)carbonyl]; sulfonyl [e.g., aliphaticsulfonyl or
aminosulfonyl); sulfinyl [e.g., aliphaticsulfinyl]; sulfanyl [e.g.,
aliphaticsulfanyl]; nitro; cyano; halo; hydroxy; mercapto; sulfoxy;
urea; thiourea; sulfamoyl; sulfamide; or carbamoyl. Alternatively,
a heteroaryl can be unsubstituted.
[0060] Non-limiting examples of substituted heteroaryls include
(halo)heteroaryl [e.g., mono- and di-(halo)heteroaryl];
(carboxy)heteroaryl [e.g., (alkoxycarbonyl)heteroaryl];
cyanoheteroaryl; aminoheteroaryl [e.g.,
((alkylsulfonyl)amino)heteroaryl and ((dialkyl)amino)heteroaryl];
(amido)heteroaryl [e.g., aminocarbonylheteroaryl,
((alkylcarbonyl)amino)heteroaryl,
((((alkyl)amino)alkyl)aminocarbonyl)heteroaryl,
(((heteroaryl)amino)carbonyl)heteroaryl,
((heterocycloaliphatic)carbonyl)heteroaryl, and
((alkylcarbonyl)amino)heteroaryl]; (cyanoalkyl)heteroaryl;
(alkoxy)heteroaryl; (sulfamoyl)heteroaryl [e.g.,
(aminosulfonyl)heteroaryl]; (sulfonyl)heteroaryl [e.g.,
(alkylsulfonyl)heteroaryl]; (hydroxyalkyl)heteroaryl;
(alkoxyalkyl)heteroaryl; (hydroxy)heteroaryl;
((carboxy)alkyl)heteroaryl; (((dialkyl)amino)alkyl]heteroaryl;
(heterocycloaliphatic)heteroaryl; (cycloaliphatic)heteroaryl;
(nitroalkyl)heteroaryl; (((alkylsulfonyl)amino)alkyl)heteroaryl;
((alkylsulfonyl)alkyl)heteroaryl; (cyanoalkyl)heteroaryl;
(acyl)heteroaryl [e.g., (alkylcarbonyl)heteroaryl];
(alkyl)heteroaryl, and (haloalkyl)heteroaryl [e.g.,
trihaloalkylheteroaryl].
[0061] A "heteroaraliphatic" (such as a heteroaralkyl group) as
used herein, refers to an aliphatic group (e.g., a C.sub.1-4 alkyl
group) that is substituted with a heteroaryl group. "Aliphatic,"
"alkyl," and "heteroaryl" have been defined above.
[0062] A "heteroaralkyl" group, as used herein, refers to an alkyl
group (e.g., a C.sub.1-4 alkyl group) that is substituted with a
heteroaryl group. Both "alkyl" and "heteroaryl" have been defined
above. A heteroaralkyl is optionally substituted with one or more
substituents such as alkyl (including carboxyalkyl, hydroxyalkyl,
and haloalkyl such as trifluoromethyl), alkenyl, alkynyl,
cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl,
(heterocycloalkyl)alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy,
heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy,
heteroaralkyloxy, aroyl, heteroaroyl, nitro, carboxy,
alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl,
alkylcarbonylamino, cycloalkylcarbonylamino,
(cycloalkylalkyl)carbonylamino, arylcarbonylamino,
aralkylcarbonylamino, (heterocycloalkyl)carbonylamino,
(heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino,
heteroaralkylcarbonylamino, cyano, halo, hydroxy, acyl, mercapto,
alkylsulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo,
or carbamoyl.
[0063] As used herein, "cyclic moiety" and "cyclic group" refer to
mono-, bi-, and tricyclic ring systems including cycloaliphatic,
heterocycloaliphatic, aryl, or heteroaryl, each of which has been
previously defined.
[0064] As used herein, a "bridged bicyclic ring system" refers to a
bicyclic heterocyclicaliphatic ring system or bicyclic
cycloaliphatic ring system in which the rings are bridged. Examples
of bridged bicyclic ring systems include, but are not limited to,
adamantanyl, norbornanyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl,
bicyclo[3.3.1]nonyl, bicyclo[3.2.3]nonyl, 2-oxabicyclo[2.2.2]octyl,
1-azabicyclo[2.2.2]octyl, 3-azabicyclo[3.2.1]octyl, and
2,6-dioxa-tricyclo[3.3.1.0.sup.3,7]nonyl. A bridged bicyclic ring
system can be optionally substituted with one or more substituents
such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl
such as trifluoromethyl), alkenyl, alkynyl, cycloalkyl,
(cycloalkyl)alkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, aryl,
heteroaryl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy,
heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl,
nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl,
alkylcarbonylamino, cycloalkylcarbonylamino,
(cycloalkylalkyl)carbonylamino, arylcarbonylamino,
aralkylcarbonylamino, (heterocycloalkyl)carbonylamino,
(heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino,
heteroaralkylcarbonylamino, cyano, halo, hydroxy, acyl, mercapto,
alkylsulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo,
or carbamoyl.
[0065] As used herein, an "acyl" group refers to a formyl group or
R.sup.X--C(O)-- (such as alkyl]-C(O)--, also referred to as
"alkylcarbonyl") where R.sup.X and "alkyl" have been defined
previously. Acetyl and pivaloyl are examples of acyl groups.
[0066] As used herein, an "aroyl" or "heteroaroyl" refers to an
aryl-C(O)-- or a heteroaryl-C(O)--. The aryl and heteroaryl portion
of the aroyl or heteroaroyl is optionally substituted as previously
defined.
[0067] As used herein, an "alkoxy" group refers to an alkyl-O--
group where "alkyl" has been defined previously.
[0068] As used herein, a "carbamoyl" group refers to a group having
the structure --O--CO--NR.sup.XR.sup.Y or
--NR.sup.X--CO--O--R.sup.Z, wherein R.sup.X and R.sup.Y have been
defined above and R.sup.Z can be aliphatic, aryl, araliphatic,
heterocycloaliphatic, heteroaryl, or heteroaraliphatic.
[0069] As used herein, a "carboxy" group refers to --COOH,
--COOR.sup.X, --OC(O)H, --OC(O)R.sup.X, when used as a terminal
group; or --OC(O)-- or --C(O)O-- when used as an internal
group.
[0070] As used herein, a "haloaliphatic" group refers to an
aliphatic group substituted with 1-3 halogen. For instance, the
term haloalkyl includes the group --CF.sub.3.
[0071] As used herein, a "mercapto" group refers to --SH.
[0072] As used herein, a "sulfo" group refers to --SO.sub.3H or
--SO.sub.3R.sup.X when used terminally or --S(O).sub.3-- when used
internally.
[0073] As used herein, a "sulfamide" group refers to the structure
NR.sup.X--S(O).sub.2--NR.sup.YR.sup.Z when used terminally and
--NR.sup.X--S(O).sub.2--NR.sup.Y-- when used internally, wherein
R.sup.X, R.sup.Y, and R.sup.Z have been defined above.
[0074] As used herein, a "sulfonamide" group refers to the
structure --S(O).sub.2--NR.sup.XR.sup.Y or
--NR.sup.X--S(O).sub.2--R.sup.Z when used terminally; or
--S(O).sub.2--NR.sup.X- or --NR.sup.X--S(O).sub.2-- when used
internally, wherein R.sup.X, R.sup.Y, and R.sup.Z are defined
above.
[0075] As used herein a "sulfanyl" group refers to --S--R.sup.X
when used terminally and --S-when used internally, wherein R.sup.X
has been defined above. Examples of sulfanyls include
aliphatic-S--, cycloaliphatic-S--, aryl-S--, or the like.
[0076] As used herein a "sulfinyl" group refers to --S(O)--R.sup.X
when used terminally and --S(O)--when used internally, wherein
R.sup.X has been defined above. Exemplary sulfinyl groups include
aliphatic-S(O)--, aryl-S(O)--, (cycloaliphatic(aliphatic))-S(O)--,
cycloalkyl-S(O)--, heterocycloaliphatic-S(O)--, heteroaryl-S(O)--,
or the like.
[0077] As used herein, a "sulfonyl" group refers to
--S(O).sub.2--R.sup.X when used terminally and --S(O).sub.2-- when
used internally, wherein R.sup.X has been defined above. Exemplary
sulfonyl groups include aliphatic-S(O).sub.2--, aryl-S(O).sub.2--,
(cycloaliphatic(aliphatic))-S(O).sub.2--,
cycloaliphatic-S(O).sub.2--, heterocycloaliphatic-S(O).sub.2--,
heteroaryl-S(O).sub.2--,
(cycloaliphatic(amido(aliphatic)))-S(O).sub.2-- or the like.
[0078] As used herein, a "sulfoxy" group refers to --O--SO--R.sup.X
or --SO--O--R.sup.X, when used terminally and --O--S(O)-- or
--S(O)--O-- when used internally, where R.sup.X has been defined
above.
[0079] As used herein, a "halogen" or "halo" group refers to
fluorine, chlorine, bromine or iodine.
[0080] As used herein, an "alkoxycarbonyl," which is encompassed by
the term carboxy, used alone or in connection with another group
refers to a group such as alkyl-O--C(O)--.
[0081] As used herein, an "alkoxyalkyl" refers to an alkyl group
such as alkyl-O-alkyl-, wherein alkyl has been defined above.
[0082] As used herein, a "carbonyl" refer to --C(O)--.
[0083] As used herein, an "oxo" refers to .dbd.O.
[0084] As used herein, the term "phospho" refers to phosphinates
and phosphonates.
[0085] Examples of phosphinates and phosphonates include
--P(O)(R.sup.P).sub.2, wherein R.sup.P is aliphatic, alkoxy,
aryloxy, heteroaryloxy, (cycloaliphatic)oxy,
(heterocycloaliphatic)oxy aryl, heteroaryl, cycloaliphatic or
amino.
[0086] As used herein, an "aminoalkyl" refers to the structure
(R.sup.X).sub.2N-alkyl-.
[0087] As used herein, a "cyanoalkyl" refers to the structure
(NC)-alkyl-.
[0088] As used herein, a "urea" group refers to the structure
--NR.sup.X--CO--NR.sup.YR.sup.Z and a "thiourea" group refers to
the structure --NR.sup.X--CS--NR.sup.YR.sup.Z when used terminally
and --NR.sup.X--CO--NR.sup.Y- or --NR.sup.X--CS--NR.sup.Y-- when
used internally, wherein R.sup.X, R.sup.Y, and R.sup.Z have been
defined above.
[0089] As used herein, a "guanidine" group refers to the structure
--N.dbd.C(N(R.sup.XR.sup.Y))N(R.sup.XR.sup.Y) or
--NR.sup.X--C(--NR.sup.X)NR.sup.XR.sup.Y wherein R.sup.X and
R.sup.Y have been defined above.
[0090] As used herein, the term "amidino" group refers to the
structure --C.dbd.(NR.sup.X)N(R.sup.XR.sup.Y) wherein R.sup.X and
R.sup.Y have been defined above.
[0091] In general, the term "vicinal" refers to the placement of
substituents on a group that includes two or more carbon atoms,
wherein the substituents are attached to adjacent carbon atoms.
[0092] In general, the term "geminal" refers to the placement of
substituents on a group that includes two or more carbon atoms,
wherein the substituents are attached to the same carbon atom.
[0093] The terms "terminally" and "internally" refer to the
location of a group within a substituent. A group is terminal when
the group is present at the end of the substituent not further
bonded to the rest of the chemical structure. Carboxyalkyl, i.e.,
R.sup.XO(O)C-alkyl is an example of a carboxy group used
terminally. A group is internal when the group is present in the
middle of a substituent of the chemical structure. Alkylcarboxy
(e.g., alkyl-C(O)O-- or alkyl-OC(O)--) and alkylcarboxyaryl (e.g.,
alkyl-C(O)O-aryl- or alkyl-O(CO)-aryl-) are examples of carboxy
groups used internally.
[0094] As used herein, an "aliphatic chain" refers to a branched or
straight aliphatic group (e.g., alkyl groups, alkenyl groups, or
alkynyl groups). A straight aliphatic chain has the structure
--[CH.sub.2].sub.v--, where v is 1-12. A branched aliphatic chain
is a straight aliphatic chain that is substituted with one or more
aliphatic groups. A branched aliphatic chain has the structure
--[CQQ].sub.v- where each Q is independently a hydrogen or an
aliphatic group; however, Q shall be an aliphatic group in at least
one instance. The term aliphatic chain includes alkyl chains,
alkenyl chains, and alkynyl chains, where alkyl, alkenyl, and
alkynyl are defined above.
[0095] The phrase "optionally substituted" is used interchangeably
with the phrase "substituted or unsubstituted." As described
herein, compounds of the invention can optionally be substituted
with one or more substituents, such as are illustrated generally
above, or as exemplified by particular classes, subclasses, and
species of the invention. As described herein, the variables
R.sub.1, R.sub.2, and R.sub.3, and other variables contained in
formulae described herein encompass specific groups, such as alkyl
and aryl. Unless otherwise noted, each of the specific groups for
the variables R.sub.1, R.sub.2, and R.sub.3, and other variables
contained therein can be optionally substituted with one or more
substituents described herein. Each substituent of a specific group
is further optionally substituted with one to three of halo, cyano,
oxo, alkoxy, hydroxy, amino, nitro, aryl, cycloaliphatic,
heterocycloaliphatic, heteroaryl, haloalkyl, and alkyl. For
instance, an alkyl group can be substituted with alkylsulfanyl and
the alkylsulfanyl can be optionally substituted with one to three
of halo, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl,
haloalkyl, and alkyl. As an additional example, the cycloalkyl
portion of a (cycloalkyl)carbonylamino can be optionally
substituted with one to three of halo, cyano, alkoxy, hydroxy,
nitro, haloalkyl, and alkyl. When two alkoxy groups are bound to
the same atom or adjacent atoms, the two alkxoy groups can form a
ring together with the atom(s) to which they are bound.
[0096] In general, the term "substituted," whether preceded by the
term "optionally" or not, refers to the replacement of hydrogen
radicals in a given structure with the radical of a specified
substituent. Specific substituents are described above in the
definitions and below in the description of compounds and examples
thereof. Unless otherwise indicated, an optionally substituted
group can have a substituent at each substitutable position of the
group, and when more than one position in any given structure can
be substituted with more than one substituent selected from a
specified group, the substituent can be either the same or
different at every position. A ring substituent, such as a
heterocycloalkyl, can be bound to another ring, such as a
cycloalkyl, to form a spiro-bicyclic ring system, e.g., both rings
share one common atom. As one of ordinary skill in the art will
recognize, combinations of substituents envisioned by this
invention are those combinations that result in the formation of
stable or chemically feasible compounds.
[0097] The phrase "stable or chemically feasible," as used herein,
refers to compounds that are not substantially altered when
subjected to conditions to allow for their production, detection,
and preferably their recovery, purification, and use for one or
more of the purposes disclosed herein. In some embodiments, a
stable compound or chemically feasible compound is one that is not
substantially altered when kept at a temperature of 40.degree. C.
or less, in the absence of moisture or other chemically reactive
conditions, for at least a week.
[0098] As used herein, an "effective amount" is defined as the
amount required to confer a therapeutic effect on the treated
patient, and is typically determined based on age, surface area,
weight, and condition of the patient. The interrelationship of
dosages for animals and humans (based on milligrams per meter
squared of body surface) is described by Freireich et al., Cancer
Chemother. Rep., 50: 219 (1966). Body surface area may be
approximately determined from height and weight of the patient.
See, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley, N.Y.,
537 (1970). As used herein, "patient" refers to a mammal, including
a human.
[0099] Unless otherwise stated, structures depicted herein are also
meant to include all isomeric (e.g., enantiomeric, diastereomeric,
and geometric (or conformational)) forms of the structure; for
example, the R and S configurations for each asymmetric center, (Z)
and (E) double bond isomers, and (Z) and (E) conformational
isomers. Therefore, single stereochemical isomers as well as
enantiomeric, diastereomeric, and geometric (or conformational)
mixtures of the present compounds are within the scope of the
invention. Unless otherwise stated, all tautomeric forms of the
compounds of the invention are within the scope of the invention.
Additionally, unless otherwise stated, structures depicted herein
are also meant to include compounds that differ only in the
presence of one or more isotopically enriched atoms. For example,
compounds having the present structures except for the replacement
of hydrogen by deuterium or tritium, or the replacement of a carbon
by a .sup.13C- or .sup.14C-enriched carbon are within the scope of
this invention. Such compounds are useful, for example, as
analytical tools or probes in biological assays, or as therapeutic
agents.
[0100] Compounds of the present invention are useful modulators of
ABC transporters and are useful in the treatment of ABC transporter
mediated diseases.
II. Compounds
[0101] A. Generic Compounds
[0102] The present invention relates to compounds of formula I
useful as modulators of ABC transporter activity:
##STR00002##
or a pharmaceutically acceptable salt thereof.
[0103] R.sub.1 is --Z.sup.AR.sub.4, wherein each Z.sup.A is
independently a bond or an optionally substituted branched or
straight C.sub.1-6 aliphatic chain wherein up to two carbon units
of Z.sup.A are optionally and independently replaced by --CO--,
--CS--, --CONR.sup.A--, --CONR.sup.ANR.sup.A--, --CO.sub.2--,
--OCO--, --NR.sup.ACO.sub.2--, --O--, --NR.sup.ACONR.sup.A--,
--OCONR.sup.A, --NR.sup.ANR.sup.A--, --NRCO--, --S--, --SO--,
--SO.sub.2--, --NR.sup.A--, --SO.sub.2NR.sup.A--,
--NR.sup.ASO.sub.2--, or --NR.sup.ASO.sub.2NR.sup.A--. Each R.sub.4
is independently R.sup.A, halo, --OH, --NH.sub.2, --NO.sub.2, --CN,
or --OCF.sub.3. Each R.sup.A is independently hydrogen, an
optionally substituted aliphatic, an optionally substituted
cycloaliphatic, an optionally substituted heterocycloaliphatic, an
optionally substituted aryl, or an optionally substituted
heteroaryl.
[0104] R.sub.2 is --Z.sup.BR.sub.5, wherein each Z.sup.B is
independently a bond or an optionally substituted branched or
straight C.sub.1-6 aliphatic chain wherein up to two carbon units
of Z.sup.B are optionally and independently replaced by --CO--,
--CS--, --CONR.sup.B--, --CONR.sup.BNR.sup.B--, --CO.sub.2--,
--OCO--, --NR.sup.BCO.sub.2--, --O--, --NR.sup.BCONR.sup.B--,
--OCONR.sup.B--, --NR.sup.BNR.sup.B--, --NR.sup.BCO--, --S--,
--SO--, --SO.sub.2--, --NR.sup.B--, --SO.sub.2NR.sup.B--,
--NR.sup.BSO.sub.2--, or --NR.sup.BSO.sub.2NR.sup.B--. Each R.sub.5
is independently R.sup.B, halo, --OH, --NH.sub.2, --NO.sub.2, --CN,
--CF.sub.3, or --OCF.sub.3. Each RB is independently hydrogen, an
optionally substituted aliphatic, an optionally substituted
cycloaliphatic, an optionally substituted heterocycloaliphatic, an
optionally substituted aryl, or an optionally substituted
heteroaryl. Alternatively, any two adjacent R.sub.2 groups together
with the atoms to which they are attached form an optionally
substituted carbocycle or an optionally substituted
heterocycle.
[0105] Ring A is an optionally substituted 3-7 membered monocyclic
ring having 0-3 heteroatoms selected from N, O, and S.
[0106] Ring B is a group having formula Ia:
##STR00003##
or a pharmaceutically acceptable salt thereof, wherein p is 0-3 and
each R.sub.3 and R'.sub.3 is independently --Z.sup.CR.sub.6, where
each Z.sup.C is independently a bond or an optionally substituted
branched or straight C.sub.1-6 aliphatic chain wherein up to two
carbon units of Z.sup.C are optionally and independently replaced
by --CO--, --CS--, --CONR.sup.C--, --CONR.sup.CNR.sup.C--,
--CO.sub.2--, --OCO--, --NR.sup.CCO.sub.2--, --O--,
--NR.sup.CCONR.sup.C--, --OCONR.sup.C--, --NR.sup.CNR.sup.C--,
NR.sup.CCO--, --S--, --SO--, --SO.sub.2--, --NR--,
--SO.sub.2NR.sup.C--, --NR.sup.CSO.sub.2--, or
--NR.sup.CSO.sub.2NR.sup.C--. Each R.sub.6 is independently
R.sup.C, halo, --OH, --NH.sub.2, --NO.sub.2, --CN, or --OCF.sub.3.
Each R.sup.C is independently hydrogen, an optionally substituted
aliphatic, an optionally substituted cycloaliphatic, an optionally
substituted heterocycloaliphatic, an optionally substituted aryl,
or an optionally substituted heteroaryl. Alternatively, any two
adjacent R.sub.3 groups together with the atoms to which they are
attached form an optionally substituted carbocycle or an optionally
substituted heterocycle. Furthermore, R'.sub.3 and an adjacent
R.sub.3 group, together with the atoms to which they are attached,
form an optionally substituted heterocycle.
[0107] n is 1-3.
[0108] However, in several embodiments, when ring A is
unsubstituted cyclopentyl, n is 1, R.sub.2 is 4-chloro, and R.sub.1
is hydrogen, then ring B is not 2-(tertbutyl)indol-5-yl, or
(2,6-dichlorophenyl(carbonyl))-3-methyl-1H-indol-5-yl; and when
ring A is unsubstituted cyclopentyl, n is 0, and R.sub.1 is
hydrogen, then ring B is not
##STR00004##
[0109] B. Specific Compounds
[0110] 1. R.sub.1 Group
[0111] R.sub.1 is --Z.sup.AR.sub.4, wherein each Z.sup.A is
independently a bond or an optionally substituted branched or
straight C.sub.1-6 aliphatic chain wherein up to two carbon units
of Z.sup.A are optionally and independently replaced by --CO--,
--CS--, --CONR.sup.A--, --CONR.sup.ANR.sup.A--, --CO.sub.2--,
--OCO--, --NR.sup.ACO--, --O--, --NR.sup.ACONR.sup.A--,
--OCONR.sup.A--, --NR.sup.ANR.sup.A--, --NR.sup.ACO--, --S--,
--SO--, --SO.sub.2--, --NR.sup.A--, --SO.sub.2NR.sup.A--,
--NR.sup.ASO.sub.2--, or --NR.sup.ASO.sub.2NR.sup.A--. Each R.sub.4
is independently R.sup.A, halo, --OH, --NH.sub.2, --NO.sub.2, --CN,
or --OCF.sub.3. Each R.sup.A is independently hydrogen, an
optionally substituted aliphatic, an optionally substituted
cycloaliphatic, an optionally substituted heterocycloaliphatic, an
optionally substituted aryl, or an optionally substituted
heteroaryl.
[0112] In several embodiments, R, is --Z.sup.AR.sub.4, wherein each
Z.sup.A is independently a bond or an optionally substituted
branched or straight C.sub.1-6 aliphatic chain and each R.sub.4 is
hydrogen.
[0113] In other embodiments, R.sub.1 is --Z.sup.AR.sub.4, wherein
each Z.sup.A is a bond and each R.sub.4 is hydrogen.
[0114] 2. R.sub.2 Group
[0115] Each R.sub.2 is independently --Z.sup.BR.sub.5, wherein each
Z.sup.B is independently a bond or an optionally substituted
branched or straight C.sub.1-6 aliphatic chain wherein up to two
carbon units of Z.sup.B are optionally and independently replaced
by --CO--, --CS--, --CONR.sup.B--, --CONR.sup.BNR.sup.B--,
--CO.sub.2--, --OCO--, --NR.sup.BCO.sub.2--, --O--,
--NR.sup.BCONR.sup.B--, --OCONR.sup.B--, --NR.sup.BNR.sup.B--,
--NR.sup.BCO--, --S--, --SO--, --SO.sub.2--, --NR.sup.B--,
--SO.sub.2NR.sup.B--, --NR.sup.BSO.sub.2--, or
--NR.sup.BSO.sub.2NR.sup.B--. Each R.sub.5 is independently
R.sup.B, halo, --OH, --NH.sub.2, --NO.sub.2, --CN, --CF.sub.3, or
--OCF.sub.3. Each R.sup.B is independently hydrogen, an optionally
substituted aliphatic, an optionally substituted cycloaliphatic, an
optionally substituted heterocycloaliphatic, an optionally
substituted aryl, or an optionally substituted heteroaryl.
Alternatively, any two adjacent R.sub.2 groups together with the
atoms to which they are attached form an optionally substituted
carbocycle or an optionally substituted heterocycle, or an
optionally substituted heteroaryl.
[0116] In several embodiments, R.sub.2 is an optionally substituted
aliphatic. For example, R.sub.2 is an optionally substituted
branched or straight C.sub.1-6 aliphatic chain. In other examples,
R.sub.2 is an optionally substituted branched or straight C.sub.1-6
alkyl chain, an optionally substituted branched or straight
C.sub.2-6 alkenyl chain, or an optionally substituted branched or
straight C.sub.2-6 alkynyl chain. In alternative embodiments,
R.sub.2 is a branched or straight C.sub.1-6 aliphatic chain that is
optionally substituted with 1-3 of halo, hydroxy, cyano,
cycloaliphatic, heterocycloaliphatic, aryl, heteroaryl, or
combinations thereof. For example, R.sub.2 is a branched or
straight C.sub.1-6 alkyl that is optionally substituted with 1-3 of
halo, hydroxy, cyano, cycloaliphatic, heterocycloaliphatic, aryl,
heteroaryl, or combinations thereof. In still other examples,
R.sub.2 is a methyl, ethyl, propyl, butyl, isopropyl, or
tert-butyl, each of which is optionally substituted with 1-3 of
halo, hydroxy, cyano, aryl, heteroaryl, cycloaliphatic, or
heterocycloaliphatic. In still other examples, R.sub.2 is a methyl,
ethyl, propyl, butyl, isopropyl, or tert-butyl, each of which is
unsubstituted.
[0117] In several other embodiments, R.sub.2 is an optionally
substituted branched or straight C.sub.1-5 alkoxy. For example,
R.sub.2 is a C.sub.1-5 alkoxy that is optionally substituted with
1-3 of hydroxy, aryl, heteroaryl, cycloaliphatic,
heterocycloaliphatic, or combinations thereof. In other examples,
R.sub.2 is a methoxy, ethoxy, propoxy, butoxy, or pentoxy, each of
which is optionally substituted with 1-3 of hydroxy, aryl,
heteroaryl, cycloaliphatic, heterocycloaliphatic, or combinations
thereof.
[0118] In other embodiments, R.sub.2 is hydroxy, halo, or
cyano.
[0119] In several embodiments, R.sub.2 is --Z.sup.BR.sub.5, and
Z.sup.B is independently a bond or an optionally substituted
branched or straight C.sub.1-4 aliphatic chain wherein up to two
carbon units of Z.sup.B are optionally and independently replaced
by --C(O)--, --O--, --S--, --S(O).sub.2--, or --NH--, and R.sub.5
is RB, halo, --OH, --NH.sub.2, --NO.sub.2, --CN, --CF.sub.3, or
--OCF.sub.3, and RB is hydrogen or aryl.
[0120] In several embodiments, two adjacent R.sub.2 groups form an
optionally substituted carbocycle or an optionally substituted
heterocycle. For example, two adjacent R.sub.2 groups form an
optionally substituted carbocycle or an optionally substituted
heterocycle, either of which is fused to the phenyl of formula I,
wherein the carbocycle or heterocycle has formula Ib:
##STR00005##
[0121] Each of Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4, and Z.sub.5 is
independently a bond, --CR.sub.7R'.sub.7--, --C(O)--, --NR.sub.7--,
or --O--; each R.sub.7 is independently --Z.sup.DR.sub.8, wherein
each Z.sup.D is independently a bind or an optionally substituted
branched or straight C.sub.1-6 aliphatic chain wherein up to two
carbon units of Z.sup.D are optionally and independently replaced
by --CO--, --CS--, --CONR.sup.D--, --CO.sub.2--, --OCO--,
--NR.sup.DCO.sub.2--, --O--, --NR.sup.DCONR.sup.D--,
--OCONR.sup.D--, --NR.sup.DNR, --NR.sup.DCO--, --S--, --SO--,
--SO.sub.2--, --NR.sup.D--, --SO.sub.2NR.sup.D--,
--NR.sup.DSO.sub.2--, or --NR.sub.DSO.sub.2NR.sup.D. Each R.sub.8
is independently R.sup.D, halo, --OH, --NH.sub.2, --NO.sub.2, --CN,
--CF.sub.3, or --OCF.sub.3. Each R.sup.D is independently hydrogen,
an optionally substituted cycloaliphatic, an optionally substituted
heterocycloaliphatic, an optionally substituted aryl, or an
optionally substituted heteroaryl. Each R'.sub.7 is independently
hydrogen, optionally substituted C.sub.1-6 aliphatic, hydroxy,
halo, cyano, nitro, or combinations thereof. Alternatively, any two
adjacent R.sub.7 groups together with the atoms to which they are
attached form an optionally substituted 3-7 membered carbocyclic
ring, such as an optionally substituted cyclobutyl ring, or any two
R.sub.7 and R'.sub.7 groups together with the atom or atoms to
which they are attached form an optionally substituted 3-7 membered
carbocyclic ring or a heterocarbocyclic ring.
[0122] In several other examples, two adjacent R.sub.2 groups form
an optionally substituted carbocycle. For example, two adjacent
R.sub.2 groups form an optionally substituted 5-7 membered
carbocycle that is optionally substituted with 1-3 of halo,
hydroxy, cyano, oxo, cyano, alkoxy, alkyl, or combinations thereof.
In another example, two adjacent R.sub.2 groups form a 5-6 membered
carbocycle that is optionally substituted with 1-3 of halo,
hydroxy, cyano, oxo, cyano, alkoxy, alkyl, or combinations thereof.
In still another example, two adjacent R.sub.2 groups form an
unsubstituted 5-7 membered carbocycle.
[0123] In alternative examples, two adjacent R.sub.2 groups form an
optionally substituted heterocycle. For instance, two adjacent
R.sub.2 groups form an optionally substituted 5-7 membered
heterocycle having 1-3 heteroatoms independently selected from N,
O, and S. In several examples, two adjacent R.sub.2 groups form an
optionally substituted 5-6 membered heterocycle having 1-2 oxygen
atoms. In other examples, two adjacent R.sub.2 groups form an
unsubstituted 5-7 membered heterocycle having 1-2 oxygen atoms. In
other embodiments, two adjacent R.sub.2 groups form a ring selected
from:
##STR00006## ##STR00007## ##STR00008##
[0124] In alternative examples, two adjacent R.sub.2 groups form an
optionally substituted carbocycle or an optionally substituted
heterocycle, and a third R.sub.2 group is attached to any
chemically feasible position on the phenyl of formula I. For
instance, an optionally substituted carbocycle or an optionally
substituted heterocycle, both of which is formed by two adjacent
R.sub.2 groups; a third R.sub.2 group; and the phenyl of formula I
form a group having formula Ic:
##STR00009##
[0125] Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4, and Z.sub.5 has been
defined above in formula Ib, and R.sub.2 has been defined above in
formula I.
[0126] In several embodiments, each R.sub.2 group is independently
selected from hydrogen, halo, --OCH.sub.3, --OH, --CH.sub.2OH,
--CH.sub.3, and --OCF.sub.3, and/or two adjacent R.sub.2 groups
together with the atoms to which they are attached form
##STR00010## ##STR00011## ##STR00012##
[0127] In other embodiments, R.sub.2 is at least one selected from
hydrogen, halo, methoxy, phenylmethoxy, hydroxy, hydroxymethyl,
trifluoromethoxy, and methyl.
[0128] In some embodiments, two adjacent R.sub.2 groups, together
with the atoms to which they are attached, form
##STR00013##
[0129] 3. Ring A
[0130] Ring A is an optionally substituted 3-7 membered monocyclic
ring having 0-3 heteroatoms selected from N, O, and S.
[0131] In several embodiments, ring A is an optionally substituted
3-7 membered monocyclic cycloaliphatic. For example, ring A is a
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl,
each of which is optionally substituted with 1-3 of halo, hydroxy,
C.sub.1-5 aliphatic, or combinations thereof.
[0132] In other embodiments, ring A is an optionally substituted
3-7 membered monocyclic heterocycloaliphatic. For example, ring A
is an optionally substituted 3-7 membered monocyclic
heterocycloaliphatic having 1-2 heteroatoms independently selected
from N, O, and S. In other examples, ring A is tetrahydrofuran-yl,
tetrahydro-2H-pyranyl, pyrrolidone-yl, or piperidine-yl, each of
which is optionally substituted.
[0133] In still other examples, ring A is selected from
##STR00014## ##STR00015## ##STR00016##
[0134] Each R.sub.8 is independently --Z.sup.ER.sub.9, wherein each
Z.sup.E is independently a bond or an optionally substituted
branched or straight C.sub.1-5 aliphatic chain wherein up to two
carbon units of Z.sup.E are optionally and independently replaced
by --CO--, --CS--, --CONR.sup.E--, --CO.sub.2--, --OCO--,
--NR.sup.ECO.sub.2--, --O--, --NR.sup.ECONR.sup.E--,
--OCONR.sup.E--, --NR.sup.ENR.sup.E--, --NR.sup.ECO--, --S--,
--SO--, --SO.sub.2--, --NR.sup.E--, --SO.sub.2NR.sup.E--,
--NR.sup.ESO.sub.2--, or --NR.sup.ESO.sub.2NR.sup.E--, each R.sub.9
is independently R.sup.E, --OH, --NH.sub.2, --NO.sub.2, --CN,
--CF.sub.3, oxo, or --OCF.sub.3. Each R.sup.E is independently
hydrogen, an optionally substituted cycloaliphatic, an optionally
substituted heterocycloaliphatic, an optionally substituted aryl,
or an optionally substituted heteroaryl.
[0135] q is 0-5.
[0136] In other embodiments, ring A is one selected from
##STR00017##
[0137] In several embodiments, ring A is
##STR00018##
[0138] 4. Ring B
[0139] Ring B is a group having formula Ia:
##STR00019##
or a pharmaceutically acceptable salt thereof, wherein p is
0-3.
[0140] Each R.sub.3 and R'.sub.3 is independently --Z.sup.CR.sub.6,
where each Z.sup.C is independently a bond or an optionally
substituted branched or straight C.sub.1-6 aliphatic chain wherein
up to two carbon units of Z.sup.C are optionally and independently
replaced by --CO--, --CS--, --CONR.sup.C--, --CONR.sup.CNR.sup.C--,
--CO.sub.2--, --OCO--, --NR.sup.CCO.sub.2--, --O--,
--NR.sup.CCONR.sup.C--, --OCONR.sup.C--, --NR.sup.CNR.sup.C--,
--NR.sup.CCO--, --S--, --SO--, --SO.sub.2--, --NR.sup.C--,
--SO.sub.2NR.sup.C--, --NR.sup.CSO.sub.2--, or
--NR.sup.CSO.sub.2R.sup.C--. Each R.sub.6 is independently R.sup.C,
halo, --OH, --NH.sub.2, --NO.sub.2, --CN, or --OCF.sub.3. Each
R.sup.C is independently hydrogen, an optionally substituted
aliphatic, an optionally substituted cycloaliphatic, an optionally
substituted heterocycloaliphatic, an optionally substituted aryl,
or an optionally substituted heteroaryl. Alternatively, any two
adjacent R.sub.3 groups together with the atoms to which they are
attached form an optionally substituted carbocycle or an optionally
substituted heterocycle, or R'.sub.3 and an adjacent R.sub.3, i.e.,
attached to the 2 position of the indole of formula Ia, together
with the atoms to which they are attached form an optionally
substituted heterocycle.
[0141] In several embodiments, ring B is
##STR00020##
[0142] wherein q is 0-3 and each R.sub.20 is --Z.sup.GR.sub.21,
where each Z.sup.G is independently a bond or an optionally
substituted branched or straight C.sub.1-5 aliphatic chain wherein
up to two carbon units of Z.sup.G are optionally and independently
replaced by --CO--, --CS--, --CONR.sup.G--, --CO.sub.2--, --OCO--,
--NR.sup.GCO.sub.2--, --O--, --OCONR.sup.G--, --NR.sup.GNR.sup.G--,
--NR.sup.GCO--, --S--, --SO--, --SO.sub.2--, --NR--,
--SO.sub.2NR.sup.G--, --NR.sup.GSO.sub.2--, or
--NR.sup.GSO.sub.2NR.sup.G--. Each R.sub.21 is independently
R.sup.G, halo, --OH, --NH.sub.2, --NO.sub.2, --CN, or --OCF.sub.3.
Each R.sup.G is independently hydrogen, an optionally substituted
aliphatic, an optionally substituted cycloaliphatic, an optionally
substituted heterocycloaliphatic, an optionally substituted aryl,
or an optionally substituted heteroaryl.
[0143] For example, ring B is
##STR00021##
[0144] In several embodiments, R'.sub.3 is hydrogen and R.sub.3 is
attached to the 2, 3, 4, 5, 6, or 7 position of the indole of
formula Ia. In several other examples, R.sub.3 is attached to the 2
or 3 position of the indole of formula Ia, and R.sub.3 is
independently an optionally substituted aliphatic. For instance,
R.sub.3 is an optionally substituted acyl group. In several
instances, R.sub.3 is an optionally substituted (alkoxy)carbonyl.
In other instances, R.sub.3 is (methoxy)carbonyl, (ethoxy)carbonyl,
(propoxy)carbonyl, or (butoxy)carbonyl, each of which is optionally
substituted with 1-3 of halo, hydroxy, or combinations thereof. In
other instances, R.sub.3 is an optionally substituted
(aliphatic)carbonyl. For example, R.sub.3 is an optionally
substituted (alkyl)carbonyl that is optionally substituted with 1-3
of halo, hydroxy, or combinations thereof. In other examples,
R.sub.3 is (methyl)carbonyl, (ethyl)carbonyl, (propyl)carbonyl, or
(butyl)carbonyl, each of which is optionally substituted with 1-3
of halo, hydroxy, or combinations thereof.
[0145] In several embodiments, R.sub.3 is an optionally substituted
(cycloaliphatic)carbonyl or an optionally substituted
(heterocycloaliphatic)carbonyl. In several examples, R.sub.3 is an
optionally substituted (C.sub.3--, cycloaliphatic)carbonyl. For
example, R.sub.3 is a (cyclopropyl)carbonyl, (cyclobutyl)carbonyl,
(cyclopentyl)carbonyl, (cyclohexyl)carbonyl, or
(cycloheptyl)carbonyl, each of which is optionally substituted with
aliphatic, halo, hydroxy, nitro, cyano, or combinations thereof. In
several alternative examples, R.sub.3 is an optionally substituted
(heterocycloaliphatic)carbonyl. For example, R.sub.3 is an
optionally substituted (hcterocycloaliphatic)carbonyl having 1-3
heteroatoms independently selected from N, O, and S. In other
examples, R.sub.3 is an optionally substituted
(heterocycloaliphatic)carbonyl having 1-3 heteroatoms independently
selected from N and O. In still other examples, R.sub.3 is an
optionally substituted 4-7 membered monocyclic
(heterocycloaliphatic)carbonyl having 1-3 heteroatoms independently
selected from N and 0. Alternatively, R.sub.3 is
(piperidine-1-yl)carbonyl, (pyrrolidine-1-yl)carbonyl, or
(morpholine-4-yl)carbonyl, (piperazine-1-yl)carbonyl, each of which
is optionally substituted with 1-3 of halo, hydroxy, cyano, nitro,
or aliphatic.
[0146] In still other instances, R.sub.3 is optionally substituted
(aliphatic)amido such as (aliphatic(amino(carbonyl)) that is
attached to the 2 or 3 position on the indole ring of formula Ia.
In some embodiments, R.sub.3 is an optionally substituted
(alkyl(amino))carbonyl that is attached to the 2 or 3 position on
the indole ring of formula Ia. In other embodiments, R.sub.3 is an
optionally substituted straight or branched
(aliphatic(amino))carbonyl that is attached to the 2 or 3 position
on the indole ring of formula Ia. In several examples, R.sub.3 is
(N,N-dimethyl(amino))carbonyl, (methyl(amino))carbonyl,
(ethyl(amino))carbonyl, (propyl(amino))carbonyl,
(prop-2-yl(amino))carbonyl, (dimethyl(but-2-yl(amino)))carbonyl,
(tertbutyl(amino))carbonyl, (butyl(amino))carbonyl, each of which
is optionally substituted with 1-3 of halo, hydroxy,
cycloaliphatic, heterocycloaliphatic; aryl, heteroaryl, or
combinations thereof.
[0147] In other embodiments, R.sub.3 is an optionally substituted
(alkoxy)carbonyl. For example, R.sub.3 is (methoxy)carbonyl,
(ethoxy)carbonyl, (propoxy)carbonyl, or (butoxy)carbonyl, each of
which is optionally substituted with 1-3 of halo, hydroxy, or
combinations thereof. In several instances, R.sub.3 is an
optionally substituted straight or branched C.sub.1-6 aliphatic.
For example, R.sub.3 is an optionally substituted straight or
branched C.sub.1-6 alkyl. In other examples, R.sub.3 is
independently an optionally substituted methyl, ethyl, propyl,
butyl, isopropyl, or tertbutyl, each of which is optionally
substituted with 1-3 of halo, hydroxy, cyano, nitro, or combination
thereof. In other embodiments, R.sub.3 is an optionally substituted
C.sub.3-6 cycloaliphatic. Exemplary embodiments include
cyclopropyl, 1-methyl-cycloprop-1-yl, etc. In other examples, p is
2 and the two R.sub.3 substituents are attached to the indole of
formula Ia at the 2,4- or 2,6- or 2,7-positions. Exemplary
embodiments include 6-F, 3-(optionally substituted C.sub.1-6
aliphatic or C.sub.3-6 cycloaliphatic); 7-F-2-(-(optionally
substituted C.sub.1-6 aliphatic or C.sub.3-6 cycloaliphatic)),
4F-2-(optionally substituted C, aliphatic or C.sub.3-6
cycloaliphatic); 7-CN-2-(optionally substituted C.sub.1-6 aliphatic
or C.sub.3-6 cycloaliphatic); 7-Me-2-(optionally substituted
C.sub.1-6 aliphatic or C.sub.3-6 cycloaliphatic) and
7-OMe-2-(optionally substituted C.sub.1-6 aliphatic or C.sub.3-6
cycloaliphatic).
[0148] In several embodiments, R.sub.3 is hydrogen. In several
instances, R.sub.3 is an optionally substituted straight or
branched C.sub.1-6 aliphatic. In other embodiments, R.sub.3 is an
optionally substituted C.sub.3-6 cycloaliphatic.
[0149] In several embodiments, R.sub.3 is one selected from: --H,
--CH.sub.3, --CH.sub.2OH, --CH.sub.2CH.sub.3, --CH.sub.2CH.sub.2OH,
--CH.sub.2CH.sub.2CH.sub.3, --NH.sub.2, halo, --OCH.sub.3, --CN,
--CF.sub.3, --C(O)OCH.sub.2CH.sub.3, --S(O).sub.2CH.sub.3,
--CH.sub.2NH.sub.2, --C(O)NH.sub.2,
##STR00022## ##STR00023## ##STR00024##
[0150] In another embodiment, two adjacent R.sub.3 groups form
##STR00025##
[0151] In several embodiments, R'.sub.3 is independently
--Z.sup.CR.sub.6, where each Z.sup.C is independently a bond or an
optionally substituted branched or straight C.sub.1-6 aliphatic
chain wherein up to two carbon units of Z.sup.C are optionally and
independently replaced by --CO--, --CS--, --CONR.sup.C-,
--CONR.sup.CNR.sup.C--, --CO.sub.2--, --OCO--,
--NR.sup.CCO.sub.2--, --O--, --NR.sup.CCONR.sup.C--,
--OCONR.sup.C--, --NR.sup.CNR.sup.C--, NR.sup.CCO--, --S--, --SO--,
--SO.sub.2--, --NR.sup.C--, --SO.sub.2NR.sup.C--,
--NR.sup.CSO.sub.2--, or --NR.sup.CSO.sub.2NR.sup.C--. Each R.sub.6
is independently R.sup.C, halo, --OH, --NH.sub.2, --NO.sub.2, --CN,
or --OCF.sub.3. Each R.sup.C is independently hydrogen, an
optionally substituted aliphatic, an optionally substituted
cycloaliphatic, an optionally substituted heterocycloaliphatic, or
an optionally substituted heteroaryl. In one embodiment, each
R.sup.C is hydrogen, C.sub.1-6 aliphatic, or C.sub.3-6
cycloaliphatic, wherein either of the aliphatic or cycloaliphatic
is optionally substituted with up to 4-OH substituents. In another
embodiment, R.sup.C is hydrogen, or C.sub.1-6 alkyl optionally
substituted with up to 4-OH substituents.
[0152] For example, in many embodiments, R'.sub.3 is is
independently --Z.sup.CR.sub.6, where each Z.sup.C is independently
a bond or an optionally substituted branched or straight C.sub.1-6
aliphatic chain wherein up to two carbon units of Z.sup.C are
optionally and independently replaced by --C(O)--,
--C(O)NR.sup.C--, --C(O)O--, --NR.sup.CC(O)O--, --O--,
--NR.sup.CS(O).sub.2--, or --NR.sup.C--. Each R.sub.6 is
independently R.sup.C, --OH, or --NH.sub.2. Each R.sup.C is
independently hydrogen, an optionally substituted cycloaliphatic,
an optionally substituted heterocycloaliphatic, or an optionally
substituted heteroaryl. In one embodiment, each R.sup.C is
hydrogen, C.sub.1-6 aliphatic, or C.sub.3-6 cycloaliphatic, wherein
either of the aliphatic or cycloaliphatic is optionally substituted
with up to 4-OH substituents. In another embodiment, R.sup.C is
hydrogen, or C.sub.1-6 alkyl optionally substituted with up to 4-OH
substituents.
[0153] In other embodiments, R'.sub.3 is hydrogen or
##STR00026##
[0154] wherein R.sub.31 is H or a C.sub.1-2 aliphatic that is
optionally substituted with 1-3 of halo, --OH, or combinations
thereof. R.sub.32 is -L-R.sub.33, wherein L is a bond,
--CH.sub.2--, --CH.sub.2O--, --CH.sub.2NHS(O).sub.2--,
--CH.sub.2C(O)--, --CH.sub.2NHC(O)--, or --CH.sub.2NH--; and
R.sub.33 is hydrogen, or C.sub.1-2 aliphatic, cycloaliphatic,
heterocycloaliphatic, or heteroaryl, each of which is optionally
substituted with 1 of --OH, --NH.sub.2, or --CN. For example, in
one embodiment, R.sub.31 is hydrogen and R.sub.32 is C.sub.1-2
aliphatic optionally substituted with --OH, --NH.sub.2, or
--CN.
[0155] In several embodiments, R'.sub.3 is independently selected
from one of the following: --H, --CH.sub.3, --CH.sub.2CH.sub.3,
--C(O)CH.sub.3, --CH.sub.2CH.sub.2OH, --C(O)OCH.sub.3,
##STR00027## ##STR00028## ##STR00029## ##STR00030##
[0156] 5. n term
[0157] na is 1-3.
[0158] In several embodiments, n is 1. In other embodiments, n is
2. In still other embodiments, n is 3.
[0159] C. Exemplary Compounds of the Present Invention
[0160] Exemplary compounds of the present invention include, but
are not limited to, those illustrated in Table 1 below.
TABLE-US-00001 TABLE 1 Exemplary compounds of the present
invention. ##STR00031## 1 ##STR00032## 2 ##STR00033## 3
##STR00034## 4 ##STR00035## 5 ##STR00036## 6 ##STR00037## 7
##STR00038## 8 ##STR00039## 9 ##STR00040## 10 ##STR00041## 11
##STR00042## 12 ##STR00043## 13 ##STR00044## 14 ##STR00045## 15
##STR00046## 16 ##STR00047## 17 ##STR00048## 18 ##STR00049## 19
##STR00050## 20 ##STR00051## 21 ##STR00052## 22 ##STR00053## 23
##STR00054## 24 ##STR00055## 25 ##STR00056## 26 ##STR00057##
##STR00058## 28 ##STR00059## 29 ##STR00060## 30 ##STR00061## 31
##STR00062## 32 ##STR00063## 33 ##STR00064## 34 ##STR00065## 35
##STR00066## 36 ##STR00067## 37 ##STR00068## 38 ##STR00069## 39
##STR00070## 40 ##STR00071## 41 ##STR00072## 42 ##STR00073## 43
##STR00074## 44 ##STR00075## 45 ##STR00076## 46 ##STR00077## 47
##STR00078## 48 ##STR00079## 49 ##STR00080## 50 ##STR00081## 51
##STR00082## 52 ##STR00083## 53 ##STR00084## 54 ##STR00085## 55
##STR00086## 56 ##STR00087## 57 ##STR00088## 58 ##STR00089## 59
##STR00090## 60 ##STR00091## 61 ##STR00092## 62 ##STR00093## 63
##STR00094## 64 ##STR00095## 65 ##STR00096## 66 ##STR00097## 67
##STR00098## 68 ##STR00099## 69 ##STR00100## 70 ##STR00101## 71
##STR00102## 72 ##STR00103## 73 ##STR00104## 74 ##STR00105## 75
##STR00106## 76 ##STR00107## 77 ##STR00108## 78 ##STR00109## 79
##STR00110## 80 ##STR00111## 81 ##STR00112## 82 ##STR00113## 83
##STR00114## 84 ##STR00115## 85 ##STR00116## 86 ##STR00117## 87
##STR00118## 88 ##STR00119## 89 ##STR00120## 90 ##STR00121## 91
##STR00122## 92 ##STR00123## 93 ##STR00124## 94 ##STR00125## 95
##STR00126## 96 ##STR00127## 97 ##STR00128## 98 ##STR00129## 99
##STR00130## 100 ##STR00131## 101 ##STR00132## 102 ##STR00133## 103
##STR00134## 104 ##STR00135## 105 ##STR00136## 106 ##STR00137## 107
##STR00138## 108 ##STR00139## 109 ##STR00140## 110 ##STR00141## 111
##STR00142## 112 ##STR00143## 113 ##STR00144## 114 ##STR00145## 115
##STR00146## 116 ##STR00147## 117 ##STR00148## 118 ##STR00149## 119
##STR00150## 120 ##STR00151## 121 ##STR00152## 122 ##STR00153##
123
##STR00154## 124 ##STR00155## 125 ##STR00156## 126 ##STR00157## 127
##STR00158## 128 ##STR00159## 129 ##STR00160## 130 ##STR00161## 131
##STR00162## 132 ##STR00163## 133 ##STR00164## 134 ##STR00165## 135
##STR00166## 136 ##STR00167## 137 ##STR00168## 138 ##STR00169## 139
##STR00170## 140 ##STR00171## 141 ##STR00172## 142 ##STR00173## 143
##STR00174## 144 ##STR00175## 145 ##STR00176## 146 ##STR00177## 147
##STR00178## 148 ##STR00179## 149 ##STR00180## 150 ##STR00181## 151
##STR00182## 152 ##STR00183## 153 ##STR00184## 154 ##STR00185## 155
##STR00186## 156 ##STR00187## 157 ##STR00188## 158 ##STR00189## 159
##STR00190## 160 ##STR00191## 161 ##STR00192## 162 ##STR00193## 163
##STR00194## 164 ##STR00195## 165 ##STR00196## 166 ##STR00197## 167
##STR00198## 168 ##STR00199## 169 ##STR00200## 170 ##STR00201## 171
##STR00202## 172 ##STR00203## 173 ##STR00204## 174 ##STR00205## 175
##STR00206## 176 ##STR00207## 177 ##STR00208## 178 ##STR00209## 179
##STR00210## 180 ##STR00211## 181 ##STR00212## 182 ##STR00213## 183
##STR00214## 184 ##STR00215## 185 ##STR00216## 186 ##STR00217## 187
##STR00218## 188 ##STR00219## 189 ##STR00220## 190 ##STR00221## 191
##STR00222## 192 ##STR00223## 193 ##STR00224## 194 ##STR00225## 195
##STR00226## 196 ##STR00227## 197 ##STR00228## 198 ##STR00229## 199
##STR00230## 200 ##STR00231## 201 ##STR00232## 202 ##STR00233## 203
##STR00234## 204 ##STR00235## 205 ##STR00236## 206 ##STR00237## 207
##STR00238## 208 ##STR00239## 209 ##STR00240## 210 ##STR00241## 211
##STR00242## 212 ##STR00243## 213 ##STR00244## 214 ##STR00245## 215
##STR00246## 216 ##STR00247## 217 ##STR00248## 218 ##STR00249## 219
##STR00250## 220 ##STR00251## 221 ##STR00252## 222 ##STR00253## 223
##STR00254## 224 ##STR00255## 225 ##STR00256## 226 ##STR00257## 227
##STR00258## 228 ##STR00259## 229 ##STR00260## 230 ##STR00261## 231
##STR00262## 232 ##STR00263## 233 ##STR00264## 234 ##STR00265## 235
##STR00266## 236 ##STR00267## 237 ##STR00268## 238 ##STR00269## 239
##STR00270## 240 ##STR00271## 241 ##STR00272## 242 ##STR00273## 243
##STR00274## 244 ##STR00275## 245 ##STR00276## 246 ##STR00277## 247
##STR00278## 248
##STR00279## 249 ##STR00280## 250 ##STR00281## 251 ##STR00282## 252
##STR00283## 253 ##STR00284## 254 ##STR00285## 255 ##STR00286## 256
##STR00287## 257 ##STR00288## 258 ##STR00289## 259 ##STR00290## 260
##STR00291## 261 ##STR00292## 262 ##STR00293## 263 ##STR00294## 264
##STR00295## 265 ##STR00296## 266 ##STR00297## 267 ##STR00298## 268
##STR00299## 269 ##STR00300## 270 ##STR00301## 271 ##STR00302## 272
##STR00303## 273 ##STR00304## 274 ##STR00305## 275 ##STR00306## 276
##STR00307## 277 ##STR00308## 278 ##STR00309## 279 ##STR00310## 280
##STR00311## 281 ##STR00312## 282 ##STR00313## 283 ##STR00314## 284
##STR00315## 285 ##STR00316## 286 ##STR00317## 287 ##STR00318## 288
##STR00319## 289 ##STR00320## 290 ##STR00321## 291 ##STR00322## 292
##STR00323## 293 ##STR00324## 294 ##STR00325## 295 ##STR00326## 296
##STR00327## 297 ##STR00328## 298 ##STR00329## 299 ##STR00330## 300
##STR00331## 301 ##STR00332## 302 ##STR00333## 303 ##STR00334## 304
##STR00335## 305 ##STR00336## 306
III. Subgeneric Compounds of the Present Invention
[0161] Another aspect of the present invention provides a compound
that is useful for modulating ABC transporter activity. The
compound has formula Ic:
##STR00337##
or a pharmaceutically acceptable salt thereof.
[0162] R.sub.1, R.sub.2, and ring A are defined above in formula I,
and ring B, R.sub.3 and p are defined in formula Ia. Furthermore,
when ring A is unsubstituted cyclopentyl, n is 1, R.sub.2 is
4-chloro, and R.sub.1 is hydrogen, then ring B is not
2-(tertbutyl)indol-5-yl, or
(2,6-dichlorophenyl(carbonyl))-3-methyl-1H-indol-5-yl; and when
ring A is unsubstituted cyclopentyl, n is 0, and R.sub.1 is
hydrogen, then ring B is not
##STR00338##
[0163] Another aspect of the present invention provides a compound
that is useful for modulating ABC transporter activity. The
compound has formula Id:
##STR00339##
or a pharmaceutically acceptable salt thereof.
[0164] R.sub.1, R.sub.2, and ring A are defined above in formula I,
and ring B, R.sub.3 and p are defined in formula Ia.
[0165] However, when R.sub.1 is H, n is 0, ring A is an
unsubstituted cyclopentyl, and ring B is an indole-5-yl substituted
with 1-2 of R.sub.3, then each R.sub.3 is independently
--Z.sup.GR.sub.12, where each Z.sup.G is independently a bond or an
unsubstituted branched or straight C.sub.1-6 aliphatic chain
wherein up to two carbon units of Z.sup.G are optionally and
independently replaced by --CS--, --CONR.sup.GNR.sup.G--,
--CO.sub.2--, --OCO--, --NR.sup.GCO.sub.2--, --O--,
--NR.sup.GCONR.sup.G--, .dbd.OCONR.sup.G--, --NR.sup.GNR.sup.G--,
--S--, --SO--, --SO.sub.2--, --NR.sup.G--, --SO.sub.2NR.sup.G--,
--NR.sup.GSO.sub.2--, or --NR.sup.GSO.sub.2NR.sup.C--, each
R.sub.12 is independently R.sup.G, halo, --OH, --NH.sub.2,
--NO.sub.2, --CN, or --OCF.sub.3, and each R.sup.G is independently
hydrogen, an unsubstituted aliphatic, an optionally substituted
cycloaliphatic, an optionally substituted heterocycloaliphatic, an
unsubstituted aryl, or an optionally substituted heteroaryl; or any
two adjacent R.sub.3 groups together with the atoms to which they
are attached form an optionally substituted heterocycle.
Furthermore, when R.sub.1 is H, n is 1, R.sub.2 is 4-chloro, ring A
is an unsubstituted cyclopentyl, and ring B is an indole-5-yl
substituted with 1-2 of R.sub.3, then each R.sub.3 is independently
--Z.sup.HR.sub.22, where each Z.sup.H is independently a bond or an
unsubstituted branched or straight C.sub.1-3 aliphatic chain
wherein up to two carbon units of Z.sup.H are optionally and
independently replaced by --CS--, --CONR.sup.HNR.sup.H,
--CO.sub.2--, --OCO--, --NR.sup.HCO.sub.2--, --O--,
--NR.sup.HCONR.sup.H--, --OCONR.sup.H--, --NR.sup.HNR.sup.H--,
--S--, --SO--, --SO.sub.2--, --NR.sup.H--, --SO.sub.2NR.sup.H--,
--NR.sup.HSO.sub.2--, or --NR.sup.HSO.sub.2NR.sup.H--, each
R.sub.22 is independently R.sup.H, halo, --OH, --NH.sub.2,
--NO.sub.2, --CN, or --OCF.sub.3, and each R.sup.H is independently
hydrogen, a substituted C.sub.4 alkyl, an optionally substituted
C.sub.2-6 alkenyl, an optionally substituted C.sub.2-6 alkynyl, an
optionally substituted C.sub.4 alkenyl, an optionally substituted
C.sub.4 alkynyl, an optionally substituted cycloaliphatic, an
optionally substituted heterocycloaliphatic, an optionally
substituted heteroaryl, an unsubstituted phenyl, or a
mono-substituted phenyl, or any two adjacent R.sub.3 groups
together with the atoms to which they are attached form an
optionally substituted heterocycle.
[0166] Another aspect of the present invention provides a compound
that is useful for modulating ABC transporter activity. The
compound has formula II:
##STR00340##
or a pharmaceutically acceptable salt thereof.
[0167] R.sub.1, R.sub.2, and ring A are defined above in formula I;
R.sub.3, R'.sub.3, and p are defined above in formula Ia; and
Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4, and Z.sub.5 are defined above
in formula Ib.
[0168] Another aspect of the present invention provides a compound
that is useful for modulating ABC transporter activity. The
compound has formula IIa:
##STR00341##
or a pharmaceutically acceptable salt thereof.
[0169] R.sub.1, R.sub.2, and ring A are defined above in formula I;
R.sub.3, R'.sub.3, and p are defined above in formula Ia; and
Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4, and Z.sub.5 are defined above
in formula Ib.
[0170] Another aspect of the present invention provides a compound
that is useful for modulating ABC transporter activity. The
compound has formula IIb:
##STR00342##
or a pharmaceutically acceptable salt thereof.
[0171] R.sub.1, R.sub.2, and ring A, are defined above in formula
I; R.sub.3, R'.sub.3, and p are defined above in formula Ia; and
Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4, and Z.sub.5 are defined above
in formula Ib.
[0172] Another aspect of the present invention provides a compound
that is useful for modulating ABC transporter activity. The
compound has formula IIc:
##STR00343##
or a pharmaceutically acceptable salt thereof.
[0173] R.sub.1, R.sub.2 and n are defined above in formula I; and
R.sub.3, R'.sub.3, and p are defined in formula Ia.
[0174] Another aspect of the present invention provides a compound
that is useful for modulating ABC transporter activity. The
compound has formula IId:
##STR00344##
or a pharmaceutically acceptable salt thereof.
[0175] Both R.sub.2 groups, together with the atoms to which they
are attached form a group selected from:
##STR00345## ##STR00346## ##STR00347##
[0176] R'.sub.3 is independently selected from one of the
following: --H, --CH.sub.3, --CH.sub.2CH.sub.3, --C(O)CH.sub.3,
--CH.sub.2CH.sub.2OH, --C(O)OCH.sub.3,
##STR00348## ##STR00349##
and each R.sub.3 is independently selected from --H, --CH.sub.3,
--CH.sub.2OH, --CH.sub.2CH.sub.3, --CH.sub.2CH.sub.2OH,
--CH.sub.2CH.sub.2CH.sub.3, --NH.sub.2, halo, --OCH.sub.3, --CN,
--CF.sub.3, --C(O)OCH.sub.2CH.sub.3, --S(O).sub.2CH.sub.3,
--CH.sub.2NH.sub.2, --C(O)NH.sub.2,
##STR00350## ##STR00351## ##STR00352##
IV. Generic Synthetic Schemes
[0177] The compounds of formulae (I, Ic, Id, II, Ha, IIb, IIc, and
IId) may be readily synthesized from commercially available or
known starting materials by known methods. Exemplary synthetic
routes to produce compounds of formulae (I, Ic, Id, II, IIa, IIb,
IIc, and IId) are provided below in Schemes 1-22 below.
[0178] Preparation of the compounds of the invention is achieved by
the coupling of a ring B amine with a ring A carboxylic acid as
illustrated in Scheme 1.
##STR00353##
[0179] Referring to Scheme 1, the acid 1a may be converted to the
corresponding acid chloride 1b using thionyl chloride in the
presence of a catalystic amount of dimethylformamide. Reaction of
the acid chloride with the amine
##STR00354##
provides compounds of the invention I. Alternatively, the acid 1a
may be directly coupled to the amine using known coupling reagents
such as, for example, HATU in the presence of triethylamine.
[0180] Preparation of the acids 1a may be achieved as illustrated
in Scheme 2.
##STR00355##
[0181] Referring to Scheme 2, the nitrile 2a reacts with a suitable
bromochloroalkane in the presence of sodium hydroxide and a phase
transfer catalyst such as butyltriethylammonium chloride to provide
the intermediate 2b. Hydrolysis of the nitrile of 2b provides the
acid Ia. In some instances, isolation of the intermediate 2b is
unnecessary.
[0182] The phenylacetonitriles 2a are commercially available or may
be prepared as illustrated in Scheme 3.
##STR00356##
[0183] Referring to Scheme 3, reaction of an aryl bromide 3a with
carbon monoxide in the presence of methanol and
tetrakis(triphenylphosphine)palladium (0) provides the ester 3b.
Reduction of 3b with lithium aluminum hydride provides the alcohol
3c which is converted to the halide 3d with thionyl chloride.
Reaction of 3d with sodium cyanide provides the nitrile 2a.
[0184] Other methods of producing the nitrile 2a are illustrated in
schemes 4 and 5 below.
##STR00357## ##STR00358##
[0185] Preparation of
##STR00359##
components is illustrated in the schemes that follow. A number of
methods for preparing ring B compounds wherein ring B is an indole
have been reported. See for example Angew. Chem. 2005, 44, 606; J.
Am. Chem. Soc. 2005, 127, 5342); J. Comb. Chem. 2005, 7, 130;
Tetrahedron 2006, 62, 3439; J. Chem. Soc. Perkin Trans. 1, 2000,
1045.
[0186] One method for preparing
##STR00360##
is illustrated in Scheme 6.
##STR00361##
[0187] Referring to Scheme 6, a nitroaniline 6a is converted to the
hydrazine 6b using nitrous acid in the presence of HCl and stannous
chloride. Reaction of 6b with an aldehyde or ketone
CH.sub.3C(O)R.sub.3 provides the hydrazone 6c which on treatment
with phosphoric acid in toluene leads to a mixture of nitro indoles
6d and 6e. Catalytic hydrogenation in the presence of palladium on
carbon provides a mixture of the amino indoles 6f and 6 g which may
be separated using know methods such as, for example,
chromatography.
[0188] An alternative method is illustrated in scheme 7.
##STR00362##
##STR00363##
##STR00364##
##STR00365##
##STR00366##
##STR00367##
##STR00368##
##STR00369##
##STR00370##
##STR00371##
##STR00372##
##STR00373##
##STR00374##
##STR00375##
##STR00376##
##STR00377##
##STR00378##
##STR00379##
##STR00380## ##STR00381##
##STR00382##
##STR00383##
##STR00384##
##STR00385##
##STR00386##
##STR00387##
##STR00388##
##STR00389##
##STR00390##
##STR00391##
##STR00392##
##STR00393##
##STR00394##
##STR00395##
##STR00396##
##STR00397##
[0189] In the schemes above, the radical R employed therein is a
substituent, e.g., RW as defined hereinabove. One of skill in the
art will readily appreciate that synthetic routes suitable for
various substituents of the present invention are such that the
reaction conditions and steps employed do not modify the intended
substituents.
V. Formulations, Administrations, and Uses
[0190] Accordingly, in another aspect of the present invention,
pharmaceutically acceptable compositions are provided, wherein
these compositions comprise any of the compounds as described
herein, and optionally comprise a pharmaceutically acceptable
carrier, adjuvant or vehicle. In certain embodiments, these
compositions optionally further comprise one or more additional
therapeutic agents.
[0191] It will also be appreciated that certain of the compounds of
present invention can exist in free form for treatment, or where
appropriate, as a pharmaceutically acceptable derivative or a
prodrug thereof. According to the present invention, a
pharmaceutically acceptable derivative or a prodrug includes, but
is not limited to, pharmaceutically acceptable salts, esters, salts
of such esters, or any other adduct or derivative which upon
administration to a patient in need is capable of providing,
directly or indirectly, a compound as otherwise described herein,
or a metabolite or residue thereof.
[0192] As used herein, the term "pharmaceutically acceptable salt"
refers to those salts which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of humans
and lower animals without undue toxicity, irritation, allergic
response and the like, and are commensurate with a reasonable
benefit/risk ratio. A "pharmaceutically acceptable salt" means any
salt or salt of an ester of a compound of this invention that, upon
administration to a recipient, is capable of providing, either
directly or indirectly, a compound of this invention or an
inhibitorily active metabolite or residue thereof.
[0193] Pharmaceutically acceptable salts are well known in the art.
For example, S. M. Berge, et al. describes pharmaceutically
acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66,
1-19, incorporated herein by reference. Pharmaceutically acceptable
salts of the compounds of this invention include those derived from
suitable inorganic and organic acids and bases. Examples of
pharmaceutically acceptable, nontoxic acid addition salts are salts
of an amino group formed with inorganic acids such as hydrochloric
acid, hydrobromic acid, phosphoric acid, sulfuric acid and
perchloric acid or with organic acids such as acetic acid, oxalic
acid, maleic acid, tartaric acid, citric acid, succinic acid or
malonic acid or by using other methods used in the art such as ion
exchange. Other pharmaceutically acceptable salts include adipate,
alginate, ascorbate, aspartate, benzenesulfonate, benzoate,
bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, formate, fumarate, glucoheptonate,
glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate,
hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate,
laurate, lauryl sulfate, malate, maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oleate, oxalate, palmitate, pamoate, pectinate, persulfate,
3-phenylpropionate, phosphate, picrate, pivalate, propionate,
stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
[0194] Salts derived from appropriate bases include alkali metal,
alkaline earth metal, ammonium and N.sup.+(C.sub.1-4alkyl).sub.4
salts. This invention also envisions the quaternization of any
basic nitrogen-containing groups of the compounds disclosed herein.
Water or oil-soluble or dispersible products may be obtained by
such quaternization. Representative alkali or alkaline earth metal
salts include sodium, lithium, potassium, calcium, magnesium, and
the like. Further pharmaceutically acceptable salts include, when
appropriate, nontoxic ammonium, quaternary ammonium, and amine
cations formed using counterions such as halide, hydroxide,
carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and
aryl sulfonate.
[0195] As described above, the pharmaceutically acceptable
compositions of the present invention additionally comprise a
pharmaceutically acceptable carrier, adjuvant, or vehicle, which,
as used herein, includes any and all solvents, diluents, or other
liquid vehicle, dispersion or suspension aids, surface active
agents, isotonic agents, thickening or emulsifying agents,
preservatives, solid binders, lubricants and the like, as suited to
the particular dosage form desired. Remington's Pharmaceutical
Sciences, Sixteenth Edition, E. W. Martin (Mack Publishing Co.,
Easton, Pa., 1980) discloses various carriers used in formulating
pharmaceutically acceptable compositions and known techniques for
the preparation thereof. Except insofar as any conventional carrier
medium is incompatible with the compounds of the invention, such as
by producing any undesirable biological effect or otherwise
interacting in a deleterious manner with any other component(s) of
the pharmaceutically acceptable composition, its use is
contemplated to be within the scope of this invention. Some
examples of materials which can serve as pharmaceutically
acceptable carriers include, but are not limited to, ion
exchangers, alumina, aluminum stearate, lecithin, serum proteins,
such as human serum albumin, buffer substances such as phosphates,
glycine, sorbic acid, or potassium sorbate, partial glyceride
mixtures of saturated vegetable fatty acids, water, salts or
electrolytes, such as protamine sulfate, disodium hydrogen
phosphate, potassium hydrogen phosphate, sodium chloride, zinc
salts, colloidal silica, magnesium trisilicate, polyvinyl
pyrrolidone, polyacrylates, waxes,
polyethylene-polyoxypropylene-block polymers, wool fat, sugars such
as lactose, glucose and sucrose; starches such as corn starch and
potato starch; cellulose and its derivatives such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients such as cocoa
butter and suppository waxes; oils such as peanut oil, cottonseed
oil; safflower oil; sesame oil; olive oil; corn oil and soybean
oil; glycols; such a propylene glycol or polyethylene glycol;
esters such as ethyl oleate and ethyl laurate; agar; buffering
agents such as magnesium hydroxide and aluminum hydroxide; alginic
acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl
alcohol, and phosphate buffer solutions, as well as other non-tdxic
compatible lubricants such as sodium lauryl sulfate and magnesium
stearate, as well as coloring agents, releasing agents, coating
agents, sweetening, flavoring and perfuming agents, preservatives
and antioxidants can also be present in the composition, according
to the judgment of the formulator.
[0196] In yet another aspect, the present invention provides a
method of treating a condition, disease, or disorder implicated by
ABC transporter activity. In certain embodiments, the present
invention provides a method of treating a condition, disease, or
disorder implicated by a deficiency of ABC transporter activity,
the method comprising administering a composition comprising a
compound of formulae (I, Ic, Id, II, IIa, IIb, IIc, and IId) to a
subject, preferably a mammal, in need thereof.
[0197] In certain preferred embodiments, the present invention
provides a method of treating Cystic fibrosis, Hereditary
emphysema, Hereditary hemochromatosis, Coagulation-Fibrinolysis
deficiencies, such as Protein C deficiency, Type 1 hereditary
angioedema, Lipid processing deficiencies, such as Familial
hypercholesterolemia, Type I chylomicronemia, Abetalipoproteinemia,
Lysosomal storage diseases, such as I-cell disease/Pseudo-Hurler,
Mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II,
Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus, Laron
dwarfism, Myleoperoxidase deficiency, Primary hypoparathyroidism,
Melanoma, Glycanosis CDG type 1, Hereditary emphysema, Congenital
hyperthyroidism, Osteogenesis imperfecta, Hereditary
hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI),
Neurophyseal DI, Neprogenic DI, Charcot-Marie Tooth syndrome,
Perlizaeus-Merzbacher disease, neurodegenerative diseases such as
Alzheimer's disease, Parkinson's disease, Amyotrophic lateral
sclerosis, Progressive supranuclear plasy, Pick's disease, several
polyglutamine neurological disorders asuch as Huntington,
Spinocerebullar ataxia type I, Spinal and bulbar muscular atrophy,
Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as
Spongiform encephalopathies, such as Hereditary Creutzfeldt-Jakob
disease (due to Prion protein processing defect), Fabry disease,
Straussler-Scheinker disease, secretory diarrhea, polycystic kidney
disease, chronic obstructive pulmonary disease (COPD), dry eye
disease, and Sjogren's Syndrome, comprising the step of
administering to said mammal an effective amount of a composition
comprising a compound of formulae (I, Ic, Id, II, IIa, IIb, IIc,
and IId), or a preferred embodiment thereof as set forth above.
[0198] According to an alternative preferred embodiment, the
present invention provides a method of treating cystic fibrosis
comprising the step of administering to said mammal a composition
comprising the step of administering to said mammal an effective
amount of a composition comprising a compound of formulae (I, Ic,
Id, II, IIa, IIb, IIc, and IId), or a preferred embodiment thereof
as set forth above.
[0199] According to the invention an "effective amount" of the
compound or pharmaceutically acceptable composition is that amount
effective for treating or lessening the severity of one or more of
Cystic fibrosis, Hereditary emphysema, Hereditary hemochromatosis,
Coagulation-Fibrinolysis deficiencies, such as Protein C
deficiency, Type 1 hereditary angioedema, Lipid processing
deficiencies, such as Familial hypercholesterolemia, Type 1
chylomicronemia, Abetalipoproteinemia, Lysosomal storage diseases,
such as I-cell disease/Pseudo-Hurler, Mucopolysaccharidoses,
Sandhof/Tay-Sachs, Crigler-Najjar type II,
Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus, Laron
dwarfism, Myleoperoxidase deficiency, Primary hypoparathyroidism,
Melanoma, Glycanosis CDG type 1, Hereditary emphysema, Congenital
hyperthyroidism, Osteogenesis imperfecta, Hereditary
hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI),
Neurophyseal DI, Neprogenic DI, Charcot-Marie Tooth syndrome,
Perlizaeus-Merzbacher disease, neurodegenerative diseases such as
Alzheimer's disease, Parkinson's disease, Amyotrophic lateral
sclerosis, Progressive supranuclear plasy, Pick's disease, several
polyglutamine neurological disorders asuch as Huntington,
Spinocerebullar ataxia type I, Spinal and bulbar muscular atrophy.
Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as
Spongiform encephalopathies, such as Hereditary Creutzfeldt-Jakob
disease, Fabry disease, Straussler-Scheinker disease, secretory
diarrhea, polycystic kidney disease, chronic obstructive pulmonary
disease (COPD), dry eye disease, and Sjtlgren's Syndrome.
[0200] The compounds and compositions, according to the method of
the present invention, may be administered using any amount and any
route of administration effective for treating or lessening the
severity of one or more of Cystic fibrosis, Hereditary emphysema,
Hereditary hemochromatosis, Coagulation-Fibrinolysis deficiencies,
such as Protein C deficiency, Type I hereditary angioedema, Lipid
processing deficiencies, such as Familial hypercholesterolemia,
Type I chylomicronemia, Abetalipoproteinemia, Lysosomal storage
diseases, such as I-cell disease/Pseudo-Hurler,
Mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II,
Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus, Laron
dwarfism, Myleoperoxidase deficiency, Primary hypoparathyroidism,
Melanoma, Glycanosis CDG type 1, Hereditary emphysema, Congenital
hyperthyroidism, Osteogenesis imperfecta, Hereditary
hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI),
Neurophyseal DI, Neprogenic DI, Charcot-Marie Tooth syndrome,
Perlizaeus-Merzbacher disease, neurodegenerative diseases such as
Alzheimer's disease, Parkinson's disease, Amyotrophic lateral
sclerosis, Progressive supranuclear plasy, Pick's disease, several
polyglutamine neurological disorders asuch as Huntington,
Spinocerebullar ataxia type I, Spinal and bulbar muscular atrophy,
Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as
Spongiform encephalopathies, such as Hereditary Creutzfeldt-Jakob
disease, Fabry disease, Straussler-Scheinker disease, secretory
diarrhea, polycystic kidney disease, chronic obstructive pulmonary
disease (COPD), dry eye disease, and Sjogren's Syndrome.
[0201] The exact amount required will vary from subject to subject,
depending on the species, age, and general condition of the
subject, the severity of the infection, the particular agent, its
mode of administration, and the like. The compounds of the
invention are preferably formulated in dosage unit form for ease of
administration and uniformity of dosage. The expression "dosage
unit form" as used herein refers to a physically discrete unit of
agent appropriate for the patient to be treated. It will be
understood, however, that the total daily usage of the compounds
and compositions of the present invention will be decided by the
attending physician within the scope of sound medical judgment. The
specific effective dose level for any particular patient or
organism will depend upon a variety of factors including the
disorder being treated and the severity of the disorder; the
activity of the specific compound employed; the specific
composition employed; the age, body weight, general health, sex and
diet of the patient; the time of administration, route of
administration, and rate of excretion of the specific compound
employed; the duration of the treatment; drugs used in combination
or coincidental with the specific compound employed, and like
factors well known in the medical arts. The term "patient", as used
herein, means an animal, preferably a mammal, and most preferably a
human.
[0202] The pharmaceutically acceptable compositions of this
invention can be administered to humans and other animals orally,
rectally, parenterally, intracisternally, intravaginally,
intraperitoneally, topically (as by powders, ointments, or drops),
bucally, as an oral or nasal spray, or the like, depending on the
severity of the infection being treated. In certain embodiments,
the compounds of the invention may be administered orally or
parenterally at dosage levels of about 0.01 mg/kg to about 50 mg/kg
and preferably from about 1 mg/kg to about 25 mg/kg, of subject
body weight per day, one or more times a day, to obtain the desired
therapeutic effect.
[0203] Liquid dosage forms for oral administration include, but are
not limited to, pharmaceutically acceptable emulsions,
microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the active compounds, the liquid dosage forms may
contain inert diluents commonly used in the art such as, for
example, water or other solvents, solubilizing agents and
emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in
particular, cottonseed, groundnut, corn, germ, olive, castor, and
sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene
glycols and fatty acid esters of sorbitan, and mixtures thereof.
Besides inert diluents, the oral compositions can also include
adjuvants such as wetting agents, emulsifying and suspending
agents, sweetening, flavoring, and perfuming agents.
[0204] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution, suspension or emulsion in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, U.S.P.
and isotonic sodium chloride solution. In addition, sterile, fixed
oils are conventionally employed as a solvent or suspending medium.
For this purpose any bland fixed oil can be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid are used in the preparation of injectables.
[0205] The injectable formulations can be sterilized, for example,
by filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
[0206] In order to prolong the effect of a compound of the present
invention, it is often desirable to slow the absorption of the
compound from subcutaneous or intramuscular injection. This may be
accomplished by the use of a liquid suspension of crystalline or
amorphous material with poor water solubility. The rate of
absorption of the compound then depends upon its rate of
dissolution that, in turn, may depend upon crystal size and
crystalline form. Alternatively, delayed absorption of a
parenterally administered compound form is accomplished by
dissolving or suspending the compound in an oil vehicle. Injectable
depot forms are made by forming microencapsule matrices of the
compound in biodegradable polymers such as
polylactide-polyglycolide. Depending upon the ratio of compound to
polymer and the nature of the particular polymer employed, the rate
of compound release can be controlled. Examples of other
biodegradable polymers include poly(orthoesters) and
poly(anhydrides). Depot injectable formulations are also prepared
by entrapping the compound in liposomes or microemulsions that are
compatible with body tissues.
[0207] Compositions for rectal or vaginal administration are
preferably suppositories which can be prepared by mixing the
compounds of this invention with suitable non-irritating excipients
or carriers such as cocoa butter, polyethylene glycol or a
suppository wax which are solid at ambient temperature but liquid
at body temperature and therefore melt in the rectum or vaginal
cavity and release the active compound.
[0208] Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active compound is mixed with at least one inert,
pharmaceutically acceptable excipient or carrier such as sodium
citrate or dicalcium phosphate and/or a) fillers or extenders such
as starches, lactose, sucrose, glucose, mannitol, and silicic acid,
b) binders such as, for example, carboxymethylcellulose, alginates,
gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants
such as glycerol, d) disintegrating agents such as agar-agar,
calcium carbonate, potato or tapioca starch, alginic acid, certain
silicates, and sodium carbonate, e) solution retarding agents such
as paraffin, f) absorption accelerators such as quaternary ammonium
compounds, g) wetting agents such as, for example, cetyl alcohol
and glycerol monostearate, h) absorbents such as kaolin and
bentonite clay, and i) lubricants such as talc, calcium stearate,
magnesium stearate, solid polyethylene glycols, sodium lauryl
sulfate, and mixtures thereof. In the case of capsules, tablets and
pills, the dosage form may also comprise buffering agents.
[0209] Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like. The solid dosage forms of
tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings and other
coatings well known in the pharmaceutical formulating art. They may
optionally contain opacifying agents and can also be of a
composition that they release the active ingredient(s) only, or
preferentially, in a certain part of the intestinal tract,
optionally, in a delayed manner. Examples of embedding compositions
that can be used include polymeric substances and waxes. Solid
compositions of a similar type may also be employed as fillers in
soft and hard-filled gelatin capsules using such excipients as
lactose or milk sugar as well as high molecular weight polethylene
glycols and the like.
[0210] The active compounds can also be in microencapsulated form
with one or more excipients as noted above. The solid dosage forms
of tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings, release
controlling coatings and other coatings well known in the
pharmaceutical formulating art. In such solid dosage forms the
active compound may be admixed with at least one inert diluent such
as sucrose, lactose or starch. Such dosage forms may also comprise,
as is normal practice, additional substances other than inert
diluents, e.g., tableting lubricants and other tableting aids such
a magnesium stearate and microcrystalline cellulose. In the case of
capsules, tablets and pills, the dosage forms may also comprise
buffering agents. They may optionally contain opacifying agents and
can also be of a composition that they release the active
ingredient(s) only, or preferentially, in a certain part of the
intestinal tract, optionally, in a delayed manner. Examples of
embedding compositions that can be used include polymeric
substances and waxes.
[0211] Dosage forms for topical or transdermal administration of a
compound of this invention include ointments, pastes, creams,
lotions, gels, powders, solutions, sprays, inhalants or patches.
The active component is admixed under sterile conditions with a
pharmaceutically acceptable carrier and any needed preservatives or
buffers as may be required. Ophthalmic formulation, eardrops, and
eye drops are also contemplated as being within the scope of this
invention. Additionally, the present invention contemplates the use
of transdermal patches, which have the added advantage of providing
controlled delivery of a compound to the body. Such dosage forms
are prepared by dissolving or dispensing the compound in the proper
medium. Absorption enhancers can also be used to increase the flux
of the compound across the skin. The rate can be controlled by
either providing a rate controlling membrane or by dispersing the
compound in a polymer matrix or gel.
[0212] As described generally above, the compounds of the invention
are useful as modulators of ABC transporters. Thus, without wishing
to be bound by any particular theory, the compounds and
compositions are particularly useful for treating or lessening the
severity of a disease, condition, or disorder where hyperactivity
or inactivity of ABC transporters is implicated in the disease,
condition, or disorder. When hyperactivity or inactivity of an ABC
transporter is implicated in a particular disease, condition, or
disorder, the disease, condition, or disorder may also be referred
to as a "ABC transporter-mediated disease, condition or disorder".
Accordingly, in another aspect, the present invention provides a
method for treating or lessening the severity of a disease,
condition, or disorder where hyperactivity or inactivity of an ABC
transporter is implicated in the disease state.
[0213] The activity of a compound utilized in this invention as a
modulator of an ABC transporter may be assayed according to methods
described generally in the art and in the Examples herein.
[0214] It will also be appreciated that the compounds and
pharmaceutically acceptable compositions of the present invention
can be employed in combination therapies, that is, the compounds
and pharmaceutically acceptable compositions can be administered
concurrently with, prior to, or subsequent to, one or more other
desired therapeutics or medical procedures. The particular
combination of therapies (therapeutics or procedures) to employ in
a combination regimen will take into account compatibility of the
desired therapeutics and/or procedures and the desired therapeutic
effect to be achieved. It will also be appreciated that the
therapies employed may achieve a desired effect for the same
disorder (for example, an inventive compound may be administered
concurrently with another agent used to treat the same disorder),
or they may achieve different effects (e.g., control of any adverse
effects). As used herein, additional therapeutic agents that are
normally administered to treat or prevent a particular disease, or
condition, are known as "appropriate for the disease, or condition,
being treated".
[0215] The amount of additional therapeutic agent present in the
compositions of this invention will be no more than the amount that
would normally be administered in a composition comprising that
therapeutic agent as the only active agent. Preferably the amount
of additional therapeutic agent in the presently disclosed
compositions will range from about 50% to 100% of the amount
normally present in a composition comprising that agent as the only
therapeutically active agent.
[0216] The compounds of this invention or pharmaceutically
acceptable compositions thereof may also be incorporated into
compositions for coating an implantable medical device, such as
prostheses, artificial valves, vascular grafts, stents and
catheters. Accordingly, the present invention, in another aspect,
includes a composition for coating an implantable device comprising
a compound of the present invention as described generally above,
and in classes and subclasses herein, and a carrier suitable for
coating said implantable device. In still another aspect, the
present invention includes an implantable device coated with a
composition comprising a compound of the present invention as
described generally above, and in classes and subclasses herein,
and a carrier suitable for coating said implantable device.
Suitable coatings and the general preparation of coated implantable
devices are described in U.S. Pat. Nos. 6,099,562; 5,886,026; and
5,304,121. The coatings are typically biocompatible polymeric
materials such as a hydrogel polymer, polymethyldisiloxane,
polycaprolactone, polyethylene glycol, polylactic acid, ethylene
vinyl acetate, and mixtures thereof. The coatings may optionally be
further covered by a suitable topcoat of fluorosilicone,
polysaccarides, polyethylene glycol, phospholipids or combinations
thereof to impart controlled release characteristics in the
composition.
[0217] Another aspect of the invention relates to modulating ABC
transporter activity in a biological sample or a patient (e.g., in
vitro or in vivo), which method comprises administering to the
patient, or contacting said biological sample with a compound of
formula I or a composition comprising said compound. The term
"biological sample", as used herein, includes, without limitation,
cell cultures or extracts thereof; biopsied material obtained from
a mammal or extracts thereof; and blood, saliva, urine, feces,
semen, tears, or other body fluids or extracts thereof.
[0218] Modulation of ABC transporter activity in a biological
sample is useful for a variety of purposes that are known to one of
skill in the art. Examples of such purposes include, but are not
limited to, the study of ABC transporters in biological and
pathological phenomena; and the comparative evaluation of new
modulators of ABC transporters.
[0219] In yet another embodiment, a method of modulating activity
of an anion channel in vitro or in vivo, is provided comprising the
step of contacting said channel with a compound of formulae (I, Ic,
Id, II, IIa, IIb, IIc, and IId). In preferred embodiments, the
anion channel is a chloride channel or a bicarbonate channel. In
other preferred embodiments, the anion channel is a chloride
channel.
[0220] According to an alternative embodiment, the present
invention provides a method of increasing the number of functional
ABC transporters in a membrane of a cell, comprising the step of
contacting said cell with a compound of formulae (I, Ic, Id, II,
IIa, IIb, IIc, and IId). The term "functional ABC transporter" as
used herein means an ABC transporter that is capable of transport
activity. In preferred embodiments, said functional ABC transporter
is CFTR.
[0221] According to another preferred embodiment, the activity of
the ABC transporter is measured by measuring the transmembrane
voltage potential. Means for measuring the voltage potential across
a membrane in the biological sample may employ any of the known
methods in the art, such as optical membrane potential assay or
other electrophysiological methods.
[0222] The optical membrane potential assay utilizes
voltage-sensitive FRET sensors described by Gonzalez and Tsien
(See, Gonzalez, J. E. and R. Y. Tsien (1995) "Voltage sensing by
fluorescence resonance energy transfer in single cells" Biophys J
69(4): 1272-80, and Gonzalez, J. E. and R. Y. Tsien (1997)
"Improved indicators of cell membrane potential that use
fluorescence resonance energy transfer" Chem Biol 4(4): 269-77) in
combination with instrumentation for measuring fluorescence changes
such as the Voltage/Ion Probe Reader (VIPR) (See, Gonzalez, J. E.,
K. Oades, et al. (1999) "Cell-based assays and instrumentation for
screening ion-channel targets" Drug Discov Today 4(9):
431-439).
[0223] These voltage sensitive assays are based on the change in
fluorescence resonant energy transfer (FRET) between the
membrane-soluble, voltage-sensitive dye, DiSBAC.sub.2(3), and a
fluorescent phospholipid, CC2-DMPE, which is attached to the outer
leaflet of the plasma membrane and acts as a FRET donor. Changes in
membrane potential (V.sub.m) cause the negatively charged
DiSBAC.sub.2(3) to redistribute across the plasma membrane and the
amount of energy transfer from CC2-DMPE changes accordingly. The
changes in fluorescence emission can be monitored using VIPR.TM.
II, which is an integrated liquid handler and fluorescent detector
designed to conduct cell-based screens in 96- or 384-well
microtiter plates.
[0224] In another aspect the present invention provides a kit for
use in measuring the activity of a ABC transporter or a fragment
thereof in a biological sample in vitro or in vivo comprising (i) a
composition comprising a compound of formulae (I, Ic, Id, II, Ila,
IIb, IIc, and IId) or any of the above embodiments; and (ii)
instructions for a.) contacting the composition with the biological
sample and b.) measuring activity of said ABC transporter or a
fragment thereof. In one embodiment, the kit further comprises
instructions for a.) contacting an additional composition with the
biological sample; b.) measuring the activity of said ABC
transporter or a fragment thereof in the presence of said
additional compound, and c.) comparing the activity of the ABC
transporter in the presence of the additional compound with the
density of the ABC transporter in the presence of a composition of
formulae (I, Ic, Id, II, IIa, IIb, IIc, and IId). In preferred
embodiments, the kit is used to measure the density of CFTR.
[0225] In order that the invention described herein may be more
fully understood, the following examples are set forth. It should
be understood that these examples are for illustrative purposes
only and are not to be construed as limiting this invention in any
manner.
VI. Preparations and Examples
General Procedure I
Carboxylic Acid Building Block
##STR00398##
[0227] Benzyltriethylammonium chloride (0.025 equivalents) and the
appropriate dihalo compound (2.5 equivalents) were added to a
substituted phenyl acetonitrile. The mixture was heated at
70.degree. C. and then 50% sodium hydroxide (10 equivalents) was
slowly added to the mixture. The reaction was stirred at 70.degree.
C. for 12-24 hours to ensure complete formation of the cycloalkyl
moiety and then heated at 130.degree. C. for 24-48 hours to ensure
complete conversion from the nitrile to the carboxylic acid. The
dark brown/black reaction mixture was diluted with water and
extracted with dichloromethane three times to remove side products.
The basic aqueous solution was acidified with concentrated
hydrochloric acid to pH less than one and the precipitate which
began to form at pH 4 was filtered and washed with 1 M hydrochloric
acid two times. The solid material was dissolved in dichloromethane
and extracted two times with 1 M hydrochloric acid and one time
with a saturated aqueous solution of sodium chloride. The organic
solution was dried over sodium sulfate and evaporated to dryness to
give the cycloalkylcarboxylic acid. Yields and purities were
typically greater than 90%.
Example 1
1-Benzo[1,3]dioxol-5-yl-cyclopropanecarboxylic acid
##STR00399##
[0229] A mixture of 2-(benzo[d][1,3]dioxol-5-yl)acetonitrile (5.10
g 31.7 mmol), 1-bromo-2-chloro-ethane (9.00 mL 109 mmol), and
benzyltriethylammonium chloride (0.181 g, 0.795 mmol) was heated at
70.degree. C. and then 50% (wt/wt.) aqueous sodium hydroxide (26
mL) was slowly added to the mixture. The reaction was stirred at
70.degree. C. for 24 hours and then heated at 130.degree. C. for 48
hours. The dark brown reaction mixture was diluted with water (400
mL) and extracted once with an equal volume of ethyl acetate and
once with an equal volume of dichloromethane. The basic aqueous
solution was acidified with concentrated hydrochloric acid to pH
less than one and the precipitate filtered and washed with 1 M
hydrochloric acid. The solid material was dissolved in
dichloromethane (400 mL) and extracted twice with equal volumes of
I M hydrochloric acid and once with a saturated aqueous solution of
sodium chloride. The organic solution was dried over sodium sulfate
and evaporated to dryness to give a white to slightly off-white
solid (5.23 g, 80%) ESI-MS m/z calc. 206.1, found 207.1
(M+1).sup.+. Retention time 2.37 minutes. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 1.07-1.11 (m, 2H), 1.38-1.42 (m, 2H), 5.98
(s, 2H), 6.79 (m, 2H), 6.88 (m, 1H), 12.26 (s, 1H).
General Procedure II
Carboxylic Acid Building Block
##STR00400##
[0231] Sodium hydroxide (50% aqueous solution, 7.4 equivalents) was
slowly added to a mixture of the appropriate phenyl acetonitrile,
benzyltriethylammonium chloride (1.1 equivalents), and the
appropriate dihalo compound (2.3 equivalents) at 70.degree. C. The
mixture was stirred overnight at 70.degree. C. and the reaction
mixture was diluted with water (30 mL) and extracted with ethyl
acetate. The combined organic layers were dried over sodium sulfate
and evaporated to dryness to give the crude
cyclopropanecarbonitrile, which was used directly in the next
step.
[0232] The crude cyclopropanecarbonitrile was refluxed in 10%
aqueous sodium hydroxide (7.4 equivalents) for 2.5 hours. The
cooled reaction mixture was washed with ether (100 mL) and the
aqueous phase was acidified to pH 2 with 2M hydrochloric acid. The
precipitated solid was filtered to give the cyclopropanecarboxylic
acid as a white solid.
General Procedure III
Carboxylic Acid Building Block
##STR00401##
[0233] Example 2
1-(2,2-Difluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarboxylic
##STR00402##
[0234] 2,2-Difluoro-benzo[1,3]dioxole-5-carboxylic acid methyl
ester
[0235] A solution of 5-bromo-2,2-difluoro-benzo[1,3]dioxole (11.8
g, 50.0 mmol) and tetrakis(triphenylphosphine)palladium (0)
[Pd(PPh.sub.3).sub.4, 5.78 g, 5.00 mmol] in methanol (20 mL)
containing acetonitrile (30 mL) and triethylamine (10 mL) was
stirred under a carbon monoxide atmosphere (55 PSI) at 75.degree.
C. (oil bath temperature) for 15 hours. The cooled reaction mixture
was filtered and the filtrate was evaporated to dryness. The
residue was purified by silica gel column chromatography to give
crude 2,2-difluoro-benzo[1,3]dioxole-5-carboxylic acid methyl ester
(11.5 g), which was used directly in the next step.
##STR00403##
(2,2-Difluoro-benzo[1,3]dioxol-5-yl)-methanol
[0236] Crude 2,2-difluoro-benzo[1,3]dioxole-5-carboxylic acid
methyl ester (11.5 g) dissolved in 20 mL of anhydrous
tetrahydrofuran (THF) was slowly added to a suspension of lithium
aluminum hydride (4.10 g, 106 mmol) in anhydrous THF (100 mL) at
0.degree. C. The mixture was then warmed to room temperature. After
being stirred at room temperature for 1 hour, the reaction mixture
was cooled to 0.degree. C. and treated with water (4.1 g), followed
by sodium hydroxide (10% aqueous solution, 4.1 mL). The resulting
slurry was filtered and washed with THF. The combined filtrate was
evaporated to dryness and the residue was purified by silica gel
column chromatography to give
(2,2-difluoro-benzo[1,3]dioxol-5-yl)-methanol (7.2 g, 38 mmol, 76%
over two steps) as a colorless oil.
##STR00404##
5-Chloromethyl-2,2-difluoro-benzo[1,3]dioxole
[0237] Thionyl chloride (45 g, 38 mmol) was slowly added to a
solution of (2,2-difluoro-benzo[1,3]dioxol-5-yl)-methanol (7.2 g,
38 mmol) in dichloromethane (200 mL) at 0.degree. C. The resulting
mixture was stirred overnight at room temperature and then
evaporated to dryness. The residue was partitioned between an
aqueous solution of saturated sodium bicarbonate (100 mL) and
dichloromethane (100 mL). The separated aqueous layer was extracted
with dichloromethane (150 mL) and the organic layer was dried over
sodium sulfate, filtrated, and evaporated to dryness to give crude
5-chloromethyl-2,2-difluoro-benzo[1,3]dioxole (4.4 g) which was
used directly in the next step.
##STR00405##
(2,2-Difluoro-benzo[1,3]dioxol-5-yl)-acetonitrile
[0238] A mixture of crude
5-chloromethyl-2,2-difluoro-benzo[1,3]dioxole (4.4 g) and sodium
cyanide (1.36 g, 27.8 mmol) in dimethylsulfoxide (50 mL) was
stirred at room temperature overnight. The reaction mixture was
poured into ice and extracted with ethyl acetate (300 mL). The
organic layer was dried over sodium sulfate and evaporated to
dryness to give crude
(2,2-difluoro-benzo[1,3]dioxol-5-yl)-acetonitrile (3.3 g) which was
used directly in the next step.
##STR00406##
1-(2,2-Difluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarbonitrile
[0239] Sodium hydroxide (50% aqueous solution, 10 mL) was slowly
added to a mixture of crude
(2,2-difluoro-benzo[1,3]dioxol-5-yl)-acetonitrile,
benzyltriethylammonium chloride (3.00 g, 15.3 mmol), and
1-bromo-2-chloroethane (4.9 g, 38 mmol) at 70.degree. C.
[0240] The mixture was stirred overnight at 70.degree. C. before
the reaction mixture was diluted with water (30 mL) and extracted
with ethyl acetate. The combined organic layers were dried over
sodium sulfate and evaporated to dryness to give crude
1-(2,2-difluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarbonitrile,
which was used directly in the next step.
##STR00407##
1-(2,2-Difluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarboxylic
acid
[0241]
1-(2,2-Difluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarbonitrile
(crude from the last step) was refluxed in 10% aqueous sodium
hydroxide (50 mL) for 2.5 hours. The cooled reaction mixture was
washed with ether (100 mL) and the aqueous phase was acidified to
pH 2 with 2M hydrochloric acid. The precipitated solid was filtered
to give
1-(2,2-difluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarboxylic acid
as a white solid (0.15 g, 1.6% over four steps). ESI-MS m/z calc.
242.04, found 241.58 (M+1).sup.+; .sup.1H NMR (CDCl.sub.3) .delta.
7.14-7.04 (m, 2H), 6.98-6.96 (m, 1H), 1.74-1.64 (m, 2H), 1.26-1.08
(m, 2H).
Example 3
2-(2,2-Dimethylbenzo[d][1,3]dioxol-5-yl)acetonitrile
##STR00408##
[0242] (3,4-Dihydroxy-phenyl)-acetonitrile
[0243] To a solution of benzo[1,3]dioxol-5-yl-acetonitrile (0.50 g,
3.1 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added dropwise BBr.sub.3
(0.78 g, 3.1 mmol) at -78.degree. C. under N.sub.2. The mixture was
slowly warmed to room temperature and stirred overnight. H.sub.2O
(10 mL) was added to quench the reaction and the CH.sub.2Cl.sub.2
layer was separated. The aqueous phase was extracted with
CH.sub.2Cl.sub.2 (2.times.7 mL). The combined organics were washed
with brine, dried over Na.sub.2SO.sub.4 and purified by column
chromatography on silica gel (petroleum ether/ethyl acetate 5:1) to
give (3,4-dihydroxy-phenyl)-acetonitrile (0.25 g, 54%) as a white
solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 9.07 (s, 1H),
8.95 (s, 1H), 6.68-6.70 (m, 2H), 6.55 (dd, J=8.0, 2.0 Hz, 1H), 3.32
(s, 2H).
##STR00409##
2-(2,2-Dimethylbenzo[d][1,3]dioxol-5-yl)acetonitrile
[0244] To a solution of (3,4-dihydroxy-phenyl)-acetonitrile (0.20
g, 1.3 mmol) in toluene (4 mL) was added 2,2-dimethoxy-propane
(0.28 g, 2.6 mmol) and TsOH (0.010 g, 0.065 mmol). The mixture was
heated at reflux overnight. The reaction mixture was evaporated to
remove the solvent and the residue was dissolved in ethyl acetate.
The organic layer was washed with NaHCO.sub.3 solution, H.sub.2O,
brine, and dried over Na.sub.2SO.sub.4. The solvent was evaporated
under reduced pressure to give a residue, which was purified by
column chromatography on silica gel (petroleum ether/ethyl acetate
10:1) to give 2-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)acetonitrile
(40 mg, 20%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 6.68-6.71
(m, 3H), 3.64 (s, 2H), 1.67 (s, 6H).
Example 4
1-(3,4-Dihydroxy-phenyl)-cyclopropanecarboxylic acid
##STR00410##
[0245] 1-(3,4-Bis-benzyloxy-phenyl)-cyclopropanecarbonitrile
[0246] To a mixture of (n-C.sub.4H.sub.9).sub.4NBr (0.50 g, 1.5
mmol), toluene (7 mL) and (3,4-bis-benzyloxy-phenyl)-acetonitrile
(14 g, 42 mmol) in NaOH (50 g) and H.sub.2O (50 mL) was added
BrCH.sub.2CH.sub.2Cl (30 g, 0.21 mol). The reaction mixture was
stirred at 50.degree. C. for 5 h before being cooled to room
temperature. Toluene (30 mL) was added and the organic layer was
separated and washed with H.sub.2O, brine, dried over anhydrous
MgSO.sub.4, and concentrated. The residue was purified by column on
silica gel (petroleum ether/ethyl acetate 10:1) to give
1-(3,4-bis-benzyloxy-phenyl)-cyclopropanecarbonitrile (10 g, 66%).
.sup.1H NMR (DMSO 300 MHz) .delta. 7.46-7.30 (m, 10H), 7.03 (d,
J=8.4 Hz, 1H), 6.94 (d, J=2.4 Hz, 1H), 6.89 (dd, J=2.4, 8.4 Hz,
1H), 5.12 (d, J=7.5 Hz, 4H), 1.66-1.62 (m, 2H), 1.42-1.37 (m,
2H).
##STR00411##
1-(3,4-Dihydroxy-phenyl)-cyclopropanecarbonitrile
[0247] To a solution of
1-(3,4-bis-benzyloxy-phenyl)-cyclopropanecarbonitrile (10 g, 28
mmol) in MeOH (50 mL) was added Pd/C (0.5 g) under nitrogen
atmosphere. The mixture was stirred under hydrogen atmosphere (1
atm) at room temperature for 4 h. The catalyst was filtered off
through a celite pad and the filtrate was evaporated under vacuum
to give 1-(3,4-dihydroxy-phenyl)-cyclopropanecarbonitrile (4.5 g,
92%). .sup.1H NMR (DMSO 400 MHz) .delta. 9.06 (br s, 2H), 6.67-6.71
(m, 2H), 6.54 (dd, J=2.4, 8.4 Hz, 1H), 1.60-1.57 (m, 2H), 1.30-1.27
(m, 2H).
##STR00412##
1-(3,4-Dihydroxy-phenyl)-cyclopropanecarboxylic acid
[0248] To a solution of NaOH (20 g, 0.50 mol) in H.sub.2O (20 mL)
was added 1-(3,4-dihydroxy-phenyl)-cyclopropanecarbonitrile (4.4 g,
25 mmol). The mixture was heated at reflux for 3 h before being
cooled to room temperature. The mixture was neutralized with HCl
(0.5 N) to pH 3-4 and extracted with ethyl acetate (20 mL.times.3).
The combined organic layers were washed with water, brine, dried
over anhydrous MgSO.sub.4, and concentrated under vacuum to obtain
1-(3,4-dihydroxy-phenyl)-cyclopropanecarboxylic acid (4.5 g crude).
From 900 mg crude, 500 mg pure
1-(3,4-dihydroxy-phenyl)-cyclopropanecarboxylic acid was obtained
by preparatory HPLC. .sup.1H NMR (DMSO, 300 MHz) .delta. 12.09 (br
s, 1H), 8.75 (br s, 2H), 6.50-6.67 (m, 3H), 1.35-1.31 (m, 2H),
1.01-0.97 (m, 2H).
Example 5
1-(2-Oxo-2,3-dihydrobenzo[d]oxazol-5-yl)cyclopropane-carboxylic
acid
##STR00413##
[0249] 1-(4-Methoxy-phenyl)-cyclopropanecarboxylic acid methyl
ester
[0250] To a solution of 1-(4-methoxy-phenyl)-cyclopropanecarboxylic
acid (50 g, 0.26 mol) in MeOH (500 mL) was added toluene-4-sulfonic
acid monohydrate (2.5 g, 13 mmol) at room temperature. The reaction
mixture was heated at reflux for 20 hours. MeOH was removed by
evaporation under vacuum and EtOAc (200 mL) was added. The organic
layer was washed with sat. aq. NaHCO.sub.3 (100 mL) and brine,
dried over anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum
to give 1-(4-methoxy-phenyl)-cyclopropanecarboxylic acid methyl
ester (53 g, 99%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
7.25-7.27 (m, 2H), 6.85 (d, J=8.8 Hz, 2H), 3.80 (s, 3H), 3.62 (s,
3H), 1.58 (q, J=3.6 Hz, 2H), 1.15 (q, J=3.6 Hz, 2H).
##STR00414##
1-(4-Methoxy-3-nitro-phenyl)-cyclopropanecarboxylic acid methyl
ester
[0251] To a solution of 1-(4-methoxy-phenyl)-cyclopropanecarboxylic
acid methyl ester (30.0 g, 146 mmol) in Ac.sub.2O (300 mL) was
added a solution of HNO.sub.3 (14.1 g, 146 mmol, 65%) in AcOH (75
mL) at 0.degree. C. The reaction mixture was stirred at
0.about.5.degree. C. for 3 h before aq. HCl (20%) was added
dropwise at 0.degree. C. The resulting mixture was extracted with
EtOAc (200 mL.times.3). The organic layer was washed with sat. aq.
NaHCO.sub.3 then brine, dried over anhydrous Na.sub.2SO.sub.4 and
evaporated under vacuum to give
1-(4-methoxy-3-nitro-phenyl)-cyclopropanecarboxylic acid methyl
ester (36.0 g, 98%), which was directly used in the next step.
.sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 7.84 (d, J=2.1 Hz, 1H),
7.54 (dd, J=2.1, 8.7 Hz, 1H), 7.05 (d, J=8.7 Hz, 1H), 3.97 (s, 3H),
3.65 (s, 3H), 1.68-1.64 (m, 2H), 1.22-1.18 (m, 2H).
##STR00415##
1-(4-Hydroxy-3-nitro-phenyl)-cyclopropanecarboxylic acid methyl
ester
[0252] To a solution of
1-(4-methoxy-3-nitro-phenyl)-cyclopropane-carboxylic acid methyl
ester (10.0 g, 39.8 mmol) in CH.sub.2Cl.sub.2 (100 mL) was added
BBr.sub.3 (12.0 g, 47.8 mmol) at -70.degree. C. The mixture was
stirred at -70.degree. C. for 1 hour, then allowed to warm to
-30.degree. C. and stirred at this temperature for 3 hours. Water
(50 mL) was added dropwise at -20.degree. C., and the resulting
mixture was allowed to warm room temperature before it was
extracted with EtOAc (200 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4 and evaporated under
vacuum to give the crude product, which was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate 15:1)
to afford 1-(4-hydroxy-3-nitro-phenyl)-cyclopropanecarboxylic acid
methyl ester (8.3 g, 78%). .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 10.5 (s, 1H), 8.05 (d, J=2.4 Hz, 1H), 7.59 (dd, J=2.0, 8.8
Hz, 1H), 7.11 (d, J=8.4 Hz, 1H), 3.64 (s, 3H), 1.68-1.64 (m, 2H),
1.20-1.15 (m, 2H).
##STR00416##
1-(3-Amino-4-hydroxy-phenyl)-cyclopropanecarboxylic acid methyl
ester
[0253] To a solution of
1-(4-hydroxy-3-nitro-phenyl)-cyclopropanecarboxylic acid methyl
ester (8.3 g, 35 mmol) in MeOH (100 mL) was added Raney Nickel (0.8
g) under nitrogen atmosphere. The mixture was stirred under
hydrogen atmosphere (1 atm) at 35.degree. C. for 8 hours. The
catalyst was filtered off through a Celite pad and the filtrate was
evaporated under vacuum to give crude product, which was purified
by column chromatography on silica gel (petroleum ether/ethyl
acetate 1:1) to give
1-(3-amino-4-hydroxy-phenyl)-cyclopropanecarboxylic acid methyl
ester (5.3 g, 74%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 6.77
(s, 1H), 6.64 (d, J=2.0 Hz, 2H), 3.64 (s, 3H), 1.55-1.52 (m, 2H),
1.15-1.12 (m, 2H).
##STR00417##
1-(2-Oxo-2,3-dihydro-benzooxazol-5-yl)-cyclopropanecarboxylic acid
methyl ester
[0254] To a solution of
1-(3-amino-4-hydroxy-phenyl)-cyclopropanecarboxylic acid methyl
ester (2.0 g, 9.6 mmol) in THF (40 mL) was added triphosgene (4.2
g, 14 mmol) at room temperature. The mixture was stirred for 20
minutes at this temperature before water (20 mL) was added dropwise
at 0.degree. C. The resulting mixture was extracted with EtOAc (100
mL.times.3). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under vacuum to give
1-(2-oxo-2,3-dihydro-benzooxazol-5-yl)-cyclopropanecarboxylic acid
methyl ester (2.0 g, 91%), which was directly used in the next
step. .sup.1H NMR (CDC.sub.3, 300 MHz) .delta. 8.66 (s, 1H),
7.13-7.12 (m, 2H), 7.07 (s, 1H), 3.66 (s, 3H), 1.68-1.65 (m, 2H),
1.24-1.20 (m, 2H).
##STR00418##
1-(2-Oxo-2,3-dihydrobenzo[d]oxazol-5-yl)cyclopropanecarboxylic
acid
[0255] To a solution of
1-(2-oxo-2,3-dihydro-benzooxazol-5-yl)-cyclopropanecarboxylic acid
methyl ester (1.9 g, 8.1 mmol) in MeOH (20 mL) and water (2 mL) was
added LiOH.H.sub.2O (1.7 g, 41 mmol) in portions at room
temperature. The reaction mixture was stirred for 20 hours at
50.degree. C. MeOH was removed by evaporation under vacuum before
water (100 mL) and EtOAc (50 mL) were added. The aqueous layer was
separated, acidified with HCl (3 mol/L) and extracted with EtOAc
(100 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum to give
1-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)cyclopropanecarboxylic acid
(1.5 g, 84%). .sup.1H NMR (DMSO, 400 MHz) .delta. 12.32 (brs, 1H),
11.59 (brs, 1H), 7.16 (d, J=8.4 Hz, 1H), 7.00 (d, J=8.0 Hz, 1H),
1.44-1.41 (m, 2H), 1.13-1.10 (m, 2H). MS (ESI) m/e (M+H.sup.4)
218.1.
Example 6
1-(6-Fluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarboxylic acid
##STR00419##
[0256] 2-Fluoro-4,5-dihydroxy-benzaldehyde
[0257] To a stirred suspension of
2-fluoro-4,5-dimethoxy-benzaldehyde (3.00 g, 16.3 mmol) in
dichloromethane (100 mL) was added BBrs (12.2 mL, 130 mmol)
dropwise at -78.degree. C. under nitrogen atmosphere. After
addition, the mixture was warmed to -30.degree. C. and stirred at
this temperature for 5 h. The reaction mixture was poured into ice
water and the precipitated solid was collected by filtration and
washed with dichloromethane to afford
2-fluoro-4,5-dihydroxy-benzaldehyde (8.0 g), which was used
directly in the next step.
##STR00420##
6-Fluoro-benzo[1,3]dioxole-5-carbaldehyde
[0258] To a stirred solution of 2-fluoro-4,5-dihydroxy-benzaldehyde
(8.0 g) and BrClCH.sub.2 (24.8 g, 190 mmol) in dry DMF (50 mL) was
added Cs.sub.2CO.sub.3 (62.0 g, 190 mmol) in portions. The
resulting mixture was stirred at 60.degree. C. overnight and then
poured into water. The mixture was extracted with EtOAc (200 mL x
3). The combined organic layers were washed with brine (200 mL),
dried over Na.sub.2SO.sub.4, and evaporated in vacuo to give crude
product, which was purified by column chromatography on silica gel
(5-20% ethyl acetate/petroleum ether) to afford
6-fluoro-benzo[1,3]dioxole-5-carbaldehyde (700 mg, two steps yield:
24%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 10.19 (s, 1H), 7.23
(d, J=5.6, 1H), 6.63 (d, J=9.6, 1H), 6.08 (s, 2H).
##STR00421##
(6-Fluoro-benzo[1,3]dioxol-5-yl)-methanol
[0259] To a stirred solution of
6-fluoro-benzo[1,3]dioxole-5-carbaldehyde (700 mg, 4.2 mmol) in
MeOH (50 mL) was added NaBH.sub.4 (320 mg, 8.4 mmol) in portions at
0.degree. C. The mixture was stirred at this temperature for 30 min
and was then concentrated in vacuo to give a residue. The residue
was dissolved in EtOAc and the organic layer was washed with water,
dried over Na.sub.2SO.sub.4, and concentrated in vacuo to afford
(6-fluoro-benzo[1,3]dioxol-5-yl)-methanol (650 mg, 92%), which was
directly used in the next step.
##STR00422##
5-Chloromethyl-6-fluoro-benzo[1,3]dioxole
[0260] (6-Fluoro-benzo[1,3]dioxol-5-yl)-methanol (650 mg, 3.8 mmol)
was added to SOCl.sub.2 (20 mL) in portions at 0.degree. C. The
mixture was warmed to room temperature for 1 h and then heated at
reflux for 1 h. The excess SOCl.sub.2 was evaporated under reduced
pressure to give the crude product, which was basified with sat.
NaHCO.sub.3 solution to pH.about.7. The aqueous phase was extracted
with EtOAc (50 mL.times.3). The combined organic layers were dried
over Na.sub.2SO.sub.4 and evaporated under reduced pressure to give
5-chloromethyl-6-fluoro-benzo[1,3]dioxole (640 mg, 90%), which was
directly used in the next step.
##STR00423##
(6-Fluoro-benzo[1,3]dioxol-5-yl)-acetonitrile
[0261] A mixture of 5-chloromethyl-6-fluoro-benzo[1,3]dioxole (640
mg, 3.4 mmol) and NaCN (340 mg, 6.8 mmol) in DMSO (20 mL) was
stirred at 30.degree. C. for 1 h and then poured into water. The
mixture was extracted with EtOAc (50 mL.times.3). The combined
organic layers were washed with water (50 mL) and brine (50 mL),
dried over Na.sub.2SO.sub.4, and evaporated under reduced pressure
to give the crude product, which was purified by column
chromatography on silica gel (5-10% ethyl acetate/petroleum ether)
to afford (6-fluoro-benzo[1,3]dioxol-5-yl)-acetonitrile (530 mg,
70%). .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta. 6.82 (d, J=4.8,
1H), 6.62 (d, J=5.4, 1H), 5.99 (s, 2H), 3.65 (s, 2H).
##STR00424##
1-(6-Fluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarbonitrile
[0262] A flask was charged with water (10 mL), followed by a rapid
addition of NaOH (10 g, 0.25 mol) in three portions over a 5 min
period. The mixture was allowed to cool to room temperature.
Subsequently, the flask was charged with toluene (6 mL),
tetrabutyl-ammonium bromide (50 mg, 0.12 mmol),
(6-fluoro-benzo[1,3]dioxol-5-yl)-acetonitrile (600 mg, 3.4 mmol)
and 1-bromo-2-chloroethane (1.7 g, 12 mmol). The mixture stirred
vigorously at 50.degree. C. overnight. The cooled flask was charged
with additional toluene (20 mL). The organic layer was separated
and washed with water (30 mL) and brine (30 mL). The organic layer
was removed in vacuo to give the crude product, which was purified
by column chromatography on silica gel (5-10% ethyl
acetate/petroleum ether) to give
1-(6-fluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarbonitrile (400
mg, 60%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 6.73 (d, J=3.0
Hz, 1H), 6.61 (d, J=9.3 Hz, 1H), 5.98 (s, 2H), 1.67-1.62 (m, 2H),
1.31-1.27 (m, 2H).
##STR00425##
1-(6-Fluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarboxylic acid
[0263] A mixture of
L-(6-fluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarbonitrile (400
mg, 0.196 mmol) and 10% NaOH (10 mL) was stirred at 100.degree. C.
overnight. After the reaction was cooled, 5% HCl was added until
the pH<5 and then EtOAc (30 mL) was added to the reaction
mixture. The layers were separated and combined organic layers were
evaporated in vacuo to afford
1-(6-fluoro-benzo[1,3]dioxol-5-yl)-cyclopropanecarboxylic acid (330
mg, 76%). .sup.1H NMR (400 MHz, DMSO) .delta. 12.2 (s, 1H),
6.87-6.85 (m, 2H), 6.00 (s, 1H), 1.42-1.40 (m, 2H), 1.14-1.07 (m,
2H).
Example 7
1-(Benzofuran-5-yl)cyclopropanecarboxylic acid
##STR00426##
[0264] 1-[4-(2,2-Diethoxy-ethoxy)-phenyl]-cyclopropanecarboxylic
acid
[0265] To a stirred solution of
1-(4-hydroxy-phenyl)-cyclopropanecarboxylic acid methyl ester (15.0
g, 84.3 mmol) in DMF (50 mL) was added sodium hydride (6.7 g, 170
mmol, 60% in mineral oil) at 0.degree. C. After hydrogen evolution
ceased, 2-bromo-1,1-diethoxy-ethane (16.5 g, 84.3 mmol) was added
dropwise to the reaction mixture. The reaction was stirred at
160.degree. C. for 15 hours. The reaction mixture was poured onto
ice (100 g) and was extracted with CH.sub.2Cl.sub.2. The combined
organics were dried over Na.sub.2SO.sub.4. The solvent was
evaporated under vacuum to give
1-[4-(2,2-diethoxy-ethoxy)-phenyl]-cyclopropanecarboxylic acid (10
g), which was used directly in the next step without
purification.
##STR00427##
1-Benzofuran-S-yl-cyclopropanecarboxylic acid
[0266] To a suspension of
1-[4-(2,2-diethoxy-ethoxy)-phenyl]-cyclopropenecarboxylic acid (20
g, .about.65 mmol) in xylene (100 mL) was added PPA (22.2 g, 64.9
mmol) at room temperature. The mixture was heated at reflux
(140.degree. C.) for 1 hour before it was cooled to room
temperature and decanted from the PPA. The solvent was evaporated
under vacuum to obtain the crude product, which was purified by
preparative HPLC to provide
1-(benzofuran-5-yl)cyclopropanecarboxylic acid (1.5 g, 5%). .sup.1H
NMR (400 MHz, DMSO-ds) .delta. 12.25 (br s, 1H), 7.95 (d, J=2.8 Hz,
1H), 7.56 (d, J=2.0 Hz, 1H), 7.47 (d, J=11.6 Hz, 1H), 7.25 (dd,
J=2.4, 11.2 Hz, 1H), 6.89 (d, J=1.6 Hz, 1H), 1.47-1.44 (m, 2H),
1.17-1.14 (m, 2H).
Example 8
1-(2,3-Dihydrobenzofuran-6-yl)cyclopropanecarboxylic acid
##STR00428##
[0268] To a solution of 1-(benzofuran-6-yl)cyclopropanecarboxylic
acid (370 mg, 1.8 mmol) in MeOH (50 mL) was added PtO.sub.2 (75 mg,
20%) at room temperature. The reaction mixture was stirred under
hydrogen atmosphere (1 atm) at 20.degree. C. for 3 d. The reaction
mixture was filtered and the solvent was evaporated in vacuo to
afford the crude product, which was purified by prepared HPLC to
give 1-(2,3-dihydrobenzofuran-6-yl)cyclopropanecarboxylic acid (155
mg, 42%). .sup.1H NMR (300 MHz, MeOD) .delta. 7.13 (d; J=7.5 Hz,
1H), 6.83 (d, J=7.8 Hz, 1H), 6.74 (s, 1H), 4.55 (t, J=8.7 Hz, 2H),
3.18 (t, J=8.7 Hz, 2H), 1.56-1.53 (m, 2H), 1.19-1.15 (m, 2H).
Example 9
1-(3,3-Dimethyl-2,3-dihydrobenzofuran-5-yl)cyclopropanecarboxylic
acid
##STR00429##
[0270] To a solution of methyl
1-(4-methoxyphenyl)cyclopropanecarboxylate (10.0 g, 48.5 mmol) in
dichloromethane (80 mL) was added EtSH (16 mL) under ice-water
bath. The mixture was stirred at 0.degree. C. for 20 min before
AlCl.sub.3 (19.5 g, 0.15 mmol) was added slowly at 0.degree. C. The
mixture was stirred at 0.degree. C. for 30 min. The reaction
mixture was poured into ice-water, the organic layer was separated,
and the aqueous phase was extracted with dichloromethane (50
mL.times.3). The combined organic layers were washed with H.sub.2O,
brine, dried over Na.sub.2SO.sub.4 and evaporated under vacuum to
give 1-(4-hydroxy-phenyl)-cyclopropanecarboxylic acid methyl ester
(8.9 g, 95%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.20-7.17
(m, 2H), 6.75-6.72 (m, 2H), 5.56 (s, 1H), 3.63 (s, 3H), 1.60-1.57
(m, 2H), 1.17-1.15 (m, 2H).
##STR00430##
1-(4-Hydroxy-3,5-diiodo-phenyl)-cyclopropanecarboxylic acid methyl
ester
[0271] To a solution of 1-(4-hydroxy-phenyl)-cyclopropanecarboxylic
acid methyl ester (8.9 g, 46 mmol) in CH.sub.3CN (80 mL) was added
NIS (15.6 g, 69 mmol). The mixture was stirred at room temperature
for 1 hour. The reaction mixture was concentrated and the residue
was purified by column chromatography on silica gel (petroleum
ether/ethyl acetate 10:1) to give
1-(4-hydroxy-3,5-diiodo-phenyl)-cyclopropanecarboxylic acid methyl
ester (3.5 g, 18%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.65
(s, 2H), 5.71 (s, 1H), 3.63 (s, 3H), 1.59-1.56 (m, 2H), 1.15-1.12
(m, 2H).
##STR00431##
1-(3,5-Diiodo-4-(2-methyl-allyloxy)-phenyl]-cyclopropanecarboxylic
acid methyl ester
[0272] A mixture of
1-(4-hydroxy-3,5-diiodo-phenyl)-cyclopropanecarboxylic acid methyl
ester (3.2 g, 7.2 mmol), 3-chloro-2-methyl-propene (1.0 g, 11
mmol), K.sub.2CO.sub.3 (1.2 g, 8.6 mmol), NaI (0.1 g, 0.7 mmol) in
acetone (20 mL) was stirred at 20.degree. C. overnight. The solid
was filtered off and the filtrate was concentrated under vacuum to
give
1-[3,5-diiodo-4-(2-methyl-allyloxy)-phenyl]-cyclopropane-carboxylic
acid methyl ester (3.5 g, 97%). .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 7.75 (s, 2H), 5.26 (s, 1H), 5.06 (s, 1H), 4.38 (s, 2H),
3.65 (s, 3H), 1.98 (s, 3H), 1.62-1.58 (m, 2H), 1.18-1.15 (m,
2H).
##STR00432##
1-(3,3-Dimethyl-2,3-dihydro-benzofuran-5-yl)-cyclopropanecarboxylic
acid methyl ester
[0273] To a solution of
1-[3,5-diiodo-4-(2-methyl-allyloxy)-phenyl]-cyclopropane-carboxylic
acid methyl ester (3.5 g, 7.0 mmol) in toluene (15 mL) was added
Bu.sub.3SnH (2.4 g, 8.4 mmol) and AIBN (0.1 g, 0.7 mmol). The
mixture was heated at reflux overnight. The reaction mixture was
concentrated under vacuum and the residue was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate 20:1)
to give
1-(3,3-dimethyl-2,3-dihydro-benzofuran-5-yl)-cyclopropanecarboxylic
acid methyl ester (1.05 g, 62%). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.10-7.07 (m, 2H), 6.71 (d, J=8 Hz, 1H), 4.23 (s, 2H), 3:62
(s, 3H), 1.58-1.54 (m, 2H), 1.34 (s, 6H), 1.17-1.12 (m, 2H).
##STR00433##
1-(3,3-Dimethyl-2,3-dihydrobenzofuran-5-yl)cyclopropanecarboxylic
acid
[0274] To a solution of
1-(3,3-dimethyl-2,3-dihydro-benzofuran-5-yl)-cyclopropanecarboxylic
acid methyl ester (1.0 g, 4.0 mmol) in MeOH (10 mL) was added LiOH
(0.40 g, 9.5 mmol). The mixture was stirred at 40.degree. C.
overnight. HCl (10%) was added slowly to adjust the pH to 5. The
resulting mixture was extracted with ethyl acetate (10 mL x 3). The
extracts were washed with brine and dried over Na.sub.2SO.sub.4.
The solvent was removed under vaccum and the crude product was
purified by preparative HPLC to give
1-(3,3-dimethyl-2,3-dihydrobenzofuran-5-yl)cyclopropanecarboxylic
acid (0.37 g, 41%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.11-7.07 (m, 2H), 6.71 (d, J=8 Hz, 1H), 4.23 (s, 2H), 1.66-1.63
(m, 2H), 1.32 (s, 6H), 1.26-1.23 (m, 2H).
Example 10
2-(7-Methoxybenzo[d][1,3]dioxol-5-yl)acetonitrile
##STR00434##
[0275] 3,4-Dihydroxy-5-methoxybenzoate
[0276] To a solution of 3,4,5-trihydroxy-benzoic acid methyl ester
(50 g, 0.27 mol) and Na.sub.2B.sub.4O.sub.7 (50 g) in water (1000
mL) was added Me.sub.2SO.sub.4 (120 mL) and aqueous NaOH solution
(25%, 200 mL) successively at room temperature. The mixture was
stirred at room temperature for 6 h before it was cooled to
0.degree. C. The mixture was acidified to pH.about.2 by adding
cone. H.sub.2SO.sub.4 and then filtered. The filtrate was extracted
with EtOAc (500 mL.times.3). The combined organic layers were dried
over anhydrous Na.sub.2SO.sub.4 and evaporated under reduced
pressure to give methyl 3,4-dihydroxy-5-methoxybenzoate (15.3 g
47%), which was used in the next step without further
purification.
##STR00435##
Methyl 7-methoxybenzo[d][1,3]dioxole-5-carboxylate
[0277] To a solution of methyl 3,4-dihydroxy-5-methoxybenzoate
(15.3 g, 0.0780 mol) in acetone (500 mL) was added CH.sub.2BrCl
(34.4 g, 0.270 mol) and K.sub.2CO.sub.3 (75.0 g, 0.540 mol) at
80.degree. C. The resulting mixture was heated at reflux for 4 h.
The mixture was cooled to room temperature and solid
K.sub.2CO.sub.3 was filtered off. The filtrate was concentrated
under reduced pressure, and the residue was dissolved in EtOAc (100
mL). The organic layer was washed with water, dried over anhydrous
Na.sub.2SO.sub.4, and evaporated under reduced pressure to give the
crude product, which was purified by column chromatography on
silica gel (petroleum ether/ethyl acetate=10:1) to afford methyl
7-methoxybenzo[d][1,3]dioxole-5-carboxylate (12.6 g, 80%). .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.32 (s, 1H), 7.21 (s, 1H), 6.05
(s, 2H), 3.93 (s, 3H), 3.88 (s, 3H)
##STR00436##
(7-Methoxybenzo[d[[1,3]dioxol-5-yl)methanol
[0278] To a solution of methyl
7-methoxybenzo[d][1,3]dioxole-5-carboxylate (14 g, 0.040 mol) in
THF (100 mL) was added LiAlH.sub.4 (3.1 g, 0.080 mol) in portions
at room temperature. The mixture was stirred for 3 h at room
temperature. The reaction mixture was cooled to 0.degree. C. and
treated with water (3.1 g) and NaOH (10%, 3.1 mL) successively. The
slurry was filtered off and washed with THF. The combined filtrates
were evaporated under reduced pressure to give
(7-methoxy-benzo[d][1,3]dioxol-5-yl)methanol (7.2 g, 52%). .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 6.55 (s, 1H), 6.54 (s, 1H), 5.96
(s, 2H), 4.57 (s, 2H), 3.90 (s, 3H).
##STR00437##
6-(Chloromethyl)-4-methoxybenzo[d][1,3]dioxole
[0279] To a solution of SOCl.sub.2 (150 mL) was added
(7-methoxybenzo[d][(1,3]dioxol-5-yl)methanol (9.0 g, 54 mmol) in
portions at 0.degree. C. The mixture was stirred for 0.5 h. The
excess SOCl.sub.2 was evaporated under reduced pressure to give the
crude product, which was basified with sat. aq. NaHCO.sub.3 to
pH.about.7. The aqueous phase was extracted with EtOAc (100
mL.times.3). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4 and evaporated to give
6-(chloromethyl)-4-methoxybenzo[d][1,3]dioxole (10 g 94%), which
was used in the next step without further purification. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 6.58 (s, 1H), 6.57 (s, 1H), 5.98 (s,
2H), 4.51 (s, 2H), 3.90 (s 3H).
##STR00438##
2-(7-Methoxybenzo[d][1,3]dioxol-5-yl)acetonitrile
[0280] To a solution of
6-(chloromethyl)-4-methoxybenzo[d][1,3]dioxole (10 g, 40 mmol) in
DMSO (100 mL) was added NaCN (2.4 g, 50 mmol) at room temperature.
The mixture was stirred for 3 h and poured into water (500 mL). The
aqueous phase was extracted with EtOAc (100 mL.times.3). The
combined organic layers were dried over anhydrous Na.sub.2SO.sub.4
and evaporated to give the crude product, which was washed with
ether to afford 2-(7-methoxybenzo[d][1,3]dioxol-5-yl)acetonitrile
(4.6 g 45%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 6.49 (s,
2H), 5.98 (s, 2H), 3.91 (s, 3H), 3.65 (s, 2H). .sup.13C NMR (400
MHz, CDCl.sub.3) .delta. 148.9, 143.4, 134.6, 123.4, 117.3, 107.2,
101.8, 101.3, 56.3, 23.1.
Example 11
2-(3-(Benzyloxy)-4-methoxyphenyl)acetonitrile
##STR00439##
[0282] To a suspension of t-BuOK (20.2 g, 0.165 mol) in THF (250
mL) was added a solution of TosMIC (16.1 g, 82.6 mmol) in THF (100
mL) at -78.degree. C. The mixture was stirred for 15 minutes,
treated with a solution of 3-benzyloxy-4-methoxy-benzaldehyde (10.0
g, 51.9 mmol) in THF (50 mL) dropwise, and continued to stir for
1.5 hours at -78.degree. C. To the cooled reaction mixture was
added methanol (50 mL). The mixture was heated at reflux for 30
minutes. Solvent was removed to give a crude product, which was
dissolved in water (300 mL). The aqueous phase was extracted with
EtOAc (100 mL.times.3). The combined organic layers were dried and
evaporated under reduced pressure to give crude product, which was
purified by column chromatography (petroleum ether/ethyl acetate
10:1) to afford 2-(3-(benzyloxy)-4-methoxyphenyl)-acetonitrile (5.0
g, 48%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.48-7.33 (m,
5H), 6.89-6.86 (m, 3H), 5.17 (s, 2H), 3.90 (s, 3H), 3.66 (s, 2H).
.sup.13C NMR (75 MHz, CDCl.sub.3) .delta. 149.6, 148.6, 136.8,
128.8, 128.8, 128.2, 127.5, 127.5, 122.1, 120.9, 118.2, 113.8,
112.2, 71.2. 56.2, 23.3.
Example 12
2-(3-(Benzyloxy)-4-chlorophenyl)acetonitrile
##STR00440##
[0283] (4-Chloro-3-hydroxy-phenyl)acetonitrile
[0284] BBr.sub.3 (17 g, 66 mmol) was slowly added to a solution of
2-(4-chloro-3-methoxyphenyl)acetonitrile (12 g, 66 mmol) in
dichloromethane (120 mL) at -78.degree. C. under N.sub.2. The
reaction temperature was slowly increased to room temperature. The
reaction mixture was stirred overnight and then poured into ice and
water. The organic layer was separated, and the aqueous layer was
extracted with dichloromethane (40 mL.times.3). The combined
organic layers were washed with water, brine, dried over
Na.sub.2SO.sub.4, and concentrated under vacuum to give
(4-chloro-3-hydroxy-phenyl)-acetonitrile (9.3 g, 85%). .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 7.34 (d, J=8.4 Hz, 1H), 7.02 (d,
J=2.1 Hz, 1H), 6.87 (dd, J=2.1, 8.4 Hz, 1H), 5.15 (brs, 1H), 3.72
(s, 2H).
##STR00441##
2-(3-(Benzyloxy)-4-chlorophenyl)acetonitrile
[0285] To a solution of (4-chloro-3-hydroxy-phenyl)acetonitrile
(6.2 g, 37 mmol) in CH.sub.3CN (80 mL) was added K.sub.2CO.sub.3
(10 g, 74 mmol) and BnBr (7.6 g, 44 mmol). The mixture was stirred
at room temperature overnight. The solids were filtered off and the
filtrate was evaporated under vacuum. The residue was purified by
column chromatography on silica gel (petroleum ether/ethyl acetate
50:1) to give 2-(3-(benzyloxy)-4-chlorophenyl)-acetonitrile (5.6 g,
60%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.48-7.32 (m, 6H),
6.94 (d, J=2 Hz, 2H), 6.86 (dd, J=2.0, 8.4 Hz, 1H), 5.18 (s, 2H),
3.71 (s, 2H).
Example 13
2-(3-(Benzyloxy)-4-methoxyphenyl)acetonitrile
##STR00442##
[0287] To a suspension of t-BuOK (20.2 g, 0.165 mol) in THF (250
mL) was added a solution of TosMIC (16.1 g, 82.6 mmol) in THF (100
mL) at -78.degree. C. The mixture was stirred for 15 minutes,
treated with a solution of 3-benzyloxy-4-methoxy-benzaldehyde (10.0
g, 51.9 mmol) in THF (50 mL) dropwise, and continued to stir for
1.5 hours at -78.degree. C. To the cooled reaction mixture was
added methanol (50 mL). The mixture was heated at reflux for 30
minutes. Solvent of the reaction mixture was removed to give a
crude product, which was dissolved in water (300 mL). The aqueous
phase was extracted with EtOAc (100 mL.times.3). The combined
organic layers were dried and evaporated under reduced pressure to
give crude product, which was purified by column chromatography
(petroleum ether/ethyl acetate 10:1) to afford
2-(3-(benzyloxy)-4-methoxyphenyl)acetonitril (5.0 g, 48%). .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 7.48-7.33 (m, 5H), 6.89-6.86 (m,
3H), 5.17 (s, 2H), 3.90 (s, 3H), 3.66 (s, 2H). .sup.13C NMR (75
MHz, CDCl.sub.3) .delta. 149.6, 148.6, 136.8, 128.8, 128.8, 128.2,
127.5, 127.5, 122.1, 120.9, 118.2, 113.8, 112.2, 71.2, 56.2,
23.3.
Example 14
2-(3-Chloro-4-methoxyphenyl)acetonitrile
##STR00443##
[0289] To a suspension of t-BuOK (4.8 g, 40 mmol) in THF (30 mL)
was added a solution of TosMIC (3.9 g, 20 mmol) in THF (10 mL) at
-78.degree. C. The mixture was stirred for 10 minutes, treated with
a solution of 3-chloro-4-methoxy-benzaldehyde (1.7 g, 10 mmol) in
THF (10 mL) dropwise, and continued to stir for 1.5 hours at
-78.degree. C. To the cooled reaction mixture was added methanol
(10 mL). The mixture was heated at reflux for 30 minutes. Solvent
of the reaction mixture was removed to give a crude product, which
was dissolved in water (20 mL). The aqueous phase was extracted
with EtOAc (20 mL.times.3). The combined organic layers were dried
and evaporated under reduced pressure to give crude product, which
was purified by column chromatography (petroleum ether/ethyl
acetate 10:1) to afford 2-(3-chloro-4-methoxyphenyl)acetonitrile
(1.5 g, 83%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.33 (d,
J=2.4 Hz, 1H), 7.20 (dd, J=2.4, 8.4 Hz, 1H), 6.92 (d, J=8.4 Hz,
1H), 3.91 (s, 3H), 3.68 (s, 2H). .sup.13C NMR (100 MHz, CDCl.sub.3)
.delta. 154.8, 129.8, 127.3, 123.0, 122.7, 117.60, 112.4, 56.2,
22.4.
Example 15
2-(3-Fluoro-4-methoxyphenyl)acetonitrile
##STR00444##
[0291] To a suspension of t-BuOK (25.3 g, 0.207 mol) in THF (150
mL) was added a solution of TosMIC (20.3 g, 0.104 mol) in THF (50
mL) at -78.degree. C. The mixture was stirred for 15 minutes,
treated with a solution of 3-fluoro-4-methoxy-benzaldehyde (8.00 g,
51.9 mmol) in THF (50 mL) dropwise, and continued to stir for 1.5
hours at -78.degree. C. To the cooled reaction mixture was added
methanol (50 mL). The mixture was heated at reflux for 30 minutes.
Solvent of the reaction mixture was removed to give a crude
product, which was dissolved in water (200 mL). The aqueous phase
was extracted with EtOAc (100 mL.times.3). The combined organic
layers were dried and evaporated under reduced pressure to give
crude product, which was purified by column chromatography
(petroleum ether/ethyl acetate 10:1) to afford
2-(3-fluoro-4-methoxyphenyl)acetonitrile (5.0 g, 58%). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.02-7.05 (m, 2H), 6.94 (t, J=8.4 Hz,
1H), 3.88 (s, 3H), 3.67 (s, 2H). .sup.13C NMR (100 MHz, CDCl.sub.3)
.delta. 152.3, 147.5, 123.7, 122.5, 117.7, 115.8, 113.8, 56.3,
22.6.
Example 16
2-(4-Chloro-3-methoxyphenyl)acetonitrile
##STR00445##
[0292] Chloro-2-methoxy-4-methyl-benzene
[0293] To a solution of 2-chloro-5-methyl-phenol (93 g, 0.65 mol)
in CH.sub.3CN (700 mL) was added CH.sub.3I (110 g, 0.78 mol) and
K.sub.2CO.sub.3 (180 g, 1.3 mol). The mixture was stirred at
25.degree. C. overnight. The solid was filtered off and the
filtrate was evaporated under vacuum to give
1-chloro-2-methoxy-4-methyl-benzene (90 g, 89%). .sup.1H NMR (300
MHz, CDCl.sub.3) 87.22 (d, J=7.8 H, 1H), 6.74-6.69 (m 2H), 3.88 (s,
3H), 2.33 (s, 3H).
##STR00446##
4-Bromomethyl-1-chloro-2-methoxy-benzene
[0294] To a solution of 1-chloro-2-methoxy-4-methyl-benzene (50 g,
0.32 mol) in CCl.sub.4 (350 mL) was added NBS (57 g, 0.32 mol) and
AIBN (10 g, 60 mmol). The mixture was heated at reflux for 3 hours.
The solvent was evaporated under vacuum and the residue was
purified by column chromatography on silica gel (petroleum
ether/ethyl acetate a 20:1) to give
4-bromomethyl-1-chloro-2-methoxy-benzene (69 g, 92%). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.33-7.31 (m, 1H), 6.95-6.91 (m, 2H),
4.46 (s, 2H), 3.92 (s, 3H).
##STR00447##
2-(4-Chloro-3-methoxyphenyl)acetonitrile
[0295] To a solution of 4-bromomethyl-1-chloro-2-methoxy-benzene
(68.5 g, 0.290 mol) in C.sub.2HsOH (90%, 500 mL) was added NaCN
(28.5 g, 0.580 mol). The mixture was stirred at 60.degree. C.
overnight. Ethanol was evaporated and the residue was dissolved in
H.sub.2O. The mixture was extracted with ethyl acetate (300
mL.times.3). The combined organic layers were washed with brine,
dried over Na.sub.2SO.sub.4 and purified by column chromatography
on silica gel (petroleum ether/ethyl acetate 30:1) to give
2-(4-chloro-3-methoxyphenyl)acetonitrile (25 g, 48%). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.36 (d, J=8 Hz, 1H), 6.88-6.84 (m,
2H), 3.92 (s, 3H), 3.74 (s, 2H). .sup.13C NMR (100 MHz, CDCl.sub.3)
.delta. 155.4, 130.8, 129.7, 122.4, 120.7, 117.5, 111.5, 56.2,
23.5.
Example 17
1-(3-(Hydroxymethyl)-4-methoxyphenyl)cyclopropanecarboxylic
acid
##STR00448##
[0296] 1-(4-Methoxy-phenyl)-cyclopropanecarboxylic acid methyl
ester
[0297] To a solution of -(4-methoxy-phenyl)-cyclopropanecarboxylic
acid (50 g, 0.26 mol) in MeOH (500 mL) was added toluene-4-sulfonic
acid monohydrate (2.5 g, 13 mmol) at room temperature. The reaction
mixture was heated at reflux for 20 hours. MeOH was removed by
evaporation under vacuum and EtOAc (200 mL) was added. The organic
layer was washed with sat. aq. NaHCO.sub.3 (100 mL) and brine,
dried over anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum
to give 1-(4-methoxy-phenyl)-cyclopropanecarboxylic acid methyl
ester (53 g, 99%). .sup.1H NMR (CDC.sub.3, 400 MHz) .delta.
7.25-7.27 (m, 2H), 6.85 (d, J=8.8 Hz, 2H), 3.80 (s, 3H), 3.62 (s,
3H), 1.58 (m, 2H), 1.15 (m, 2H).
##STR00449##
1-(3-Chloromethyl-4-methoxy-phenyl)-cyclopropanecarboxylic acid
methyl ester
[0298] To a solution of 1-(4-methoxy-phenyl)-cyclopropanecarboxylic
acid methyl ester (30.0 g, 146 mmol) and MOMCl (29.1 g, 364 mmol)
in CS.sub.2 (300 mL) was added TiCl.sub.4 (8.30 g, 43.5 mmol) at
5.degree. C. The reaction mixture was heated at 30.degree. C. for 1
d and poured into ice-water. The mixture was extracted with
CH.sub.2Cl.sub.2 (150 mL.times.3). The combined organic extracts
were evaporated under vacuum to give
1-(3-chloromethyl-4-methoxy-phenyl)-cyclopropanecarboxylic acid
methyl ester (38.0 g), which was used in the next step without
further purification.
##STR00450##
1-(3-Hydroxymethyl-4-methoxy-phenyl)-cyclopropanecarboxylic acid
methyl ester
[0299] To a suspension of
1-(3-chloromethyl-4-methoxy-phenyl)-cyclopropanecarboxylic acid
methyl ester (20 g) in water (350 mL) was added Bu.sub.4NBr (4.0 g)
and Na.sub.2CO.sub.3 (90 g, 0.85 mol) at room temperature. The
reaction mixture was heated at 65.degree. C. overnight. The
resulting solution was acidified with aq. HCl (2 mol/L) and
extracted with EtOAc (200 mL.times.3). The organic layer was washed
with brine, dried over anhydrous Na.sub.2SO.sub.4 and evaporated
under vacuum to give crude product, which was purified by column
(petroleum ether/ethyl acetate 15:1) to give
1-(3-hydroxymethyl-4-methoxy-phenyl)-cyclopropanecarboxylic acid
methyl ester (8.0 g, 39%). .sup.1H NMR (CDC.sub.3, 400 MHz) .delta.
7.23-7.26 (m, 2H), 6.83 (d, J=8.0 Hz, 1H), 4.67 (s, 2H), 3.86 (s,
3H), 3.62 (s, 3H), 1.58 (q, J=3.6 Hz, 2H), 1.14-1.17 (m, 2H).
##STR00451##
1-[3-(tert-Butyl-dimethyl-silanyloxymethyl)-4-methoxy-phenyl]cyclopropane
carboxylic acid methyl ester
[0300] To a solution of
1-(3-hydroxymethyl-4-methoxy-phenyl)-cyclopropanecarboxylic acid
methyl ester (8.0 g, 34 mmol) in CH.sub.2Cl.sub.2 (100 mL) were
added imidazole (5.8 g, 85 mmol) and TBSCl (7.6 g, 51 mmol) at room
temperature. The mixture was stirred overnight at room temperature.
The mixture was washed with brine, dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under vacuum to give crude product,
which was purified by column (petroleum ether/ethyl acetate 30:1)
to give
1-[3-(tert-butyl-dimethyl-silanyloxymethyl)-4-methoxy-phenyl]-cyclopropan-
ecarboxylic acid methyl ester (6.7 g, 56%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.44-7.45 (m, 1H), 7.19 (dd, J=2.0,
8.4 Hz, 1H), 6.76 (d, J=8.4 Hz, 1H), 4.75 (s, 2H), 3.81 (s, 3H),
3.62 (s, 3H), 1.57-1.60 (m, 2H), 1.15-1.18 (m, 2H), 0.96 (s, 9H),
0.11 (s, 6H).
##STR00452##
1-(3-Hydroxymethyl-4-methoxy-phenyl)-cyclopropanecarboxylic
acid
[0301] To a solution of
1-[3-(tert-butyl-dimethyl-silanyloxymethyl)-4-methoxy-phenyl]-cyclopropan-
e carboxylic acid methyl ester (6.2 g, 18 mmol) in MeOH (75 mL) was
added a solution of LiOH.H.sub.2O (1.5 g, 36 mmol) in water (10 mL)
at 0.degree. C. The reaction mixture was stirred overnight at
40.degree. C. MeOH was removed by evaporation under vacuum. AcOH (1
mol/L, 40 mL) and EtOAc (200 mL) were added. The organic layer was
separated, washed with brine, dried over anhydrous Na.sub.2SO.sub.4
and evaporated under vacuum to provide
1-(3-hydroxymethyl-4-methoxy-phenyl)-cyclopropanecarboxylic acid
(5.3 g).
Example 18
2-(7-Chlorobenzo[d][1,3]dioxol-5-yl)acetonitrile
##STR00453##
[0302] 3-Chloro-4,5-dihydroxybenzaldehyde
[0303] To a suspension of 3-chloro-4-hydroxy-5-methoxy-benzaldehyde
(10 g, 54 mmol) in dichloromethane (300 mL) was added BBr.sub.3
(26.7 g, 107 mmol) dropwise at -40.degree. C. under N.sub.2. After
addition, the mixture was stirred at this temperature for 5 h and
then was poured into ice water. The precipitated solid was filtered
and washed with petroleum ether. The filtrate was evaporated under
reduced pressure to afford 3-chloro-4,5-dihydroxybenzaldehyde (9.8
g, 89%), which was directly used in the next step.
##STR00454##
7-Chlorobenzo[d][1,3]dioxole-S-carbaldehyde
[0304] To a solution of 3-chloro-4,5-dihydroxybenzaldehyde (8.0 g,
46 mmol) and BrClCH.sub.2 (23.9 g, 185 mmol) in dry DMF (100 mL)
was added Cs.sub.2CO.sub.3 (25 g, 190 mmol). The mixture was
stirred at 60.degree. C. overnight and was then poured into water.
The resulting mixture was extracted with EtOAc (50 mL.times.3). The
combined extracts were washed with brine (100 mL), dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to afford
7-chlorobenzo[d][1,3]dioxole-5-carbaldehyde (6.0 g, 70%). .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 9.74 (s, 1H), 7.42 (d, J=0.4 Hz,
1H), 7.26 (d, J=3.6 Hz, 1H), 6.15 (s, 2H).
##STR00455##
(7-Chlorobenzo[d][1,3]dioxol-5-yl)methanol
[0305] To a solution of 7-chlorobenzo[d][1,3]dioxole-5-carbaldehyde
(6.0 g, 33 mmol) in THF (50 mL) was added NaBH.sub.4 (2.5 g, 64
mmol)) in portions at 0.degree. C. The mixture was stirred at this
temperature for 30 min and then poured into aqueous NH.sub.4Cl
solution. The organic layer was separated, and the aqueous phase
was extracted with EtOAc (50 mL.times.3). The combined extracts
were dried over Na.sub.2SO.sub.4 and evaporated under reduced
pressure to afford (7-chlorobenzo[d][1,3]dioxol-5-yl)methanol,
which was directly used in the next step.
##STR00456##
4-Chloro-6-(chloromethyl)benzo[d][1,3]dioxole
[0306] A mixture of (7-chlorobenzo[d][1,3]-dioxol-5-yl)methanol
(5.5 g, 30 mmol) and SOCl.sub.2 (5.0 mL, 67 mmol) in
dichloromethane (20 mL) was stirred at room temperature for 1 h and
was then poured into ice water. The organic layer was separated and
the aqueous phase was extracted with dichloromethane (50
mL.times.3). The combined extracts were washed with water and
aqueous NaHCO.sub.3 solution, dried over Na.sub.2SO.sub.4 and
evaporated under reduced pressure to afford
4-chloro-6-(chloromethyl)benzo[d][1,3]dioxole, which was directly
used in the next step.
##STR00457##
2-(7-Chlorobenzo[d][1,3]dioxol-5-yl)acetonitrile
[0307] A mixture of 4-chloro-6-(chloromethyl)benzo[d][1,3]dioxole
(6.0 g, 29 mmol) and NaCN (1.6 g, 32 mmol) in DMSO (20 mL) was
stirred at 40.degree. C. for 1 h and was then poured into water.
The mixture was extracted with EtOAc (30 mL.times.3). The combined
organic layers were washed with water and brine, dried over
Na.sub.2SO.sub.4 and evaporated under reduced pressure to afford
2-(7-chlorobenzo[d][1,3]dioxol-5-yl)acetonitrile (3.4 g, 58%).
.sup.1H NMR .delta. 6.81 (s, 1H), 6.71 (s, 1H), 6.07 (s, 2H), 3.64
(s, 2H). .sup.13C-NMR 5149.2, 144.3, 124.4, 122.0, 117.4, 114.3,
107.0, 102.3, 23.1.
Example 19
1-(Benzo[d]oxazol-5-yl)cyclopropanecarboxylic acid
##STR00458##
[0308] 1-Benzooxazol-5-yl-cyclopropanecarboxylic acid methyl
ester
[0309] To a solution of
1-(3-amino-4-hydroxyphenyl)cyclopropanecarboxylic acid methyl ester
(3.00 g, 14.5 mmol) in DMF were added trimethyl orthoformate (5.30
g, 14.5 mmol) and a catalytic amount of p-toluenesulfonic acid
monohydrate (0.3 g) at room temperature. The mixture was stirred
for 3 hours at room temperature. The mixture was diluted with water
and extracted with EtOAc (100 mL.times.3). The combined organic
layers were dried over anhydrous Na.sub.2SO.sub.4 and evaporated
under vacuum to give 1-benzooxazol-5-yl-cyclopropanecarboxylic acid
methyl ester (3.1 g), which was directly used in the next step.
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.09 (s, 1), 7.75 (d,
J=1.2 Hz, 1H), 7.53-7.51 (m, 1H), 7.42-7.40 (m, 1H), 3.66 (s, 3H),
1.69-1.67 (m, 2H), 1.27-1.24 (m, 2H).
##STR00459##
1-(Benzo[d]oxazol-5-yl)cyclopropanecarboxylic acid
[0310] To a solution of 1-benzooxazol-5-yl-cyclopropanecarboxylic
acid methyl ester (2.9 g) in EtSH (30 mL) was added AlCl.sub.3 (5.3
g, 40 mmol) in portions at 0.degree. C. The reaction mixture was
stirred for 18 hours at room temperature. Water (20 mL) was added
dropwise at 0 DC. The resulting mixture was extracted with EtOAc
(100 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum to give the
crude product, which was purified by column chromatography on
silica gel (petroleum ether/ethyl acetate 1:2) to give
1-(benzo[d]oxazol-5-yl)cyclopropanecarboxylic acid (280 mg, 11%
over two steps). .sup.1H NMR (DMSO 400 MHz) .delta. 12.25 (brs,
1H), 8.71 (s, 1H), 7.70-7.64 (m, 2H), 7.40 (dd, J=1.6, 8.4 Hz, 1H),
1.49-1.46 (m, 2H), 1.21-1.18 (m, 2H). MS (ESI) m/e (M+H.sup.+)
204.4.
Example 20
2-(7-Fluorobenzo[d][1,3]dioxol-5-yl)acetonitrile
##STR00460##
[0311] 3-Fluoro-4,5-dihydroxy-benzaldehyde
[0312] To a suspension of 3-fluoro-4-hydroxy-5-methoxy-benzaldehyde
(1.35 g, 7.94 mmol) in dichloromethane (100 mL) was added BBr.sub.3
(1.5 mL, 16 mmol) dropwise at -78.degree. C. under N.sub.2. After
addition, the mixture was warmed to -30.degree. C. and it was
stirred at this temperature for 5 h. The reaction mixture was
poured into ice water. The precipitated solid was collected by
filtration and washed with dichloromethane to afford
3-fluoro-4,5-dihydroxy-benzaldehyde (1.1 g, 89%), which was
directly used in the next step.
##STR00461##
7-Fluoro-benzo[1,3]dioxole-5-carbaldehyde
[0313] To a solution of 3-fluoro-4,5-dihydroxy-benzaldehyde (1.5 g,
9.6 mmol) and BrClCH.sub.2 (4.9 g, 38.5 mmol) in dry DMF (50 mL)
was added Cs.sub.2CO.sub.3 (12.6 g, 39 mmol). The mixture was
stirred at 60.degree. C. overnight and was then poured into water.
The resulting mixture was extracted with EtOAc (50 mL.times.3). The
combined organic layers were washed with brine (100 mL), dried over
Na.sub.2SO.sub.4 and evaporated under reduced pressure to give the
crude product, which was purified by column chromatography on
silica gel (petroleum ether/ethyl acetate=10/1) to afford
7-fluoro-benzo[1,3]dioxole-5-carbaldehyde (0.80 g, 49%). .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 9.78 (d, J=0.9 Hz, 1H), 7.26 (dd,
J=1.5, 9.3 Hz, 1H), 7.19 (d, J=1.2 Hz, 1H), 6.16 (s, 2H).
##STR00462##
(7-Fluoro-benzo[1,3]dioxol-5-yl)-methanol
[0314] To a solution of 7-fluoro-benzo[1,3]dioxole-5-carbaldehyde
(0.80 g, 4.7 mmol) in MeOH (50 mL) was added NaBH.sub.4 (0.36 g,
9.4 mmol) in portions at 0.degree. C. The mixture was stirred at
this temperature for 30 min and was then concentrated to dryness.
The residue was dissolved in EtOAc. The EtOAc layer was washed with
water, dried over Na.sub.2SO.sub.4 and concentrated to dryness to
afford (7-fluoro-benzo[1,3]dioxol-5-yl)-methanol (0.80 g, 98%),
which was directly used in the next step.
##STR00463##
6-Chloromethyl-4-fluoro-benzo[1,3]dioxole
[0315] To SOCl.sub.2 (20 mL) was added
(7-fluoro-benzo[1,3]dioxol-5-yl)-methanol (0.80 g, 4.7 mmol) in
portions at 0.degree. C. The mixture was warmed to room temperature
over 1 h and then was heated at reflux for 1 h. The excess
SOCl.sub.2 was evaporated under reduced pressure to give the crude
product, which was basified with saturated aqueous NaHCO.sub.3 to
pH.about.7. The aqueous phase was extracted with EtOAc. (50
mL.times.3). The combined organic layers were dried over
Na.sub.2SO.sub.4 and evaporated under reduced pressure to give
6-chloromethyl-4-fluoro-benzo[1,3]dioxole (0.80 g, 92%), which was
directly used in the next step.
##STR00464##
2-(7-Fluorobenzo[d[(1,3]dioxol-5-yl)acetonitrile
[0316] A mixture of 6-chloromethyl-4-fluoro-benzo[1,3]dioxole (0.80
g, 4.3 mmol) and NaCN (417 mg, 8.51 mmol) in DMSO (20 mL) was
stirred at 30.degree. C. for 1 h and was then poured into water.
The mixture was extracted with EtOAc (50 mL.times.3). The combined
organic layers were washed with water (50 mL) and brine (50 mL),
dried over Na.sub.2SO.sub.4 and evaporated under reduced pressure
to give the crude product, which was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate=10/1)
to afford 2-(7-fluorobenzo[d][1,3]dioxol-5-yl)acetonitrile (530 mg,
70%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 6.68-6.64 (m, 2H),
6.05 (s, 2H), 3.65 (s, 2H). .sup.13C-NMR 8151.1, 146.2, 134.1,
124.2, 117.5, 110.4, 104.8, 102.8, 23.3.
Example 21
1-(1H-Indol-5-yl)cyclopropanecarboxylic acid
##STR00465##
[0317] Methyl 1-phenylcyclopropanecarboxylate
[0318] To a solution of 1-phenylcyclopropanecarboxylic acid (25 g,
0.15 mol) in CH.sub.3OH (200 mL) was added TsOH (3 g, 0.1 mol) at
room temperature. The mixture was refluxed overnight. The solvent
was evaporated under reduced pressure to give crude product, which
was dissolved into EtOAc. The EtOAc layer was washed with aq. sat.
NaHCO.sub.3. The organic layer was dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under reduced pressure to give
methyl 1-phenylcyclopropanecarboxylate (26 g, 96%), which was used
directly in the next step. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.37-7.26 (m, 5H), 3.63 (s, 3H), 1.63-1.60 (m, 2H),
1.22-1.19 (m, 2H).
##STR00466##
Methyl 1-(4-nitrophenyl)cyclopropanecarboxylate
[0319] To a solution of 1-phenylcyclopropanecarboxylate (20.62 g,
0.14 mol) in H.sub.2SO.sub.4/CH.sub.2Cl.sub.2 (40 mL/40 mL) was
added KNO.sub.3 (12.8 g, 0.13 mol) in portion at 0.degree. C. The
mixture was stirred for 0.5 hr at 0.degree. C. Ice water was added
and the mixture was extracted with EtOAc (100 mL.times.3). The
organic layers were dried with anhydrous Na.sub.2SO.sub.4 and
evaporated to give methyl 1-(4-nitrophenyl)cyclopropanecarboxylate
(21 g, 68%), which was used directly in the next step. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 8.18 (dd, J=2.1, 6.9 Hz, 2H), 7.51
(dd. J=2.1, 6.9 Hz, 2H), 3.64 (s, 3H), 1.72-1.69 (m, 2H), 1.25-1.22
(m, 2H).
##STR00467##
Methyl 1-(4-aminophenyl)cyclopropanecarboxylate
[0320] To a solution of methyl
1-(4-nitrophenyl)cyclopropanecarboxylate (20 g, 0.09 mol) in MeOH
(400 mL) was added Ni (2 g) under nitrogen atmosphere. The mixture
was stirred under hydrogen atmosphere (1 atm) at room temperature
overnight. The catalyst was filtered off through a pad of Celite
and the filtrate was evaporated under vacuum to give crude product,
which was purified by chromatography column on silica gel
(petroleum ether/ethyl acetate=10:1) to give methyl
1-(4-aminophenyl)cyclopropanecarboxylate (11.38 g, 66%). .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 7.16 (d, J=8.1 Hz, 2H), 6.86 (d,
J=7.8 Hz, 2H), 4.31 (br, 2H), 3.61 (s, 3H), 1.55-1.50 (m, 2H),
1.30-1.12 (m, 2H).
##STR00468##
Methyl 1-(4-amino-3-bromophenyl)cyclopropanecarboxylate
[0321] To a solution of methyl
1-(4-aminophenyl)cyclopropanecarboxylate (10.38 g, 0.05 mol) in
acetonitrile (200 mL) was added NBS (9.3 g, 0.05 mol) at room
temperature. The mixture was stirred overnight. Water (200 mL) was
added. The organic layer was separated and the aqueous phase was
extracted with EtOAc (80 mL.times.3). The organic layers were dried
with anhydrous Na.sub.2SO.sub.4 and evaporated to give methyl
1-(4-amino-3-bromophenyl)cyclopropanecarboxylate (10.6 g, 78%),
which was used directly in the next step. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.38 (d, J=2.0 Hz, 1H), 7.08 (dd, J=1.6, 8.4
Hz, 1H), 6.70 (d, J=8.4 Hz, 1H), 3.62 (s, 3H), 1.56-1.54 (m, 2H),
1.14-1.11 (m, 2H).
##STR00469##
Methyl 1-(4-amino-3-((trimethylsilyl)ethynyl)phenyl)cyclopropane
carboxylate
[0322] To a degassed solution of methyl
1-(4-amino-3-bromophenyl)cyclopropane carboxylate (8 g, 0.03 mol)
in Et.sub.3N (100 mL) was added ethynyl-trimethyl-silane (30 g, 0.3
mol), DMAP (5% mol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (5% mol) under
N.sub.2. The mixture was refluxed at 70.degree. C. overnight. The
insoluble solid was filtered off and washed with EtOAc (100
mL.times.3). The filtrate was evaporated under reduced pressure to
give a residue, which was purified by chromatography column on
silica gel (petroleum ether/ethyl acetate=20:1) to give methyl
1-(4-amino-3-((trimethylsilyl)ethynyl)phenyl)cyclopropanecarboxylate
(4.8 g, 56%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.7.27 s, 1H),
7.10 (dd, J=2.1, 8.4 Hz, 1H), 6.64 (d, J=8.4 Hz, 1H), 3.60 (s, 3H),
1.55-1.51 (m, 2H), 1.12-1.09 (m, 2H), 0.24 (s, 9H).
##STR00470##
Methyl 1-(1H-indol-5-yl)cyclopropanecarboxylate
[0323] To a degassed solution of methyl
1-(4-amino-3-((trimethylsilyl)ethynyl)phenyl)
cyclopropanecarboxylate (4.69 g, 0.02 mol) in DMF (20 mL) was added
CuI (1.5 g, 0.008 mol) under N.sub.2 at room temperature. The
mixture was stirred for 3 hr at room temperature. The insoluble
solid was filtered off and washed with EtOAc (50 mL.times.3). The
filtrate was evaporated under reduced pressure to give a residue,
which was purified by chromatography column on silica gel
(petroleum ether/ethyl acetate=20:1) to give methyl
1-(1H-indol-5-yl)cyclopropanecarboxylate (2.2 g, 51%). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.61 (s, 1H), 7.33 (d, J=8.4 Hz, 1H),
7.23-7.18 (m, 2H), 6.52-6.51 (m, 1H) 3.62 (s, 3H), 1.65-1.62 (m,
2H), 1.29-1.23 (m, 2H).
##STR00471##
1-(1H-Indol-5-yl)cyclopropanecarboxylic acid
[0324] To a solution of methyl
1-(1H-indol-5-yl)cyclopropanecarboxylate (1.74 g, 8 mmol) in
CH.sub.3OH (50 m L) and water (20 mL) was added LiOH (1.7 g, 0.04
mol). The mixture was heated at 45.degree. C. for 3 hr. Water was
added and the mixture was acidified with concentrated HCl to
pH.about.3 before being extracted with EtOAc (20 mL.times.3). The
organic layers were dried over anhydrous Na.sub.2SO.sub.4 and
evaporated to give 1-(1H-indol-5-yl)cyclopropanecarboxylic acid
(1.4 g, 87%). .sup.1H NMR (300 MHz, DMSO-d.sub.6) 7.43 (s, 1H),
7.30-7.26 (m, 2H), 7.04 (dd, J=1.5, 8.4 Hz, 1H), 6.35 (s, 1H),
1.45-1.41 (m, 2H), 1.14-1.10 (m, 2H).
Example 22
1-(4-Oxochroman-6-yl)cyclopropanecarboxylic acid
##STR00472##
[0325]
1-[4-(2-tert-Butoxycarbonyl-ethoxy)-phenyl]-cyclopropanecarboxylic
methyl ester
[0326] To a solution of 1-(4-hydroxy-phenyl)-cyclopropanecarboxylic
methyl ester (7.0 g, 3.6 mmol) in acrylic tert-butyl ester (50 mL)
was added Na (42 mg, 1.8 mmol) at room temperature. The mixture was
heated at 110.degree. C. for 1 h. After cooling to room
temperature, the resulting mixture was quenched with water and
extracted with EtOAc (100 mL.times.3). The combined organic
extracts were dried over anhydrous Na.sub.2SO.sub.4 and evaporated
under vacuum to give the crude product, which was purified by
column chromatography on silica gel (petroleum ether/ethyl acetate
20:1) to give
1-[4-(2-tert-butoxycarbonyl-ethoxy)-phenyl]-cyclopropanecarboxylic
methyl ester (6.3 g, 54%) and unreacted start material (3.0 g).
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.24 (d, J=8.7 Hz, 2H),
6.84 (d, J=8.7 Hz, 2H), 4.20 (t, J=6.6 Hz, 2H), 3.62 (s, 3H), 2.69
(t, J=6.6 Hz, 2H), 1.59-1.56 (m, 2H), 1.47 (s, 9H), 1.17-1.42 (m,
2H).
##STR00473##
1-[4-(2-Carboxy-ethoxy)-phenyl-cyclopropanecarboxylic methyl
ester
[0327] A solution of
1-[4-(2-tert-butoxycarbonyl-ethoxy)-phenyl]-cyclopropanecarboxylic
methyl ester (6.3 g, 20 mmol) in HCl (20%, 200 mL) was heated at
110.degree. C. for 1 h. After cooling to room temperature, the
resulting mixture was filtered. The solid was washed with water and
dried under vacuum to give
1-[4-(2-carboxy-ethoxy)-phenyl]-cyclopropanecarboxylic methyl ester
(5.0 g, 96%). .sup.1H NMR (300 MHz, DMSO) .delta. 7.23-7.19 (m,
2H), 6.85-6.81 (m, 2H), 4.13 (t, J=6.0 Hz, 2H), 3.51 (s, 3H), 2.66
(t, J=6.0 Hz, 2H), 1.43-1.39 (m, 2H), 1.14-1.10 (m, 2H).
##STR00474##
1-(4-Oxochroman-6-yl)cyclopropanecarboxylic acid
[0328] To a solution of
1-[4-(2-carboxy-ethoxy)-phenyl]-cyclopropanecarboxylic methyl ester
(5.0 g, 20 mmol) in CH.sub.2Cl.sub.2 (50 mL) were added oxalyl
chloride (4.8 g, 38 mmol) and two drops of DMF at 0.degree. C. The
mixture was stirred at 0-5.degree. C. for 1 h and then evaporated
under vacuum. To the resulting mixture was added CH.sub.2Cl.sub.2
(50 mL) at 0.degree. C. and stirring was continued at 0-5.degree.
C. for 1 h. The reaction was slowly quenched with water and was
extracted with EtOAc (50 mL.times.3). The combined organic extracts
were dried over anhydrous Na.sub.2SO.sub.4 and evaporated under
vacuum to give the crude product, which was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate
20:1-2:1) to give 1-(4-oxochroman-6-yl)cyclopropanecarboxylic acid
(830 mg, 19%) and methyl
1-(4-oxochroman-6-yl)cyclopropanecarboxylate (1.8 g, 38%).
1-(4-Oxochroman-6-yl)cyclopropane-carboxylic acid: .sup.1H NMR (400
MHz, DMSO) .delta. 12.33 (br s, 1H), 7.62 (d, J=2.0 Hz, 1H), 7.50
(dd, J=2.4, 8.4 Hz, 1H), 6.95 (d, J=8.4 Hz, 1H), 4.50 (t, J=6.4 Hz,
2H), 2.75 (t, J=6.4 Hz, 2H), 1.44-1.38 (m, 2H), 1.10-1.07 (m, 2H).
MS (ESI) m/z (M+H.sup.+) 231.4.
1-(4-Oxochroman-6-yl)cyclopropanecarboxylate: .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.83 (d, J=2.4 Hz, 1H), 7.48 (dd, J=2.4, 8.4
Hz, 1H), 6.93 (d, J=8.4 Hz, 1H), 4.55-4.52 (m, 2H), 3.62 (s, 3H),
2.80 (t, J=6.4 Hz, 2H), 1.62-1.56 (m, 2H), 1.18-1.15 (m, 2H).
Example 23
1-(4-Hydroxy-4-methoxychroman-6-yl)cyclopropanecarboxylic acid
##STR00475##
[0329] 1-(4-Hydroxy-4-methoxycroman-6-yl)cyclopropanecarboxylic
acid
[0330] To a solution of methyl
1-(4-oxochroman-6-yl)cyclopropanecarboxylate (1.0 g, 4.1 mmol) in
MeOH (20 mL) and water (20 mL) was added LiOH.H.sub.2O (0.70 g, 16
mmol) in portions at room temperature. The mixture was stirred
overnight at room temperature before the MeOH was removed by
evaporation under vacuum. Water and Et.sub.2O were added to the
residue and the aqueous layer was separated, acidified with HCl and
extracted with EtOAc (50 mL.times.3). The combined organic extracts
dried over anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum
to give 1-(4-hydroxy-4-methoxychroman-6-yl)cyclopropanecarboxylic
acid (480 mg, 44%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 12.16
(s, 1H), 7.73 (d, J=2.0 Hz, 1H), 7.47 (dd, J=2.0, 8.4 Hz, 1H), 6.93
(d, J=8.8 Hz, 1H), 3.83-3.80 (m, 2H), 3.39 (s, 3H), 3.28-3.25 (m,
2H), 1.71-1.68 (m, 2H), 1.25-1.22 (m, 2H). MS (ESI) m/z (M+H.sup.+)
263.1.
Example 24
1-(4-Hydroxy-4-methoxychroman-6-yl)cyclopropanecarboxylic acid
##STR00476##
[0331] 1-Chroman-6-yl-cyclopropanecarboxylic methyl ester
[0332] To trifluoroacetic acid (20 mL) was added NaBH.sub.4 (0.70
g, 130 mmol) in portions at 0.degree. C. under N.sub.2 atmosphere.
After stirring for 5 min, a solution of
1-(4-oxo-chroman-6-yl)-cyclopropanecarboxylic methyl ester (1.6 g,
6.5 mmol) was added at 15.degree. C. The reaction mixture was
stirred for 1 h at room temperature before being slowly quenched
with water. The resulting mixture was extracted with EtOAc (50
mL.times.3). The combined organic extracts dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under vacuum to give
1-chroman-6-yl-cyclopropanecarboxylic methyl ester (1.4 g, 92%),
which was used directly in the next step. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.07-7.00 (m, 2H), 6.73 (d, J=8.4 Hz, 1H), 4.17
(t, J=5.1 Hz, 2H), 3.62 (s, 3H), 2.79-2.75 (nm, 2H), 2.05-1.96 (m,
2H), 1.57-1.54 (m, 2H), 1.16-1.13 (m, 2H).
##STR00477##
1-(4-Hydroxy-4methoxychroman-6-yl)cyclopropanecarboxylic acid
[0333] To a solution of 1-chroman-6-yl-cyclopropanecarboxylic
methyl ester (1.4 g, 60 mmol) in MeOH (20 mL) and water (20 mL) was
added LiOH.H.sub.2O (1.0 g, 240 mmol) in portions at room
temperature. The mixture was stirred overnight at room temperature
before the MeOH was removed by evaporation under vacuum. Water and
Et.sub.2O were added and the aqueous layer was separated, acidified
with HCl and extracted with EtOAc (50 ml.times.3). The combined
organic extracts dried over anhydrous Na.sub.2SO.sub.4 and
evaporated under vacuum to give
1-(4-Hydroxy-4-methoxychroman-6-yl)cyclopropanecarboxylic acid (1.0
g, 76%). .sup.1H NMR (400 MHz, DMSO) .delta. 12.10 (br s, 1H), 6.95
(d, J=2.4 Hz, 2H), 6.61-6.59 (m, 1H), 4.09-4.06 (m, 2H), 2.70-2.67
(m, 2H), 1.88-1.86 (m, 2H), 1.37-1.35 (m, 2H), 1.04-1.01 (m, 2H).
MS (ESI) m/z (M+H.sup.+) 217.4.
Example 25
1-(3-Methylbenzo[d]isoxazol-5-yl)cyclopropanecarboxylic acid
##STR00478##
[0334] 1-(3-Acetyl-4-hydroxy-phenyl)-cyclopropanecarboxylic methyl
ester
[0335] To a stirred suspension of AlCl.sub.3 (58 g, 440 mmol) in
CS.sub.2 (500 mL) was added acetyl chloride (7.4 g, 95 mmol) at
room temperature. After stirring for 5 min, methyl
1-(4-methoxyphenyl)cyclopropanecarboxylate (15 g, 73 mmol) was
added. The reaction mixture was heated at reflux for 2 h before ice
water was added carefully to the mixture at room temperature. The
resulting mixture was extracted with EtOAc (150 mL.times.3). The
combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under reduced pressure to give
1-(3-acetyl-4-hydroxy-phenyl)-cyclopropanecarboxylic methyl ester
(15 g, 81%), which was used in the next step without further
purification. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 12.28 (s,
1H), 7.67 (d, J=2.0 Hz, 1H), 7.47 (dd, J=2.0, 8.4 Hz, 1H), 6.94 (d,
J=8.4 Hz, 1H), 3.64 (s, 3H), 2.64 (s, 3H), 1.65-1.62 (m, 2H),
1.18-1.16 (m, 2H).
##STR00479##
1-[4-Hydroxy-3-(1-hydroxyimino-ethyl)-phenyl]-cyclopropanecarboxylic
methyl ester
[0336] To a stirred solution of
1-(3-acetyl-4-hydroxy-phenyl)-cyclopropanecarboxylic methyl ester
(14.6 g, 58.8 mmol) in EtOH (500 mL) were added hydroxylamine
hydrochloride (9.00 g, 129 mmol) and sodium acetate (11.6 g, 141
mmol) at room temperature. The resulting mixture was heated at
reflux overnight. After removal of EtOH under vacuum, water (200
mL) and EtOAc (200 mL) were added. The organic layer was separated
and the aqueous layer was extracted with EtOAc (100 mL.times.3).
The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under vacuum to give
1-[4-hydroxy-3-(1-hydroxyimino-ethyl)-phenyl]-cyclopropanecarboxylic
methyl ester (14.5 g, 98%), which was used in the next step without
further purification. .sup.1H NMR (CDC.sub.3, 400 MHz) .delta.
11.09 (s, 1H), 7.39 (d, J=2.0 Hz, 1H), 7.23 (d, J=2.0 Hz, 1H), 7.14
(s, 1H), 6.91 (d, J=8.4 Hz, 1H), 3.63 (s, 3H), 2.36 (s, 3H),
1.62-1.59 (m, 2H), 1.18-1.15 (m, 2H).
##STR00480##
(E)-Methyl
1-(3-(1-(acetoxyimino)ethyl)-4-hydroxyphenyl)cyclopropane
carboxylate
[0337] The solution of
1-[4-hydroxy-3-(1-hydroxyimino-ethyl)-phenyl]-cyclopropanecarboxylic
methyl ester (10.0 g, 40.1 mmol) in Ac.sub.2O (250 mL) was heated
at 45.degree. C. for 4 h. The Ac.sub.2O was removed by evaporation
under vacuum before water (100 mL) and EtOAc (100 mL) were added.
The organic layer was separated and the aqueous layer was extracted
with EtOAc (100 mL.times.2). The combined organic layers were dried
over anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum to give
(E)-methyl
1-(3-(1-(acetoxyimino)ethyl)-4-hydroxyphenyl)cyclopropanecarboxylate
(10.5 g, 99%), which was used in the next step without further
purification.
##STR00481##
Methyl 1-(3-methylbenzo[d]isoxazol-5-yl)cyclopropanecarboxylate
[0338] A solution of (E)-methyl
1-(3-(1-(acetoxyimino)ethyl)-4-hydroxyphenyl)cyclopropane
carboxylate (10.5 g, 39.6 mmol) and pyridine (31.3 g, 396 mmol) in
DMF (150 mL) was heated at 125.degree. C. for 10 h. The cooled
reaction mixture was poured into water (250 mL) and was extracted
with EtOAc (100 mL.times.3). The combined organic layers were dried
over anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum to give
the crude product, which was purified by column chromatography on
silica gel (petroleum ether/ethyl acetate 50:1) to give methyl
1-(3-methylbenzo[d]isoxazol-5-yl)cyclopropanecarboxylate (7.5 g,
82%). .sup.1H NMR (CDCl.sub.3 300 MHz) .delta. 7.58-7.54 (m, 2H),
7.48 (dd, J=1.5, 8.1 Hz, 1H), 3.63 (s, 3H), 2.58 (s, 3H), 1.71-1.68
(m, 2H), 1.27-1.23 (m, 2H).
##STR00482##
1-(3-Methylbenzo[d]isoxazol-5-yl)cyclopropanecarboxylic acid
[0339] To a solution of methyl
1-(3-methylbenzo[d]isoxazol-5-yl)cyclopropanecarboxylate (1.5 g,
6.5 mmol) in MeOH (20 mL) and water (2 mL) was added LiOH.H.sub.2O
(0.80 g, 19 mmol) in portions at room temperature. The reaction
mixture was stirred at room temperature overnight before the MeOH
was removed by evaporation under vacuum. Water and Et.sub.2O were
added and the aqueous layer was separated, acidified with HCl and
extracted with EtOAc (50 mL.times.3). The combined organic extracts
were dried over anhydrous Na.sub.2SO.sub.4 and evaporated under
vacuum to give
1-(3-methylbenzo[d]isoxazol-5-yl)cyclopropanecarboxylic acid (455
mg, 32%). .sup.1H NMR (400 MHz, DMSO) .delta. 12.40 (br s, 1H),
7.76 (s, 1H), 7.60-7.57 (m, 2H), 2.63 (s, 3H), 1.52-1.48 (m, 2H),
1.23-1.19 (m, 2H). MS (ESI) m/z (M+H.sup.+) 218.1.
Example 26
1-(Spiro[benzo[d][1,3]dioxole-2,1'-cyclobutane]-5-yl)cyclopropane
carboxylic acid
##STR00483##
[0340] 1-(3,4-Dihydroxy-phenyl)-cyclopropanecarboxylic methyl
ester
[0341] To a solution of
1-(3,4-dihydroxyphenyl)cyclopropanecarboxylic acid (4.5 g) in MeOH
(30 mL) was added TsOH (0.25 g, 1.3 mmol). The stirring was
continued at 50.degree. C. overnight before the mixture was cooled
to room temperature. The mixture was concentrated under vacuum and
the residue was purified by column chromatography on silica gel
(petroleum ether/ethyl acetate 3:1) to give
1-(3,4-dihydroxy-phenyl)-cyclopropanecarboxylic methyl ester (2.1
g). .sup.1H NMR (DMSO 300 MHz) .delta. 8.81 (brs, 2H), 6.66 (d,
J=2.1 Hz, 1H), 6.61 (d, J=8.1 Hz, 1H), 6.53 (dd, J=2.1, 8.1 Hz,
1H), 3.51 (s, 3H), 1.38-1.35 (m, 2H), 1.07-1.03 (m, 2H).
##STR00484##
Methyl
1-(spiro[benzo[d][1,3]dioxole-2,1'-cyclobutane]-5-yl)cyclopropane
carboxylate
[0342] To a solution of
1-(3,4-dihydroxy-phenyl)-cyclopropanecarboxylic methyl ester (1.0
g, 4.8 mmol) in toluene (30 mL) was added TsOH (0.10 g, 0.50 mmol)
and cyclobutanone (0.70 g, 10 mmol). The reaction mixture was
heated at reflux for 2 h before being concentrated under vacuum.
The residue was purified by chromatography on silica gel (petroleum
ether/ethyl acetate 15:1) to give methyl
1-(spiro[benzo[d][1,3]dioxole-2,1'-cyclobutane]-5-yl)cyclopropanecarboxyl-
ate (0.6 g, 50%). .sup.1H NMR (CDCl.sub.3 300 MHz) .delta.
6.78-6.65 (m, 3H), 3.62 (s, 3H), 2.64-2.58 (m, 4H), 1.89-1.78 (m,
2H), 1.56-1.54 (m, 2H), 1.53-1.12 (m, 2H).
##STR00485##
1-(Spiro[benzo[d][1,3]dioxole-2,1'-cyclobutane]-5-yl)cyclopropane
carboxylic acid
[0343] To a mixture of methyl
1-(spiro[benzo[d][1,3]dioxole-2,1'-cyclobutane]-5-yl)cyclopropanecarboxyl-
ate (0.60 g, 2.3 mmol) in THF/H.sub.2O (4:1, 10 mL) was added LiOH
(0.30 g, 6.9 mmol). The mixture was stirred at 60.degree. C. for 24
h. HCl (0.5 N) was added slowly to the mixture at 0.degree. C.
until pH 2-3. The mixture was extracted with EtOAc (10 mL.times.3).
The combined organic phases were washed with brine, dried over
anhydrous MgSO.sub.4, and washed with petroleum ether to give
1-(spiro[benzo[d][1,3]-dioxole-2,1'-cyclobutane]-5-yl)cyclopropane
carboxylic acid (330 mg, 59%). .sup.1HNMR (400 MHz, CDCl.sub.3)
.delta. 6.78-6.65 (m, 3H), 2.65-2.58 (m, 4H), 1.86-1.78 (m, 2H),
1.63-1.60 (m, 2H), 1.26-1.19 (m, 2H);
Example 27
2-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)acetonitrile
##STR00486##
[0344] 2,3-Dihydro-benzo[1,4]dioxine-6-carboxylic acid ethyl
ester
[0345] To a suspension of Cs.sub.2CO.sub.3 (270 g, 1.49 mol) in DMF
(1000 mL) were added 3,4-dihydroxybenzoic acid ethyl ester (54.6 g,
0.3 mol) and 1,2-dibromoethane (54.3 g, 0.29 mol) at room
temperature. The resulting mixture was stirred at 80.degree. C.
overnight and then poured into ice-water. The mixture was extracted
with EtOAc (200 mL.times.3). The combined organic layers were
washed with water (200 mL.times.3) and brine (100 mL), dried over
Na.sub.2SO.sub.4 and concentrated to dryness. The residue was
purified by column (petroleum ether/ethyl acetate 50:1) on silica
gel to obtain 2,3-dihydro-benzo[1,4]dioxine-6-carboxylic acid ethyl
ester (18 g, 29%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.53
(dd, J=1.8, 7.2 Hz, 2H), 6.84-6.87 (m, 1H), 4.22-4.34 (m, 6H), 1.35
(t, J=7.2 Hz, 3H).
##STR00487##
(2,3-Dihydro-benzo[1,4]dioxin-6-yl)-methanol
[0346] To a suspension of LiAlH.sub.4 (2.8 g, 74 mmol) in THF (20
mL) was added dropwise a solution of
2,3-dihydro-benzo[1,4]dioxine-6-carboxylic acid ethyl ester (15 g,
72 mmol) in THF (10 mL) at 0.degree. C. under N.sub.2. The mixture
was stirred at room temperature for 1 h and then quenched carefully
with addition of water (2.8 mL) and NaOH (10%, 28 mL) with cooling.
The precipitated solid was filtered off and the filtrate was
evaporated to dryness to obtain
(2,3-dihydro-benzo[1,4]dioxin-6-yl)-methanol (10.6 g). .sup.1H NMR
(300 MHz, DMSO-d.sub.6) .delta. 6.73-6.78 (m, 3H), 5.02 (t, J=5.7
Hz, 1H), 4.34 (d, J=6.0 Hz, 2H), 4.17-4.20 (m, 4H).
##STR00488##
6-Chloromethyl-2,3-dihydro-benzo[1,4]dioxine
[0347] A mixture of (2,3-dihydro-benzo[1,4]dioxin-6-yl)methanol
(10.6 g) in SOCl.sub.2 (10 mL) was stirred at room temperature for
10 min and then poured into ice-water. The organic layer was
separated and the aqueous phase was extracted with dichloromethane
(50 mL.times.3). The combined organic layers were washed with
NaHCO.sub.3 (sat solution), water and brine, dried over
Na.sub.2SO.sub.4 and concentrated to dryness to obtain
6-chloromethyl-2,3-dihydro-benzo[1,4]dioxine (12 g, 88% over two
steps), which was used directly in next step.
##STR00489##
2-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)acetonitrile
[0348] A mixture of 6-chloromethyl-2,3-dihydro-benzo[1,4]dioxine
(12.5 g, 67.7 mmol) and NaCN (4.30 g, 87.8 mmol) in DMSO (50 mL)
was stirred at rt for 1 h. The mixture was poured into water (150
mL) and then extracted with dichloromethane (50 mL.times.4). The
combined organic layers were washed with water (50 mL.times.2) and
brine (50 mL), dried over Na.sub.2SO.sub.4 and concentrated to
dryness. The residue was purified by column (petroleum ether/ethyl
acetate 50:1) on silica gel to obtain
2-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)acetonitrile as a yellow oil
(10.2 g, 86%). .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta. 6.78-6.86
(m, 3H), 4.25 (s, 4H), 3.63 (s, 2H).
[0349] The following Table 2 contains a list of carboxylic acid
building blocks that were commercially available, or prepared by
one of the three methods described above:
TABLE-US-00002 TABLE 2 Carboxylic acid building blocks. Name
Structure 1-benzo[1,3]dioxol-5-ylcyclopropane-1- carboxylic acid
##STR00490## 1-(2,2-difluorobenzo[1,3]dioxol-5-
yl)cyclopropane-1-carboxylic acid ##STR00491## 1-(3,4-
dihydroxyphenyl)cyclopropanecarboxylic acid ##STR00492##
1-(3-methoxyphenyl)cyclopropane-1- carboxylic acid ##STR00493##
1-(2-methoxyphenyl)cyclopropane-1- carboxylic acid ##STR00494##
1-[4- (trifluoromethoxy)phenyl]cyclopropane- 1-carboxylic acid
##STR00495## 1-(2,2-dimethylbenzo[d][1,3]dioxol-5-
yl)cyclopropanecarboxylic acid ##STR00496##
tetrahydro-4-(4-methoxyphenyl)-2H- pyran-4-carboxylic acid
##STR00497## 1-phenylcyclopropane-1-carboxylic acid ##STR00498##
1-(4-methoxyphenyl)cyclopropane-1- carboxylic acid ##STR00499##
1-(4-chlorophenyl)cyclopropane-1- carboxylic acid ##STR00500##
1-(3- hydroxyphenyl)cyclopropanecarboxylic acid ##STR00501##
1-phenylcyclopentanecarboxylic acid ##STR00502##
1-(2-oxo-2,3-dihydrobenzo[d]oxazol-5- yl)cyclopropanecarboxylic
acid ##STR00503## 1-(benzofuran-5- yl)cyclopropanecarboxylic acid
##STR00504## 1-(4- methoxyphenyl)cyclohexanecarboxylic acid
##STR00505## 1-(4-chlorophenyl)cyclohexanecarboxylic acid
##STR00506## 1-(2,3-dihydrobenzofuran-5- yl)cyclopropanecarboxylic
acid ##STR00507## 1-(3,3-dimethyl-2,3-dihydrobenzofuran-
5-yl)cyclopropanecarboxylic acid ##STR00508##
1-(7-methoxybenzo[d][1,3]dioxol-5- yl)cyclopropanecarboxylic acid
##STR00509## 1-(3-hydroxy-4- methoxyphenyl)cyclopropanecarboxylic
acid ##STR00510## 1-(4-chloro-3-
hydroxyphenyl)cyclopropanecarboxylic acid ##STR00511##
1-(3-(benzyloxy)-4- chlorophenyl)cyclopropanecarboxylic acid
##STR00512## 1-(4- chlorophenyl)cyclopentanecarboxylic acid
##STR00513## 1-(3-(benzyloxy)-4-
methoxyphenyl)cyclopropanecarboxylic acid ##STR00514##
1-(3-chloro-4- methoxyphenyl)cyclopropanecarboxylic acid
##STR00515## 1-(3-fluoro-4- methoxyphenyl)cyclopropanecarboxylic
acid ##STR00516## 1-(4-methoxy-3-
methylphenyl)cyclopropanecarboxylic acid ##STR00517##
1-(4-(benzyloxy)-3- methoxyphenyl)cyclopropanecarboxylic acid
##STR00518## 1-(4-chloro-3- methoxyphenyl)cyclopropanecarboxylic
acid ##STR00519## 1-(3-chloro-4-
hydroxypheny)cyclopropanecarboxylic acid ##STR00520##
1-(3-(hydroxymethyl)-4- methoxyphenyl)cyclopropanecarboxylic acid
##STR00521## 1-(4- methoxyphenyl)cyclopentanecarboxylic acid
##STR00522## 1-phenylcyclobexanecarboxylic acid ##STR00523##
1-(3,4- dimethoxyphenyl)cyclopropanecarboxylic acid ##STR00524##
1-(7-chlorobenzo[d][1,3]dioxol-5- yl)cyclopropanecarboxylic acid
##STR00525## 1-(benzo[d]oxazol-5- yl)cyclopropanecarboxylic acid
##STR00526## 1-(7-fluorobenzo[d][1,3]dioxol-5-
yl)cyclopropanecarboxylic acid ##STR00527## 1-(3,4-
difluorophenyl)cyclopropanecarboxylic acid ##STR00528##
1-(1H-indol-5-yl)cyclopropanecarboxylic acid ##STR00529##
1-(1H-benzo[d]imidazol-5- yl)cyclopropanecarboxylic acid
##STR00530## 1-(2-methyl-1H-benzo[d]imidazol-5-
yl)cyclopropanecarboxylic acid ##STR00531##
1-(1-methyl-1H-benzo[d]imidazol-5- yl)cyclopropanecarboxylic acid
##STR00532## 1-(3-methylbenzo[d]isoxazol-5-
yl)cyclopropanecarboxylic acid ##STR00533##
1-(spiro[benzo[d][1,3]dioxole-2,1'- cyclobutane]-5-
yl)cyclopropanecarboxylic acid ##STR00534##
1-(1H-benzo[d][1,2,3]triazol-5- yl)cyclopropanecarboxylic acid
##STR00535## 1-(1-methyl-1H-benzo[d][1,2,3]triazol-5-
yl)cyclopropanecarboxylic acid ##STR00536##
1-(1,3-dihydroisobenzofuran-5- yl)cyclopropanecarboxylic acid
##STR00537## 1-(6-fluorobenzo[d][1,3]dioxol-5-
yl)cyclopropanecarboxylic acid ##STR00538##
1-(2,3-dihydrobenzofuran-6- yl)cyclopropanecarboxylic acid
##STR00539## 1-(chroman-6-yl)cyclopropanecarboxylic acid
##STR00540## 1-(4-hydroxy-4-methoxychroman-6-
yl)cyclopropanecarboxylic acid ##STR00541## 1-(4-oxochroman-6-
yl)cyclopropanecarboxylic acid ##STR00542## 1-(3,4-
dichlorophenyl)cyclopropanecarboxylic acid ##STR00543##
1-(2,3-dihydronzo[b][1,4]dioxin-6- yl)cyclopropanecarboxylic acid
##STR00544## 1-(benzofuran-6- yl)cyclopropanecarboxylic acid
##STR00545##
[0350] Specific Procedures: Synthesis of Aminoindole Building
Blocks
Example 28
3-Methyl-1H-indol-6-amine
##STR00546##
[0351] (3-Nitro-phenyl)-hydrazine hydrochloride salt
[0352] 3-Nitro-phenylamine (27.6 g, 0.2 mol) was dissolved in the
mixture of H.sub.2O (40 mL) and 37% HCl (40 mL). A solution of
NaNO.sub.2 (13.8 g, 0.2 mol) in H.sub.2O (60 mL) was added to the
mixture at 0.degree. C., and then a solution of SnCl.sub.2.H.sub.2O
(135.5 g, 0.6 mol) in 37% HCl (100 mL) was added at that
temperature. After stirring at 0.degree. C. for 0.5 h, the
insoluble material was isolated by filtration and was washed with
water to give (3-nitrophenyl)hydrazine hydrochloride (27.6 g,
73%).
##STR00547##
N-(3-Nitro-phenyl)-N'-propylidenehydrazine
[0353] Sodium hydroxide solution (10%, 15 mL) was added slowly to a
stirred suspension of (3-nitrophenyl)hydrazine hydrochloride (1.89
g, 10 mmol) in ethanol (20 mL) until pH 6. Acetic acid (5 mL) was
added to the mixture followed by propionaldehyde (0.7 g, 12 mmol).
After stirring for 3 h at room temperature, the mixture was poured
into ice-water and the resulting precipitate was isolated by
filtration, washed with water and dried in air to obtain
(E)-1-(3-nitrophenyl)-2-propylidenehydrazine, which was used
directly in the next step.
##STR00548##
3-Methyl-4-nitro-1H-indole 3 and 3-methyl-6-nitro-1H-indole
[0354] A mixture of (E)-1-(3-nitrophenyl)-2-propylidenehydrazine
dissolved in 85% H.sub.3PO.sub.4 (20 mL) and toluene (20 mL) was
heated at 90-100.degree. C. for 2 h. After cooling, toluene was
removed under reduced pressure. The resultant oil was basified to
pH 8 with 10% NaOH. The aqueous layer was extracted with EtOAc (100
mL.times.3). The combined organic layers were dried, filtered and
concentrated under reduced pressure to afford the mixture of
3-methyl-4-nitro-1H-indole and 3-methyl-6-nitro-1H-indole[1.5 g in
total, 86%, two steps from (3-nitrophenyl)hydrazine hydrochloride]
which was used to the next step without further purification.
##STR00549##
3-Methyl-1H-indol-6-amine
[0355] The crude mixture from previous steps (3 g, 17 mmol) and 10%
Pd--C(0.5 g) in ethanol (30 mL) was stirred overnight under H.sub.2
(1 atm) at room temperature. Pd--C was filtered off and the
filtrate was concentrated under reduced pressure. The solid residue
was purified by column to give 3-methyl-1H-indol-6-amine (0.6 g,
24%). .sup.1H NMR (CDCl.sub.3) .delta. 7.59 (br s. 1H), 7.34 (d,
J=8.0 Hz, 1H), 6.77 (s, 1H), 6.64 (s, 1H), 6.57 (m, 11H), 3.57
(brs, 2H), 2.28 (s, 3H); MS (ESI) m/e (M+H.sup.+) 147.2.
Example 29
3-tert-Butyl-1H-indol-5-amine
##STR00550##
[0356] 3-tert-Butyl-S-nitro-1H-indole
[0357] To a mixture of 5-nitro-1H-indole (6.0 g, 37 mmol) and
AlCl.sub.3 (24 g, 0.18 mol) in CH.sub.2Cl.sub.2 (100 mL) at
0.degree. C. was added 2-bromo-2-methyl-propane (8.1 g, 37 mmol)
dropwise. After being stirred at 15.degree. C. overnight, the
mixture was poured into ice (100 mL). The precipitated salts were
removed by filtration and the aqueous layer was extracted with
CH.sub.2Cl.sub.2 (30 mL.times.3). The combined organic layers were
washed with water, brine, dried over Na.sub.2SO.sub.4 and
concentrated under vacuum to obtain the crude product, which was
purified by column chromatography on silica gel (petroleum
ether/ethyl acetate=20:1) to give 3-tert-butyl-5-nitro-1H-indole
(2.5 g, 31%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.49 (d.
J=1.6 Hz, 1H), 8.31 (brs, 1H), 8.05 (dd, J=2.0, 8.8 Hz, 1H), 7.33
(d, J=8.8 Hz, 1H), 6.42 (d, J=1.6 Hz, 1H), 1.42 (s, 9H).
##STR00551##
3-tert-Butyl-1H-indol-5-amine
[0358] To a solution of 3-tert-butyl-5-nitro-1H-indole (2.5 g, 12
mmol) in MeOH (30 mL) was added Raney Nickel (0.2 g) under N.sub.2
protection. The mixture was stirred under hydrogen atmosphere (1
atm) at 15.degree. C. for 1 h. The catalyst was filtered off and
the filtrate was concentrated to dryness under vacuum. The residue
was purified by preparative HLPC to afford
3-tert-butyl-1H-indol-5-amine (0.43 g, 19%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.72 (br.s, 1H), 7.11 (d, J=8.4 Hz,
1H), 6.86 (d, J=2.0 Hz, 1H), 6.59 (dd, J=2.0, 8.4 Hz, 1H), 6.09 (d,
J=1.6 Hz, 1H), 1.37 (s, 9H); MS (ESI) m/e (M+H.sup.+) 189.1.
Example 30
2-tert-Butyl-6-fluoro-1H-indol-5-amine and
6-tert-butoxy-2-tert-butyl-1H-indol-5-amine
##STR00552##
[0359] 2-Bromo-5-fluoro-4-nitroaniline
[0360] To a mixture of 3-fluoro-4-nitroaniline (6.5 g, 42.2 mmol)
in AcOH (80 mL) and chloroform (25 mL) was added dropwise Br.sub.2
(2.15 mL, 42.2 mmol) at 0.degree. C. After addition, the resulting
mixture was stirred at room temperature for 2 h and then poured
into ice water. The mixture was basified with aqueous NaOH (10%) to
pH.about.8.0-9.0 under cooling and then extracted with EtOAc (50
mL.times.3). The combined organic layers were washed with water (80
mL.times.2) and brine (100 mL), dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to give
2-bromo-5-fluoro-4-nitroaniline (9 g, 90%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.26 (d, J=8.0, z, 1H), 7.07 (brs, 2H), 6.62
(d, J=9.6 Hz, 1H).
##STR00553##
2-(3,3-Dimethylbut-1-ynyl)-5-fluoro-4-nitroaniline
[0361] A mixture of 2-bromo-5-fluoro-4-nitroaniline (9.0 g, 38.4
mmol), 3,3-dimethyl-but-1-yne (9.95 g, 121 mmol), CuI (0.5 g 2.6
mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (3.4 g, 4.86 mmol) and Et.sub.3N
(14 mL, 6.9 mmol) in toluene (100 mL) and water (50 mL) was heated
at 70.degree. C. for 4 h. The aqueous layer was separated and the
organic layer was washed with water (80 mL.times.2) and brine (100
mL), dried over Na.sub.2SO.sub.4 and concentrated under reduced
pressure to dryness. The residue was recrystallized with ether to
afford 2-(3,3-dimethylbut-1-ynyl)-5-fluoro-4-nitroaniline (4.2 g,
46%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 7.84 (d, J=8.4
Hz, 1H), 6.84 (brs, 2H), 6.54 (d, J=14.4 Hz, 1H), 1.29 (s, 9H).
##STR00554##
N-(2-(3,3Dimethylbut-1-ynyl)-5-fluoro-4-nitrophenyl)butyramide
[0362] To a solution of
2-(3,3-dimethylbut-1-ynyl)-5-fluoro-4-nitroaniline (4.2 g, 17.8
mmol) in dichloromethane (50 mL) and Et.sub.3N (10.3 mL, 71.2 mmol)
was added butyryl chloride (1.9 g, 17.8 mmol) at 0.degree. C. The
mixture was stirred at room temperature for 1 h and then poured
into water. The aqueous phase was separated and the organic layer
was washed with water (50 mL.times.2) and brine (100 mL), dried
over Na.sub.2SO.sub.4 and concentrated under reduced pressure to
dryness. The residue was washed with ether to give
N-(2-(3,3-dimethylbut-1-ynyl)-5-fluoro-4-nitrophenyl)butyramide
(3.5 g, 67%), which was used in the next step without further
purification.
##STR00555##
2-tert-Butyl-6-fluoro-5-nitro-1H-indole
[0363] A solution of
N-(2-(3,3-dimethylbut-1-ynyl)-5-fluoro-4-nitrophenyl)butyramide
(3.0 g, 9.8 mmol) and TBAF (4.5 g, 17.2 mmol) in DMF (25 mL) was
heated at 100.degree. C. overnight. The mixture was poured into
water and then extracted with EtOAc (80 mL.times.3). The combined
extracts were washed with water (50 mL) and brine (50 mL), dried
over Na.sub.2SO.sub.4 and concentrated under reduced pressure to
dryness. The residue was purified by column chromatography on
silica gel (petroleum ether/ethyl acetate 20:1) to give compound
2-tert-butyl-6-fluoro-5-nitro-1H-indole (1.5 g, 65%). .sup.1H-NMR
(400 MHz, CDCl.sub.3) .delta. 8.30 (d, J=7.2 Hz, 1H), 7.12 (d,
J=11.6 Hz, 1H), 6.35 (d, J=1.2 Hz, 1H), 1.40 (s, 9H).
##STR00556##
2-tert-Butyl-6-fluoro-1H-indol-5-amine
[0364] A suspension of 2-tert-butyl-6-fluoro-5-nitro-1H-indole (1.5
g, 6.36 mmol) and Ni (0.5 g) in MeOH (20 mL) was stirred under
H.sub.2 atmosphere (1 atm) at the room temperature for 3 h. The
catalyst was filtered off and the filtrate was concentrated under
reduced pressure to dryness. The residue was recrystallized in
ether to give 2-tert-butyl-6-fluoro-1H-indol-5-amine (520 mg, 38%).
.sup.1H-NMR (300 MHz, DMSO-d.sub.6) .delta. 10.46 (brs, 1H); 6.90
(d, J=8.7 Hz, 1H), 6.75 (d, J=9.0 Hz, 1H), 5.86 (s, 1H), 4.37 (brs,
2H), 1.29 (s, 9H); MS (ESI) m/e 206.6.
##STR00557##
6-tert-Butoxy-2-tert-butyl-5-nitro-1H-indole
[0365] A solution of
N-(2-(3,3-dimethylbut-1-ynyl)-5-fluoro-4-nitrophenyl)butyramide
(500 mg, 1.63 mmol) and t-BuOK (0.37 g, 3.26 mmol) in DMF (10 mL)
was heated at 70.degree. C. for 2 h. The mixture was poured into
water and then extracted with EtOAc (50 mL.times.3). The combined
extracts were washed with water (50 mL) and brine (50 mL), dried
over Na.sub.2SO.sub.4 and concentrated under reduced pressure to
give 6-tert-butoxy-2-tert-butyl-5-nitro-1H-indole (100 mg, 21%).
.sup.1H-NMR (300 MHz, DMSO-d.sub.6) .delta. 11.35 (brs, 1H), 7.99
(s, 1H), 7.08 (s, 1H), 6.25 (s, 11H), 1.34 (s, 9H), 1.30 (s,
9H).
##STR00558##
6-tert-Butoxy-2-tert-butyl-1H-indol-5-amine
[0366] A suspension of 6-tert-butoxy-2-tert-butyl-5-nitro-1H-indole
(100 mg, 0.36 mmol) and Raney Ni (0.5 g) in MeOH (15 mL) was
stirred under H.sub.2 atmosphere (1 atm) at the room temperature
for 2.5 h. The catalyst was filtered off and the filtrate was
concentrated under reduced pressure to dryness. The residue was
recrystallized in ether to give
6-tert-butoxy-2-tert-butyl-1H-indol-5-amine (30 mg, 32%).
.sup.1H-NMR (300 MHz, MeOD) 6.98 (s, 1H), 6.90 (s, 1H), 5.94 (d,
J.degree. 0.6 Hz, 1H), 1.42 (s, 9H), 1.36 (s, 9H); MS (ESI) m/e
205.0.
Example 31
1-tert-Butyl-1H-indol-5-amine
##STR00559##
[0367] N-tert-Butyl-4-nitroaniline
[0368] A solution of 1-fluoro-4-nitro-benzene (1 g, 7.1 mmol) and
tert-butylamine (1.5 g, 21 mmol) in DMSO (5 mL) was stirred at
75.degree. C. overnight. The mixture was poured into water (10 mL)
and extracted with EtOAc (7 mL.times.3). The combined organic
layers were washed with water, brine, dried over Na.sub.2SO.sub.4
and concentrated under vacuum to dryness. The residue was purified
by column chromatography on silica gel (petroleum ether/ethyl
acetate 30:1) to afford N-tert-butyl-4-nitroaniline (1 g, 73%). 1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.03-8.00 (m, 2H), 6.61-6.57 (m,
2H), 4.67 (brs, 1H), 1.42 (s, 9H).
##STR00560##
(2-Bromo-4-nitro-phenyl)-tert-butyl-amine
[0369] To a solution of N-tert-butyl-4-nitroaniline (1 g, 5.1 mmol)
in AcOH (5 mL) was added Br.sub.2 (0.86 g, 54 mmol) dropwise at
15.degree. C. After addition, the mixture was stirred at 30.degree.
C. for 30 min and then filtered. The filter cake was basified to pH
8-9 with aqueous NaHCO.sub.3. The aqueous layer was extracted with
EtOAc (10 mL.times.3). The combined organic layers were washed with
water, brine, dried over Na.sub.2SO.sub.4 and concentrated under
vacuum to give (2-bromo-4-nitro-phenyl)-tert-butyl-amine (0.6 g,
43%). .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 8.37 (dd, J=2.4 Hz,
1H), 8.07 (dd, J=2.4, 9.2 Hz, 1H), 6.86 (d, J=9.2 Hz, 1H), 5.19
(brs, 1H), 1.48 (s, 9H).
##STR00561##
tert-Butyl-(4-nitro-2-trimethylsilanylethynyl-phenyl)-amine
[0370] To a solution of (2-bromo-4-nitro-phenyl)-tert-butyl-amine
(0.6 g, 2.2 mmol) in Et.sub.3N (10 mL) was added
Pd(PPh.sub.3).sub.2Cl.sub.2 (70 mg, 0.1 mmol), CuI (20.9 mg, 0.1
mmol) and ethynyl-trimethyl-silane (0.32 g, 3.3 mmol) successively
under Nz protection. The reaction mixture was heated at 70.degree.
C. overnight. The solvent was removed under vacuum and the residue
was washed with EtOAc (10 mL.times.3). The combined organic layers
were washed with water, brine, dried over Na.sub.2SO.sub.4 and
concentrated under vacuum to dryness. The residue was purified by
column chromatography on silica gel (petroleum ether/ethyl acetate
20:1) to afford
tert-butyl-(4-nitro-2-trimethylsilanylethynyl-phenyl)-amine (100
mg, 16%). .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 8.20 (d, J=2.4,
Hz, 1H), 8.04 (dd, J=2.4, 9.2 Hz, 1H), 6.79 (d, J=9.6 Hz, 1H), 5.62
(brs, 1H), 1.41 (s, 9H), 0.28 (s, 9H).
##STR00562##
1-tert-Butyl-5-nitro-1H-indole
[0371] To a solution of
tert-butyl-(4-nitro-2-trimethylsilanylethynyl-phenyl)-amine (10 mg,
0.035 mmol) in DMF (2 mL), was added CuI (13 mg, 0.07 mmol) under
N.sub.2 protection. The reaction mixture was stirred at 100.degree.
C. overnight. At this time, EtOAc (4 mL) was added to the mixture.
The mixture was filtered and the filtrate was washed with water,
brine, dried over Na.sub.2SO.sub.4 and concentrated under vacuum to
obtain 1-tert-butyl-5-nitro-1H-indole (7 mg, 93%). .sup.1H-NMR
(CDCl.sub.3, 300 MHz) .delta. 8.57 (d, J=2.1 Hz, 1H), 8.06 (dd,
J=2.4, 9.3 Hz, 1H), 7.65 (d, J=9.3 Hz, 1H), 7.43 (d, J=3.3 Hz, 1H),
6.63 (d, J=3.3 Hz, 1H), 1.76 (s, 9H).
##STR00563##
1-ter-Butyl-1H-indol-5-amine
[0372] To a solution of 1-tert-butyl-S-nitro-1H-indole (6.5 g,
0.030 mol) in MeOH (100 mL) was added Raney Nickel (0.65 g, 10%)
under N.sub.2 protection. The mixture was stirred under hydrogen
atmosphere (1 atm) at 30.degree. C. for 1 h. The catalyst was
filtered off and the filtrate was concentrated under vacuum to
dryness. The residue was purified by column chromatography on
silica gel (PE/EtOAc 1:2) to give 1-tert-butyl-1H-indol-5-amine
(2.5 g, 45%). .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 7.44 (d,
J=8.8 Hz, 1H), 7.19 (dd, J=3.2 Hz, 1H), 6.96 (d, J=2.0 Hz, 1H),
6.66 (d, J=2.0, 8.8 Hz, 1H), 6.26 (d, J=3.2 Hz, 1H), 1.67 (s, 9H).
MS (ESI) m/e (M+H.sup.+) 189.2.
Example 32
2-tert-Butyl-1-methyl-1H-indol-5-amine
##STR00564##
[0373] (2-Bromo-4-nitro-phenyl)-methyl-amine
[0374] To a solution of methyl-(4-nitro-phenyl)-amine (15.2 g, 0.1
mol) in AcOH (150 mL) and CHCl.sub.3 (50 mL) was added Br.sub.2
(16.0 g, 0.1 mol) dropwise at 5.degree. C. The mixture was stirred
at 10.degree. C. for 1 h and then basified with sat. aq.
NaHCO.sub.3. The resulting mixture was extracted with EtOAc (100
mL.times.3), and the combined organics were dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under vacuum to give
(2-bromo-4-nitro-phenyl)-methyl-amine
(2-bromo-4-nitro-phenyl)-methyl-amine (23.0 g, 99%), which was used
in the next step without further purification. .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 8.37 (d, J=2.4 Hz, 1H), 8.13 (dd, J=2.4,
9.0 Hz, 1H), 6.58 (d, J=9.0 Hz, l H), 5.17 (brs, 1H), 3.01 (d,
J=5.4 Hz, 3H).
##STR00565##
[2-(3,3-Dimethyl-but-1-ynyl)-4-nitro-phenyl]-methyl-amine
[0375] To a solution of (2-bromo-4-nitro-phenyl)-methyl-amine (22.5
g, 97.4 mmol) in toluene (200 mL) and water (100 mL) were added
Et.sub.3N (19.7 g, 195 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (6.8 g,
9.7 mmol), CuI (0.7 g, 3.9 mmol) and 3,3-dimethyl-but-1-yne (16.0
g, 195 mmol) successively under N.sub.2 protection. The mixture was
heated at 70.degree. C. for 3 hours and then cooled down to room
temperature. The resulting mixture was extracted with EtOAc (100
mL.times.3). The combined organic extracts were dried over
anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum to give
[2-(3,3-dimethyl-but-1-ynyl)-4-nitro-phenyl]-methyl-amine (20.1 g,
94%), which was used in the next step without further purification.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.15 (d, J=2.4 Hz, 1H),
8.08 (dd, J=2.8, 9.2 Hz, 1H), 6.50 (d, J=92 Hz, 1H), 5.30 (brs,
1H), 3.00 (s, 3H), 1.35 (s, 9H).
##STR00566##
2-tert-Butyl-1-methyl-5-nitro-1H-indole
[0376] A solution of
[2-(3,3-dimethyl-but-1-ynyl)-4-nitro-phenyl]-methyl-amine (5.0 g,
22.9 mmol) and TBAF (23.9 g, 91.6 mmol) in THF (50 mL) was heated
at reflux overnight. The solvent was removed by evaporation under
vacuum and the residue was dissolved in brine (100 mL) and EtOAc
(100 mL). The organic phase was separated, dried over
Na.sub.2SO.sub.4 and evaporated under vacuum to give
2-tert-butyl-1-methyl-5-nitro-1H-indole (5.0 g, 99%), which was
used in the next step without further purification. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.47 (d, J=2.4 Hz, 1H), 8.07 (dd,
J=2.4, 9.2 Hz, 1H), 7.26-7.28 (m, 1H), 6.47 (s, 1H), 3.94 (s, 3H),
1.50 (s, 9H).
##STR00567##
2-tert-Butyl-1-methyl-1H-indol-5-amine
[0377] To a solution of 2-tert-butyl-1-methyl-5-nitro-1H-indole
(3.00 g, 13.7 mmol) in MeOH (30 mL) was added Raney Ni (0.3 g)
under nitrogen atmosphere. The mixture was stirred under hydrogen
atmosphere (1 atm) at room temperature overnight. The mixture was
filtered through a Celite pad and the filtrate was evaporated under
vacuum. The crude residue was purified by column chromatography on
silica gel (P.E/EtOAc 20:1) to give
2-tert-butyl-1-methyl-1H-indol-5-amine (1.7 g, 66%). .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 7.09 (d, J=8.4 Hz, 1H), 6.89-6.9 (m,
1H), 6.66 (dd, J=2.4, 8.7 Hz, 1H), 6.14 (d, J=0.6 Hz, 1H), 3.83 (s,
3H), 3.40 (brs, 2H), 1.45 (s, 9H); MS (ESI) m/e (M+H.sup.+)
203.1.
Example 33
2-Cyclopropyl-1H-indol-5-amine
##STR00568##
[0378] 2-Bromo-4-nitroaniline
[0379] To a solution of 4-nitro-aniline (25 g, 0.18 mol) in HOAc
(150 mL) was added liquid Br.sub.2 (30 g, 0.19 mol) dropwise at
room temperature. The mixture was stirred for 2 hours. The solid
was collected by filtration and poured into water (100 mL), which
was basified with sat. aq. NaHCO.sub.3 to pH 7 and extracted with
EtOAc (300 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4 and evaporated under reduced pressure to
give 2-bromo-4-nitroaniline (30 g, 80%), which was directly used in
the next step.
##STR00569##
2-(Cyclopropylethynyl)-4-nitroaniline
[0380] To a deoxygenated solution of 2-bromo-4-nitroaniline (2.17
g, 0.01 mmol), ethynyl-cyclopropane (1 g, 15 mmol) and CuI (10 mg,
0.05 mmol) in triethylamine (20 mL) was added
Pd(PPh.sub.3).sub.2Cl.sub.2 (210 mg, 0.3 mmol) under N.sub.2. The
mixture was heated at 70.degree. C. and stirred for 24 hours. The
solid was filtered off and washed with EtOAc (50 mL.times.3). The
filtrate was evaporated under reduced pressure, and the residue was
purified by column chromatography on silica gel (petroleum
ether/ethyl acetate=10/1) to give
2-(cyclopropylethynyl)-4-nitroaniline (470 mg, 23%). .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 8.14 (d, J=2.7 Hz, 1H), 7.97 (dd,
J=2.7, 9.0 Hz, 1H), 6.63 (d, J=9.0 Hz, 1H), 4.81 (brs, 2H),
1.55-1.46 (m, 1H), 0.98-0.90 (m, 2H), 0.89-0.84 (m, 2H).
##STR00570##
N-(2-(Cyclopropylethynyl)phenyl)-4-nitrobutyramide
[0381] To a solution of 2-(cyclopropylethynyl)-4-nitroaniline (3.2
g, 15.8 mmol) and pyridine (2.47 g, 31.7 mmol) in CH.sub.2Cl.sub.2
(60 mL) was added butyryl chloride (2.54 g, 23.8 mmol) at 0.degree.
C. The mixture was warmed to room temperature and stirred for 3
hours. The resulting mixture was poured into ice-water. The organic
layer was separated. The aqueous phase was extracted with
CH.sub.2Cl.sub.2 (30 mL.times.3). The combined organic layers were
dried over anhydrous Na.sub.2SO.sub.4 and evaporated under reduced
pressure to give the crude product, which was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate=10/1)
to give N-(2-(cyclopropylethynyl)phenyl)-4-nitrobutyramide (3.3 g,
76%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.61 (d, J=9.2 Hz,
1H), 8.22 (d, J=2.8 Hz, 1H), 8.18 (brs, 1H), 8.13 (dd, J 2.4, 9.2
Hz, 1H), 2.46 (t, J=7.2 Hz, 2H), 1.83-1.76 (m, 2H), 1.59-1.53 (m,
1H), 1.06 (t, J=7.2 Hz, 3H), 1.03-1.01 (m, 2H), 0.91-0.87 (m,
2H).
##STR00571##
2-Cyclopropyl-5-nitro-1H-indole
[0382] A mixture of
N-(2-(cyclopropylethynyl)phenyl)-4-nitrobutyramide (3.3 g, 0.01
mol) and TBAF (9.5 g, 0.04 mol) in THF (100 mL) was heated at
reflux for 24 hours. The mixture was cooled to the room temperature
and poured into ice water. The mixture was extracted with
CH.sub.2Cl.sub.2 (50 mL.times.3). The combined organic layers were
dried over anhydrous Na.sub.2SO.sub.4 and evaporated under reduced
pressure. The residue was purified by column chromatography on
silica gel (petroleum ether/ethyl acetate=10/1) to give
2-cyclopropyl-5-nitro-1H-indole (1.3 g, 64%). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.44 (d, J=2.0 Hz, 1H), 8.40 (brs, 1H), 8.03
(dd, J=2.0, 8.8 Hz, 1H), 7.30 (d, J=8.8 Hz, 1N), 6.29 (d, J=0.8 Hz,
1H), 2.02-1.96 (m, 1H) 1.07-1.02 (m, 2H), 0.85-0.81 (m, 2H).
##STR00572##
2-Cyclopropyl-1H-indol-5-amine
[0383] To a solution of 2-cyclopropyl-5-nitro-1H-indole (1.3 g, 6.4
mmol) in MeOH (30 mL) was added Raney Nickel (0.3 g) under nitrogen
atmosphere. The mixture was stirred under hydrogen atmosphere (1
atm) at room temperature overnight. The catalyst was filtered
through a Celite pad and the filtrate was evaporated under vacuum
to give the crude product, which was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate=5/1) to
give 2-cyclopropyl-1H-indol-5-amine (510 mg, 56%). .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 6.89 (d, J=8.4 Hz, 1H), 6.50 (d, J=1.6 Hz,
1H), 6.33 (dd, J=2.0, 8.4 Hz, 1H), 5.76 (s, 1H), 4.33 brs, 2H),
1.91-1.87 (m, 1H), 0.90-0.85 (m, 2H), 0.70-0.66 (m, 2H); MS (ESI)
m/e (M+H.sup.+) 173.2.
Example 34
3-tert-Butyl-1H-indol-5-amine
##STR00573##
[0384] 3-tert-Butyl-5-nitro-1H-indole
[0385] To a mixture of 5-nitro-1H-indole (6 g, 36.8 mmol) and
AlCl.sub.3 (24 g, 0.18 mol) in CH.sub.2Cl.sub.2 (100 mL) was added
2-bromo-2-methyl-propane (8.1 g, 36.8 mmol) dropwise at 0.degree.
C. After being stirred at 15.degree. C. overnight, the reaction
mixture was poured into ice (100 mL). The precipitated salts were
removed by filtration and the aqueous layer was extracted with
CH.sub.2Cl.sub.2 (30 mL.times.3). The combined organic layers were
washed with water, brine, dried over Na.sub.2SO.sub.4 and
concentrated under vacuum to obtain the crude product, which was
purified by column chromatography on silica gel (petroleum
ether/ethyl acetate 20:1) to give 3-tert-butyl-5-nitro-1H-indole
(2.5 g, 31%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.49 (d,
J=1.6 Hz, 1H), 8.31 (brs, 1H), 8.05 (dd, J=2.0, 8.8 Hz, 1H), 7.33
(d, J=8.8 Hz, 1H), 6.42 (d, J=1.6 Hz, 1H), 1.42 (s, 9H).
##STR00574##
3-tert-Butyl-1H-indol-5-amine
[0386] To a solution of 3-tert-butyl-5-nitro-1H-indole (2.5 g, 11.6
mmol) in MeOH (30 mL) was added Raney Nickel (0.2 g) under N.sub.2
protection. The mixture was stirred under hydrogen atmosphere (1
atm) at 15.degree. C. for 1 hr. The catalyst was filtered off and
the filtrate was concentrated under vacuum to dryness. The residue
was purified by preparative HLPC to afford
3-tert-butyl-1H-indol-5-amine (0.43 g, 19%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.72 (brs, 1H), 7.11 (d, J=8.4 Hz,
1H), 6.86 (d, J=2.0 Hz, 1H), 6.59 (dd, J=2.0, 8.4 Hz, 1H), 6.09 (d,
J=1.6 Hz, 1H), 1.37 (s, 9H); MS (ESI) m/e (M+H.sup.+) 189.1.
Example 35
2-Phenyl-1H-indol-5-amine
##STR00575##
[0387] 2-Bromo-4-nitroaniline
[0388] To a solution of 4-nitroaniline (50 g, 0.36 mol) in AcOH
(500 mL) was added liquid Br.sub.2 (60 g, 0.38 mol) dropwise at
5.degree. C. The mixture was stirred for 30 min at that
temperature. The insoluble solid was collected by filtration and
poured into EtOAc (200 mL). The mixture was basified with saturated
aqueous NaHCO.sub.3 to pH 7. The organic layer was separated. The
aqueous phase was extracted with EtOAc (300 mL.times.3). The
combined organic layers were dried and evaporated under reduced
pressure to give 2-bromo-4-nitroaniline (56 g, 72%), which was
directly used in the next step.
##STR00576##
4-Nitro-2-(phenylethynyl)aniline
[0389] To a deoxygenated solution of 2-bromo-4-nitroaniline (2.17
g, 0.01 mmol), ethynyl-benzene (1.53 g, 0.015 mol) and CuI (10 mg,
0.05 mmol) in triethylamine (20 mL) was added
Pd(PPh.sub.3).sub.2Cl.sub.2 (210 mg, 0.2 mmol) under N.sub.2. The
mixture was heated at 70.degree. C. and stirred for 24 hours. The
solid was filtered off and washed with EtOAc (50 mL.times.3). The
filtrate was evaporated under reduced pressure and the residue was
purified by column chromatography on silica gel (petroleum
ether/ethyl acetate=10/1) to give 4-nitro-2-(phenylethynyl)aniline
(340 mg, 14%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.37-8.29
(m, 1H), 8.08-8.00 (m, 1H), 7.56-7.51 (m, 2H), 7.41-7.37 (m, 3H),
6.72 (m, 1H), 4.95 (brs, 2H).
##STR00577##
N-(2-(Phenylethynyl)phenyl)-4-nitrobutyramide
[0390] To a solution of 4-nitro-2-(phenylethynyl)aniline (17 g,
0.07 mmol) and pyridine (11.1 g, 0.14 mol) in CH.sub.2Cl.sub.2 (100
mL) was added butyryl chloride (11.5 g, 0.1 mol) at 0.degree. C.
The mixture was warmed to room temperature and stirred for 3 hours.
The resulting mixture was poured into ice-water. The organic layer
was separated. The aqueous phase was extracted with
CH.sub.2Cl.sub.2 (30 mL.times.3). The combined organic layers were
dried over anhydrous Na.sub.2SO.sub.4 and evaporated under reduced
pressure. The residue was purified by column chromatography on
silica gel (petroleum ether/ethyl acetate=10/1) to give
N-(2-(phenylethynyl)phenyl)-4-nitrobutyramide (12 g, 55%). .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 8.69 (d, J=9.2 Hz, 1H), 8.39 (d,
J=2.8 Hz, 1H), 8.25-8.20 (m, 2H), 7.58-7.55 (m, 2H), 7.45-7.42 (m,
3H), 2.49 (t, J=7.2 Hz, 2H), 1.85-1.79 (m, 2H), 1.06 (t, J=7.2 Hz,
3H).
##STR00578##
5-Nitro-2-phenyl-1H-indole
[0391] A mixture of N-(2-(phenylethynyl)phenyl)-4-nitrobutyramide
(5.0 g, 0.020 mol) and TBAF (12.7 g, 0.050 mol) in THF (30 mL) was
heated at reflux for 24 h. The mixture was cooled to room
temperature and poured into ice water. The mixture was extracted
with CH.sub.2Cl.sub.2 (50 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4 and evaporated under
reduced pressure. The residue was purified by column chromatography
on silica gel (petroleum ether/ethyl acetate=10/1) to give
5-nitro-2-phenyl-1H-indole (3.3 g, 69%). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.67 (s, 1H), 8.06 (dd, J=2.0, 8.8 Hz, 1H),
7.75 (d, J=7.6 Hz, 2H), 7.54 (d, J=8.8 Hz, 1H), 7.45 (t, J=7.6 Hz,
2H), 7.36 (t, J=7.6 Hz, 1H). 6.95 (s, 1H).
##STR00579##
2-Phenyl-1H-indol-5-amine
[0392] To a solution of 5-nitro-2-phenyl-1H-indole (2.83 g, 0.01
mol) in MeOH (30 mL) was added Raney Ni (510 mg) under nitrogen
atmosphere. The mixture was stirred under hydrogen atmosphere (1
atm) at room temperature overnight. The catalyst was filtered
through a Celite pad and the filtrate was evaporated under vacuum
to give the crude product, which was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate=5/1) to
give 2-phenyl-1H-indol-5-amine (1.6 g, 77%). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.76 (d, J=7.6 Hz, 2H), 7.39 (t, J=7.6 Hz, 2H),
7.24 (t, J=7.6 Hz, 1H), 7.07 (d, J=8.4 Hz, 1H), 6.64 (d, J=1.6 Hz,
1H), 6.60 (d, J=1.2 Hz, 1H), 6.48 (dd, J=2.0, 8.4 Hz, 1H), 4.48
(brs, 2H); MS (ESI) m/e (M+H.sup.+) 209.0.
Example 36
2-tert-Butyl-4-fluoro-1H-indol-5-amine
##STR00580##
[0393] 2-Bromo-3-fluoroaniline
[0394] To a solution of 2-bromo-1-fluoro-3-nitrobenzene (1.0 g, 5.0
mmol) in CH.sub.3OH (50 mL) was added NiCl.sub.2 (2.2 g 10 mmol)
and NaBH.sub.4 (0.50 g 14 mmol) at 0.degree. C. After the addition,
the mixture was stirred for 5 min. Water (20 mL) was added and the
mixture was extracted with EtOAc (20 mL.times.3). The organic
layers were dried over anhydrous Na.sub.2SO.sub.4 and evaporated
under vacuum to give 2-bromo-3-fluoroaniline (600 mg, 70%). .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.07-7.02 (m, 1H), 6.55-6.49 (m,
1H), 4.22 (br s, 2H).
##STR00581##
N-(2-Bromo-3-fluorophenyl)butyramide
[0395] To a solution of 2-bromo-3-fluoroaniline (2.0 g, 11 mmol) in
CH.sub.2Cl.sub.2 (50 mL) was added butyryl chloride (1.3 g, 13
mmol) and pyridine (1.7 g, 21 mmol) at 0.degree. C. The mixture was
stirred at room temperature for 24 h. Water (20 mL) was added and
the mixture was extracted with CH.sub.2Cl.sub.2 (50 mL.times.3).
The organic layers were dried anhydrous over Na.sub.2SO.sub.4 and
evaporated under vacuum to give
N-(2-bromo-3-fluorophenyl)butyramide (2.0 g, 73%), which was
directly used in the next step.
##STR00582##
N-(2-(3,3-Dimethylbut-1-ynyl)-3-fluorophenyl)butyramide
[0396] To a solution of N-(2-bromo-3-fluorophenyl)butyramide (2.0
g, 7.0 mmol) in Et.sub.3N (100 mL) was added 4,4-dimethylpent-2-yne
(6.0 g, 60 mmol), Cu (70 mg, 3.8 mmol), and
Pd(PPh.sub.3).sub.2Cl.sub.2 (500 mg) successively at room
temperature under N.sub.2. The mixture was heated at 80.degree. C.
overnight. The cooled mixture was filtered and the filtrate was
extracted with EtOAc (40 mL.times.3). The organic layers were
washed with sat. NaCl, dried over anhydrous Na.sub.2SO.sub.4, and
evaporated under vacuum. The crude compound was purified by column
chromatography on silica gel (10% EtOAc in petroleum ether) to give
N-(2-(3,3-dimethylbut-1-ynyl)-3-fluorophenyl)butyramide (1.1 g,
55%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.20 (d, J=7.6,
1H), 7.95 (s, 1H), 7.21 (m, 1H), 6.77 (t, J=7.6 Hz, 1H), 2.39 (t,
J=7.6 Hz, 2H), 1.82-1.75 (m, 2H), 1.40 (s, 9H), 1.12 (t, J=7.2 Hz,
3H).
##STR00583##
2-tert-Butyl-4-fluoro-1H-indole
[0397] To a solution of
N-(2-(3,3-dimethylbut-1-ynyl)-3-fluorophenyl)butyramide (6.0 g, 20
mmol) in DMF (100 mL) was added t-BuOK (5.0 g, 50 mmol) at room
temperature. The mixture was heated at 90.degree. C. overnight
before it was poured into water and extracted with EtOAc (100
mL.times.3). The organic layers were washed with sat. NaCl and
water, dried over anhydrous Na.sub.2SO.sub.4, and evaporated under
vacuum to give 2-tert-butyl-4-fluoro-1H-indole (5.8 g, 97%).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.17 (br s, 1H), 7.11 (d,
J=7.2 Hz, 1H), 7.05-6.99 (m, 1H), 6.76-6.71 (m, 1H), 6.34 (m, 1H),
1.41 (s, 9H).
##STR00584##
2-tert-Butyl-4-fluoro-5-nitro-1H-indole
[0398] To a solution of 2-tert-butyl-4-fluoro-1H-indole (2.5 g, 10
mmol) in H.sub.2SO.sub.4 (30 mL) was added KNO.sub.3 (1.3 g, 10
mmol) at 0.degree. C. The mixture was stirred for 0.5 h at
-10.degree. C. The mixture was poured into water and extracted with
EtOAc (100 mL.times.3). The organic layers were washed with sat.
NaCl and water, dried over anhydrous Na.sub.2SO.sub.4, and
evaporated under vacuum. The crude compound was purified by column
chromatography on silica gel (10% EtOAc in petroleum ether) to give
2-tert-butyl-4-fluoro-5-nitro-1H-indole (900 mg, 73%). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.50 (br s, 1H), 7.86 (dd, J=7.6, 8.8
Hz, 1H), 7.13 (d, J=8.8 Hz, 1H), 6.52 (dd, J=0.4, 2.0 Hz, 1H), 1.40
(s, 9H).
##STR00585##
2-tert-Butyl-4-fluoro-1H-indol-5-amine
[0399] To a solution of 2-tert-butyl-4-fluoro-5-nitro-1H-indole
(2.1 g, 9.0 mmol) in methanol (50 mL) was added NiCl.sub.2 (4.2 g,
18 mmol) and NaBH.sub.4 (1.0 g, 27 mmol) at 0.degree. C. After the
addition, the mixture was stirred for 5 min. Water (20 mL) was
added and the mixture was extracted with EtOAc (30 mL.times.3). The
organic layers were washed with sat. NaCl and water, dried over
anhydrous Na.sub.2SO.sub.4, evaporated under vacuum to give
2-tert-butyl-4-fluoro-1H-indol-5-amine (900 mg, 50%). .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 7.80 (brs, 1H), 6.91 (d, J=8.4 Hz,
1H), 6.64 (dd, J=0.9, 2.4 Hz, 1H), 6.23 (s, 1H), 1.38 (s, 9H).
Example 37
2,3,4,9-Tetrahydro-1H-carbazol-6-amine
##STR00586##
[0400] 2,3,4,9-Tetrahydro-1H-carbazol-6-amine
[0401] 6-Nitro-2,3,4,9-tetrahydro-1H-carbazole (0.100 g, 0.462
mmol) was dissolved in a 40 mL scintillation vial containing a
magnetic stir bar and 2 mL of ethanol. Tin(II) chloride dihydrate
(1.04 g, 4.62 mmol) was added to the reaction mixture and the
resulting suspension was heated at 70.degree. C. for 16 h. The
crude reaction mixture was then diluted with 15 mL of a saturated
aqueous solution of sodium bicarbonate and extracted three times
with an equivalent volume of ethyl acetate. The ethyl acetate
extracts were combined, dried over sodium sulfate, and evaporated
to dryness to yield 2,3,4,9-tetrahydro-1H-carbazol-6-amine (82 mg,
95%) which was used without further purification.
Example 38
2-tert-Butyl-7-fluoro-1H-indol-5-amine
##STR00587##
[0402] 2-Bromo-6-fluoro-4-nitro-phenylamine
[0403] To a solution of 2-fluoro-4-nitro-phenylamine (12 g, 77
mmol) in AcOH (50 mL) was added Br.sub.2 (3.9 mL, 77 mmol) dropwise
at 0.degree. C. The mixture was stirred at 20.degree. C. for 3 h.
The reaction mixture was basified with sat. aq. NaHCO.sub.3, and
extracted with EtOAc (100 mL.times.3). The combined organics were
dried over anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum
to give 2-bromo-6-fluoro-4-nitro-phenylamine (18 g, 97%). .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 8.22 (m, 1H), 7.90 (dd, J=2.4,
10.8 Hz, 1H), 4.88 (brs, 2H).
##STR00588##
2-(3,3-Dimethyl-but-1-ynyl)-6-fluoro-4-nitro-phenylamine
[0404] To a solution of 2-bromo-6-fluoro-4-nitro-phenylamine (11 g,
47 mmol) in dry Et.sub.3N (100 mL) was added CuI (445 mg, 5% mol),
Pd(PPh.sub.3).sub.2Cl.sub.2 (550 mg, 5% mol) and
3,3-dimethyl-but-1-yne (9.6 g, 120 mmol) under N.sub.2 protection.
The mixture was stirred at 80.degree. C. for 10 h. The reaction
mixture was filtered, poured into ice (100 g), and extracted with
EtOAc (50 mL.times.3). The combined organic extracts were dried
over anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum to give
the crude product, which was purified by column chromatography on
silica gel (petroleum ether/ethyl acetate 50:1) to give
2-(3,3-dimethyl-but-1-ynyl)-6-fluoro-4-nitro-phenylamine (4.0 g,
36%). .sup.1H NMR (400 MHz; CDCl.sub.3) .delta. 8.02 (d, J=1.2 Hz,
1H), 7.84 (dd, J=2.4, 10.8 Hz, 1H), 4.85 (brs, 2H), 1.36 (s,
9H).
##STR00589##
N-[2-(3,3-Dimethyl-but-1-ynyl)-6-fluoro-4-nitro-phenyl]-butyramide
[0405] To a solution of
2-(3,3-dimethyl-but-1-ynyl)-6-fluoro-4-nitro-phenylamine (4.0 g, 17
mmol) and pyridine (2.7 g, 34 mmol) in anhydrous CH.sub.2Cl.sub.2
(30 mL) was added and butyryl chloride (1.8 g, 17 mmol) dropwise at
0.degree. C. After stirring for 5 h at 0.degree. C., the reaction
mixture was poured into ice (50 g) and extracted with
CH.sub.2Cl.sub.2 (30 mL.times.3). The combined organic extracts
were dried over anhydrous Na.sub.2SO.sub.4 and evaporated under
vacuum to give
N-[2-(3,3-dimethyl-but-1-ynyl)-6-fluoro-4-nitro-phenyl)-butyramide
(3.2 g, 62%), which was used in the next step without further
purification. .sup.1H NMR (300 MHz, DMSO) .delta. 8.10 (dd, J=1.5,
2.7 Hz, 1H), 7.95 (dd, J=2.4, 9.6 Hz, 1H), 7.22 (brs, 1H), 2.45 (t,
J=7.5 Hz, 2H), 1.82 (m, 2H), 1.36 (s, 9H), 1.06 (t, J=7.5 Hz,
3H).
##STR00590##
2-tert-Butyl-7-fluoro-5-nitro-1H-indole
[0406] To a solution of
N-[2-(3,3-dimethyl-but-1-ynyl)-6-fluoro-4-nitro-phenyl]-butyramide
(3.2 g, 10 mmol) in DMF (20 mL) was added t-BuOK (2.3 g, 21 mmol)
at room temperature. The mixture was heated at 120.degree. C. for 2
g before being cooled down to room temperature. Water (50 mL) was
added to the reaction mixture and the resulting mixture was
extracted with CH.sub.2Cl.sub.2 (30 mL.times.3). The combined
organic extracts were dried over anhydrous Na.sub.2SO.sub.4 and
evaporated under vacuum to give
2-tert-butyl-7-fluoro-5-nitro-1H-indole (2.0 g, 81%), which was
used in the next step without further purification. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 9.95 (brs, 1H), 8.30 (d, J=2.1 Hz,
1H), 7.74 (dd, J=1.8, 11.1 Hz, 1H), 6.43 (dd, J=2.4, 3.3 Hz, 1H),
1.43 (s, 9H).
##STR00591##
2-tert-Butyl-7-fluoro-1H-indol-5-amine
[0407] To a solution of 2-tert-butyl-7-fluoro-5-nitro-1H-indole
(2.0 g, 8.5 mmol) in MeOH (20 mL) was added Ni (0.3 g) under
nitrogen atmosphere. The reaction mixture was stirred under
hydrogen atmosphere (1 atm) at room temperature overnight. The
catalyst was filtered off through the celite pad and the filtrate
was evaporated under vacuum. The crude product was purified by
column chromatography on silica gel (petroleum ether/ethyl acetate
100:1) to give 2-tert-butyl-7-fluoro-1H-indol-5-amine (550 mg,
24%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.87 (brs, 1H);
6.64 (d, J=1.5 Hz, 1H), 6.37 (dd, J=1.8, 12.3 Hz, i H), 6.11 (dd,
J=2.4, 3.6 Hz, 1H), 1.39 (s, 9H). MS (ESI) m/z (M+H.sup.+) 207.
Example 39
5-Amino-2-tert-butyl-1H-indole-7-carbonitrile
##STR00592##
[0408] 2-Amino-3-(3,3-dimethylbut-1-ynyl)-5-nitrobenzonitrile
[0409] To a stirred solution of 2-amino-3-bromo-5-nitrobenzonitrile
(2.4 g, 10 mmol) in dry Et.sub.3N (60 mL) was added CuI (380 mg, 5%
mol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (470 mg, 5% mol) at room
temperature. 3,3-dimethyl-but-1-yne (2.1 g, 25 mmol) was added
dropwise to the mixture at room temperature. The reaction mixture
was stirred at 80.degree. C. for 10 h. The reaction mixture was
filtered and the filtrate was poured into ice (60 g), extracted
with ethyl acetate. The phases were separated and the organic phase
was dried over Na.sub.2SO.sub.4. The solvent was removed under
vacuum to obtain the crude product, which was purified by column
chromatography (2-10% EtOAc in petroleum ether) to obtain
2-amino-3-(3,3-dimethylbut-1-ynyl)-5-nitrobenzonitrile (1.7 g,
71%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.28 (d, J=2.7 Hz,
1H), 8.27 (d, J=2.7 Hz, 1H), 5.56 (br s, 2H), 1.37 (s, 9H).
##STR00593##
2-tert-Butyl-5-nitro-1H-indole-7-carbonitrile
[0410] To a solution of
2-amino-3-(3,3-dimethylbut-1-ynyl)-5-nitrobenzonitrile (1.7 g, 7.0
mmol) in THF (35 mL) was added TBAF (9.5 g, 28 mmol) at room
temperature. The mixture was heated at reflux overnight. The
reaction mixture was cooled and the THF was removed under reduced
pressure. Water (50 ml) was added to the residue and the mixture
was extracted with EtOAc. The organics were dried over
Na.sub.2SO.sub.4 and the solvent was evaporated under vacuum to
obtain 0.87 g of crude product
2-tert-butyl-S-nit-1H-indole-7-carbonitrile which was used directly
in the next step without purification.
##STR00594##
5-Amino-2-tert-butyl-1H-indol-7-carbonitrile
[0411] To a solution of crude product
2-tert-butyl-5-nitro-1H-indole-7-carbonitrile (0.87 g, 3.6 mmol) in
MeOH (10 mL) was added NiCl.sub.2.6H.sub.2O (1.8 g, 7.2 mmol) at
-5.degree. C. The reaction mixture was stirred for 30 min, then
NaBH.sub.4 (0.48 g, 14.32 mmol) was added to the reaction mixture
at 0.degree. C. After 5 min, the reaction mixture was quenched with
water, filtered and extracted with EtOAc. The combined organic
layers were dried over Na.sub.2SO.sub.4 and concentrated under
vacuum to obtain the crude product, which was purified by column
chromatography (5-20% EtOAc in petroleum ether) to obtain
5-amino-2-tert-butyl-1H-indol-7-carbonitrile (470 mg, 32% over two
steps). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.25 (s, 1H),
7.06 (d, J=2.4 Hz, 1H), 6.84 (d, J=2.4 Hz, 1H), 6.14 (d, J=2.4 Hz,
1H), 3.57 (br s, 2H), 1.38 (s, 9H). MS (ESI) m/z: 214
(M+H.sup.+).
Example 40
Methyl 5-amino-2-tert-butyl-1H-indole-7-carboxylate
##STR00595##
[0412] 2-tert-Butyl-5-nitro-1H-indole-7-carboxylic acid
[0413] 2-tert-Butyl-5-nitro-1H-indole-7-carbonitrile (4.6 g, 19
mmol) was added to a solution of KOH in EtOH (10%, 100 mL) and the
mixture was heated at reflux overnight. The solution was evaporated
to remove alcohol, a small amount of water was added, and then the
mixture was acidified with dilute hydrochloric acid. Upon standing
in the refrigerator, an orange-yellow solid precipitated, which was
purified by chromatography on silica gel (15% EtOAc in petroleum
ether) to afford 2-tert-butyl-5-nitro-1H-indole-7-carboxylic acid
(4.0 g, 77%). .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 10.79 (brs,
1H), 8.66 (s, 1H), 8.45 (s, 1H), 6.57 (s, I H), 1.39 (s, 9H).
##STR00596##
Methyl 2-tert-butyl-5-nitro-1H-indole-7-carboxylate
[0414] SOCl.sub.2 (3.6 g, 30 mol) was added dropwise to a solution
of 2-tert-butyl-5-nitro-1H-indole-7-carboxylic acid (4.0 g, 15 mol)
and methanol (30 mL) at 0.degree. C. The mixture was stirred at
80.degree. C. for 12 h. The solvent was evaporated under vacuum and
the residue was purified by column chromatography on silica gel (5%
EtOAc in petroleum ether) to afford methyl
2-tert-butyl-5-nitro-1H-indole-7-carboxylate (2.95 g, 70%). .sup.1H
NMR (CDCl.sub.3, 300 MHz) .delta. 9.99 (brs, 1H), 8.70 (d, J=2.1
Hz, 1H), 8.65 (d, J=2.1 Hz, 1H), 6.50 (d, J=2.4 Hz, 1H), 4.04 (s,
3H), 1.44 (s, 9H).
##STR00597##
Methyl 5-amino-2-tert-butyl-1H-indole-7-carboxylate
[0415] A solution of 2-tert-butyl-5-nitro-1H-indole-7-carboxylate
(2.0 g, 7.2 mmol) and Raney Nickel (200 mg) in CH.sub.3OH (50 mL)
was stirred for 5 h at the room temperature under H.sub.2
atmosphere. The catalyst was filtered off through a celite pad and
the filtrate was evaporated under vacuum to give methyl
5-amino-2-tert-butyl-1H-indole-7-carboxylate (1.2 g, 68%) .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 9.34 (brs, 1H), 7.24 (d, J=1.6
Hz, 1H), 7.10 (s, 1H), 6.12 (d, J=1.6 Hz, 1H), 3.88 (s, 3H), 1.45
(s, 9H).
Example 41
(5-Amino-2-tert-butyl-1H-indol-7-yl)methanol
##STR00598##
[0416] (2-tert-Butyl-5-nitro-1H-indol-7-yl) methanol
[0417] To a solution of methyl
2-tert-butyl-5-nitro-1H-indole-7-carboxylate (6.15 g, 22.3 mmol)
and dichloromethane (30 ml) was added DIBAL-H (1.0 M, 20 mL, 20
mmol) at 78.degree. C.
[0418] The mixture was stirred for 1 h before water (10 mL) was
added slowly. The resulting mixture was extracted with EtOAc (120
mL.times.3). The combined organic extracts were dried over
anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum to give
(2-tert-butyl-5-nitro-1H-indol-7-yl)methanol (4.0 g, 73%), which
was used in the next step directly.
##STR00599##
(5-Amino-2-tert-butyl-1H-indol-7-yl)methanol
[0419] A mixture of (2-tert-butyl-5-nitro-1H-indol-7-yl)methanol
(4.0 g, 16 mmol) and Raney Nickel (400 mg) in CH.sub.3OH (100 mL)
was stirred for 5 g at room temperature under H.sub.2. The catalyst
was filtered off through a celite pad and the filtrate was
evaporated under vacuum to give
(5-amino-2-tert-butyl-1H-indol-7-yl)methanol (3.4 g, 80%). .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.53 (br s, 1H), 6.80 (d, J=2.0
Hz, 1H), 6.38 (d, J=1.6 Hz, 1H), 4.89 (s, 2H), 1.37 (s, 9H).
Example 42
2-(1-Methylcyclopropyl)-1H-indol-5-amine
##STR00600##
[0420] Trimethyl-(1-methyl-cyclopropylethynyl)-silane
[0421] To a solution of cyclopropylethynyl-trimethyl-silane (3.0 g,
22 mmol) in ether (20 mL) was added dropwise n-BuLi (8.6 mL, 21.7
mol, 2.5 M solution in hexane) at 0.degree. C. The reaction mixture
was stirred at ambient temperature for 24 h before dimethyl sulfate
(6.85 g, 54.3 mmol) was added dropwise at -10.degree. C. The
resulting solution was stirred at 10.degree. C. and then at
20.degree. C. for 30 min each. The reaction was quenched by adding
a mixture of sat. aq. NH.sub.4Cl and 25% aq. ammonia (1:3, 100 mL).
The mixture was then stirred at ambient temperature for 1 h. The
aqueous phase was extracted with diethyl ether (3.times.50 mL) and
the combined organic layers were washed successively with 5%
aqueous hydrochloric acid (100 mL), 5% aq. NaHCO.sub.3 solution
(100 mL), and water (100 mL). The organics were dried over
anhydrous NaSO.sub.4 and concentrated at ambient pressure. After
fractional distillation under reduced pressure,
trimethyl-(1-methyl-cyclopropylethynyl)-silane (1.7 g, 52%) was
obtained as a colorless liquid. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 1.25 (s, 3H), 0.92-0.86 (m, 2H), 0.58-0.56 (m, 2H), 0.15
(s, 9H).
##STR00601##
1-Ethynyl-1-methyl-cyclopropane
[0422] To a solution of
trimethyl-(l-methyl-cyclopropylethynyl)-silane (20 g, 0.13 mol) in
THF (250 mL) was added TBAF (69 g, 0.26 mol). The mixture was
stirred overnight at 20.degree. C. The mixture was poured into
water and the organic layer was separated. The aqueous phase was
extracted with THF (50 mL). The combined organic layers were dried
over anhydrous Na.sub.2SO.sub.4 and distilled under atmospheric
pressure to obtain 1-ethynyl-1-methyl-cyclopropane (7.0 g,
contained 1/2 THF, 34%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
1.82 (s, 1H), 1.26 (s, 3H), 0.90-0.88 (m, 2H), 0.57-0.55 (m,
2H).
##STR00602##
2-Bromo-4-nitroaniline
[0423] To a solution of 4-nitro-phenylamine (50 g, 0.36 mol) in
AcOH (500 mL) was added Br.sub.2 (60 g, 0.38 mol) dropwise at
5.degree. C. The mixture was stirred for 30 min at that
temperature. The insoluble solid was collected by filtration and
basified with saturated aqueous NaHCO.sub.3 to pH 7. The aqueous
phase was extracted with EtOAc (300 mL.times.3). The combined
organic layers were dried and evaporated under reduced pressure to
obtain compound 2-bromo-4-nitroaniline (56 g, 72%), which was
directly used in the next step.
##STR00603##
2-((1-Methylcyclopropyl)ethynyl)-4-nitroaniline
[0424] To a deoxygenated solution of 2-bromo-4-nitroaniline (430
mg, 2.0 mmol) and 1-ethynyl-1-methyl-cyclopropane (630 mg, 8.0
mmol) in triethylamine (20 mL) was added CuI (76 mg, 0.40 mmol) and
Pd(PPh.sub.3).sub.2Cl.sub.2 (140 mg, 0.20 mmol) under N.sub.2. The
mixture was heated at 70.degree. C. and stirred for 24 h. The solid
was filtered off and washed with EtOAc (50 mL.times.3). The
filtrate was evaporated under reduced pressure and the residue was
purified by column chromatography on silica gel (petroleum
ether/ethyl acetate=10/1) to give
2-((1-methylcyclopropyl)ethynyl)-4-nitroaniline (340 mg, 79%).
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.15-8.14 (m, 1H),
7.98-7.95 (m, 1H), 6.63 (d, J=6.9 Hz, 1H), 4.80 (brs, 2H), 1.38 (s,
3H), 1.04-1.0 (m, 2H), 0.76-0.73 (m, 2H).
##STR00604##
N-[2-(1-Methyl-cyclopropylethynyl)-4-nitro-phenyl]-butyramide
[0425] To a solution of
2-((1-methylcyclopropyl)ethynyl)-4-nitroaniline (220 mg, 1.0 mmol)
and pyridine (160 mg, 2.0 mol) in CH.sub.2Cl.sub.2 (20 mL) was
added butyryl chloride (140 mg, 1.3 mmol) at 0.degree. C. The
mixture was warmed to room temperature and stirred for 3 h. The
mixture was poured into ice-water. The organic layer was separated
and the aqueous phase was extracted with CH.sub.2Cl.sub.2 (30
mL.times.3). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under reduced pressure to obtain
N-[2-(1-methyl-cyclopropylethynyl)-4-nitro-phenyl]-butyramide (230
mg, 82%), which was directly used in the next step.
##STR00605##
2-(1-Methylcyclopropyl)-5-nitro-1H-indole
[0426] A mixture of
N-[2-(1-methyl-cyclopropylethynyl)-4-nitro-phenyl]-butyramide (1.3
g, 4.6 mmol) and TBAF (2.4 g, 9.2 mmol) in THF (20 mL) was heated
at reflux for 24 h. The mixture was cooled to room temperature and
poured into ice water. The mixture was extracted with
CH.sub.2Cl.sub.2 (30 mL.times.3). The combined organic layers were
dried over anhydrous Na.sub.2SO.sub.4 and evaporated under reduced
pressure. The residue was purified by column chromatography on
silica gel (petroleum ether/ethyl acetate=10/1) to afford
2-(1-methylcyclopropyl)-5-nitro-1H-indole (0.70 g, 71%). .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 8.56 (brs, 1H), 8.44 (d, J=2.0
Hz, 1H), 8.01 (dd, J=2.4, 8.8 Hz, 1H), 7.30 (d, J=8.8 Hz, 1H), 6.34
(d, J=1.6 Hz, 1H), 1.52 (s, 3H), 1.03-0.97 (m, 2H), 0.89-0.83 (m,
2H).
##STR00606##
2-(1-Methyl-cyclopropyl)-1H-indol-5-ylamine
[0427] To a solution of 2-(1-methylcyclopropyl)-5-nitro-1H-indole
(0.70 g, 3.2 mmol) in EtOH (20 mL) was added Raney Nickel (100 mg)
under nitrogen atmosphere. The mixture was stirred under hydrogen
atmosphere (1 atm) at room temperature overnight. The catalyst was
filtered off through a celite pad and the filtrate was evaporated
under vacuum. The residue was purified by column chromatography on
silica gel (petroleum ether/ethyl acetate=5/1) to afford
2-(1-methyl-cyclopropyl)-1H-indol-5-ylamine (170 mg, 28%). .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.65 (brs, 1H), 7.08 (d, J=8.4
Hz, 1H), 6.82 (s, 1H), 6.57 (d, J=8.4 Hz, 1H), 6.14 (s, 1H), 3.45
(brs, 2H), 1.47 (s, 3H), 0.82-0.78 (m, 2H), 0.68-0.63 (m, 2H).
Example 43
Methyl 2-(5-amino-1H-indol-2-yl)-2-methylpropanoate
##STR00607## ##STR00608##
[0428] Methyl 2,2-dimethyl-3-oxobutanoate
[0429] To a suspension of NaH (42 g, 1.1 mol, 60%) in THF (400 mL)
was added dropwise a solution of methyl 3-oxobutanoate (116 g, 1.00
mol) in THF (100 mL) at 0.degree. C. The mixture was stirred for
0.5 h at that temperature before MeI (146 g, 1.1 mol) was added
dropwise at 0.degree. C. The resultant mixture was warmed to room
temperature and stirred for 1 h. NaH (42 g, 1.05 mol, 60%) was
added in portions at 0.degree. C. and the resulting mixture was
continued to stir for 0.5 h at this temperature. MeI (146 g, 1.05
mol) was added dropwise at 0.degree. C. The reaction mixture was
warmed to room temperature and stirred overnight. The mixture was
poured into ice water and the organic layer was separated. The
aqueous phase was extracted with EtOAc (500 mL.times.3). The
combined organic layers were dried and evaporated under reduced
pressure to give methyl 2,2-dimethyl-3-oxobutanoate (85 g), which
was used directly in the next step.
##STR00609##
Methyl 3-chloro-2,2-dimethylbut-3-enoate
[0430] To a suspention of PCl.sub.5 (270 g, 1.3 mol) in
CH.sub.2Cl.sub.2 (1000 mL) was added dropwise methyl
2,2-dimethyl-3-oxobutanoate (85 g) at 0.degree. C., following by
addition of approximately 30 drops of dry DMF. The mixture was
heated at reflux overnight. The reaction mixture was cooled to
ambient temperature and slowly poured into ice water. The organic
layer was separated and the aqueous phase was extracted with
CH.sub.2Cl.sub.2 (500 mL.times.3). The combined organic layers were
washed with saturated aqueous NaHCO.sub.3 and dried over anhydrous
Na.sub.2SO.sub.4. The solvent was evaporated and the residue was
distilled under reduced pressure to give methyl
3-chloro-2,2-dimethylbut-3-enoate (37 g, 23%). .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 5.33 (s, 1H), 3.73 (s, 3H), 1.44 (s,
6H).
##STR00610##
3-Chloro-2,2-dimethylbut-3-enoic acid
[0431] A mixture of methyl 3-chloro-2,2-dimethylbut-3-enoate (33 g,
0.2 mol) and NaOH (9.6 g, 0.24 mol) in water (200 mL) was heated at
reflux for 5 h. The mixture was cooled to ambient temperature and
extracted with ether. The organic layer was discarded. The aqueous
layer was acidified with cold 20% HCl solution and extracted ether
(200 mL.times.3). The combined organic layers were dried and
evaporated under reduced pressure to give
3-chloro-2,2-dimethyl-but-3-enoic acid (21 g, 70%), which was used
directly in the next step. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.90 (brs, 1H), 5.37 (dd, J=2.4, 6.8 Hz, 2H), 1.47 (s,
6H).
##STR00611##
2,2-Dimethyl-but-3-ynoic acid
[0432] Liquid NH.sub.3 was condensed in a 3-neck, 250 mL round
bottom flask at -78.degree. C. Na (3.98 g, 0.173 mol) was added to
the flask in portions. The mixture was stirred for 2 h at
-78.degree. C. before anhydrous DMSO (20 mL) was added dropwise at
-78.degree. C. The mixture was stirred at room temperature until no
more NH.sub.3 was given off. A solution of
3-chloro-2,2-dimethyl-but-3-enoic acid (6.5 g, 43 mmol) in DMSO (10
mL) was added dropwise at -40.degree. C. The mixture was warmed and
stirred at 50.degree. C. for 5 h, then stirred at room temperature
overnight. The cloudy, olive green solution was poured into cold
20% HCl solution and then extracted three times with ether. The
ether extracts were dried over anhydrous Na.sub.2SO.sub.4 and
concentrated to give crude 2,2-dimethyl-but-3-ynoic acid (2 g),
which was used directly in the next step. .sup.1H NMR (400 M z,
CDCl.sub.3) .delta. 2.30 (s, 1H), 1.52 (s, 6H).
##STR00612##
Methyl 2,2-dimethylbut-3-ynoate
[0433] To a solution of diazomethane (.about.10 g) in ether (400
mL) was added dropwise 2,2-dimethyl-but-3-ynoic acid (10.5 g, 93.7
mmol) at 0.degree. C. The mixture was warmed to room temperature
and stirred overnight. The mixture was distilled under atmospheric
pressure to give crude methyl 2,2-dimethylbut-3-ynoate (14 g),
which was used directly in the next step. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 3.76 (s, 3H), 2.28 (s, 1H), 1.50 (s, 6H).
##STR00613##
Methyl 4-(2-amino-S-nitrophenyl)-2,2-dimethylbut-3-ynoate
[0434] To a deoxygenated solution of compound
2-bromo-4-nitroaniline (9.43 g, 43.7 mmol), methyl
2,2-dimethylbut-3-ynoate (5.00 g, 39.7 mmol), CuI (754 mg, 3.97
mmol) and triethylamine (8.03 g, 79.4 mmol) in toluene/H.sub.2O
(100/30 mL) was added Pd(PPh.sub.3).sub.4 (6.17 g, 3.97 mmol) under
N.sub.2. The mixture was heated at 70.degree. C. and stirred for 24
h. After cooling, the solid was filtered off and washed with EtOAc
(50 mL.times.3). The organic layer was separated and the aqueous
phase was washed with EtOAc (50 mL.times.3). The combined organic
layers were dried and evaporated under reduced pressure to give a
residue, which was purified by column chromatography on silica gel
(petroleum ether/ethyl acetate=10/1) to obtain methyl
4-(2-amino-5-nitrophenyl)-2,2-dimethylbut-3-ynoate (900 mg, 9%).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.17 (d, J=2.8 Hz, 1H),
8.01 (dd, J=2.8, 9.2 Hz, 1H), 6.65 (d, J=9.2 Hz, 1H), 5.10 (brs,
2H), 3.80 (s, 3H), 1.60 (s, 6H).
##STR00614##
Methyl 4-(2-butyramido-5-nitrophenyl)-2,2-dimethylbut-3-ynoate
[0435] To a solution of methyl
4-(2-amino-5-nitrophenyl)-2,2-dimethylbut-3-ynoate (260 mg, 1.0
mmol) and pyridine (160 mg, 2.0 mol) in CH.sub.2Cl.sub.2 (20 mL)
was added butyryl chloride (140 mg, 1.3 mmol) at 0.degree. C. The
reaction mixture was warmed to room temperature and stirred for 3 h
before the mixture was poured into ice-water. The organic layer was
separated and the aqueous phase was extracted with CH.sub.2Cl.sub.2
(30 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4 and evaporated under reduced pressure to
obtain methyl
4-(2-butyramido-5-nitrophenyl)-2,2-dimethylbut-3-ynoate (150 mg,
45%), which was used directly in the next step. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 8.79 (brs, 1H), 8.71 (d, J=9.2 Hz, 1H),
8.24 (d, J=2.8 Hz, 1H), 8.17 (dd, J=2.8, 9.2 Hz, 1H), 3.82 (s, 3H),
2.55 (t, J=7.2 Hz, 2H), 1.85-1.75 (m. 2H), 1.63 (s, 6H), 1.06 (t,
J=6.8 Hz, 3H).
##STR00615##
Methyl 2-methyl-2-(5-nitro-1H-indol-2-yl)propanoate
[0436] To a deoxygenated solution of methyl
4-(2-butyramido-5-nitrophenyl)-2,2-dimethylbut-3-ynoate (1.8 g, 5.4
mmol) in acetonitrile (30 mL) was added
Pd(CH.sub.3CN).sub.2Cl.sub.2 (0.42 g, 1.6=mmol) under N.sub.2. The
mixture was heated at reflux for 24 h. After cooling the mixture to
ambient temperature, the solid was filtered off and washed with
EtOAc (50 mL.times.3). The filtrate was evaporated under reduced
pressure to give a residue, which was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate=30/1)
to give methyl 2-methyl-2-(5-nitro-1H-indol-2-yl)propanoate (320
mg, 23%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.05 (brs, 1H),
8.52 (d, J=2.0 Hz, 1H), 8.09 (dd, J=2.0, 8.8 Hz, 1H), 7.37 (d,
J=8.8 Hz, 1H), 6.54 (d, J=1.6 Hz, 1H), 3.78 (d, J=9.6 Hz, 3H), 1.70
(s, 6H).
##STR00616##
Methyl 2-(5-amino-1H-indol-2-yl)-2-methylpropanoate
[0437] A suspension of methyl
2-methyl-2-(5-nitro-1H-indol-2-yl)propanoate (60 mg, 0.23 mmol) and
Raney Nickel (10 mg) in MeOH (5 mL) was hydrogenated under hydrogen
(1 atm) at room temperature overnight. The catalyst was filtered
off through a celite pad and the filtrate was evaporated under
vacuum to give a residue, which was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate=5/1) to
give methyl 2-(5-amino-1H-indol-2-yl)-2-methylpropanoate (20 mg,
38%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.37 (br s, 1H),
7.13 (d, J=8.4 Hz, 1H), 6.87 (d, J=2.0 Hz, 1H), 6.63 (dd, J=2.0,
8.4 Hz, 1H), 6.20 (d, J=1.2 Hz, 1H), 3.72 (d, J=7.6 Hz, 3H), 3.43
(br s, 1H), 1.65 (s, 6H); MS (ESI) m/e (M+H.sup.+) 233.2.
Example 44
2-Isopropyl-1H-indol-5-amine
##STR00617##
##STR00618##
[0438] 2-Isopropyl-5-nitro-1H-indole
[0439] A mixture of methyl
4-(2-butyramido-5-nitrophenyl)-2,2-dimethylbut-3-ynoate (0.50 g,
1.5 mmol) and TBAF (790 mg, 3.0 mmol) in DMF (20 mL) was heated at
70.degree. C. for 24 h. The reaction mixture was cooled to room
temperature and poured into ice water. The mixture was extracted
with ether (30 mL.times.3). The combined organic layers were dried
over anhydrous Na.sub.2SO.sub.4 and evaporated under reduced
pressure to give a residue, which was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate=20/1)
to give 2-isopropyl-5-nitro-1H-indole (100 mg, 33%). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.68 (s, 1H), 8.25 (br s, 1H), 8.21
(dd, J=2.4, 10.0 Hz, 1H), 7.32 (d, J=8.8 Hz, 1H), 6.41 (s, 1H),
3.07-3.14 (m, 1H), 1.39 (d, J=6.8 Hz, 6H).
##STR00619##
2-Isopropyl-1H-indol-5-amine
[0440] A suspension of 2-isopropyl-5-nitro-1H-indole (100 mg, 0.49
mmol) and Raney Nickel (10 mg) in MeOH (10 mL) was hydrogenated
under hydrogen (1 atm) at the room temperature overnight. The
catalyst was filtered off through a celite pad and the filtrate was
evaporated under vacuum to give a residue, which was purified by
column (petroleum ether/ethyl acetate=5/1) to give
2-isopropyl-1H-indol-5-amine (35 mg, 41%). .sup.1H NMR (400 MHz,
CDCl.sub.3) b 7.69 (br s, 1H), 7.10 (d, J=8.4 Hz, 1H), 6.86 (d,
J=2.4 Hz, 1H), 6.58 (dd, J=2.4, 8.8 Hz, 1H), 6.07 (t, J=1.2 Hz,
1H), 3.55 (br s, 2H), 3.06-2.99 (m, 1H), 1.33 (d, J=7.2 Hz, 6H); MS
(ESI) m/e (M+H.sup.+) 175.4.
Example 45
1-(Benzo[d][1,3]dioxo-5-yl)-N-(2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol--
5-yl)cyclopropanecarboxamide
##STR00620##
[0441] Triphenyl(2-aminobenzyl)phosphonium bromide
[0442] 2-Aminobenzyl alcohol (60.0 g, 0.487 mol) was dissolved in
acetonitrile (2.5 L) and brought to reflux. Triphenylphosphine
hydrobromide (167 g, 0.487 mol) was added and the mixture was
heated at reflux for 3 h. The reaction mixture was concentrated to
approximately 500 mL and left at room temperature for 1 h. The
precipitate was filtered and washed with cold acetonitrile followed
by hexane. The solid was dried overnight at 40.degree. C. under
vacuum to give triphenyl(2-aminobenzyl)phosphonium bromide (193 g,
88%).
##STR00621##
Triphenyl((ethyl(2-carbamoyl)acetate)-2-benzyl)phosphonium
bromide
[0443] To a suspension of triphenyl(2-aminobenzyl)phosphonium
bromide (190 g, 0.43 mol) in anhydrous dichloromethane (1 L) was
added ethyl malonyl chloride (55 ml, 0.43 mol). The reaction was
stirred for 3 h at room temperature. The mixture was evaporated to
dryness before ethanol (400 mL) was added. The mixture was heated
at reflux until a clear solution was obtained. The solution was
left at room temperature for 3 h. The precipitate was filtered,
washed with cold ethanol followed by hexane and dried. A second
crop was obtained from the mother liquor in the same way. In order
to remove residual ethanol both crops were combined and dissolved
in dichloromethane (approximately 700 mL) under heating and
evaporated. The solid was dried overnight at 50.degree. C. under
vacuum to give
triphenyl((ethyl(2-carbamoyl)acetate)-2-benzyl)-phosphonium bromide
(139 g, 58%).
##STR00622##
Ethyl 2-(1H-indol-2-yl)acetate
[0444] Triphenyl((ethyl(2-carbamoyl)acetate)-2-benzyl)phosphonium
bromide (32.2 g, 57.3 mmol) was added to anhydrous toluene (150 mL)
and the mixture was heated at reflux. Fresh potassium tert-butoxide
(7.08 g, 63.1 mmol) was added in portions over 15 minutes. Reflux
was continued for another 30 minutes. The mixture was filtered hot
through a plug of celite and evaporated under reduced pressure. The
residue was purified by column chromatography on silica gel (0-30%
ethyl acetate in hexane over 45 min) to give ethyl
2-(1H-indol-2-yl)acetate (9.12 g, 78%).
##STR00623##
tert-Butyl 2-((ethoxycarbonyl)methyl)-1H-indole-1-carboxylate
[0445] To a solution of ethyl 2-(1H-indol-2-yl)acetate (14.7 g,
72.2 mmol) in dichloromethane (150 mL) was added
4-dimethylaminopyridine (8.83 g, 72.2 mmol) and di-tert-butyl
carbonate (23.7 g, 108 mmol) in portions. After stirring for 2 h at
room temperature, the mixture was diluted with dichloromethane,
washed with water, dried over magnesium sulfate and purified by
silica gel chromatography (0 to 20% EtOAc in hexane) to give
tert-butyl 2-((ethoxycarbonyl)methyl)-1H-indole-carboxylate (20.0
g, 91%).
##STR00624##
tert-Butyl
2-(2-(ethoxycarbonyl)propan-2-yl)-1H-indole-1-carboxylate
[0446] tert-Butyl 2-((ethoxycarbonyl)methyl)-1H-indole-carboxylate
(16.7 g, 54.9 mmol) was added to anhydrous THF (100 mL) and cooled
to -78.degree. C. A 0.5M solution of potassium hexamethyldisilazane
(165 mL, 82 mmol) was added slowly such that the internal
temperature stayed below -60.degree. C. Stirring was continued for
30 minutes at -78.degree. C. To this mixture, methyl iodide (5.64
mL, 91 mmol) was added. The mixture was stirred for 30 min at room
temperature and then cooled to -78.degree. C. A 0.5M solution of
potassium hexamethyldisilazane (210 mL, 104 mmol) was added slowly
and the mixture was stirred for another 30 minutes at -78.degree.
C. More methyl iodide (8.6 mL, 137 mmol) was added and the mixture
was stirred for 1.5 h at room temperature. The reaction was
quenched with sat. aq. ammonium chloride and partitioned between
water and dichloromethane. The aqueous phase was extracted with
dichloromethane and the combined organic phases were dried over
magnesium sulfate and evaporated under reduced pressure. The
residue was purified by column chromatography on silica gel (0 to
20% ethylacetate in hexane) to give tert-butyl
2-(2-(ethoxycarbonyl)propan-2-yl)-1H-indole-1-carboxylate (17.1 g,
94%).
##STR00625##
Ethyl 2-(1H-indol-2-yl)-2-methylpropanoate
[0447] tert-Butyl
2-(2-(ethoxycarbonyl)propan-2-yl)-1H-indole-1-carboxylate (22.9 g,
69.1 mmol) was dissolved in dichloromethane (200 mL) before TFA (70
mL) was added. The mixture was stirred for 5 h at room temperature.
The mixture was evaporated to dryness, taken up in dichloromethane
and washed with saturated sodium bicarbonate solution, water, and
brine. The product was purified by column chromatography on silica
gel (0-20% EtOAc in hexane) to give ethyl
2-(1H-indol-2-yl)-2-methylpropanoate (12.5 g, 78%).
##STR00626##
Ethyl 2-methyl-2-(5-nitro-1H-indol-2-yl)propanoate
[0448] Ethyl 2-(1H-indol-2-yl)-2-methylpropanoate (1.0 g, 4.3 mmol)
was dissolved in concentrated sulfuric acid (6 mL) and cooled to
-10.degree. C. (salt/ice-mixture). A solution of sodium nitrate
(370 mg, 4.33 mmol) in concentrated sulfuric acid (3 mL) was added
dropwise over 30 min. Stirring was continued for another 30 min at
-10.degree. C. The mixture was poured into ice and the product was
extracted with dichloromethane. The combined organic phases were
washed with a small amount of sat. aq. sodium bicarbonate. The
product was purified by column chromatography on silica gel (5-30%
EtOAc in hexane) to give ethyl
2-methyl-2-(5-nitro-1H-indol-2-yl)propanoate (0.68 g, 57%).
##STR00627##
2-Methyl-2-(5-nitro-1H-indol-2-yl)propan-1-ol
[0449] To a cooled solution of LiAlH.sub.4 (1.0 M in THF, 1.1 mL,
1.1 mmol) in THF (5 mL) at 0.degree. C. was added a solution of
ethyl 2-methyl-2-(5-nitro-1H-indol-2-yl)propanoate (0.20 g, 0.72
mmol) in THF (3.4 mL) dropwise. After addition, the mixture was
allowed to warm up to room temperature and was stirred for 3 h. The
mixture was cooled to 0.degree. C. before water (2 mL) was slowly
added followed by careful addition of 15% NaOH (2 mL) and water (4
mL). The mixture was stirred at room temperature for 0.5 h and was
filtered through a short plug of celite using ethyl acetate. The
organic layer was separated from the aqueous layer, dried over
Na.sub.2SO.sub.4, filtered and evaporated under reduced pressure.
The residue was purified by column chromatography on silica gel
(ethyl acetate/hexane=1/1) to give
2-methyl-2-(5-nitro-1H-indol-2-yl)propan-1-ol (0.098 g, 58%).
##STR00628##
2-(5-Amino-1H-indol-2-yl)-2-methylpropan-1-ol
[0450] To a solution of
2-methyl-2-(5-nitro-1H-indol-2-yl)propan-1-ol (0.094 g, 0.40 mmol)
in ethanol (4 mL) was added tin chloride dihydrate (0.451 g, 2.0
mmol). The mixture was heated in the microwave at 120.degree. C.
for 1 h. The mixture was diluted with ethyl acetate and water
before being quenched with saturated aqueous NaHCO.sub.3. The
reaction mixture was filtered through a plug of celite using ethyl
acetate. The organic layer was separated from the aqueous layer,
dried over Na.sub.2SO.sub.4, filtered and evaporated under reduced
pressure to give 2-(5-amino-1H-indol-2-yl)-2-methylpropan-1-ol
(0.080 g, 98%).
Example 46
2-(Pyridin-2-yl)-1H-indol-5-amine
##STR00629##
[0451] 4-Nitro-2-(pyridin-2-ylethynyl)aniline
[0452] To the solution of 2-iodo-4-nitroaniline (3.0 g, 11 mmol) in
DMF (60 mL) and Et.sub.3N (60 mL) was added 2-ethynylpyridine (3.0
g, 45 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (600 mg) and CuI (200 mg)
under N.sub.2. The reaction mixture was stirred at 60.degree. C.
for 12 h. The mixture was diluted with water and extracted with
dichloromethane (3.times.100 mL). The combined organic layers were
washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuum. The residue was purified by chromatography
on silica gel (5-10% ethyl acetate/petroleum ether) to afford
4-nitro-2-(pyridin-2-ylethynyl)aniline (1.5 g, 60%). .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 8.60 (s, 1H), 8.13 (d, J=2.1 Hz, 1H),
7.98 (d, J=1.8, 6.9 Hz, 1H), 7.87-7.80 (m, 2H), 7.42-7.39 (m, 1H),
7.05 (brs, 2H), 6.80 (d, J=6.9 Hz, 1H).
##STR00630##
5-Nitro-2-(pyridin-2-yl)-1H-indole
[0453] To the solution of 4-nitro-2-(pyridin-2-ylethynyl)aniline
(1.5 g, 6.3 mmol) in DMF (50 mL) was added t-BuOK (1.5 g, 13 mmol).
The reaction mixture was stirred at 90.degree. C. for 2 h. The
mixture was diluted with water and extracted with dichloromethane
(3.times.50 mL). The combined organic layers were washed with
brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated in
vacuum. The residue was purified by chromatography on silica gel
(5-10% ethyl acetate/petroleum ether) to afford
5-nitro-2-(pyridin-2-yl)-1H-indole (1.0 g, 67% yield). .sup.1H NMR
(300 MHz, d-DMSO) .delta. 12.40 (s, 1H), 8.66 (d, J=2.1 Hz, 1H),
8.58 (d, J=1.8 Hz, 1H), 8.07-7.91 (m, 3H), 7.59 (d, J=6.6 Hz, 1H),
7.42-7.37 (m, 2H).
##STR00631##
2-(Pyridin-2-yl)-1H-indol-5-amine
[0454] To a solution of 5-nitro-2-(pyridin-2-yl)-1H-indole (700 mg,
2.9 mmol) in EtOH (20 mL) was added SnCl.sub.2 (2.6 g, 12 mmol).
The mixture was heated at reflux for 10 h. Water was added and the
mixture was extracted with EtOAc (50 mL.times.3). The combined
organic layers were washed with brine, dried over anhydrous
Na.sub.2SO.sub.4 and concentrated in vacuum. The residue was
purified by chromatography on silica gel (5-10% ethyl
acetate/petroleum ether) to afford
2-(pyridin-2-yl)-1H-indol-5-amine (120 ag, 20%). .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 9.33 (brs, 1H), 8.55 (dd, J=1.2, 3.6 Hz,
1H), 7.76-7.67 (m, 2H), 7.23 (d, J=6.4 Hz, 1H), 7.16-7.12 (m, 1H),
6.94 (d, J=2.0 Hz, 1H), 6.84 (d, J=2.4 Hz, 1H), 6.71-6.69 (dd,
J=2.0, 8.4 Hz, 1H).
Example 47
2-(Pyridin-2-yl)-1H-indol-5-amine
##STR00632## ##STR00633##
[0455]
[2-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-(2-iodo-4-nitro-phenyl)--
amine
[0456] To a solution of 2-iodo-4-nitroaniline (2.0 g, 7.6 mmol) and
2-(tert-butyldimethylsilyloxy)-acetaldehyde (3.5 g, 75% purity, 15
mmol) in methanol (30 mL) was added TFA (1.5 mL) at 0.degree. C.
The reaction mixture was stirred at this temperature for 30 min
before NaCNBH.sub.3 (900 mg, 15 mmol) was added in portions. The
mixture was stirred for 2 h and was then quenched with water. The
resulting mixture was extracted with EtOAc (30 mL.times.3), the
combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under vacuum, and the residue was
purified by chromatography on silica gel (5% ethyl
acetate/petroleum) to afford
(2-(tert-butyl-dimethyl-silanyloxy)-ethyl]-(2-iodo-4-nitro-phen-
yl)-amine (800 mg, 25%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
8.57 (d, J=2.7 Hz, 1H), 8.12 (dd, J=2.4, 9.0 Hz, 1H), 6.49 (d,
J=9.3 Hz, 1H), 5.46 (br s, 1H), 3.89 (t, J=5.4 Hz, 2H), 3.35 (q,
J=5.4 Hz, 2H), 0.93 (s, 9H), 0.10 (s, 6H).
##STR00634##
5-{2-[2-(tert-Butyl-dimethyl-silanyloxy)-ethylamino]-5-nitro-phenyl}-3,3--
dimethyl-pent-4-ynoic acid ethyl ester
[0457] To a solution of
[2-(tert-butyl-dimethyl-silanyloxy)-ethyl]-(2-iodo-4-nitro-phenyl)-amine
(800 mg, 1.9 mmol) in Et.sub.3N (20 mL) was added
Pd(PPh.sub.3).sub.2Cl.sub.2 (300 mg, 0.040 mmol), CuI (76 mg, 0.040
mmol) and 3,3-dimethyl-but-1-yne (880 mg, 5.7 mmol) successively
under N.sub.2 protection. The reaction mixture was heated at
80.degree. C. for 6 h and allowed to cool down to room temperature.
The resulting mixture was extracted with EtOAc (30 mL.times.3). The
combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4 and evaporated under vacuum to give
5-{2-[2-(tert-butyl-dimethyl-silanyloxy)-ethylamino]-5-nitro-phenyl}-3,3--
dimethyl-pent-4-ynoic acid ethyl ester (700 mg, 82%), which was
used in the next step without further purification. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.09 (s, 1H), 8.00 (d, J=9.2 Hz, 1H),
6.54 (d, J=9.2 Hz, 1H), 6.45 (brs, 1H), 4.17-4.10 (m, 4H), 3.82 (t,
J=5.6 Hz, 2H), 3.43 (q, J=5.6 Hz, 2H), 2.49 (s, 2H), 1.38 (s, 6H),
1.28 (t, J=7.2 Hz, 3H), 0.84 (s, 9H), 0.00 (s, 6H).
##STR00635##
3-[1-(2-Hydroxy-ethyl)-5-nitro-1H-indol-2-yl]-3-methyl-butyric acid
ethyl ester
[0458] A solution of
5-{2-[2-(tert-butyl-dimethyl-silanyloxy)-ethylamino]-5-nitro-phenyl}-3,3--
dimethyl-pent-4-ynoic acid ethyl ester (600 mg, 1.34 mmol) and
PdCl.sub.2 (650 mg) in CH.sub.3CN (30 mL) was heated at reflux
overnight. The resulting mixture was extracted with EtOAc (30
mL.times.3). The combined organic extracts were dried over
anhydrous Na.sub.2SO.sub.4 and evaporated under vacuum. The residue
was dissolved in THF (20 mL) and TBAF (780 mg, 3.0 mmol) was added.
The mixture was stirred at room temperature for 1 h, the solvent
was removed under vacuum, and the residue was purified by
chromatography on silica gel (10% ethyl acetate/petroleum) to
afford 3-[1-(2-hydroxy-ethyl)-5-nitro-1H-indol,
2-yl]-3-methyl-butyric acid ethyl ester (270 mg, 60%). .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 8.45 (d, J=2.1 Hz, 1H), 8.05 (dd,
J=2.1, 9.0 Hz, 1H), 6.36 (d, J=9.0 Hz, 1H), 6.48 (s, 1H), 4.46 (t,
J=6.6 Hz, 2H), 4.00-3.91 (m, 4H), 2.76 (s, 2H), 1.61 (s, 6H), 0.99
(t, J=7.2 Hz, 1H), 0.85 (s, 9H), 0.03 (s, 6H).
##STR00636##
3-[1-(2-Hydroxy-ethyl)-5-nitro-1H-indol-2-yl]-3-methyl-butan-1-ol
[0459] To a solution of
3-[1-(2-hydroxyethyl)-5-nitro-1H-indol-2-yl]-3-methyl-butyric acid
ethyl ester (700 rag, 2.1 mmol) in THF (25 mL) was added DIBAL-H
(1.0 M, 4.2 mL, 4.2 mmol) at -78.degree. C. The mixture was stirred
at room temperature for 1 h. Water (2 mL) was added and the
resulting mixture was extracted with EtOAc (15 mL.times.3). The
combined organic layers were dried over anhydrous Na.sub.2SO.sub.4
and evaporated under vacuum. The residue was purified by
chromatography on silica gel (15% ethyl acetate/petroleum) to
afford
3-[1-(2-hydroxy-ethyl)-5-nitro-1H-indol-2-yl]-3-methyl-butan-1-ol
(300 mg, 49%). .sup.1H NMR (300 MHz, d-DMSO) .delta. 8.42 (d, J=1.5
Hz, 1H), 7.95 (dd, J=1.2, 8.7 Hz, 1H), 6.36 (d, J=9.3 Hz, 1H), 6.50
(s, 1H), 5.25 (br s, 1H), 4.46-4.42 (m, 4H), 3.69-3.66 (m, 2H),
3.24-3.21 (m, 2H), 1.42 (s, 6H).
##STR00637##
3-[5-Amino-1-(2-hydroxy-ethyl)-1H-indol-2-yl]-3-methyl-butan-1-ol
[0460] A solution of
3-[1-(2-hydroxy-ethyl)-5-nitro-1H-indol-2-yl]-3-methyl-butan-1-ol
(300 rag, 1.03 mmol) and Raney Nickel (200 mg) in CH.sub.3OH (30
mL) was stirred for 5 h at room temperature under a H.sub.2
atmosphere. The catalyst was filtered through a celite pad and the
filtrate was evaporated under vacuum to give a residue, which was
purified by preparative TLC to afford
3-[5-amino-1-(2-hydroxy-ethyl)-1H-indol-2-yl]-3-methyl-butan-1-ol
(70 rag, 26%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.07 (d,
J=8.7 Hz, 1H), 6.83 (d, J=2.1 Hz, 1H), 6.62 (dd, J=2.1, 8.4 Hz,
1H), 6.15 (s, 1H), 4.47 (t, J=5.4 Hz, 2H), 4.07 (t, J=5.4 Hz, 2H),
3.68 (t, J=5.7 Hz, 2H), 2.16 (t, J=5.7 Hz, 2H), 4.00-3.91 (m, 4H),
2.76 (s, 2H), 1.61 (s, 6H), 1.42 (s, 6H).
Example 48
tert-Butyl 2-(5-amino-1H-indol-2-yl)piperidine-1-carboxylate
##STR00638##
[0461] 2-(Piperidin-2-yl)-1H-indol-5-amine
[0462] 5-Nitro-2-(pyridin-2-yl)-]H-indole (1.0 g, 4.2 mmol) was
added to HCl/MeOH (2 M, 50 mL). The reaction mixture was stirred at
room temperature for 1 h and the solvent was evaporated under
vacuum. PtO.sub.2 (200 mg) was added to a solution of the residue
in MeOH (50 mL) and the reaction mixture was stirred under hydrogen
atmosphere (1 atm) at room temperature for 2 h. The catalyst was
filtered through a celite pad and the solvent was evaporated under
vacuum to afford 2-(piperidin-2-yl)-1H-indol-5-amine (1.0 g), which
was directly used in the next step.
##STR00639##
tert-Butyl 2-(5-amino-1H-indol-2-yl)piperidine-1-carboxylate
[0463] To a solution of 2-(piperidin-2-yl)-1H-indol-5-amine (1.0 g)
in Et.sub.3N (25 mL) and THF (25 mL) was added Boc.sub.2O (640 mg,
2.9 mmol). The reaction mixture was stirred at room temperature
overnight. The mixture was diluted with water and extracted with
dichloromethane (3.times.25 mL). The combined organic layers were
washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuum. The residue was purified by chromatography
on silica gel (5-10% ethyl acetate/petroleum ether) followed by
preparative HPLC to afford tert-butyl
2-(S-amino-1H-indol-2-yl)piperidine-1-carboxylate (15 mg, 1% over 2
steps). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.82 (s, 1H),
7.58 (s, 1H), 7.22 (d, J=8.8 Hz, 1H), 7.02 (d, J=1.6, 8.0 Hz, 1H),
6.42 (s, 1H), 6.25 (s, 1H), 3.91-3.88 (m, 1H), 3.12-3.10 (m, 1H),
2.81-2.76 (m, 1H), 2.06-1.97 (m, 4H), 1.70-1.58 (m, 2H), 1.53 (s,
9H).
Example 49
6-amino-1H-indole-2-carbonitrile
##STR00640##
[0464] (3-Nitrophenyl)hydrazine hydrochloride
[0465] 3-Nitroaniline (28 g, 0.20 mol) was dissolved in a mixture
of H.sub.2O (40 mL) and 37% HCl (40 mL). A solution of NaNO.sub.2
(14 g, 0.20 mol) in H.sub.2O (60 mL) was added to the mixture at
0.degree. C., and then a solution of SnCl.sub.2.H.sub.2O (140 g,
0.60 mol) in 37% HCl (100 mL) was added. After stirring at
0.degree. C. for 0.5 h, the insoluble material was isolated by
filtration and was washed with water to give
(3-nitrophenyl)hydrazine hydrochloride (28 g, 73%).
##STR00641##
(E)-Ethyl 2-(2-(3-nitrophenyl)hydrazono)propanoate
[0466] (3-Nitrophenyl)hydrazine hydrochloride (30 g, 0.16 mol) and
2-oxo-propionic acid ethyl ester (22 g, 0.19 mol) were dissolved in
ethanol (300 mL). The mixture was stirred at room temperature for 4
h before the solvent was evaporated under reduced pressure to give
(E)-ethyl 2-(2-(3-nitrophenyl)hydrazono)propanoate, which was used
directly in the next step.
##STR00642##
Ethyl 4-nitro-1H-indole-2-carboxylate and ethyl
6-nitro-1H-indole-2-carboxylate
[0467] (E)-Ethyl 2-(2-(3-nitrophenyl)hydrazono)propanoate was
dissolved in toluene (300 mL) and PPA (30 g) was added. The mixture
was heated at reflux overnight and then was cooled to room
temperature. The solvent was decanted and evaporated to obtain a
crude mixture that was taken on to the next step without
purification (15 g, 40%).
##STR00643##
4-Nitro-1H-indole-2-carboxylic acid and
6-nitro-1H-indole-2-carboxylic acid
[0468] A mixture of ethyl 6-nitro-1H-indole-2-carboxylate (0.5 g)
and 10% NaOH (20 mL) was heated at reflux overnight and then was
cooled to room temperature. The mixture was extracted with ether
and the aqueous phase was acidified with HCl to pH 1.about.2. The
insoluble solid was isolated by filtration to give a crude mixture
that was taken on to the next step without purification (0.3 g,
68%).
##STR00644##
4-Nitro-1H-indole-2-carboxamide and
6-nitro-1H-indole-2-carboxamide
[0469] A mixture of 6-nitro-1H-indole-2-carboxylic acid (12 g, 58
mmol) and SOCl.sub.2 (50 mL, 64 mmol) in benzene (150 mL) was
heated at reflux for 2 h. The benzene and excess SOCl.sub.2 was
removed under reduced pressure. The residue was dissolved in
anhydrous CH.sub.2Cl.sub.2 (250 mL) and NH.sub.3.H.sub.2O (22 g,
0.32 mol) was added dropwise at 0.degree. C. The mixture was
stirred at room temperature for 1 h. The insoluble solid was
isolated by filtration to obtain crude mixture (9.0 g, 68%), which
was used directly in the next step.
##STR00645##
4-Nitro-1H-indole-2-carbonitrile and
6-nitro-1H-indole-2-carbonitrile
[0470] 6-Nitro-1H-indole-2-carboxamide (5.0 g, 24 mmol) was
dissolved in CH.sub.2Cl.sub.2 (200 mL). Et.sub.3N (24 g, 0.24 mol)
and (CF.sub.3CO).sub.2O (51 g, 0.24 mol) were added dropwise to the
mixture at room temperature. The mixture was continued to stir for
1 h and was then poured into water (100 mL). The organic layer was
separated and the aqueous layer was extracted with EtOAc (100
mL.times.3). The combined organic layers were dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to obtain crude product which was purified by column chromatography
on silica gel to give a impure sample of
4-nitro-1H-indole-2-carbonitrile (2.5 g, 55%).
##STR00646##
6-Amino-1H-indole-2-carbonitrile
[0471] A mixture of 6-nitro-1H-indole-2-carbonitrile (2.5 g, 13
mmol) and Raney Nickel (500 mg) in EtOH (50 mL) was stirred at room
temperature under H.sub.2 (1 atm) for 1 h. Raney Nickel was removed
via filtration and the filtrate was evaporated under reduced
pressure to give a residue, which was purified by column
chromatograpy on silica get to give
6-amino-1H-indole-2-carbonitrile (1.0 g, 49%). .sup.1H NMR
(DMSO-d.sub.6) .delta. 12.75 (br s, 1H), 7.82 (d, J=8 Hz, 1H), 7.57
(s, 1H), 7.42 (s, 1H), 7.15 (d, J=8 Hz, 1H); MS (ESI) m/e
(M+H.sup.+) 158.2.
Example 50
6-Amino-1H-indole-3-carbonitrile
##STR00647##
[0472] 6-Nitro-1H-indole-3-carbonitrile
[0473] To a solution of 6-nitroindole (4.9 g 30 mmol) in DMF (24
mL) and CH.sub.3CN (240 mL) was added dropwise a solution of
ClSO.sub.2NCO (5.0 mL) in CH.sub.3CN (39 mL) at 0.degree. C. After
addition, the reaction was allowed to warm to room temperature and
was stirred for 2 h. The mixture was then poured into ice-water and
basified with sat. NaHCO.sub.3 solution to pH 7-8. The mixture was
extracted with ethyl acetate. The organics were washed with brine,
dried over Na.sub.2SO.sub.4 and concentrated to give
6-nitro-1H-indole-3-carbonitrile (4.6 g, 82%).
##STR00648##
6-Amino-1H-Indole-3-carbonitrile
[0474] A suspension of 6-nitro-1H-indole-3-carbonitrile (4.6 g, 25
mmol) and 10% Pd--C (0.46 g) in EtOH (50 mL) was stirred under Hz
(1 atm) at room temperature overnight. After filtration, the
filtrate was concentrated and the residue was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate=3/1) to
give 6-amino-1H-indole-3-carbonitrile (1.0 g, 98%) as a pink solid.
.sup.1H NMR (DMSO-ds) .delta. 11.51 (s, 1H), 7.84 (d, J=2.4 Hz,
1H), 7.22 (d, J=8.4 Hz, 1H), 6.62 (s, 1H), 6.56 (d, J=8.4 Hz, 1H),
5.0 (s, 2H); MS (ESI) m/e (M+H.sup.+) 157.1.
Example 51
2-tert-Butyl-1H-indol-6-amine
##STR00649##
[0475] N-o-Tolylpivalamide
[0476] To a solution of o-tolylamine (21 g, 0.20 mol) and Et.sub.3N
(22 g, 0.22 mol) in CH.sub.2Cl.sub.2 was added
2,2-dimethyl-propionyl chloride (25 g, 0.21 mol) at 10.degree. C.
After addition, the mixture was stirred overnight at room
temperature. The mixture was washed with aq. HCl (5%, 80 mL),
saturated aq. NaHCO.sub.3 and brine. The organic layer was dried
over Na.sub.2SO.sub.4 and concentrated under vacuum to give
N-o-tolylpivalamide (35 g, 91%). .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 7.88 (d, J=7.2 Hz, 1H), 7.15-7.25 (m, 2H), 7.05 (t, J=7.2
Hz, 1H), 2.26 (s, 3H), 1.34 (s, 9H).
##STR00650##
2-tert-Butyl-1H-indole
[0477] To a solution of N-o-tolylpivalamide (30.0 g, 159 mmol) in
dry THF (100 mL) was added dropwise n-BuLi (2.5 M in hexane, 190
mL) at 15.degree. C. After addition, the mixture was stirred
overnight at 15.degree. C. The mixture was cooled in an ice-water
bath and treated with saturated NH.sub.4Cl. The organic layer was
separated and the aqueous layer was extracted with ethyl acetate.
The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered, and concentrated in vacuum. The residue
was purified by column chromatography on silica gel to give
2-tert-butyl-1H-indole (24 g, 88%). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.99 (br. s, 1H), 7.54 (d, J=7.2 Hz, 1H), 7.05
(d, J=7.8 Hz, 1H), 7.06-7.13 (m, 2H), 6.26 (s, 1H), 1.39 (s,
9H).
##STR00651##
2-tert-Butylindoline
[0478] To a solution of 2-tert-butyl-1H-indole (10 g, 48 mmol) in
AcOH (40 mL) was added NaBH.sub.4 at 10.degree. C. The mixture was
stirred for 20 minutes at 10.degree. C. before being treated
dropwise with H.sub.2O under ice cooling. The mixture was extracted
with ethyl acetate. The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under vacuum
to give 2-tert-butylindoline (9.8 g), which was used directly in
the next step.
##STR00652##
2-tert-butyl-6-nitroindoline and 2-tert-butyl-5-nitro-1H-indole
[0479] To a solution of 2-tert-butylindoline (9.7 g) in
H.sub.2SO.sub.4 (98%, 80 mL) was slowly added KNO.sub.3 (5.6 g, 56
mmol) at 0.degree. C. After addition, the reaction mixture was
stirred at room temperature for 1 h. The mixture was carefully
poured into cracked ice, basified with Na.sub.2CO.sub.3 to pH 8 and
extracted with ethyl acetate. The combined extracts were washed
with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated
under vacuum. The residue was purified by column chromatography to
give 2-tert-butyl-6-nitroindoline (4.0 g, 31% over two steps).
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.52 (dd, J=1.8, 8.1 Hz,
1H), 7.30 (s, 1H), 7.08 (d, J=7.8 Hz, 1H), 3.76 (t, J=9.6 Hz, 1H),
2.98-3.07 (m, 1H), 2.82-2.91 (m, 1H), 0.91 (s, 9H).
##STR00653##
2-tert-Butyl-6-nitro-1H-indole
[0480] To a solution of 2-tert-butyl-6-nitroindoline (2.0 g, 9.1
mmol) in 1,4-dioxane (20 mL) was added DDQ (6.9 g, 30 mmol) at room
temperature. The mixture was heated at reflux for 2.5 h before
being filtered and concentrated under vacuum. The residue was
purified by column chromatography to give
2-tert-butyl-6-nitro-1H-indole (1.6 g, 80%). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.30 (br. s, 1H), 8.29 (s, 1H), 8.00 (dd,
J=2.1, 8.7 Hz, 1H), 7.53 (d, J=9.3 Hz, 1H), 6.38 (s, 1H), 1.43 (s,
9H).
##STR00654##
2-tert-Butyl-1H-indol-6-amine
[0481] To a solution of 2-tert-butyl-6-nitro-1H-indole (1.3 g, 6.0
mmol) in MeOH (10 mL) was added Raney Nickel (0.2 g). The mixture
was hydrogenated under 1 atm of hydrogen at room temperature for 3
h. The reaction mixture was filtered and the filtrate was
concentrated. The residue was washed with petroleum ether to give
2-tert-butyl-1H-indol-6-amine (1.0 g, 89%). .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.19 (s, 1H), 6.99 (d, J=8.1 Hz, 1H), 6.46
(s, 1H), 6.25 (dd, J=1.8, 8.1 Hz, 1H), 5.79 (d, J=1.8 Hz, 1H), 4.52
(s, 2H), 1.24 (s, 9H); MS (ESI) m/e (M+H.sup.+) 189.1.
Example 52
3-tert-Butyl-1H-indol-6-amine
##STR00655##
[0482] 3-tert-Butyl-6-nitro-1H-indole
[0483] To a mixture of 6-nitroindole (1.0 g, 6.2 mmol), zinc
triflate (2.1 g, 5.7 mmol), and TBAI (1.7 g, 5.2 mmol) in anhydrous
toluene (1 mL) was added DIEA (1.5 g, 11 mmol) at room temperature
under nitrogen. The reaction mixture was stirred for 10 min at
120.degree. C., followed by the addition of r-butyl bromide (0.71
g, 5.2 mmol). The resulting mixture was stirred for 45 min at
120.degree. C. The solid was filtered off and the filtrate was
concentrated to dryness. The residue was purified by column
chromatography on silica gel (petroleum ether/ethyl acetate=20:1)
to give 3-tert-butyl-6-nitro-1H-indole (0.25 g, 19%) as a yellow
solid. .sup.1H-NMR (CDCl.sub.3) .delta. 8.32 (d, J=2.1 Hz, 1H),
8.00 (dd, J=2.1, 14.4 Hz, 1H), 7.85 (d, J=8.7 Hz, 1H), 7.25 (s,
1H), 1.46 (s, 9H).
##STR00656##
3-tert-Butyl-1H-indol-6-amine
[0484] A suspension of 3-tert-butyl-6-nitro-1H-indole (3.0 g, 14
mmol) and Raney Nickel (0.5 g) was hydrogenated under H.sub.2 (1
atm) at room temperature for 3 h. The catalyst was filtered off and
the filtrate was concentrated to dryness. The residue was purified
by column on silica gel (petroleum ether/ethyl acetate=4:1) to give
3-tert-butyl-1H-indol-6-amine (2.0 g, 77%) as a gray solid.
.sup.1HNMR (CDCl.sub.3) .delta. 7.58 (m, 2H), 6.73 (d, J=1.2 Hz,
1H), 6.66 (s, 1H), 6.57 (dd, J=0.8, 8.6 Hz, 1H), 3.60 (br, 2H),
1.42 (s, 9H).
Example 53
5-(Trifluoremethyl)-1H-indol-6-amine
##STR00657##
[0485] 1-Methyl-2,4-dinitro-5-(trifluoromethyl)benzene
[0486] To a mixture of HNO.sub.3 (98%, 30 mL) and H.sub.2SO.sub.4
(98%, 30 mL) was added dropwise 1-methyl-3-trifluoromethyl-benzene
(10 g, 63 mmol) at 0.degree. C. After addition, the mixture was
stirred at rt for 30 min and was then poured into ice-water. The
precipitate was filtered and washed with water to give
1-methyl-2,4-dinitro-5-trifluoromethyl-benzene (2.0 g, 13%).
##STR00658##
(E-2-(2,4-Dinitro-5-(trifluoromethyl)phenyl)-N,N-dimethylethenamine
[0487] A mixture of 1-methyl-2,4-dinitro-5-trifluoromethyl-benzene
(2.0 g, 8.0 mmol) and DMA (1.0 g, 8.2 mmol) in DMF (20 mL) was
stirred at 100.degree. C. for 30 min. The mixture was poured into
ice-water and stirred for 1 h. The precipitate was filtered and
washed with water to give
(E)-2-(2,4-dinitro-5-(trifluoromethyl)phenyl)-N,N-dimethylethenamine
(2.1 g, 86%).
##STR00659##
5-(Trifluoromethyl)-1H-indol-6-amine
[0488] A suspension of
(E)-2-(2,4-dinitro-5-(trifluoromethyl)phenyl)-N,N-dimethylethenamine
(2.1 g, 6.9 mmol) and Raney Nickel (1 g) in ethanol (80 mL) was
stirred under H.sub.2 (1 atm) at room temperature for 5 h. The
catalyst was filtered off and the filtrate was concentrated to
dryness. The residue was purified by column on silica gel to give
5-(trifluoromethyl)-1H-indol-6-amine (200 mg, 14%). .sup.1H NMR
(DMSO-ds) .delta. 10.79 (br s, 1H), 7.55 (s, 1H), 7.12 (s, 1H),
6.78 (s, 1H), 6.27 (s, 1H), 4.92 (s, 2H); MS (ESI) m/e (M+H.sup.+):
200.8.
Example 54
5-Ethyl-1H-indol-6-amine
##STR00660## ##STR00661##
[0489] 1-(Phenylsulfonyl)indoline
[0490] To a mixture of DMAP (1.5 g), benzenesulfonyl chloride (24.0
g, 136 mmol) and indoline (14.7 g, 124 mmol) in CH.sub.2Cl.sub.2
(200 mL) was added dropwise Et.sub.3N (19.0 g, 186 mmol) at
0.degree. C. The mixture was stirred at room temperature overnight.
The organic layer was washed with water (2.times.), dried over
Na.sub.2SO.sub.4 and concentrated to dryness under reduced pressure
to obtain 1-(phenylsulfonyl)indoline (30.9 g, 96%).
##STR00662##
1-(1-(Phenylsulfonyl)indolin-5-yl)ethanone
[0491] To a suspension of AlCl.sub.3 (144 g, 1.08 mol) in
CH.sub.2Cl.sub.2 (1070 mL) was added acetic anhydride (54 mL). The
mixture was stirred for 15 minutes before a solution of
1-(phenylsulfonyl)indoline (46.9 g, 0.180 mol) in CH.sub.2Cl.sub.2
(1070 mL) was added dropwise. The mixture was stirred for 5 h and
was quenched by the slow addition of crushed ice. The organic layer
was separated and the aqueous layer was extracted with
CH.sub.2Cl.sub.2. The combined organics were washed with saturated
aqueous NaHCO.sub.3 and brine, dried over Na.sub.2SO.sub.4, and
concentrated under vacuum to obtain
1-(1-(phenylsulfonyl)indolin-5-yl)ethanone (42.6 g).
##STR00663##
5-Ethyl-1-(phenylsulfonyl)indoline
[0492] To TFA (1600 mL) at 0.degree. C. was added sodium
borohydride (64.0 g, 1.69 mol) over 1 h. To this mixture was added
dropwise a solution of 1-(1-(phenylsulfonyl)indolin-5-yl)ethanone
(40.0 g, 0.133 mol) in TFA (700 mL) over 1 h. The mixture was then
stirred overnight at 25.degree. C. After dilution with H.sub.2O
(1600 mL), the mixture was made basic by the addition of sodium
hydroxide pellets at 0.degree. C. The organic layer was separated
and the aqueous layer was extracted with CH.sub.2Cl.sub.2. The
combined organic layers were washed with brine, dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure. The
residue was purified by silica column to give
5-ethyl-1-(phenylsulfonyl)indoline (16.2 g, 47% over two
steps).
##STR00664##
5-Ethylindoline
[0493] A mixture of 5-ethyl-1-(phenylsulfonyl)indoline (15 g, 0.050
mol) in HBr (48%, 162 mL) was heated at reflux for 6 h. The mixture
was basified with sat. NaOH to pH 9 and then it was extracted with
ethyl acetate. The organic layer was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated under reduced pressure. The
residue was purified by silica column to give 5-ethylindoline (2.5
g, 32%).
##STR00665##
5-Ethyl-6-nitroindoline
[0494] To a solution of 5-ethylindoline (2.5 g, 17 mmol) in
H.sub.2SO.sub.4 (98%, 20 mL) was slowly added KNO.sub.3 (1.7 g, 17
mmol) at 0.degree. C. The mixture was stirred at 0-10.degree. C.
for 10 minutes. The mixture was then carefully poured into ice,
basified with NaOH solution to pH 9, and extracted with ethyl
acetate. The combined extracts were washed with brine, dried over
Na.sub.2SO.sub.4 and concentrated to dryness. The residue was
purified by silica column to give 5-ethyl-6-nitroindoline (1.9 g,
58%).
##STR00666##
5-Ethyl-6-nitro-1H-indole
[0495] To a solution of 5-ethyl-6-nitroindoline (1.9 g, 9.9 mmol)
in CH.sub.2Cl.sub.2 (30 mL) was added MnO.sub.2 (4.0 g, 46 mmol).
The mixture was stirred at ambient temperature for 8 h. The solid
was filtered off and the filtrate was concentrated to dryness to
give 5-ethyl-6-nitro-1H-indole (1.9 g).
##STR00667##
5-Ethyl-1H-indol-6-amine
[0496] A suspension of 5-ethyl-6-nitro-1H-indole (1.9 g, 10 mmol)
and Raney Nickel (1 g) was hydrogenated under H.sub.2 (1 atm) at
room temperature for 2 h. The catalyst was filtered off and the
filtrate was concentrated to dryness. The residue was purified by
silica gel column to give 5-ethyl-1H-indol-6-amine (760 mg, 48%
over two steps). .sup.1H NMR (CDCl.sub.3) .delta. 7.90 (br s, 1H),
7.41 (s, 1H), 7.00 (s, 1H), 6.78 (s, 2H), 6.39 (s, 1H), 3.39 (br s,
2H), 2.63 (q, J=7.2 Hz, 2H), 1.29 (t, J=6.9 Hz, 3H); MS (ESI) m/e
(M+H.sup.+) 161.1.
Example 55
Ethyl 6-amino-1H-indole-4-carboxylate
##STR00668##
[0497] 2-Methyl-3,5-dinitrobenzoic acid
[0498] To a mixture of HNO.sub.3 (95%, 80 mL) and H.sub.2SO.sub.4
(98%, 80 mL) was slowly added 2-methylbenzic acid (50 g, 0.37 mol)
at 0.degree. C. After addition, the reaction mixture was stirred
below 30.degree. C. for 1.5 h. The mixture then was poured into
ice-water and stirred for 15 min. The precipitate was filtered and
washed with water to give 2-methyl-3,5-dinitrobenzoic acid (70 g,
84%).
##STR00669##
Ethyl 2-methyl-3,5-dinitrobenzoate
[0499] A mixture of 2-methyl-3,5-dinitrobenzoic acid (50 g, 0.22
mol) in SOCl.sub.2 (80 mL) was heated at reflux for 4 h and then
was concentrated to dryness. The residue was dissolved in
CH.sub.2Cl.sub.2 (50 mL), to which EtOH (80 mL) was added and the
mixture was stirred at room temperature for 1 h. The mixture was
poured into ice-water and extracted with EtOAc (3.times.100 mL).
The combined extracts were washed sat. Na.sub.2CO.sub.3 (80 mL),
water (2.times.100 mL) and brine (100 mL), dried over
Na.sub.2SO.sub.4 and concentrated to dryness to give ethyl
2-methyl-3,5-dinitrobenzoate (50 g, 88%)
##STR00670##
(E)-Ethyl 2-(2-(dimethylamino)vinyl)-3,5-dinitrobenzoate
[0500] A mixture of ethyl 2-methyl-3,5-dinitrobenzoate (35 g, 0.14
mol) and DMA (32 g, 0.27 mol) in DMF (200 mL) was heated at
100.degree. C. for 5 h. The mixture was poured into ice-water and
the precipitated solid was filtered and washed with water to give
(E)-ethyl 2-(2-(dimethylamino)vinyl)-3,5-dinitrobenzoate (11 g,
48%)
##STR00671##
Ethyl 6-amino-1H-indole-4-carboxylate
[0501] A mixture of (E)-ethyl
2-(2-(dimethylamino)vinyl)-3,5-dinitrobenzoate (11 g, 0.037 mol)
and SnCl.sub.2 (83 g, 0.37 mol) in ethanol was heated at reflux for
4 h. The mixture was concentrated to dryness and the residue was
poured into water and basified using sat. aq. Na.sub.2CO.sub.3 to
pH 8. The precipitated solid was filtered and the filtrate was
extracted with ethyl acetate (3.times.100 mL). The combined
extracts were washed with water (2.times.100 mL) and brine (150
mL), dried over Na.sub.2SO.sub.4 and concentrated to dryness. The
residue was purified by column on silica gel to give ethyl
6-amino-1H-indole-4-carboxylate (3.0 g, 40%). .sup.1HNMR
(DMSO-d.sub.6) .delta. 10.76 (br s, 1H), 7.11-7.14 (m, 2H),
6.81-6.82 (m, 1H), 6.67-6.68 (m, 1H), 4.94 (br s, 2H), 4.32-4.25
(q, J=7.2 Hz, 2H), 1.35-1.31 (t, J=7.2, 3 H); MS (ESI) m/e
(M+H.sup.+) 205.0.
Example 56
5-Fluoro-1H-indol-6-amine
##STR00672##
[0502] 1-Fluoro-5-methyl-2,4-dinitrobenzene
[0503] To a stirred solution of HNO.sub.3 (60 mL) and
H.sub.2SO.sub.4 (80 mL) was added dropwise 1-fluoro-3-methylbenzene
(28 g, 25 mmol) under ice-cooling at such a rate that the
temperature did not rise above 35.degree. C. The mixture was
allowed to stir for 30 min at rt and was then poured into ice water
(500 mL). The resulting precipitate (a mixture of
1-fluoro-5-methyl-2,4-dinitrobenzene and
1-fluoro-3-methyl-2,4-dinitrobenzene, 32 g, ca. 7:3 ratio) was
collected by filtration and purified by recrystallization from 50
mL isopropyl ether to give pure
1-fluoro-5-methyl-2,4-dinitro-benzene as a white solid (18 g,
36%).
##STR00673##
(E)-2-(5-Fluoro-2,4-dinitrophenyl)-N,N-dimethylethenamine
[0504] A mixture of 1-fluoro-5-methyl-2,4-dinitro-benzene (10 g, 50
mmol), DMA (12 g, 100 mmol) and DMF (50 mL) was heated at
100.degree. C. for 4 h. The solution was cooled and poured into
water. The precipitated red solid was collected, washed with water,
and dried to give
(E)-2-(5-fluoro-24-dinitrophenyl)-N,N-dimethylethenamine (8.0 g,
63%).
##STR00674##
5-Fluoro-1H-indol-6-amine
[0505] A suspension of
(E)-2-(5-fluoro-2,4-dinitrophenyl)-N,N-dimethylethenamine (8.0 g,
31 mmol) and Raney Nickel (8 g) in EtOH (80 mL) was stirred under
H.sub.2 (40 psi) at room temperature for 1 h. After filtration, the
filtrate was concentrated and the residue was purified by column
chromatography (petroleum ether/ethyl acetate=5/1) to give
5-fluoro-1H-indol-6-amine (1.0 g, 16%) as a brown solid. .sup.1HNMR
(DMSO-d.sub.6) .delta. 10.56 (br s, 1H), 7.07 (d, J=12 Hz, 1H),
7.02 (m, 1H), 6.71 (d, J=8 Hz, 1H), 6.17 (s, 1H), 3.91 (br s, 2H);
MS (ESI) m/e (M+H.sup.+) 150.1.
Example 57
5-Chloro-1H-indol-6-amine
##STR00675##
[0506] 1-Chloro-5-methyl-2,4-dinitrobenzene
[0507] To a stirred solution of HNO.sub.3 (55 mL) and
H.sub.2SO.sub.4 (79 mL) was added dropwise 1-chloro-3-methylbenzene
(25.3 g, 200 mmol) under ice-cooling at such a rate that the
temperature did not rise above 35.degree. C. The mixture was
allowed to stir for 30 min at ambient temperature and was then
poured into ice water (500 mL). The resulting precipitate was
collected by filtration and purified by recrystallization to give
1-chloro-5-methyl-2,4-dinitrobenzene (26 g, 60%).
##STR00676##
(E)-2-(5-Chloro-2,4-dinitrophenyl)-N,N-dimethylethenamine
[0508] A mixture of 1-chloro-5-methyl-2,4-dinitro-benzene (11.6 g,
50.0 mmol), DMA (11.9 g, 100 mmol) in DMF (50 mL) was heated at
100.degree. C. for 4 h. The solution was cooled and poured into
water. The precipitated red solid was collected by filtration,
washed with water, and dried to give
(E)-2-(5-chloro-2,4-dinitrophenyl)-N,N-dimethylethenamine (9.84 g,
72%).
##STR00677##
5-Chloro-1H-indol-6-amine
[0509] A suspension of
(E)-2-(5-chloro-2,4-dinitrophenyl)-N,N-dimethylethenamine (9.8 g,
36 mmol) and Raney Nickel (9.8 g) in EtOH (140 mL) was stirred
under H.sub.2 (1 atm) at room temperature for 4 h. After
filtration, the filtrate was concentrated and the residue was
purified by column chromatograph (petroleum ether/ethyl
acetate=10:1) to give 5-chloro-1H-indol-6-amine (0.97 g, 16%) as a
gray powder. .sup.1HNMR (CDCl.sub.3) .delta. 7.85 (br s, 1H), 7.52
(s, 1H), 7.03 (s, 1H), 6.79 (s, 1H), 6.34 (s, 1H), 3.91 (br s, 1H);
MS (ESI) m/e (M+H.sup.+) 166.0.
Example 58
Ethyl 6-amino-1H-indole-7-carboxylate
##STR00678##
[0510] 3-Methyl-2,6-dinitrobenzoic acid
[0511] To a mixture of HNO.sub.3 (95%, 80 mL) and H.sub.2SO.sub.4
(98%, 80 mL) was slowly added 3-methylbenzic acid (50 g, 0.37 mol)
at 0.degree. C. After addition, the mixture was stirred below
30.degree. C. for 1.5 hours. The mixture was then poured into
ice-water and stirred for 15 min. The precipitate solid was
filtered and washed with water to give a mixture of
3-methyl-2,6-dinitro-benzoic acid and 5-methyl-2,4-dinitrobenzoic
acid (70 g, 84%). To a solution of this mixture (70 g, 0.31 mol) in
EtOH (150 mL) was added dropwise SOCl.sub.2 (54 g, 0.45 mol). The
mixture was heated at reflux for 2 h before being concentrated to
dryness under reduced pressure. The residue was partitioned between
EtOAc (100 mL) and aq. Na.sub.2CO.sub.3 (10%, 120 mL). The organic
layer was washed with brine (50 mL), dried over Na.sub.2SO.sub.4,
and concentrated to dryness to obtain ethyl
5-methyl-2,4-dinitrobenzoate (20 g), which was placed aside. The
aqueous layer was acidified by HCl to pH 2-3 and the precipitated
solid was filtered, washed with water, and dried in air to give
3-methyl-2,6-dinitrobenzoic acid (39 g, 47%).
##STR00679##
Ethyl 3-methyl-2,6-dinitrobenzoate
[0512] A mixture of 3-methyl-2,6-dinitrobenzoic acid (39 g, 0.15
mol) and SOCl.sub.2 (80 mL) was heated at reflux 4 h. The excess
SOCl.sub.2 was evaporated off under reduced pressure and the
residue was added dropwise to a solution of EtOH (100 mL) and
Et.sub.3N (50 mL). The mixture was stirred at 20.degree. C. for 1 h
and then concentrated to dryness. The residue was dissolved in
EtOAc (100 mL), washed with Na.sub.2CO.sub.3 (10%, 40 mL.times.2),
water (50 mL.times.2) and brine (50 mL), dried over
Na.sub.2SO.sub.4 and concentrated to give ethyl
3-methyl-2,6-dinitrobenzoate (20 g, 53%).
##STR00680##
(E)-Ethyl 3-(2-(dimethylamino)vinyl)-2,6-dinitrobenzoate
[0513] A mixture of ethyl 3-methyl-2,6-dinitrobenzoate (35 g, 0.14
mol) and DMA (32 g, 0.27 mol) in DMF (200 mL) was heated at
100.degree. C. for 5 h. The mixture was poured into ice water. The
precipitated solid was filtered and washed with water to give
(E)-ethyl 3-(2-(dimethylamino)vinyl)-2,6-dinitrobenzoate (25 g,
58%).
##STR00681##
Ethyl 6-amino-1H-indole-7-carboxylate
[0514] A mixture of (E)-ethyl
3-(2-(dimethylamino)vinyl)-2,6-dinitrobenzoate (30 g, 0.097 mol)
and Raney Nickel (10 g) in EtOH (1000 mL) was hydrogenated at room
temperature under 50 psi for 2 h. The catalyst was filtered off and
the filtrate was concentrated to dryness. The residue was purified
by column on silica gel to give ethyl
6-amino-1H-indole-7-carboxylate as an off-white solid (3.2 g, 16%).
.sup.1H NMR (DMSO-d.sub.6) .delta. 10.38 (s, 1H), 7.42 (d, J=8.7
Hz, 1H), 6.98 (t, J=3.0 Hz, 1H), 6.65 (s, 2H), 6.48 (d, J=8.7 Hz,
1H), 6.27-6.26 (m, 1H), 4.38 (q, J=7.2 Hz, 2H), 1.35 (t, J=7.2 Hz,
3H).
Example 59
Ethyl 6-amino-1H-indole-5-carboxylate
##STR00682##
[0515] (E)-Ethyl 5-(2-(dimethylamino)vinyl)-2,4-dinitrobenzoate
[0516] A mixture of ethyl 5-methyl-2,4-dinitrobenzoate (39 g, 0.15
mol) and DMA (32 g, 0.27 mol) in DMF (200 mL) was heated at
100.degree. C. for 5 h. The mixture was poured into ice water and
the precipitated solid was filtered and washed with water to afford
(E)-ethyl 5-(2-(dimethylamino)vinyl)-2,4-dinitrobenzoate (15 g,
28%).
##STR00683##
Ethyl 6-amino-1H-indole-5-carboxylate
[0517] A mixture of (E)-ethyl
5-(2-(dimethylamino)vinyl)-2,4-dinitrobenzoate (15 g, 0.050 mol)
and Raney Nickel (5 g) in EtOH (500 mL) was hydrogenated at room
temperature under 50 psi of hydrogen for 2 h. The catalyst was
filtered off and the filtrate was concentrated to dryness. The
residue was purified by column on silica gel to give ethyl
6-amino-1H-indole-5-carboxylate (3.0 g, 30%). .sup.1H NMR
(DMSO-d.sub.6) .delta. 10.68 (s, 1H), 7.99 (s, 1H), 7.01-7.06 (m,
1H), 6.62 (s, 1H), 6.27-6.28 (m, 1H), 6.16 (s, 2H), 4.22 (q, J=7.2
Hz, 2H), 1.32-1.27 (t, J=7.2 Hz, 3H).
Example 60
5-tert-Butyl-1H-indol-6-amine
##STR00684##
[0518] 2-tert-Butyl-4-methylphenyl diethyl phosphate
[0519] To a suspension of NaH (60% in mineral oil, 8.4 g, 0.21 mol)
in THF (200 mL) was added dropwise a solution of
2-tert-butyl-4-methylphenol (33 g, 0.20 mol) in THF (100 mL) at
0.degree. C. The mixture was stirred at 0.degree. C. for 15 min and
then phosphorochloridic acid diethyl ester (37 g, 0.21 mol) was
added dropwise at 0.degree. C. After addition, the mixture was
stirred at ambient temperature for 30 min. The reaction was
quenched with sat. NH.sub.4CI (300 mL) and then extracted with
Et.sub.2O (350 mL.times.2). The combined organic layers were washed
with brine, dried over anhydrous Na.sub.2SO.sub.4, and then
evaporated under vacuum to give 2-tert-butyl-4-methylphenyl diethyl
phosphate (contaminated with mineral oil) as a colorless oil (60 g,
.about.100%), which was used directly in the next step.
##STR00685##
1-tert-Butyl-3-methylbenzene
[0520] To NH.sub.3 (liquid, 1000 mL) was added a solution of
2-tert-butyl-4-methylphenyl diethyl phosphate (60 g, crude from
last step, about 0.2 mol) in Et.sub.2O (anhydrous, 500 mL) at
-78.degree. C. under N.sub.2 atmosphere. Lithium metal was added to
the solution in small pieces until the blue color persisted. The
reaction mixture was stirred at -78.degree. C. for 15 min and then
was quenched with sat. NH.sub.4CI until the mixture turned
colorless. Liquid NH.sub.3 was evaporated and the residue was
dissolved in water. The mixture was extracted with Et.sub.2O (400
mL.times.2). The combined organics were dried over Na.sub.2SO.sub.4
and evaporated to give 1-tert-butyl-3-methylbenzene (contaminated
with mineral oil) as a colorless oil (27 g, 91%), which was used
directly in next step.
##STR00686##
1-tert-Butyl-5-methyl-2,4-dinitrobenzene and
1-tert-butyl-3-methyl-2,4-dinitro-benzene
[0521] To HNO.sub.3 (95%, 14 mL) was added H.sub.2SO.sub.4 (98%, 20
mL) at 0.degree. C. and then 1-tert-butyl-3-methylbenzene (7.4 g,
.about.50 mmol, crude from last step) dropwise to the with the
temperature being kept below 30.degree. C. The mixture was stirred
at ambient temperature for 30 min, poured onto crushed ice (100 g),
and extracted with EtOAc (50 mL.times.3). The combined organic
layers were washed with water and brine, before being evaporated to
give a brown oil, which was purified by column chromatography to
give a mixture of 1-tert-butyl-5-methyl-2,4-dinitrobenzene and
I-tert-butyl-3-methyl-2,4-dinitrobenzene (2:1 by NMR) as a yellow
oil (9.0 g 61%).
##STR00687##
(E)-2-(5-tert-Butyl-2,4-dinitrophenyl)-N,N-dimethylethenamine
[0522] A mixture of 1-tert-butyl-5-methyl-2,4-dinitrobenzene and
1-tert-butyl-3-methyl-2,4-dinitrobenzene (9.0 g, 38 mmol, 2:1 by
NMR) and DMA (5.4 g, 45 mmol) in DMF (50 mL) was heated at reflux
for 2 h before being cooled to room temperature. The reaction
mixture was poured into water-ice and extracted with EtOAc (50
mL.times.3). The combined organic layers were washed with water and
brine, before being evaporated to give a brown oil, which was
purified by column to give
(E)-2-(5-tert-butyl-2,4-dinitrophenyl)-N,N-dimethylethen-amine (5.0
g, 68%).
##STR00688##
5-tert-Butyl-1H-indol-6-a mine
[0523] A solution of
(E)-2-(5-tert-butyl-2,4-dinitrophenyl)-N,N-dimethylethen-amine (5.3
g, 18 mmol) and tin (II) chloride dihydrate (37 g, 0.18 mol) in
ethanol (200 mL) was heated at reflux overnight. The mixture was
cooled to room temperature and the solvent was removed under
vacuum. The residual slurry was diluted with water (500 mL) and was
basifed with 10% aq. Na.sub.2CO.sub.3 to pH 8. The resulting
suspension was extracted with ethyl acetate (3.times.100 mL). The
ethyl acetate extract was washed with water and brine, dried over
Na.sub.2SO.sub.4, and concentrated. The residual solid was washed
with CH.sub.2Cl.sub.2 to afford a yellow powder, which was purified
by column chromatography to give 5-tert-butyl-1H-indol-6-amine
(0.40 g, 12%). .sup.1H NMR (DMSO.d.sub.6) .delta. 10.34 (br s, 1H),
7.23 (s, 1H), 6.92 (s, 1H), 6.65 (s, 1H), 6.14 (s, 1H), 4.43 (br s,
2H), 2.48 (s, 9H); MS (ESI) m/e (M+H.sup.4) 189.1.
General Procedure IV
Synthesis of acylaminoindoles
##STR00689##
[0525] One equivalent of the appropriate carboxylic acid and one
equivalent of the appropriate amine were dissolved in
N,N-dimethylformamide (DMF) containing triethylamine (3
equivalents).
O-(7-Azabenzotriazol-1-yl)-N,N,N,N'-tetramethyluronium
hexafluorophosphate (HATU) was added and the solution was allowed
to stir. The crude product was purified by reverse-phase
preparative liquid chromatography to yield the pure product.
Example 61
N-(2-tert-Butyl-1H-indol-5-yl)-1-(4-methoxyphenyl)-cyclopropanecarboxamide
##STR00690##
[0527] 2-tert-Butyl-1H-indol-5-amine (19 mg, 0.10 mmol) and
1-(4-methoxyphenyl)-cyclopropanecarboxylic acid (19 mg, 0.10 mmol)
were dissolved in N,N-dimethylformamide (1.00 mL) containing
triethylamine (28 .mu.L, 0.20 mmol). O-(7-Azabenzotriazol-1-yl);
N,N,N',N'-tetramethyluronium hexafluorophosphate (42 mg, 0.11 mmol)
was added to the mixture and the resulting solution was allowed to
stir for 3 hours. The crude reaction mixture was filtered and
purified by reverse phase HPLC. ESI-MS m/z calc. 362.2, found 363.3
(M+1).sup.+; Retention time 3.48 minutes.
General Procedure V
Synthesis of acylaminoindoles
##STR00691##
[0529] One equivalent of the appropriate carboxylic acid was placed
in an oven-dried flask under nitrogen. A minimum (3 equivalents) of
thionyl chloride and a catalytic amount of and
N,N-dimethylformamide were added and the solution was allowed to
stir for 20 minutes at 60.degree. C. The excess thionyl chloride
was removed under vacuum and the resulting solid was suspended in a
minimum of anhydrous pyridine. This solution was slowly added to a
stirred solution of one equivalent the appropriate amine dissolved
in a minimum of anhydrous pyridine. The resulting mixture was
allowed to stir for 15 hours at 110.degree. C. The mixture was
evaporated to dryness, suspended in dichloromethane, and then
extracted three times with 1N HCl. The organic layer was then dried
over sodium sulfate, evaporated to dryness, and then purified by
column chromatography.
Example 62
Ethyl
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indole-2--
carboxylate (Compd. 28)
##STR00692##
[0531] 1-Benzo[1,3]dioxol-5-yl-cyclopropanecarboxylic acid (2.07 g,
10.0 mmol) was dissolved in thionyl chloride (2.2 mL) under
N.sub.2. N,N-dimethylformamide (0.3 mL) was added and the solution
was allowed to stir for 30 minutes. The excess thionyl chloride was
removed under vacuum and the resulting solid was dissolved in
anhydrous dichloromethane (15 mL) containing triethylamine (2.8 mL,
20.0 mmol). Ethyl 5-amino-1H-indole-2-carboxylate (2.04 g, 10.0
mmol) in 15 mL of anhydrous dichloromethane was slowly added to the
reaction. The resulting solution was allowed to stir for 1 hour.
The reaction mixture was diluted to 50 mL with dichloromethane and
washed three times with 50 mL of 1N HCl, saturated aqueous sodium
bicarbonate, and saturated aqueous sodium chloride. The organic
layer was dried over sodium sulfate and evaporated to dryness to
yield ethyl
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indole-2-carbo-
xylate as a gray solid (3.44 g, 88%). ESI-MS m/z calc. 392.4; found
393.1 (M+1) Retention time 3.17 minutes. .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 11.80 (s, 1H), 8.64 (s, 1H), 7.83 (m, 1H),
7.33-7.26 (m, 2H), 7.07 (m, 1H), 7.02 (m, 1H), 6.96-6.89 (m, 2H),
6.02 (s, 2H), 4.33 (q, J=7.1 Hz, 2H), 1.42-1.39 (m, 2H), 1.33 (t,
J=7.1 Hz, 3H), 1.06-1.03 (m, 21).
Example 63
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)cyclopropanecar-
boxamide
##STR00693##
[0533] 1-Benzo[1,3]dioxol-5-yl-cyclopropanecarboxylic acid (1.09 g,
5.30 mmol) was dissolved in 2 mL of thionyl chloride under
nitrogen. A catalytic amount (0.3 mL) of N,N-dimethylformamide
(DMF) was added and the reaction mixture was stirred for 30
minutes. The excess thionyl chloride was evaporated and the
resulting residue was dissolved in 15 mL of dichloromethane. This
solution was slowly added to a solution of
2-tert-butyl-1H-indol-5-amine (1.0 g, 5.3 mmol) in 10 mL of
dichloromethane containing triethylamine (1.69 mL, 12.1 mmol). The
resulting solution was allowed to stir for 10 minutes. The solvent
was evaporated to dryness and the crude reaction mixture was
purified by silica gel column chromatography using a gradient of
5-50% ethyl acetate in hexanes. The pure fractions were combined
and evaporated to dryness to yield a pale pink powder (1.24 g 62%).
ESI-MS m/z calc. 376.18, found 377.3 (M+1)+. Retention time of 3.47
minutes. .sup.1H NMR (400 MHz, DMSO) .delta. 10.77 (s, 1H), 8.39
(s, 1H), 7.56 (d, J=1.4 Hz, 1H), 7.15 (d, J=8.6 Hz, 1H), 7.05-6.87
(m, 4H), 6.03 (s, 3H), 1.44-1.37 (m, 2H), 1.33 (s, 9H), 1.05-1.00
(m, 2H).
Example 64
1-(Benzo[d][1,3]dioxol-5-yl)-N-(1-methyl-2-(1-methylcyclopropyl)-1H-indol--
5-yl)cyclopropanecarboxamide
##STR00694##
[0535] 1-Methyl-2-(1-methylcyclopropyl)-1H-indol-5-amine (20.0 mg,
0.100 mmol) and 1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic
acid (20.6 mg, 0.100 mmol) were dissolved in N,N-dimethylformamide
(1 mL) containing triethylamine (42.1 .mu.L, 0.300 mmol) and a
magnetic stir bar.
O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (42 mg, 0.11 mmol) was added to the mixture and
the resulting solution was allowed to stir for 6 h at 80.degree. C.
The crude product was then purified by preparative HPLC utilizing a
gradient of 0-99% acetonitrile in water containing 0.05%
trifluoroacetic acid to yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(1-methyl-2-(1-methylcyclopropyl)-1H-
-indol-5-yl)cyclopropanecarboxamide. ESI-MS m/z calc. 388.2, found
389.2 (M+1).sup.+. Retention time of 3.05 minutes.
Example 65
1-(Benzo[d][1,3]dioxol-5-yl)-N-(1,1-dimethyl-2,3-dihydro-1H-pyrrolo[[1,2-a-
]indol-7-yl)cyclopropanecarboxamide
##STR00695##
[0537] 1,1-Dimethyl-2,3-dihydro-1H-pyrrolo[1,2-a]indol-7-amine
(40.0 mg, 0.200 mmol) and
1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid (41.2 mg,
0.200 mmol) were dissolved in N,N-dimethylformamide (1 mL)
containing triethylamine (84.2 .mu.L, 0.600 mmol) and a magnetic
stir bar. O-(7-Azabenzotriazol-1-yl)-N,N,N'N'-tetramethyluronium
hexafluorophosphate (84 mg, 0.22 mmol) was added to the mixture and
the resulting solution was allowed to stir for 5 minutes at room
temperature. The crude product was then purified by preparative
HPLC utilizing a gradient of 0-99% acetonitrile in water containing
0.05% trifluoroacetic acid to yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(1,1-dimethyl-2,3-dihydro-1H-pyrrolo[1,2-a-
]-indol-7-yl)cyclopropanecarboxamide. ESI-MS m/z calc. 388.2, found
389.2 (M+1).sup.+. Retention time of 2.02 minutes. .sup.1H NMR (400
MHz, DMSO-d6) .delta. 8.41 (s, 1H), 7.59 (d, J, 1.8 Hz, 1H), 7.15
(d, J=8.6 Hz, 1H), 7.06-7.02 (m, 2H), 6.96-6.90 (m, 2H), 6.03 (s,
2H), 5.98 (d, J=0.7 Hz, 1H), 4.06 (t, J=6.8 Hz, 2H), 2.35 (t, J=6.8
Hz, 2H), 1.42-1.38 (m, 2H), 1.34 (s, 6H), 1.05-1.01 (m, 2H).
Example 66
Methyl
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-buty-
l-1H-Indole-7-carboxylate
##STR00696##
[0539] 1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarbonyl chloride
(45 mg, 0.20 mmol) and methyl
5-amino-2-tert-butyl-1H-indole-7-carboxylate (49.3 mg, 0.200 mmol)
were dissolved in N,N-dimethylformamide (2 mL) containing a
magnetic stir bar and triethylamine (0.084 mL, 0.60 mmol). The
resulting solution was allowed to stir for 10 minutes at room
temperature. The crude product was then purified by preparative
HPLC using a gradient of 0-99% acetonitrile in water containing
0.05% trifluoroacetic acid to yield methyl
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarbox-amido)-2-tert-butyl-1H--
indole-7-carboxylate. ESI-MS m/z calc. 434.2, found 435.5
(M+1).sup.+. Retention time of 2.12 minutes.
Example 67
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol-
-5-yl)cyclopropanecarboxamide
##STR00697##
[0541] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid (0.075 g,
0.36 mmol) in acetonitrile (1.5 mL) were added HBTU (0.138 g, 0.36
mmol) and Et.sub.3N (152 .mu.L, 1.09 mmol) at room temperature. The
mixture was stirred at room temperature for 10 minutes before a
solution of 2-(5-amino-1H-indol-2-yl)-2-methylpropan-1-ol (0.074 g,
0.36 mmol) in acetonitrile (1.94 mL) was added. After addition, the
reaction mixture was stirred at room temperature for 3 h. The
solvent was evaporated under reduced pressure and the residue was
dissolved in dichloromethane. The organic layer was washed with 1 N
HCl (1.times.3 mL) and saturated aqueous NaHCO.sub.3 (1.times.3
mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered
and evaporated under reduced pressure. The crude material was
purified by column chromatography on silica gel (ethyl
acetate/hexane=1/1) to give
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-(1-hydroxy-2-methylpropan-2-yl)-1H-indo-
l-5-yl)cyclopropanecarboxamide (0.11 g, 75%). .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 10.64 (s, 1H), 8.38 (s. 1H), 7.55 (s, 1H), 7.15
(d, J=8.6 Hz, 1H), 7.04-6.90 (m, 4H), 6.06 (s, 1H), 6.03 (s, 2H),
4.79 (t, J=2.7 Hz, 1H), 3.46 (d, J=0.0 Hz, 2H), 1.41-1.39 (m, 2H),
1.26 (s, 6H), 1.05-1.02 (m, 2H).
Example 67
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2,3,4,9-tetrahydro-1H-carbazol-6-yl)cyclop-
ropanecarboxamide
##STR00698##
[0543] 2,3,4,9-Tetrahydro-1H-carbazol-6-amine (81.8 mg, 0.439 mmol)
and 1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid (90.4
ag, 0.439 mmol) were dissolved in acetonitrile (3 mL) containing
diisopropylethylamine (0.230 mL, 1.32 mmol) and a magnetic stir
bar. O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (183 mg, 0.482 mmol) was added to the mixture
and the resulting solution was allowed to stir for 16 h at
70.degree. C. The solvent was evaporated and the crude product was
then purified on 40 g of silica gel utilizing a gradient of 5-50%
ethyl acetate in hexanes to yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(2,3,4,9-tetrahydro-1H-carbazol-6-yl-
)cyclopropanecarboxamide as a beige powder (0.115 g, 70%) after
drying. ESI-MS m/z calc. 374.2, found 375.3 (M+1).sup.+. Retention
time of 3.43 minutes. .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.52
(s, 1H), 8.39 (s, 1H), 7.46 (d, J=1.8 Hz, 1H), 7.10-6.89*(m, 5H),
6.03 (s, 2H), 2.68-2.65 (m, 2H), 2.56-2.54 (m, 2H), 1.82-1.77 (m,
4H), 1.41-1.34 (m, 2H), 1.04-0.97 (m, 2H).
Example 69
tert-Butyl
4-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-i-
ndol-2-yl)piperidine-1-carboxylate
##STR00699##
[0545] 1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarbonyl chloride
(43 mg, 0.19 mmol) and tert-butyl
4-(5-amino-1H-indol-2-yl)piperidine-1-carboxylate (60 mg, 0.19
mmol) were dissolved in dichloromethane (1 mL) containing a
magnetic stir bar and triethylamine (0.056 mL, 0.40 mmol). The
resulting solution was allowed to stir for two days at room
temperature. The crude product was then evaporated to dryness,
dissolved in a minimum of N,N-dimethylformamide, and then purified
by preparative HPLC using a gradient of 0-99% acetonitrile in water
containing 0.05% trifluoroacetic acid to yield tert-butyl
4-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-2-yl)-
piperidine-1-carboxylate. ESI-MS m/z calc. 503.2, found 504.5
(M+1). Retention time of 1.99 minutes.
Example 70
Ethyl
2-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol--
2-yl)propanoate
##STR00700##
[0546] tert-Butyl
2-(1-ethoxy-1-oxopropan-2-yl)-1H-indole-1-carboxylate
[0547] tert-Butyl 2-(2-ethoxy-2-oxoethyl)-1H-indole-1-carboxylate
(3.0 g, 9.9 mmol) was added to anhydrous THF (29 mL) and cooled to
-78.degree. C. A 0.5M solution of potassium hexamethyldisilazane
(20 mL, 9.9 mmol) was added slowly such that the internal
temperature stayed below -60.degree. C. Stirring was continued for
1 h at -78.degree. C. Methyl iodide (727 .mu.L, 11.7 mmol) was
added to the mixture. The mixture was stirred for 30 minutes at
room temperature. The mixture was quenched with sat. aq. ammonium
chloride and partitioned between water and dichloromethane. The
aqueous phase was extracted with dichloromethane and the combined
organic phases were dried over Na.sub.2SO.sub.4 and evaporated
under reduced pressure. The residue was purified by column
chromatography on silica gel (ethylacetate/hexane=1/9) to give
tert-butyl 2-(1-ethoxy-1-oxopropan-2-yl)-1H-indole-1-carboxylate
(2.8 g, 88%).
##STR00701##
Ethyl 2-(1H-indol-2-yl)propanoate
[0548] tert-Butyl
2-(1-ethoxy-1-oxopropan-2-yl)-1H-indole-1-carboxylate (2.77 g, 8.74
mmol) was dissolved in dichloromethane (25 mL) before TFA (9.8 mL)
was added. The mixture was stirred for 1.5 h at room temperature.
The mixture was evaporated to dryness, taken up in dichloromethane
and washed with sat. aq. sodium bicarbonate, water, and brine. The
product was purified by column chromatography on silica gel (0-20%
EtOAc in hexane) to give ethyl 2-(1H-indol-2-yl)propanoate (0.92 g,
50%).
##STR00702##
Ethyl 2-(5-nitro-1H-indol-2-yl)propanoate
[0549] Ethyl 2-(1H-indol-2-yl)propanoate (0.91 g, 4.2 mmol) was
dissolved in concentrated sulfuric acid (3.9 mL) and cooled to
-10.degree. C. (salt/ice-mixture). A solution of sodium nitrate
(0.36 g, 4.2 mmol) in concentrated sulfuric acid (7.8 mL) was added
dropwise over 35 min. Stirring was continued for another 30 min at
-10.degree. C. The mixture was poured into ice and the product was
extracted with ethyl acetate. The combined organic phases were
washed with a small amount of sat. aq. sodium bicarbonate. The
product was purified by column chromatography on silica gel (5-30%
EtOAc in hexane) to give ethyl 2-(5-nitro-1H-indol-2-yl)propanoate
(0.34 g, 31%).
##STR00703##
Ethyl 2-(5-amino-1H-indol-2-yl)propanoate
[0550] To a solution of ethyl 2-(5-nitro-1H-indol-2-yl)propanoate
(0.10 g, 0.38 mmol) in ethanol (4 mL) was added tin chloride
dihydrate (0.431 g, 1.91 mmol). The mixture was heated in the
microwave at 120.degree. C. for 1 h. The mixture was diluted with
ethyl acetate before water and saturated aqueous NaHCO.sub.3 were
added. The reaction mixture was filtered through a plug of celite
using ethyl acetate. The organic layer was separated from the
aqueous layer. The organic layer was dried over Na.sub.2SO.sub.4,
filtered and evaporated under reduced pressure to give ethyl
2-(5-amino-1H-indol-2-yl)propanoate (0.088 g, 99%).
##STR00704##
Ethyl
2-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-
-2-yl)propanoate
[0551] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid (0.079 g,
0.384 mmol) in acetonitrile (1.5 mL) were added HBTU (0.146 g,
0.384 mmol) and Et.sub.3N (160 .mu.L, 1.15 mmol) at room
temperature. The mixture was allowed to stir at room temperature
for 10 min before a solution of ethyl
2-(5-amino-1H-indol-2-yl)propanoate (0.089 g, 0.384 mmol) in
acetonitrile (2.16 mL) was added. After addition, the reaction
mixture was stirred at room temperature for 2 h. The solvent was
evaporated under reduced pressure and the residue was dissolved in
dichloromethane. The organic layer was washed with 1 N HCl
(1.times.3 mL) and then saturated aqueous NaHCO.sub.3 (1.times.3
mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered
and evaporated under reduced pressure. The crude material was
purified by column chromatography on silica gel (ethyl
acetate/hexane=1/1) to give ethyl
2-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-2-yl)-
propanoate (0.081 g, 50%). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.51 (s, 1H), 7.67 (s, 1H), 7.23-7.19 (m, 2H), 7.04-7.01
(m, 3H), 6.89 (d, J=0.0 Hz, 1H), 6.28 (s, 1H), 6.06 (s, 2H),
4.25-4.17 (m, 2H), 3.91 (q, J=7.2 Hz, 1H), 1.72-1.70 (m, 2H), 1.61
(s, 2H), 1.29 (t, J=7.1 Hz, 4H), 1.13-1.11 (m, 2H).
Example 71
tert-Butyl
2-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarbox-amido
1H-indol-2-yl)-2-methylpropylcarbamate
##STR00705##
[0552] 2-Methyl-2-(S-nitro-1H-indol-2-yl)propanoic acid
[0553] Ethyl 2-methyl-2-(5-nitro-1H-indol-2-yl)propanoate (4.60 g,
16.7 mmol) was dissolved in THF/water (2:1, 30 mL). LiOH.H.sub.2O
(1.40 g, 33.3 mmol) was added and the mixture was stirred at
50.degree. C. for 3 h. The mixture was made acidic by the careful
addition of 3N HCl. The product was extracted with ethylacetate and
the combined organic phases were washed with brine and dried over
magnesium sulfate to give
2-methyl-2-(5-nitro-1H-indol-2-yl)propanoic acid (4.15 g, 99%).
##STR00706##
2-Methyl-2-(5-nitro-1H-indol-2-yl)propanamide
[0554] 2-Methyl-2-(5-nitro-1H-indol-2-yl)-propanoic acid (4.12 g,
16.6 mmol) was dissolved in acetonitrile (80 mL). EDC (3.80 g,
0.020 mmol), HOBt (2.70 g, 0.020 mmol), Et.sub.3N (6.9 mL, 0.050
mmol) and ammonium chloride (1.34 g, 0.025 mmol) were added and the
mixture was stirred overnight at room temperature. Water was added
and the mixture was extracted with ethylacetate. Combined organic
phases were washed with brine, dried over magnesium sulfate and
dried to give 2-methyl-2-(5-nitro-1H-indol-2-yl)propanamide (4.3 g,
99%).
##STR00707##
2-Methyl-2-(5-nitro-1H-indol-2-yl)propan-1-amine
[0555] 2-Methyl-2-(5-nitro-1H-indol-2-yl)propanamide (200 mag, 0.81
mmol) was suspended in THF (5 ml) and cooled to 0.degree. C.
Borane-THF complex solution (1.0 M, 2.4 mL, 2.4 mmol) was added
slowly and the mixture was allowed to stir overnight at room
temperature. The mixture was cooled to 0.degree. C. and carefully
acidified with 3 N HCl. THF was evaporated off, water was added and
the mixture was washed with ethylacetate. The aqueous layer was
made alkaline with 50% NaOH and the mixture was extracted with
ethylacetate. The combined organic layers were dried over magnesium
sulfate, filtered and evaporated to give
2-methyl-2-(S-nitro-1H-indol-2-yl)propan-1-amine (82 mg, 43%).
##STR00708##
tert-Butyl 2-methyl-2-(5-nitro-1H-indol-2-yl)propylcarbamate
[0556] 2-Methyl-2-(5-nitro-1H-indol-2-yl)propan-1-amine (137 mg,
0.587 mmol) was dissolved in THF (5 mL) and cooled to 0.degree. C.
Et.sub.3N (82 .mu.L, 0.59 mmol) and di-tert-butyl dicarbonate (129
mg, 0.587 mmol) were added and the mixture was stirred at room
temperature overnight. Water was added and the mixture was
extracted with ethylacetate. The residue was purified by silica gel
chromatography (10-40% ethylacetate in hexane) to give tert-butyl
2-methyl-2-(5-nitro-1H-indol-2-yl)propylcarbamate (131 mg,
67%).
##STR00709##
tert-Butyl 2-(5-amino-1H-indol-2-yl)-2-methylpropylcarbamate
[0557] To a solution of tert-butyl
2-methyl-2-(5-nitro-1H-indol-2-yl)propylcarbamate (80 mg, 0.24
mmol) in THF (9 mL) and water (2 mL) was added ammonium formate (60
mg, 0.96 mmol) followed by 10% Pd/C (50 mg). The mixture was
stirred at room temperature for 45 minutes. Pd/C was filtered off
and the organic solvent was removed by evaporation. The remaining
aqueous phase was extracted with dichloromethane. The combined
organic phases were dried over magnesium sulfate and evaporated to
give tert-butyl 2-(5-amino-1H-indol-2-yl)-2-methylpropylcarbamate
(58 mg, 80%).
##STR00710##
tert-Butyl
2-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-2-yl)-
-2-methylpropylcarbamate
[0558] tert-Butyl 2-(5-amino-1H-indol-2-yl)-2-methylpropylcarbamate
(58 mg, 0.19 mmol),
1-(benzo[d)[1,3]dioxol-6-yl)cyclopropanecarboxylic acid (47 mg,
0.23 mmol), EDC (45 mg, 0.23 mmol), HOBt (31 mg, 0.23 mmol) and
Et.sub.3N (80 .mu.L, 0.57 mmol) were dissolved in DMF (4 mL) and
stirred overnight at room temperature. The mixture was diluted with
water and extracted with ethylacetate. The combined organic phases
were dried over magnesium sulfate and evaporated to dryness. The
residue was purified by silica gel chromatography (10-30%
ethylacetate in hexane) to give tert-butyl
2-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-2-yl)-
-2-methylpropylcarbamate (88 mg, 94%). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.32 (s; 1H), 7.62 (d, J=1.5 Hz, 1H), 7.18-7.16
(m, 2H), 7.02-6.94 (m, 3H), 6.85 (d, J=7.8 Hz, 1H), 6.19 (d, J=1.5
Hz, 1H), 6.02 (s, 2H), 4.54 (m, 1H), 3.33 (d, J=6.2 Hz, 2H), 1.68
(dd, J=3.7, 6.8 Hz, 2H), 1.36 (s, 9H), 1.35 (s, 6H), 1.09 (dd,
J=3.7, 6.8 Hz, 2H).
Example 72
(R)--N-(2-tert-Butyl-1-(2,3-dihydroxypropyl)-1H-indol-5-yl)-1-(2,2-difluor-
obenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
##STR00711##
[0559]
(R)-2-tert-Butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-5-nitro-
-1H-indole
[0560] To a stirred solution of
(S)-(2,2-dimethyl-1,3-dioxolan-4-yl)methyl 4-methylbenzenesulfonate
(1.58 g, 5.50 mmol) in anhydrous DMF (10 mL) under nitrogen gas was
added 2-tert-butyl-5-nitro-1H-indole (1.00 g, 4.58 mmol) followed
by Cs.sub.2CO.sub.3 (2.99 g, 9.16 mol). The mixture was stirred and
heated at 80.degree. C. under nitrogen gas. After 20 hours, 50%
conversion was observed by LCMS. The reaction mixture was
re-treated with Cs.sub.2CO.sub.3 (2.99 g, 9.16 mol) and
(S)-(2,2-dimethyl-1,3-dioxolan-4-yl)methyl 4-methylbenzenesulfonate
(1.58 g, 5.50 mmol) and heated at 80.degree. C. for 24 hours. The
reaction mixture was cooled to room temperature. The solids were
filtered and washed with ethyl acetate and hexane (1:1). The layers
were separated and the organic layer was washed with water
(2.times.10 mL) and brine (2.times.10 mL). The organic layer was
dried over Na.sub.2SO.sub.4, filtered and evaporated under reduced
pressure. The residue was purified by column chromatography on
silica gel (dichloromethane/hexane=1.5/1) to give
(R)-2-tert-butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-5-nitro--
1H-indole (1.0 g, 66%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
8.48 (d, J=2.2 Hz, 1H), 8.08 (dd, J=2.2, 9.1 Hz, 1H), 7.49 (d,
J=9.1 Hz, 1H), 6.00 (s, 1H), 4.52-4.45 (m, 3H), 4.12 (dd, J=6.0,
8.6 Hz, 1H), 3.78 (dd, J=6.0, 8.6 Hz, 1H), 1.53 (s, 3H), 1.51 (s,
9H), 1.33 (s, 3H).
##STR00712##
(R)-2-tert-Butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl-1H-indol-5-ami-
ne
[0561] To a stirred solution of
(R)-2-tert-butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-5-nitro-1H-in-
dole (1.0 g, 3.0 mmol) in ethanol (20 mL) and water (5 mL) was
added ammonium formate (0.76 g, 12 mmol) followed by slow addition
of 10% palladium on carbon (0.4 g). The mixture was stirred at room
temperature for 1 h. The reaction mixture was filtered through a
plug of celite and rinsed with ethyl acetate. The filtrate was
evaporated under reduced pressure and the crude product was
dissolved in ethyl acetate. The organic layer was washed with water
(2.times.5 mL) and brine (2.times.5 mL). The organic layer was
dried over Na.sub.2SO.sub.4, filtered and evaporated under reduced
pressure to give
(R)-2-tert-butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl-1H-indol-5-ami-
ne (0.89 g, 98%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.04
(d, J=4 Hz, 1H), 6.70 (d, J=2.2 Hz, 1H), 6.48 (dd, J=2.2, 8.6 Hz,
1H), 6.05 (s, 1H), 4.38-4.1 (m, 2H), 4.21 (dd, J=7.5, 16.5 Hz, 1H),
3.87 (dd, J=6.0, 8.6 Hz, 1H), 3.66 (dd, J=6.0, 8.6 Hz, 1H), 3.33
(br s, 2H), 1.40 (s, 3H), 1.34 (s, 9H), 1.25 (s, 3H).
##STR00713##
N--((R)-2-tert-Butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-indol--
5-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0562] To 1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid
(0.73 g, 3.0 mmol) was added thionyl chloride (660 .mu.L, 9.0 mmol)
and DMF (20 .mu.L) at room temperature. The mixture was stirred for
30 minutes before the excess thionyl chloride was evaporated under
reduced pressure. To the resulting acid chloride, dichloromethane
(6.0 mL) and Et.sub.3N (2.1 mL, 15 mmol) were added. A solution of
(R)-2-tert-butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl-1H-indol-5-ami-
ne (3.0 mmol) in dichloromethane (3.0 mL) was added to the cooled
acid chloride solution. After addition, the reaction mixture was
stirred at room temperature for 45 minutes. The reaction mixture
was filtered and the filtrate was evaporated under reduced
pressure. The residue was purified by column chromatography on
silica gel (ethyl acetate/hexane=3/7) to give
N--((R)-2-tert-butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-indol--
5-yl)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
(1.33 g, 84%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.48 (d,
J=2 Hz, 1H), 7.31 (dd, J=2, 8 Hz, 1H), 7.27 (dd, J=2, 8 Hz, 1H),
7.23 (d, J=8 Hz, 1H), 7.14 (d, J=8 Hz, 1H), 7.02 (dd, J=2, 8 Hz,
1H), 6.92 (br s, 1H), 6.22 (s, 1H), 4.38-4.05 (m, 3H), 3.91 (dd,
J=5, 8 Hz, 1H), 3.75 (dd, J=5, 8 Hz, 1H), 2.33 (q, J=8 Hz, 2H),
1.42 (s, 3H), 1.37 (s, 9H), 1.22 (s, 3H), 1.10 (q, J=8 Hz, 2H).
##STR00714##
N--((R)-2-tert-Butyl-1-((2,3-dihydroxypropyl)-1H-indol-5-yl)-1-(2,2-diflu-
orobenzo-[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0563] To a stirred solution of
N-(2-tert-butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-indol-5-yl)-
-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
(1.28 g, 2.43 mmol) in methanol (34 mL) and water (3.7 mL) was
added para-toluenesulfonic acid-hydrate (1.87 g, 9.83 mmol). The
reaction mixture was stirred and heated at 80.degree. C. for 25
minutes. The solvent was evaporated under reduced pressure. The
crude product was dissolved in ethyl acetate. The organic layer was
washed with saturated aqueous NaHCO.sub.3 (2.times.10 mL) and brine
(2.times.10 mL). The organic layer was dried over Na.sub.2SO.sub.4,
filtered and evaporated under reduced pressure. The residue was
purified by column chromatography on silica gel (ethyl
acetate/hexane=13/7) to give
N--((R)-2-tert-butyl-1-((2,3-dihydroxypropyl)-1H-indol-5-yl)-1-(2,2-diflu-
orobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide (0.96 g, 81%).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.50 (d, J=2 Hz, 1H),
7.31 (dd, J=2, 8 Hz, 1H), 7.27 (dd, J=2, 8 Hz, 1H), 7.23 (d, J=8
Hz, 1H), 7.14 (d, J=8 Hz, 1H), 7.02 (br s, 1H), 6.96 (dd, J=2, 8
Hz, 1H), 6.23 (s, 1H), 4.35 (dd, J=8, 15 Hz, 1H), 4.26 (dd, J=4, 15
Hz, 1H), 4.02-3.95 (m, 1H), 3.60 (dd, J=4, 11 Hz, 1H), 3.50 (dd,
J=5, 11 Hz, 1H), 1.75 (q, J=8 Hz, 3H), 1.43 (s, 9H), 1.14 (q, J=8
Hz, 3H).
Example 73
3-(2-tert-Butyl-S-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecar-
boxamido)-1H-indol-1-yl)-2-hydroxypropanoic acid
##STR00715##
[0564]
3-(2-tert-Butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxo-5-yl)cycloprop-
anecarbox-amido)-1H-indol-1-yl)-2-oxopropanoic acid
[0565] To a solution of
N-(2-tert-butyl-1-(2,3-dihydroxypropyl)-1H-indol-5-yl)-1-(2,2-difluoroben-
zo[d][1,3]dioxol-5-yl)cyclopropane-carboxamide (97 mg, 0.20 mmol)
in DMSO (1 mL) was added Dess-Martin periodinane (130 mg, 0.30
mmol). The mixture was stirred at room temperature for 3 h. The
solid was filtered off and washed with EtOAc. The filtrate was
partitioned between EtOAc and water. The aqueous layer was
extracted with EtOAc twice and the combined organic layers were
washed with brine and dried over MgSO.sub.4. After the removal of
solvent, the residue was purified by preparative TLC to yield.
3-(2-tert-butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropaneca-
rboxamido)-1H-indol-1-yl)-2-oxopropanoic acid that was used without
further purification.
##STR00716##
3-(2-tert-Butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropaneca-
rbox-amido)-1H-indol-1-yl)-2-hydroxypropanoic acid
[0566] To a solution of
3-(2-tert-butyl-S-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropaneca-
rboxamido)-1H-indol-1-yl)-2-oxopropanoic acid (50 mg, 0.10 mmol) in
MeOH (1 mL) was added NaBH.sub.4 (19 mg, 0.50 mmol) at 0.degree. C.
The mixture was stirred at room temperature for 15 min. The
resulting mixture was partitioned between EtOAc and water. The
aqueous layer was extracted with EtOAc twice and the combined
organic layers were washed with brine and dried over anhydrous
MgSO.sub.4. After the removal of the solvent, the residue was taken
up in DMSO and purified by preparative LC/MS to give
3-(2-tert-butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cycloprop-
anecarboxamido)-1H-indol-1-yl)-2-hydroxypropanoic acid. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.36 (s), 7.27-7.23 (m, 2H),
7.15-7.11 (m, 2H), 6.94 (d, J=8.5 Hz, 1H), 6.23 (s, 1H), 4.71 (s,
3H), 4.59 (q, J=10.3 Hz, 1H), 4.40-4.33 (m, 21H), 1.70 (d, J=1.9
Hz, 2H), 1.15 (q, J=4.0 Hz, 2H). .sup.13C NMR (400 MHz, CDCl.sub.3)
.delta. 173.6, 173.1, 150.7, 144.1, 143.6, 136.2, 135.4, 134.3,
131.7, 129.2, 129.0, 127.6, 126.7, 116.6, 114.2, 112.4, 110.4,
110.1, 99.7, 70.3, 48.5, 32.6, 30.9, 30.7, 16.8. MS (ESI) m/e
(M+H.sup.+) 501.2.
Example 74
(R)--N-(2-tert-Butyl-1-(2,3-dihydroxypropyl)-1H-indol-5-yl)-1-(2,2-dideute-
riumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
##STR00717##
[0567] Methyl 1-(3,4-dihydroxyphenyl)cyclopropanecarboxylate
[0568] To a solution of
1-(3,4-dihydroxyphenyl)cyclopropanecarboxylic acid (190 mg, 1.0
mmol) in MeOH (3 mL) was added 4-methylbenzenesulfonic acid (19 mg,
0.10 mmol). The mixture was heated at 80.degree. C. overnight. The
reaction mixture was concentrated in vacuo and partitioned between
EtOAc and water. The aqueous layer was extracted with EtOAc twice
and the combined organic layers were washed with sat. NaHCO.sub.3
and brine and dried over MgSO.sub.4. After the removal of solvent,
the residue was dried in vacuo to yield methyl
1-(3,4-dihydroxyphenyl)cyclopropanecarboxylate (190 mg, 91%) that
was used without further purification. .sup.1H NMR (400 MHz,
DMSO-d.sup.6) .delta. 6.76-6.71 (m, 2H), 6.66 (d, J=7.9 Hz, 1H),
3.56 (s, 3H), 1.50 (q, J=3.6 Hz, 2H), 1.08 (q, J=3.6 Hz, 2H).
##STR00718##
Methyl
1-(2,2-dideuteriumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylate
[0569] To a solution of methyl
1-(3,4-dihydroxyphenyl)cyclopropanecarboxylate (21 mg, 0.10 mmol)
and CD.sub.2Br.sub.2 (35 mg, 0.20 mmol) in DMF (0.5 mL) was added
Cs.sub.2CO.sub.3 (19 mg, 0.10 mmol). The mixture was heated at
120.degree. C. for 30 min. The reaction mixture was partitioned
between EtOAc and water. The aqueous layer was extracted with EtOAc
twice and the combined organic layers were washed with 1N NaOH and
brine before being dried over MgSO.sub.4. After the removal of
solvent, the residue was dried in vacuo to yield methyl
1-(2,2-dideuteriumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylate
(22 mg) that was used without further purification. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 6.76-6.71 (m, 2H), 6.66 (d, J=7.9 Hz,
1H), 3.56 (s, 3H), 1.50 (q, J=3.6 Hz, 2H), 1.08 (q, J=3.6 Hz,
2H).
##STR00719##
1-(2,2-Dideuteriumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic
acid
[0570] To a solution of methyl
1-(2,2-dideuteriumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylate
(22 mg, 0.10 mmol) in THF (0.5 mL) was added NaOH (1N, 0.25 mL,
0.25 mmol). The mixture was heated at 80.degree. C. for 2 h. The
reaction mixture was partitioned between EtOAc and 1N NaOH. The
aqueous layer was extracted with EtOAc twice, neutralized with 1N
HCl and extracted with EtOAc twice. The combined organic layers
were washed with brine and dried over MgSO.sub.4. After the removal
of solvent, the residue was dried in vacuo to yield
1-(2,2-dideuteriumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxyli- c
acid (21 mg) that was used without further purification.
##STR00720##
(R)--N-(2-tert-Butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-indol--
5-yl)-1-(2,2-dideuteriumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0571] To a solution of
1-(2,2-dideuteriumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic
acid (21 mg, 0.10 mmol),
(R)-2-tert-butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-indol-5-am-
ine (30 mg, 0.10 mmol), HATU (42 mg, 0.11 mol) in DMF (1 mL) was
added triethylamine (0.030 mL, 0.22 mmol). The mixture was heated
at room temperature for 5 min. The reaction mixture was partitioned
between EtOAc and water. The aqueous layer was extracted with EtOAc
twice and the combined organic layers were washed with 1N NaOH, 1N
HCl, and brine before being dried over MgSO.sub.4. After the
removal of solvent, the residue was purified by column
chromatography (20-40% ethyl acetate/hexane) to yield
(R)--N-(2-tert-butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-indol--
5-yl)-1-(2,2-dideuteriumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
(24 mg, 49% from methyl
1-(3,4-dihydroxyphenyl)cyclopropanecarboxylate). MS (ESI) m/e
(M+H.sup.+) 493.5.
##STR00721##
(R)--N-(2-tert-Butyl-1-(2,3-dihydroxypropyl)-1H-Indol-5-yl)-1-(2,2-dideut-
erium-benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0572] To a solution of
(R)--N-(2-tert-butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-indol--
5-yl)-1-(2,2-dideuterium-benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
(24 mg, 0.050 mmol), in methanol (0.5 mL) and water (0.05 mL) was
added 4-methylbenzenesulfonic acid (2.0 mg, 0.010 mmol). The
mixture was heated at 80.degree. C. for 30 min. The reaction
mixture was partitioned between EtOAc and water. The aqueous layer
was extracted with EtOAc twice and the combined organic layers were
washed with sat. NaHCO.sub.3 and brine before being dried over
MgSO.sub.4. After the removal of solvent, the residue was purified
by preparative HPLC to yield
(R)--N-(2-tert-butyl-1-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-1H-indol--
5-yl)-1-(2,2-dideuteriumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
(12 mg, 52%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.44 (d,
J=2.0 Hz, 1H), 7.14 (dd, J=22.8, 14.0 Hz, 211H), 6.95-6.89 (m, 2H),
6.78 (d, J=7.8 Hz, 1H), 6.14 (s, 1H), 4.28 (dd, J=15.1, 8.3 Hz,
1H), 4.19 (dd, J=15.1, 4.5 Hz, 1H), 4.05 (q, J=7.1 Hz, 1H), 3.55
(dd, J=11.3, 4.0 Hz, 1H), 3.45 (dd, J=11.3, 5.4 Hz, 1H), 1.60 (q,
J=3.5 Hz, 2H), 1.35 (s, 9H), 1.02 (q, J=3.5 Hz, 2H). .sup.13C NMR
(400 MHz, CDCl.sub.3) .delta. 171.4, 149.3, 147.1, 146.5. 134.8,
132.3, 129.2, 126.5, 123.6, 114.3, 111.4, 110.4, 109.0, 107.8,
98.5, 70.4, 63.1, 46.6, 31.6, 30.0, 29.8, 15.3. MS (ESI) m/e
(M+H.sup.+) 453.5.
[0573] It is further noted that the mono-deuterated analogue for
this compound can be synthesized by substitution the reagent
CHDBR.sub.2 for CD.sub.2BR.sub.2 and following the procedures
described in example 74. Furthermore, deuterated analogues of the
compounds as described herein such as of formula I can be produced
using known synthesitc methods as well as the methodology described
herein. The deuterated analogues include both di and
mono-deuterated analogues of the compounds of the present
invention. The di and mono deuterated analogues of the compounds
exhibit measurable activity when tested using the assays described
below.
Example 75
4-(5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-2-yl)--
4-methylpentanoic acid
##STR00722##
[0574]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-(4-cyano-2-methylbutan-2-yl)-1H-i-
ndol-5-yl)cyclopropanecarboxamide
[0575] To 1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid
(0.068 g, 0.33 mmol) was added thionyl chloride (72 .mu.L, 0.99
mmol) and DMF (20 .mu.L) at room temperature. The mixture was
stirred for 30 minutes before the excess thionyl chloride was
evaporated under reduced pressure. To the resulting acid Chloride,
dichloromethane (0.5 mL) and Et.sub.3N (230 .mu.L, 1.7 mmol) were
added. A solution of
4-(5-amino-1H-indol-2-yl)-4-methylpentanenitrile (0.33 mmol) in
dichloromethane (0.5 mL) was added to the acid chloride solution
and the mixture was stirred at room temperature for 1.5 h. The
resulting mixture was diluted with dichloromethane and washed with
1 N HCl (2.times.2 mL), saturated aqueous NaHCO.sub.3 (2.times.2
mL) and brine (2.times.2 mL). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4 and evaporated under reduced pressure to
give
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-(4-cyano-2-methylbutan-2-yl)-1H-indol-5-
-yl)cyclopropanecarboxamide.
##STR00723##
4-(5-(1-(Benzo[d]1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-2-yl)--
4-methylpentanoic acid
[0576] A mixture of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-(4-cyano-2-methylbutan-2-yl)-1H-indol-5-
-yl)cyclopropanecarboxamide (0.060 g, 0.15 mmol) and KOH (0.081 g,
1.5 mmol) in 50% EtOH/water (2 mL) was heated in the microwave at
100.degree. C. for 1 h. The solvent was evaporated under reduced
pressure. The crude product was dissolved in DMSO (1 mL), filtered,
and purified by reverse phase preparative HPLC to give
4-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-2-yl)-
-4-methylpentanoic acid. .sup.1H NMR (400 MHz, DMSO-d6) .delta.
11.98 (s, 1H), 10.79 (s, 1H), 8.44 (s, 1H), 7.56 (s, 1H), 7.15 (d,
J=8.6 Hz, 1H), 7.03-6.90 (m, 4H), 6.05 (s, 1H), 6.02 (s, 2H),
1.97-1.87 (m, 4H), 1.41-1.38 (m, 2H), 1.30 (s, 6H), 1.04-1.02 (m,
2H).
Example 76
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-(1-hydroxypropan-2-yl)-1H-indol-5-yl)cyc-
lopropanecarboxamide
##STR00724##
[0577] 2-(5-Nitro-1H-indol-2-yl)propan-1-ol
[0578] To a cooled solution of LiAlH.sub.4 (1.0 M in THF, 1.2 mL,
1.2 mmol) in THF (5.3 mL) at 0.degree. C. was added a solution of
ethyl 2-(5-nitro-1H-indol-2-yl)propanoate (0.20 g, 0.76 mmol) in
THF (3.66 mL) dropwise. After addition, the mixture was allowed to
warm up to room temperature and was stirred at room temperature for
3 h. The mixture was cooled to 0.degree. C. Water (2 mL) was slowly
added followed by careful addition of 15% NaOH (2 mL) and water (4
mL). The mixture was stirred at room temperature for 0.5 h and was
then filtered through a short plug of celite using ethyl acetate.
The organic layer was separated from the aqueous layer, dried over
Na.sub.2SO.sub.4, filtered and evaporated under reduced pressure.
The residue was purified by column chromatography on silica gel
(ethyl acetate/hexane=1/1) to give
2-(5-nitro-1H-indol-2-yl)propan-1-ol (0.14 g, 81%).
##STR00725##
2-(5-Amino-1H-indol-2-yl)propan-1-ol
[0579] To a solution of 2-(5-nitro-1H-indol-2-yl)propan-1-ol (0.13
g, 0.60 mmol) in ethanol (5 mL) was added tin chloride dihydrate
(0.67 g, 3.0 mmol). The mixture was heated in the microwave at
120.degree. C. for 1 h. The mixture was diluted with ethyl acetate
before water and saturated aqueous NaHCO.sub.3 were added. The
reaction mixture was filtered through a plug of celite using ethyl
acetate. The organic layer was separated from the aqueous layer,
dried over Na.sub.2SO.sub.4, filtered and evaporated under reduced
pressure to give 2-(5-amino-1H-indol-2-yl)propan-1-ol (0.093 g,
82%).
##STR00726##
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-(1-hydroxypropan-2-yl)-1H-indol-5-yl)cy-
clopropanecarboxamide
[0580] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid (0.10 g,
0.49 mmol) in acetonitrile (2.0 mL) were added HBTU (0.185 g, 0.49
mmol) and Et.sub.3N (205 .mu.L, 1.47 mmol) at room temperature. The
mixture was allowed to stir at room temperature for 10 minutes
before a slurry of 2-(5-amino-1H-indol-2-yl)propan-1-ol (0.093 g,
0.49 mmol) in acetonitrile (2.7 mL) was added. After addition, the
reaction mixture was stirred at room temperature for 5.5 h. The
solvent was evaporated under reduced pressure and the residue was
dissolved in dichloromethane. The organic layer was washed with 1 N
HCl (1.times.3 mL) and saturated aqueous NaHCO.sub.3 (1.times.3
mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered
and evaporated under reduced pressure. The crude material was
purified by column chromatography on silica gel (ethyl
acetate/hexane=13/7) to give
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-(1-hydroxypropan-2-yl)-1H-indol-5-yl)cy-
clopropanecarboxamide (0.095 g, 51%). .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 10.74 (s, 1H), 8.38 (s, 1H), 7.55 (s, 1H), 7.14
(d, J=8.6 Hz, 1H), 7.02-6.90 (m, 4H), 6.06 (s, 1H), 6.02 (s, 2H),
4.76 (t, J=5.3 Hz, 1H), 3.68-3.63 (m, 1H), 3.50-3.44 (m, 1H),
2.99-2.90 (m, 1H), 1.41-1.38 (m, 2H), 1.26 (d, J=7.0 Hz, 3H),
1.05-1.02 (m, 2H).
Example 77
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)-N-methylcyclop-
ropanecarboxamide
##STR00727##
[0581]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)-N-methy-
lcyclopropanecarboxamide
[0582] 2-tert-Butyl-N-methyl-1H-indol-5-amine (20.2 mg, 0.100 mmol)
and 1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic acid (20.6
mg, 0.100 mmol) were dissolved in N,N-dimethylformamide (1 mL)
containing triethylamine (42.1 .mu.L, 0.300 mmol) and a magnetic
stir bar. O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (42 mg, 0.11 mmol) was added to the mixture and
the resulting solution was allowed to stir for 16 h at 80.degree.
C. The crude product was then purified by preparative HPLC
utilizing a gradient of 0-99% acetonitrile in water containing
0.05% trifluoroacetic acid to yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)-N-methy-
lcyclopropanecarboxamide. ESI-MS m/z calc. 390.2, found 391.3
(M+1).sup.+. Retention time of 3.41 minutes.
Example 78
N-(2-tert-Butyl-1-methyl-1H-indol-5-yl)-1-(benzo[d][1,3]dioxol-6-yl)-N-met-
hylcyclopropanecarboxamide
##STR00728##
[0584] Sodium hydride (0.028 g, 0.70 mmol, 60% by weight dispersion
in oil) was slowly added to a stirred solution of
N-(2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]dioxol-6-yl)cyclopropaneca-
rboxamide (0.250 g, 0.664 mmol) in a mixture of 4.5 mL of anhydrous
tetrahydrofuran (THF) and 0.5 mL of anhydrous N,N-dimethylformamide
(DMF). The resulting suspension was allowed to stir for 2 minutes
and then iodomethane (0.062 mL, 1.0 mmol) was added to the reaction
mixture. Two additional aliquots of sodium hydride and iodomethane
were required to consume all of the starting material which was
monitored by LC/MS. The crude reaction product was evaporated to
dryness, redissolved in a minimum of DMF and purified by
preparative LC/MS chromatography to yield the pure product (0.0343
g, 13%) ESI-MS m/z calc. 404.2, found 405.3 (M+1).sup.+. Retention
time of 3.65 minutes.
Example 79
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-(hydroxymethyl)-1H-indol-5-yl)cyclopropa-
necarboxamide
##STR00729##
[0586] Ethyl
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indole-2-carbo-
xylate (1.18 g, 3.0 mmol) was added to a solution of LiBH.sub.4
(132 mg, 6.0 mmol) in THF (10 mL) and water (0.1 mL). The mixture
was allowed to stir for 16 h at 25.degree. C. before it was
quenched with water (10 mL) and slowly made acidic by addition of 1
N HCl. The mixture was extracted with three 50-mL portions of ethyl
acetate. The organic extracts were dried over Na.sub.2SO.sub.4 and
evaporated to yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-(hydroxymethyl)-1H-indol-5-yl)cycloprop-
anecarboxamide (770 mg, 73%). A small amount was further purified
by reverse phase HPLC. ESI-MS m/z calc. 350.4, found 351.3
(M+1).sup.+; retention time 2.59 minutes.
Example 80
5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-N-tert-butyl-1H-in-
dole-2-carboxamide
##STR00730##
[0587]
5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indole-2-
-carboxylic acid
[0588] Ethyl
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indole-2-carbo-
xylate (392 mg, 1.0 mmol) and LiOH (126 mg, 3 mmol) were dissolved
in H.sub.2O (5 mL) and 1,4-dioxane (3 mL). The mixture was heated
in an oil bath at 100.degree. C. for 24 hours before it was cooled
to room temperature. The mixture was acidified with 1N HCl and it
was extracted with three 20 mL portions of dichloromethane. The
organic extracts were dried over Na.sub.2SO.sub.4 and evaporated to
yield
5-(1-(benzo[d][1,3]-dioxol-5-yl)cyclopropanecarboxamido)-1H-indole-2-carb-
oxylic acid (302 mg, 83%). A small amount was further purified by
reverse phase HPLC. ESI-MS m/z calc. 364.1, found 365.1 (M+1)*;
retention time 2.70 minutes.
##STR00731##
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-N-tert-butyl-1H-i-
ndole-2-carboxamide
[0589]
5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropane-carboxamido)-1H-indole--
2-carboxylic acid (36 mg, 0.10 mmol) and 2-methylpropan-2-amine
(8.8 mg, 0.12 mmol) were dissolved in N,N-dimethylformamide (1.0
mL) containing triethylamine (28 .mu.L, 0.20 mmol).
O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (46 mg, 0.12 mmol) was added to the mixture and
the resulting solution was allowed to stir for 3 hours. The mixture
was filtered and purified by reverse phase HPLC to yield
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-N-tert-butyl-1H-i-
ndole-2-carboxamide. ESI-MS m/z calc. 419.2, found 420.3
(M+1).sup.+; retention time 3.12 minutes.
Example 81
N-(3-Amino-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclopr-
opanecarboxamide
##STR00732##
[0591] A solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)cyclopropane
carboxamide (50 mg, 0.13 mmol) was dissolved in AcOH (2 mL) and
warmed to 45.degree. C. To the mixture was added a solution of
NaNO.sub.2 (9 mg) in H.sub.2O (0.03 mL). The mixture was allowed to
stir for 30 min at 45.degree. C. before the precipitate was
collected and washed with Et.sub.2O. This material was used in the
next step without further purification. To the crude material,
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-nitroso-1H-indol-5-yl)cycl-
opropanecarboxamide, was added AcOH (2 mL) and Zn dust (5 mg). The
mixture was allowed to stir for 1 h at ambient temperature. EtOAc
and H.sub.2O were added to the mixture. The layers were separated
and the organic layer was washed with sat. aq. NaHCO.sub.3, dried
over MgSO.sub.4, and concentrated in vacuo. The residue was taken
up in DMF (1 mL) and was purified using prep-HPLC. LCMS: m/z 392.3;
retention time of 2.18 min.
Example 82
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-(methylsulfonyl)-1H-indol-5-
-yl)cyclopropanecarboxamide
##STR00733##
[0592]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-(methylsulfonyl)-1H--
indol-5-yl)cyclopropanecarboxamide
[0593] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)cyclopropaneca-
rboxamide (120 mg, 0.31 mmol) in anhydrous DMF-THF (3.3 mL, 1:9)
was added NaH (60% in mineral oil, 49 mg, 1.2 mmol) at room
temperature. After 30 min under N.sub.2, the suspension was cooled
down to -15.degree. C. and a solution of methanesulfonyl chloride
(1.1 eq.) in DMF (0.5 mL) was added dropwise. The reaction mixture
was stirred for 30 min at -15.degree. C. then for 6 h at room
temperature. Water (0.5 mL) was added at 0.degree. C., solvent was
removed, and the residue was diluted with MeOH, filtrated and
purified by preparative HPLC to give
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-(methylsulfonyl)-1H-indol--
5-yl)cyclopropanecarboxamide. .sup.1H NMR (400 MHz, DMSO) .delta.
11.6 (s, 1H), 8.7 (s, 1H), 7.94 (d, J=1.7 Hz, 1H), 7.38 (d, J=8.7
Hz, 1H), 7.33 (dd, J1=1.9 Hz, J2=8.7 Hz, 1H), 7.03 (d, J=1.7 Hz,
1H), 6.95 (dd, J1=1.7 Hz, J2=8.0 Hz, 1H), 6.90 (d, J=8.0 Hz, 1H),
6.02 (s, 2H), 3.07 (s, 3H), 1.56-1.40 (m, 9H), 1.41 (dd, J=4.0 Hz,
J2=6.7 Hz, 2H), 1.03 (dd, J1=4.0 Hz, J2=6.7 Hz, 2H). MS (ESI) m/e
(M+H.sup.+) 455.5.
Example 83
1-(Benzo[d][1,3]dioxol-5-yl)-N-(3-phenyl-1H-indol-5-yl)cyclopropane
carboxamide
##STR00734##
[0594]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(3-bromo-1H-indol-5-yl)cyclopropanec-
arboxamide
[0595] Freshly recrystallized N-bromosuccinimide (0.278 g, 1.56
mmol) was added portionwise to a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(1H-indol-5-yl)cyclopropanecarboxamide
(0.500 g, 1.56 mmol) in N,N-dimethylformamide (2 mL) over 2
minutes. The reaction mixture was protected from light and was
stirred bar for 5 minutes. The resulting green solution was poured
into 40 mL of water. The grey precipitate which formed was filtered
and washed with water to yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(3-bromo-1H-indol-5-yl)cyclopropanecarboxa-
mide (0.564 g, 91%). ESI-MS m/z calc. 398.0, found 399.3
(M+1).sup.+. Retention time of 3.38 minutes. .sup.1H NMR (400 MHz,
DMSO-d6) 11.37 (s, 1H), 8.71 (s, 1H), 7.67 (d, J=1.8 Hz, 1H), 7.50
(d, J=2.6 Hz, 1H), 7.29 (d, J=8.8 Hz, 1H), 7.22 (dd, J=2.0, 8.8 Hz,
1H), 7.02 (d, J=1.6 Hz, 1H), 6.96-6.88 (m, 2H), 6.03 (s, 2H),
1.43-1.40 (m, 2H), 1.09-1.04 (m, 2H).
##STR00735##
1-(Benzo[d][1,3]dioxol-5-yl)-N-(3-phenyl-1H-indol-5-yl)cyclopropanecarbox-
amide
[0596] Phenyl boronic acid (24.6 mg, 0.204 mmol) was added to a
solution of
1-(benzo[d][1,3]-dioxol-5-yl)-N-(3-bromo-1H-indol-5-yl)cyclopropanecar-
boxamide (39.9 mg, 0.100 mmol) in ethanol (1 mL) containing
FibreCat 1001 (6 mg) and 1M aqueous potassium carbonate (0.260 mL).
The reaction mixture was then heated at 130.degree. C. in a
microwave reactor for 20 minutes. The crude product was then
purified by preparative HPLC utilizing a gradient of 0-99%
acetonitrile in water containing 0.05% trifluoroacetic acid to
yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(3-phenyl-1H-indol-5-yl)cyclopropane
carboxamide. ESI-MS m/z calc. 396.2, found 397.3 (M+1).sup.+.
Retention time of 3.52 minutes. .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 11.27 (d, J=1.9 Hz, 1H), 8.66 (s, 1H), 8.08 (d, J=1.6 Hz,
1H), 7.65-7.61 (m, 3H), 7.46-7.40 (m, 2H), 7.31 (d, J=8.7 Hz, 1H),
7.25-7.17 (m, 2H), 7.03 (d, J=1.6 Hz, 1H), 6.98-6.87 (m, 2H), 6.02
(s, 2H), 1.43-1.39 (m, 2H), 1.06-1.02 (m, 2H).
Example 84
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-cyano-1H-indol-5-yl)cyclopr-
opanecarboxamide
##STR00736##
[0597]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-formyl-1H-indol-5-yl-
)cyclopropane-carboxamide
[0598] POCl.sub.3 (12 g, 80 mmol) was added dropwise to DMF (40 mL)
held at -20.degree. C. After the addition was complete, the
reaction mixture was allowed to warm to 0.degree. C. and was
stirred for 1 h.
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)cyclopropaneca-
rboxamide (3.0 g, 8.0 mmol) was added and the mixture was warmed to
25.degree. C. After stirring for 30 minutes the reaction mixture
was poured over ice and stirred for 2 h. The mixture was then
heated at 100.degree. C. for 30 min. The mixture was cooled and the
solid precipitate was collected and washed with water. The solid
was then dissolved in 200 mL dichloromethane and washed with 200 mL
of a saturated aq. NaHCO.sub.3. The organics were dried over
Na.sub.2SO.sub.4 and evaporated to yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-formyl-1H-indol-5-yl)cyclo-
propane-carboxamide (2.0 g, 61%). ESI-MS m/z calc. 404.5, found
405.5 (M+1).sup.+; retention time 3.30 minutes. .sup.1H NMR (400
MHz, DMSO-d6) .delta. 11.48 (s, 1H), 10.39 (s, 1H), 8.72 (s, 1H),
8.21 (s, 1H), 7.35-7.31 (m, 2H), 7.04-7.03 (m, 1H), 6.97-6.90 (m,
2H), 6.03 (s, 2H), 1.53 (s, 9H), 1.42-1.39 (m, 2H), 1.05-1.03 (m,
2H).
##STR00737##
(Z)-1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-((hydroxyimino)methy)--
1H-indol-5-yl)cyclopropanecarboxamide
[0599] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-formyl-1H-indol-5-yl)cyclo-
propanecarboxamide (100 mg, 0.25 mmol) in dichloromethane (5 mL)
was added hydroxylamine hydrochloride (21 mg, 0.30 mmol). After
stirring for 48 h, the mixture was evaporated to dryness and
purified by column chromatography (0-100% ethyl acetate/hexanes) to
yield
(Z)-1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-((hydroxyimino)methyl)-
-1H-indol-5-yl)cyclopropanecarboxamide (81 mg, 77%). ESI-MS m/t
calc. 419.5, found 420.5 (M+1).sup.+; retention time 3.42 minutes.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.86 (s, 0.5H), 10.55 (s,
0.5H), 8.56-8.50 (m, 2H), 8.02 (m, 1H), 7.24-7.22 (m, 1H),
7.12-7.10 (in, 1H), 7.03 (m, 11H), 6.96-6.90 (m, 2H), 6.03 (s, 2H),
1.43 (s, 9H), 1.40-1.38 (m, 2H), 1.04-1.01 (m, 2H).
##STR00738##
1-Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-cyano-1H-indol-5-yl)cyclopr-
opane-carboxamide
[0600]
(Z)-1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-((hydroxyimino)--
methyl)-1H-indol-5-yl)cyclopropanecarboxamide (39 mg, 0.090 mmol)
was dissolved in acetic anhydride (1 mL) and heated at reflux for 3
h. The mixture was cooled in an ice bath and the precipitate was
collected and washed with water. The solid was further dried under
high vacuum to yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-cyano-1H-indol-5-yl)cyclop-
ropanecarboxamide. ESI-MS m/z calc. 401.5, found 402.5 (M+1).sup.+;
retention time 3.70 minutes. .sup.1H NMR (400 MHz, DMSO-d6) .delta.
11.72 (s, 1H), 8.79 (s, 1H), 7.79 (s, 1H), 7.32 (m, 2H), 7.03-7.02
(m, 1H), 6.95-6.89 (m, 2H), 6.03 (s, 2H), 1.47 (s, 9H), 1.43-1.41
(m, 2H), 1.06-1.04 (m, 2H).
Example 85
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-methyl-1H-indol-5-yl)cyclop-
ropanecarboxamide
##STR00739##
[0602] A solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)cyclopropaneca-
rboxamide (75 mg, 0.20 mmol) and iodomethane (125 .mu.L, 2.0 mmol)
in N,N-dimethylformamide (1 mL) was heated at 120.degree. C. in a
sealed tube for 24 h. The reaction was filtered and purified by
reverse phase HPLC. ESI-MS m/z calc. 390.5, found 391.3
(M+1).sup.+; retention time 2.04 minutes. .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 10.30 (s, 1H), 8.39 (s, 1H), 7.51 (m, 1H),
7.13-7.11 (m, 1H), 7.03-6.90 (m, 4H), 6.03 (s, 2H), 2.25 (s, 3H),
1.40-1.38 (m, 11H), 1.03-1.01 (m, 2H).
Example 86
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-(2-hydroxyethyl)-1H-indol-5-
-yl)cyclopropanecarboxamide
##STR00740##
[0604] Approximately 100 .mu.L of ethylene dioxide was condensed in
a reaction tube at -78.degree. C. A solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)cyclopropaneca-
rboxamide (200 mg, 0.50 mmol) and indium trichloride (20 mg, 0.10
mmol) in dichloromethane (2 mL) was added and the reaction mixture
was irradiated in the microwave for 20 min at 100.degree. C. The
volatiles were removed and the residue was purified by column
chromatography (0-100% ethyl acetate/hexanes) to give
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-(2-hydroxyethyl)-1H-indol--
5-yl)cyclopropanecarboxamide (5 mg, 3%). ESI-MS m/z calc. 420.5,
found 421.3 (M+1).sup.+; retention time 1.67 minutes. .sup.1H NMR
(400 MHz, CD.sub.3CN) .delta. 8.78 (s, 1H), 7.40 (m, 1H), 7.33 (s,
1H), 7.08 (m, 1H), 6.95-6.87 (m, 3H), 6.79 (m, 1H), 5.91 (s, 2H),
3.51 (dd, J=5.9, 7.8 Hz, 2H), 2.92-2.88 (m, 2H), 2.64 (t, J=5.8 Hz,
1H), 1.50 (m, 2H), 1.41 (s, 9H), 1.06 (m, 2H).
Example 87
2-(5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-Indol-2-yl)a-
cetic acid
##STR00741##
[0606] To a solution of ethyl
2-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-2-yl)-
acetate (0.010 g, 0.025 mmol) in THF (0.3 mL) were added
LiOH.H.sub.2O (0.002 g, 0.05 mmol) and water (0.15 mL) were added.
The mixture was stirred at room temperature for 2 h.
dichloromethane (3 mL) was added to the reaction mixture and the
organic layer was washed with 1 N HCl (2.times.1.5 mL) and water
(2.times.1.5 mL). The organic layer was dried over Na.sub.2SO.sub.4
and filtered. The filtrate was evaporated under reduced pressure to
give
2-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-2-yl)-
-acetic acid. .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.53 (s, 1H),
10.90 (s, 1H), 8.42 (s, 1H), 7.57 (s, 1H), 7.17 (d, J=8.6 Hz, 1H),
7.05-6.90 (m, 4H), 6.17 (s, 1H), 6.02 (s, 2H), 3.69 (s, 2H),
1.41-1.39 (m, 2H), 1.04-1.02 (m, 2H).
Example 88
5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butyl-1H-in-
dole-7-carboxylic acid
##STR00742##
[0608] Methyl
5-(1-(benzo[d[1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butyl-1H-in-
dole-7-carboxylate (30 mg, 0.069 mmol) was dissolved in a mixture
of 1,4-dioxane (1.5 mL) and water (2 mL) containing a magnetic star
bar and lithium hydroxide (30 mg, 0.71 mmol). The resulting
solution was stirred at 70.degree. C. for 45 minutes. The crude
product was then acidified with 2.6 M hydrochloric acid and
extracted three times with an equivalent volume of dichloromethane.
The dichloromethane extracts were combined, dried over sodium
sulfate, filtered, and evaporated to dryness. The residue was
dissolved in a minimum of N,N-dimethylformamide and then purified
by preparative HPLC using a gradient of 0-99% acetonitrile in water
containing 0.05% trifluoroacetic acid to yield
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butyl-1H-i-
ndole-7-carboxylic acid. ESI-MS m/z calc. 434.2, found 435.5.
Retention time of 1.85 minutes. .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 13.05 (s, 1H), 9.96 (d, J=1.6 Hz, 1H), 7.89 (d, J=1.9 Hz,
1H), 7.74 (d, J=2.0 Hz, 1H), 7.02 (d, J=1.6 Hz, 1H), 6.96-6.88 (m,
2H), 6.22 (d, J=2.3 Hz, 1H), 6.02 (s, 2H), 1.43-1.40 (m, 2H), 1.37
(s, 9H), 1.06-1.02 (m, 2H).
Example 89
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(1,3-dihydroxypropan-2-yl)--
1H-indol-5-yl)cyclopropanecarboxamide
##STR00743##
[0609]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(1,3-dihydroxypropan-
-2-yl)indolin-5-yl)cyclopropanecarboxamide
[0610]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butylindolin-5-yl)cyclopropa-
necarboxamide (50 mg, 0.13 mmol) was dissolved in dichloroethane
(0.20 mL) and 2,2-dimethyl-1,3-dioxan-5-one (0.20 mL).
Trifluoroacetic acid was added (0.039 mL) and the resulting
solution was allowed to stir for 20 minutes. Sodium
triacetoxyborohydride was added (55 mg, 0.26 mmol) and the reaction
mixture was stirred for 30 minutes. The crude reaction mixture was
then evaporated to dryness, dissolved in N,N-dimethylformamide and
purified by preparative HPLC using a gradient of 0-99% acetonitrile
in water containing 0.05% trifluoroacetic acid.
##STR00744##
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(1,3-dihydroxypropan-2-yl)-
-1M-indol-5-yl)cyclopropanecarboxamide
[0611]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(1,3-dihydroxypropan-
-2-yl)indolin-5-yl)cyclopropanecarboxamide (40.3 mg, 0.0711 mmol as
the trifluoracetic acid salt) was dissolved in toluene (1 mL). To
the resulting solution was added
2,3,5,6-tetrachlorocyclohexa-2,5-diene-1,4-dione (35 mg, 0.14
mmol). The resulting suspension was heated at 100.degree. C. in an
oil bath for 10 minutes. The crude product was then evaporated to
dryness, dissolved in a 1 mL of N,N-dimethylformamide and purified
by purified by preparative HPLC using a gradient of 0-99%
acetonitrile in water containing 0.05% trifluoroacetic acid to
yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(1,3-dihydroxypropan-2-yl)-
-1H-indol-5-yl)cyclopropanecarboxamide. ESI-MS m/z calc. 450.2,
found 451.5 (M+1)*. Retention time of 1.59 minutes.
Example 90
N-(7-(Aminomethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]-dioxol-5-y-
l)cyclopropanecarboxamide
##STR00745##
[0612]
N-(7-(Aminomethyl-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]dioxo-
l-5 yl)cyclopropanecarboxamide
[0613]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-7-cyano-1H-indol-5-yl)-
cyclopropanecarboxamide (375 mg, 0.934 mmol) was dissolved in 35 mL
of ethyl acetate. The solution was recirculated through a
continuous flow hydrogenation reactor containing 10% palladium on
carbon at 100.degree. C. under 100 bar of hydrogen for 8 h. The
crude product was then evaporated to dryness and purified on 12 g
of silica gel utilizing a gradient of 0-100% ethyl acetate
(containing 0.5% triethylamine) in hexanes to yield
N-(7-(aminomethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]-dioxol-5--
yl)-cyclopropanecarboxamide (121 mg, 32%). ESI-MS m/z calc. 405.2,
found 406.5 (M+I). Retention time of 1.48 minutes.
Example 91
5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butyl-1H-in-
dole-7-carboxamide
##STR00746##
[0614]
5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-buty-
l-1H-indole-7-carboxamide
[0615]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-7-cyano-1H-indol-5-yl)-
-cyclopropanecarboxamide (45 mg, 0.11 mmol) was suspended in a
mixture of methanol (1.8 mL), 30% aqueous hydrogen peroxide (0.14
mL, 4.4 mmol) and 10% aqueous sodium hydroxide (0.150 mL). The
resulting suspension was stirred for 72 h at room temperature. The
hydrogen peroxide was then quenched with sodium sulfite. The
reaction mixture was diluted with 0.5 mL of N,N-dimethylformamide,
filtered, and purified by preparative HPLC using a gradient of
0-99% acetonitrile in water containing 0.05% trifluoroacetic acid
to yield
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropane-carboxamido)-2-tert-butyl-1H--
indole-7-carboxamide. ESI-MS m/z calc. 419.2, found 420.3
(M+I).sup.+. Retention time of 1.74 minutes.
Example 92
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-7-(methylsulfonamido-methyl)--
1H-indol-5-yl)cylopropanecarboxamide
##STR00747##
[0616]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-7-(methylsulfonamidome-
thyl)-1H-indol-5-yl)cyclopropanecarboxamide
[0617]
N-(7-(Aminomethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d[1,3]dioxo-
l-5-yl)cyclopropanecarboxamide (20 mg, 0.049 mmol) was dissolved in
DMF (0.5 mL) containing triethylamine (20.6 .mu.L, 0.147 mmol) and
a magnetic stir bar. Methanesulfonyl chloride (4.2 .mu.L, 0.054
mmol) was then added to the reaction mixture. The reaction mixture
was allowed to stir for 12 h at room temperature. The crude product
was purified by preparative HPLC using a gradient of 0-99%
acetonitrile in water containing 0.05% trifluoroacetic acid to
yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-7-(methylsulfonamidomethyl)--
1H-indol-5-yl)cyclopropanecarboxamide. ESI-MS m/z calc. 483.2,
found 484.3 (M+l). Retention time of 1.84 minutes.
Example 93
N-(7-(Acetamidomethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]-dioxol-
-5-yl)cyclopropanecarboxamide
##STR00748##
[0619]
N-(7-(Aminomethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]diox-
ol-5-yl)cyclopropanecarboxamide (20 mg, 0.049 mmol) was dissolved
in DMF (0.5 mL) containing triethylamine (20.6 .mu.L, 0.147 mmol)
and a magnetic stir bar. Acetyl chloride (4.2 .mu.L, 0.054 mmol)
was then added to the reaction mixture. The reaction mixture was
allowed to stir for 16 h at room temperature. The crude product was
purified by preparative HPLC using a gradient of 0-99% acetonitrile
in water containing 0.05% trifluoroacetic acid to yield
N-(7-(acetamidomethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]dioxol-
-5-yl)cyclopropanecarboxamide. ESI-MS m/z calc. 447.2, found 448.3
(M+1)+. Retention time of 1.76 minutes.
Example 94
N-(1-Acetyl-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]dioxol-5-yl)-cyclo-
propanecarboxamide
##STR00749##
[0621] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)cyclopropaneca-
rboxamide (120 mg, 0.31 mmol) in anhydrous DMF-THF (3.3 mL, 1:9)
was added NaH (60% in mineral oil, 49 mg, 1.2 mmol) at room
temperature. After 30 min under N.sub.2, the suspension was cooled
down to -15.degree. C. and a solution of acetyl chloride (1.1 eq.)
in DMF (0.5 mL) was added dropwise. The reaction mixture was
stirred for 30 min at -15.degree. C. then for 6 h at room
temperature. Water (0.5 mL) was added at 0.degree. C., solvent was
removed, and the residue was diluted with MeOH, filtrated and
purified by preparative HPLC to give
N-(1-acetyl-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]dioxol-5-yl)cyclo-
propanecarboxamide. .sup.1H NMR (400 MHz, DMSO) .delta. 8.9 (s,
1H), 7.74 (d, J=2.1 Hz, 1H), 7.54 (d, J=9.0 Hz, 1H), 7.28 (dd,
J1=2.1 Hz; J2=9.0 Hz, 1H), 7.01 (d, J=1.5 Hz, 1H), 6.93 (dd, J1=1.7
Hz, J2=8.0 Hz, 1H), 6.89 (d, J=8.0 Hz, 1H), 6.54 (bs, 1H), 6.02 (s,
2H), 2.80 (s, 3H), 1.42-1.40 (m, 11H), 1.06-1.05 (m, 2H). MS (ESI)
m/e (M+H.sup.+) 419.3.
Example 95
N-(1-(2-Acetamidoethyl)-2-tert-butyl-6-fluoro-1-indol-5-yl)-1-(2,2-difluor-
obenzo[d](1,3]dioxol-5-yl)cyclopropanecarboxamide
##STR00750## ##STR00751##
[0622]
N-(1-(2-Aminoethyl)-2-tert-butyl-6-fluoro-1H-indol-5-yl)-1-(2,2-dif-
luorobenzo-1[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0623] To a solution of tert-butyl
2-(2-tert-butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropaneca-
rboxamido)-6-fluoro-1H-indol-1-yl)ethylcarbamate (620 mg, 1.08
mmol) in CH.sub.2Cl.sub.2 (8 mL) was added TFA (2 mL). The reaction
was stirred at room temperature for 1.5 h before being neutralized
with solid NaHCO.sub.3. The solution was partitioned between
H.sub.2O and CH.sub.2Cl.sub.2. The organic layer was dried over
MgSO.sub.4, filtered and concentrated to yield the product as a
cream colored solid (365 mg, 71%). .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 8.38 (s, 1H), 7.87 (br s, 3H, NH.sub.3.sup.+), 7.52 (s,
1H), 7.45-7.38 (m, 3H), 7.32 (dd, J=8.3, 1.5 Hz, 1H), 6.21 (s, 1H),
4.46 (m, 2H), 3.02 (m, 2H), 1.46 (m, 2H), 1.41 (s, 9H), 1.14 (m,
2H). HPLC ret. time 1.66 min, 10-99% CH.sub.3CN, 3 min run; ESI-MS
474.4 m/z (M+H.sup.+).
##STR00752##
N-(1-(2-Acetamidoethyl)-2-tert-butyl-6-fluoro-1H-indol-5-yl)-1-(2,2-diflu-
orobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0624] To a solution of
N-(1-(2-aminoethyl)-2-tert-butyl-6-fluoro-1H-indol-5-yl)-1-(2,2-difluorob-
enzo[d][1,3]dioxol-5-yl)cyclopropane-carboxamide (47 mg, 0.10 mmol)
and Et.sub.3N (28 .mu.L, 0.20 mmol) in DMF (1 mL) was added acetyl
chloride (7.1 .mu.L, 0.10 mmol). The mixture was stirred at room
temperature for 1 h before being filtered and purified by reverse
phase HPLC (10-99% CH.sub.3CN/H.sub.2O) to yield
N-(1-(2-acetamidoethyl)-2-tert-butyl-6-fluoro-1H-indol-5-yl)-1-(2,2-diflu-
orobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide. .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 8.35 (s, 1H), 8.15 (t, J=5.9 Hz, 1H),
7.53 (s, 1H), 7.43-7.31 (m, 4H), 6.17 (s, 1H), 4.22 (m, 2H), 3.30
(m, 2H), 1.85 (s, 3H), 1.47 (m, 2H), 1.41 (s, 9H), 1.13 (m, 2H).
HPLC ret. time 2.06 min, 10-99% CH.sub.3CN, 3 min run; ESI-MS 516.4
m/z (M+H.sup.+).
Example 96
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(2-hydroxy-3-methoxy-propyl-
)-1H-indol-5-yl)cyclopropanecarboxamide
##STR00753##
[0626]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)cyclopro-
panecarboxamide (320 mg, 0.84 mmol) was dissolved in a mixture
composed of anhydrous DMF (0.5 mL) and anhydrous THF (5 mL) under
N.sub.2. NaH (60% in mineral oil, 120 mg, 3.0 mmol) was added at
room temperature. After 30 min of stirring, the reaction mixture
was cooled to -15.degree. C. before a solution of epichlorohydrin
(79 .mu.L, 1.0 mmol) in anhydrous DMF (1 mL) was added dropwise.
The reaction mixture was stirred for 15 min at -15.degree. C., then
for 8 h at room temperature. MeOH (1 mL) was added and the mixture
was heated for 10 min at 105.degree. C. in the microwave oven. The
mixture was cooled, filtered and purified by preparative HPLC to
give
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(2-hydroxy-3-metho-
xy-propyl)-1H-indol-5-yl)cyclopropanecarboxamide. .sup.1H NMR (400
MHz, DMSO-d6) .delta. 8.44 (s, 18), 7.59 (d, J=1.9 Hz, 1H), 7.31
(d, J=8.9 Hz, 1H), 7.03 (dd, J=8.7, 1.9 Hz, 2H), 6.95 (dd, J=8.0,
1.7 Hz, 1H), 6.90 (d, J=8.0 Hz, 1H), 6.16 (s, 1H), 6.03 (s, 2H),
4.33 (dd, J=15.0, 4.0 Hz, 1H), 4.19 (dd, J=15.0, 8.1 Hz, 1H), 4.02
(ddd, J=8.7, 4.8 Hz, 1H), 3.41-3.32 (m, 2H), 3.30 (s, 3H), 1.41 (s,
9H), 1.41-1.38 (m, 2H), 1.03 (dd, J=6.7, 4.0 Hz, 2H). MS (ESI) m/e
(M+H.sup.+) 465.0.
Example 97
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(2-hydroxy-3-(methyl-amino)-
propyl)-1H-indol-5-yl)cyclopropanecarboxamide
##STR00754##
[0628]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)cyclopro-
panecarboxamide (320 mg, 0.84 mmol) was dissolved in a mixture
composed of anhydrous DMF (0.5 mL) and anhydrous THF (5 mL) under
N.sub.2. NaH (60% in mineral oil, 120 mg, 3.0 mmol) was added at
room temperature. After 30 min of stirring, the reaction mixture
was cooled to -15.degree. C. before a solution of epichlorohydrin
(79 .mu.L, 1.0 mmol) in anhydrous DMF (1 mL) was added dropwise.
The reaction mixture was stirred for 15 min at -15.degree. C., then
for 8 h at room temperature. MeNH.sub.2 (2.0 M in MeOH, 1.0 mL) was
added and the mixture was heated for 10 min at 105.degree. C. in
the microwave oven. The mixture was cooled, filtered and purified
by preparative HPLC to give
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(2-hydroxy-3-(methylamino)-
propyl)-1H-indol-5-yl)cyclopropanecarboxamide. .sup.1H NMR (400
MHz, DMSO-d6) .delta. 8.50 (s, 1H), 7.60-7.59 (m, 1H), 7.35 (dd,
J=14.3, 8.9 Hz, 1H), 7.10 (d, J=8.8 Hz, 1H), 1H), 6.94 (dd, J=8.0,
1.6 Hz, 1H), 6.91 (d, J=7.9 Hz, 1H), 6.20 (d, J=2.3 Hz, 1H), 6.03
(s, 2H), 2.82 (d, J=4.7 Hz, 1H), 2.72 (d, J=4.7 Hz, 1H), 2.55 (dd,
J=5.2, 5.2 Hz, 1H), 2.50 (s, 3H), 1.43 (s, 9H), 1.39 (dd, J=6.4,
3.7 Hz, 2H), 1.04 (dd, J=6.5, 3.9 Hz, 2H). MS (ESI) m/e (M+H.sup.+)
464.0.
Example 98
(S)--N-(1-(3-Amino-2-hydroxypropyl)-2-tert-butyl-1H-indol-5-yl)-1-(2,2-dif-
luorobenzo[d][1,3])dioxol-5-yl)cyclopropanecarboxamide
##STR00755##
[0629]
(R)-3-(2-tert-Butyl-1-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cycl-
opropanecarbox-amido)-1H-indol-1-yl)-2-hydroxypropyl-4-methylbenzenesulfon-
ate
[0630] To a stirred solution of
(R)--N-(2-tert-butyl-1-(2,3-dihydroxypropyl)-1H-indol-5-yl)-1-(2,2-difluo-
ro-benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide (3.0 g, 6.1
mmol) in dichloromethane (20 mL) was added triethylamine (2 mL) and
para-toluenesulfonylchloride (1.3 g, 7.0 mmol). After 18 hours, the
reaction mixture was partitioned between 10 mL of water and 10 mL
of ethyl acetate. The organic layer was dried over magnesium
sulfate, filtered and evaporated. The residue was purified using
column chromatography on silica gel (0-60% ethyl acetate/hexane)
providing (R)-3-(2-tert-butyl-5-(1-(2,2-difluorobenzo[d)
1,3]-dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-1-yl)-2-hydroxypropyl--
4-methylbenzenesulfonate (3.21 g, 86%). LC/MS (M+1)=641.2. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.77 (d, 2H, J=16 Hz), 7.55 (d,
1H, J=2 Hz), 7.35 (d, 2H, J=16 Hz), 7.31 (m, 3H), 6.96 (s, 1H),
6.94 (dd, 1H, J=2, 8 Hz), 6.22 (s, 1H), 4.33 (m, 1H), 4.31 (dd, 1H,
J=6, 15 Hz), 4.28 (dd, 1H, J=11, 15 Hz), 4.18 (m, 1H), 3.40 (dd,
1H, J=3, 6 Hz), 3.36 (dd, 11, J=3, 6 Hz), 2.46 (s, 3H), 2.40 (br s,
1H), 1.74 (m, 2H), 1.40 (s, 9H), 1.11 (m, 2H).
##STR00756##
(R)--N-(1-(3-Azido-2-hydroxypropyl)-2-tert-butyl-1H-indol-5-yl)-1-(2,2-di-
fluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0631] To a stirred solution
(R)-3-(2-tert-butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropa-
necarboxamido)-1H-indol-1-yl)-2-hydroxypropyl-4-methylbenzenesulfonate
(3.2 g, 5.0 mmol) in DMF (6 mL) was added sodium azide (2.0 g, 30
mmol). The reaction was heated at 80.degree. C. for 2 h. The
mixture was partitioned between 20 mL ethyl acetate and 20 mL
water. The layers were separated and the organic layer was
evaporated. The residue was purified using column chromatography
(0-85% ethyl acetate/hexane) to give
(R)--N--(I-(3-azido-2-hydroxypropyl)-2-tert-butyl-1H-indol-5-yl)-1-(2,2-d-
ifluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide (2.48 g).
LC/MS (M+1)=512.5. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.55
(d, 1H, J=2 Hz), 7.31 (m, 3H), 6.96 (s, 1H), 6.94 (dd, 1H, J=2, 8
Hz), 6.22 (s, 1H), 4.33 (m, 1H), 4.31 (dd, 1H, J=6, 15 Hz), 4.28
(dd, 1H, J=11, 15 Hz), 4.18 (m, 1H), 3.40 (dd, 1H, J=3, 6 Hz), 3.36
(dd, 1H, J=3, 6 Hz), 2.40 (br s, 1H), 1.74 (m, 2H), 1.40 (s, 9H),
1.11 (m, 2H).
##STR00757##
(S)--N-(1-(3-Amino-2-hydroxypropyl)-2-tert-butyl-1H-indol-5-yl)-1-(2,2-di-
fluoro-benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0632] To a stirred solution
(R)--N-(1-(3-azido-2-hydroxypropyl)-2-tert-butyl-1H-indol-5-yl)-1-(2,2-di-
fluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide (2.4 g, 4.0
mmol) in MeOH (25 mL) was added 5% Pd/C (2.4 g) under a Hydrogen
gas filled balloon. After 18 h, the reaction mixture was filtered
through celite and rinsed with 300 mL ethyl acetate. The organic
layer was washed with 1 N HCl and evaporated to give
(S)--N-(1-(3-amino-2-hydroxypropyl)-2-tert-butyl-1H-indol-5-yl)-1-(2,2-di-
fluoro-benzo[d][1,3]-dioxol-5-yl)cyclopropane-carboxamide (1.37 g).
MS (M+1)=486.5.
Example 99
(S)-Methyl
3-(2-tert-butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cycl-
opropanecarboxamido)-1H-indol-1-yl)-2-hydroxypropylcarbamate
##STR00758##
[0634] To a stirred solution
(R)--N-(1-(3-amino-2-hydroxypropyl)-2-tert-butyl-1H-indol-5-yl)-1-(2,2-di-
fluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide (0.10 g,
0.20 mmol) in methanol (1 mL) was added 2 drops of triethylamine
and methylchloroformyl chloride (0.020 mL, 0.25 mmol). After 30
min, the reaction mixture was filtered and purified using reverse
phase HPLC providing (S)-methyl
3-(2-tert-butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclo-propanec-
arboxamido)-1H-indol-1-yl)-2-hydroxypropylcarbamate. The retention
time on a three minute run is 1.40 min. LC/MS (M+1)=544.3. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.52 (d, 1H, J=2 Hz), 7.30 (dd,
1H, J=2, 8 Hz), 7.28 (m, 1H), 7.22 (d, 1H, J=8 Hz), 7.14 (d, 1H,
J=8 Hz), 7.04 (br s, 1H), 6.97 (dd, 1H, J=2, 8 Hz), 6.24 (s, 1H),
5.19 (1H, br s), 4.31 (dd, 1H, J=6, 15 Hz), 4.28 (dd, 1H, J=11, 15
Hz), 4.18 (m, 1H), 3.70 (s, 3H), 3.40 (dd, 1H, J=3, 6 Hz), 3.36
(dd, 1H, J=3.6 Hz), 3.26 (m, 1H), 1.74 (m, 2H), 1.40 (s, 9H), 1.11
(m, 2H).
Example 100
4-(5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butyl-1H-
-indol-1-yl)butanoic acid
##STR00759##
[0635]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butylindolin-5-yl)cyclopropa-
necarboxamide
[0636] To 4 solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1H-indol-5-yl)cyclopropaneca-
rboxamide (851 mg, 2.26 mmol) in acetic acid (60 mL) was added
NaBH.sub.3CN (309 mg, 4.91 mmol) at 0.degree. C. The reaction
mixture was stirred for 5 min at room temperature after which no
starting material could be detected by LCMS. The solvent was
evaporated under reduced pressure and the residue was purified by
column chromatography on silica gel (5-40% ethyl acetate/hexanes)
to give
3-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butylindolin-5-yl)cyclopropanecarb-
oxamide (760 mg, 89%).
##STR00760##
4-(5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butylin-
dolin-1-yl)butanoic acid
[0637] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butylindolin-5-yl)cyclopropanecarb-
oxamide (350 mg, 0.93 mmol, 1 eq) in anhydrous methanol (6.5 mL)
and AcOH (65 .mu.L) was added 4-oxobutanoic acid (15% in water, 710
mg, 1.0 mmol) at room temperature. After 20 min of stirring,
NaBH.sub.3CN (130 mg, 2.0 mmol) was added in one portion and the
reaction mixture was stirred for another 4 h at room temperature.
The reaction mixture was quenched by addition of AcOH (0.5 mL) at
0.degree. C. and the solvent was removed under reduced pressure.
The residue was purified by column chromatography on silica gel
(5-75% ethyl acetate/hexanes) to give
4-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butylin-
dolin-1-yl)butanoic acid (130 mg, 30%).
##STR00761##
4-(5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butyl-1-
H-indol-1-yl)butanoic acid
[0638]
4-(5-(1-(Benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-b-
utylindolin-1-yl)butanoic acid (130 mg, 0.28 mmol) was taken up in
a mixture of acetonitrile-H.sub.2O-TFA. The solvent was removed
under reduced pressure and the residue obtained was dissolved in
CDCl.sub.3. After a brief exposition to daylight (5-10 min), the
solution turned purple. The mixture was stirred open to the
atmosphere at room temperature until complete disappearance of the
starting material (8 h). Solvent was removed under reduced pressure
and the residue was purified by reverse phrase HPLC to give
4-(5-(l-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butyl-1-
H-indol-1-yl)butanoic acid. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.52 (d, J=1.9 Hz, 1H), 7.18 (d, J=2.1 Hz, 1H), 7.16 (s,
1H), 7.03 (dd, J=9.4, 1.9 Hz, 1H), 7.00-6.98 (m, 2H), 6.85 (d,
J=7.9 Hz, 1H), 6.16 (s, 1H), 6.02 (s, 2H), 4.29-4.24 (m. 2H), 2.48
(dd, J=6.9, 6.9 Hz, 2H), 2.12-2.04 (m, 2H), 1.69 (dd, J=6.8, 3.7
Hz, 2H), 1.43 (s, 9H), 1.09 (dd, J=6.8, 3.7 Hz, 2H). MS (Esr) m/e
(M+H+) 463.0.
Example 101
1-(Benzo[d]][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(4-(2-hydroxyethyl-amino)--
4-oxobutyl)-1H-indol-5-yl)cyclopropanecarboxamide
##STR00762##
[0640] To a solution of
4-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butyl-1-
H-indol-1-yl)butanoic acid (10 mg) in anhydrous DMF (0.25 mL) were
successively added Et.sub.3N (9.5 mL, 0.069 mmol) and HBTU (8.2 mg,
0.022 mmol). After stirring for 10 min at 60.degree. C.,
ethanolamine (1.3 .mu.L, 0.022 mmol) was added, and the mixture was
stirred for another 4 h at 60.degree. C.
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(4-(2-hydroxyethyl-amino)--
4-oxobutyl)-1H-indol-5-yl)cyclopropanecarboxamide (5.8 mg, 64%) was
obtained after purification by preparative HPLC. MS (ESI) m/e
(M+H.sup.+) 506.0.
Example 102
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(2-(dimethylamino)-2-oxoeth-
yl)-1H-indol-5-yl)cyclopropanecarboxamide
##STR00763##
[0642] To a solution of 1-(benzo
d[1,3]dioxol-5-yl)-N-(2-tert-butylindolin-5-yl)cyclopropanecarboxamide
(62 mg, 0.16 mmol) in anhydrous DMF (0.11 mL) and THF (1 mL) was
added NaH (60% in mineral oil, 21 mg, 0.51 mmol) at room
temperature under N.sub.2. After 30 min of stirring, the reaction
mixture was cooled to 0.degree. C. and
2-chloro-N,N-dimethylacetamide (11 mL, 0.14 mmol) was added. The
reaction mixture was stirred for 5 min at 0.degree. C. and then for
10 h at room temperature. The mixture was purified by preparative
HPLC and the resultant solid was dissolved in DMF (0.6 mL) in the
presence of Pd--C(10 mg). The mixture was stirred open to the
atmosphere overnight at room temperature. The reaction mixture was
filtrated and purified by preparative HPLC providing
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(2-(dimethylamino)-2-oxoet-
hyl)-1H-indol-5-yl)cyclopropanecarboxamide. MS (ESI) m/e
(M+H.sup.+) 462.0.
Example 103
3-(2-tert-Butyl-5-(1-(2,2difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarb-
oxamido)-1H-indol-1-yl)propanoic acid
##STR00764##
[0643]
N-(2-tert-Butyl-1-(2-chloroethyl)indolin-5-yl)-1-(2,2-difluorobenzo-
[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0644] To a solution of
N-(2-tert-butyl-1-(2-cyanoethyl)indolin-5-yl)-1-(2,2-difluorobenzo[d][1,3-
]dioxol-5-yl)cyclopropanecarboxamide (71 mg, 0.17 mmol) in
anhydrous dichloromethane (1 mL) was added chloroacetaldehyde (53
.mu.L, 0.41 mmol) at room temperature under N.sub.2. After 20 min
of stirring, NaBH(OAc).sub.3 (90 mg, 0.42 mmol) was added in two
portions. The reaction mixture was stirred overnight at room
temperature. The product was purified by column chromatography on
silica gel (2-15% ethyl acetate/hexanes) providing
N-(2-tert-butyl-1-(2-chloroethyl)indolin-5-yl)-1-(2,2-difluorobenzo[d][1,-
3]dioxol-5-yl)cyclopropanecarboxamide (51 mg, 63%).
##STR00765##
N-(2-tert-Butyl-1-(2-cyanoethyl)indolin-5-yl)-1-(2,2-difluorobenzo[d][1,3-
]dioxol-5-yl)cyclopropanecarboxamide
[0645]
N-(2-tert-butyl-1-(2-chloroethyl)indolin-5-yl)-1-(22-difluorobenzo[-
d][1,3]dioxol-5-yl)cyclopropanecarboxamide (51 mg), NaCN (16 mg,
0.32 mmol) and KI (cat) in EtOH (0.6 mL) and water (0.3 mL) were
combined and heated at 110.degree. C. for 30 min in the microwave.
The solvent was removed under reduced pressure and the residue was
purified by column chromatography on silica gel (2-15% ethyl
acetate/hexanes) providing
N-(2-tert-butyl-1-(2-cyanoethyl)indolin-5-yl)-1-(2,2-difluorobenzo[d][1,3-
]dioxol-5-yl)cyclopropanecarboxamide (24 mg, 48%).
##STR00766##
3-(2-tert-Butyl-5-(1-(2,2-difluorobenzo[d]1,3]dioxol-5-yl)cyclo-propaneca-
rboxamido)-1H-indol-1-yl)propanoic acid
[0646]
N-(2-tert-butyl-1-(2-cyanoethyl)indolin-5-yl)-1-(2,2-difluorobenzo[-
d][1,3]dioxol-5-yl)cyclopropane-carboxamide (24 mg, 0.050 mmol) was
taken up in 50% aq. KOH (0.5 mL) and 1,4-dioxane (1 mL). The
mixture was heated at 125.degree. C. for 2 h. The solvent was
removed and the residue was purified by preparative HPLC. The
residue was dissolved in CDCl.sub.3 (1 mL) then briefly exposed to
daylight. The purple solution that formed was stirred until
complete disappearance of the starting material (1 h). The solvent
was removed under reduced pressure and the residue was purified by
preparative HPLC providing
3-(2-tert-butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclo-propanec-
arboxamido)-1H-indol-1-yl)propanoic acid. MS (ESI) m/e (M+H
485.0.
Example 104
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoro-1-(2-hydroxy-ethyl)--
1H-indol-5-cyclopropanecarboxamide
##STR00767##
[0648] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoroindolin-5-yl)cyclopr-
opanecarboxamide (340 mg, 0.86 mmol) in anhydrous MeOH (5.7 mL)
containing 1% of acetic acid was added glyoxal 40% in water (0.60
mL, 5.2 mmol) at room temperature under N.sub.2. After 20 min of
stirring, NaBH.sub.3CN (120 mg, 1.9 mmol) was added in one portion
and the reaction mixture was stirred overnight at room temperature.
The solvent was removed under reduced pressure and the residue
obtained was purified by column chromatography on silica gel
(10-40% ethyl acetate/hexanes) providing a pale yellow oil which
was treated with 50/50 CH.sub.3CN--H.sub.2O containing 0.05% TFA
and CDCl.sub.3. Solvent was removed under reduced pressure and the
residue was purified by column chromatography on silica gel (20-35%
ethyl acetate/hexanes) to give
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoro-1-(2-hydroxyethyl)--
1H-indol-5-yl)cyclopropanecarboxamide. .sup.1H NMR (400 MHz,
CDCl.sub.3) 88.02 (d, J=7.7 Hz, 11), 7.30 (d, J=2.1 Hz, 1H); 6.93
(dd, J=1.6, 7.9 Hz, 1H), 6.90 (d, J=1.6 Hz, 11), 6.90 (d, J=1.6 Hz,
1H), 6.78 (d, J=7.9 Hz, 11H), 6.08 (s, 1H), 5.92 (s, 2H), 4.21 (dd,
J=6.9, 6.9 Hz, 2H), 3.68 (m, 2H), 2.28 (s, 1H), 1.60 (dd, J=3.7,
6.7 Hz, 2H), 1.35-1.32 (m, 9H), 1.04 (dd, J=3.7, 6.8 Hz, 2H). MS
(ESI) m/e (M+H.sup.+) 439.0.
Example 105
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoro-1-(3-hydroxy-propyl)-
-1H-indol-5-yl)cyclopropanecarboxamide
##STR00768##
[0649] 3-(Benzyloxy)propanal
[0650] To a suspension of PCC (606 mg, 2.82 mmol) in anhydrous
dichloromethane (8 mL) at room temperature under N.sub.2 was added
a solution of 3-benzyloxy-1-propanol (310 mg, 1.88 mmol) in
anhydrous dichloromethane. The reaction mixture was stirred
overnight at room temperature, filtrated through Celite, and
concentrated. The residue was purified by column chromatography on
silica gel (1-10% ethyl acetate/hexanes) to give
3-(benzyloxy)propanal (243 mg, 79%).
##STR00769##
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoro-1-(3-hydroxypropyl)-
-1H-indol-5-yl)cyclopropanecarboxamide
[0651] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoroindolin-5-yl)cyclopr-
opanecarboxamide (160 mg, 0.50 mmol) in anhydrous dichloromethane
(3.4 mL) was added 3-(benzyloxy)propanal (160 mg, 0.98 mmol) at
room temperature. After 10 min of stirring, NaBH(OAc).sub.3 (140
mg, 0.65 mmol) was added in one portion and the reaction mixture
was stirred for 4 h at room temperature. The solvent was removed
under reduced pressure and the residue was taken-up in a mixture of
50/50 CH.sub.3CN--H.sub.2O containing 0.05% TFA. The mixture was
concentrated to dryness and the residue was dissolved in CDCl.sub.3
(5 mL) and briefly exposed to daylight. The purple solution was
stirred open to the atmosphere at room temperature for 2 h. The
solvent was removed under reduced pressure and the residue was
treated with Pd--C(10 mg) in MeOH (2 mL) under 1 atm of H.sub.2 for
2 h. The catalyst was filtered through Celite and the solvent was
removed under reduced pressure. The residue was purified by
preparative TLC 30% ethyl acetate/hexanes to provide
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoro-1-(3-hydroxypropyl)-
-1H-indol-5-yl)cyclopropanecarboxamide (18 mg, 8% from
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoroindolin-5-yl)cyclopr-
opane-carboxamide). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.11
(d, J=7.8 Hz, 1H), 7.31 (d, J=2.2 Hz, 1H), 6.94 (dd, J=7.9, 1.7 Hz,
1H), 6.91 (d, J=1.6 Hz, 1H), 6.85 (d, J=11.7 Hz, 1H), 6.79 (d,
J=7.9 Hz, 1H), 6.10 (s, 1H), 5.94 (s, 2H), 4.25-4.21 (m, 2H), 3.70
(dd, J=5.7, 5.7 Hz, 2H), 1.93-1.86 (m, 2H), 1.61 (dd, J=6.8, 3.7
Hz, 2H), 1.35 (s, 9H), 1.04 (dd, J=6.8, 3.7 Hz, 2H). MS (ESI) m/e
(M+H.sup.+) 453.0.
Example 106
N-(1-(2-Acetamidoethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]-dioxo-
l-5-yl)cyclopropanecarboxamide
##STR00770##
[0652]
N-(1-(2-azidoethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]dio-
xol-5-yl)-cyclopropanecarboxamide
[0653] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butylindolin-5-yl)cyclopropane-car-
boxamide (73 mg, 0.19 mmol) in anhydrous dichloromethane (1.2 mL)
was added chloroacetaldehyde (60 .mu.L, 0.24 mmol) at room
temperature. After 10 min of stirring, NaBH(OAc).sub.3 (52 mg, 0.24
mmol) was added in one portion and the reaction mixture was stirred
for another 30 min at room temperature. The solvent was removed
under reduced pressure and the residue was purified by preparative
HPLC to give the indoline, which oxidized to the corresponding
indole when taken-up in CDCl.sub.3. The resulting indole was
treated with NaN.sub.3 (58 mg, 0.89 mmol) and NaI (cat) in
anhydrous DMF (0.8 mL) for 2 h at 85.degree. C. The reaction
mixture was purified by preparative HPLC providing
N-(1-(2-azidoethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]dioxol-5--
yl)cyclopropanecarboxamide (15 mg, 18% from
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butylindolin-5-yl)cyclopropane-car-
boxamide).
##STR00771##
N-(1-(2-Acetamidoethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]-diox-
ol-5-yl)cyclopropanecarboxamide
[0654] A solution of
N-(1-(2-azidoethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]dioxol-5--
yl)cyclopropanecarboxamide (13 mg, 0.029 mmol) in MeOH-AcOH (0.2
mL, 99:1) in the presence of Pd--C(2 mg) was stirred at room
temperature under 1 atm of H.sub.2 for 2 h, filtered through
Celite, and concentrated under reduced pressure. The crude product
was treated with AcCl (0.05 mL) and Et.sub.3N (0.05 mL) in
anhydrous THF (0.2 mL) at 0.degree. C. for 30 min and then 1 h at
room temperature. The mixture was purified by preparative HPLC
providing
N-(1-(2-acetamidoethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1,3]-diox-
ol-5-yl)cyclopropanecarboxamide. MS (ESI) m/e (M+H.sup.+)
462.0.
Example 107
N-(2-tert-Butyl-1-(3-cyano-2-hydroxypropyl)-1H-indol-5-yl)-1-(2,2-difluoro-
benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
##STR00772##
[0655]
3-(2-tert-Butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopro-
panecarboxamido)-1H-indol-1-yl)-2-hydroxypropyl-4-methylbenzenesulfonate
[0656] To a solution of
N-(2-tert-butyl-1-(2,3-dihydroxypropyl)-1H-indol-5-yl)-1-(2,2-difluoroben-
zo[d][1,3]-dioxol-5-yl)cyclopropanecarboxamide (172 mg, 0.35 mmol)
in anhydrous dichloromethane (1.4 mL) at 0.degree. C. in the
presence of Et.sub.3N (56 .mu.L, 0.40 mmol) was added TsCl (71 mg,
0.37 mmol). The reaction mixture was stirred for 2 h at room
temperature before being cooled to 0.degree. C. and another portion
of TsCl (71 mg, 0.37 mmol) was added. After 1 h of stirring at room
temperature, the mixture was purified by column chromatography on
silica gel (10-30% ethyl acetate/hexanes) providing
3-(2-tert-butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropaneca-
rboxamido)-1H-indol-1-yl)-2-hydroxypropyl-4-methylbenzene-sulfonate
(146 mg, 64%).
##STR00773##
N-(2-tert-Butyl-1-(3-cyano-2-hydroxypropyl)-1H-indol-5-yl)-1-(2,2-difluor-
obenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0657]
N-(2-tert-Butyl-1-(3-cyano-2-hydroxypropyl)-1H-indol-5-yl)-1-(2,2-d-
ifluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamide (145 rag,
0.226 mmol) was treated with powdered NaCN (34 mg, 0.69 mmol) in
anhydrous DMF (1.5 mL) at 85.degree. C. for 2 h. The reaction
mixture was cooled down to room temperature before it was diluted
with dichloromethane (10 mL) and aq. sat. NaHCO.sub.3 (10 mL). The
organic phase was separated and the aqueous phase was extracted
with dichloromethane (2.times.10 mL). The organic phases were
combined, washed with brine, dried with sodium sulfate, filtered
then concentrated. The residue was purified by column
chromatography on silica gel (25-55% ethyl acetate/hexanes)
providing
N-(2-tert-butyl-1-(3-cyano-2-hydroxypropyl)-1H-indol-5-yl)-1-(2,2-difluor-
obenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide (89 mg, 79%).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.43 (d, J=1.9 Hz, 1H),
7.20-7.16 (m, 2H), 7.08 (d, J=8.8 Hz, 1H), 7.04 (d, J=8.2 Hz, 1H),
6.94 (s, 1H), 6.88 (dd, J=8.7, 2.0 Hz, 1H), 6.16 (s, 1H), 4.32-4.19
(m, 3H), 2.83 (s, 1H), 2.40 (dd, J=5.2, 5.2 Hz, 2H), 1.62 (dd,
J=6.6, 3.6 Hz, 2H), 1.35 (s, 9H), 1.04 (dd, J=6.9, 3.9 Hz, 2H). MS
(ESI) m/e (M+H.sup.+) 496.0.
Example 108
N-(2-tert-Butyl-1-(2-hydroxy-3-(2H-tetrazol-5-yl)propyl)-1H-indol-5-yl)-1--
(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
##STR00774##
[0659] To a solution of N-(2-tert-butyl-1
(3-cyano-2-hydroxypropyl)-1H-indol-5-yl)-1-(2,2-difluorobenzo[d][1,3]diox-
ol-5-yl)cyclopropanecarboxamide (27 mg, 0.054 mmol) in anhydrous
DMF (1.2 mL) were successively added NH.sub.4Cl (35 mg, 0.65 mmol)
and NaN.sub.3 (43 mg, 0.65 mmol) at room temperature. The reaction
mixture was stirred for 4 h at 110.degree. C. in the microwave, at
which stage 50% of the starting material was converted to the
desired product. The reaction mixture was purified by preparative
HPLC to provide
N-(2-tert-butyl-1-(2-hydroxy-3-(2H-tetrazol-5-yl)propyl-1H-indol-5-yl)-1--
(2,2-difluorobenzo-[d][1,3]dioxol-5-yl)cyclopropanecarboxamide. MS
(ESI) m/e (M+H.sup.+) 539.0.
Example 109
4-(2-tert-Butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclo-propaneca-
rboxamido)-1H-indol-1-yl)-3-hydroxybutanoic acid
##STR00775##
[0661] A solution of
N-(2-tert-butyl-1-(3-cyano-2-hydroxypropyl)-1H-indol-5-yl)-1-(2,2-difluor-
obenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide (14 mg, 0.028
mmol) in methanol (0.8 mL) and 4 M NaOH (0.8 mL) was stirred at
60.degree. C. for 4 h. The reaction mixture was neutralized with 4
M HCl and concentrated. The residue was purified by preparative
HPLC to provide
4-(2-tert-butyl-5-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropaneca-
rboxamido)-1H-indol-1-yl)-3-hydroxybutanoic acid. MS (ESI) m/e
(M+H.sup.+) 515.0.
Example 110
N-(1-(2-(2H-Tetrazol-5-yl)ethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][1-
,3]dioxol-5-yl)cyclopropanecarboxamide
##STR00776##
[0662]
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(2-cyanoethyl)indoli-
n-5-yl)-cyclopropanecarboxamide
[0663] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(2-chloroethyl)indolin-5-y-
l)cyclopropanecarboxamide (66 mg, 0.15 mmol) in ethanol (0.8 mL)
and water (0.4 mL) were added NaCN (22 mg, 0.45 mmol) and KI (cat)
at room temperature. The reaction mixture was stirred for 30 min at
110.degree. C. in the microwave before being purified by column
chromatography on silica gel (5-15% ethyl acetate/hexanes) to
provide
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(2-cyano-ethyl)indolin-5-y-
l)cyclopropanecarboxamide (50 mg, 77%).
##STR00777##
N-(1-(2-(2H-Tetrazol-5-yl)ethyl)-2-tert-butylbutyl-1H-indol-5-yl)-1-(benz-
o[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[0664] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1-(2-cyano-ethyl)indolin-5-y-
l)cyclopropanecarboxamide (50 mg, 0.12 mmol) in anhydrous DMF (2.6
mL) was added NH.sub.4CL (230 mg, 4.3 mmol) and NaN.sub.3 (280 mg,
4.3 mmol). The reaction mixture was stirred for 30 min at
110.degree. C. in the microwave, filtrated, and purified by
preparative HPLC. The solid residue was dissolved in CDCl.sub.3 (3
mL) and briefly (2 to 4 min) exposed to daylight, which initiated a
color change (purple). After 2 h of stirring open to the atmosphere
at room temperature, the solvent was removed and the residue was
purified by preparative HPLC to give
N-(1-(2-(2H-tetrazol-5-yl)ethyl)-2-tert-butyl-1H-indol-5-yl)-1-(benzo[d][-
1,3]dioxol-5-yl)cyclopropanecarboxamide. MS (ESI) m/e (M+H.sup.+)
473.0.
Example 111
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoro-1-((tetrahydro-2H-py-
ran-3-yl)methyl)-1H-indol-5-yl)cyclopropanecarboxamide
##STR00778##
[0666] To a solution of
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoroindolin-5-yl)cyclopr-
opane-carboxamide (150 mg, 0.38 mmol) in anhydrous dichloromethane
(2.3 mL) at room temperature under N.sub.2 was added
tetrahydropyran-3-carbaldehyde (54 mg, 0.47 mmol). After 20 min of
stirring, NaBH(OAc).sub.3 (110 mg, 0.51 mmol) was added in one
portion at room temperature. The reaction mixture was stirred for 6
h at room temperature before being purified by column
chromatography on silica gel (5-20% ethyl acetate/hexanes) to
provide
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoro-1-((tetrahydro-2H-p-
yran-3-yl)methyl)indolin-5-yl)cyclopropanecarboxamide (95 mg, 50%).
CDCl.sub.3 was added to the indoline and the solution was allowed
to stir overnight at ambient temperature. The solution was
concentrated to give
1-(benzo[d[1,3]dioxol-5-yl)-N-(2-tert-butyl-6-fluoro-1-((tetrahydro-2H-py-
ran-3-yl)methyl)-1H-indol-5-yl)cyclopropanecarboxamide. MS (ESI)
r/e (M+H+) 493.0.
Example 112
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-(2-hydroxypropan-2-yl)-1H-indol-5-yl)cyc-
lopropanecarboxamide
##STR00779##
[0668] Methyl
5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropane-carboxamido)-1H-indole-2-carb-
oxylate (100 mg, 0.255 mmol) was dissolved in anhydrous
tetrahydrofuran (2 mL) under an argon atmosphere. The solution was
cooled to 0.degree. C. in an ice water bath before methyllithium
(0.85 mL, 1.6 M in diethyl ether) was added by syringe. The mixture
was allowed to warm to room temperature. The crude product was then
partitioned between a saturated aqueous solution of sodium chloride
(5 mL) and dichloromethane (5 mL). The organic layers were
combined, dried over sodium sulfate, filtered, evaporated to
dryness, and purified on 12 g of silica gel utilizing a gradient of
20-80% ethyl acetate in hexanes to yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-(2-hydroxypropan-2-yl)-1H-indol-5-yl)cy-
clopropanecarboxamide (35 mg, 36%) as a white solid. ESI-MS m/z
calc. 378.2, found 379.1 (M+1).sup.+. Retention time of 2.18
minutes. .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.78 (s, 1H), 8.39
(s, 1H), 7.57 (d, J=1.7 Hz, 1H), 7.17 (d, J=8.6 Hz, 1H), 7.03-6.90
(m, 4H), 6.12 (d, J=1.5 Hz, 1H), 6.03 (s, 2H), 5.18 (s, 1H), 1.50
(s, 6H), 1.41-1.38 (m, 2H), 1.05-0.97 (m, 2H).
Example 113
N-(2-(1-Amino-2-methylpropan-2-yl)-1H-indol-5-yl)-1-(benzo[d][1,3]-dioxol--
5-yl)cyclopropanecarboxamide
##STR00780##
[0670] Trifluoroacetic acid (0.75 mL) was added to a solution of
tert-butyl
2-(5-(1-(benzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-1H-indol-2-yl)-
-2-methylpropylcarbamate (77 mg, 0.16 mmol) in dichloromethane (3
mL) and the mixture was stirred at room temperature for 1.5 h. The
mixture was evaporated, dissolved in dichloromethane, washed with
saturated sodium bicarbonate solution, dried over magnesium sulfate
and evaporated to dryness to give
N-(2-(1-amino-2-methylpropan-2-yl)-1H-indol-5-yl)-1-(benzo[d][1,3]dioxol--
5-yl)cyclopropanecarboxamide (53 mg, 86%). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 9.58 (s, 1H), 7.60 (d, J=1.6 Hz, 1H), 7.18-7.15
(m, 2H), 7.02-6.94 (m, 3H), 6.85 (d, J=7.8 Hz, 1H), 6.14 (d, J=1.2
Hz, 1H), 6.02 (s, 2H), 2.84 (s, 2H), 1.68 (dd, J=3.6, 6.7 Hz, 2H),
1.32 (s, 6H), 1.08 (dd, J=3.7, 6.8 Hz, 2H).
Example 114
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-(1-(dimethylamino)-2-methylpropan-2-yl)--
1H-indol-5-yl)cyclopropanecarboxamide
##STR00781##
[0672] To a solution of
N-(2-(1-amino-2-methylpropan-2-yl)-1H-indol-5-yl)-1-(benzo[d][1,3]dioxol--
5-yl)cyclopropanecarboxamide (20 mg, 0.051 mmol) in DMF (1 mL) was
added potassium carbonate (35 mg, 0.26 mmol) and iodomethane (7.0
.mu.L, 0.11 mmol). The mixture was stirred for 2 h. Water was added
and the mixture was extracted with dichloromethane. Combined
organic phases were dried over magnesium sulfate, evaporated,
coevaporated with toluene (3.times.) and purified by silica gel
chromatography (0-30% EtOAc in hexane) to give
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-(1-(dimethylamino)-2-methylpropan-2-yl)-
-1H-indol-5-yl)cyclopropanecarboxamide (7 mg, 33%). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 9.74 (s, 1H), 7.58 (d, J m 1.9 Hz,
1H), 7.20 (d, J=8.6 Hz, 1H), 7.15 (s, 1H), 7.01-6.95 (in, 31), 6.85
(d, J=7.9 Hz, 1H), 6.10 (d, J=0.9 Hz, 1H), 6.02 (s, 2H), 2.43 (s,
2H), 2.24 (s, 6H), 1.68 (dd, J=3.7, 6.7 Hz, 2H), 1.33 (s, 6H), 1.08
(dd, J=3.7, 6.8 Hz, 2H).
Example 115
N-(2-(1-Acetamido-2-methylpropan-2-yl)-1H-indol-5-yl)-1-(benzo[d][1,3]-dio-
xol-5-yl)cyclopropanecarboxamide
##STR00782##
[0674] To a solution of
N-(2-(1-amino-2-methylpropan-2-yl)-1H-indol-5-yl)-1-(benzo[d](1,3]dioxol--
5-yl)cyclopropanecarboxamide (21 mg, 0.054 mmol) in dichloromethane
(1 mL) was added pyridine (14 .mu.L, 0.16 mmol) followed by acetic
anhydride (6.0 .mu.L, 0.059 mmol). The mixture was stirred for 2 h.
Water was added and the mixture was extracted with dichloromethane,
evaporated, coevaporated with toluene (3.times.) and purified by
silica gel chromatography (60-100% ethylacetate in hexane) to give
N-(2-(l-acetamido-2-methylpropan-2-yl)-1H-indol-5-yl)-1-(benzo[d][1,3]-di-
oxol-5-yl)cyclopropanecarboxamide (17 mg, 73%). .sup.1H NMR (400
MHz, DMSO) .delta. 10.79 (s, 1H), 8.39 (s, 1H), 7.66 (t, J=6.2 Hz,
1H), 7.56 (d, J=1.7 Hz, 1H), 7.18-7.14 (m, 1H), 7.02-6.89 (m, 4H),
6.08 (d, J=1.5 Hz, 1H), 6.03 (s, 2H), 3.31 (d, J=6.2 Hz, 2H), 1.80
(s, 3H), 1.41-1.38 (m, 2H), 1.26 (s, 6H), 1.04-1.01 (m, 2H).
Example 116
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-(2-methyl-4-(1H-tetrazol-5-yl)butan-2-yl-
)-1H-indol-5-yl)cyclopropanecarboxamide
##STR00783##
[0676]
1-(Benzo[d[1,3]dioxol-5-yl)-N-(2-(4-cyano-2-methylbutan-2-yl)-1H-in-
dol-5-yl)cyclopropanecarboxamide (83 mg, 0.20 mmol) was dissolved
in N,N-dimethylformamide (1 mL) containing ammonium chloride (128
mg, 2.41 mmol), sodium azide (156 mg, 2.40 mmol), and a magnetic
stir bar. The reaction mixture was heated at 110.degree. C. for 40
minutes in a microwave reactor. The crude product was filtered and
then purified by preparative HPLC using a gradient of 0-99%
acetonitrile in water containing 0.05% trifluoroacetic acid to
yield
1-(benzo[d[1,3]dioxol-5-yl)-N-(2-(2-methyl-4-(1H-tetrazol-5-yl)butan-2-yl-
)-1H-indol-5-yl)cyclopropanecarboxamide. ESI-MS m/z calc. 458.2,
found 459.2 (M+1).sup.+. Retention time of 1.53 minutes. .sup.1H
NMR (400 MHz, CD.sub.3CN) 9.23 (s, 1H), 7.51-7.48 (m, 211), 7.19
(d, J=8.6 Hz, 1H), 7.06-7.03 (m, 2H), 6.95-6.89 (m, 2H), 6.17 (dd,
J=0.7, 2.2 Hz, 1H), 6.02 (s, 2H), 2.61-2.57 (m, 2H), 2.07-2.03 (m,
2H), 1.55-1.51 (m, 2H), 1.39 (s, 6H), 1.12-1.09 (m, 2H).
Example 117
1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-(piperidin-2-yl)-1H-indol-5-yl)cycloprop-
anecarboxamide
##STR00784##
[0678] tert-Butyl
2-(5-(1-(benzo[d[1,3]dioxol-5-yl)cyclo-propanecarboxamido)-1H-indol-2-yl)-
piperidine-1-carboxylate (55 mg, 0.11 mmol) was dissolved in
dichloromethane (2.5 mL) containing trifluoroacetic acid (1 mL).
The reaction mixture was stirred for 6 h at room temperature. The
crude product was purified by preparative HPLC using a gradient of
0-99% acetonitrile in water containing 0.05% trifluoroacetic acid
to yield
1-(benzo[d][1,3]dioxol-5-yl)-N-(2-(piperidin-2-yl)-1H-indol-5-yl)cyclopro-
panecarboxamide. ESI-MS m/z calc. 403.2, found 404.4 (M+1).sup.+.
Retention time of 0.95 minutes.
Example 118
5-tert-Butyl-1H-indol-6ylamine
##STR00785##
[0679] 2-Bromo-4-tert-butyl-phenylamine
[0680] To a solution of 4-tert-Butyl-phenylamine (447 g, 3.00 mol)
in DMF (500 mL) was added dropwise NBS (531 g, 3.00 mol) in DMF
(500 mL) at room temperature. Upon completion, the reaction mixture
was diluted with water and extracted with EtOAc. The organic layer
was washed with water, brine, dried over Na.sub.2SO.sub.4 and
concentrated. The crude product was directly used in the next step
without further purification.
##STR00786##
2-Bromo-4-tert-butyl-5-nitro-phenylamine
[0681] 2-Bromo-4-tert-butyl-phenylamine (160 g, 0.71 mol) was added
dropwise to H.sub.2SO.sub.4 (410 mL) at room temperature to yield a
clear solution. This clear solution was then cooled down to -5 to
-10.degree. C. A solution of KNO.sub.3 (83 g, 0.82 mol) in
H.sub.2SO.sub.4 (410 mL) was added dropwise while the temperature
was maintained between -5 to -10.degree. C. Upon completion, the
reaction mixture was poured into ice/water and extracted with
EtOAc. The combined organic layers were washed with 5%
Na.sub.2CO.sub.3 and brine, dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by a column chromatography
(ethyl acetate/petroleum ether 1:10) to give
2-bromo-4-tert-butyl-5-nitro-phenylamine as a yellow solid (150 g,
78%).
##STR00787##
4-tert-Butyl-5-nitro-2-trimethylsilanylethynyl-phenylamine
[0682] To a mixture of 2-bromo-4-tert-butyl-5-nitro-phenylamine
(27.3 g, 100 mmol) in toluene (200 mL) and water (100 mL) was added
Et.sub.3N (27.9 mL, 200 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (2.11 g,
3.00 mmol), CuI (950 mg, 0.500 mmol) and trimethylsilyl acetylene
(21.2 mL, 150 mmol) under a nitrogen atmosphere. The reaction
mixture was heated at 70.degree. C. in a sealed pressure flask for
2.5 h, cooled down to room temperature and filtered through a short
plug of Celite. The filter cake was washed with EtOAc. The combined
filtrate was washed with 5% NH.sub.4OH solution and water, dried
over Na.sub.2SO.sub.4 and concentrated. The crude product was
purified by column chromatography (0-10% ethyl acetate/petroleum
ether) to provide
4-tert-butyl-5-nitro-2-trimethylsilanylethynyl-phenylamine as a
brown viscous liquid (25 g, 81%).
##STR00788##
5-tert-Butyl-6-nitro-1H-indole
[0683] To a solution of
4-tert-butyl-5-nitro-2-trimethylsilanylethynyl-phenylamine (25 g,
86 mmol) in DMF (100 mL) was added CuI (8.2 g, 43 mmol) under a
nitrogen atmosphere. The mixture was heated at 135.degree. C. in a
sealed pressure flask overnight, cooled down to room temperature
and filtered through a short plug of Celite. The filter cake was
washed with EtOAc. The combined filtrate was washed with water,
dried over Na.sub.2SO.sub.4 and concentrated. The crude product was
purified by column chromatography (10-20% ethyl acetate/hexane) to
provide 5-tert-butyl-6-nitro-1H-indole as a yellow solid (13 g,
69%).
##STR00789##
5-tert-Butyl-1H-indol-6-ylamine
[0684] Raney Nickel (3 g) was added to
5-tert-butyl-6-nitro-1H-indole (15 g, 67 mmol) in methanol (100
mL). The mixture was stirred under hydrogen (1 atm) at 30.degree.
C. for 3 h. The catalyst was filtered off. The filtrate was dried
over Na.sub.2SO.sub.4 and concentrated. The crude dark brown
viscous oil was purified by column chromatography (10-20% ethyl
acetate/petroleum ether) to give 5-tert-butyl-1H-indol-6-ylamine as
a gray solid (11 g, 87%). .sup.1H NMR (300 MHz, DMSO-d6) .delta.
10.3 (br s, 1H), 7.2 (s, 1H), 6.9 (m, 1H), 6.6 (s, 1H), 6.1 (m,
1H), 4.4 (br s, 2H), 1.3 (s, 9H).
[0685] A person skilled in the chemical arts can use the examples
and schemes along with known synthetic methodologies to synthesize
compounds of the present invention, including the compounds in
Table 3, below.
TABLE-US-00003 TABLE 3 Physical data of exemplary compounds. Com-
pound LC/MS LC/RT No. M + 1 Min NMR 1 373.3 2.49 2 469.4 3.99 3
381.3 3.69 4 448.3 1.75 5 389.3 3.3 6 463 1.87 7 363.3 3.7 8 405.5
3.87 9 487.3 2.12 H NMR (400 MHz, DMSO-d6) 8.65 (s, 1H), 7.55 (d, J
= 1.7 Hz, 1H), 7.49 (d, J = 1.4 Hz, 1H), 7.38 (d, J = 8.3 Hz, 1H),
7.30-7.25 (m, 2H), 7.08 (dd, J = 8.8, 1.9 Hz, 1H), 6.11 (s, 1H),
4.31 (t, J = 7.4 Hz, 2H), 3.64 (t, J = 7.3 Hz, 2H), 3.20 (t, J =
7.6 Hz, 2H), 1.92 (t, J = 7.6 Hz, 2H), 1.45 (m, 2H), 1.39 (s, 6H),
1.10 (m, 2H) 10 388 3.34 11 452.3 2.51 12 527 2.36 13 498 1.85 14
404.5 1.18 15 369.2 3.81 16 419.2 2.24 17 389.2 2.02 H NMR (400
MHz, DMSO) 8.41 (s, 1H), 7.59 (d, J = 1.8 Hz, 1H), 7.15 (d, J = 8.6
Hz, 1H), 7.06-7.02 (m, 2H), 6.96-6.90 (m, 2H), 6.03 (s, 2H), 5.98
(d, J = 0.7 Hz, 1H), 4.06 (t, J = 6.8 Hz, 2H), 2.35 (t, J = 6.8 Hz,
2H), 1.42-1.38 (m, 2H), 1.34 (s, 6H), 1.05-1.01 (m, 2H) 18 395.3
3.6 H NMR (400 MHz, DMSO) 10.91 (s, 1H), 7.99 (s, 1H), 7.67 (d, J =
7.7 Hz, 1H), 7.08-6.92 (m, 4H), 6.09-6.03 (m, 3H), 1.47-1.42 (m,
2H), 1.31 (d, J = 7.3 Hz, 9H), 1.09-1.05 (m, 2H) 19 457.2 1.97 H
NMR (400 MHz, CD3CN) 7.50 (d, J = 1.9 Hz, 1H), 7.41 (d, J = 1.6 Hz,
2H), 7.36 (dd, J = 1.7, 8.3 Hz, 1H), 7.29-7.24 (m, 2H), 7.02 (dd, J
= 2.1, 8.8 Hz, 1H), 6.24 (s, 1H), 4.40 (t, J = 7.1 Hz, 2H), 3.80
(t, J = 7.1 Hz, 2H), 1.59-1.55 (m, 2H), 1.50 (s, 9H), 1.15-1.12 (m,
2H) 20 375.5 3.71 21 496 206 22 421.14 1.53 23 363.3 3.62 24 378.5
2.66 25 417.5 3.53 26 454.3 3.18 27 596.2 2.58 28 379.3 2.92 29 481
1.69 30 504.2 1.95 31 517 1.92 32 403.5 3.5 H NMR (400 MHz, DMSO)
10.76 (s, 1H), 8.72 (s, 1H), 7.79 (d, J = 2.3 Hz, 1H), 7.62 (dd, J
= 2.4, 8.6 Hz, 1H), 7.55 (d, J = 1.5 Hz, 1H), 7.14 (d, J = 8.6 Hz,
1H), 7.05-7.01 (m, 2H), 6.03 (d, J = 1.6 Hz, 1H), 4.54 (t, J = 6.4
Hz, 2H), 2.79 (t, J = 6.4 Hz, 2H), 1.44 (m, 2H), 1.32 (s, 9H), 1.03
(m, 2H) 33 321.3 2.98 34 450.2 2.02 35 395.1 3.59 36 509 2.01 37
447.2 2.02 38 379.1 2.16 H NMR (400 MHz, DMSO) 10.78 (s, 1H), 8.39
(s, 1H), 7.57 (d, J = 1.7 Hz, 1H), 7.17 (d, J = 8.6 Hz, 1H),
7.03-6.90 (m, 4H), 6.12 (d, J = 1.5 Hz, 1H), 6.03 (s, 2H), 5.18 (s,
1H), 1.50 (s, 6H), 1.41-1.38 (m, 2H), 1.05-0.97 (m, 2H) 39 373.3
3.74 40 372.8 3.8 41 397.3 3.41 H NMR (400 MHz, DMSO) 11.44 (s,
1H), 8.52 (s, 1H), 7.85 (d, J = 1.2 Hz, 2H), 7.71 (d, J = 1.7 Hz,
1H), 7.47-7.43 (m, 2H), 7.32-7.26 (m, 2H), 7.12 (dd, J = 2.0, 8.7
Hz, 1H), 7.04 (d, J = 1.6 Hz, 1H), 6.97-6.90 (m, 2H), 6.84 (d, J =
1.3 Hz, 1H), 6.03 (s, 2H), 1.43-1.40 (m, 2H), 1.07-1.03 (m, 2H) 42
505.3 2.23 H NMR (400 MHz, DMSO-d6) 8.33 (s, 1H), 7.52 (s, 1H),
7.42-7.39 (m, 2H), 7.33-7.25 (m, 2H), 6.14 (s, 1H), 4.99 (s, 1H),
4.31-4.27 (m, 3H), 3.64 (t, J = 7.0 Hz, 2H), 3.20 (t, J = 7.6 Hz,
2H), 1.91 (t, J = 7.6 Hz, 2H), 1.46 (m, 2H), 1.39 (s, 6H), 1.13 (m,
2H) 43 505.4 1.97 44 407.7 1.76 H NMR (400 MHz, DMSO) 10.31 (s,
1H), 8.34 (s, 1H), 7.53 (d, J = 1.8 Hz, 1H), 7.03 (d, J = 1.6 Hz,
1H), 6.97-6.90 (m, 3H), 6.05-6.03 (m, 3H), 4.72 (s, 2H), 1.40-1.38
(m, 2H), 1.34 (s, 9H), 1.04-1.00(m, 2H) 45 497.2 2.26 46 391.3 3.41
47 377.5 3.48 48 427.5 4.09 49 402.2 3.06 50 421.1 1.81 51 407.5
3.34 52 464.3 2.87 53 405.3 3.65 54 375 1.84 55 505.4 1.96 56 335.3
3.18 57 445.2 3.27 58 491 1.88 59 478 1.98 60 413.3 3.95 61 402.5
3.71 62 393.3 1.98 63 407.2 2.91 64 505.4 1.98 65 377.5 3.53 66
417.5 4.06 67 333.3 3.53 68 397.3 3.86 69 506 1.67 70 501 2.1 71
335.3 3.22 72 487 1.93 73 417.5 3.88 74 395 1.95 75 548 1.64 76
418.3 2.9 77 377.3 3.87 78 363.3 3.48 79 476 1.8 80 447.3 2.18 81
492.4 2 82 564.3 1.35 83 467.3 1.72 84 445.2 3.08 85 389.5 3.86 86
374.3 3.11 87 435 3.87 88 465 1.89 89 411.3 3.89 90 449.3 3.92 91
393.3 3.12 92 469.6 1.75 93 476.5 2.88 94 377.5 3.41 95 375.3 3.43
H NMR (400 MHz, DMSO) 10.52 (s, 1H), 8.39 (s, 1H), 7.46 (d, J = 1.8
Hz, 1H), 7.10-6.89 (m, 5H), 6.03 (s, 2H), 2.68-2.65 (m, 2H),
2.56-2.54 (m, 2H), 1.82-1.77 (m, 4H), 1.41-1.34 (m, 2H), 1.04-0.97
(m, 2H) 96 346.1 3.1 97 367.3 3.72 98 440.3 3.26 99 393.1 3.18 H
NMR (400 MHz, DMSO-d6) 11.80 (s, 1H), 8.64 (s, 1H), 7.83 (m, 1H),
7.33-7.26 (m, 2H), 7.07 (m, 1H), 7.02 (m, 1H), 6.96-6.89 (m, 2H),
6.02 (s, 2H), 4.33 (q, J = 7.1 Hz, 2H), 1.42-1.39 (m, 2H), 1.33 (t,
J = 7.1 Hz, 3H), 1.06-1.03 (m, 2H) 100 421.3 1.85 H NMR (400 MHz,
DMSO) 13.05 (s, 1H), 9.96 (d, J = 1.6 Hz, 1H), 7.89 (d, J = 1.9 Hz,
1H), 7.74 (d, J = 2.0 Hz, 1H), 7.02 (d, J = 1.6 Hz, 1H), 6.96-6.88
(m, 2H), 6.22 (d, J = 2.3 Hz, 1H), 6.02 (s, 2H), 1.43-1.40 (m, 2H),
1.37 (s, 9H), 1.06-1.02 (m, 2H) 101 387.5 2.51 102 479 3.95 103
420.3 3.12 104 469.5 3.97 105 391.3 2.04 106 375.2 2.82 107 349.3
3.33 108 503.3 1.88 109 451.5 1.59 110 361.5 3.7 111 391.3 3.65 112
335.3 3.03 113 496.5 1.68 114 381.5 3.72 115 390.3 3.22 116 397.3
3.52 H NMR (400 MHz, DMSO-d6) 11.27 (d, J = 1.9 Hz, 1H), 8.66 (s,
1H), 8.08 (d, J = 1.6 Hz, 1H), 7.65-7.61 (m, 3H), 7.46-7.40 (m,
2H), 7.31 (d, J = 8.7 Hz, 1H), 7.25-7.17 (m, 2H), 7.03 (d, J = 1.6
Hz, 1H), 6.98-6.87 (m, 2H), 6.02 (s, 2H), 1.43-1.39 (m, 2H),
1.06-1.02 (m, 2H) 117 377.5 3.77 118 515.3 2.3 119 381.3 3.8 120
464.2 2.1 121 465 1.74 122 395.2 3.74 123 383.3 3.52 124 388.5 3.56
125 411.3 3.85 126 459.2 1.53 H NMR (400 MHz, CD3CN) 9.23 (s, 1H),
7.51-7.48 (m, 2H), 7.19 (d, J = 8.6 Hz, 1H), 7.06-7.03 (m, 2H),
6.95-6.89 (m, 2H), 6.17 (dd, J = 0.7, 2.2 Hz, 1H), 6.02 (s, 2H),
2.61- 2.57 (m, 2H), 2.07-2.03 (m, 2H), 1.55-1.51 (m, 2H), 1.39 (s,
6H), 1.12-1.09 (m, 2H) 127 408.5 2.48 128 393 3.26 129 420.2 2.16
130 406.3 2.88 131 473.3 4.22 132 417.3 3.8 133 465 1.74 134 464.3
2.91 135 347.3 3.42 136 511 2.35 137 455.5 3.29 138 393.3 3.54 139
335.1 3.08 140 434.5 2.74 141 381.3 2.91 142 431.5 3.97 143 539
1.89 144 515 1.89 145 407.5 3.6 146 379.5 1.51 147 409.3 4 148
392.2 1.22 149 375.3 3.37 150 377.3 3.61
151 377.22 3.96 152 504.5 1.99 153 393.1 3.47 154 363.3 3.52 155
321.3 3.13 156 407.5 3.2 157 406.3 1.43 158 379.3 1.89 159 451 3.34
160 375.3 3.82 161 355.1 3.32 162 475 2.06 163 437.2 2.35 164 379.2
2.76 165 462 3.44 166 465.2 2.15 167 455.2 2.45 168 451 1.65 169
528 1.71 170 374.3 3.4 171 449.5 1.95 172 381.3 3.8 173 346.3 2.93
174 483.1 2.25 175 411.2 3.85 176 431.5 4.02 177 485.5 4.02 178
528.5 1.18 179 473 1.79 180 479 2.15 181 387.5 2.56 182 365.3 3.13
183 493 2.3 184 461.3 2.4 H NMR (400 MHz, DMSO-d6) 10.89 (s, 1H),
8.29 (s, 1H), 7.52 (s, 1H), 7.42-7.37 (m, 2H), 7.32 (dd, J = 8.3,
1.4 Hz, 1H), 7.01 (d, J = 10.9 Hz, 1H), 6.05 (d, J = 1.7 Hz, 1H),
4.29 (t, J = 5.0 Hz, 1H), 3.23 (m, 2H), 1.81 (t, J = 7.7 Hz, 2H),
1.46 (m, 2H), 1.29 (s, 6H), 1.13 (m, 2H) 185 377.5 3.63 186 464
1.46 187 339.1 3.2 188 435.5 1.64 189 392.3 2.18 190 435.5 3.67 H
NMR (400 MHz, DMSO) 11.83 (s, 1H), 10.76 (s, 1H), 8.53 (s, 1H),
7.93 (d, J = 1.8 Hz, 1H), 7.60 (dd, J = 2.3, 8.5 Hz, 1H), 7.53 (d,
J = 1.4 Hz, 1H), 7.14 (d, J = 8.6 Hz, 1H), 7.02-6.97 (m, 2H), 6.02
(d, J = 1.5 Hz, 1H), 3.71 (t, J = 6.2 Hz, 2H), 3.37 (t, J = 6.2 Hz,
2H), 3.25 (s, 3H), 1.44 (m, 2H), 1.32 (s, 9H), 1.08 (m, 2H) 191
421.3 3.32 192 404.4 0.95 193 451 1.71 194 465 1.69 195 434.2 2.29
196 363.3 3.4 197 501 1.91 198 411.2 3.14 199 439 1.89 200 434.4
1.53 201 462 3.22 202 351.3 2.59 203 495.2 2.71 204 435 3.94 205
397.3 3.69 206 493 2.26 207 487 1.87 208 391.3 2.94 209 397.2 3.3
210 487.2 1.85 H NMR (400 MHz, CD3CN) 7.50 (d, J = 2.0 Hz, 1H),
7.41 (d, J = 1.6 Hz, 2H), 7.37-7.32 (m, 2H), 7.25 (d, J = 8.3 Hz,
1H), 6.98 (dd, J = 2.1, 8.8 Hz, 1H), 6.27 (d, J = 0.6 Hz, 1H),
4.40-4.28 (m, 2H), 4.12-4.06 (m, 1H), 3.59-3.51 (m, 2H), 1.59-1.50
(m, 2H), 1.47 (s, 9H), 1.15-1.12 (m, 2H) 211 381.3 3.69 212 461
2.04 213 469 1.72 214 363.3 3.48 215 432.3 3.07 216 403.5 3.94 217
420.4 1.27 218 475 2.2 219 484.3 1.84 220 419.3 3.87 221 486.3 0.91
222 391.3 3.01 223 398.3 1.3 224 349.2 2.54 225 375.5 3.74 226
377.5 3.47 H NMR (400 MHz, DMSO-d6) 10.76 (s, 1H), 8.39 (s, 1H),
7.55 (s, 1H), 7.15-7.13 (m, 1H), 7.03-6.89 (m, 4H), 6.03 (m, 3H),
1.41-1.38 (m, 2H), 1.32 (s, 9H), 1.04-1.01 (m, 2H) 227 393.3 2.03
228 398.3 1.24 229 487.2 1.78 230 361.1 3.47 231 435.5 2.12 232
321.3 2.91 233 413.3 3.77 234 393.3 1.58 235 465 1.92 236 361.3
3.18 237 421 1.8 238 405.5 3.79 239 544.3 1.4 240 405.3 3.9 241 462
1.74 242 550 1.68 243 395.2 1.98 244 517.3 1.94 245 372.2 3.59 246
361.3 3.58 247 490 1.95 248 407.3 1.52 H NMR (400 MHz, DMSO) 10.74
(d, J = 1.2 Hz, 1H), 8.40 (s, 1H), 7.54 (d, J = 1.8 Hz, 1H), 7.15
(d, J = 8.6 Hz, 1H), 7.03-6.90 (m, 4H), 6.03-6.00 (m, 3H),
3.26-3.22 (m, 2H), 1.85-1.80 (m, 2H), 1.41-1.38 (m, 2H), 1.31 (s,
6H), 1.05-1.01 (m, 2H) 249 393.3 3.32 250 406.2 2.08 251 511 2.39
252 379.3 3.3 253 383 3.46 254 401.2 3.26 255 398.3 1.38 256 512.5
1.96 257 389.2 3.05 258 321.3 3.02 259 392.1 2.74 260 462 1.81 261
453 1.91 262 349.3 3.22 263 391.1 3.67 H NMR (400 MHz, DMSO)
1.01-1.05 (dd, J = 4.0, 6.7 Hz, 2H), 1.41-1.39 (m, 11H), 3.81 (s,
3H), 6.03 (s, 2H), 6.15 (s, 1H), 6.96-6.90 (m, 2H), 7.02 (d, J =
1.6 Hz, 1H), 7.09 (dd, J = 2.0, 8.8 Hz, 1H), 7.25 (d, J = 8.8 Hz,
1H), 7.60 (d, J = 1.9 Hz, 1H), 8.46 (s, 1H) 264 421.3 1.66 H NMR
(400 MHz, CD3CN) 8.78 (s, 1H), 7.40 (m, 1H), 7.33 (s, 1H), 7.08 (m,
1H), 6.95-6.87 (m, 3H), 6.79 (m, 1H), 5.91 (s, 2H), 3.51 (dd, J =
5.9, 7.8 Hz, 2H), 2.92-2.88 (m, 2H), 2.64 (t, J = 5.8 Hz, 1H), 1.50
(m, 2H), 1.41 (s, 9H), 1.06 (m, 2H) 265 475 2.15 266 347.3 3.32 267
420.5 1.81 268 416.2 1.76 269 485 2.06 270 395.3 3.89 271 492 1.59
272 405.5 3.96 273 547.2 1.65 274 631.6 1.91 275 590.4 2.02 276
465.7 1.79 277 411.3 2.14 278 385.3 1.99 279 425.3 2.19 280 473.2
1.74 281 469.4 2.02 H NMR (400 MHz, DMSO) 8.82 (s, 1H), 7.84 (d, J
= 1.7 Hz, 1H), 7.55-7.51 (m, 2H), 7.40-7.35 (m, 2H), 7.29 (dd, J =
1.7, 8.3 Hz, 1H), 7.04 (s, 1H), 4.98 (t, J = 5.6 Hz, 1H), 4.27 (t,
J = 6.1 Hz, 2H), 3.67 (q, J = 6.0 Hz, 2H), 1.48 (dd, J = 4.0, 6.7
Hz, 2H), 1.13 (dd, J = 4.1, 6.8 Hz, 2H) 282 644.4 1.83 283 544.6
1.97 284 465.4 1.56 285 485.2 1.8 286 475.2 1.87 287 564.2 1.95 288
512.5 1.89 H NMR (400 MHz, DMSO) 8.77 (s, 1H), 7.97 (s, 1H), 7.51
(s, 1H), 7.43-7.40 (m, 2H), 7.33 (d, J = 8.2 Hz, 1H), 6.36 (s, 1H),
4.99-4.97 (m, 2H), 4.52 (d, J = 13.1 Hz, 1H), 4.21 (dd, J = 9.2,
15.2 Hz, 1H), 3.86 (m, 1H), 3.51-3.36 (m, 2H), 1.51-1.48 (m, 2H),
1.43 (s, 9H), 1.17-1.15 (m, 2H) 289 437.3 1.6 290 499.5 1.81 H NMR
(400 MHz, DMSO) 8.82 (s, 1H), 7.83 (d, J = 1.7 Hz, 1H), 7.55-7.50
(m, 2H), 7.39-7.28 (m, 3H), 7.03 (s, 1H), 4.97 (d, J = 5.6 Hz, 1H),
4.83 (t, J = 5.6 Hz, 1H), 4.33 (dd, J = 3.4, 15.1 Hz, 1H), 4.09
(dd, J = 8.7, 15.1 Hz, 1H), 3.80-3.78 (m, 1H), 3.43-3.38 (m, 1H),
3.35-3.30 (m, 1H), 1.49-1.46 (m, 2H), 1.14-1.11 (m, 2H) 291 455.4
2.02 H NMR (400 MHz, DMSO) 8.62 (s, 1H), 7.56 (s, 1H), 7.50 (s,
1H), 7.38 (d, J = 8.3 Hz, 1H), 7.29 (dd, J = 1.5, 8.3 Hz, 1H), 7.23
(d, J = 8.7 Hz, 1H), 7.06 (dd, J = 1.7, 8.7 Hz, 1H), 6.19 (s, 1H),
4.86 (t, J = 5.4 Hz, 1H), 4.03 (t, J = 6.1 Hz, 2H), 3.73 (qn, J =
8.5 Hz, 1H), 3.57 (q, J = 5.9 Hz, 2H), 2.39-2.33 (m, 2H), 2.18-1.98
(m, 3H), 1.88-1.81 (m, 1H), 1.47-1.44 (m, 2H), 1.11-1.09 (m, 2H)
292 578.4 1.99 293 630.4 1.8 294 443.4 1.98 H NMR (400 MHz, DMSO)
8.62 (s, 1H), 7.55 (d, J = 1.8 Hz, 1H), 7.50 (d, J = 1.5 Hz, 1H),
7.38 (d, J = 8.3 Hz, 1H), 7.30-7.24 (m, 2H), 7.05 (dd, J = 2.0, 8.8
Hz, 1H), 6.13 (s, 1H), 4.88 (t, J = 5.5 Hz, 1H), 4.14 (t, J = 6.1
Hz, 2H), 3.61 (m, 2H), 3.21 (septet, J = 6.8 Hz, 1H), 1.47-1.44 (m,
2H), 1.26 (d, J = 6.8 Hz, 6H), 1.11-1.08 (m, 2H) 295 482.3 2 H NMR
(400 MHz, DMSO) 8.78 (s, 1H), 7.92 (s, 1H), 7.51 (s, 1H), 7.45 (s,
1H), 7.41 (d, J = 8.3 Hz, 1H), 7.33 (d, J = 8.4 Hz, 1H), 6.34 (s,
1H), 5.01 (t, J = 5.7 Hz, 1H), 4.41 (t, J = 6.6 Hz, 2H), 3.68 (m,
2H), 1.51-1.47 (m, 2H), 1.42 (s, 9H), 1.19-1.15 (m, 2H) 296 438.7
2.12 H NMR (400 MHz, DMSO) 11.43 (s, 1H), 8.74 (s, 1H), 7.63 (s,
1H), 7.51 (s, 1H), 7.45-7.40 (m, 2H), 7.33 (dd, J = 1.4, 8.3 Hz,
1H), 6.25 (d, J = 1.5 Hz, 1H), 1.51-1.48 (m, 2H), 1.34 (s, 9H),
1.17-1.14 (m, 2H)
297 449.3 1.6 298 517.5 1.64 299 391.5 2.05 300 449.3 1.59 301
501.2 1.93 302 503.5 1.63 303 437.3 1.6 304 425.1 2.04 H NMR (400
MHz, DMSO) 12.16 (s, 1H), 8.80 (s, 1H), 7.83 (s, 1H), 7.51 (d, J =
1.4 Hz, 1H), 7.39-7.28 (m, 4H), 6.95 (s, 1H), 1.48 (dd, J = 4.0,
6.6 Hz, 2H), 1.13 (dd, J = 4.0, 6.7 Hz, 2H) 305 459.2 1.67 306
558.4 2.05
VII. Assays for Detecting and Measuring .DELTA.F508-CFTR Correction
Properties of Compounds
[0686] Membrane Potential Optical Methods for Assaying
.DELTA.F508-CFTR Modulation Properties of Compounds
[0687] The optical membrane potential assay utilized
voltage-sensitive FRET sensors described by Gonzalez and Tsien
(See, Gonzalez, J. E. and R. Y. Tsien (1995) "Voltage sensing by
fluorescence resonance energy transfer in single cells" Biophys J
69(4): 1272-80, and Gonzalez, J. E. and R. Y. Tsien (1997)
"Improved indicators of cell membrane potential that use
fluorescence resonance energy transfer" Chem Biol 4(4): 269-77) in
combination with instrumentation for measuring fluorescence changes
such as the Voltage/Ion Probe Reader (VIPR) (See, Gonzalez, J. E.,
K. Oades, et al. (1999) "Cell-based assays and instrumentation for
screening ion-channel targets" Drug Discov Today 4(9):
431-439).
[0688] These voltage sensitive assays are based on the change in
fluorescence resonant energy transfer (FRET) between the
membrane-soluble, voltage-sensitive dye, DiSBAC.sub.2 (3), and a
fluorescent phospholipid, CC2-DMPE, which is attached to the outer
leaflet of the plasma membrane and acts as a FRET donor. Changes in
membrane potential (Vn) cause the negatively charged
DiSBAC.sub.2(3) to redistribute across the plasma membrane and the
amount of energy transfer from CC2-DMPE changes accordingly. The
changes in fluorescence emission were monitored using VIPR.TM. II,
which is an integrated liquid handler and fluorescent detector
designed to conduct cell-based screens in 96- or 384-well
microtiter plates.
[0689] Identification of Correction Compounds
[0690] To identify small molecules that correct the trafficking
defect associated with .DELTA.F508-CFTR; a single-addition HTS
assay format was developed. The cells were incubated in serum-free
medium for 16 hrs at 37.degree. C. in the presence or absence
(negative control) of test compound. As a positive control, cells
plated in 384-well plates were incubated for 16 hrs at 27.degree.
C. to "temperature-correct" .DELTA.F508-CFTR. The cells were
subsequently rinsed 3.times. with Krebs Ringers solution and loaded
with the voltage-sensitive dyes. To activate .DELTA.F508-CFTR, 10
.mu.M forskolin and the CFTR potentiator, genistein (20 .mu.M),
were added along with Cl.sup.--free medium to each well. The
addition of Cl.sup.--free medium promoted Cl.sup.- efflux in
response to .DELTA.F508-CFTR activation and the resulting membrane
depolarization was optically monitored using the FRET-based
voltage-sensor dyes.
[0691] Identification of Potentiator Compounds
[0692] To identify potentiators of .DELTA.F508-CFTR, a
double-addition HTS assay format was developed. During the first
addition, a Cl.sup.--free medium with or without test compound was
added to each well. After 22 sec, a second addition of
Cl.sup.--free medium containing 2-10 .mu.M forskolin was added to
activate .DELTA.F508-CFTR. The extracellular Cl.sup.- concentration
following both additions was 28 mM, which promoted Cl.sup.- efflux
in response to .DELTA.F508-CFTR activation and the resulting
membrane depolarization was optically monitored using the
FRET-based voltage-sensor dyes. SolutionsBath Solution #1: (in mM)
NaCl 160, KCl 4.5, CaCl.sub.2 2, MgCl.sub.2 1, HEPES 10, pH 7.4
with NaOH.
[0693] Chloride-free bath solution: Chloride salts in Bath Solution
#1 are substituted with gluconate salts.
[0694] CC2-DMPE: Prepared as a 10 mM stock solution in DMSO and
stored at -20.degree. C.
[0695] DiSBAC.sub.2(3): Prepared as a 10 mM stock in DMSO and
stored at -20.degree. C.
[0696] Cell Culture
[0697] NIH3T3 mouse fibroblasts stably expressing .DELTA.F508-CFTR
are used for optical measurements of membrane potential. The cells
are maintained at 37.degree. C. in 5% CO.sub.2 and 90% humidity in
Dulbecco's modified Eagle's medium supplemented with 2 mM
glutamine, 10% fetal bovine serum, 1.times.NEAA, .beta.-ME,
1.times. pen/strep, and 25 mM HEPES in 175 cm.sup.2 culture flasks.
For all optical assays, the cells were seeded at 30,000/well in
384-well matrigel-coated plates and cultured for 2 hrs at
37.degree. C. before culturing at 27.degree. C. for 24 hrs. for the
potentiator assay. For the correction assays, the cells are
cultured at 27.degree. C. or 37.degree. C. with and without
compounds for 16-24 hours Electrophysiological Assays for assaying
.DELTA.F508-CFTR modulation properties of compounds Ussing Chamber
AssayUssing chamber experiments were performed on polarized
epithelial cells expressing .DELTA.F508-CFTR to further
characterize the .DELTA.F508-CFTR modulators identified in the
optical assays. FRT.sup..DELTA.F508-CFTR epithelial cells grown on
Costar Snapwell cell culture inserts were mounted in an Ussing
chamber (Physiologic Instruments, Inc., San Diego, Calif.), and the
monolayers were continuously short-circuited using a Voltage-clamp
System (Department of Bioengineering, University of Iowa, Iowa,
and, Physiologic Instruments, Inc., San Diego, Calif.).
Transepithelial resistance was measured by applying a 2-mV pulse.
Under these conditions, the FRT epithelia demonstrated resistances
of 4 .OMEGA./cm.sup.2 or more. The solutions were maintained at
27.degree. C. and bubbled with air. The electrode offset potential
and fluid resistance were corrected using a cell-free insert. Under
these conditions, the current reflects the flow of CT through
.DELTA.F508-CFTR expressed in the apical membrane. The I.sub.SC was
digitally acquired using an MP100A-CE interface and AcqKnowledge
software (v3.2.6; BIOPAC Systems, Santa Barbara, Calif.).
[0698] Identification of Correction Compounds
[0699] Typical protocol utilized a basolateral to apical membrane
Cl.sup.- concentration gradient. To set up this gradient, normal
ringer was used on the basolateral membrane, whereas apical NaCl
was replaced by equimolar sodium gluconate (titrated to pH 7.4 with
NaOH) to give a large Cl.sup.- concentration gradient across the
epithelium. All experiments were performed with intact monolayers.
To fully activate .DELTA.F508-CFTR, forskolin (10 .mu.M) and the
PDE inhibitor, IBMX (100 .mu.M), were applied followed by the
addition of the CFTR potentiator, genistein (50 .mu.M).
[0700] As observed in other cell types, incubation at low
temperatures of FRT cells stably expressing .DELTA.F508-CFTR
increases the functional density of CFTR in the plasma membrane. To
determine the activity of correction compounds, the cells were
incubated with 10 .mu.M of the test compound for 24 hours at
37.degree. C. and were subsequently washed 3.times. prior to
recording. The cAMP- and genistein-mediated I.sub.SC in
compound-treated cells was normalized to the 27.degree. C. and
37.degree. C. controls and expressed as percentage activity.
Preincubation of the cells with the correction compound
significantly increased the cAMP- and genistein-mediated I.sub.SC
compared to the 37.degree. C. controls.
[0701] Identification of Potentiator Compounds
[0702] Typical protocol utilized a basolateral to apical membrane
Cl.sup.- concentration gradient. To set up this gradient, normal
ringers was used on the basolateral membrane and was permeabilized
with nystatin (360 .mu.g/ml), whereas apical NaCl was replaced by
equimolar sodium gluconate (titrated to pH 7.4 with NaOH) to give a
large Cl.sup.- concentration gradient across the epithelium. All
experiments were performed 30 min after nystatin permeabilization.
Forskolin (10 .mu.M) and all test compounds were added to both
sides of the cell culture inserts. The efficacy of the putative
.DELTA.F508-CFTR potentiators was compared to that of the known
potentiator, genistein.
[0703] Solutions
[0704] Basolateral solution (in mM): NaCl (135), CaCl.sub.2 (1.2),
MgCl.sub.2 (1.2), K.sub.2HPO.sub.4 (2.4), KHPO.sub.4 (0.6),
N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) (10),
and dextrose (10). The solution was titrated to pH 7.4 with
NaOH.
[0705] Apical solution (in mM): Same as basolateral solution with
NaCl replaced with Na Gluconate (135).
[0706] Cell Culture
[0707] Fisher rat epithelial (FRT) cells expressing
.DELTA.F508-CFTR (FRT.sup..DELTA.F508-CFTR) were used for Ussing
chamber experiments for the putative .DELTA.F508-CFTR modulators
identified from our optical assays. The cells were cultured on
Costar Snapwell cell culture inserts and cultured for five days at
37.degree. C. and 5% CO.sub.2 in Coon's modified Ham's F-12 medium
supplemented with 5% fetal calf serum, 100 U/ml penicillin, and 100
.mu.g/ml streptomycin. Prior to use for characterizing the
potentiator activity of compounds, the cells were incubated at
27.degree. C. for 16-48 hrs to correct for the .DELTA.F508-CFTR. To
determine the activity of corrections compounds, the cells were
incubated at 27.degree. C. or 37.degree. C. with and without the
compounds for 24 hours.
[0708] The macroscopic .DELTA.F508-CFTR current (I.sub..DELTA.F508)
in temperature- and test compound-corrected NIH3T3 cells stably
expressing .DELTA.F508-CFTR were monitored using the
perforated-patch, whole-cell recording. Briefly, voltage-clamp
recordings of I.sub..DELTA.F508 were performed at room temperature
using an Axopatch 200B patch-clamp amplifier (Axon Instruments
Inc., Foster City, Calif.). All recordings were acquired at a
sampling frequency of 10 kHz and low-pass filtered at 1 kHz.
Pipettes had a resistance of 5-6 M.OMEGA. when filled with the
intracellular solution. Under these recording conditions, the
calculated reversal potential for Cl.sup.- (Eca) at room
temperature was -28 mV. All recordings had a seal resistance >20
GO and a series resistance <15 M.OMEGA.. Pulse generation, data
acquisition, and analysis were performed using a PC equipped with a
Digidata 1320 A/D interface in conjunction with Clampex 8 (Axon
Instruments Inc.). The bath contained <250 .mu.l of saline and
was continuously perifused at a rate of 2 ml/min using a
gravity-driven perfusion system.
[0709] Identification of Correction Compounds
[0710] To determine the activity of correction compounds for
increasing the density of functional .DELTA.F508-CFTR in the plasma
membrane, we used the above-described perforated-patch-recording
techniques to measure the current density following 24-hr treatment
with the correction compounds. To fully activate .DELTA.F508-CFTR,
10 .mu.M forskolin and 20 .mu.M genistein were added to the cells.
Under our recording conditions, the current density following 24-hr
incubation at 27.degree. C. was higher than that observed following
24-hr incubation at 37.degree. C. These results are consistent with
the known effects of low-temperature incubation on the density of
.DELTA.F508-CFTR in the plasma membrane. To determine the effects
of correction compounds on CFTR current density, the cells were
incubated with 10 .mu.M of the test compound for 24 hours at
37.degree. C. and the current density was compared to the
27.degree. C. and 37.degree. C. controls (% activity). Prior to
recording, the cells were washed 3.times. with extracellular
recording medium to remove any remaining test compound.
Preincubation with 10 .mu.M of correction compounds significantly
increased the cAMP- and genistein-dependent current compared to the
37.degree. C. controls.
[0711] Identification of Potentiator Compounds
[0712] The ability of .DELTA.F508-CFTR potentiators to increase the
macroscopic F508-CFTR CI current (I.sub..DELTA.F508) in NIH3T3
cells stably expressing .DELTA.F508-CFTR was also investigated
using perforated-patch-recording techniques. The potentiators
identified from the optical assays evoked a dose-dependent increase
in I.sub..DELTA.F508 with similar potency and efficacy observed in
the optical assays. In all cells examined, the reversal potential
before and during potentiator application was around -30 mV, which
is the calculated E.sub.Cl (-28 mV).
[0713] Solutions
[0714] Intracellular solution (in mM): Cs-aspartate (90), CsCl
(50), MgCl.sub.2 (1), HEPES (10), and 240 .mu.g/ml amphotericin-B
(pH adjusted to 7.35 with CsOH).
[0715] Extracellular solution (in mM): N-methyl-D-glucamine
(NMDG)-Cl (150), MgCl.sub.2 (2), CaCl.sub.2 (2), HEPES (10) (pH
adjusted to 7.35 with HCl).
[0716] Cell Culture
[0717] NIH3T3 mouse fibroblasts stably expressing .DELTA.F508-CFTR
are used for whole-cell recordings. The cells are maintained at
37.degree. C. in 5% CO.sub.2 and 90% humidity in Dulbecco's
modified Eagle's medium supplemented with 2 mM glutamine, 10% fetal
bovine serum, 1.times.NEAA, .beta.-ME, 1.times. pen/strep, and 25
mM HEPES in 175 cm.sup.2 culture flasks. For whole-cell recordings,
2,500-5,000 cells were seeded on poly-L-lysine-coated glass
coverslips and cultured for 24-48 hrs at 27.degree. C. before use
to test the activity of potentiators; and incubated with or without
the correction compound at 37.degree. C. for measuring the activity
of correctors.
[0718] The single-channel activities of temperature-corrected
.DELTA.F508-CFTR stably expressed in NIH3T3 cells and activities of
potentiator compounds were observed using excised inside-out
membrane patch. Briefly, voltage-clamp recordings of single-channel
activity were performed at room temperature with an Axopatch 200B
patch-clamp amplifier (Axon Instruments Inc.). All recordings were
acquired at a sampling frequency of 10 kHz and low-pass filtered at
400 Hz. Patch pipettes were fabricated from Corning Kovar Sealing
#7052 glass (World Precision Instruments, Inc., Sarasota, Fla.) and
had a resistance of 5-8 MO when filled with the extracellular
solution. The 6F508-CFTR was activated after excision, by adding 1
mM Mg-ATP, and 75 nM of the cAMP-dependent protein kinase,
catalytic subunit (PKA; Promega Corp. Madison, Wis.). After channel
activity stabilized, the patch was perifused using a gravity-driven
microperfusion system. The inflow was placed adjacent to the patch,
resulting in complete solution exchange within 1-2 sec. To maintain
.DELTA.F508-CFTR activity during the rapid perifusion, the
nonspecific phosphatase inhibitor F (10 mM NaF) was added to the
bath solution. Under these recording conditions, channel activity
remained constant throughout the duration of the patch recording
(up to 60 min). Currents produced by positive charge moving from
the intra- to extracellular solutions (anions moving in the
opposite direction) are shown as positive currents. The pipette
potential (V.sub.p) was maintained at 80 mV.
[0719] Channel activity was analyzed from membrane patches
containing .ltoreq.2 active channels. The maximum number of
simultaneous openings determined the number of active channels
during the course of an experiment. To determine the single-channel
current amplitude, the data recorded from 120 sec of
.DELTA.F508-CFTR activity was filtered "off-line" at 100 Hz and
then used to construct all-point amplitude histograms that were
fitted with multigaussian functions using Bio-Patch Analysis
software (Bio-Logic Comp. France). The total microscopic current
and open probability (P.sub.o) were determined from 120 sec of
channel activity. The P.sub.o was determined using the Bio-Patch
software or from the relationship P.sub.o=I/i(N), where I=mean
current, i=single-channel current amplitude, and N=number of active
channels in patch.
[0720] Solutions
[0721] Extracellular solution (in mM): NMDG (150), aspartic acid
(150), CaCl.sub.2 (5), MgCl.sub.2 (2), and HEPES (10) (pH adjusted
to 7.35 with Tris base).
[0722] Intracellular solution (in mM): NMDG-CI (150), MgCl.sub.2
(2), EGTA (5), TES (10), and Tris base (14) (pH adjusted to 7.35
with HCl).
[0723] Cell Culture
[0724] NIH3T3 mouse fibroblasts stably expressing .DELTA.F508-CFTR
are used for excised-membrane patch-clamp recordings. The cells are
maintained at 37.degree. C. in 5% CO.sub.2 and 90% humidity in
Dulbecco's modified Eagle's medium supplemented with 2 mM
glutamine, 10% fetal bovine serum, 1.times.NEAA, .beta.-ME,
1.times. pen/strep, and 25 mM HEPES in 175 cm.sup.2 culture flasks.
For single channel recordings, 2,500-5,000 cells were seeded on
poly-L-lysine-coated glass coverslips and cultured for 24-48 hrs at
27.degree. C. before use.
[0725] Compounds of the invention are useful as modulators of ATP
binding cassette transporters. Using the procedures described
above, the activities, i.e., EC50s, of compounds of the present
invention have been measured to be from about 3.8 nM to about 13.5
.mu.M. Furthermore, using those methods described above, the
efficacies of compounds of the present invention have been measured
to be from about 35% to about 110%.
Other Embodiments
[0726] It is to be understood that while the invention has been
described in conjunction with the detailed description thereof; the
foregoing description is intended to illustrate and not limit the
scope of the invention, which is defined by the scope of the
appended claims. Other aspects, advantages, and modifications are
within the scope of the following claims.
* * * * *
References