U.S. patent application number 10/830416 was filed with the patent office on 2004-12-30 for crf receptor antagonists and methods relating thereto.
This patent application is currently assigned to Neurocrine Biosciences, Inc.. Invention is credited to Haddach, Mustapha, Huang, Charles Q., Lanier, Marion C., McCarthy, James R..
Application Number | 20040266799 10/830416 |
Document ID | / |
Family ID | 22928212 |
Filed Date | 2004-12-30 |
United States Patent
Application |
20040266799 |
Kind Code |
A1 |
Haddach, Mustapha ; et
al. |
December 30, 2004 |
CRF receptor antagonists and methods relating thereto
Abstract
CRF receptor antagonists are disclosed which have utility in the
treatment of a variety of disorders, including the treatment of
disorders manifesting hypersecretion of CRF in a warm-blooded
animals, such as stroke.
Inventors: |
Haddach, Mustapha; (San
Diego, CA) ; Lanier, Marion C.; (San Diego, CA)
; Huang, Charles Q.; (San Diego, CA) ; McCarthy,
James R.; (Zionsville, IN) |
Correspondence
Address: |
SEED INTELLECTUAL PROPERTY LAW GROUP PLLC
701 FIFTH AVE
SUITE 6300
SEATTLE
WA
98104-7092
US
|
Assignee: |
Neurocrine Biosciences,
Inc.
San Diego
CA
|
Family ID: |
22928212 |
Appl. No.: |
10/830416 |
Filed: |
April 22, 2004 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
10830416 |
Apr 22, 2004 |
|
|
|
10016694 |
Nov 2, 2001 |
|
|
|
6747034 |
|
|
|
|
60245821 |
Nov 3, 2000 |
|
|
|
Current U.S.
Class: |
514/267 ;
544/250; 544/251 |
Current CPC
Class: |
A61P 25/00 20180101;
A61P 5/04 20180101; A61P 1/00 20180101; A61P 25/22 20180101; A61P
43/00 20180101; C07D 471/06 20130101; A61P 25/24 20180101 |
Class at
Publication: |
514/267 ;
544/250; 544/251 |
International
Class: |
A61K 031/519; C07D
487/14 |
Claims
1. A compound having the following structure: 12including
stereoisomers and pharmaceutically acceptable salts thereof,
wherein: 13represents --N.dbd.CH--, --NH--CH.sub.2-- or
--NH--(CH.sub.2).sub.2--; X is N or CR.sub.3; R.sub.1 is
--CH(R.sub.4)(R.sub.5); R.sub.2 is C.sub.1-6alkyl; R.sub.3 is
hydrogen or C.sub.1-6alkyl; R.sub.4 is hydrogen, C.sub.1-6alkyl,
mono- or di(C.sub.3-6cycloalkyl)methyl, C.sub.3-6cycloalkyl,
C.sub.3-6alkenyl, hydroxyC.sub.1-6alkyl,
C.sub.1-6alkylcarbonyloxyC.sub.1-6alkyl, or
C.sub.1-6alkyloxyC.sub.1-6alk- yl, and R.sub.5 is C.sub.1-8alkyl,
mono- or di(C.sub.3-6cycloalkyl)methyl, Ar.sup.1CH.sub.2,
C.sub.3-6alkenyl, C.sub.1-6alkyloxyC.sub.1-6alkyl,
hydroxyC.sub.1-6alkyl, thienylmethyl, furanylmethyl,
C.sub.1-6alkylthioC.sub.1-6alkyl, morpholinyl, mono- or
di(C.sub.1-6alkyl)aminoC.sub.1-6alkyl, di(C.sub.1-6alkyl)amino,
C.sub.1-6alkylcarbonylC.sub.1-6alkyl, C.sub.1-6alkyl substituted
with imidazolyl, or a radical of the formula
-(C.sub.1-6alkanediyl)-O--CO--Ar.- sup.1, or R.sub.4 and R.sub.5
taken together with the carbon atom to which they are bonded form a
C.sub.5-8cycloalkyl optionally substituted with one or more
substituents independently selected from C.sub.1-6alkyl; Ar is
phenyl substituted with 1, 2 or 3 substituents independently
selected from halo, C.sub.1-6alkyl, trifluoromethyl, cyano,
C.sub.1-6alkyloxy, benzyloxy, C.sub.1-6alkylthio, nitro, amino, and
mono- or di(C.sub.1-6alkyl)amino; or an aromatic
C.sub.3-12heterocycle optionally substituted with 1, 2 or 3
substituents independently selected from halo, C.sub.1-6alkyl,
trifluoromethyl, hydroxy, cyano, C.sub.1-6alkyloxy, benzyloxy,
C.sub.1-6alkylthio, nitro, amino, mono- or di(C.sub.1-6alkyl)amino,
and piperidinyl; and Ar.sup.1 is phenyl, pyridinyl, or phenyl
substituted with 1, 2 or 3 substituents independently selected from
halo, C.sub.1-6alkyl, C.sub.1-6alkyloxy,
di(C.sub.1-6alkyl)aminoC.sub.1-6alkyl, trifluoromethyl and
C.sub.1-6alkyl substituted with morpholinyl.
2. The compound of claim 1 having the structure: 14
3. The compound of claim 1 having the structure: 15
4. The compound of claim 1 having the structure: 16
5. The compound of claim 1 having the structure: 17
6. The compound of claim 1 having the structure: 18
7. The compound of claim 1 having the structure: 19
8. The compound of claim 1 wherein Ar is 2,4-dichlorophenyl.
9. The compound of claim 1 wherein Ar is
2-chloro-4-methyl-phenyl.
10. The compound of claim 1 wherein Ar is
2-methyl-4-chloro-phenyl.
11. The compound of claim 1 wherein Ar is
2,4,6-trimethyl-phenyl.
12. The compound of claim 1 wherein Ar is
2-chloro-4-methoxy-phenyl.
13. The compound of claim 1 wherein Ar is
2-methyl-4-methoxy-phenyl.
14. The compound of claim 1 wherein Ar is 2,4-dimethoxy-phenyl.
15. The compound of claim 1 wherein Ar is
4-dimethylamino-2-methyl-3-pyrid- yl.
16. The compound of claim 1 wherein Ar is
4-dimethylamino-6-methyl-3-pyrid- yl.
17. The compound of claim 1 wherein Ar is
4-dimethylamino-3-pyridyl.
18. The compound of claim 1 wherein R.sub.1 is
--CH(n-propyl).sub.2.
19. The compound of claim 1 wherein R.sub.1 is
--CH(n-propyl)(CH.sub.2OCH.- sub.3).
20. The compound of claim 1 wherein R.sub.1 is
--CH(benzyl)(CH.sub.2OCH.su- b.3).
21. The compound of claim 1 wherein R.sub.1 is
--CH(CH.sub.2OR).sub.2 and each occurrence of R is independently
selected from C.sub.1-6alkyl.
22. The compound of claim 1 wherein R.sub.1 is
--CH(CH.sub.2OR)(ethyl) and each occurrence of R is independently
selected from C.sub.1-6alkyl.
23. The compound of claim 1 wherein R.sub.1 is
--CH(CH.sub.2OR)(n-butyl) and each occurrence of R is independently
selected from C.sub.1-6alkyl.
24. The compound of claim 1 wherein R.sub.1 is
--CH(CH.sub.2OR)(tert-butyl- ) and each occurrence of R is
independently selected from C.sub.1-6alkyl.
25. The compound of claim 1 wherein R.sub.1 is
--CH(CH.sub.2OR)(4-chloro-b- enzyl) and each occurrence of R is
independently selected from C.sub.1-6alkyl.
26. The compound of claim 1 wherein R.sub.1 is
--CH(CH.sub.2OR)(CH.sub.2CH- .sub.2SCH.sub.3) and each occurrence
of R is independently selected from C.sub.1-6alkyl.
27. The compound of claim 1 wherein R.sub.1
--CH(CH.sub.2CH.sub.3)(CH.sub.- 2Obenzyl).
28. The compound of claim 1 wherein R.sub.2 is methyl.
29. The compound of claim 1 wherein R.sub.2 is ethyl.
30. A pharmaceutical composition comprising a compound of claim 1
in combination with a pharmaceutically acceptable carrier or
diluent.
31. A method for treating a disorder manifesting hypersecretion of
CRF in a warm-blooded animal, comprising administering to the
animal an effective amount of the pharmaceutical composition of
claim 30.
32. The method of claim 31 wherein the disorder is stroke.
33. The method of claim 31 wherein the disorder is anxiety.
34. The method of claim 31 wherein the disorder is depression.
35. The method of claim 31 wherein the disorder is irritable bowel
syndrome.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 10/016,694 filed Nov. 2, 2001, which claims the benefit of U.S.
Provisional Application No. 60/245,821 filed Nov. 3, 2000; both of
these applications are incorporated herein by reference in their
entireties.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] This invention relates generally to CRF receptor
antagonists, and to methods of treating disorders by administration
of such antagonists to a warm-blooded animal in need thereof.
[0004] 2. Description of the Related Art
[0005] The first corticotropin-releasing factor (CRF) was isolated
from ovine hypothalmi and identified as a 41-amino acid peptide
(Vale et al., Science 213:1394-1397, 1981). Subsequently, sequences
of human and rat CRF were isolated and determined to be identical,
but different from ovine CRF in 7 of the 41 amino acid residues
(Rivier et al., Proc. Natl. Acad. Sci. USA 80:4851, 1983;
Shibaharaetal., EMBO J 2:775, 1983).
[0006] CRF has been found to produce profound alterations in
endocrine, nervous and immune system function. CRF is believed to
be the major physiological regulator of the basal and
stress-release of adrenocorticotropic hormone ("ACTH"),
.beta.-endorphin, and other pro-opiomelanocortin ("POMC")-derived
peptides from the anterior pituitary (Vale et al., Science
213:1394-1397, 1981). Briefly, CRF is believed to initiate its
biological effects by binding to a plasma membrane receptor which
has been found to be distributed throughout the brain (DeSouza et
al., Science 224:1449-1451, 1984), pituitary (DeSouza et al.,
Methods Enzymol. 124:560, 1986; Wynn et al., Biochem. Biophys. Res.
Comm. 110:602-608, 1983), adrenals (Udelsman et al., Nature
319:147-150, 1986) and spleen (Webster, E. L., and E. B. DeSouza,
Endocrinology 122:609-617, 1988). The CRF receptor is coupled to a
GTP-binding protein (Perrin et al., Endocrinology 118:1171-1179,
1986) which mediates CRF-stimulated increase in intracellular
production of cAMP (Bilezikjian, L. M., and W. W. Vale,
Endocrinology 113:657-662, 1983). The receptor for CRF has now been
cloned from rat (Perrin et al., Endo 133(6):3058-3061, 1993), and
human brain (Chen et al., PNAS 90(19):8967-8971, 1993; Vita et al.,
FEBS 335(1):1-5, 1993). This receptor is a 415 amino acid protein
comprising seven membrane spanning domains. A comparison of
identity between rat and human sequences shows a high degree of
homology (97%) at the amino acid level.
[0007] In addition to its role in stimulating the production of
ACTH and POMC, CRF is also believed to coordinate many of the
endocrine, autonomic, and behavioral responses to stress, and may
be involved in the pathophysiology of affective disorders.
Moreover, CRF is believed to be a key intermediary in communication
between the immune, central nervous, endocrine and cardiovascular
systems (Crofford et al., J. Clin. Invest. 90:2555-2564, 1992;
Sapolsky et al., Science 238:522-524, 1987; Tilders et al., Regul.
Peptides 5:77-84, 1982). Overall, CRF appears to be one of the
pivotal central nervous system neurotransmitters and plays a
crucial role in integrating the body's overall response to
stress.
[0008] Administration of CRF directly to the brain elicits
behavioral, physiological, and endocrine responses identical to
those observed for an animal exposed to a stressful environment.
For example, intracerebroventricular injection of CRF results in
behavioral activation (Sutton et al., Nature 297:331, 1982),
persistent activation of the electroencephalogram (Ehlers et al.,
Brain Res. 278:332, 1983), stimulation of the
sympathoadrenomedullary pathway (Brown et al., Endocrinology
110:928, 1982), an increase of heart rate and blood pressure
(Fisher et al., Endocrinology 110:2222, 1982), an increase in
oxygen consumption (Brown et al., Life Sciences 30:207, 1982),
alteration of gastrointestinal activity (Williams et al., Am. J.
Physiol. 253:G582, 1987), suppression of food consumption (Levine
et al., Neuropharmacology 22:337, 1983), modification of sexual
behavior (Sirinathsinghji et al., Nature 305:232, 1983), and immune
function compromise (Irwin et al., Am. J. Physiol. 255:R744, 1988).
Furthermore, clinical data suggests that CRF may be hypersecreted
in the brain in depression, anxiety-related disorders, and anorexia
nervosa. (DeSouza, Ann. Reports in Med. Chem. 25:215-223, 1990).
Accordingly, clinical data suggests that CRF receptor antagonists
may represent novel antidepressant and/or anxiolytic drugs that may
be useful in the treatment of the neuropsychiatric disorders
manifesting hypersecretion of CRF.
[0009] The first CRF receptor antagonists were peptides (see, e.g.,
Rivier et al., U.S. Pat. No. 4,605,642; Rivier et al., Science
224:889, 1984). While these peptides established that CRF receptor
antagonists can attenuate the pharmacological responses to CRF,
peptide CRF receptor antagonists suffer from the usual drawbacks of
peptide therapeutics including lack of stability and limited oral
activity. More recently, small molecule CRF receptor antagonists
have been reported. For example, substituted
4-thio-5-oxo-3-pyyrazoline derivatives (Abreu et al., U.S. Pat. No.
5,063,245) and substituted 2-aminothiazole derivatives
(Courtemanche et al., Australian Patent No. AU-A-41399/93) have
been reported as CRF receptor antagonists. These particular
derivatives were found to be effective in inhibiting the binding of
CRF to its receptor in the 1-10 .mu.M range and 0.1-10 .mu.M range,
respectively.
[0010] Due to the physiological significance of CRF, the
development of biologically-active small molecules having
significant CRF receptor binding activity and which are capable of
antagonizing the CRF receptor remains a desirable goal. Such CRF
receptor antagonists would be useful in the treatment of endocrine,
psychiatric and neurologic conditions or illnesses, including
stress-related disorders in general.
[0011] While significant strides have been made toward achieving
CRF regulation through administration of CRF receptor antagonists,
there remains a need in the art for effective small molecule CRF
receptor antagonists. There is also a need for pharmaceutical
compositions containing such CRF receptor antagonists, as well as
methods relating to the use thereof to treat, for example,
stress-related disorders. The present invention fulfills these
needs, and provides other related advantages.
SUMMARY OF THE INVENTION
[0012] In brief, this invention is generally directed to CRF
receptor antagonists, and more specifically to CRF receptor
antagonists having the following general structure (I): 1
[0013] including stereoisomers and pharmaceutically acceptable
salts thereof, wherein X, R.sub.1, R.sub.2 and Ar are as defined
below.
[0014] The CRF receptor antagonists of this invention have utility
over a wide range of therapeutic applications, and may be used to
treat a variety of disorders or illnesses, including stress-related
disorders. Such methods include administering an effective amount
of a CRF receptor antagonist of this invention, preferably in the
form of a pharmaceutical composition, to an animal in need thereof.
Accordingly, in another embodiment, pharmaceutical compositions are
disclosed containing one or more CRF receptor antagonists of this
invention in combination with a pharmaceutically acceptable carrier
and/or diluent.
[0015] These and other aspects of the invention will be apparent
upon reference to the following detailed description. To this end,
various references are set forth herein which describe in more
detail certain procedures, compounds and/or compositions, and are
hereby incorporated by reference in their entirety.
DETAILED DESCRIPTION OF THE INVENTION
[0016] The present invention is directed generally to compounds
useful as corticotropin-releasing factor (CRF) receptor
antagonists.
[0017] In a first embodiment, the CRF receptor antagonists of this
invention have the following structure (I): 2
[0018] including stereoisomers and pharmaceutically acceptable
salts thereof,
[0019] wherein: 3
[0020] represents --N.dbd.CH--, --NH--CH.sub.2-- or
--NH--(CH.sub.2).sub.2--;
[0021] X is N or CR.sub.3;
[0022] R.sub.1 is --CH(R.sub.4)(R.sub.5);
[0023] R.sub.2 is C.sub.1-6alkyl;
[0024] R.sub.3 is hydrogen or C.sub.1-6alkyl;
[0025] R.sub.4 is hydrogen, C.sub.1-6alkyl, mono- or
di(C.sub.3-6cycloalkyl)methyl, C.sub.3-6cycloalkyl,
C.sub.3-6alkenyl, hydroxyC.sub.1-6alkyl,
C.sub.1-6alkylcarbonyloxyC.sub.1-6alkyl, or
C.sub.1-6alkyloxyC.sub.1-6alkyl, and
[0026] R.sub.5 is C.sub.1-8alkyl, mono- or
di(C.sub.3-6cycloalkyl)methyl, Ar.sup.1CH.sub.2, C.sub.3-6alkenyl,
C.sub.1-6alkyloxyC.sub.1-6alkyl, hydroxyC.sub.1-6alkyl,
thienylmethyl, furanylmethyl, C.sub.1-6alkylthioC.sub.1-6alkyl,
morpholinyl, mono- or di(C.sub.1-6alkyl)aminoC.sub.1-6alkyl,
di(C.sub.1-6alkyl)amino, C.sub.1-6alkylcarbonylC.sub.1-6alkyl,
C.sub.1-6alkyl substituted with imidazolyl, or a radical of the
formula --(C.sub.1-6alkanediyl)-O--CO--Ar- .sup.1,
[0027] or R.sub.4 and R.sub.5 taken together with the carbon atom
to which they are bonded form a C.sub.5-8cycloalkyl optionally
substituted with one or more substituents independently selected
from C.sub.1-6alkyl;
[0028] Ar is phenyl substituted with 1, 2 or 3 substituents
independently selected from halo, C.sub.1-6alkyl, trifluoromethyl,
cyano, C.sub.1-6alkyloxy, benzyloxy, C.sub.1-6alkylthio, nitro,
amino, and mono- or di(C.sub.1-6alkyl)amino; or an aromatic
C.sub.3-12heterocycle optionally substituted with 1, 2 or 3
substituents independently selected from halo, C.sub.1-6alkyl,
trifluoromethyl, hydroxy, cyano, C.sub.1-6alkyloxy, benzyloxy,
C.sub.1-6alkylthio, nitro, amino, mono- or di(C.sub.1-6alkyl)amino,
and piperidinyl; and
[0029] Ar.sup.1 is phenyl, pyridinyl, or phenyl substituted with 1,
2 or 3 substituents independently selected from halo,
C.sub.1-6alkyl, C.sub.1-6alkyloxy,
di(C.sub.1-6alkyl)aminoC.sub.1-6alkyl, trifluoromethyl and
C.sub.1-6alkyl substituted with morpholinyl.
[0030] In the context of this invention, the preceding terms have
the meanings set forth below.
[0031] "C.sub.1-6alkyl" or "C.sub.1-8alkyl" represents a straight
chain or branched alkyl having from 1 to 6 carbon atoms or 1 to 8
carbon atoms, respectively, such as methyl, ethyl, n-propyl,
isopropyl, n-butyl, tert-butyl, n-pentyl, and the like.
[0032] "C.sub.1-6alkyloxy" represents the group
--O(C.sub.1-6alkyl).
[0033] "C.sub.1-6alkylthio" represents the group
--S(C.sub.1-6alkyl).
[0034] "C.sub.3-6cycloalkyl" represents a cyclic alkyl having from
3 to 6 carbon atoms, including cyclopropyl, cyclopentyl,
cyclopentyl, and cyclohexyl.
[0035] "C.sub.5-8cycloalkyl" represents a cyclic alkyl having from
5 to 8 carbon atoms, such as cyclopentyl, cyclohexyl, and the
like.
[0036] "C.sub.3-6alkenyl" represents an unsaturated straight chain
or branched alkyl having from 3 to 6 carbon atoms, and having at
least one double bond, such as propylene, 1-butene, 2-butene,
2-methylpropene, and the like.
[0037] "HydroxyC.sub.1-6alkyl" represents a C.sub.1-6alkyl
substituted with at least one hydroxyl group.
[0038] "Mono- or di(C.sub.3-6cycloalkyl)methyl" represents a methyl
group substituted with one or two C.sub.3-6cycloalkyl groups, such
as cyclopropylmethyl, dicyclopropylmethyl, and the like.
[0039] "C.sub.1-6alkylcarbonylC.sub.1-6alkyl" represents a
C.sub.1-6alkyl substituted with a --COC.sub.1-6alkyl group.
[0040] "C.sub.1-6alkylcarbonyloxyC.sub.1-6alkyl" represents a
C.sub.1-6alkyl substituted with a --COOC.sub.1-6alkyl group.
[0041] "C.sub.1-6alkyloxyC.sub.1-6alkyl" represents a
C.sub.1-6alkyl substituted with a --OC.sub.1-6alkyl group.
[0042] "C.sub.1-6alkylthioC.sub.1-6alkyl" represents a
C.sub.1-6alkyl substituted with a --SC.sub.1-6alkyl group.
[0043] "Mono- or di(C.sub.1-6alkyl)amino represents an amino
substituted with one C.sub.1-6alkyl or with two C.sub.1-6alkyls,
respectively.
[0044] "Mono- or di(C.sub.1-6alkyl)aminoC.sub.1-6alkyl" represents
a C.sub.1-6alkyl substituted with a mono- or
di(C.sub.1-6alkyl)amino.
[0045] "C.sub.1-6alkanediyl" represents a divalent C.sub.1-6alkyl
radical, such as methylene (--CH.sub.2--), ethylene
(--CH.sub.2CH.sub.2--), and the like.
[0046] "C.sub.3-12heterocycle" represents a ring made up of more
than one kind of atom, and which contains 3 to 12 carbon atoms,
such as pyridinyl, pyrimidinyl, furanyl, thienyl, imidazolyl,
thiazolyl, pyrazolyl, pyridazinyl, pyrazinyl, triazinyl (such as
1,3,5), and the like.
[0047] "Halo" means fluoro, chloro, bromo or iodo.
[0048] Representative CRF receptor antagonists of this invention
include compounds having the following structures (Ia), (Ib) and
(Ic): 4
[0049] When X of compounds (Ia), (Ib) and (Ic) is N, representative
compounds of this invention include the following compounds (Ia'),
(Ib') and (Ic'), and when X is CR.sub.3, and R.sub.3 is hydrogen,
representative compounds of this invention include the following
compounds (Ia"), (Ib") and (Ic"): 5
[0050] In the embodiment where the R.sub.4 and R.sub.5 groups of
R.sub.1 taken together form a C.sub.5-8cycloalkyl, the resulting
R.sub.1 group has the structure: 6
[0051] When the above structure is optionally substituted with one
or more C.sub.1-6alkyl groups, a representative R.sub.1 moiety has
the following structure: 7
[0052] wherein R.sub.6 and R.sub.7 are the same or different and
independently selected from a C.sub.1-6alkyl, such as methyl or
ethyl.
[0053] Representative Ar, R.sub.1 and R.sub.2 groups of this
invention are set forth in the following Table. To this end, it
should be understood that each combination of the Ar, R.sub.1 and
R.sub.2 groups listed in the following Table 1 represents
individual compounds of structure (I), as well as the more specific
structures (Ia), (Ib) and (Ic) and sub-structures (Ia'), (Ia"),
(Ib'), (Ib"), (Ic') and (Ic").
1TABLE 1 REPRESENTATIVE AR, R.sub.1 AND R.sub.2 GROUPS OF STRUCTURE
(I) Ar R.sub.1 R.sub.2 2,4-dichlorophenyl --CH(n-propyl).sub.2
--CH.sub.3 2-chloro-4-methyl-phenyl --CH(n-propyl).sub.2 --CH.sub.3
2-methyl-4-chloro-phenyl --CH(n-propyl).sub.2 --CH.sub.3
2,4,6-trimethyl-phenyl --CH(n-propyl).sub.2 --CH.sub.3
2-chloro-4-methoxy-phenyl --CH(n-propyl).sub.2 --CH.sub.3
2-methyl-4-methoxy-phenyl --CH(n-propyl).sub.2 --CH.sub.3
2,4-dimethoxy-phenyl --CH(n-propyl).sub.2 --CH.sub.3
4-dimethylamino-2-methyl- --CH(n-propyl).sub.2 --CH.sub.3 3-pyridyl
4-dimethylamino-6-methyl- --CH(n-propyl).sub.2 --CH.sub.3 3-pyridyl
4-dimethylamino-3-pyridyl --CH(n-propyl).sub.2 --CH.sub.3
2,4-dichlorophenyl --CH(n-propyl)(CH.sub.2OCH.sub.3) --CH.sub.3
2-chloro-4-methyl-phenyl --CH(n-propyl)(CH.sub.2OCH.sub.3)
--CH.sub.3 2-methyl-4-chloro-phenyl
--CH(n-propyl)(CH.sub.2OCH.sub.3) --CH.sub.3 2,4,6-trimethyl-phenyl
--CH(n-propyl)(CH.sub.2OCH.sub.3) --CH.sub.3
2-chloro-4-methoxy-phenyl --CH(n-propyl)(CH.sub.2OCH.su- b.3)
--CH.sub.3 2-methyl-4-methoxy-phenyl --CH(n-propyl)(CH.sub.2OC-
H.sub.3) --CH.sub.3 2,4-dimethoxy-phenyl
--CH(n-propyl)(CH.sub.2OCH- .sub.3) --CH.sub.3
4-dimethylamino-2-methyl- --CH(n-propyl)(CH.sub.2OCH.sub.3)
--CH.sub.3 3-pyridyl 4-dimethylamino-6-methyl-
--CH(n-propyl)(CH.sub.2OCH.sub.3) --CH.sub.3 3-pyridyl
4-dimethylamino-3-pyridyl --CH(n-propyl)(CH.sub.2OCH- .sub.3)
--CH.sub.3 2,4-dichlorophenyl --CH(benzyl)(CH.sub.2OCH.sub.- 3)
--CH.sub.3 2-chloro-4-methyl-phenyl --CH(benzyl)(CH.sub.2OCH.sub-
.3) --CH.sub.3 2-methyl-4-chloro-phenyl
--CH(benzyl)(CH.sub.2OCH.su- b.3) --CH.sub.3 2,4,6-trimethyl-phenyl
--CH(benzyl)(CH.sub.2OCH.sub- .3) --CH.sub.3
2-chloro-4-methoxy-phenyl --CH(benzyl)(CH.sub.2OCH.s- ub.3)
--CH.sub.3 2-methyl-4-methoxy-phenyl --CH(benzyl)(CH.sub.2OCH-
.sub.3) --CH.sub.3 2,4-dimethoxy-phenyl
--CH(benzyl)(CH.sub.2OCH.su- b.3) --CH.sub.3
4-dimethylamino-2-methyl- --CH(benzyl)(CH.sub.2OCH.- sub.3)
--CH.sub.3 3-pyridyl 4-dimethylamino-6-methyl-
--CH(benzyl)(CH.sub.2OCH.sub.3) --CH.sub.3 3-pyridyl
4-dimethylamino-3-pyridyl --CH(benzyl)(CH.sub.2OCH.sub.3)
--CH.sub.3 2,4-dichlorophenyl --CH(CH.sub.2OR).sub.2 --CH.sub.3
2-chloro-4-methyl-phenyl --CH(CH.sub.2OR).sub.2 --CH.sub.3
2-methyl-4-chloro-phenyl --CH(CH.sub.2OR).sub.2 --CH.sub.3
2,4,6-trimethyl-phenyl --CH(CH.sub.2OR).sub.2 --CH.sub.3
2-chloro-4-methoxy-phenyl --CH(CH.sub.2OR).sub.2 --CH.sub.3
2-methyl-4-methoxy-phenyl --CH(CH.sub.2OR).sub.2 --CH.sub.3
2,4-dimethoxy-phenyl --CH(CH.sub.2OR).sub.2 --CH.sub.3
4-dimethylamino-2-methyl- --CH(CH.sub.2OR).sub.2 --CH.sub.3
3-pyridyl 4-dimethylamino-6-methyl- --CH(CH.sub.2OR).sub.2
--CH.sub.3 3-pyridyl 4-dimethylamino-3-pyridyl
--CH(CH.sub.2OR).sub.2 --CH.sub.3 2,4-dichlorophenyl
--CH(CH.sub.2OR)(ethyl) --CH.sub.3 2-chloro-4-methyl-phenyl
--CH(CH.sub.2OR)(ethyl) --CH.sub.3 2-methyl-4-chloro-phenyl
--CH(CH.sub.2OR)(ethyl) --CH.sub.3 2,4,6-trimethyl-phenyl
--CH(CH.sub.2OR)(ethyl) --CH.sub.3 2-chloro-4-methoxy-phenyl
--CH(CH.sub.2OR)(ethyl) --CH.sub.3 2-methyl-4-methoxy-phenyl
--CH(CH.sub.2OR)(ethyl) --CH.sub.3 2,4-dimethoxy-phenyl
--CH(CH.sub.2OR)(ethyl) --CH.sub.3 4-dimethylamino-2-methyl-
--CH(CH.sub.2OR)(ethyl) --CH.sub.3 3-pyridyl
4-dimethylamino-6-methyl- --CH(CH.sub.2OR)(ethyl) --CH.sub.3
3-pyridyl 4-dimethylamino-3-pyridyl --CH(CH.sub.2OR)(ethyl)
--CH.sub.3 2,4-dichlorophenyl --CH(CH.sub.2OR)(n-butyl) --CH.sub.3
2-chloro-4-methyl-phenyl --CH(CH.sub.2OR)(n-butyl) --CH.sub.3
2-methyl-4-chloro-phenyl --CH(CH.sub.2OR)(n-butyl) --CH.sub.3
2,4,6-trimethyl-phenyl --CH(CH.sub.2OR)(n-butyl) --CH.sub.3
2-chloro-4-methoxy-phenyl --CH(CH.sub.2OR)(n-butyl) --CH.sub.3
2-methyl-4-methoxy-phenyl --CH(CH.sub.2OR)(n-butyl) --CH.sub.3
2,4-dimethoxy-phenyl --CH(CH.sub.2OR)(n-butyl) --CH.sub.3
4-dimethylamino-2-methyl- --CH(CH.sub.2OR)(n-butyl) --CH.sub.3
3-pyridyl 4-dimethylamino-6-methyl- --CH(CH.sub.2OR)(n-butyl)
--CH.sub.3 3-pyridyl 4-dimethylamino-3-pyridyl
--CH(CH.sub.2OR)(n-butyl) --CH.sub.3 2,4-dichlorophenyl
--CH(CH.sub.2OR)(tert-butyl) --CH.sub.3 2-chloro-4-methyl-phenyl
--CH(CH.sub.2OR)(tert-butyl) --CH.sub.3 2-methyl-4-chloro-phenyl
--CH(CH.sub.2OR)(tert-butyl) --CH.sub.3 2,4,6-trimethyl-phenyl
--CH(CH.sub.2OR)(tert-butyl) --CH.sub.3 2-chloro-4-methoxy-phenyl
--CH(CH.sub.2OR)(tert-butyl) --CH.sub.3 2-methyl-4-methoxy-phenyl
--CH(CH.sub.2OR)(tert-butyl) --CH.sub.3 2,4-dimethoxy-phenyl
--CH(CH.sub.2OR)(tert-butyl) --CH.sub.3 4-dimethylamino-2-methyl-
--CH(CH.sub.2OR)(tert-butyl) --CH.sub.3 3-pyridyl
4-dimethylamino-6-methyl- --CH(CH.sub.2OR)(tert-butyl) --CH.sub.3
3-pyridyl 4-dimethylamino-3-pyridyl --CH(CH.sub.2OR)(tert-butyl)
--CH.sub.3 2,4-dichlorophenyl --CH(CH.sub.2OR)(4-chloro-benzyl)
--CH.sub.3 2-chloro-4-methyl-phenyl --CH(CH.sub.2OR)(4-chloro-benz-
yl) --CH.sub.3 2-methyl-4-chloro-phenyl
--CH(CH.sub.2OR)(4-chloro-b- enzyl) --CH.sub.3
2,4,6-trimethyl-phenyl --CH(CH.sub.2OR)(4-chloro-- benzyl)
--CH.sub.3 2-chloro-4-methoxy-phenyl
--CH(CH.sub.2OR)(4-chloro-benzyl) --CH.sub.3
2-methyl-4-methoxy-phenyl --CH(CH.sub.2OR)(4-chloro-benzyl)
--CH.sub.3 2,4-dimethoxy-phenyl --CH(CH.sub.2OR)(4-chloro-benzyl)
--CH.sub.3 4-dimethylamino-2-methyl-
--CH(CH.sub.2OR)(4-chloro-benzyl) --CH.sub.3 3-pyridyl
4-dimethylamino-6-methyl- --CH(CH.sub.2OR)(4-chlo- ro-benzyl)
--CH.sub.3 3-pyridyl 4-dimethylamino-3-pyridyl
--CH(CH.sub.2OR)(4-chloro-benzyl) --CH.sub.3 2,4-dichlorophenyl
--CH(CH.sub.2OR)(CH.sub.2CH.sub.2SCH.sub.3) --CH.sub.3
2-chloro-4-methyl-phenyl
--CH(CH.sub.2OR)(CH.sub.2CH.sub.2SCH.sub.3) --CH.sub.3
2-methyl-4-chloro-phenyl --CH(CH.sub.2OR)(CH.sub.2CH.su-
b.2SCH.sub.3) --CH.sub.3 2,4,6-trimethyl-phenyl
--CH(CH.sub.2OR)(CH.sub.2CH.sub.2SCH.sub.3) --CH.sub.3
2-chloro-4-methoxy-phenyl
--CH(CH.sub.2OR)(CH.sub.2CH.sub.2SCH.sub.3) --CH.sub.3
2-methyl-4-methoxy-phenyl --CH(CH.sub.2OR)(CH.sub.2CH.s-
ub.2SCH.sub.3) --CH.sub.3 2,4-dimethoxy-phenyl
--CH(CH.sub.2OR)(CH.sub.2CH.sub.2SCH.sub.3) --CH.sub.3
4-dimethylamino-2-methyl-
--CH(CH.sub.2OR)(CH.sub.2CH.sub.2SCH.sub.3) --CH.sub.3 3-pyridyl
4-dimethylamino-6-methyl-
--CH(CH.sub.2OR)(CH.sub.2CH.sub.2SCH.sub.3) --CH.sub.3 3-pyridyl
4-dimethylamino-3-pyridyl
--CH(CH.sub.2OR)(CH.sub.2CH.sub.2SCH.sub.3- ) --CH.sub.3
2,4-dichlorophenyl --CH(CH.sub.2OH.sub.3)(CH.sub.2Oben- zyl)
--CH.sub.3 2-chloro-4-methyl-phenyl --CH(CH.sub.2CH.sub.3)(CH.-
sub.2Obenzyl) --CH.sub.3 2-methyl-4-chloro-phenyl
--CH(CH.sub.2CH.sub.3)(CH.sub.2Obenzyl) --CH.sub.3
2,4,6-trimethyl-phenyl --CH(CH.sub.2CH.sub.3)(CH.sub.2Obenzyl)
--CH.sub.3 2-chloro-4-methoxy-phenyl
--CH(CH.sub.2CH.sub.3)(CH.sub.2Obenzyl) --CH.sub.3
2-methyl-4-methoxy-phenyl --CH(CH.sub.2CH.sub.3)(CH.sub- .2Obenzyl)
--CH.sub.3 2,4-dimethoxy-phenyl --CH(CH.sub.2CH.sub.3)(C-
H.sub.2Obenzyl) --CH.sub.3 4-dimethylamino-2-methyl-
--CH(CH.sub.2CH.sub.3)(CH.sub.2Obenzyl) --CH.sub.3 3-pyridyl
4-dimethylamino-6-methyl- --CH(CH.sub.2CH.sub.3)(CH.sub.2Obenzyl)
--CH.sub.3 3-pyridyl 4-dimethylamino-3-pyridyl
--CH(CH.sub.2CH.sub.3)(CH.sub.2Obenzyl) --CH.sub.3
2,4-dichlorophenyl --CH(n-butyl).sub.2 --CH.sub.3
4-isopropyl-phenyl --CH(n-propyl).sub.2 --CH.sub.3 4-chloro-phenyl
--CH(n-propyl).sub.2 --CH.sub.3 4-methoxy-phenyl
--CH(n-propyl).sub.2 --CH.sub.3 4-t-butyl-phenyl
--CH(n-propyl).sub.2 --CH.sub.3 2-benzofuranyl --CH(n-propyl).sub.2
--CH.sub.3 3,4-dimethoxy-phenyl --CH(n-propyl).sub.2 --CH.sub.3
2-chloro-phenyl --CH(n-propyl).sub.2 --CH.sub.3 2-benzothiophenyl
--CH(n-propyl).sub.2 --CH.sub.3 4-trifluoromethyl-phenyl
--CH(n-propyl).sub.2 --CH.sub.3 4-methylthio-phenyl
--CH(n-propyl).sub.2 --CH.sub.3 3-isopropyl-6-methoxy-phenyl
--CH(n-propyl).sub.2 --CH.sub.3 4-trifluoromethoxy-phenyl
--CH(n-propyl).sub.2 --CH.sub.3 3-trifluoromethyl-phenyl
--CH(n-propyl).sub.2 --CH.sub.3 dibenzofuranyl --CH(n-propyl).sub.2
--CH.sub.3 2,4-dichlorophenyl 3-methylcyclohexyl --CH.sub.3 NOTE:
Each occurrence of R in this Table is independently selected from a
C.sub.1-6alkyl, such as methyl or ethyl.
[0054] The compounds of the present invention may be prepared by
known organic synthesis techniques, including the methods described
in more detail in the Examples, and may generally be utilized as
the free base. Alternatively, the compounds of this invention may
be used in the form of acid addition salts. Acid addition salts of
the free base amino compounds of the present invention may be
prepared by methods well known in the art, and may be formed from
organic and inorganic acids. Suitable organic acids include maleic,
fumaric, benzoic, ascorbic, succinic, methanesulfonic, acetic,
oxalic, propionic, tartaric, salicylic, citric, gluconic, lactic,
mandelic, cinnamic, aspartic, stearic, palmitic, glycolic,
glutamic, and benzenesulfonic acids. Suitable inorganic acids
include hydrochloric, hydrobromic, sulfuric, phosphoric, and nitric
acids.
[0055] The effectiveness of a compound as a CRF receptor antagonist
may be determined by various assay methods. Suitable CRF
antagonists of this invention are capable of inhibiting the
specific binding of CRF to its receptor and antagonizing activities
associated with CRF. A compound of structure (I) may be assessed
for activity as a CRF antagonist by one or more generally accepted
assays for this purpose, including (but not limited to) the assays
disclosed by DeSouza et al. (J. Neuroscience 7:88, 1987) and
Battaglia et al. (Synapse 1:572, 1987). As mentioned above,
suitable CRF antagonists include compounds which demonstrate CRF
receptor affinity. CRF receptor affinity may be determined by
binding studies that measure the ability of a compound to inhibit
the binding of a radiolabeled CRF (e.g., [.sup.125I]tyrosine-CFR)
to its receptor (e.g., receptors prepared from rat cerebral cortex
membranes). The radioligand binding assay described by DeSouza et
al. (supra, 1987) provides an assay for determining a compound's
affinity for the CRF receptor. Such activity is typically
calculated from the IC.sub.50 as the concentration of a compound
necessary to displace 50% of the radiolabeled ligand from the
receptor, and is reported as a "K.sub.i" value calculated by the
following equation: 1 K i = IC 50 1 + L / K D
[0056] where L=radioligand and K.sub.D=affinity of radioligand for
receptor (Cheng and Prusoff, Biochem. Pharmacol. 22:3099,
1973).
[0057] In addition to inhibiting CRF receptor binding, a compound's
CRF receptor antagonist activity may be established by the ability
of the compound to antagonize an activity associated with CRF. For
example, CRF is known to stimulate various biochemical processes,
including adenylate cyclase activity. Therefore, compounds may be
evaluated as CRF antagonists by their ability to antagonize
CRF-stimulated adenylate cyclase activity by, for example,
measuring cAMP levels. The CRF-stimulated adenylate cyclase
activity assay described by Battaglia et al. (supra, 1987) provides
an assay for determining a compound's ability to antagonize CRF
activity. Accordingly, CRF receptor antagonist activity may be
determined by assay techniques which generally include an initial
binding assay (such as disclosed by DeSouza (supra, 1987)) followed
by a cAMP screening protocol (such as disclosed by Battaglia
(supra, 1987)).
[0058] With reference to CRF receptor binding affinities, CRF
receptor antagonists of this invention have a K.sub.i of less than
10 .mu.M. In a preferred embodiment of this invention, a CRF
receptor antagonist has a K.sub.i of less than 1 .mu.M, and more
preferably less than 0.25 .mu.M (i.e., 250 nM).
[0059] The CRF receptor antagonists of the present invention
demonstrate activity at the CRF receptor site, and may be used as
therapeutic agents for the treatment of a wide range of disorders
or illnesses including endocrine, psychiatric, and neurologic
disorders or illnesses. More specifically, the CRF receptor
antagonists of the present invention may be useful in treating
physiological conditions or disorders arising from the
hypersecretion of CRF. Because CRF is believed to be a pivotal
neurotransmitter that activates and coordinates the endocrine,
behavioral and automatic responses to stress, the CRF receptor
antagonists of the present invention can be used to treat
neuropsychiatric disorders. Neuropsychiatric disorders which may be
treatable by the CRF receptor antagonists of this invention include
affective disorders such as depression; anxiety-related disorders
such as generalized anxiety disorder, panic disorder,
obsessive-compulsive disorder, abnormal aggression, cardiovascular
abnormalities such as unstable angina and reactive hypertension;
and feeding disorders such as anorexia nervosa, bulimia, and
irritable bowel syndrome. CRF antagonists may also be useful in
treating stress-induced immune suppression associated with various
disease states, as well as stroke. Other uses of the CRF
antagonists of this invention include treatment of inflammatory
conditions (such as rheumatoid arthritis, uveitis, asthma,
inflammatory bowel disease and G.I. motility), Cushing's disease,
infantile spasms, epilepsy and other seizures in both infants and
adults, and various substance abuse and withdrawal (including
alcoholism).
[0060] In another embodiment, compounds of this invention and their
analogs may be used as Positron Emission Tomography (PET) ligands,
Single Photon Emission Computed Tomography (SPECT) ligands, or
other diagnostic radiopharmaceutical agents. Incorporation of an
appropriate isotope (such as .sup.11C or .sup.18F for PET or
.sup.125I in the case of SPECT) may provide an agent useful for the
diagnosis or therapeutic management of a patient. In addition, use
of a compound of the present invention may provide a physiological,
functional, or biological assessment of a patient or provide
disease or pathology detection and assessment.
[0061] In another embodiment of the invention, pharmaceutical
compositions containing one or more CRF receptor antagonists are
disclosed. For the purposes of administration, the compounds of the
present invention may be formulated as pharmaceutical compositions.
Pharmaceutical compositions of the present invention comprise a CRF
receptor antagonist of the present invention (i.e., a compound of
structure (I)) and a pharmaceutically acceptable carrier and/or
diluent. The CRF receptor antagonist is present in the composition
in an amount which is effective to treat a particular
disorder--that is, in an amount sufficient to achieve CRF receptor
antagonist activity, and preferably with acceptable toxicity to the
patient. Preferably, the pharmaceutical compositions of the present
invention may include a CRF receptor antagonist in an amount from
0.1 mg to 250 mg per dosage depending upon the route of
administration, and more preferably from 1 mg to 60 mg. Appropriate
concentrations and dosages can be readily determined by one skilled
in the art.
[0062] Pharmaceutically acceptable carrier and/or diluents are
familiar to those skilled in the art. For compositions formulated
as liquid solutions, acceptable carriers and/or diluents include
saline and sterile water, and may optionally include antioxidants,
buffers, bacteriostats and other common additives. The compositions
can also be formulated as pills, capsules, granules, or tablets
which contain, in addition to a CRF receptor antagonist, diluents,
dispersing and surface active agents, binders, and lubricants. One
skilled in this art may further formulate the CRF receptor
antagonist in an appropriate manner, and in accordance with
accepted practices, such as those disclosed in Remington's
Pharmaceutical Sciences, Gennaro, Ed., Mack Publishing Co., Easton,
Pa. 1990.
[0063] In another embodiment, the present invention provides a
method for treating a variety of disorders or illnesses, including
endocrine, psychiatric and neurologic disorders or illnesses. Such
methods include administering of a compound of the present
invention to a warm-blooded animal in an amount sufficient to treat
the disorder or illness. Such methods include systemic
administration of a CRF receptor antagonist of this invention,
preferably in the form of a pharmaceutical composition. As used
herein, systemic administration includes oral and parenteral
methods of administration. For oral administration, suitable
pharmaceutical compositions of CRF receptor antagonists include
powders, granules, pills, tablets, and capsules as well as liquids,
syrups, suspensions, and emulsions. These compositions may also
include flavorants, preservatives, suspending, thickening and
emulsifying agents, and other pharmaceutically acceptable
additives. For parental administration, the compounds of the
present invention can be prepared in aqueous injection solutions
which may contain, in addition to the CRF receptor antagonist,
buffers, antioxidants, bacteriostats, and other additives commonly
employed in such solutions.
[0064] As mentioned above, administration of a compound of the
present invention can be used to treat a wide variety of disorders
or illnesses. In particular, the compounds of the present invention
may be administered to a warm-blooded animal for the treatment of
depression, anxiety disorder, panic disorder, obsessive-compulsive
disorder, abnormal aggression, unstable angina, reactive
hypertension, anorexia nervosa, bulimia, irritable bowel syndrome,
stress-induced immune suppression, stroke, inflammation, Cushing's
disease, infantile spasms, epilepsy, and substance abuse or
withdrawal.
[0065] The following examples are provided for purposes of
illustration, not limitation.
EXAMPLES
[0066] The CRF receptor antagonists of this invention may be
prepared by the methods disclosed in Examples 1-2. Example 3
presents a method for determining the receptor binding activity
(K.sub.i) of compounds of this invention, while Example 4 discloses
an assay for screening compounds of this invention for
CRF-stimulated adenylate cyclase activity.
Example 1
Synthesis of Representative Compounds of Structure (Ia)
[0067] 89
Structure (Ia')
[0068] 3-amino-2-(2,4,6-trimethyl)phenyl Pyridine (2) (when X=N and
Ar=2,4,6-trimethylphenyl)
[0069] A mixture of 3-amino-2-chloropyridine (1) (2 g, 15 mmol, 1
eq.), trimethylphenylboronic acid ("Ar--B(OH).sub.2") (2.3 g, 14
mmol, 0.9 eq.), cesium fluoride (4.7 g, 31 mmol, 2 eq.) and
tetrakis(triphenylphosp- hine)palladium(0) (0.5 g, 4% mol) were
heated at reflux in anhydrous DME under N.sub.2 atmosphere
overnight. After cooling down at room temperature, solvents were
evaporated under reduced pressure. The residue was dissolved in
ethyl acetate, washed with water (3.times.50 mL). The aqueous
phases were combined, extracted with ethyl acetate (3.times.50 mL).
The organic phases were combined, extracted with a brine solution
(50 mL) and dried with Na.sub.2SO.sub.4. Compound (2) was purified
by liquid chromatography on silica gel with hexanes/ethyl acetate
8/2 as eluent mixture (Rf=0.4). 3 g of a transparent oil was
obtained. LC/MS (positive) 213 (M+1).
[0070] 3-amino-2-(2,4-dichloro)phenyl Pyridine (2') (when X=M and
Ar=2,4-dichlorophenyl)
[0071] The same procedure was employed as for compound (2) above,
but using 2,4-dichlorophenylboronic acid in place of
Ar--B(OH).sub.2. LC/MS (positive) 238 (M+1).
[0072] 4-hydroxy 2-methyl
8-(2,4,6-trimethyl)phenyl-1,7-naphthyridine (3)
[0073] A solution of 1.5 g of 3-amino-2-(2,4,6-trimethyl)phenyl
pyridine (2) (7.07 mmol, 1 eq.), 1.6 mL of ethylacetoacetate (12
mmol, 1.7 eq.) and a 200 mg of paratoluenesulfonic acid in 40 mL of
m-xylene was refluxed with a Dean-Stark trap for about 1.5 hour.
The m-xylene was removed. The residue was added to 4 mL of
diphenylether and heated until all the intermediate compound
reacted (about 10 min). The ring-closure was followed by LC/MS.
After cooling down at room temperature, 100 mL of hexanes were
added allowing compound (3) to crash out and used without further
purification. LC/MS (positive) 279 (M+1).
[0074] 4-hydroxy-2-methyl 8-(2,4-dichloro)phenyl-1,7-naphthyridine
(3')
[0075] Same procedure was employed as for compound (3) above, but
using compound (2') in place of compound (2). LC/MS positive 305
(M+1).
[0076] 4-chloro 2-methyl
8-(2,4,6-trimethyl)phenyl-1,7-naphthyridine (4)
[0077] 4-hydroxy-2-methyl-8-trimethylphenyl-1,7-naphthyridine (3)
was refluxed in 10 mL of POCl.sub.3 for 5 hours. After cooling down
at room temperature, the reaction mixture was poured on ice and
neutralized with a 6N NaOH solution. The product was extracted with
ethyl acetate (3.times.50 mL). The organic phases were combined,
washed with water (2.times.50 mL), a brine solution (1.times.50 mL)
and dried with sodium thiosulfate. Compound (4) was purified by
liquid chromatography on silica gel (hexanes/ethyl acetate 9/1, Rf
0.6). The reaction was quantitative. LC/MS (positive) 297
(M+1).
[0078] 4-chloro-2-methyl-8-(2,4-dichloro)phenyl-1,7-naphthyridine
(4')
[0079] Same procedure was employed as for compound (4) above, but
using compound (3') in place of compound (3). LC/MS (positive) 342
(M+1).
[0080]
4-chloro-2-methyl-5-nitro-8-(2,4,6-trimethyl)phenyl-17-naphthyridin-
e (5)
[0081] 4-chloro-2-methyl-8-trimethylphenyl-1,7-naphthyridine (4)
(521 mg, 1.76 mmol, 1 eq.) was added to 0.9 mL of sulfuric acid 97%
in an ice-bath. The mixture was heated at 55.degree. C. and 124
.mu.l of nitric acid 70% (1.93 mmol, 1.1 eq.) were added. The
reaction mixture was then stirred at 55 C. for 5 hours, cooled
down, poured into 20 mL of ice water and partially neutralized with
6N NaOH solution. The product was then extracted with ethyl acetate
(3.times.50 mL). The organic phases were combined, washed with
water (2.times.50 mL), a brine solution (1.times.50 mL) and dried
with sodium thiosulfate. Compound (5) was used with further
purification for the following step. LC/MS (positive) 342
(M+1).
[0082]
4-chloro-2-methyl-5-nitro-8-(2,4-dichloro)phenyl-1,7-naphthyridine
(5')
[0083] Same procedure was employed as for compound (5) above but
using compound (4') in place of compound (4).
[0084] 4-(N-4-heptyl
amino)-2-methyl-5-nitro-8-(2,4,6-trimethyl)phenyl-1.7-
-naphthyridine (6)
[0085]
4-chloro-2-methyl-5-nitro-8-(2,4,6-trimethyl)phenyl-1,7-naphthyridi-
ne (5) was heated in an excess of 4-heptylamine (0.5 mL) with
paratoluene sulfonic acid at 165.degree. C. in a reacti-vac
overnight. After cooling down at room temperature, the reaction
mixture was diluted in ethyl acetate and passed through a plug of
silica gel. Solvents were evaporated and compound (6) used in the
following step without purification. The reaction was quantitative.
LCV/MS (positive) 421 (M+1).
[0086] 4-(N-4-heptyl
Amino)-2-methyl-5-nitro-8-(2.4-dichloro)phenyl-1,7-na- phthyridine
(6')
[0087] Same procedure was employed as for compound (6) above, but
using compound (5') in place of compound (5).
[0088]
4-(N-4-heptylamino)-2-methyl-5-amino-8-(2,4,6-trimethyl)phenyl-1,7--
naphthyridine (7)
[0089]
4-(N-4-heptylamino)-2-methyl-5-nitro-8-(2,4,6-trimethyl)phenyl-1,7--
naphthyridine (6) was added to a suspension of palladium on carbon
(10% and a drop of acetic acid in methanol. The mixture was shaken
for 20 hours under hydrogen pressure (35 psi) at room temperature.
The catalyst was removed by filtration under celite and the
solvents evaporated. Compound (7) was used in the following step
without purification. The reaction was quantitative. LC/MS
(positive) 391 (M+1).
[0090]
4-(N-4-heptylamino)-2-methyl-5-amino-8-(2,4-dichloro)phenyl-1,7-nap-
hthyridine (7')
[0091] To a solution of
4-(N-4-heptylamino)-2-methyl-5-nitro-8-(2,4-dichlo-
ro)phenyl-1,7-naphthyridine (6') (153 mg, 0.36 mmol, 1 eq.) in THF
(8.5 mL) was added a solution of Na.sub.2--S.sub.2--O.sub.3 in
water (4.3 mL). The reaction mixture was heated at 60.degree. C.
for 2 hours. After cooling down at room temperature th4 product was
extracted with ethyl acetate (3.times.50 mL). The organic phases
were combined, washed with water (2.times.50 mL), a brine solution
(1.times.50 mL) and dried with sodium thiosulfate. Solvents were
evaporated and compound (7') was purified by PTLC (ethyl
acetate/hexanes 1/9 Rf 0.1).
[0092] Compound (8)
[0093] 100 mg of
4-(N-4-heptylamino)-2-methyl-5-amino-8-(2,4,6-trimethyl)
phenyl-1,7-naphthyridine (7) (0.4 mmol) was treated with
triethylorthoformate (2 mL) at reflux for 16 hours. After cooling
down at room temperature and evaporation of the solvent, the
product was extracted with ethyl acetate (3.times.20 mL). The
organic phases were combined, washed with water (10 mL), a brine
solution (10 mL) and dried with sodium thiosulfate. Solvents were
evaporated and compound (8) was purified by PTLC (ethyl
acetate/hexanes 1/1). LC/MS (positive) 400 (M+1).
[0094] Compound (8')
[0095] Same procedure was employed as for compound (8), but using
compound (7') in place of compound (7).
Structure (Ia")
[0096] Compounds of structure (Ia") may be made by the same
procedures as disclosed above, but employing 2-chloro-aniline as
compound (1) in place of 3-amino-2-chloropyridine. By this
technique, the compounds listed in the following Table 2 were
prepared.
2TABLE 2 REPRESENTATIVE COMPOUNDS OF STRUCTURE (IA") 10 Ar R.sub.1
R.sub.2 MW 2,4-dichlorophenyl --CH(n-propyl).sub.2 --CH.sub.3 426
4,6-dimethoxy-phenyl --CH(n-propyl).sub.2 --CH.sub.3 417
4-methoxy-6-methyl-phenyl --CH(n-propyl).sub.2 --CH.sub.3 401
2,4-dichlorophenyl --CH(ethyl).sub.2 --CH.sub.3 398
2,4-dichlorophenyl --CH(n-butyl).sub.2 --CH.sub.3 454
4-isopropyl-phenyl --CH(n-propyl).sub.2 --CH.sub.3 400 (M + 1)
4-chloro-phenyl --CH(n-propyl).sub.2 --CH.sub.3 393 (M + 1)
4-methoxy-phenyl --CH(n-propyl).sub.2 --CH.sub.3 388 (M + 1)
4-t-butyl-phenyl --CH(n-propyl).sub.2 --CH.sub.3 414 (M + 1)
2-benzofuranyl --CH(n-propyl).sub.2 --CH.sub.3 398 (M + 1)
3,4-dimethoxy-phenyl --CH(n-propyl).sub.2 --CH.sub.3 392
2-chloro-phenyl --CH(n-propyl).sub.2 --CH.sub.3 399
2-benzothiophenyl --CH(n-propyl).sub.2 --CH.sub.3 413
4-trifluoromethyl-phenyl --CH(n-propyl).sub.2 --CH.sub.3 425
4-methylthio-phenyl --CH(n-propyl).sub.2 --CH.sub.3 403
5-isopropyl-2-methoxy-phenyl --CH(n-propyl).sub.2 --CH.sub.3 429
4-trifluoromethoxy-phenyl --CH(n-propyl).sub.2 --CH.sub.3 441
3-trifluoromethyl-phenyl --CH(n-propyl).sub.2 --CH.sub.3 425
dibenzofuranyl --CH(n-propyl).sub.2 --CH.sub.3 447
2,4-dichlorophenyl 3-methylcyclohexyl --CH.sub.3 424
Example 2
Synthesis of Representative Compounds of Structure (Ib) and
(Ic)
[0097] 11
Structures (Ib') and (Ic')
[0098] Compound (9)
[0099] 100 mg of
4-(N-4-heptylamino)-2-methyl-5-amino-8-(2,4,6-trimethyl)
phenyl-1,7-naphthyridine (7) (0.4 mmol), 30 .mu.l of dibromopropane
and 100 mg of K.sub.2CO4 were dissolved in 2 mL of 2-butanone. The
reaction mixture was heated at 85.degree. C. for 4 hours in a
reacti-vac. After cooling down at room temperature compound (9) was
extracted with ethyl acetate (3.times.20 mL). The organic phases
were combined, washed with water (10 mL), a brine solution (10 mL)
and dried with sodium thiosulfate. Solvents were evaporated and
compound (9) was purified by PTLC (ethyl acetate/hexanes 1/1).
[0100] Compound (9')
[0101] Same procedure was employed as for compound (9), but using
compound (7') in place of compound (7).
[0102] Compound (10)
[0103] 100 mg of
4-(4-heptylamino)-2-methyl-5-amino-8-(2,4,6-trimethyl)
phenyl-1,7-naphthyridine (7) (0.4 mmol), 30 .mu.l of dibromopropane
and 100 mg of K.sub.2CO.sub.3 were dissolved in 2 mL of 2-butanone.
The reaction mixture was heated at 85.degree. C. for 4 hours in a
reacti-vac. After cooling down at room temperature compound (10)
was extracted with ethyl acetate (3.times.20 mL). The organic
phases were combined, washed with water (10 mL), a brine solution
(10 mL) and dried with sodium thiosulfate. Solvents were evaporated
and compound (10) was purified by PTLC (ethyl acetate/hexanes 1/1).
LC/MS (positive) 414 (M+1).
[0104] Compound (10')
[0105] Same procedure was employed as for compound (10), but using
compound (7') in place of compound (7).
Structures (Ib") and (Ic")
[0106] Compounds of structure (Ib") and (Ic") may be made by the
same procedures as disclosed above, but employing, 2-chloro-aniline
as compound (1) (X.dbd.CH) in place of 3-amino-2-chloropyridine. By
this technique the following representative compound of structure
(Ic") was made: Ar=2,4-dichlorophenyl, R.sub.1=--CH(n-propyl).sub.2
and R.sub.2=--CH.sub.3 (MW=442).
Example 3
Representative Compounds Having CRF Receptor Binding Activity
[0107] The compounds of this invention may be evaluated for binding
activity to the CRF receptor by a standard radioligand binding
assay as generally described by DeSouza et al. (J. Neurosci.
7:88-100, 1987). By utilizing various radiolabeled CRF ligands, the
assay may be used to evaluate the binding activity of the compounds
of the present invention with any CRF receptor subtype. Briefly,
the binding assay involves the displacement of a radiolabeled CRF
ligand from the CRF receptor.
[0108] More specifically, the binding assay is performed in 1.5 mL
Eppendorf tubes using approximately 1.times.10.sup.6 cells per tube
stably transfected with human CRF receptors. Each tube receives
about 0.1 mL of assay buffer (e.g., Dulbecco's phosphate buffered
saline, 10 mM magnesium chloride, 20 .mu.M bacitracin) with or
without unlabeled sauvagine, urotensin I or CRF (final
concentration, 1 .mu.M) to determine nonspecific binding, 0.1 mL of
[.sup.125I] tyrosine--ovine CRF (final concentration .about.200 pM
or approximately the K.sub.D as determined by Scatchard analysis)
and 0.1 mL of a membrane suspension of cells containing the CRF
receptor. The mixture is incubated for 2 hours at 22.degree. C.
followed by the separation of the bound and free radioligand by
centrifugation. Following two washes of the pellets, the tubes are
cut just above the pellet and monitored in a gamma counter for
radioactivity at approximately 80% efficiency. All radioligand
binding data may be analyzed using the non-linear least-square
curve-fitting program LIGAND of Munson and Rodbard (Anal. Biochem.
107:220, 1990).
Example 4
CRF-Stimulated Adenylate Cyclase Activity
[0109] The compounds of the present invention may also be evaluated
by various functional testing. For example, the compounds of the
present invention may be screened for CRF-stimulated adenylate
cyclase activity. An assay for the determination of CRF-stimulated
adenylate cyclase activity may be performed as generally described
by Battaglia et al. (Synapse 1:572, 1987), with modifications to
adapt the assay to whole cell preparations.
[0110] More specifically, the standard assay mixture may contain
the following in a final volume of 0.5 mL: 2 mM L-glutamine, 20 mM
HEPES, and 1 mM IMBX in DMEM buffer. In stimulation studies, whole
cells with the transfected CRF receptors are plated in 24-well
plates and incubated for 1 h at 37.degree. C. with various
concentrations of CRF-related and unrelated peptides in order to
establish the pharmacological rank-order profile of the particular
receptor subtype. Following the incubation, the media is aspirated,
the wells rinsed once gently with fresh media, and the media
aspirated. To determine the amount of intracellular cAMP, 300 .mu.l
of a solution of 95% ethanol and 20 mM aqueous hydrochloric acid is
added to each well and the resulting suspensions are incubated at
-20.degree. C. for 16 to 18 hours. The solution is removed into 1.5
mL Eppendorf tubes and the wells washed with an additional 200
.mu.l of ethanol/aqueous hydrochloric acid and pooled with the
first fraction. The samples are lyophilized and then resuspended
with 500 .mu.l sodium acetate buffer. The measurement of cAMP in
the samples is performed using a single antibody kit from
Biomedical Technologies Inc. (Stoughton, Mass.). For the functional
assessment of the compounds, a single concentration of CRF or
related peptides causing 80% stimulation of cAMP production is
incubated along with various concentrations of competing compounds
(10.sup.-12 to 10.sup.-6 M).
[0111] It will be appreciated that, although specific embodiments
of the invention have been described herein for purposes of
illustration, various modifications may be made without departing
from the spirit and scope of the invention. Accordingly, the
invention is not limited except as by the appended claims.
* * * * *