U.S. patent application number 09/930734 was filed with the patent office on 2002-02-14 for 9h-pyrimido[4,5-b]indole derivatives: crf1 specific ligands.
This patent application is currently assigned to Neurogen Corporation. Invention is credited to Horvath, Raymond F.
Application Number | 20020019406 09/930734 |
Document ID | / |
Family ID | 22450710 |
Filed Date | 2002-02-14 |
United States Patent
Application |
20020019406 |
Kind Code |
A1 |
Horvath, Raymond F |
February 14, 2002 |
9H-pyrimido[4,5-b]indole derivatives: CRF1 specific ligands
Abstract
Disclosed are compounds of the formula: 1 wherein Ar, R.sup.1,
R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are defined herein, which
compounds are selective antagonists at CRF1 receptors and are
therefore useful in the diagnosis and treatment of stress related
disorders such as post traumatic stress disorder (PTSD) as well as
depression, headache, anxiety, cardiovascular disorders, and eating
disorders. Methods of treatment of such disorders and well as
packaged pharmaceutical compositions are also disclosed.
Inventors: |
Horvath, Raymond F;
(Guilford, CT) |
Correspondence
Address: |
Steven J. Sarussi
McDonnell Boehnen Hulbert & Berghoff
32nd Floor
300 S. Wacker Drive
Chicago
IL
60606
US
|
Assignee: |
Neurogen Corporation
|
Family ID: |
22450710 |
Appl. No.: |
09/930734 |
Filed: |
August 15, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09930734 |
Aug 15, 2001 |
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09561569 |
Apr 28, 2000 |
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6284766 |
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60131719 |
Apr 30, 1999 |
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Current U.S.
Class: |
514/267 ;
544/280 |
Current CPC
Class: |
A61P 25/04 20180101;
A61P 25/18 20180101; A61P 3/04 20180101; A61P 9/00 20180101; A61P
25/00 20180101; A61P 25/24 20180101; G01N 2333/5751 20130101; A61P
25/22 20180101; G01N 2333/726 20130101; C07D 487/04 20130101; A61P
25/30 20180101 |
Class at
Publication: |
514/261 ;
514/266; 544/277 |
International
Class: |
A61K 031/52; C07D
473/32 |
Claims
What is claimed is:
1. A compound of the formula: 25or pharmaceutically acceptable
salts thereof wherein Ar is phenyl, 1- or 2-naphthyl, 2-, 3-, or
4-pyridyl, 2-, 4- or 5-pyrimidinyl, each of which is mono-, di-, or
trisubstituted with halogen, trifluoromethyl, hydroxy, amino,
cyano, carboxamide, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.7 cycloalkyl, or amino(C.sub.1-C.sub.6)alkyl with the
proviso that at least one of the ortho or para positions of Ar is
substituted; R.sup.1 is hydrogen, halogen, trifluoromethyl,
carboxamide, carboxylate, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7
cycloalkyl, or (C.sub.1-C.sub.6 alkyl)--G.sup.1--R.sup.6 wherein
G.sup.1 is nitrogen, oxygen or sulfur and R.sup.6 is hydrogen,
C.sub.3-C.sub.7 cycloalkyl, or C.sub.1-C.sub.6 alkyl; R.sup.2 and
R.sup.3 together represent --(C.sub.0-C.sub.2)--G.sup.-
2-(C.sub.2-C.sub.4)-- wherein G.sup.2 is methylene, oxygen, sulfur
or NR.sup.7, wherein R.sup.7 is hydrogen, C.sub.3-C.sub.7
cycloalkyl, or C.sub.1-C.sub.6 alkyl; or R.sup.2 and R.sup.3 taken
together represent --CH.dbd.A--CH.dbd.CH-- wherein A is N or
CR.sup.8; R.sup.8 is --(C.sub.0-C.sub.6 alkyl)--Z; Z is
--CR.sup.9R.sup.9', NR.sup.9R.sup.9', OR.sup.9 or SR.sup.9; and
R.sup.9 and R.sup.9' independently represent hydrogen or
(C.sub.1-C.sub.6)alkyl; R.sup.4 and R.sup.5 are the same or
different and represent hydrogen, hydroxy, C.sub.1-C.sub.6
alkoxy(C.sub.1-C.sub.6)alkyl, hydroxy(C.sub.1-C.sub.6)alkyl,
C.sub.3-C.sub.7 cycloalkyl, or C.sub.1-C.sub.6 alkyl; or phenyl,
2-, 3-, or 4-pyridyl, 2- or 3-thienyl, or 2-, 4-, or 5-pyrimidinyl,
each of which is optionally mono- or disubstituted with halogen,
trifluoromethyl, hydroxy, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7
cycloalkyl, or C.sub.1-C.sub.6 alkoxy; or R.sup.4 and R.sup.5
together represent
--(C.sub.2-C.sub.3)--G.sup.3--(C.sub.1-C.sub.3)-- where G.sup.3 is
methylene, 1,2 phenylene, oxygen, sulfur or NR.sup.10; and R.sup.10
is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, phenyl, 2-,
3-, or 4-pyridyl, 2- or 3-thienyl, or 2-, 4- or 5-pyrimidyl.
2. A compound according to claim 1, wherein R.sub.2 and R.sub.3
together represent --CH.dbd.CR.sup.8--CH.dbd.CH--.
3. A compound according to claim 2, wherein R.sup.8 is hydrogen or
C.sub.1-C.sub.6 alkyl.
4. A compound according to claim 1, wherein Ar is phenyl mono-,
di-, or trisubstituted with halogen, trifluoromethyl, hydroxy,
amino, cyano, carboxamide, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkoxy, C.sub.3-C.sub.7 cycloalkyl, or amino(C.sub.1-C.sub.6)alkyl
with the proviso that at least one of the ortho positions of Ar is
substituted.
5. A compound according to claim 4, wherein Ar is phenyl
substituted in the 2 and 4 positions with methyl, fluoro or
chloro.
6. A compound according to claim 3, wherein Ar is
2,4,6-trimethylphenyl.
7. A compound according to claim 6, wherein R.sup.4 and R.sup.5
independently represent hydrogen, C.sub.1-C.sub.6
alkoxy(C.sub.1-C.sub.6) alkyl, C.sub.3-C.sub.7 cycloalkyl, or
C.sub.1-C.sub.6 alkyl.
8. A compound according to claim 2, wherein R.sup.8 is
--NR.sup.9R.sup.9'.
9. A compound according to claim 2, wherein R.sup.8 is
--(C.sub.1-C.sub.6 alkyl)--NR.sup.9R.sup.9'.
11. A compound according to claim 8, wherein Ar is
2,4,6-trimethylphenyl.
10. A compound according to claim 8, wherein R.sup.4 and R.sup.5
independently represent hydrogen, C.sub.1-C.sub.6
alkoxy(C.sub.1-C.sub.6) alkyl, C.sub.3-C.sub.7 cycloalkyl, or
C.sub.1-C.sub.6 alkyl.
12. A compound according to claim 1, wherein R.sup.2 and R.sup.3
together represent
--(C.sub.0-C.sub.2)CH.sub.2(C.sub.2-C.sub.4)--.
13. A compound according to claim 12, wherein Ar is
2,4,6-trimethylphenyl.
14. A compound according to claim 13, wherein R.sup.4 and R.sup.5
independently represent hydrogen, C.sub.1-C.sub.6
alkoxy(C.sub.1-C.sub.6) alkyl, C.sub.3-C.sub.7 cycloalkyl, or
C.sub.1-C.sub.6 alkyl.
15. A compound according to claim 1 which is
4-(N-Cyclopropylmethyl-N-prop-
yl)amino-2-methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidi-
no[4,5-b]indole.
16. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-2-methy-
l-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole.
17. A compound according to claim 1 which is
4-(N-Propyl-N-methyl)amino-2--
methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]in-
dole.
18. A compound according to claim 1 which is
4-(N-Cyclopropylmethyl-N-prop-
yl)amino-2-methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole.
19. A compound according to claim 1 which
4-(N,N-Dipropyl)amino-2-methyl-9-
-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole.
20. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-6-ethyl-
amino-2-methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole.
21. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-6-dieth-
ylamino-2-methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole.
22. A compound according to claim 1 which is
4-Bis(2-methoxyethyl)amino-2--
methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]in-
dole.
23. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-2-methy-
l-9-(4-bromo-2,6-dimethylphenyl)-9H-pyrimidino[4,5-b]indole.
24. A compound according to claim 1 which is
4-Bis(2-methoxyethyl)amino-2--
methyl-9-(4-bromo-2,6-dimethylphenyl)-9H-pyrimidino[4,5-b]indole.
25. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-2-methy-
l-9-(4-chloro-2-methylphenyl)-9H-pyrimidino [4,5-b]l indole.
26. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-2-methy-
l-9-(4-cyano-2,6-dimethylphenyl)-9H-pyrimidino [4,5-b]indole.
27. A compound according to claim 1 which is
4-Bis(2-methoxyethyl)amino-2--
methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole.
28. A compound according to claim 1 which is
4-(N-Cyclopropylmethyl-N-prop-
yl)amino-6-ethylamino-2-methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-
-b]indole.
29. A compound according to claim 1 which is
4-(N-(2-Hydroxyethyl)-N-Cyclo-
propylmethyl)amino-2-methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]-
indole.
30. A compound according to claim 1 which is
4-(N-(2-Hydroxyethyl)-N-Cyclo-
propyl)amino-2-methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole-
.
31. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-2-methy-
l-(2,4,6-trimethylphenyl)-5,6,7-trihydrocyclopenta[2,1-d]-8H-pyrimidino[4,-
5-b]indole
32. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-2-methy-
l-9-(2,6-dimethylphenyl)-9H-pyrimidino[4,5-b]indole.
33. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-2-methy-
l-9-(4-aminomethyl-2,6-dimethylphenyl)-9H-pyrimidino[4,5-b]indole.
34. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-2-methy-
l-9-(2,6-dimethyl-4-(methylaminomethyl)phenyl)-9H-pyrimidino
[4,5-b]indole.
35. A compound according to claim 1 which is
4-(N-Cyclopropylmethyl-N-prop-
yl)amino-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]-
indole.
36. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-9-(2,4,-
6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole.
37. A compound according to claim 1 which is
4-(N-Butyl-N-methyl)amino-9-(-
2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole.
38. A compound according to claim 1 which is
4-(N-Propyl)amino-8-(2,4,6-tr-
imethylphenyl)-5,6,7-trihydrocyclopenta[2,1-d]-8H-pyrimidino[4,5-b]indole.
39. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-8-(2,4,-
6-trimethylphenyl)-5,6,7-trihydrocyclopenta[2,1-d]-8H-pyrimidino[4,5-b]ind-
ole.
40. A compound according to claim 1 which is Ethyl
4-(dipropylamino)-9-(2,-
4,6-trimethylphenyl)-5,6,7,8-tetrahydropyrimidino[4,5-b]indole-2-carboxyla-
te.
41. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-2-carbo-
xyamido-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]i-
ndole.
42. A compound according to claim 1 which is
4-(N,N-Dipropyl)amino-2-amino-
methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]in-
dole.
43. A compound according to claim 1 which is
4-(N-Cyclopropylmethylamino)--
2-methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole.
44.
4-(N,N-Dibutylamino)-2-methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrah-
ydro-9H-pyrimidino[4,5-b]indole.
45.
4-(N,N-Dibutylamino)-2-methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrah-
ydro-9H-pyrimidino[4,5-b]indole.
46. A compound according to claim 1 which is
4-(N-(2-(4-Chlorophenyl)ethyl-
)amino)-2-methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidin-
o[4,5-b]indole.
47. A compound according to claim 1 which is
4-(N-(2-(4-Chlorophenyl)ethyl-
)amino)-2-methyl-9-(2,4-dimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4-
,5-b]indole.
48. A compound according to claim 1 which is
4-(N-(4-Chlorophenyl)methylam-
ino)-2-methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4-
,5-b]indole.
49. A compound according to claim 1 which is
4-(N-(2-Methylphenyl)methylam-
ino)-2-methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4-
,5-b]indole.
50. A compound according to claim 1 which is
4-(N-(3-Trifluromethylphenyl)-
methylamino)-2-methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyri-
midino[4,5-b]indole.
51. A compound according to claim 1 which is
(N,N-Dipropylamino)-2-methyl--
6-dimethylamino-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole.
52. A compound according to claim 1, which is
4-(N-Butyl-N-methyl)amino-9--
(2,4,6-trimethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole.
53. A pharmaceutical composition comprising a compound according to
claim 1 and at least one pharmaceutically acceptable carrier or
excipient.
54. A process for preparing a compound according to claim 1.
55. A packaged pharmaceutical composition comprising the
pharmaceutical composition of claim 53 in a container and
instructions for using the composition to treat a patient in need
thereof.
56. The packaged pharmaceutical composition of claim 55, wherein
said patient is suffering from stress, posttraumatic stress
disorder, anxiety, depression, cardiovascular disease, headache,
obesity or an eating disorder.
57. A method for localizing CRF receptors in tissue section samples
comprising: contacting with a sample of tissue a
detectably-labelled compound of claim 1 under conditions that
permit binding of the compound to the sample of tissue; washing the
tissue sample to remove unbound compound; and detecting the bound
compound.
58. A method for the treatment or prevention of physiological
disorders associated with excess of or insufficient amount of CRF,
which method comprises administration to a patient in need thereof
an effect amount of a compound according to claim 1.
59. A method of inhibiting the binding of CRF to the CRF1 receptor,
which method comprises contacting, in the presence of CRF, a
solution comprising a compound of claim 1, with cells expressing
the CRF1 receptor, wherein the compound is present in the solution
at a concentration sufficient to reduce levels of CRF binding to
IMR32 cells in vitro.
60. A method for altering the signal-transducing activity of a cell
surface CRF1 receptor, said method comprising contacting cells
expressing such a receptor with a solution comprising a compound
according to claim 1, wherein the compound is present in the
solution at a concentration sufficient to reduce levels of CRF
binding to IMR32 cells in vitro.
61. A compound according to claim 1 wherein in a standard assay of
CRF binding the compound exhibits an IC.sub.50 of 1 micromolar or
less.
62. A compound according to claim 1 wherein in a standard assay of
CRF binding the compound exhibits an IC.sub.50 of 100 nanomolar or
less.
63. A compound according to claim 1 wherein in a standard assay of
CRF binding the compound exhibits an IC.sub.50 of 10 nanomolar or
less.
64. A method for treating stress, posttraumatic stress disorder,
anxiety or depression which comprises administering an effective
amount of a compound according to claim 1 to a patient in need
thereof.
65. A method for treating obesity or eating disorders which
comprises administering an effective amount of a compound according
to claim 1 to a patient in need thereof.
66. A method for treating cardiovascular disorders which comprises
administering an effective amount of a compound according to claim
1 to a patient in need thereof.
67. A method for treating headache which comprises administering an
effective amount of a compound according to claim 1 to a patient in
need thereof.
Description
[0001] This application claims priority from provisional
application no. 60/131,719, filed Apr. 30, 1999, which is
incorporated herein by reference in its entirety.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] This invention relates to 9H-pyrimido[4,5-b]indole
derivatives that modulate cell surface receptors, especially
corticotropin-releasing factor 1 (CRF1) receptors. The invention
also relates to 9H-pyrimido[4,5-b]indole derivatives that bind
selectively to CRF1 receptors. In a further aspect the invention
includes pharmaceutical compositions comprising such compounds and
the use of such compounds in treatment of psychiatric disorders and
neurological diseases, including major depression, anxiety-related
disorders, post-traumatic stress disorder (PTSD), supranuclear
palsy and feeding disorders, as well as treatment of immunological,
cardiovascular or heart-related diseases and colonic
hypersensitivity associated with psychopathological disturbance and
stress. Additionally this invention relates to the use such
compounds as probes for the localization of CRF1 receptors in cells
and tissues.
[0004] 2. Description of the Related Art
[0005] Corticotropin releasing factor (CRF), a 41 amino acid
peptide, is the primary physiological regulator of
proopiomelanocortin (POMC) derived peptide secretion from the
anterior pituitary gland. In addition to its endocrine role at the
pituitary gland, immunohistochemical localization of CRF has
demonstrated that the hormone has a broad extrahypothalamic
distribution in the central nervous system and produces a wide
spectrum of autonomic, electrophysiological and behavioral effects
consistent with a neurotransmitter or neuromodulator role in brain.
There is also evidence that CRF plays a significant role in
integrating the response of the immune system to physiological,
psychological, and immunological stressors.
[0006] Clinical data provide evidence that CRF has a role in
psychiatric disorders and neurological diseases including
depression, anxiety-related disorders and feeding disorders. A role
for CRF has also been postulated in the etiology and
pathophysiology of Alzheimer's disease, Parkinson's disease,
Huntington's disease, progressive supranuclear palsy and
amyotrophic lateral sclerosis as they relate to the dysfunction of
CRF neurons in the central nervous system.
[0007] In affective disorder, or major depression, the
concentration of CRF is significantly increased in the cerebral
spinal fluid (CSF) of drug-free individuals. Furthermore, the
density of CRF receptors is significantly decreased in the frontal
cortex of suicide victims, consistent with a hypersecretion of CRF.
In addition, there is a blunted adrenocorticotropin (ACTH) response
to CRF (i.v. administered) observed in depressed patients.
Preclinical studies in rats and non-human primates provide
additional support for the hypothesis that hypersecretion of CRF
may be involved in the symptoms seen in human depression. There is
also preliminary evidence that tricyclic antidepressants can alter
CRF levels and thus modulate the numbers of CRF receptors in
brain.
[0008] CRF has also been implicated in the etiology of
anxiety-related disorders. CRF produces anxiogenic effects in
animals and interactions between benzodiazepine/non-benzodiazepine
anxiolytics and CRF have been demonstrated in a variety of
behavioral anxiety models. Preliminary studies using the putative
CRF receptor antagonist -helical ovine CRF (9-41) in a variety of
behavioral paradigms demonstrate that the antagonist produces
"anxiolytic-like" effects that are qualitatively similar to the
benzodiazepines. Neurochemical, endocrine and receptor binding
studies have all demonstrated interactions between CRF and
benzodiazepine anxiolytics providing further evidence for the
involvement of CRF in these disorders. Chlordiazepoxide attenuates
the "anxiogenic" effects of CRF in both the conflict test and in
the acoustic startle test in rats. The benzodiazepine receptor
antagonist Ro 15-1788, which was without behavioral activity alone
in the operant conflict test, reversed the effects of CRF in a
dose-dependent manner, while the benzodiazepine inverse agonist FG
7142 enhanced the actions of CRF.
[0009] CRF has also been implicated in the pathogeneisis of certain
immunological, cardiovascular or heart-related diseases such as
hypertension, tachycardia and congestive heart failure, stroke and
osteoporosis, as well as in premature birth, psychosocial dwarfism,
stress-induced fever, ulcer, diarrhea, post-operative ileus and
colonic hypersensitivity associated with psychopathological
disturbance and stress.
[0010] The mechanisms and sites of action through which
conventional anxiolytics and antidepressants produce their
therapeutic effects remain to be fully elucidated. It has been
hypothesized however, that they are involved in the suppression of
CRF hypersecretion that is observed in these disorders. Of
particular interest are that preliminary studies examining the
effects of a CRF receptor antagonist peptide ( -helical
CRF.sub.9-41) in a variety of behavioral paradigms have
demonstrated that the CRF antagonist produces "anxiolytic-like"
effects qualitatively similar to the benzodiazepines.
[0011] Certain pyrido[2,3-b]indoles and pyrimido[4,5-b]indoles have
been described. See, for example, Arzneim.-Forsch. 1978, 28,
1056-65) and Khim. Geterotsikl. Soedin. 1991, 6, 852).
SUMMARY OF THE INVENTION
[0012] The invention provides compounds of Formula I (shown below),
and pharmaceutical compositions comprising compounds of Formula I.
Such compounds bind to cell surface receptors, preferably G-coupled
protein receptors, especially CRF receptors and most preferably
CRF1 receptors. Preferred compounds of the invention exhibit high
affinity for CRF1 receptors. Additionally, preferred compounds of
the invention also exhibit high specificity for CRF1 receptors.
[0013] Preferred compounds of the present invention exhibit
activity as corticotropin releasing factor receptor antagonists and
appear to suppress the anxiogenic effects of CRF hypersecretion.
The invention also provides methods of using compounds of Formula I
for the suppression of CRF hypersecretion and for the treatment of
anxiogenic and stress-related disorders.
[0014] The invention further comprises methods of treating patients
suffering from certain disorders that are responsive to modulation
of CRF1 receptors with an effective amount of a compound of the
invention. These disorders include CNS disorders, particularly
affective disorders, anxiety disorders, stress-related disorders,
including post traumatic stress disorder (PTSD) as well as
depression, headache, eating disorders and substance abuse. In
another aspect, the invention provides methods for preventing such
disorders, which methods comprise administration to a patient an
effective amount of a compound of Formula I.
[0015] Treatment of human patients suffering from such disorders as
well as other animals domesticated companion animals (pets) or
livestock animals in encompassed by the invention.
[0016] In another aspect, the invention provides pharmaceutical
compositions comprising compounds of Formula I or the
pharmaceutically acceptable salts or solvates thereof. The
invention also includes packaged pharmaceutical compositions of
compounds, together with instructions for use.
[0017] Additionally this invention relates to the use of the
compounds of the invention (particularly labeled compounds of this
invention) as probes for the localization of receptors in cells and
tissues and as standards and reagents for use in determining the
receptor-binding characteristics of test compounds. Labeled
compounds of the invention may be used for in vitro studies such as
autoradiography of tissue sections or for in vivo methods, e.g. PET
or SPECT scanning. Particularly, preferred compounds of the
invention are useful as standards and reagents in determining the
ability of a potential pharmaceutical to bind to the CRF1
receptor.
[0018] The invention also provides methods of inhibiting the
binding of CRF to CRF receptors and to methods of altering the
signal-transducing activity of CRF receptors.
[0019] In yet another aspect, the invention provides intermediates
useful in the preparation of the compounds of Formula I.
[0020] Accordingly, a broad embodiment of the invention is directed
to compounds of Formula I: 2
[0021] or pharmaceutically acceptable salts thereof, wherein: Ar,
R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are defined
below.
DETAILED DESCRIPTION OF THE INVENTION
[0022] The invention provides compounds of Formula I: 3
[0023] or pharmaceutically acceptable salts thereof, wherein:
[0024] Ar is phenyl, 1- or 2-naphthyl, 2-, 3-, or 4-pyridyl, 2-, 4-
or 5-pyrimidinyl, each of which is mono-, di-, or trisubstituted
with halogen, trifluoromethyl, hydroxy, amino, cyano, carboxamide,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, C.sub.3-C.sub.7
cycloalkyl, or amino (C.sub.1-C.sub.6)alkyl with the proviso that
at least one of the ortho or para positions of Ar is
substituted;
[0025] R.sup.1 is hydrogen, halogen, trifluoromethyl, carboxamide,
carboxylate, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7 cycloalkyl, or
(C.sub.1-C.sub.6 alkyl)-G.sup.1-R.sup.6 wherein G.sup.1 is
nitrogen, oxygen or sulfur and R.sup.6 is hydrogen, C.sub.3-C.sub.7
cycloalkyl, or C.sub.1-C.sub.6 alkyl;
[0026] R.sup.2 and R.sup.3 together represent
--(C.sub.0-C.sub.2)--G.sup.2- --(C.sub.2-C.sub.4)- wherein G.sup.2
is methylene, oxygen, sulfur or NR.sup.7, wherein R.sup.7 is
hydrogen, C.sub.3-C.sub.7 cycloalkyl, or C.sub.1-C.sub.6 alkyl;
or
[0027] R.sup.2 and R.sup.3 taken together represent
--CH.dbd.A--CH.dbd.CH-- wherein
[0028] A is N or CR.sup.8;
[0029] R.sup.8 is --(C.sub.0-C.sub.6 alkyl)--Z;
[0030] where
[0031] Z is --CR.sup.9R.sup.9', NR.sup.9R.sup.9', OR.sup.9 or
SR.sup.9; and
[0032] R.sup.9 and R.sup.9' independently represent hydrogen or
(C.sub.1-C.sub.6)alkyl;
[0033] R.sup.4 and R.sup.5 are the same or different and represent
hydrogen, hydroxy, C.sub.1-C.sub.6 alkoxy(C.sub.1-C.sub.6)alkyl,
hydroxy(C.sub.1-C.sub.6)alkyl, C.sub.3-C.sub.7 cycloalkyl, or
C.sub.1-C.sub.6 alkyl; or
[0034] phenyl, 2-, 3-, or 4-pyridyl, 2- or 3-thienyl, or 2-, 4-, or
5-pyrimidinyl, each of which is optionally mono- or disubstituted
with halogen, trifluoromethyl, hydroxy, C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.7 cycloalkyl, or C.sub.1-C.sub.6 alkoxy; or
[0035] R.sup.4 and R.sup.5 together represent
--(C.sub.2-C.sub.3)--G.sup.3- --(C.sub.1-C.sub.3)-- where
[0036] G.sup.3 is methylene, 1,2-phenylene, oxygen, sulfur or
NR.sup.10; and
[0037] R.sup.10 is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.7
cycloalkyl, phenyl, 2-, 3-, or 4-pyridyl, 2- or 3-thienyl, or 2-,
4- or 5-pyrimidyl.
[0038] Preferred compounds of Formula I include those where Ar is
phenyl mono-, di-, or trisubstituted with halogen, trifluoromethyl,
hydroxy, amino, cyano, carboxamide, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxy, C.sub.3-C.sub.7 cycloalkyl, or
amino(C.sub.1-C.sub.6)alkyl with the proviso that at least one of
the ortho positions of Ar is substituted. Other preferred compounds
of Formula I are those where R.sub.1 is lower alkyl, more
preferably C.sub.1-C.sub.3 alkyl, and most preferably methyl. More
preferred compounds of Formula I include those where Ar is phenyl
substituted in the 2 and 4 positions with methyl, fluoro or chloro,
or phenyl trisubstituted in the 2, 4, and 6-positions with
C.sub.1-C.sub.2 alkyl, more preferably methyl. Other preferred
compounds of Formula I are those where R.sup.4 and R.sup.5
independently represent hydrogen, C.sub.1-C.sub.6
alkoxy(C.sub.1-C.sub.6) alkyl, C.sub.3-C.sub.7 cycloalkyl, or
C.sub.1-C.sub.6 alkyl. Still other preferred compounds of the
invention are those where R.sub.2 is C.sub.1-C.sub.2 alkyl,
C.sub.1-C.sub.2 alkyl carboxylate, or amino (C.sub.1-C.sub.2)
alkyl.
[0039] A preferred group of compounds of Formula I are those
wherein R.sub.2 and R.sub.3 together represent
--CH.dbd.CR.sup.8--CH.dbd.CH--. Such compounds are represented by
Formula II: 4
[0040] where the substituents are defined as for Formula I.
[0041] Preferred compounds of Formula II are those where wherein
R.sup.8 is hydrogen or C.sub.1-C.sub.6 alkyl. Such compounds are
hereinafter referred to as compounds of Formula II-A. Preferred
compounds of Formula II-A include those where Ar is phenyl
substituted in the 2 and 4 positions with methyl, fluoro or chloro
or Ar is 2,4,6-trimethylphenyl. Other preferred compounds of
Formula II-A are those where R.sub.1 is lower alkyl, more
preferably C.sub.1-C.sub.3 alkyl, and most preferably methyl. More
preferably, compounds of Formula II-A are those where Ar is
2,4,6-trimethylphenyl and R.sub.1 is lower alkyl, more preferably
C.sub.1-C.sub.3 alkyl, and most preferably methyl.
[0042] Particularly preferred compounds of Formula II-A include
those where R.sup.8 is hydrogen or C.sub.1-C.sub.6 alkyl, Ar is
2,4,6-trimethylphenyl; and R.sup.4 and R.sup.5 independently
represent hydrogen, C.sub.1-C.sub.6 alkoxy(C.sub.1-C.sub.6) alkyl,
C.sub.3-C.sub.7 cycloalkyl, or C.sub.1-C.sub.6 alkyl.
[0043] Other preferred compounds of Formula II are those where
R.sup.8 is --(C.sub.0-C.sub.6 alkyl)--OR.sup.9 or
--(C.sub.0-C.sub.6 alkyl)--NR.sup.9R.sup.9'; and R.sup.9 and
R.sup.9' are independently hydrogen or (C.sub.1-C.sub.6)alkyl. Such
compounds are designated as compounds of Formula II-B herein. More
preferred are compounds of II-B are where R.sup.8 is
--NR.sup.9R.sup.9'. Particularly preferred compounds of II-B are
those where at least one of R.sup.9 and R.sup.9' is C.sub.1-C.sub.6
alkyl, preferably methyl, ethyl, or propyl. The most preferred
compounds of II-B are those where Ar is 2,4,6-trimethylphenyl.
Other most preferred compounds of Formula II-B are those where
R.sup.4 and R.sup.5 independently represent hydrogen,
C.sub.1-C.sub.6 alkoxy(C.sub.1-C.sub.6) alkyl, C.sub.3-C.sub.7
cycloalkyl, or C.sub.1-C.sub.6 alkyl.
[0044] Other preferred compounds of Formula II-B are those where
R.sub.1 is lower alkyl, more preferably C.sub.1-C.sub.3 alkyl, and
most preferably methyl. More preferably, compounds of Formula II-B
are those where Ar is 2,4,6-trimethylphenyl and R.sub.1 is lower
alkyl, more preferably C.sub.1-C.sub.3 alkyl, and most preferably
methyl.
[0045] Other preferred compounds of Formula I are those where
R.sup.2 and R.sup.3 together represent
--(C.sub.0-C.sub.2)CH.sub.2(C.sub.2-C.sub.4)--- . Such compounds
are represented by Formula III. 5
[0046] Preferred compounds of Formula III are those where
--(C.sub.0-C.sub.2)CH.sub.2(C.sub.2-C.sub.4)-- and the carbon atoms
at the 5 and 6 positions of the pyrrolo[2,3-d]pyrimidine form a
five- or six-membered ring. Examples of such compounds have the
following general formulas: 6
[0047] Since C.sub.2, C.sub.3, and C.sub.4 include branched
alkylene groups, the compounds of these formulas may be substituted
on the unsaturated ring (i.e., the non-nitrogen-containing ring)
with C.sub.1-C.sub.6 alkyl within the definition of
--(C.sub.0-C.sub.2)CH.sub.- 2(C.sub.2-C.sub.4)--. Preferred
compounds of Formula III include those wherein Ar is
2,4,6-trimethylphenyl. Particularly preferred compounds of Formula
III are those where R.sup.4 and R.sup.5 independently represent
hydrogen, C.sub.1-C.sub.6 alkoxy(C.sub.1-C.sub.6) alkyl,
C.sub.3-C.sub.7 cycloalkyl, or C.sub.1-C.sub.6 alkyl.
[0048] Other preferred compounds of Formula III are those where
R.sub.1 is lower alkyl, more preferably C.sub.1-C.sub.3 alkyl, and
most preferably methyl. More preferably, compounds of Formula III
are those where Ar is 2,4,6-trimethylphenyl and R.sub.1 is lower
alkyl, more preferably C.sub.1-C.sub.3 alkyl, and most preferably
methyl. Particularly preferred compounds of Formula III are those
where Ar is 2,4,6-trimethylphenyl, R.sub.1 is lower alkyl,
preferably methyl, and R.sup.4 and R.sup.5 independently represent
hydrogen, C.sub.1-C.sub.6 alkoxy(C.sub.1-C.sub.6) alkyl,
C.sub.3-C.sub.7 cycloalkyl, or C.sub.1-C.sub.6 alkyl.
[0049] In certain situations, the compounds of Formula I may
contain one or more asymmetric carbon atoms, so that the compounds
can exist in different stereoisomeric forms. These compounds can
be, for example, racemates or optically active forms. In these
situations, the single enantiomers, i.e., optically active forms,
can be obtained by asymmetric synthesis or by resolution of the
racemates. Resolution of the racemates can be accomplished, for
example, by conventional methods such as crystallization in the
presence of a resolving agent, or chromatography, using, for
example a chiral HPLC column.
[0050] Representative compounds of the present invention, which are
encompassed by Formula I, include, but are not limited to the
compounds in Table 1 and their pharmaceutically acceptable acid
addition salts. In addition, if the compound of the invention is
obtained as an acid addition salt, the free base can be obtained by
basifying a solution of the acid salt. Conversely, if the product
is a free base, an addition salt, particularly a pharmaceutically
acceptable addition salt, may be produced by dissolving the free
base in a suitable organic solvent and treating the solution with
an acid, in accordance with conventional procedures for preparing
acid addition salts from base compounds.
[0051] Non-toxic pharmaceutical salts include salts of acids such
as hydrochloric, phosphoric, hydrobromic, sulfuric, sulfinic,
formic, toluenesulfonic, methanesulfonic, nitric, benzoic, citric,
tartaric, maleic, hydroiodic, alkanoic such as acetic,
HOOC--(CH.sub.2).sub.n--COOH where n is 0-4, and the like. Those
skilled in the art will recognize a wide variety of non-toxic
pharmaceutically acceptable addition salts.
[0052] The present invention also encompasses the acylated prodrugs
of the compounds of Formula I. Those skilled in the art will
recognize various synthetic methodologies which may be employed to
prepare non-toxic pharmaceutically acceptable addition salts and
acylated prodrugs of the compounds encompassed by Formula I.
[0053] Where a compound exists in various tautomeric forms, the
invention is not limited to any one of the specific tautomers. The
invention includes all tautomeric forms of a compound.
[0054] By "C.sub.1-C.sub.6 alkyl" or "lower alkyl" in the present
invention is meant straight or branched chain alkyl groups having
1-6 carbon atoms, such as, for example, methyl, ethyl, propyl,
isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, 2-pentyl,
isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl.
Preferred C.sub.1-C.sub.6 alkyl groups are methyl, ethyl, propyl,
and butyl
[0055] Preferred C.sub.3-C.sub.7 cycloalkyl groups herein are
cyclopropyl and cyclopropylmethyl.
[0056] By "C.sub.1-C.sub.6 alkoxy" or "lower alkoxy" in the present
invention is meant straight or branched chain alkoxy groups having
1-6 carbon atoms, such as, for example, methoxy, ethoxy, propoxy,
isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentoxy, 2-pentyl,
isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, and
3-methylpentoxy.
[0057] By the term "halogen" in the present invention is meant
fluorine, bromine, chlorine, and iodine.
[0058] As used herein, C.sub.0 represents a bond.
[0059] When the terms C.sub.1, C.sub.2, C.sub.3, and C.sub.4 are
used the context of a "C.sub.0-C.sub.2", "C.sub.1-C.sub.3",
"C.sub.1-C.sub.3", or "C.sub.2-C.sub.4" group, the terms represent
alkylene groups having 1, 2, 3, or 4 carbon atoms respectively. The
alkylene groups may be straight or branched chain groups. For
example, C.sub.3 includes groups of the formulas: 7
[0060] Examples of C.sub.4 groups are: 8
[0061] Examples of compounds of the invention where R.sup.2 and
R.sup.3 together represent groups of the formula
--(C.sub.0-C.sub.2)--G.sup.2--(C- .sub.2-C.sub.4)-- include the
following: 9
[0062] The alkylene groups in the
--(C.sub.0-C.sub.2)--G.sup.2-(C.sub.2-C.- sub.4)-- group, i.e.,
"C.sub.1", "C.sub.2", "C.sub.3", and "C.sub.4" groups, may,
provided there is a sufficient number of carbon atoms, be straight
or branched chain alkyl groups. Thus, the invention encompasses
compounds having the following general structures: 10
[0063] Representative compounds of the invention are shown in Table
1.
1TABLE 1 1 11 2 12 3 13 4 14 5 15 6 16
[0064] The present invention also encompasses methods for
inhibiting the binding of CRF to the CRF1 receptor which methods
involve contacting a compound of the invention with cells
expressing CRF1 receptors, wherein the compound is present at a
concentration sufficient to inhibit CRF binding to cells expressing
CRF1 receptors in vitro. These methods include inhibiting the
binding of CRF to CRF1 receptors in vivo, e.g., in a patient using
an amount of a compound of Formula I that would be sufficient to
inhibit the binding of CRF to CRF1 receptors in vitro. The cells
expressing CRF1 receptors in vitro may either naturally express
CRF1 receptors, e.g. IMR32 cells, or may recombinantly express
cloned CRF1 receptors. The amount of a compound that would be
sufficient to inhibit the binding of CRF to the CRF1 receptor may
be readily determined via CRF1 receptor binding assays, such as the
assay described in Example 6.
[0065] The present invention also pertains to methods for altering
the signal-transducing activity of CRF1 receptors, said method
comprising exposing cells expressing such receptors to an effective
amount of a compound of the invention. These methods include
altering the signal-transducing activity of CRF1 receptors in vivo,
e.g., in a patient using an amount of a compound of Formula I that
would be sufficient to alter the signal-transducing activity of
CRF1 receptors in vitro. The amount of a compound that would be
sufficient to alter the signal-transducing activity of CRF1
receptors may be determined via an CRF1 receptor signal
transduction assay, such as the assay described in Example 7.
[0066] The compounds of the invention and labeled derivatives
thereof are also useful as standards and reagents in determining
the ability of a potential pharmaceutical to bind to the CRF1
receptor.
[0067] Labeled derivatives of the compounds provided by this
invention are also useful as radiotracers for positron emission
tomography (PET) imaging or for single photon emission computerized
tomography (SPECT).
[0068] The invention also pertains to the use of compounds of
general Formula I in the treatment of various stress-related
disorders. The interaction of compounds of the invention with CRF
receptors is shown in the examples.
[0069] The compounds of general Formula I may be administered
orally, topically, parenterally, by inhalation or spray or rectally
in dosage unit formulations containing conventional non-toxic
pharmaceutically acceptable carriers, adjuvants and vehicles. Oral
administration in the form of a pill, capsule, elixir, syrup,
lozenge, troche, or the like is particularly preferred. The term
parenteral as used herein includes subcutaneous injections,
intradermal, intravascular (e.g., intravenous), intramuscular,
spinal, intrathecal injection or like injection or infusion
techniques. In addition, there is provided a pharmaceutical
formulation comprising a compound of general Formula I and a
pharmaceutically acceptable carrier. One or more compounds of
general Formula I may be present in association with one or more
non-toxic pharmaceutically acceptable carriers and/or diluents
and/or adjuvants and if desired other active ingredients. The
pharmaceutical compositions containing compounds of general Formula
I may be in a form suitable for oral use, for example, as tablets,
troches, lozenges, aqueous or oily suspensions, dispersible powders
or granules, emulsion, hard or soft capsules, or syrups or
elixirs.
[0070] Compositions intended for oral use may be prepared according
to any method known to the art for the manufacture of
pharmaceutical compositions and such compositions may contain one
or more agents selected from the group consisting of sweetening
agents, flavoring agents, coloring agents and preserving agents in
order to provide pharmaceutically elegant and palatable
preparations. Tablets contain the active ingredient in admixture
with non-toxic pharmaceutically acceptable excipients that are
suitable for the manufacture of tablets. These excipients may be
for example, inert diluents, such as calcium carbonate, sodium
carbonate, lactose, calcium phosphate or sodium phosphate;
granulating and disintegrating agents, for example, corn starch, or
alginic acid; binding agents, for example starch, gelatin or
acacia, and lubricating agents, for example magnesium stearate,
stearic acid or talc. The tablets may be uncoated or they may be
coated by known techniques to delay disintegration and absorption
in the gastrointestinal tract and thereby provide a sustained
action over a longer period. For example, a time delay material
such as glyceryl monostearate or glyceryl distearate may be
employed.
[0071] Formulations for oral use may also be presented as hard
gelatin capsules wherein the active ingredient is mixed with an
inert solid diluent, for example, calcium carbonate, calcium
phosphate or kaolin, or as soft gelatin capsules wherein the active
ingredient is mixed with water or an oil medium, for example peanut
oil, liquid paraffin or olive oil.
[0072] Aqueous suspensions contain the active materials in
admixture with excipients suitable for the manufacture of aqueous
suspensions. Such excipients are suspending agents, for example
sodium carboxymethylcellulose, methylcellulose,
hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone,
gum tragacanth and gum acacia; dispersing or wetting agents may be
a naturally-occurring phosphatide, for example, lecithin, or
condensation products of an alkylene oxide with fatty acids, for
example polyoxyethylene stearate, or condensation products of
ethylene oxide with long chain aliphatic alcohols, for example
heptadecaethyleneoxycetanol, or condensation products of ethylene
oxide with partial esters derived from fatty acids and a hexitol
such as polyoxyethylene sorbitol monooleate, or condensation
products of ethylene oxide with partial esters derived from fatty
acids and hexitol anhydrides, for example polyethylene sorbitan
monooleate. The aqueous suspensions may also contain one or more
preservatives, for example ethyl, or n-propyl p-hydroxybenzoate,
one or more coloring agents, one or more flavoring agents, and one
or more sweetening agents, such as sucrose or saccharin.
[0073] Oily suspensions may be formulated by suspending the active
ingredients in a vegetable oil, for example arachis oil, olive oil,
sesame oil or coconut oil, or in a mineral oil such as liquid
paraffin. The oily suspensions may contain a thickening agent, for
example beeswax, hard paraffin or cetyl alcohol. Sweetening agents
such as those set forth above, and flavoring agents may be added to
provide palatable oral preparations. These compositions may be
preserved by the addition of an anti-oxidant such as ascorbic
acid.
[0074] Dispersible powders and granules suitable for preparation of
an aqueous suspension by the addition of water provide the active
ingredient in admixture with a dispersing or wetting agent,
suspending agent and one or more preservatives. Suitable dispersing
or wetting agents and suspending agents are exemplified by those
already mentioned above. Additional excipients, for example
sweetening, flavoring and coloring agents, may also be present.
[0075] Pharmaceutical compositions of the invention may also be in
the form of oil-in-water emulsions. The oily phase may be a
vegetable oil, for example olive oil or arachis oil, or a mineral
oil, for example liquid paraffin or mixtures of these. Suitable
emulsifying agents may be naturally-occurring gums, for example gum
acacia or gum tragacanth, naturally-occurring phosphatides, for
example soy bean, lecithin, and esters or partial esters derived
from fatty acids and hexitol, anhydrides, for example sorbitan
monooleate, and condensation products of the said partial esters
with ethylene oxide, for example polyoxyethylene sorbitan
monooleate. The emulsions may also contain sweetening and flavoring
agents.
[0076] Syrups and elixirs may be formulated with sweetening agents,
for example glycerol, propylene glycol, sorbitol or sucrose. Such
formulations may also contain a demulcent, a preservative and
flavoring and coloring agents. The pharmaceutical compositions may
be in the form of a sterile injectable aqueous or oleaginous
suspension. This suspension may be formulated according to the
known art using those suitable dispersing or wetting agents and
suspending agents which have been mentioned above. The sterile
injectable preparation may also be sterile injectable solution or
suspension in a non-toxic parentally 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 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 may be
employed including synthetic mono- or diglycerides. In addition,
fatty acids such as oleic acid find use in the preparation of
injectables.
[0077] The compounds of general Formula I may also be administered
in the form of suppositories, e.g., for rectal administration of
the drug. These compositions can be prepared by mixing the drug
with a suitable non-irritating excipient that is solid at ordinary
temperatures but liquid at the rectal temperature and will
therefore melt in the rectum to release the drug. Such materials
are cocoa butter and polyethylene glycols.
[0078] Compounds of general Formula I may be administered
parenterally in a sterile medium. The drug, depending on the
vehicle and concentration used, can either be suspended or
dissolved in the vehicle. Advantageously, adjuvants such as local
anesthetics, preservatives and buffering agents can be dissolved in
the vehicle.
[0079] Dosage levels of the order of from about 0.1 mg to about 140
mg per kilogram of body weight per day are useful in the treatment
of the above-indicated conditions (about 0.5 mg to about 7 g per
patient per day). The amount of active ingredient that may be
combined with the carrier materials to produce a single dosage form
will vary depending upon the host treated and the particular mode
of administration. Dosage unit forms will generally contain between
from about 1 mg to about 500 mg of an active ingredient.
[0080] Frequency of dosage may also vary depending on the compound
used and the particular disease treated. However, for treatment of
most disorders, a dosage regimen of 4 times daily or less is
preferred. For the treatment of anxiety, depression, or stress
disorders a dosage regimen of 1 or 2 times daily is particularly
preferred.
[0081] It will be understood, however, that the specific dose level
for any particular patient will depend upon a variety of factors
including the activity of the specific compound employed, the age,
body weight, general health, sex, diet, time of administration,
route of administration, and rate of excretion, drug combination
and the severity of the particular disease undergoing therapy.
[0082] Preferred compounds of the invention will have certain
pharmacological properties. Such properties include, but are not
limited to oral bioavailability, low toxicity, low serum protein
binding and desirable in vitro and in vivo half-lifes. Penetration
of the blood brain barrier for compounds used to treat CNS
disorders is necessary, while low brain levels of compounds used to
treat peripheral disorders are often preferred.
[0083] Assays may be used to predict these desirable
pharmacological properties. Assays used to predict bioavailability
include transport across human intestinal cell monolayers,
including Caco-2 cell monolayers. Toxicity to cultured hepatocyctes
may be used to predict compound toxicity. Penetration of the blood
brain barrier of a compound in humans may be predicted from the
brain levels of the compound in laboratory animals given the
compound intravenously.
[0084] Serum protein binding may be predicted from albumin binding
assays. Such assays are described in a review by Oravcov, et al.
(Journal of Chromatography B (1996) volume 677, pages 1-27).
[0085] Compound half-life is inversely proportional to the
frequency of dosage of a compound. In vitro half-lifes of compounds
may be predicted from assays of microsomal half-life as described
by Kuhnz and Gieschen (Drug Metabolism and Disposition, (1998)
volume 26, pages 1120-1127).
[0086] The present invention also pertains to packaged
pharmaceutical compositions for treating disorders responsive to
CRF1 receptor modulation, e.g., treatment of anxiety, depression,
PTSD, cardiovascular disorder, headache and eating disorders by
CRF1 receptor modulation.
[0087] The packaged pharmaceutical compositions include a container
holding a therapeutically effective amount of at least one CRF1
receptor modulator as described supra and instructions (e.g.,
labeling) indicating the contained CRF1 receptor ligand is to be
used for treating a disorder responsive to CRF1 receptor modulation
in the patient.
[0088] Representative suitable synthetic routes for the preparation
of compounds of the invention are given in Schemes I and II. Those
having skill in the will recognize that the starting materials,
solvents, and reaction conditions may be varied and additional
steps employed to produce compounds encompassed by the present
invention. 17
[0089] The substituents Ar, R.sup.1, R.sup.2, R.sup.3, R.sup.4 and
R.sup.5 are as defined above for Formula I. 18
[0090] The definitions for the substituents A, Ar, R.sup.1,
R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are set forth above for
Formula I.
[0091] Compounds of the invention where R.sup.2 and R.sup.3
together represent
--(C.sub.0-C.sub.2)--G.sup.2--(C.sub.2-C.sub.4)-- may be prepared
by avoiding the aromatization portion (treatment with DDQ,
2,3-dichloro-5,6-dicyano-1,4-benzoquinone) of the synthesis set
forth in Scheme II.
[0092] The preparation of the compounds of the present invention is
illustrated further by the following examples, which are not to be
construed as limiting the invention in scope or spirit to the
specific procedures and compounds described in them.
[0093] The disclosures in this application of all articles and
references, including patents, are incorporated herein by
reference.
[0094] Commercial reagents were used without further purification.
THF refers to tetrahydrofuran. LDA refers to lithium
diisopropylamide. Room or ambient temperature refers to about 20 to
about 25.degree. C. Concentration means removal of solvent under
reduced pressure, e.g., by the use of a rotary evaporator. Mass
spectral data were obtained either by CI or APCI methods.
[0095] The following compounds are prepared essentially according
to the procedures set forth in Example 1, without the aromatization
procedure of Example 1B.
EXAMPLE 1
A. 2-Amino-4,5,6,7-tetrahydro-1-phenyl-1H-indole-3-carbonitrile
[0096] 19
[0097] A mixture of 2,4,6-trimethylaniline (500 g) and adipoin
(2-hydroxycyclohexanone, 464 g) in toluene (2.5 L) is heated to
reflux. A theoretical amount of water is removed azeotropically
over the course of 3 hours. The mixture is cooled to ambient
temperature, then malononitrile (244 g) and ammonium acetate (57 g)
are added. The reaction is slowly reheated back to reflux for about
1 hour with azeotropic removal of water. After cooling, the
precipitate that forms overnight is collected by filtration. The
dark solid is washed with ethanol and dried to afford 540 g of a
white powder: MS 280 (M+H).
B. 2-Amino-1-phenyl-1H-indole-3-carbonitrile
[0098] 20
[0099] Compound from Example 1A (20 g) is dissolved in 1,4-dioxane
(300 mL) and added DDQ (34 g) portionwise to the solution. The
reaction is stirred for 1 hour then filtered through celite to
remove insoluble side products. The filtrate is concentrated and
allowed to solidify. The product is collected by filtration and
washed with ethanol to yield 16 g of a tan colored powder: MS 276
(M+H).
C.
4-Hydroxy-2-methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole
[0100] 21
[0101] A mixture of compound from Example 1B (30 g), acetic
anhydride (15 mL) and acetic acid (30 mL) is refluxed for 1 hour
then concentrated to a solid. Phosphoric acid (40 mL, 85%) is added
to the amide. The mixture is then refluxed for 0.5 hours and cooled
to ambient temperature. The solution is poured onto ice and the
precipitate that forms is collected by filtration. The solids are
then washed with water and ethanol: MS 318 (M+H).
D.
4-Chloro-2-methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole
[0102] 22
[0103] A sample of the compound from Example 1B (2.2 g) is refluxed
in phosphoryl chloride (30 mL) for 3 hours. The excess phosphoryl
chloride is removed under reduced pressure and the residue
partitioned between aqueous potassium carbonate and
dichloromethane. The aqueous layer is extracted with a second
portion of dichloromethane. The combined extracts are then dried
over sodium sulfate, filtered and concentrated to give a tan
colored solid: MS 336 (M+H).
E.
4-(N,N-Dipropyl)amino-2-methyl-9-(2,4,6-trimethylphenyl)-9H-pyrimidino[-
4,5-b]indole (Compound 1)
[0104] 23
[0105] A mixture containing a sample of the compound prepared as
set forth above in Example 1D (1.0 g) and dipropylamine (1.0 mL) in
N-methylpyrrolidinone (5 mL) is heated to 100.degree. C. for 2
hours. The mixture is diluted with ethyl acetate and washed with
water, aqueous ammonium chloride, aqueous sodium bicarbonate, and
brine. The mixture is then dried over sodium sulfate, filtered and
concentrated. Purificarbon by flash chromatography using 10% ethyl
acetate in hexanes as eluant affords the title compound: MS 401
(M+H).
[0106] The following compounds are prepared essentially according
to the procedures set forth above in Example 1.
EXAMPLE 2
[0107] a)
4-(N-Cyclopropylmethyl-N-propyl)amino-2-methyl-9-(2,4,6-trimethy-
lphenyl)-9H-pyrimidino[4,5-b]indole: MS 413 (M+H).
[0108] b)
4-(N,N-Dipropyl)amino-2-methyl-9-(4-bromo-2,6-dimethylphenyl)-9H-
-pyrimidino[4,5-b]indole: MS 465 (M+H).
[0109] c)
4-(N,N-Dipropyl)amino-2-methyl-9-(2,6-dimethylphenyl)-9H-pyrimid-
ino [4,5-b]indole: MS 387 (M+H).
[0110] d)
4-Bis(2-methoxyethyl)amino-2-methyl-9-(4-bromo-2,6-dimethylpheny-
l)-9H-pyrimidino[4,5-b]indole: MS 497 (M+H).
[0111] e)
4-(N,N-Dipropyl)amino-2-methyl-9-(4-chloro-2-methylphenyl)-9H-py-
rimidino [4,5-b]indole: MS 407 (M+H).
[0112] f)
4-(N,N-Dipropyl)amino-2-methyl-9-(4-cyano-2,6-dimethylphenyl)-9H-
-pyrimidino[4,5-b]indole: MS 412 (M+H).
[0113] g)
4-(N,N-Dipropyl)amino-2-methyl-9-(4-aminomethyl-2,6-dimethylphen-
yl)-9H-pyrimidino[4,5-b]indole: MS 416 (M+H).
[0114] h)
4-(N,N-Dipropyl)amino-2-methyl-9-(2,6-dimethyl-4-(methylaminomet-
hyl)phenyl)-9H-pyrimidino [4,5-b]indole: MS 430 (M+H).
[0115] i)
4-Bis(2-methoxyethyl)amino-2-methyl-9-(2,4,6-trimethylphenyl)-9H-
-pyrimidino [4,5-b]indole: MS 433 (M+H) (Compound 3).
[0116] j) 4-(N-(2-Hydroxyethyl)
-N-Cyclopropylmethyl)amino-2-methyl-9-(2,4-
,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole: MS 415 (M+H).
[0117] k)
4-(N-(2-Hydroxyethyl)-N-Cyclopropyl)amino-2-methyl-9-(2,4,6-trim-
ethylphenyl)-9H-pyrimidino [4,5-b]indole: MS 401 (M+H).
[0118] l)
4-(N-Cyclopropylmethylamino)-2-methyl-9-(2,4,6-trimethylphenyl)--
9H-pyrimidino[4,5-b]indole: MS 371 (M+H).
EXAMPLE 3
[0119] The following compounds are prepared essentially according
to the procedures set forth in Example 1 but without the
aromatization procedure of Example 1B.
[0120] a)
4-(N-Cyclopropylmethyl-N-propyl)amino-9-(2,4,6-trimethylphenyl)--
5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 403 (M+H).
[0121] b)
4-(N,N-Dipropyl)amino-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrahyd-
ro-9H-pyrimidino[4,5-b]indole: MS 391 (M+H) (Compound 2).
[0122] c)
4-(N-Cyclopropylmethyl-N-propyl)amino-2-methyl-9-(2,4,6-trimethy-
lphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 417
(M+H).
[0123] d)
4-(N,N-Dipropyl)amino-2-methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8-
-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 405 (M+H).
[0124] e)
4-(N-Propyl-N-methyl)amino-2-methyl-9-(2,4,6-trimethylphenyl)-5,-
6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 377 (M+H).
[0125] f)
4-(N-Butyl-N-methyl)amino-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetr-
ahydro-9H-pyrimidino[4,5-b]indole: MS 377 (M+H).
[0126] g)
4-(N-Propyl)amino-8-(2,4,6-trimethylphenyl)-5,6,7-trihydrocyclop-
enta[2,1-d]-8H-pyrimidino[4,5-b]indole: MS 335 (M+H).
[0127] h)
4-(N,N-Dipropyl)amino-2-methyl-8-(2,4,6-trimethylphenyl)-5,6,7-t-
rihydrocyclopenta[2,1-d]-8H-pyrimidino[4,5-b]indole: MS 391 (M+H)
(Compound 6).
[0128] i)
4-Bis(2-methoxyethyl)amino-2-methyl-9-(2,4,6-trimethylphenyl)-5,-
6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 437 (M+H).
[0129] j) Ethyl
4-(dipropylamino)-9-(2,4,6-trimethylphenyl)-5,6,7,8-tetrah-
ydropyrimidino[4,5-b]indole-2-carboxylate: MS 463 (M+H).
[0130] k)
4-(N,N-Dipropyl)amino-2-carboxyamido-9-(2,4,6-trimethylphenyl)-5-
,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 434 (M+H).
[0131] l)
4-(N,N-Dipropyl)amino-2-aminomethyl-9-(2,4,6-trimethylphenyl)-5,-
6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 420 (M+H).
[0132] m)
4-(N,N-Dipropyl)amino-8-(2,4,6-trimethylphenyl)-5,6,7-trihydrocy-
clopenta[2,1-d]-8H-pyrimidino[4,5-b]indole: MS 377 (M+H).
[0133] n)
4-(N,N-Dibutylamino)-2-methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8--
tetrahydro-9H-pyrimidino[4,5-b]indole: MS 433 (M+H).
[0134] o)
4-(N,N-Dibutylamino)-2-methyl-9-(2,4,6-trimethylphenyl)-5,6,7,8--
tetrahydro-9H-pyrimidino[4,5-b]indole: MS 419 (M+H).
[0135] p)
4-(N-(2-(4-Chlorophenyl)ethyl)amino)-2-methyl-9-(2,4,6-trimethyl-
phenyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 459
(M+H).
[0136] q)
4-(N-(2-(4-Chlorophenyl)ethyl)amino)-2-methyl-9-(2,4-dimethylphe-
nyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 445
(M+H).
[0137] r)
4-(N-(4-Chlorophenyl)methylamino)-2-methyl-9-(2,4,6-trimethylphe-
nyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 445
(M+H).
[0138] s)
4-(N-(2-Methylphenyl)methylamino)-2-methyl-9-(2,4,6-trimethylphe-
nyl)-5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole: MS 425
(M+H).
[0139] t)
4-(N-(3-Trifluromethylphenyl)methylamino)-2-methyl-9-(2,4,6-trim-
ethylphenyl)-5,6,7,8-tetrahydro-9H-pyrimidino-[4,5-b]indole: MS 479
(M+H).
[0140] u)
4-(N-Propyl-N-methyl)amino-2-methyl-9-(2,4,6-trimethylphenyl)
5,6,7,8-tetrahydro-9H-pyrimidino[4,5-b]indole.
EXAMPLE 4
4-(N,N-Dipropyl)amino-6-ethylamino-2-methyl-9-(2,4,6-trimethylphenyl)-9H-p-
yrimidino[4,5-b]indole (Compound 5)
[0141] 24
[0142] A sample of the compound prepared in Example 1C (1.0 g) is
dissolved in a 30:1 mixture of acetic acid and 70% nitric acid (35
mL). The resulting solution is refuxed for 1.5 hours, cooled and
poured into cold water after which the solids are collected by
filtration and dried. The subsequent chlorination and substitution
reactions with phoshorous oxychloride and dipropylamine,
respectively are carried out according to Examples 1-D and 1-E. The
nitro functionality is then reduced by hydrogenation at 50 psi in a
solution of methanol (20 mL) containing 10% palladium on carbon
(0.5 g). After 6 hours the solution is filtered and concentrated.
Next, the aniline product is acylated by refluxing in acetic acid
(15 mL) for 6 hours. The solution is concentrated and extracted
from aqueous sodium carbonate solution with dichloromethane. The
extract is then dried over sodium sulfate, filtered and
concentrated. The resulting amide is dissolved in tetrahydrofuran
(15 mL) and treated with borane-methylsulfide complex (0.6 mL). The
solution is refluxed for 3 hours, quenched with a large excess of
methanol (20 mL) and concentrated. The final product is purified by
flash chromatography using 10% ethyl acetate in hexanes as eluant:
MS 444 (M+H).
EXAMPLE 5
[0143] The following compounds are prepared essentially according
to the procedures set forth above in Example 4.
[0144] a)
4-(N,N-Dipropyl)amino-6-diethylamino-2-methyl-9-(2,4,6-trimethyl-
phenyl)-9H-pyrimidino[4,5-b]indole: MS 472 (M+H).
[0145] b)
4-(N-Cyclopropylmethyl-N-propyl)amino-6-ethylamino-2-methyl-9-(2-
,4,6-trimethylphenyl)-9H-pyrimidino[4,5-b]indole: MS 456 (M+H).
[0146] c)
4-(N,N-dipropylamino)-2-methyl-6dimethylamino-9-(2,4,6-trimethyl-
phenyl)-9H-pyrimidino[4,5-b]indole: MS 444 (M+H).
[0147] The following two assays for human CRF1 receptor activity
are standard assays of CRF binding that may be used to determine
the affinity of CRF for the CRF receptor. The second assay,
described in Example 7 is also a CRF1 signal transduction
assay.
EXAMPLE 6
Assay for Recombinant Human CRF.sub.1 Receptor Binding Activity
[0148] CRF receptor binding is performed using a modified version
of the assay described by Grigoriadis and De Souza (Methods in
Neurosciences, Vol. 5, 1991). Membrane pellets containing CRF
receptors are re-suspended in 50 mM Tris buffer pH 7.7 containing
10 mM MgCl.sub.2 and 2 mM EDTA and centrifuged for 10 minutes at
48000 g. Membranes are washed again and brought to a final
concentration of 1500 mg/ml in binding buffer (Tris buffer above
with 0.1% BSA, 15 mM bacitracin and 0.01 mg/mL aprotinin). For the
binding assay, 100 mL of the membrane preparation is added to 96
well microtube plates containing 100 mL of .sup.125I-CRF (SA 2200
Ci/mmol, final concentration of 100 pM) and 50 mL of drug. Binding
is carried out at room temperature for 2 hours. Plates are then
harvested on a Brandel 96 well cell harvester and filters are
counted for gamma emissions on a Wallac 1205 Betaplate liquid
scintillation counter. Non specific binding is defined by 1 mM cold
CRF. IC.sub.50 values are calculated with the non-linear curve
fitting program RS/1 (BBN Software Products Corp., Cambridge,
Mass.). The binding affinity for the compounds of Formula I
expressed as IC.sub.50 value, generally ranges from about 0.5
nanomolar to about 10 micromolar.
[0149] Preferred compounds of the invention exhibit good activity
in standard in vitro receptor binding assays, specifically the
assay set forth in Example 7 below. Particularly preferred
compounds of the invention have an IC.sub.50 of about 10 micromolar
or less, still more preferably an IC.sub.50 of about 100 nanomolar
or less, even more preferably an IC.sub.50 of about 10 nanomolar or
less, or even 1 nanomolar or less in such a defined standard in
vitro CRF receptor binding assay.
EXAMPLE 7
Assay for Human CRF Receptor Binding Activity in IMR32 Cells
[0150] Alternatively, the binding activity of the compounds of
formula I to the human CRF.sub.1 receptor can be measured as
follows:
[0151] IMR-32 human neuroblastoma cells are grown to 80% confluence
in EMEM containing Earle's Balanced Salts and 2 mM l-glutamine with
10% FBS, 25 mM HEPES, 1 mM Sodium Pyruvate, and nonessential amino
acids. At this time, flasks of cells are treated with 2.5 uM
5-bromo-2'-deoxyuridine (Br-dU) for 10 days. Media is changed every
3-4 days across the 10-day period. Cells are harvested using
No-Zyme (JRH Biosciences) and rinsed with PBS. For membrane
preparation, cells are homogenized in wash buffer (50 mM Tris HCl,
10 mM MgCl.sub.2, 2 mM EGTA, pH 7.4) and centrifuged at
48,000.times.g for 10 minutes at 4.degree. C. Pellets are
resuspended, homogenized and centrifuged two additional times. The
receptor binding assay is performed using assay buffer (50 mM Tris
HCl, 10 mM MgCl.sub.2, 2 mM EGTA, pH 7.4, 0.1% BSA, 0.1 mM
bacitracin (22.0 mg/100 mL)), 150 ug protein/tube, and
[.sup.125I]Sauvagine (NEN; 100 pM for competition analysis and 10
pM-1 nM for saturation analysis) to yield a final volume of 200
.mu.L. Nonspecific binding is defined using 2 .mu.M r/h CRF or 9-41
alpha-helical CRF. Cells are incubated for 2 hours at room
temperature. The assay is terminated by rapid vacuum filtration
(Tomtec: Deepwell 3) through GFC filters presoaked in 1% PEI using
ice-cold 50 mM Tris Hcl and dry thoroughly by air. Specific
Binding: 70-80%; Kd (nM): 0.30 nM; Bmax (fmole/mg protein): 40-50.
IC.sub.50 values are calculated with the non-linear curve fitting
program RS/1 (BBN Software Products Corp., Cambridge, Mass.).
EXAMPLE 8
Preparation of Radiolabeled Probe Compounds of the Invention
[0152] The compounds of the invention are prepared as radiolabeled
probes by carrying out their synthesis using precursors comprising
at least one atom that is a radioisotope. The radioisotope is
preferably selected from of at least one of carbon (preferably
.sup.14C), hydrogen (preferably .sup.3H), sulfur (preferably
.sup.35S), or iodine (preferably .sup.125I). Such radiolabeled
probes are conveniently synthesized by a radioisotope supplier
specializing in custom synthesis of radiolabeled probe compounds.
Such suppliers include Amersham Corporation, Arlington Heights,
Ill.; Cambridge Isotope Laboratories, Inc. Andover, Mass.; SRI
International, Menlo Park, Calif.; Wizard Laboratories, West
Sacramento, Calif.; ChemSyn Laboratories, Lexena, KS; American
Radiolabeled Chemicals, Inc., St. Louis, Mo.; and Moravek
Biochemicals Inc., Brea, Calif.
[0153] Tritium labeled probe compounds are also conveniently
prepared catalytically via platinum-catalyzed exchange in tritiated
acetic acid, acid-catalyzed exchange in tritiated trifluoroacetic
acid, or heterogeneous-catalyzed exchange with tritium gas. Such
preparations are also conveniently carried out as a custom
radiolabeling by any of the suppliers listed in the preceding
paragraph using the compound of the invention as substrate. In
addition, certain precursors may be subjected to tritium-halogen
exchange with tritium gas, tritium gas reduction of unsaturated
bonds, or reduction using sodium borotritide, as appropriate.
EXAMPLE 9
Receptor Autoradiography
[0154] Receptor autoradiography (receptor mapping) is carried out
in vitro as described by Kuhar in sections 8.1.1 to 8.1.9 of
Current Protocols in Pharmacology (1998) John Wiley & Sons, New
York, using radiolabeled compounds of the invention prepared as
described in the preceding Example.
[0155] The invention and the manner and process of making and using
it, are now described in such full, clear, concise and exact terms
as to enable any person skilled in the art to which it pertains, to
make and use the same. It is to be understood that the foregoing
describes preferred embodiments of the present invention and that
modifications may be made therein without departing from the spirit
or scope of the present invention as set forth in the claims. To
particularly point out and distinctly claim the subject matter
regarded as invention, the following claims conclude this
specification.
* * * * *