U.S. patent application number 10/092312 was filed with the patent office on 2003-07-03 for 4 - (2-butylamino) - 2, 7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) pyrazolo- [1,5-a] - 1,3,5-triazine, its enantiomers and pharmaceutically acceptable salts as corticotropin releasing factor receptor ligands.
Invention is credited to Gilligan, Paul J..
Application Number | 20030125330 10/092312 |
Document ID | / |
Family ID | 23052141 |
Filed Date | 2003-07-03 |
United States Patent
Application |
20030125330 |
Kind Code |
A1 |
Gilligan, Paul J. |
July 3, 2003 |
4 - (2-butylamino) - 2, 7-dimethyl-8-
(2-methyl-6-methoxypyrid-3-yl) pyrazolo- [1,5-A] - 1,3,5-triazine,
its enantiomers and pharmaceutically acceptable salts as
corticotropin releasing factor receptor ligands
Abstract
Corticotropin releasing factor (CRF) antagonists of Formula (I):
1 and its use in treating anxiety, depression, and other
psychiatric, neurological disorders as well as treatment of
immunological, cardiovascular or heart-related diseases and colonic
hypersensitivity associated with psychopathological disturbance and
stress.
Inventors: |
Gilligan, Paul J.;
(US) |
Correspondence
Address: |
STEPHEN B. DAVIS
BRISTOL-MYERS SQUIBB COMPANY
PATENT DEPARTMENT
P O BOX 4000
PRINCETON
NJ
08543-4000
US
|
Family ID: |
23052141 |
Appl. No.: |
10/092312 |
Filed: |
March 6, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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60275403 |
Mar 13, 2001 |
|
|
|
Current U.S.
Class: |
514/246 ;
544/180 |
Current CPC
Class: |
A61P 3/04 20180101; A61P
31/18 20180101; A61P 3/08 20180101; A61P 25/24 20180101; A61P 25/00
20180101; A61P 25/32 20180101; A61P 11/00 20180101; A61P 25/04
20180101; A61P 37/06 20180101; A61P 29/00 20180101; A61P 9/00
20180101; A61K 31/53 20130101; A61P 25/06 20180101; A61P 35/00
20180101; A61P 43/00 20180101; A61P 25/02 20180101; A61P 19/02
20180101; A61P 25/16 20180101; A61P 25/08 20180101; A61P 1/00
20180101; A61P 17/00 20180101; A61P 25/22 20180101; A61P 19/00
20180101; C07D 487/04 20130101; A61P 17/06 20180101; A61P 25/30
20180101; A61P 15/00 20180101; A61P 25/28 20180101; A61P 1/14
20180101; A61P 21/00 20180101; A61P 19/10 20180101; A61P 1/04
20180101; A61P 11/06 20180101; A61P 15/08 20180101; A61P 25/12
20180101; A61P 9/12 20180101 |
Class at
Publication: |
514/246 ;
544/180 |
International
Class: |
A61K 031/53; C07D
487/04 |
Claims
What is claimed is:
1. A compound of Formula (I): 4isomers thereof, stereoisomeric
forms thereof, or mixtures of stereoisomeric forms thereof,
pharmaceutically acceptable prodrugs thereof, or pharmaceutically
acceptable salt forms.
2. A compound of claim 1, pharmaceutically acceptable prodrugs
thereof, or pharmaceutically acceptable salt forms thereof, wherein
said compound is
4-((R)-2-butylamino)2,7-dimethyl-8-(2-methyl-6-methoxypyrid-3-yl)
[1,5-a]-pyrazolo-1,3,5-triazine.
3. A compound of claim 1, pharmaceutically acceptable prodrugs
thereof, or pharmaceutically acceptable salt forms thereof, wherein
said compound is substantially free of its (S) stereoisomer.
4. A compound of claim 1, wherein said compound is
4-(2-butylamino)2,7-dim- ethyl-8-(2-methyl-6-methoxypyrid-3-yl)
[1,5-a]-pyrazolo-1,3,5-triazine.
5. A compound of claim 1, wherein said compound is
4-((R)-2-butylamino)2,7- -dimethyl-8-(2-methyl-6-methoxypyrid-3-yl)
[1,5-a]-pyrazolo-1,3,5-triazine- .
6. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and a therapeutically effective amount of a
compound of claim 1.
7. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and a therapeutically effective amount of a
compound of claim 5.
8. A method of antagonizing a CRF receptor in a mammal, comprising
administering to the mammal, a therapeutically effective amount of
a compound as claimed in claim 1.
9. A method of treating a disorder manifesting hypersecretion of
CRF in a warm-blooded animal, comprising administering to the
animal a therapeutically effective amount of a compound as claimed
in claim 1.
10. A method for the treatment of a disorder, the treatment of
which can be effected or facilitated by antagonizing CRF,
comprising administering to the mammal a therapeutically effective
amount of a compound of claim 1.
11. A method of antagonizing a CRF receptor in a mammal, comprising
administering to the mammal, a therapeutically effective amount of
a compound as claimed in claim 5.
12. A method of treating anxiety or depression in mammals,
comprising administering to the mammal a therapeutically effective
amount of a compound of claim 1.
13. A method of treating anxiety or depression in mammals,
comprising administering to the mammal a therapeutically effective
amount of a compound of claim 5.
14. A method for screening for ligands for CRF receptors, which
method comprises: a) carrying out a competitive binding assay with
a CRF receptor, a compound according to claim 1 which is labelled
with a detectable label, and a candidate ligand; and b) determining
the ability of said candidate ligand to displace said labelled
compound.
15. A method for detecting CRF receptors in tissue comprising: a)
contacting a compound of claim 1, which is labelled with a
detectable label, with a tissue, under conditions that permit
binding of the compound to the tissue; and b) detecting the
labelled compound bound to the tissue.
16. A method of inhibiting the binding of CRF to a CRF-1 receptor,
comprising contacting a compound of claim 1 with a solution
comprising cells expressing the CRF1 receptor, wherein the compound
is present in the solution at a concentration sufficient to inhibit
the binding of CRF to the CRF-1 receptor.
17. An article of manufacture comprising: a) a packaging material;
b) a compound of claim 1; and c) a label or package insert
contained within said packaging material idicating that said
compound is effective for treating anxiety or depression.
Description
FIELD OF THE INVENTION
[0001] This invention relates to a treatment of psychiatric
disorders and neurological diseases including major depression,
anxiety-related disorders, post-traumatic stress disorder,
supranuclear palsy and feeding disorders as well as treatment of
immunological, cardiovascular or heart-related diseases and colonic
hypersensitivity associated with psycho-pathological disturbances
and stress, by administration of
4-(2-Butylamino)-2,7-dimethyl-8-(2-methyl-6-methoxypyrid-3-yl)pyrazolo-[1-
,5-a]-1,3,5-triazine, its enantiomer and pharmaceutically
acceptable salts as a corticotropin releasing factor receptor
ligand.
BACKGROUND OF THE INVENTION
[0002] Corticotropin releasing factor (herein referred to as CRF),
a 41 amino acid peptide, is the primary physiological regulator of
proopiomelanocortin (POMC)--derived peptide secretion from the
anterior pituitary gland [J. Rivier et al., Proc. Nat. Acad. Sci.
(USA 80:4851 (1983); W. Vale et al., Science 213:1394 (1981)]. 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 [W. Vale et al.,
Rec. Prog. Horm. Res. 39:245 (1983); G. F. Koob, Persp. Behav. Med.
2:39 (1985); E. B. De Souza et al., J. Neurosci. 5:3189 (1985)].
There is also evidence that CRF plays a significant role in
integrating the response of the immune system to physiological,
psychological, and immunological stressors [J. E. Blalock,
Physiological Reviews 69:1 (1989); J. E. Morley, Life Sci. 41:527
(1987)].
[0003] 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 [for review see E. B. De
Souza, Hosp. Practice 23:59 (1988)].
[0004] In affective disorder, or major depression, the
concentration of CRF is significantly increased in the
cerebrospinal fluid (CSF) of drug-free individuals [C. B. Nemeroff
et al., Science 226:1342 (1984); C. M. Banki et al., Am. J.
Psychiatry 144:873 (1987); R. D. France et al., Biol. Psychiatry
28:86 (1988); M. Arato et al., Biol Psychiatry 25:355 (1989)].
Furthermore, the density of CRF receptors is significantly
decreased in the frontal cortex of suicide victims, consistent with
a hypersecretion of CRF [C. B. Nemeroff et al., Arch. Gen.
Psychiatry 45:577 (1988)]. In addition, there is a blunted
adrenocorticotropin (ACTH) response to CRF (i.v. administered)
observed in depressed patients [P. W. Gold et al., Am J. Psychiatry
141:619 (1984) F. Holsboer et al., Psychoneuroendocrinology 9:147
(1984); P. W. Gold et al., New Eng. J. Med. 314:1129 (1986)].
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 [R. M.
Sapolsky, Arch. Gen. Psychiatry 46:1047 (1989)]. There is
preliminary evidence that tricyclic antidepressants can alter CRF
levels and thus modulate the numbers of CRF receptors in brain
[Grigoriadis et al., Neuropsychopharmacology 2:53 (1989)].
[0005] It has also been postulated that CRF has a role 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 [D. R.
Britton et al., Life Sci. 31:363 (1982); C. W. Berridge and A. J.
Dunn Regul. Peptides 16:83 (1986)]. Preliminary studies using the
putative CRF receptor antagonist a-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 [C. W. Berridge and A. J. Dunn Horm. Behav.
21:393 (1987), Brain Research Reviews 15:71 (1990)].
[0006] 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 [K. T. Britton et al.,
Psychopharmacology 86:170 (1985); K. T. Britton et al.,
Psychopharmacology 94:306 (1988)] and in the acoustic startle test
[N. R. Swerdlow et al., Psychopharmacology 88:147 (1986)] in rats.
The benzodiazepine receptor antagonist (Ro15-1788), 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 (FG7142) enhanced the actions of CRF
[K. T. Britton et al., Psychopharmacology 94:306 (1988)].
[0007] The mechanisms and sites of action through which the
standard anxiolytics and antidepressants produce their therapeutic
effects remain to be elucidated. It has been hypothesized however,
that they are involved in the suppression of the CRF hypersecretion
that is observed in these disorders. Of particular interest is that
preliminary studies examining the effects of a CRF receptor
antagonist (a-helical CRF9-41) in a variety of behavioral paradigms
have demonstrated that the CRF antagonist produces
"anxiolytic-like" effects qualitatively similar to the
benzodiazepines [for review see G.F. Koob and K. T. Britton, In:
Corticotropin-Releasing Factor: Basic and Clinical Studies of a
Neuropeptide, E. B. De Souza and C. B. Nemeroff eds., CRC Press
p221 (1990)].
[0008] It has been further postulated that CRF has a role in
cardiovascular or heart-related diseases as well as
gastrointestinal disorders arising from stress such as
hypertension, tachycardia and congestive heart failure, stroke,
irritable bowel syndrome post-operative ileus and colonic
hypersensitivity associated with psychopathological disturbance and
stress [for reviews see E. D. DeSouza, C. B. Nemeroff, Editors;
Corticotropin-Releasing Factor: Basic and Clinical Studies of a
Neuropeptide, E. B. De Souza and C. B. Nemeroff eds., CRC Press
p221 (1990) and C. Maillot, M. Million, J. Y. Wei, A. Gauthier, Y.
Tache, Gastroenterology, 119, 1569-1579 (2000)].
[0009] Over-expression or under-expression of CRF has been proposed
as an underlying cause for several medical disorders. Such
treatable disorders include, for example and without limitation:
affective disorder, anxiety, depression, headache, irritable bowel
syndrome, post-traumatic stress disorder, supranuclear palsy,
immune suppression, Alzheimer's disease, gastrointestinal diseases,
anorexia nervosa or other feeding disorder, drug addiction, drug or
alcohol withdrawal symptoms, inflammatory diseases, cardiovascular
or heart-related diseases, fertility problems, human
immunodeficiency virus infections, hemorrhagic stress, obesity,
infertility, head and spinal cord traumas, epilepsy, stroke,
ulcers, amyotrophic lateral sclerosis, hypoglycemia, hypertension,
tachycardia and congestive heart failure, stroke, osteoporosis,
premature birth, psychosocial dwarfism, stress-induced fever,
ulcer, diarrhea, post-operative ileus and colonic hypersensitivity
associated with psychopathological disturbance and stress [for
reviews see J. R. McCarthy, S. C. Heinrichs and D. E. Grigoriadis,
Cuur. Pharm. Res., 5, 289-315 (1999); P. J. Gilligan, D. W.
Robertson and R. Zaczek, J. Medicinal Chem., 43, 1641-1660 (2000),
G. P. Chrousos, Int. J. Obesity, 24, Suppl. 2, S50-S55 (2000); E.
Webster, D. J. Torpy, I. J. Elenkov, G. P. Chrousos, Ann. N.Y.
Acad. Sci., 840, 21-32 (1998); D. J. Newport and C. B. Nemeroff,
Curr. Opin. Neurobiology, 10, 211-218 (2000); G. Mastorakos and I.
Ilias, Ann. N.Y. Acad. Sci., 900, 95-106 (2000); M. J. Owens and C.
B. Nemeroff, Expert Opin. Invest. Drugs, 8, 1849-1858 (1999); G. F.
Koob, Ann. N.Y. Acad. Sci., 909, 170-185 (2000)].
[0010] The following publications each describe CRF antagonist
compounds; however, none disclose the compounds provided herein:
WO95/10506; WO99/51608; WO97/35539; WO99/01439; WO97/44308;
WO97/35846; WO98/03510; WO99/11643; PCT/US99/18707; WO99/01454;
and, WO00/01675.
SUMMARY OF THE INVENTION
[0011] In accordance with one aspect, the present invention
provides a novel compound, pharmaceutical compositions and methods
which may be used in the treatment of affective disorder, anxiety,
depression, irritable bowel syndrome, post-traumatic stress
disorder, supranuclear palsy, immune suppression, Alzheimer's
disease, gastrointestinal disease, anorexia nervosa or other
feeding disorder, drug or alcohol withdrawal symptoms, drug
addiction, inflammatory disorder, fertility problems, disorders,
the treatment of which can be effected or facilitated by
antagonizing CRF, including but not limited to disorders induced or
facilitated by CRF, or a disorder selected from inflammatory
disorders such as rheumatoid arthritis and osteoarthritis, pain,
asthma, psoriasis and allergies; generalized anxiety disorder;
panic, phobias, obsessive-compulsive disorder; post-traumatic
stress disorder; sleep disorders induced by stress; pain perception
such as fibromyalgia; mood disorders such as depression, including
major depression, single episode depression, recurrent depression,
child abuse induced depression, and postpartum depression;
dysthemia; bipolar disorders; cyclothymia; fatigue syndrome;
stress-induced headache; cancer, human immunodeficiency virus (HIV)
infections; neurodegenerative diseases such as Alzheimer's disease,
Parkinson's disease and Huntington's disease; gastrointestinal
diseases such as ulcers, irritable bowel syndrome, Crohn's disease,
spastic colon, diarrhea, and post operative ilius and colonic
hypersensitivity associated by psychopathological disturbances or
stress; eating disorders such as anorexia and bulimia nervosa;
hemorrhagic stress; stress-induced psychotic episodes; euthyroid
sick syndrome; syndrome of inappropriate antidiarrhetic hormone
(ADH); obesity; infertility; head traumas; spinal cord trauma;
ischemic neuronal damage (e.g., cerebral ischemia such as cerebral
hippocampal ischemia); excitotoxic neuronal damage; epilepsy;
cardiovascular and hear related disorders including hypertension,
tachycardia and congestive heart failure; stroke; immune
dysfunctions including stress induced immune dysfunctions (e.g.,
stress induced fevers, porcine stress syndrome, bovine shipping
fever, equine paroxysmal fibrillation, and dysfunctions induced by
confinement in chickens, sheering stress in sheep or human-animal
interaction related stress in dogs); muscular spasms; urinary
incontinence; senile dementia of the Alzheimer's type; multiinfarct
dementia; amyotrophic lateral sclerosis; chemical dependencies and
addictions (e.g., dependencies on alcohol, cocaine, heroin,
benzodiazepines, or other drugs); drug and alcohol withdrawal
symptoms; osteoporosis; psychosocial dwarfism; and hypoglycemia in
a mammal.
[0012] The present invention provides a novel compound that binds
to corticotropin releasing factor receptors, thereby altering the
anxiogenic effects of CRF secretion. The compound of the present
invention is useful for the treatment of psychiatric disorders and
neurological diseases, anxiety-related disorders, post-traumatic
stress disorder, 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 in a mammal.
[0013] According to another aspect, the present invention provides
a novel compound of Formula (I) (described below) which is useful
as an antagonist of the corticotropin releasing factor. The
compound of the present invention exhibits activity as a
corticotropin releasing factor antagonist and appears to suppress
CRF hypersecretion. The present invention also includes
pharmaceutical compositions containing such a compound of Formula
(I), and methods of using such a compound for the suppression of
CRF hypersecretion, and/or for the treatment of anxiogenic
disorders.
[0014] The use of competitive binding assays is considered
particularly valuable for screening candidates for new drugs, e.g.
to identify new CRF ligands or other compounds having even greater
or more selective binding affinity for CRF receptors, which
candidates would therefore be potentially useful as drugs. In the
assay, one determines the ability of the candidate ligand to
displace the labelled compound.
[0015] Therefore, another embodiment of the invention includes the
use of a compound of the invention is a binding assay, wherein one
or more of the compounds may be joined to a label, where the label
can directly or indirectly provide a detectable signal. Various
labels include radioisotopes, fluorescers, chemiluminescers,
specific binding molecules, particles, e.g. magnetic particles, and
the like.
[0016] Another embodiment of the invention is directed 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.
DETAILED DESCRIPTION OF THE INVENTION
[0017] [1] In a first embodiment, the present invention provides a
compound of Formula (I): 2
[0018] and stereoisomeric forms thereof, or mixtures of
stereoisomeric forms thereof, and pharmaceutically acceptable salt
or pro-drug forms thereof.
[0019] [2] In another embodiment, the present invention provides a
compound of embodiment [1], isomers thereof, stereoisomeric forms
thereof, mixtures of stereoisomeric forms thereof, pharmaceutically
acceptable prodrugs thereof, or pharmaceutically acceptable salt
forms thereof, wherein said compound is
4-((R)-2-butylamino)2,7-dimethyl-8-(2-m- ethyl-6-methoxypyrid-3-yl)
[1,5-a]-pyrazolo-1,3,5-triazine.
[0020] [3] In another embodiment, the present invention provides a
compound of any one of embodiments [1] to [2], pharmaceutically
acceptable prodrugs thereof, or pharmaceutically acceptable salt
forms thereof, wherein said compound is substantially free of its
(S) stereoisomer
[0021] [4] In another embodiment, the present invention provides a
compound of embodiment [1], wherein said compound is
4-(2-butylamino)2,7-dimethyl-8-(2-methyl-6-methoxypyrid-3-yl)
[1,5-a]-pyrazolo-1,3,5-triazine.
[0022] [5] In another embodiment, the present invention provides a
compound of embodiment [1], wherein said compound is
4-((R)-2-butylamino)2,7-dimethyl-8-(2-methyl-6-methoxypyrid-3-yl)
[1,5-a]-pyrazolo-1,3,5-triazine.
[0023] [6] A pharmaceutical composition comprising a
pharmaceutically acceptable carrier and a therapeutically effective
amount of a compound of any one of embodiments [1] to [5].
[0024] [7] In another embodiment, the present invention provides a
method of antagonizing a CRF receptor in a mammal, comprising
administering to the mammal, a therapeutically effective amount of
a compound of any one of embodiments [1] to [5].
[0025] [8] In another embodiment, the present invention provides a
method of treating a disorder manifesting hypersecretion of CRF in
a warm-blooded animal, comprising administering to the animal a
therapeutically effective amount of a compound of any one of
embodiments [1] to [5].
[0026] [9] In another embodiment, the present invention provides a
method for the treatment of a disorder, the treatment of which can
be effected or facilitated by antagonizing CRF, comprising
administering to the mammal a therapeutically effective of a
compound of any one of embodiments [1] to [5].
[0027] [10] In another embodiment, the present invention provides a
method of antagonizing a CRF receptor in a mammal, comprising
administering to the mammal, a therapeutically effective amount of
a compound of any one of embodiments [1] to [5].
[0028] [11] In another embodiment, the present invention provides a
method of treating anxiety or depression in mammals, comprising
administering to the mammal a therapeutically effective amount of a
compound of any one of embodiments [1] to [5].
[0029] [12] In another embodiment, the present invention provides a
method for screening for ligands for CRF receptors, which method
comprises:
[0030] a) carrying out a competitive binding assay with a CRF
receptor, a compound of any one of embodiments [1] to [5] which is
labelled with a detectable label, and a candidate ligand; and
[0031] b) determining the ability of said candidate ligand to
displace said labelled compound.
[0032] [13] In another embodiment, the present invention provides a
method for detecting CRF receptors in tissue comprising:
[0033] a) contacting a compound of any one of embodiments [1] to
[5], which is labelled with a detectable label, with a tissue,
under conditions that permit binding of the compound to the tissue;
and
[0034] b) detecting the labelled compound bound to the tissue.
[0035] [14] In another embodiment, the present invention provides a
method of inhibiting the binding of CRF to a CRF-1 receptor,
comprising contacting a compound of any one of embodiments [1] to
[5] with a solution comprising cells expressing the CRF1 receptor,
wherein the compound is present in the solution at a concentration
sufficient to inhibit the binding of CRF to the CRF-1 receptor.
[0036] [15] In another embodiment, the present invention provides a
article of manufacture comprising:
[0037] a) a packaging material;
[0038] b) a compound of any one of embodiments [1] to [5] ; and
[0039] c) a label or package insert contained within said packaging
material idicating that said compound is effective for treating
anxiety or depression.
[0040] [16] The present invention also comprises a method of
treating affective disorder, anxiety, depression, headache,
irritable bowel syndrome, post-traumatic stress disorder,
supranuclear palsy, immune suppression, Alzheimer's disease,
gastrointestinal diseases, anorexia nervosa or other feeding
disorder, drug addiction, drug or alcohol withdrawal symptoms,
inflammatory diseases, cardiovascular or heart-related diseases,
fertility problems, human immunodeficiency virus infections,
hemorrhagic stress, obesity, infertility, head and spinal cord
traumas, epilepsy, stroke, ulcers, amyotrophic lateral sclerosis,
hypoglycemia or a disorder the treatment of which can be effected
or facilitated by antagonizing CRF, including but not limited to
disorders induced or facilitated by CRF, in mammals comprising
administering to the mammal a therapeutically effective amount of a
compound of any one of embodiments [1] to [5].
[0041] Definitions
[0042] As used herein, the term "pharmaceutically acceptable salts"
refers to salts prepared from pharmaceutically acceptable non-toxic
acids, including inorganic acids and organic acids. Suitable
non-toxic acids include inorganic and organic acids of basic
residues such as amines, for example, acetic, benzenesulfonic,
benzoic, amphorsulfonic, citric, ethenesulfonic, fumaric, gluconic,
glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic,
malic, mandelic, methanesulfonic, mucic, nitric, pamoic,
pantothenic, phosphoric, succinic, sulfuric, tartaric acid,
p-toluenesulfonic and the like; and alkali or organic salts of
acidic residues such as carboxylic acids, for example, alkali and
alkaline earth metal salts derived from the following bases: sodium
hydride, sodium hydroxide, potassium hydroxide, calcium hydroxide,
aluminum hydroxide, lithium hydroxide, magnesium hydroxide, zinc
hydroxide, ammonia, trimethylammonia, triethylammonia,
ethylenediamine, n-methyl-glucamine, lysine, arginine, ornithine,
choline, N,N'-dibenzylethylenediamine, chloroprocaine,
diethanolamine, procaine, n-benzylphenethylamine, diethylamine,
piperazine, tris(hydroxymethyl)-ami- nomethane, tetramethylammonium
hydroxide, and the like.
[0043] Pharmaceutically acceptable salts of the compounds of the
invention can be prepared by reacting the free acid or base forms
of these compounds with a stoichiometric amount of the appropriate
base or acid in water or in an organic solvent, or in a mixture of
the two; generally, nonaqueous media like ether, ethyl acetate,
ethanol, isopropanol, or acetonitrile are preferred. Lists of
suitable salts are found in Remington's Pharmaceutical Sciences,
17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, the
disclosure of which is hereby incorporated by reference.
[0044] "Pharmaceutically acceptable prodrugs" as used herein means
any covalently bonded carriers which release the active parent drug
of Formula (I) in vivo when such prodrug is administered to a
mammalian subject. Prodrugs of the compounds of Formula (I) are,
within the scope of sound medical judgment, suitable for use in
contact with the tissues of humans and lower animals with undue
toxicity, irritation, allergic response, and the like, commensurate
with a reasonable benefit/risk ratio, and effective for their
intended use, as well as the zwitterionic forms, where possible, of
the compounds of the invention. The term "prodrug" means compounds
that are rapidly transformed in vivo to yield the parent compound
of formula (I), for example by hydrolysis in blood. Functional
groups which may be rapidly transformed, by metabolic cleavage, in
vivo form a class of groups reactive with the carboxyl group of the
compounds of this invention. They include, but are not limited to
such groups as alkanoyl (such as acetyl, propionyl, butyryl, and
the like), unsubstituted and substituted aroyl (such as benzoyl and
substituted benzoyl), alkoxycarbonyl (such as ethoxycarbonyl),
trialkylsilyl (such as trimethyl- and triethysilyl) monoesters
formed with dicarboxylic acids (such as succinyl), and the like.
Because of the ease with which the metabolically cleavable groups
of the compounds useful according to this invention are cleaved in
vivo, the compounds bearing such groups act as pro-drugs. The
compounds bearing the metabolically cleavable groups have the
advantage that they may exhibit improved bioavailability as a
result of enhanced solubility and/or rate of absorption conferred
upon the parent compound by virtue of the presence of the
metabolically cleavable group. A thorough discussion of prodrugs is
provided in the following: Design of Prodrugs, H. Bundgaard, ed.,
Elsevier, 1985; Methods in Enzymology, K. Widder et al, Ed.,
Academic Press, 42, p. 309-396, 1985; A Textbook of Drug Design and
Development, Krogsgaard-Larsen and H. Bundgaard, ed., Chapter 5;
Design and Applications of Prodrugsp. 113-191, 1991; Advanced Drug
Delivery Reviews, H. Bundgard, 8, p. 1-38, 1992; Journal of
Pharmaceutical Sciences, 77, p. 285, 1988; Chem. Pharm. Bull., N.
Nakeya et al, 32, p. 692, 1984; Pro-drugs as Novel Delivery
Systems, T. Higuchi and V. Stella, Vol. 14 of the A.C.S. Symposium
Series, and Bioreversible Carriers in Drug Design, Edward B. Roche,
ed., American Pharmaceutical Association and Pergamon Press, 1987,
which are incorporated herein by reference.
[0045] "Prodrugs" are considered to be any covalently bonded
carriers which release the active parent drug of Formula (I) in
vivo when such prodrug is administered to a mammalian subject.
Prodrugs of the compounds of Formula (I) are prepared by modifying
functional groups present in the compounds in such a way that the
modifications are cleaved, either in routine manipulation or in
vivo, to the parent compounds. Prodrugs include compounds wherein
hydroxy, amine, or sulfhydryl groups are bonded to any group that,
when administered to a mammalian subject, cleaves to form a free
hydroxyl, amino, or sulfhydryl group, respectively. Examples of
prodrugs include, but are not limited to, acetate, formate and
benzoate derivatives of alcohol and amine functional groups in the
compounds of Formula (I), and the like.
[0046] As used herein to describe a compound, the term
"substantially free of its (S) stereoisomer" means that the
compound is made up of a significantly greater proportion of its
(R) stereoisomer than of its optical antipode (i.e., its (S)
stereoisomer). In a preferred embodiment of the invention, the term
"substantially free of its (S) stereoisomer" means that the
compound is made up of at least about 90% by weight of its (R)
stereoisomer and about 10% by weight or less of its (S)
stereoisomer.
[0047] In a more preferred embodiment of the invention, the term
"substantially free of its (S) stereoisomer" means that the
compound is made up of at least about 95% by weight of its (R)
stereoisomer and about 5% by weight or less of its (S)
stereoisomer. In an even more preferred embodiment, the term
"substantially free of its (S) stereoisomer" means that the
compound is made up of at least about 99% by weight of its (R)
stereoisomer and about 1% or less of its (S) stereoisomer. In
another preferred embodiment, the term "substantially free of its
(S) stereoisomer" means that the compound is made up of nearly 100%
by weight of its (R) stereoisomer. The above percentages are based
on the total amount of the combined stereoisomers of the
compound.
[0048] The term "therapeutically effective amount" of a compound of
this invention means an amount effective to antagonize abnormal
level of CRF or treat the symptoms of affective disorder, anxiety
or depression in a host.
[0049] As used herein, the term "labeled" is meant that the
compound is either directly or indirectly labeled with a label
which provides a detectable signal, e.g. radioisotope, fluorescers,
enzyme, antibodies, particles such as magnetic particles,
chemiluminescer, P.sup.32, I.sup.131, and At.sup.211, etc.
Syntheses
[0050] Many organic compounds exist in optically active forms,
i.e., they have the ability to rotate the plane of plane-polarized
light. In describing an optically active compound, the prefixes D
and L or R and S are used to denote the absolute configuration of
the molecule about its chiral center(s) . The prefixes d and l or
(+) and (-) are employed to designate the sign of rotation of
plane-polarized light by the compound, with (-) or l meaning that
the compound is levorotatory. A compound prefixed with (+) or d is
dextrorotatory. For a given chemical structure, these compounds,
called stereoisomers, are identical except that they are mirror
images of one another. A specific stereoisomer may also be referred
to as an enantiomer, and a mixture of such isomers is often called
an enantiomeric mixture. A 50:50 mixture of enantiomers is referred
to as a racemic mixture.
[0051] The present invention includes all stereoisomeric forms of
the compounds of the formula I. Centers of asymmetry that are
present in the compounds of formula I can all independently of one
another have S configuration or R configuration. The prefixes d and
l or (+) and (-) are employed to designate the sign of rotation of
plane-polarized light by the compound, with (-) or l meaning that
the compound is levorotatory. A compound prefixed with (+) or d is
dextrorotatory. The invention includes all possible enantiomers and
diastereomers and mixtures of two or more stereoisomers, for
example mixtures of enantiomers and/or diastereomers, in all
ratios. Thus, enantiomers are a subject of the invention in
enantiomerically pure form, both as levorotatory and as
dextrorotatory antipodes, in the form of racemates and in the form
of mixtures of the two enantiomers in all ratios. In the case of a
cis/trans isomerism the invention includes both the cis form and
the trans form as well as mixtures of these forms in all ratios.
The preparation of individual stereoisomers can be carried out, if
desired, by separation of a mixture by customary methods, for
example by chromatography or crystallization, by the use of
stereochemically uniform starting materials for the synthesis or by
stereoselective synthesis. optionally a derivatization can be
carried out before a separation of stereoisomers. The separation of
a mixture of stereoisomers can be carried out at the stage of the
compounds of the formula I or at the stage of an intermediate
during the synthesis. The present invention also includes all
tautomeric forms of the compounds of formula (I).
[0052] The compound of Formula (I) may be prepared from using the
procedures outlined in Scheme 1. 3
[0053] A compound of Formula (II), where X=F, may be treated with a
metal alkoxide (e.g. sodium methoxide, potassium methoxide;
pre-formed or generated in situ) in an inert solvent to generate an
intermediate of Formula (III). Inert solvents may include, but are
not limited to, alkyl alcohols (1 to 8 carbons, preferably methanol
or ethanol), lower alkanenitriles (1 to 6 carbons, preferably
acetonitrile), water, dialkyl ethers (preferably diethyl ether),
cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane),
N,N-dialkylformamides (preferably dimethylformamide),
N,N-dialkylacetamides (preferably dimethylacetamide), cyclic amides
(preferably N-methylpyrrolidin-2-one), dialkylsulfoxides
(preferably dimethylsulfoxide) or aromatic hydrocarbons (preferably
benzene or toluene). Preferred reaction temperatures range from
0.degree. C. to 100.degree. C.
[0054] Alternatively, a compound of Formula (II), where X=OH, may
be treated with an alkylating agent in the presence of a base in an
inert solvent to generate an intermediate of Formula (III).
Alkylating agents include, but are not limited to, haloalkanes
(e.g. CH.sub.3I), dialkyl sulfates (e.g. Me.sub.2SO.sub.4) or alkyl
trifluoro-sulfonates (e.g. CH.sub.3O.sub.3SCF.sub.3).
[0055] Bases may include, but are not limited to, alkali metals,
alkali metal hydrides (preferably sodium hydride), alkali metal
alkoxides (1 to 6 carbons) (preferably sodium methoxide or sodium
ethoxide), alkaline earth metal hydrides, alkali metal carbonates,
alkaline metal carbonates, transition metal carbonates (e.g. silver
carbonate), alkali metal dialkylamides (preferably lithium
di-isopropylamide), alkali metal bicarbonates, alkali metal
hydroxides, alkali metal bis(trialkylsilyl)amides (preferably
sodium bis(trimethylsilyl)amide), trialkyl amines (preferably
N,N-di-isopropyl-N-ethyl amine) or aromatic amines (preferably
pyridine).
[0056] Inert solvents may include, but are not limited to,
halocarbons (1 to 8 carbons, 1 to 8 halogens), lower alkanenitriles
(1 to 6 carbons, preferably acetonitrile), water, dialkyl ethers
(preferably diethyl ether), cyclic ethers (preferably
tetrahydrofuran or 1,4-dioxane), N,N-dialkylformamides (preferably
dimethylformamide), N,N-dialkylacetamides (preferably
dimethylacetamide), cyclic amides (preferably
N-methylpyrrolidin-2-one), dialkylsulfoxides (preferably
dimethylsulfoxide) or aromatic hydrocarbons (preferably benzene or
toluene). Preferred reaction temperatures range from 50.degree. C.
to 150.degree. C.
[0057] A compound of Formula (III) may be transformed to a compound
of Formula (IV) by reaction with a brominating agent in the
presence or absence of an additive in an inert solvent. Brominating
agents include, but are not limited to,
N-bromosuccinimide-2,2'-azobisisobutyro-nitrile (AIBN),
N-bromophthalimide-2,2'-azobisiso-butyronitrile (AIBN)), bromine.
Additives include, but are not limited to, alkali metal phosphates
(e.g. K.sub.3PO.sub.4, Na.sub.3PO.sub.4), alkali metal hydrogen
phosphates (e.g. Na.sub.2HPO.sub.4, K.sub.2HPO.sub.4), alkali metal
dihydrogen phosphates (e.g. NaH.sub.2PO.sub.4, KH.sub.2PO.sub.4).
Inert solvents include, but are not limited to, halocarbons (1 to 6
carbons, 1 to 6 halogens (preferably chlorine), water,
N,N-dialkylformamides (preferably dimethylformamide),
N,N-dialkylacetamides (preferably dimethylacetamide), cyclic amides
(preferably N-methylpyrrolidin-2-one). Reaction temperatures range
from 0.degree. C. to 200.degree. C. (preferably 20.degree. C. to
120.degree. C).
[0058] A compound of Formula (IV) may be converted to a compound of
Formula (V) by sequential reactions with (1) an alkyl lithium in an
inert solvent at temperatures ranging from -100.degree. C. to
50.degree. C.; (2) a compound of the Formula B(OR.sup.a).sub.3
(where R.sup.a is branched or straight chain alkyl of 1 to 20
carbons) at temperatures ranging from -100.degree. C. to 50.degree.
C. and (3) an acid in the presence or absence of water at
temperatures ranging from -100.degree. C. to 100.degree. C. Alkyl
lithiums may be branched or straight chain compounds containing 1
to 20 carbons. Inert solvents include, but are not limited to,
dialkyl ethers (preferably diethyl ether), cyclic ethers
(preferably tetrahydrofuran or 1,4-dioxane), or aromatic
hydrocarbons (preferably benzene or toluene).
[0059] Acids may include, but are not limited to, alkanoic acids of
2 to 10 carbons (preferably acetic acid), haloalkanoic acids (2-10
carbons, 1-10 halogens, such as trifluoroacetic acid), arylsulfonic
acids (preferably p-toluenesulfonic acid or benzenesulfonic acid),
alkanesulfonic acids of 1 to 10 carbons (preferably methanesulfonic
acid), hydrochloric acid, sulfuric acid or phosphoric acid.
[0060] A compound of Formula (VII) may be produced by reaction of a
compound of Formula (V) with a compound of Formula (VI) in the
presence of a complex or salt of palladium or nickel, a base and an
inert solvent. Complexes of palladium or nickel include, but are
not limited to, phosphine complexes such as Pd(PPh.sub.3).sub.4,
PdCl.sub.2(PPh.sub.3).su- b.2, NiCl.sub.2(PPh.sub.3).sub.2, or
[1,1-bis(diphenylphosphino)ferrocene]- -dichloropalladium. Bases
may include, but are not limited to, alkali metals, alkali metal
hydrides (preferably sodium hydride), alkali metal alkoxides (1 to
6 carbons) (preferably sodium methoxide or sodium ethoxide), alkali
metal carbonates, alkaline metal carbonates (e.g. barium
carbonate), transition metal carbonates (e.g. silver carbonate) or
trialkyl amines (e.g. triethyl amine). Inert solvents may include,
but are not limited to, dialkyl ethers (preferably diethyl ether),
cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane), or
aromatic hydrocarbons (preferably benzene or toluene). Preferred
reaction temperatures range from -100.degree. C. to 100.degree.
C.
[0061] An intermediate of Formula (VII) may be reacted with a base
in the presence of an inert solvent to afford a compound of Formula
(VIII), where M is an alkali metal cation (e.g. sodium or
potassium). Bases may include, but are not limited to, alkali metal
hydroxides (e.g. NaOH or KOH), alkali metal alkoxides (1 to 6
carbons)(preferably sodium methoxide or sodium ethoxide) or
alkaline earth metal hydroxides. Inert solvents may include, but
are not limited to, alkyl alcohols (1 to 6 carbons), lower
alkanenitriles (1 to 6 carbons, preferably acetonitrile), water,
cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane),
N,N-dialkylformamides (preferably dimethylformamide),
N,N-dialkylacetamides (preferably dimethylacetamide), cyclic amides
(preferably N-methylpyrrolidin-2-one), dialkylsulfoxides
(preferably dimethylsulfoxide). Preferred reaction temperatures
range from 0.degree. C. to 150.degree. C.
[0062] Compounds of Formula (VIII) may be treated with
hydrazine-hydrate in the presence of an acid and an inert solvent
at temperatures ranging from 0.degree. C. to 200.degree. C.,
preferably 70.degree. C. to 150.degree. C., to produce compounds of
Formula (IX). Acids may include, but are not limited to, alkanoic
acids of 2 to 10 carbons (preferably acetic acid), haloalkanoic
acids (2-10 carbons, 1-10 halogens, such as trifluoroacetic acid),
arylsulfonic acids (preferably p-toluenesulfonic acid or
benzenesulfonic acid), alkanesulfonic acids of 1 to 10 carbons
(preferably methanesulfonic acid), hydrochloric acid, sulfuric acid
or phosphoric acid.
[0063] Inert solvents may include, but are not limited to, water,
alkyl alcohols (1 to 8 carbons, preferably methanol or ethanol),
lower alkanenitriles (1 to 6 carbons, preferably acetonitrile),
cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane),
N,N-dialkylformamides (preferably dimethylformamide),
N,N-dialkylacetamides (preferably dimethylacetamide), cyclic amides
(preferably N-methylpyrrolidin-2-one), dialkylsulfoxides
(preferably dimethylsulfoxide) or aromatic hydrocarbons (preferably
benzene or toluene).
[0064] A compound of Formula (IX) may be reacted with compounds of
Formula H.sub.3C(C.dbd.NH)OR.sup.C (where R.sup.C is alkyl (1-6
carbons)) in the presence or absence of an acid in the presence of
an inert solvent at temperatures ranging from 0.degree. C. to
200.degree. C. to produce a compound of Formula (X). Acids may
include, but are not limited to alkanoic acids of 2 to 10 carbons
(preferably acetic acid), haloalkanoic acids (2-10 carbons, 1-10
halogens, such as trifluoroacetic acid), arylsulfonic acids
(preferably p-toluenesulfonic acid or benzenesulfonic acid),
alkanesulfonic acids of 1 to 10 carbons (preferably methanesulfonic
acid), hydrochloric acid, sulfuric acid or phosphoric acid.
Stoichiometric or catalytic amounts of such acids may be used.
[0065] Inert solvents may include, but are not limited to, water,
alkanenitriles (1 to 6 carbons, preferably acetonitrile),
halocarbons of 1 to 6 carbons and 1 to 6 halogens (preferably
dichloroethane or chloroform), alkyl alcohols of 1 to 10 carbons
(preferably ethanol), dialkyl ethers (4 to 12 carbons, preferably
diethyl ether or di-isopropylether) or cyclic ethers such as dioxan
or tetrahydrofuran. Preferred temperatures range from 0.degree. C.
to 100.degree. C.
[0066] A compound of Formula (X) may be converted to an
intermediate compound of Formula (XI) by treatment with compounds
C.dbd.O(R.sup.d).sub.2 (where R.sup.d is halogen (preferably
chlorine), alkoxy (1 to 4 carbons) or alkylthio (1 to 4 carbons))
in the presence or absence of a base in an inert solvent at
reaction temperatures from -50.degree. C. to 200.degree. C. Bases
may include, but are not limited to, alkali metal hydrides
(preferably sodium hydride), alkali metal alkoxides (1 to 6
carbons) (preferably sodium methoxide or sodium ethoxide), alkali
metal carbonates, alkali metal hydroxides, trialkyl amines
(preferably N,N-di-isopropyl-N-ethyl amine or triethylamine) or
aromatic amines (preferably pyridine).
[0067] Inert solvents may include, but are not limited to, alkyl
alcohols (1 to 8 carbons, preferably methanol or ethanol), lower
alkanenitriles (1 to 6 carbons, preferably acetonitrile), cyclic
ethers (preferably tetrahydrofuran or 1,4-dioxane),
N,N-dialkylformamides (preferably dimethylformamide),
N,N-dialkylacetamides (preferably dimethylacetamide), cyclic amides
(preferably N-methylpyrrolidin-2-one), dialkylsulfoxides
(preferably dimethylsulfoxide) or aromatic hydrocarbons (preferably
benzene or toluene).
[0068] A compound of Formula (XI) may be treated with a
halogenating agent in the presence or absence of a base in the
presence or absence of an inert solvent at reaction temperatures
ranging from -80.degree. C. to 250.degree. C. to give a halogenated
intermediate (XII) (where X is halogen). Halogenating agents
include, but are not limited to, SOCl.sub.2, POCl.sub.3, PCl.sub.3,
PCl.sub.5, POBr.sub.3, PBr.sub.3 or PBr.sub.5. Bases may include,
but are not limited to, trialkyl amines (preferably
N,N-di-isopropyl-N-ethyl amine or triethylamine) or aromatic amines
(preferably N,N-diethylaniline).
[0069] Inert solvents may include, but are not limited to,
N,N-dialkylformamides (preferably dimethylformamide),
N,N-dialkylacetamides (preferably dimethylacetamide), cyclic amides
(preferably N-methylpyrrolidin-2-one) or aromatic hydrocarbons
(preferably benzene or toluene) Preferred reaction temperatures
range from 20.degree. C. to 200.degree. C.
[0070] A compound of Formula (XII) may be reacted with an alkyl
amine in the presence or absence of a base in the presence or
absence of an inert solvent at reaction temperatures ranging from
-80 to 250.degree. C. to generate compounds of Formula (I). Bases
may include, but are not limited to, alkali metal hydrides
(preferably sodium hydride), alkali metal alkoxides (1 to 6
carbons) (preferably sodium methoxide or sodium ethoxide), alkaline
earth metal hydrides, alkali metal dialkylamides (preferably
lithium di-isopropylamide), alkali metal carbonates, alkali metal
bicarbonates, alkali metal bis(trialkylsilyl)amides (preferably
sodium bis(trimethylsilyl)amide), trialkyl amines (preferably
N,N-di-isopropyl-N-ethyl amine) or aromatic amines (preferably
pyridine).
[0071] Inert solvents may include, but are not limited to, alkyl
alcohols (1 to 8 carbons, preferably methanol or ethanol), lower
alkanenitriles (1 to 6 carbons, preferably acetonitrile), dialkyl
ethers (preferably diethyl ether), cyclic ethers (preferably
tetrahydrofuran or 1,4-dioxane), N,N-dialkylformamides (preferably
dimethylformamide), N,N-dialkylacetamides (preferably
dimethylacetamide), cyclic amides (preferably
N-methylpyrrolidin-2-one), dialkylsulfoxides (preferably
dimethylsulfoxide), aromatic hydrocarbons (preferably benzene or
toluene) or haloalkanes of 1 to 10 carbons and 1 to 10 halogens
(preferably dichloroethane). Preferred reaction temperatures range
from 0.degree. C. to 140.degree. C.
[0072] The compounds of the invention may be prepared as
radiolabeled compounds 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, Kans.; American
Radiolabeled Chemicals, Inc., St. Louis, Mo.; and Moravek
Biochemicals Inc., Brea, Calif.
[0073] Tritium labeled probe compounds may also conveniently be
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.
[0074] Receptor autoradiography (receptor mapping) may be 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.
EXAMPLES
[0075] Analytical data were recorded for the compounds described
below using the following general procedures. Proton NMR spectra
were recorded on a Varian VXR or Unity 300 FT-NMR instruments (300
MHz); chemical shifts were recorded in ppm (.delta.) from an
internal tetramethysilane standard in deuterochloroform or
deuterodimethylsulfoxide as specified below. Mass spectra (MS) or
high resolution mass spectra (HRMS) were recorded on a Finnegan MAT
8230 spectrometer or a Hewlett Packard 5988A model spectrometer
(using chemi-ionization (CI) with NH.sub.3 as the carrier gas,
electrospray (ESI), atmospheric pressure chemi-ionization (APCI) or
gas chromatography (GC)). Melting points were recorded on a MelTemp
3.0 heating block apparatus and are uncorrected. Boiling points are
uncorrected. All pH determinations during workup were made with
indicator paper.
[0076] Reagents were purchased from commercial sources and, where
necessary, purified prior to use according to the general
procedures outlined by D. Perrin and W. L. F. Armarego,
Purification of Laboratory Chemicals, 3rd ed., (New York: Pergamon
Press, 1988). Chromatography was performed on silica gel using the
solvent systems indicated below. For mixed solvent systems, the
volume ratios are given. Otherwise, parts and percentages are by
weight. Commonly used abbreviations are: DMF
(N,N-dimethylformamide), EtOH (ethanol), MeOH (methanol), EtOAc
(ethyl acetate), HOAc (acetic acid), DME (1,2-diethoxyethane) and
THF (tetrahydrofuran).
[0077] The following examples are provided to describe the
invention in further detail. These examples, which set forth the
best mode presently contemplated for carrying out the invention,
are intended to illustrate and not to limit the invention.
EXAMPLE 1
Preparation of 2,7-dimethyl-8-(2-methyl-6-methoxypyrid-3-yl)
[1,5-a]-pyrazolo-[1,3,5]-triazin-4(3H)-one
[0078] A. 2-Methoxy-6-methylpyridine
[0079] Sodium (31.0 g, 1.35 mol) was added portionwise to methanol
(500 mL) over 30 min with stirring in a flask equipped with a
reflux cindenser. After the addition was complete, the reaction
mixture was allowed to cool to ambient temperature.
2-Fluoro-6-methylpyridine (50 g, 450 mmol) was added portionwise
with stirring. The reaction mixture was then heated to reflux
temperature and stirred for 48 h. The mix was then cooled to
ambient temperature and solvent was removed in vacuo to provide a
yellow oil. The residue was taken up in water (500 mL) and three
extractions with ether (200 mL) were performed. The combined
organic layers were dried over MgSO.sub.4, filtered and solvent was
removed in vacuo from the filtrate to give a yellow liquid:
.sup.1H-NMR(CDCl.sub.3, 300 MHz): .delta. 7.44 (dd, 1H , J=8,7),
6.71 (d, 1H, J=7), 6.53 (d, 1H, J=8), 3.91 (s, 3H), 2.45 (s,
3H).
[0080] B. 2-Methoxy-6-methylpyridine
[0081] A mixture of 2-hydroxy-6-methylpyridine (6.85 g, 62.8 mmol),
silver carbonate (22.5 g, 81.6 mmol), iodomethane (39.1 mL, 628
mmol) and chloroform (200 mL) was stirred at ambient temperature
for 40 h in the dark. The reaction mixture was filtered through
celite. The collected solid was washed with ether. The combined
filtrates were concentrated in vacuo to give a liquid (6.25 g),
which was identical to the product from Part A.
[0082] C. 6-Methoxy-3-bromo-2-methylpyridine
[0083] A mixture of 2-methoxy-6-methylpyridine (17.0 g, 138 mmol)
and a solution of disodium hydrogen phosphate (0.15M in water, 250
mL) was stirred at room temperature. Bromine (7.1 mL, 138 mmol) was
added dropwise over 15 min via an addition funnel. The reaction
mixture was then stirred at room temperature for 4 h. The clear
colorless solution was diluted with water (500 mL) and extracted
with dichloromethane (200 mL) three times. The combined organic
layers were dried over MgSO.sub.4, filtered and solvent was removed
in vacuo from the filtrate to give a yellow liquid. Flash
chromatography on silica gel (EtOAc:hexane::1:20) and removal of
solvent from the desired combined fractions afforded a clear
colorless liquid (15.4 g): .sup.1H-NMR(CDCl.sub.3, 300 MHz):
.delta. 7.60 (d, 1H, J=8), 6.46 (d, 1H, J=8), 3.89 (s, 3H), 2.54
(s, 3H).
[0084] D. 6-Methoxy-2-methylpyridine-3-boronic acid
[0085] A solution of 6-methoxy-3-bromo-2-methylpyridine (59.8 g,
296 mmol) in dry THF (429 mL) was cooled with stirring to
.about.-78.degree. C. under a nitrogen atmosphere. A solution of
n-butyl lithium (2.5 M, 130.4 mL, 326 mmol) in hexane was added
dropwise over 30 min. The reaction mixture was stirred for 3 h at
.about.-78.degree. C. A solution of tri-isopropyl borate (102.7 mL,
445 mmol) in dry THF (100 mL) was added dropwise over 30 min. The
reaction mixture was warmed to ambient temperature with stirring
over 16 h. Acetic acid (37.35 g, 622 mmol), then water (110 mL)
were added to the reaction mixture with stirring. After 2 h, the
layers were separated and the organic layer was concentrated in
vacuo. The residue was taken up in 2-propanol (750 mL) and solvent
was removed on a rotary evaporator (bath temperature
.about.50.degree. C.). The residue was triturated with ether. The
product was collected by filtration and dried in vacuo (48.4 g):
mp>200.degree. C.; .sup.1H-NMR(CD.sub.3OH, 300 MHz): .delta.
7.83 (d, 1H, J=8), 6.56 (d, 1H, J=8), 3.85 (s, 3H), 2.44 (s, 3H);
GC-MS: 168 (M.sup.++H).
[0086] E. 2-Methyl-3-(5-methylisoxazol-4-yl)-6-methoxypyridine
[0087] A mixture of 4-iodo-5-methylisoxazole (18.2 g, 87 mmol),
6-methoxy-2-methylpyridine-3-boronic acid (14.6 g, 87 mmol), sodium
bicarbonate (22.0 g, 262 mmol), water (150 mL) and DME (150 mL) was
degassed three times with stirring by the application of a vacuum
and then introduction of a nitrogen atmosphere.
[1,1-Bis(diphenylphosphino)fe- rrocene]-dichloropalladium (II)
(2.14 g, 2.6 mmol) was added in one portion. The reaction mixture
was degassed as before. The reaction mixture was then stirred at
80.degree. C. for 4 h, then it was cooled to ambient temperature.
Three extractions with EtOAc, drying the combined organic layers
over MgSO.sub.4, filtration and removal of solvent in vacuo
afforded an oil. Flash chromatography (EtOAc:hexane::1:9) and
removal of solvent in vacuo from the desired fractions gave the
product (7.15 g): .sup.1H-NMR(CDCl.sub.3, 300 MHz): .delta. 8.16
(s, 1H), 7.33 (d, 1H, J=8), 6.63 (d, 1H, J=8), 3.95 (s, 3H), 2.35
(s, 6H); APCI.sup.+-MS: 205 (M.sup.++H).
[0088] F. 1-Cyano-l-(2-methyl-6-methoxypyrid-3-yl)propan-2-one,
sodium salt
[0089] A mixture of sodium methoxide (25% w/w, 13 mL, 70 mmol),
2-methyl-3-(5-methylisoxazol-4-yl)-6-methoxypyridine (7.15 g, 35
mmol) and methanol (50 mL) was stirred at room temperature for 16
h. Solvent was removed in vacuo to give a yellow oil. Trituration
with ether, filtration and drying in vacuo afforded the crude
product as a white solid (9.3 g).
[0090] G.
5-Amino-4-(2-methyl-6-methoxypyrid-3-yl)-3-methylpyrazole
[0091] A mixture of
1-cyano-1-(2-methyl-6-methoxypyrid-3-yl)propan-2-one, sodium salt
(9.3 g), hydrazine-hydrate (6 mL, 123.3 mmol) and glacial acetic
acid (150 mL) was stirred at room temperature for 4 h. The reaction
mixture was concentrated in vacuo. The residue was dissolved in 1N
HCl and the resulting solution was extracted with EtOAc two times.
A 1N NaOH solution was added to the aqueous layer until pH=12. The
resulting semi-solution was extracted three times with ethyl
acetate. The combined organic layers were dried over MgSO.sub.4 and
filtered. Solvent was removed in vacuo to give a viscous oil (5.8
g): .sup.1H-NMR (CDCl.sub.3, 300 MHz): 7.37 (d, 2H, J=8), 6.62 (d,
2H, J=8), 3.95 (s, 3H), 2.36 (s, 3H), 2.08 (s, 3H); APCI.sup.+-MS:
219 (M.sup.++H); 260 (M.sup.++CH.sub.3CN).
[0092] H.
5-Acetamidino-4-(2-methyl-6-methoxypyrid-3-yl)-3-methylpyrazole,
acetic acid salt
[0093] Ethyl acetamidate hydrochloride (6.46g, 52.2 mmol) was added
quickly to a rapidly stirred mixture of potassium carbonate (6.95g,
50.0 mol), dichloromethane (60 mL) and water (150 mL). The layers
were separated and the aqueous layer was extracted with
dichloromethane (2.times.60 mL). The combined organic layers were
dried over MgSO.sub.4 and filtered. Solvent was removed by simple
distillation and the pot residue, a clear pale yellow liquid, was
used without further purification.
[0094] Glacial acetic acid (1.0 mL, 17.4 mmol) was added to a
stirred mixture of
5-amino-4-(2-methyl-6-methoxypyrid-3-yl)-3-methylpyrazole (3.8g,
17.4 mmol), ethyl acetamidate free base and dichloromethane (100
mL). The resulting reaction mixture was stirred at room temperature
for 16 h; at the end of which time, it was concentrated in vacuo.
The residue was triturated with ether, the product was filtered and
washed with copious amounts of ether. The white solid was dried in
vacuo (5.4 g): .sup.1H-NMR (CD.sub.3OH, 300 MHz): 7.43 (d, 2H,
J=8), 6.69 (d, 2H, J=8), 4.9 (br s, 2H), 3.93 (s, 3H), 2.31 (s,
3H), 2.24 (s, 3H), 2.13 (s, 3H), 1.88 (s, 3H); APCI.sup.+-MS: 260
(M.sup.++H).
[0095] I.
2,7-dimethyl-8-(2-methyl-6-methoxypyrid-3-yl)[1,5-a]-pyrazolo-[1-
,3,5]-triazin-4(3H)-one
[0096] Sodium pellets (3.9 g, 169 mmol) were added portionwise to
ethanol (200 mL) with vigorous stirring. After all the sodium
reacted,
5-acetamidino-4-(2-methyl-6-methoxypyrid-3-yl)-3-methylpyrazole,
acetic acid salt (5.4 g, 16.9 mmol) and diethyl carbonate ( 16.4
mL, 135.3 mmol) were added. The resulting reaction mixture was
heated to reflux temperature and stirred for 18 hours. The mix was
cooled to room temperature and solvent was removed in vacuo. The
residue was dissolved in water and a 1N HCl solution was added
slowly until pH.about.6. The aqueous layer was extracted with EtOAc
three times; the combined organic layers were dried over MgSO.sub.4
and filtered. Solvent was removed in vacuo to give a solid.
Trituration with ether, filtration and drying in vacuo afforded a
white solid (3.9 g): .sup.1H-NMR (CD.sub.3OH, 300 MHz): 7.49 (d,
2H, J=8), 6.69 (d, 2H, J=8), 3.93 (s, 3H), 2.35 (s, 3H), 2.28 (s,
3H), 2.24 (s, 3H); APCI.sup.+-MS: 286 (M.sup.++H).
EXAMPLE 2
Preparation of
4-((R)-2-butylamino)2,7-dimethyl-8-(2-methyl-6-methoxypyrid- -3-yl)
[1,5-a]-pyrazolo-1,3,5-triazine
[0097] A. 4-Chloro-2,7-dimethyl-8-(2-methyl-6-methoxypyrid-3-yl)
[1,5-a]-pyrazolotriazine
[0098] A mixture of 2,7-dimethyl-8-(2-methyl-6-methoxypyrid-3-yl)
[1,5-a]-pyrazolo-1,3,5-triazin-4-one (Example 1, 3.9 g, 13.7 mmol),
di-isopropyl-ethylamine (9.5 mL, 54.7 mmol), phosphorus oxychloride
(5.1 mL, 54.7 mmol) and toluene (75 mL) was stirred at reflux
temperature for 4 h. The volatiles were removed in vacuo. The
residue was loaded on a pad of silica gel on celite and eluted with
a 1:1 mixture of EtOAc and hexane. Solvent was removed in vacuo
from the filtrate to give an oil.
[0099] B.
4-((R)-2-butylamino)2,7-dimethyl-8-(2-methyl-6-methoxypyrid-3-yl- )
[1,5-a]-pyrazolo-1,3,5-triazine
[0100] A mixture of
4-chloro-2,7-dimethyl-8-(2-methyl-6-methoxypyrid-3-yl)
[1,5-a]-pyrazolotriazine, (R)-2-butylamine (2.0 mL, 20.5 mmol),
di-isopropyl-ethylamine (9.5 mL, 54.7 mmol) and dry THF (25 mL) was
stirred at ambient temperature for 18 hours. Solvent was removed in
vacuo. Column chromatography of the residue (first using
EtOAc:hexane::1:2, then using EtOAc:hexane::1:4) afforded the
product. Removal of solvent in vacuo gave a white solid (2.3 g):
mp=118.3.degree. C. ; .sup.1H-NMR (CDCl.sub.3, 300 MHz): .delta.
7.41 (d, 1H, J=8), 6.63 (d, 1H, J=8), 6.25 (br d, 1H, J=9),
4.35-4.30 (m, 1H), 3.95 (s, 3H), 2.49 (s, 3H), 2.35 (s, 3H), 2.30
(s, 3H), 1.76-1.66 (m, 2H), 1.34 (d, 3H, J=7), 1.02 (t, 3H, J =7);
.sup.13C-NMR (CDCl.sub.3, 100.52 MHz): .delta. 163.8, 163.0, 155.7,
153.7, 147.8, 146.6, 141.6, 118.5, 107.4, 106.6, 53.3, 48.2, 29.7,
26.1, 22.9, 20.4, 13.1, 10.3; IR (neat, KBr, cm.sup.-1): 3380 (m) ,
3371 (m) , 2968 (m) , 2928 (m) , 2872 (w), 1621 (s), 1588 (s), 1544
(s), 1489 (s), 1460 (s), 1425 (s), 1413 (s) , 1364 (s) , 1346 (m) ,
1304 (s) , 1275 (s), 1247 (s) , 1198 (m) , 1152 (m) , 1134 (m) ,
1112 (m) , 1034 (s), 1003 (m); ESI(+)-HRMS: Calcd for
C.sub.18H.sub.24N.sub.6O: 341.2089; Found: 341.2093 (M.sup.++H).
Anal. Calcd for C.sub.18H.sub.24N.sub.6O: C, 63.51, H, 7.12, N,
24.69; Found: C, 63.67, H, 7.00, N, 24.49 .
Utility
[0101] Rat CRF Receptor Binding Assay for the Evaluation of
Biological Activity
[0102] Receptor binding affinity to rat cortical receptors was
assayed according to the published methods (E. B. De Souza, J.
Neuroscience, 7: 88 (1987).
[0103] Curves of the inhibition of [.sup.125I-Tyr.sup.0]-o-CRF
binding to cell membranes at various dilutions of test drug were
analyzed by the iterative curve fitting program LIGAND [P. J.
Munson and D. Rodbard, Anal. Biochem. 107:220 (1980), which
provides Ki values for inhibition which are then used to assess
biological activity.
[0104] Inhibition of CRF-Stimulated Adenylate Cyclase Activity
[0105] Inhibition of CRF-stimulated adenylate cyclase activity can
be performed as described by G. Battaglia et al. Synapse 1:572
(1987). Briefly, assays are carried out at 37.degree. C. for 10 min
in 200 ml of buffer containing 100 mM Tris-HCl (pH 7.4 at
37.degree. C.), 10 mM MgCl.sub.2, 0.4 mM EGTA, 0.1% BSA, 1 mm
isobutylmethylxanthine (IBMX), 250 units/ml phosphocreatine kinase,
5 mM creatine phosphate, 100 mM guanosine 5'-triphosphate, 100 nM
oCRF, antagonist peptides (concentration range 10.sup.-9 to
10.sup.-6m) and 0.8 mg original wet weight tissue (approximately
40-60 mg protein). Reactions are initiated by the addition of 1 mM
ATP/.sup.32P]ATP (approximately 2-4 mCi/tube) and terminated by the
addition of 100 ml of 50 mM Tris-HCL, 45 mM ATP and 2% sodium
dodecyl sulfate. In order to monitor the recovery of cAMP, 1 .mu.l
of [.sup.3H]cAMP (approximately 40,000 dpm) is added to each tube
prior to separation. The separation of [.sup.32P]cAMP from
[.sup.32P]ATP is performed by sequential elution over Dowex and
alumina columns.
[0106] In Vivo Biological Assay
[0107] The in vivo activity of a compound of the present invention
can be assessed using any one of the biological assays available
and accepted within the art. Illustrative of these tests include
the Acoustic Startle Assay, the Stair Climbing Test, and the
Chronic Administration Assay. These and other models useful for the
testing of compounds of the present invention have been outlined in
C. W. Berridge and A. J. Dunn Brain Research Reviews 15:71
(1990).
[0108] A compound may be tested in any species of rodent or small
mammal.
[0109] A compound of this invention has utility in the treatment of
imbalances associated with abnormal levels of corticotropin
releasing factor in patients suffering from depression, affective
disorders, and/or anxiety.
[0110] A compound of this invention can be administered to treat
these abnormalities by means that produce contact of the active
agent with the agent's site of action in the body of a mammal. The
compounds can be administered by any conventional means available
for use in conjunction with pharmaceuticals either as individual
therapeutic agent or in combination of therapeutic agents. It can
be administered alone, but will generally be administered with a
pharmaceutical carrier selected on the basis of the chosen route of
administration and standard pharmaceutical practice.
[0111] The dosage administered will vary depending on the use and
known factors such as pharmacodynamic character of the particular
agent, and its mode and route of administration; the recipient's
age, weight, and health; nature and extent of symptoms; kind of
concurrent treatment; frequency of treatment; and desired effect.
For use in the treatment of said diseases or conditions, a compound
of this invention can be orally administered daily at a dosage of
the active ingredient of 0.002 to 200 mg/kg of body weight.
Ordinarily, a dose of 0.01 to 10 mg/kg in divided doses one to four
times a day, or in sustained release formulation will be effective
in obtaining the desired pharmacological effect.
[0112] Dosage forms (compositions) suitable for administration
contain from about 1 mg to about 100 mg of active ingredient per
unit. In these pharmaceutical compositions, the active ingredient
will ordinarily be present in an amount of about 0.5 to 95% by
weight based on the total weight of the composition.
[0113] The active ingredient can be administered orally is solid
dosage forms, such as capsules, tablets and powders; or in liquid
forms such as elixirs, syrups, and/or suspensions. The compounds of
this invention can also be administered parenterally in sterile
liquid dose formulations.
[0114] Gelatin capsules can be used to contain the active
ingredient and a suitable carrier such as but not limited to
lactose, starch, magnesium stearate, steric acid, or cellulose
derivatives. Similar diluents can be used to make compressed
tablets. Both tablets and capsules can be manufactured as sustained
release products to provide for continuous release of medication
over a period of time. Compressed tablets can be sugar-coated or
film-coated to mask any unpleasant taste, or used to protect the
active ingredients from the atmosphere, or to allow selective
disintegration of the tablet in the gastrointestinal tract.
[0115] Liquid dose forms for oral administration can contain
coloring or flavoring agents to increase patient acceptance.
[0116] In general, water, pharmaceutically acceptable oils, saline,
aqueous dextrose (glucose), and related sugar solutions and
glycols, such as propylene glycol or polyethylene glycol, are
suitable carriers for parenteral solutions. Solutions for
parenteral administration preferably contain a water soluble salt
of the active ingredient, suitable stabilizing agents, and if
necessary, butter substances. Antioxidizing agents, such as sodium
bisulfite, sodium sulfite, or ascorbic acid, either alone or in
combination, are suitable stabilizing agents. Also used are citric
acid and its salts, and EDTA. In addition, parenteral solutions can
contain preservatives such as benzalkonium chloride, methyl- or
propyl-paraben, and chlorobutanol.
[0117] Suitable pharmaceutical carriers are described in
"Remington's Pharmaceutical Sciences", A. Osol, a standard
reference in the field.
[0118] Useful pharmaceutical dosage-forms for administration of the
compounds of this invention can be illustrated as follows:
[0119] Capsules
[0120] A large number of units capsules are prepared by filling
standard two-piece hard gelatin capsules each with 100 mg of
powdered active ingredient, 150 mg lactose, 50 mg cellulose, and 6
mg magnesium stearate.
[0121] Soft Gelatin Capsules
[0122] A mixture of active ingredient in a digestible oil such as
soybean, cottonseed oil, or olive oil is prepared and injected by
means of a positive displacement was pumped into gelatin to form
soft gelatin capsules containing 100 mg of the active ingredient.
The capsules were washed and dried.
[0123] Tablets
[0124] A large number of tablets are prepared by conventional
procedures so that the dosage unit was 100 mg active ingredient,
0.2 mg of colloidal silicon dioxide, 5 mg of magnesium stearate,
275 mg of microcrystalline cellulose, 11 mg of starch, and 98.8 mg
lactose. Appropriate coatings may be applied to increase
palatability or delayed adsorption.
[0125] The compounds of this invention may also be used as reagents
or standards in the biochemical study of neurological function,
dysfunction, and disease.
[0126] Although the present invention has been described and
exemplified in terms of certain preferred embodiments, other
embodiments will be apparent to those skilled in the art. The
invention is, therefore, not limited to the particular embodiments
described and exemplified, but is capable of modification or
variation without departing from the spirit of the invention, the
full scope of which is delineated by the appended
* * * * *