U.S. patent application number 11/749433 was filed with the patent office on 2007-09-20 for chemical compounds.
This patent application is currently assigned to Glaxo Group Limited. Invention is credited to Romano Di Fabio, Fabrizio Micheli, Yves St-Denis.
Application Number | 20070219232 11/749433 |
Document ID | / |
Family ID | 9918655 |
Filed Date | 2007-09-20 |
United States Patent
Application |
20070219232 |
Kind Code |
A1 |
Di Fabio; Romano ; et
al. |
September 20, 2007 |
Chemical Compounds
Abstract
The present invention provides compounds of formula (I)
including stereoisomers, prodrugs and pharmaceutically acceptable
salts or solvates thereof ##STR1## wherein R is aryl or heteroaryl,
each of which may be substituted by 1 to 4 groups selected from:
halogen, C1-C6 alkyl, C1-C6 alkoxy, halo C1-C6 alkyl, C2-C6
alkenyl, C2-C6 alkynyl, halo C1-C6 alkoxy, --C(O)R.sub.5, nitro,
--NR.sub.6R.sub.7, cyano, and a group R.sub.8; R.sub.1 is hydrogen,
C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo C1-C6 alkyl, halo
C1-C6 alkoxy, halogen, NR.sub.6R.sub.7 or cyano; R.sub.2 is
hydrogen, C3-C7 cycloalkyl, or a group R.sub.9; R.sub.3 is C3-C7
cycloalkyl, or a group R.sub.9; or R.sub.2 and R.sub.3 together
with N form a 5-14 membered heterocycle, which may be substituted
by 1 to 3 R.sub.10 groups; R.sub.4 is hydrogen, C1-C6 alkyl,
halogen or halo C1-C6 alkyl; R.sub.5 is a C1-C4 alkyl, --OR.sub.6
or --NR.sub.6R.sub.7; R.sub.6 is hydrogen or C1-C6 alkyl; R.sub.7
is hydrogen or C1-C6 alkyl; R.sub.8 is a 5-6 membered heterocycle,
which may be saturated or may contain one to three double bonds,
and which may be substituted by 1 or more R.sub.11 groups; R.sub.9
is a C1-C6 alkyl that may be substituted by one or more groups
selected from: C3-C7 cycloalkyl, C1-C6 alkoxy, haloC1-C6 alkoxy,
hydroxy, haloC1-C6 alkyl; R.sub.10 is a group R.sub.8, C3-C7
cycloalkyl, C1-C6 alkyl, C1-C6 alkoxy, halo C1-C6 alkyl, C2-C6
alkenyl, C2-C6 alkynyl, halo C1-C6 alkoxy, hydroxy, halogen, nitro,
cyano, C(O)NR.sub.6R.sub.7, phenyl which may be substituted by 1 to
4 R.sub.11 groups; R.sub.11 is C3-C7 cycloalkyl, C1-C6 alkyl, C1-C6
alkoxy, halo C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo C1-C6
alkoxy, hydroxy, halogen, nitro, cyano, or C(O)NR.sub.6R.sub.7; X
is carbon or nitrogen; n is 1 or 2; to processes for their
preparation, to pharmaceutical compositions containing them and to
their use in the treatment of conditions mediated by
corticotropin-releasing factor (CRF).
Inventors: |
Di Fabio; Romano; (Verona,
IT) ; Micheli; Fabrizio; (Verona, IT) ;
St-Denis; Yves; (Verona, IT) |
Correspondence
Address: |
GLAXOSMITHKLINE;Corporate Intellectual Property - UW2220
P.O. Box 1539
King of Prussia
PA
19406-0939
US
|
Assignee: |
Glaxo Group Limited
|
Family ID: |
9918655 |
Appl. No.: |
11/749433 |
Filed: |
May 16, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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10483792 |
Jan 14, 2004 |
7253284 |
|
|
PCT/EP02/07865 |
Jul 15, 2002 |
|
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|
11749433 |
May 16, 2007 |
|
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Current U.S.
Class: |
514/300 ;
546/113; 546/193 |
Current CPC
Class: |
A61P 25/24 20180101;
A61P 1/08 20180101; A61P 25/28 20180101; A61P 5/38 20180101; A61P
11/02 20180101; A61P 1/04 20180101; A61P 1/14 20180101; A61P 17/00
20180101; A61P 11/00 20180101; A61P 17/06 20180101; A61P 9/10
20180101; A61P 25/22 20180101; C07D 471/04 20130101; A61P 43/00
20180101; A61P 9/00 20180101; A61P 25/00 20180101; A61P 3/04
20180101; A61P 11/16 20180101; A61P 1/00 20180101; A61P 37/08
20180101; A61P 25/20 20180101; A61P 29/00 20180101 |
Class at
Publication: |
514/300 ;
546/113; 546/193 |
International
Class: |
A61K 31/44 20060101
A61K031/44; C07D 211/00 20060101 C07D211/00; C07D 471/00 20060101
C07D471/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 17, 2001 |
GB |
0117396.2 |
Claims
1. Compounds of formula (I) including stereoisomers, prodrugs and
pharmaceutically acceptable salts or solvates thereof ##STR21## R
is aryl or heteroaryl, each of which may be substituted by 1 to 4
groups selected from: halogen, C1-C6 alkyl, C1-C6 alkoxy, halo
C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo C1-C6 alkoxy,
--C(O)R.sub.5, nitro, --NR.sub.6R.sub.7, cyano, and a group
R.sub.8; R.sub.1 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6
alkynyl, halo C1-C6 alkyl, halo C1-C6 alkoxy, halogen,
NR.sub.6R.sub.7 or cyano; R.sub.2 is hydrogen, C3-C7 cycloalkyl, or
a group R.sub.9; R.sub.3 is C3-C7 cycloalkyl, or a group R.sub.9;
or R.sub.2 and R.sub.3 together with N form a 5-14 membered
heterocycle, which may be substituted by 1 to 3 R.sub.10 groups;
R.sub.4 is hydrogen, C1-C6 alkyl, halogen or halo C1-C6 alkyl;
R.sub.5 is a C1-C4 alkyl, --OR.sub.6 or --NR.sub.6R.sub.7; R.sub.6
is hydrogen or C1-C6 alkyl; R.sub.7 is hydrogen or C1-C6 alkyl;
R.sub.8 is a 5-6 membered heterocycle, which may be saturated or
may contain one to three double bonds, and which may be substituted
by 1 or more R.sub.11 groups; R.sub.0 is a C1-C6 alkyl that may be
substituted by one or more groups selected from: C3-C7 cycloalkyl,
C1-C6 alkoxy, haloC1-C6 alkoxy, hydroxy, haloC1-C6 alkyl; R.sub.10
is a group R.sub.8, C3-C7 cycloalkyl, C1-C6 alkyl, C1-C6 alkoxy,
halo C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo C1-C6 alkoxy,
hydroxy, halogen, nitro, cyano, C(O)NR.sub.6R.sub.7, phenyl which
may be substituted by 1 to 4 R.sub.11 groups; R.sub.11 is C3-C7
cycloalkyl, C1-C6 alkyl, C1-C6 alkoxy, halo C1-C6 alkyl, C2-C6
alkenyl, C2-C6 alkynyl, halo C1-C6 alkoxy, hydroxy, halogen, nitro,
cyano, or C(O)NR.sub.6R.sub.7; X is carbon or nitrogen; n is 1 or
2.
2. Compounds, according to claim 1, in which R.sub.2 and R.sub.3
together with N form a 5-14 membered heterocyclic group, which may
be substituted by 1 to 3 R.sub.10 groups.
3. Compounds, according to claim 1, of general formula (Ia)
##STR22## in which R, R.sub.1, R.sub.2, R.sub.3 R.sub.4 and X are
defined as in claim 1.
4. Compounds, according to claim 3, of general formula (IIa)
##STR23## in which R, R.sub.1, R.sub.2, R.sub.4 and R.sub.3 are
defined as in claim 1.
5. Compounds, according to claim 4, in which the group
NR.sub.2R.sub.3 represents a 5-6 membered heterocyclic group, which
may be substituted by 1 to 3 R.sub.8 groups.
6. Compounds, according to claim 5, of general formula (IIIa)
##STR24## wherein R, R.sub.1, R.sub.4 and R.sub.9 are defined as in
claim 1.
7. Compounds, according to claim 1, of general formula (Ib)
##STR25## in which R, R.sub.1, R.sub.2, R.sub.3 R.sub.4 and X are
defined as in claim 1.
8. Compounds, according to claim 7, of general formula (IIIb)
##STR26## in which R, R.sub.1, R.sub.2, R.sub.4 and R.sub.3 are
defined as in claim 1.
9. Compounds, according to claim 1 wherein R.sub.1 is C1-C3 alkyl
group or halo C1-C3 alkyl group and R.sub.4 is hydrogen.
10. Compounds, according claim 1 wherein R is an aryl group
selected from: 2,4-dichlorophenyl, 2-chloro-4-methylphenyl,
2-chloro-4-trifluoromethyl, 2-chloro-4-methoxyphenyl,
2,4,5-trimethylphenyl, 2,4-dimethylphenyl,
2-methyl-4-methoxyphenyl, 2-methyl4-chlorophenyl,
2-methyl-4-trifluoromethyl, 2,4-dimethoxyphenyl,
2-methoxy-4-trifluoromethylphenyl, 2-methoxy-4-chlorophenyl,
3-methoxy-4-chlorophenyl, 2,5-dimethoxy-4-chlorophenyl,
2-methoxy-4-isopropylphenyl, 2-methoxy-4-trifluoromethylphenyl,
2-methoxy-4-isopropylphenyl, 2-methoxy-4-methylphenyl,
2-trifluoromethyl-4-chlorophenyl, 2,4-trifluoromethylphenyl,
2-trifluoromethyl-4-methylphenyl,
2-trifluoromethyl-4-methoxyphenyl, 2-bromo-4-isopropylphenyl,
2-methyl4-cyanophenyl, 2-chloro-4-cyanophenyl,
4-methyl-6-dimethylaminopyridin-3-yl, 3,5-dichloro-pyridin-2-yl,
2,6-bismethoxy-pyridin-3-yl and
3-chloro-5-tricloromethyl-pyridin-2-yl.
11. A compound, according claim 1 selected in a group consisting
from:
1-(2,4-bis-trifluoromethyl-phenyl)-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-y-
l)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine;
1-(2,4-bis-trifluoromethyl-phenyl)-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-y-
l)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine;
3-methyl-4-[6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-2,3-dihydro-1H-pyrro-
lo[2,3-b]pyridin-1-yl]-benzonitrile;
4-[6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]-
pyridin-1-yl]-3-trifluoromethyl-benzonitrile;
6-methyl-1-(2-methyl-4-trifluoromethoxy-phenyl)-4-(3-thiazol-2-yl-pyrazol-
-1-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine;
1-(2,4-bis-trifluoromethyl-phenyl)-7-methyl-5-(3-thiazol-2-yl-pyrazol-1-y-
l)-1,2,3,4-tetrahydro-[1,8]naphthyridine;
1-(4-methoxy-2-methyl-phenyl)-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-2,-
3-dihydro-1H-pyrrolo[2,3-b]pyridine
1-(2,4-bis-trifluoromethyl-phenyl)-6-methyl-4-(3-morpholin-4-yl-pyrazol-1-
-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine
1-(2,4-bis-trifluoromethyl-phenyl)-6-methyl-4-(3-pyridin-2-yl-pyrazol-1-y-
l)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine
4-[1,3']bipyrazolyl-1'-yl-1-(2,4-bis-trifluoromethyl-phenyl)-6-methyl-2,3-
-dihydro-1H-pyrrolo[2,3-b]pyridine.
12. A pharmaceutical composition comprising a compound according to
claim 1 in admixture with one or more physiologically acceptable
carriers or excipients.
13. A method for the treatment of a mammal, including man, in
particular in the treatment of conditions mediated by CRF
(corticotropin-releasing factor), comprising administration of an
effective amount of a compound according to claim 1.
14. A method, according to claim 13, in the treatment of depression
and anxiety, comprising administration of an effective amount of a
compound according to claim 1.
15. A method, according to claim 14, in the treatment of IBS
(irritable bowel disease) and IBD (inflammatory bowel disease),
comprising administration of an effective amount of a compound
according to claim 1.
Description
[0001] This application is a continuation of application Ser. No.
10/483,792, filed Jan. 14, 2004, now pending, which is a 371 of
International Application No. PCT/EP02/07865, filed Jul. 15,
2002.
[0002] The present invention relates to bicyclic derivatives, to
processes for their preparation, to pharmaceutical compositions
containing them and to their use in therapy.
[0003] The first corticotropin-releasing factor (CRF) was isolated
from ovine hypothalami and identified as a 41-amino acid peptide
(Vale et al., Science 213: 1394-1397,1981). 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"), Bendorphin and other proopiomelanocortin
("POMC")-derived peptides from the anterior pituitary (Vale et al.,
Science 213: 1394-1397,1981).
[0004] In addition to its role in stimulating the production of
ACTH and POMC, 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.
[0005] 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.
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.
[0006] 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.
[0007] WO 95/10506 describes inter alia compounds of general
formula (A) with general CRF antagonist activity ##STR2##
[0008] wherein Y may be CR29; V may be nitrogen, Z may be carbon,
R3 may correspond to an amine derivative and R4 may be taken
together with R29 to form a 5-membered ring and is --CH(R28) when
R29 is-CH(R30). There are no specific disclosures of compounds
corresponding to this definition.
[0009] WO 95/33750 also describes compounds of general formula (B)
having CRF antagonistic activity, ##STR3##
[0010] in which A and Y may be nitrogen and carbon and B may
correspond to an amine derivative. There are no specific
disclosures of compounds corresponding to this definition.
[0011] WO 98/08846 describes compounds of general formula (C)
having CRF antagonistic activity, ##STR4##
[0012] wherein A may be carbon, G may be nitrogen or carbon, B may
be an amino derivative and the other groups have the meanings as
defined.
[0013] 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.
[0014] 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.
[0015] In particular the invention relates to novel compounds which
are potent and specific antagonists of corticotropin-releasing
factor (CRF) receptors.
[0016] The present invention provides compounds of formula (I)
including stereoisomers, prodrugs and pharmaceutically acceptable
salts or solvates thereof ##STR5## [0017] wherein [0018] R is aryl
or heteroaryl, each of which may be substituted by 1 to 4 groups
selected from: [0019] halogen, C1-C6 alkyl, C1-C6 alkoxy, halo
C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo C1-C6 alkoxy,
--C(O)R.sub.5, nitro, --NR.sub.6R.sub.7, cyano, and a group
R.sub.8; [0020] R.sub.1 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl,
C2-C6 alkynyl, halo C1-C6 alkyl, halo C1-C6 alkoxy, halogen,
NR.sub.6R.sub.7 or cyano; [0021] R.sub.2 is hydrogen, C3-C7
cycloalkyl, or a group R.sub.9; [0022] R.sub.3 is C3-C7 cycloalkyl,
or a group R.sub.9; or [0023] R.sub.2 and R.sub.3 together with N
form a 5-14 membered heterocycle, which may be substituted by 1 to
3 R.sub.10 groups; [0024] R.sub.4 is hydrogen, C1-C6 alkyl, halogen
or halo C1-C6 alkyl; [0025] R.sub.5 is a C1-C4 alkyl, --OR.sub.6 or
--NR.sub.6R.sub.7; [0026] R.sub.6 is hydrogen or C1-C6 alkyl;
[0027] R.sub.7 is hydrogen or C1-C6 alkyl; [0028] R.sub.8 is a 5-6
membered heterocycle, which may be saturated or may contain one to
three double bonds, and which may be substituted by 1 or more
R.sub.11 groups; [0029] R.sub.9 is a C1-C6 alkyl that may be
substituted by one or more groups selected from: C3-C7 cycloalkyl,
C1-C6 alkoxy, haloC1-C6 alkoxy, hydroxy, haloC1-C6 alkyl; [0030]
R.sub.10 is a group R.sub.8, C3-C7 cycloalkyl, C1-C6 alkyl, C1-C6
alkoxy, halo C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halo C1-C6
alkoxy, hydroxy, halogen, nitro, cyano, C(O)NR.sub.6R.sub.7, phenyl
which may be substituted by 1 to 4 R.sub.11 groups; [0031] R.sub.11
is C3-C7 cycloalkyl, C1-C6 alkyl, C1-C6 alkoxy, halo C1-C6 alkyl,
C2-C6 alkenyl, C2-C6 alkynyl, halo C1-C6 alkoxy, hydroxy, halogen,
nitro, cyano, or C(O)NR.sub.6R.sub.7; [0032] X is carbon or
nitrogen; [0033] n is 1 or 2.
[0034] 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, malic, fumaric, benzoic, ascorbic,
succinic, methanesulfonic, p-toluensulfonic, 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.
Thus, the term "pharmaceutically acceptable salt" of structure (I)
is intended to encompass any and all acceptable salt forms.
[0035] The solvates may, for example, be hydrates.
[0036] References hereinafter to a compound according to the
invention include both compounds of formula (I) and their
pharmaceutically acceptable acid addition salts together with
pharmaceutically acceptable solvates.
[0037] In addition, prodrugs are also included within the context
of this invention. Prodrugs are any covalently bonded carriers that
release a compound of structure (I) in vivo when such prodrug is
administered to a patient. Prodrugs are generally prepared by
modifying functional groups in a way such that the modification is
cleaved, either by routine manipulation or in vivo, yielding the
parent compound. Prodrugs include, for example, compounds of this
invention wherein hydroxy, amine or sulfhydryl groups are bonded to
any group that, when administered to a patient, cleaves to form the
hydroxy, amine or sulfhydryl groups. Thus, representative examples
of prodrugs include (but are not limited to) acetate, formate and
benzoate derivatives of alcohol, sulfhydryl and amine functional
groups of the compounds of structure (I). Further, in the case of a
carboxylic acid (--COOH), esters may be employed, such as methyl
esters, ethyl esters, and the like.
[0038] With regard to stereoisomers, the compounds of structure (I)
may have chiral centers and may occur as recemates, racemic
mixtures and as individual enantiomers or diastereomers. All such
isomeric forms are included within the present invention, including
mixtures thereof. Furthermore, some of the crystalline forms of the
compounds of structure (I) may exist as polymorphs, which are
included in the present invention.
[0039] The term C1-C6 alkyl as used herein as a group or a part of
the group refers to a linear or branched alkyl group containing
from 1 to 6 carbon atoms; examples of such groups include methyl,
ethyl, propyl, isopropyl, n-butyl, isobutyl, tert butyl, pentyl or
hexyl.
[0040] The term C3-C7 cycloalkyl group means a non aromatic
monocyclic hydrocarbon ring of 3 to 7 carbon atom such as, for
example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or
cycloheptyl; while unsaturated cycloalkyls include cyclopentenyl
and cyclohexenyl, and the like.
[0041] The term halogen refers to a fluorine, chlorine, bromine or
iodine atom.
[0042] The term halo C1-C6 alkyl, or halo C1-C2 alkyl means an
alkyl group having one or more carbon atoms and wherein at least
one hydrogen atom is replaced with halogen such as for example a
trifluoromethyl group and the like.
[0043] The term C2-C6 alkenyl defines straight or branched chain
hydrocarbon radicals containing one or more double bond and having
from 2 to 6 carbon atoms such as, for example, ethenyl, 2-propenyl,
3-butenyl, 2-butenyl, 2-pentenyl, 3-pentenyl, 3-methyl-2-butenyl or
3-hexenyl and the like.
[0044] The term C1-C6 alkoxy group may be a linear or a branched
chain alkoxy group, for example methoxy, ethoxy, propoxy,
prop-2-oxy, butoxy, but-2-oxy or methylprop-2-oxy and the like.
[0045] The term halo C1-C6 alkoxy group may be a C1-C6 alkoxy group
as defined before substituted with at least one halogen, preferably
fluorine, such as OCHF.sub.2, or OCF.sub.3.
[0046] The term C2-C6 alkynyl defines straight or branched chain
hydrocarbon radicals containing one or more triple bond and having
from 2 to 6 carbon atoms including acetylenyl, propynyl, 1-butynyl,
1-pentynyl, 3-methyl-1-butynyl and the like.
[0047] The term aryl means an aromatic carbocyclic moiety such as
phenyl, biphenyl or naphthyl.
[0048] The term heteroaryl means an aromatic heterocycle ring of 5
to 10 members and having at least one heteroatom selected from
nitrogen, oxygen and sulfur, and containing at least 1 carbon atom,
including both mono-and bicyclic ring systems.
[0049] Representative heteroaryls include (but are not limited to)
furyl, benzofuranyl, thiophenyl, benzothiophenyl, pyrrolyl,
indolyl, isoindolyl, azaindolyl, pyridyl, quinolinyl,
isoquinolinyl, oxazolyl, isooxazolyl, benzoxazolyl, pyrazolyl,
imidazolyl, benzimidazolyl, thiazolyl, benzothiazolyl,
isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl,
cinnolinyl, phthalazinyl, triazolyl, tetrazolyl, and
quinazolinyl.
[0050] The term 5-14 membered heterocycle means a 5 to 7-membered
monocyclic, or 7-to 14-membered polycyclic, heterocycle ring which
is either saturated, unsaturated or aromatic, and which contains
from 1 to 4 heteroatoms independently selected from nitrogen,
oxygen and sulfur, and wherein the nitrogen and sulfur heteroatoms
may be optionally oxidized, and the nitrogen heteroatom may be
optionally quaternized, including bicyclic rings in which any of
the above heterocycles are fused to a benzene ring as well as
tricyclic (and higher) heterocyclic rings. The heterocycle may be
attached via any heteroatom or carbon atom. Heterocycles include
heteroaryls as defined above. Thus, in addition to the aromatic
heteroaryls listed above, heterocycles also include (but are not
limited to) morpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl,
hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl,
tetrahydropyranyl, tetrahydropyridinyl, tetrahydropyrimidinyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydropyrimidinyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.
[0051] The term 5-6 membered heterocycle means, according to the
above definition, a 5-6 monocyclic heterocyclic ring which is
either saturated, unsaturated or aromatic, and which contains from
1 to 4 heteroatoms independently selected from nitrogen, oxygen and
sulfur, and wherein the nitrogen and sulfur heteroatoms may be
optionally oxidized, and the nitrogen heteroatom may be optionally
quaternized. Heterocycles include heteroaryls as defined above. The
heterocycle may be attached via any heteroatom or carbon atom.
Thus, the term include (but are not limited to) morpholinyl,
pyridinyl, pyrazinyl, pyrazolyl, thiazolyl, triazolyl, imidazolyl,
oxadiazolyl, oxazolyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl,
hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl,
tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydropyrimidinyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.
[0052] Representative compounds of this invention include the
following structure (Ia) and (Ib) ##STR6##
[0053] In one preferred embodiment in which n is 1, according to
the definition of the compounds of formula (I) above, the CRF
receptor antagonists of this invention have structure (Ia), and,
when n is 2, then the CRF receptor antagonists of this invention
have structure (Ib), wherein R, R.sub.1, R.sub.2 and R.sub.3 are
defined as above.
[0054] Further representative compounds of this invention include
compounds of general formula (I) in which [0055] R.sub.2 and
R.sub.3 together with N form a 5-14 membered heterocyclic group,
which may be substituted by 1 to 3 R.sub.10 groups; such R.sub.10
groups are defined as above.
[0056] Depending upon the choice of X, the CRF receptor antagonists
of this invention include compounds of formula (IIa) and (IIb), in
which the group NR.sub.2R.sub.3 represents a 5-6 membered
heterocyclic group, which may be substituted by 1 to 3 R.sub.8
groups. ##STR7##
[0057] In particular compounds of formula (IIa) and (IIb) are
included ##STR8##
[0058] in which R.sub.1, R and R.sub.8 have the meanings as defined
before.
[0059] Examples of such compounds are reported in the Experimental
Part.
[0060] Even more preferred embodiments of the invention include,
but are not limited to, compounds of the formula (I), (Ia), (Ib),
(IIa), (IIb), (IIIa), (IIIb):
[0061] wherein: [0062] R.sub.1 is C1-C3 alkyl group or halo C1-C3
alkyl group, preferably methyl or trifluoromethyl; [0063] R.sub.4
is hydrogen; and [0064] R is an aryl group selected from:
2,4-dichlorophenyl, 2-chloro-4-methylphenyl,
2-chloro-4-trifluoromethyl, 2-chloro-4-methoxyphenyl,
2,4,5-trimethylphenyl, 2,4-dimethylphenyl,
2-methyl-4-methoxyphenyl, 2-methyl-4-chlorophenyl,
2-methyl-4-trifluoromethyl, 2,4-dimethoxyphenyl,
2-methoxy-4-trifluoromethylphenyl, 2-methoxy-4-chlorophenyl,
3-methoxy-4-chlorophenyl, 2,5-dimethoxy-4-chlorophenyl,
2-methoxy-4-isopropylphenyl, 2-methoxy-4-trifluoromethylphenyl,
2-methoxy-4-isopropylphenyl, 2-methoxy-4-methylphenyl,
2-trifluoromethyl-4-chlorophenyl, 2,4-trifluoromethylphenyl,
2-trifluoromethyl-4-methylphenyl,
2-trifluoromethyl-4-methoxyphenyl, 2-bromo4-isopropylphenyl,
2-methyl-4-cyanophenyl, 2-chloro-4-cyanophenyl,
4-methyl-6-dimethylaminopyridin-3-yl, 3,5-dichloro-pyridin-2-yl,
2,6-bismethoxy-pyridin-3-yl and
3-chloro-5-trichloromethyl-pyridin-2-yl.
[0065] Preferred compounds according to the invention are:
[0066]
1-(2,4-bis-trifluoromethyl-phenyl)-6-methyl-4-(3-thiazol-2-yl-pyra-
zol-1-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine;
[0067]
1-(2,4-bis-trifluoromethyl-phenyl)-6-methyl-4-(3-thiazol-2-yl-pyra-
zol-1-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine;
[0068]
3-methyl-4-[6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-2,3-dihydro-1-
H-pyrrolo[2,3-b]pyridin-1-yl]-benzonitrile;
[0069]
4-[6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-2,3-dihydro-1H-pyrrolo-
[2,3-b]pyridin-1-yl]-3-trifluoromethyl-benzonitrile;
[0070]
6-methyl-1-(2-methyl-4-trifluoromethoxy-phenyl)-4-(3-thiazol-2-yl--
pyrazol-1-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine;
[0071]
1-(2,4-bis-trifluoromethyl-phenyl)-7-methyl-5-(3-thiazol-2-yl-pyra-
zol-1-yl)-1,2,3,4-bismethoxy-pyridin-1,8]naphthyridine;
[0072]
1-(4-methoxy-2-methyl-phenyl)-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-
-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine
[0073]
1-(2,4-bis-trifluoromethyl-phenyl)-6-methyl-4-(3-morpholin-4-yl-py-
razol-1-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine
[0074]
1-(2,4-bis-trifluoromethyl-phenyl)-6-methyl-4-(3-pyridin-2-yl-pyra-
zol-1-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine
[0075]
4-[1,3']bipyrazolyl-1'-yl-1-(2,4-bis-trifluoromethyl-phenyl)-6-met-
hyl-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine.
[0076] In general, the compounds of structure (I) may be made
according to the organic synthesis techniques known to those
skilled in this field, as well as by the representative methods set
forth in the Examples.
[0077] Compounds of formula (I), and salts and solvates thereof,
may be prepared by the general methods outlined hereinafter. In the
following description, the groups R, R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8, R.sub.9, R.sub.10,
R.sub.11, and n have the meanings as previously defined for
compounds of formula (I) unless otherwise stated.
[0078] Compounds of formula (IIa) may be conveniently prepared
according to the following Scheme 1: ##STR9##
[0079] in which [0080] step a stands for conversion of the leaving
group L, selected in a group consisting from: halogen or reactive
residue of sulphonic acid (e.g. mesylate, tosylate), preferably
chloride, in the amino group of compounds (III), by reaction with
the suitable amine NR.sub.2R.sub.3 in basic conditions; [0081] step
b stands for reduction of the ester group with a suitable reducing
agent (such as DIBAl--H) to hydroxy group of compounds (IV); [0082]
step c stands for oxidation of the hydroxy group with a suitable
oxidising agent (such as Dess-Martin periodinane) to aldehyde group
of compound (V); [0083] step d stands for formation of the aldehyde
group of compounds (VII) by Wittig reaction in the usual
conditions, through formation of enol ether followed by acid
hydrolysis (step e); [0084] step f stands for reduction of the
aldehyde group with a suitable reducing agent (such as NaBH.sub.4)
to hydroxy group of compounds (VIII); [0085] step g stands for
conversion of the hydroxy group in the suitable protecting group of
compounds (IX)(such as TBS: tert-butyldimethylsilyl); [0086] step h
stands for Buchwald reaction by coupling with the suitable amine
RNH.sub.2; [0087] step i stands for deprotection reaction to give
the hydroxy group of compounds (XI); [0088] step l stands for
intramolecular cyclisation by heating after conversion of the
hydroxy group of compounds (XI) in a suitable leaving group (such
as bromide, by reaction with CBr.sub.4 and PPh.sub.3) to give the
final compounds (IIa).
[0089] Alternatively, compounds of formula (IIa) may be
conveniently prepared according to the following Scheme 2:
##STR10##
[0090] in which [0091] step a' stands for conversion of the hydroxy
group in a suitable leaving group L' of compounds (XII), which,
independently from L, has the same definition (e.g mesylate, by
reaction with MsCl in Et.sub.3N); [0092] step b' stands for
conversion of L' in the cyano derivative of compounds (XIII) by
reaction with, e.g. KCN in an aprotic dipolar solvent, like DMF;
[0093] step c' stands for reduction of the cyano group with a
suitable reducing agent agent (such as BH.sub.3-THF) to the amino
group of compound (XIV); [0094] step d' stands for intramolecular
cyclisation of compounds (XIV) by heating in a suitable solvent
(such as NMP) at high temperature; [0095] step e' stands for
reduction of the aldehyde group with a suitable reducing agent
(such as NaBH.sub.4) to hydroxy group of compounds (VIII); [0096]
step f' corresponds to previous step h.
[0097] Compounds of formula (IIb) may be conveniently prepared
according to the following Scheme 3: ##STR11##
[0098] in which: [0099] step a'' corresponds to previous step d;
[0100] step b'' corresponds to previous step e; [0101] step c''
corresponds to previous step f; [0102] step d'' corresponds to
previous step g; [0103] step e'' corresponds to previous step h;
[0104] step f'' corresponds to previous step i; [0105] step g''
corresponds to previous step l;
[0106] Compounds of formula (II) are known compounds or may be
prepared according to known methods in the literature.
[0107] Compounds of formula (IIIa) and (IIIb) may be prepared
according to the previous Schemes 1, 2 and 3, once prepared the
heterocyclic reactive residue according to known methods to the
skilled in the art.
[0108] Examples of suitable hydroxy protecting group include
trihydrocarbyl silyl ethers such as the trimethylsilyl or
t-butyldimethylsilyl ether. The hydroxyl protecting groups may be
removed by well-known standard procedures (such as those described
in Protective Groups in Organic Chemistry, pages 46-119, Edited by
J F W McOmie (Plenum Press, 1973)). For example when Pg is a
t-butyldimethylsilyl group, this may be removed by treatment with
triethylamine trihydrofluoride.
[0109] Pharmaceutical acceptable salts may also be prepared from
other salts, including other pharmaceutically acceptable salts, of
the compound of formula (I) using conventional methods.
[0110] The compounds of formula (I) may readily be isolated in
association with solvent molecules by crystallisation or
evaporation of an appropriate solvent to give the corresponding
solvates.
[0111] When a specific enantiomer of a compound of general formula
(I) is required, this may be obtained for example by resolution of
a corresponding enantiomeric mixture of a compound of formula (I)
using conventional methods. Thus the required enantiomer may be
obtained from the racemic compound of formula (I) by use of chiral
HPLC procedure.
[0112] The subject invention also includes isotopically-labeled
compounds, which are identical to those recited in formulas I and
following, but for the fact that one or more atoms are replaced by
an atom having an atomic mass or mass number different from the
atomic mass or mass number usually found in nature. Examples of
isotopes that can be incorporated into compounds of the invention
include isotopes of hydrogen, carbon, nitrogen, oxygen,
phosphorous, fluorine, iodine, and chlorine, such as .sup.3H,
.sup.11C, .sup.14C, .sup.18F, .sup.123I and .sup.125I. Compounds of
the present invention and pharmaceutically acceptable salts of said
compounds that contain the aforementioned isotopes and/or other
isotopes of other atoms are within the scope of the present
invention. Isotopically-labeled compounds of the present invention,
for example those into which radioactive isotopes such as .sup.3H,
.sup.14C are incorporated, are useful in drug and/or substrate
tissue distribution assays. Tritiated, i.e., .sup.3H, and
carbon-14, i.e., .sup.14C, isotopes are particularly preferred for
their ease of preparation and detectability. .sup.11C and .sup.8F
isotopes are particularly useful in PET (positron emission
tomography), and .sup.125I isotopes are particularly useful in
SPECT (single photon emission computerized tomography), all useful
in brain imaging. Further, substitution with heavier isotopes such
as deuterium, i.e., .sup.2H, can afford certain therapeutic
advantages resulting from greater metabolic stability, for example
increased in vivo half-life or reduced dosage requirements and,
hence, may be preferred in some circumstances. Isotopically labeled
compounds of formula I and following of this invention can
generally be prepared by carrying out the procedures disclosed in
the Schemes and/or in the Examples below, by substituting a readily
available isotopically labeled reagent for a non-isotopically
labeled reagent.
[0113] The CRF receptor antagonists of the present invention
demonstrate activity at the CRF receptor site including CRF 1 and
CRF 2 receptors and may be used in the treatment of conditions
mediated by CRF or CRF receptors.
[0114] 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).
[0115] The CRF receptors-binding assay was performed by using the
homogeneous technique of scintillation proximity (SPA). The ligand
binds to recombinant membrane preparation expressing the CRF
receptors which in turn bind to wheatgerm agglutinin coated SPA
beads. In the Experimental Part will be disclosed the details of
the experiments.
[0116] With reference to CRF receptor binding affinities, CRF
receptor antagonists of this invention have a Ki less than 10
.mu.m.
[0117] Compounds of the invention are useful in the treatment of
central nervous system disorders where CRF receptors are involved.
In particular in the treatment or prevention of major depressive
disorders including bipolar depression, unipolar depression, single
or recurrent major depressive episodes with or without psychotic
features, catatonic features, melancholic features, atypical
features or postpartum onset, the treatment of anxiety and the
treatment of panic disorders. Other mood disorders encompassed
within the term major depressive disorders include dysthymic
disorder with early or late onset and with or without atypical
features, neurotic depression, post traumatic stress disorders and
social phobia; dementia of the Alzheimer's type, with early or late
onset, with depressed mood; vascular dementia with depressed mood;
mood disorders induced by alcohol, amphetamines, cocaine,
hallucinogens, inhalants, opioids, phencyclidine, sedatives,
hypnotics, anxiolytics and other substances; schizoaffective
disorder of the depressed type; and adjustment disorder with
depressed mood. Major depressive disorders may also result from a
general medical condition including, but not limited to, myocardial
infarction, diabetes, miscarriage or abortion, etc.
[0118] Compounds of the invention are also useful in the treatment
or prevention of schizophrenic disorders including paranoid
schizophrenia, disorganised schizophrenia, catatonic schizophrenia,
undifferentiated schizophrenia, residual schizoprenia.
[0119] Compounds of the invention are useful as analgesics. In
particular they are useful in the treatment of traumatic pain such
as postoperative pain; traumatic avulsion pain such as brachial
plexus; chronic pain such as arthritic pain such as occurring in
osteo-, rheumatoid or psoriatic arthritis; neuropathic pain such as
post-herpetic neuralgia, trigeminal neuralgia, segmental or
intercostal neuralgia, fibromyalgia, causalgia, peripheral
neuropathy, diabetic neuropathy, chemotherapy-induced neuropathy,
AIDS related neuropathy, occipital neuralgia, geniculate neuralgia,
glossopharyngeal neuralgia, reflex sympathetic dystrophy, phantom
limb pain; various forms of headache such as migraine, acute or
chronic tension headache, temporomandibular pain, maxillary sinus
pain, cluster headache; odontalgia; cancer pain; pain of visceral
origin; gastrointestinal pain; nerve entrapment pain; sport's
injury pain; dysmennorrhoea; menstrual pain; meningitis;
arachnoiditis; musculoskeletal pain; low back pain e.g. spinal
stenosis; prolapsed disc; sciatica; angina; ankylosing
spondyolitis; gout; burns; scar pain; itch; and thalamic pain such
as post stroke thalamic pain.
[0120] Compounds of the invention are also useful for the treatment
of dysfunction of appetite and food intake and in circumstances
such as anorexia, anorexia nervosa and bulimia.
[0121] Compounds of the invention are also useful in the treatment
of sleep disorders including dysomnia, insomnia, sleep apnea,
narcolepsy, and circadian ritmic disorders.
[0122] Compounds of the invention are also useful in the treatment
or prevention of cognitive disorders. Cognitive disorders include
dementia, amnestic disorders and cognitive disorders not otherwise
specified.
[0123] Furthermore compounds of the invention are also useful as
memory and/or cognition enhancers in healthy humans with no
cognitive and/or memory deficit.
[0124] Compounds of the invention are also useful in the treatment
of tolerance to and dependence on a number of substances. For
example, they are useful in the treatment of dependence on
nicotine, alcohol, caffeine, phencyclidine (phencyclidine like
compounds), or in the treatment of tolerance to and dependence on
opiates (e.g. cannabis, heroin, morphine) or benzodiazepines; in
the treatment of cocaine, sedative ipnotic, amphetamine or
amphetamine-related drugs (e.g. dextroamphetamine,
methylamphetamine) addiction or a combination thereof.
[0125] Compounds of the invention are also useful as
anti-inflammatory agents. In particular they are useful in the
treatment of inflammation in asthma, influenza, chronic bronchitis
and rheumatoid arthritis; in the treatment of inflammatory diseases
of the gastrointestinal tract such as Crohn's disease, ulcerative
colitis, inflammatory bowel disease (IBD) and non-steroidal
anti-inflammatory drug induced damage; inflammatory diseases of the
skin such as herpes and eczema; inflammatory diseases of the
bladder such as cystitis and urge incontinence; and eye and dental
inflammation.
[0126] Compounds of the invention are also useful in the treatment
of allergic disorders, in particular allergic disorders of the skin
such as urticaria, and allergic disorders of the airways such as
rhinitis.
[0127] Compounds of the invention are also useful in the treatment
of emesis, i.e. nausea, retching and vomiting. Emesis includes
acute emesis, delayed emesis and anticipatory emesis. The compounds
of the invention are useful in the treatment of emesis however
induced. For example, emesis may be induced by drugs such as cancer
chemotherapeutic agents such as alkylating agents, e.g.
cyclophosphamide, carmustine, lomustine and chlorambucil; cytotoxic
antibiotics, e.g. dactinomycin, doxorubicin, mitomycin-C and
bleomycin; anti-metabolites, e.g. cytarabine, methotrexate and
5-fluorouracil; vinca alkaloids, e.g. etoposide, vinblastine and
vincristine; and others such as cisplatin, dacarbazine,
procarbazine and hydroxyurea; and combinations thereof; radiation
sickness; radiation therapy, e.g. irradiation of the thorax or
abdomen, such as in the treatment of cancer; poisons; toxins such
as toxins caused by metabolic disorders or by infection, e.g.
gastritis, or released during bacterial or viral gastrointestinal
infection; pregnancy; vestibular disorders, such as motion
sickness, vertigo, dizziness and Meniere's disease; post-operative
sickness; gastrointestinal obstruction; reduced gastrointestinal
motility; visceral pain, e.g. myocardial infarction or peritonitis;
migraine; increased intercranial pressure; decreased intercranial
pressure (e.g. altitude sickness); opioid analgesics, such as
morphine; and gastro-oesophageal reflux disease, acid indigestion,
over-indulgence of food or drink, acid stomach, sour stomach,
waterbrash/regurgitation, heartburn, such as episodic heartburn,
nocturnal heartburn, and meal-induced heartburn and dyspepsia.
[0128] Compounds of the invention are of particular use in the
treatment of gastrointestinal disorders such as irritable bowel
syndrome (IBS); skin disorders such as psoriasis, pruritis and
sunburn; vasospastic diseases such as angina, vascular headache and
Reynaud's disease; cerebral ischeamia such as cerebral vasospasm
following subarachnoid haemorrhage; fibrosing and collagen diseases
such as scleroderma and eosinophilic fascioliasis; disorders
related to immune enhancement or suppression such as systemic lupus
erythematosus and rheumatic diseases such as fibrositis; and
cough.
[0129] Compounds of the invention are useful for the treatment of
neurotoxic injury which follows cerebral stroke, thromboembolic
stroke, hemorrhagic stroke, cerebral ischemia, cerebral vasospam,
hypoglycemia, hypoxia, anoxia, perinatal asphyxia cardiac
arrest.
[0130] The invention therefore provides a compound of formula (I)
or a pharmaceutically acceptable salt or solvate thereof for use in
therapy, in particular in human medicine.
[0131] There is also provided as a further aspect of the invention
the use of a compound of formula (I) or a pharmaceutically
acceptable salt or solvate thereof in the preparation of a
medicament for use in the treatment of conditions mediated by
CRF.
[0132] In an alternative or further aspect there is provided a
method for the treatment of a mammal, including man, in particular
in the treatment of condition mediated by CRF, comprising
administration of an effective amount of a compound of formula (I)
or a pharmaceutically acceptable salt or a solvate thereof.
[0133] It will be appreciated that reference to treatment is
intended to include prophylaxis as well as the alleviation of
established symptoms.
[0134] Compounds of formula (I) may be administered as the raw
chemical but the active ingredient is preferably presented as a
pharmaceutical formulation.
[0135] Accordingly, the invention also provides a pharmaceutical
composition which comprises at least one compound of formula (I) or
a pharmaceutically acceptable salt thereof and formulated for
administration by any convenient route. Such compositions are
preferably in a form adapted for use in medicine, in particular
human medicine, and can conveniently be formulated in a
conventional manner using one or more pharmaceutically acceptable
carriers or excipients.
[0136] Thus compounds of formula (I) may be formulated for oral,
buccal, parenteral, topical (including ophthalmic and nasal), depot
or rectal administration or in a form suitable for administration
by inhalation or insufflation (either through the mouth or
nose).
[0137] For oral administration, the pharmaceutical compositions may
take the form of, for example, tablets or capsules prepared by
conventional means with pharmaceutically acceptable excipients such
as binding agents (e.g. pregelatinised maize starch,
polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers
(e.g. lactose, microcrystalline cellulose or calcium hydrogen
phosphate); lubricants (e.g. magnesium stearate, talc or silica);
disintegrants (e.g. potato starch or sodium starch glycollate); or
wetting agents (e.g. sodium lauryl sulphate). The tablets may be
coated by methods well known in the art. Liquid preparations for
oral administration may take the form of, for example, solutions,
syrups or suspensions, or they may be presented as a dry product
for constitution with water or other suitable vehicle before use.
Such liquid preparations may be prepared by conventional means with
pharmaceutically acceptable additives such as suspending agents
(e.g. sorbitol syrup, cellulose derivatives or hydrogenated edible
fats); emulsifying agents (e.g. lecithin or acacia); non-aqueous
vehicles (e.g. almond oil, oily esters, ethyl alcohol or
fractionated vegetable oils); and preservatives (e.g. methyl or
propyl-p-hydroxybenzoates or sorbic acid). The preparations may
also contain buffer salts, flavouring, colouring and sweetening
agents as appropriate.
[0138] Preparations for oral administration may be suitably
formulated to give controlled release of the active compound.
[0139] For buccal administration the composition may take the form
of tablets or formulated in conventional manner.
[0140] The compounds of the invention may be formulated for
parenteral administration by bolus injection or continuous
infusion. Formulations for injection may be presented in unit
dosage form e.g. in ampoules or in multi-dose containers, with an
added preservative. The compositions may take such forms as
suspensions, solutions or emulsions in oily or aqueous vehicles,
and may contain formulatory agents such as suspending, stabilising
and/or dispersing agents. Alternatively, the active ingredient may
be in powder form for constitution with a suitable vehicle, e.g.
sterile pyrogen-free water, before use.
[0141] The compounds of the invention may be formulated for topical
administration in the form of ointments, creams, gels, lotions,
pessaries, aerosols or drops (e.g. eye, ear or nose drops).
Ointments and creams may, for example, be formulated with an
aqueous or oily base with the addition of suitable thickening
and/or gelling agents. Ointments for administration to the eye may
be manufactured in a sterile manner using sterilised
components.
[0142] Lotions may be formulated with an aqueous or oily base and
will in general also contain one or more emulsifying agents,
stabilising agents, dispersing agents, suspending agents,
thickening agents, or colouring agents. Drops may be formulated
with an aqueous or non-aqueous base also comprising one or more
dispersing agents, stabilising agents, solubilising agents or
suspending agents. They may also contain a preservative.
[0143] The compounds of the invention may also be formulated in
rectal compositions such as suppositories or retention enemas, e.g.
containing conventional suppository bases such as cocoa butter or
other glycerides.
[0144] The compounds of the invention may also be formulated as
depot preparations. Such long acting formulations may be
administered by implantation (for example subcutaneously or
intramuscularly) or by intramuscular injection. Thus, for example,
the compounds of the invention may be formulated with suitable
polymeric or hydrophobic materials (for example as an emulsion in
an acceptable oil) or ion exchange resins, or as sparingly soluble
derivatives, for example, as a sparingly soluble salt.
[0145] For intranasal administration, the compounds of the
invention may be formulated as solutions for administration via a
suitable metered or unitary dose device or alternatively as a
powder mix with a suitable carrier for administration using a
suitable delivery device.
[0146] A proposed dose of the compounds of the invention is 1 to
about 1000 mg per day. It will be appreciated that it may be
necessary to make routine variations to the dosage, depending on
the age and condition of the patient and the precise dosage will be
ultimately at the discretion of the attendant physician or
veterinarian. The dosage will also depend on the route of
administration and the particular compound selected.
[0147] Thus for parenteral administration a daily dose will
typically be in the range of 1 to about 100 mg, preferably 1 to 80
mg per day. For oral administration a daily dose will typically be
within the range 1 to 300 mg e.g. 1 to 100 mg.
EXAMPLES
[0148] In the Intermediates and Examples unless otherwise
stated:
[0149] Melting points (m.p.) were determined on a Gallenkamp m.p.
apparatus and are uncorrected. All temperatures refers to .degree.
C. Infrared spectra were measured on a FT-IR instrument. Proton
Magnetic Resonance (.sup.1H-NMR) spectra were recorded at 400 MHz,
chemical shifts are reported in ppm downfield (d) from Me.sub.4Si,
used as internal standard, and are assigned as singlets (s),
doublets (d), doublets of doublets (dd), triplets (t), quartets (q)
or multiplets (m). Column chromathography was carried out over
silica gel (Merck AG Darmstaadt, Germany). The following
abbreviations are used in text: EtOAc=ethyl acetate,
cHex=cyclohexane, CH.sub.2Cl.sub.2=dichloromethane,
Et.sub.2O=dietyl ether, DMF=N,N'-dimethylformamide,
DIPEA=N,N-diisopropylethylamine, DME=ethylene glycol dimethyl
ether, MeOH=methanol, Et.sub.3N=triethylamine, TFA=trifluoroacetic
acid, THF=tetrahydrofuran, DIBAL-H=diisobutylaluminium hydride,
DMAP=dimethylaminopyridine, LHMDS=lithiumhexamethyldisilazane; Tlc
refers to thin layer chromatography on silica plates, and dried
refers to a solution dried over anhydrous sodium sulphate; r.t.
(RT) refers to room temperature.
[0150] Intermediate 1
2,4-Dichloro-6-methyl-nicotinic acid ethyl ester
[0151] The title compound was prepared according to an already
published procedure: Mittelbach, Martin; Synthesis, 1988, 6, p.
479-80.
[0152] Intermediate 2
2-Chloro-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-nicotinic acid
ethyl ester
[0153] To a solution of 2-(1H-pyrazol-3-yl)-1,3-thiazole (7.71 g,
1.05 eq) in anh. DMF (61 mL), at 0.degree. C., under N.sub.2, was
added NaH 60% in mineral oil (2.03 g, 1.05 eq) and the reaction
mixture was stirred for 10 min. at 0.degree. C. and then for 1 hr
at room temperature. Intermediate 1 (11.34 g, 48.0 mmol) was then
added as a solution in anh. DMF (35 mL) at 0.degree. C. and the
resulting solution was heated at 110.degree. C. for 3 hr. The
reaction was then quenched with water, extracted with EtOAc, washed
with brine, dried over anh. Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude product was purified by flash
chromatography (silica gel, cHex/EtOAc 7:3) to give 7.02 g of the
title compound as a white solid.
[0154] NMR (.sup.1H, CDCl.sub.3): .delta. 7.91 (d, 1H), 7.91 (d,
1H), 7.41 (d, 1H), 7.31 (s, 1H), 7.18 (d, 1H), 4.50 (q, 2H), 2.78
(s, 3H), 1.25 (t, 3H).
[0155] MS (m/z): 349 [MH].sup.+.
[0156] Intermediate 3
[2-Chloro-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-3-yl]-methanol
[0157] To a solution of intermediate 2 (1.5 g, 4.3 mmol) in anh.
CH.sub.2Cl.sub.2 (30 mL), at -78.degree. C., under N.sub.2, was
added DIBAl--H 1.0 M in cyclohexane (12.9 mL, 3.0 eq). The reaction
mixture was stirred for 1 hr at -78.degree. C. and then for 1 hr at
room temperature. The reaction was then quenched with a saturated
solution of Rochelle's salt, extracted with EtOAc, washed with
brine, dried over anh. Na.sub.2SO.sub.4, filtered and concentrated
in vacuo. The crude product was purified by flash chromatography
(silica gel, cHex/EtOAc 1:1) to give 1.02 g of the title compound
as a white solid.
[0158] NMR (.sup.1H, CDCl.sub.3): .delta. 8.05 (d, 1H), 7.90 (d,
1H), 7.40 (d, 1H), 7.25 (s, 1H), 7.10 (d, 1H), 4.65 (S, 2H), 4.0
(bs, 1H), 2.60 (s, 3H).
[0159] MS (m/z): 307 [M].sup.+.
[0160] Intermediate 4
2-Chloro-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridine-3-carbaldehyde
[0161] To a solution of intermediate 3 (150 mg, 0.5 mmol) in anh.
CH.sub.2Cl.sub.2 (5 mL), at room temperature, under N.sub.2, was
added the Dess Martin periodinane (237 mg, 1.12 eq) and the
reaction mixture was stirred for 1 hr at room temperature. The
reaction was then quenched with a solution of 0.5 g of sodium
thiosulfate dissolved in a saturated solution of sodium
bicarbonate, extracted with EtOAc, washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
product was purified by flash chromatography (silica gel,
cHex/EtOAc 1:1) to give 124 mg of the title compound as a white
solid.
[0162] NMR (.sup.1H, CDCl.sub.3): .delta. 10.4 (s, 1H), 8.0-7.9
(2d, 2H), 7.40 (2d, 2H), 7.10 (s, 1H), 2.70 (s, 3H).
[0163] MS (m/z): 305 [MH].sup.+.
[0164] Intermediate 5
2-Chloro-3-(2-methoxy-vinyl)-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyri-
dine
[0165] To a solution of (methoxymethyl)-triphenylphosphonium
chloride (4.24 g, 3 eq) in anh. THF (20 mL), at 0.degree. C., under
N.sub.2, was added n-BuLi 1.6 M in cyclohexane (7.73 ml, 12.37
mmol) and the reaction mixture was brought to room temperature and
then stirred for 15 min. A solution of intermediate 4 (1.25 g, 4.1
mmol) in anh. THF (15 mL) was then added and the reaction was
stirred at room temperature for 1.5 hr. The reaction was then
quenched with water, extracted with EtOAc, washed with brine, dried
over anh. Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude product was purified by flash chromatography (silica gel,
cHex/EtOAc 4:1) to give 961 mg of the title compound as a white
solid (E:Z=3:2 mixture, used as such in the next step).
[0166] NMR (.sup.1H, CDCl.sub.3) principal E product: .delta. 7.90
(m, 1H), 7.83 (m, 1H), 7.38 (m, 1H), 7.05 (m, 1H), 7.00 (m, 1H),
6.51 (d, 1H), 5.63 (d, 1H), 3.64 (s, 3H), 2.60 (s, 3H).
[0167] MS (m/z): 333 [MH].sup.+.
[0168] Intermediate 6
[2-Chloro-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-3-yl]-acetaldeh-
yde
[0169] To a solution of intermediate 5 (936 mg, 2.8 mmol) in anh.
THF (15 mL) was added 6N HCl (21 ml, 45 eq) and the reaction
mixture was stirred at room temperature for 15 hr. The reaction was
then quenched with sat. aq. NaHCO.sub.3 until pH=7, extracted with
EtOAc, washed with brine, dried over anh. Na.sub.2SO.sub.4,
filtered and concentrated in vacuo to give 893 mg of the title
compound as a white solid, which was used in the next step without
further purification.
[0170] NMR (.sup.1H, CDCl.sub.3): .delta. 9.80 (s, 1H), 7.90-7.80
(2d, 2H), 7.70 (d, 1H), 7.20 (d, 1H), 7.0 (s, 1H), 4.25 (s, 2H),
2.70 (s, 3H).
[0171] MS (m/z): 319 [MH].sup.+.
[0172] Intermediate 7
2-[2-Chloro-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-3-yl]-ethanol
[0173] To a solution of intermediate 6 (903 mg, 2.84 mmol) in anh.
MeOH (10 mL) were added CeCl.sub.3 (700 mg, 1 eq) and NaBH.sub.4
(107 mg, 1 eq) and the reaction mixture was stirred at room
temperature for 5 min. The reaction was then quenched with water,
extracted with ethyl acetate, washed with brine, dried over anh.
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give 848 mg
of the title compound as a white solid, which was used in the next
step without further purification.
[0174] NMR (.sup.1H, CDCl.sub.3): .delta. 8.00 (m, 2H), 7.50 (d,
1H), 7.20 (m, 2H), 4.25 (t, 2H), 3.20 (t, 2H), 2.70 (s, 3H).
[0175] MS (m/z): 321 [MH].sup.+.
[0176] Intermediate 8
3-[2-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-2-chloro-6-methyl-4-(3-thiazo-
l-2-yl-pyrazol-1-yl)-pyridine
[0177] To a solution of intermediate 7 (840 mg, 2.6 mmol) in anh.
CH.sub.2Cl.sub.2 (10 mL) were added 2,6-lutidine (0.67 ml, 2.2 eq)
and tert-butyldimethylsilyl triflate (0.89 ml, 1.5 eq) and the
reaction mixture was stirred at room temperature for 15 hr. The
reaction was then quenched with an aqueous solution of saturated
NH.sub.4Cl, extracted with EtOAc, washed with brine, dried over
anh. Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
product was purified by flash chromatography (silica gel,
cHex/EtOAc 3:2) to give 950 mg of the title compound as a colorless
oil.
[0178] NMR (.sup.1H, CDCl.sub.3): .delta. 8.20 (d, 1H), 7.75 (d,
1H), 7.35 (d, 1H), 7.00 (m, 2H), 4.00 (t, 2H), 3.05 (t, 2H), 2.55
(s, 3H), 0.80 (s, 9H), -0.10 (s, 6H).
[0179] MS (m/z): 435 [MH].sup.+.
[0180] Intermediate 9
(2,4-Bis-trifluoromethyl-phenyl)-[3-[2-(tert-butyl-dimethyl-silanyloxy)-et-
hyl]-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-2-yl]-amine
[0181] To a solution of intermediate 8 (240 mg, 0.553 mmol) in anh.
DME (1 mL) were added Pd.sub.2(dba).sub.3 (51 mg, 0.1 eq),
2-(dicyclohexylphosphino)-2'-methylbiphenyl (60 mg, 0.3 eq),
K.sub.3PO.sub.4 (317 mg, 3 eq) and 2,4-bis(trifluoromethyl) aniline
(0.17 ml, 2 eq) and the reaction mixture was submitted to microwave
irradiation (150 W, 100.degree. C., 60 psi) for 20 min. The
reaction was then quenched with an aqueous solution of saturated
NH.sub.4Cl, extracted with EtOAc, washed with brine, dried over
anh. Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
product was purified by flash chromatography (silica gel,
cHex/EtOAc 9:1) to give 180 mg of the title compound as a colorless
oil.
[0182] NMR (.sup.1CDCl.sub.3): .delta. 8.55 (d, 1H), 8.20 (bs, 1H),
7.90 (d, 1H), 7.80 (m, 2H), 7.65 (dd, 1H), 7.40 (d, 1H), 7.05 (d,
1H), 6.85 (s, 1H), 4.20 (t, 2H), 2.90 (t, 2H), 2.60 (s, 3H), 0.80
(s, 9H), 0.10 (s, 6H).
[0183] MS (m/z): 628 [MH].sup.+.
[0184] Intermediate 10
2-[2-(2,4-Bis-trifluoromethyl-phenylamino)-6-methyl-4-(3-thiazol-2-yl-pyra-
zol-1-yl)-pyridin-3-yl]-ethanol
[0185] To a solution of intermediate 9 (240 mg, 0.38 mmol) in anh.
THF (5 mL) was added Et.sub.3N.3HF (0.187 ml, 3 eq) and the
reaction mixture was stirred for 15 hr at room temperature. The
reaction was then quenched with an aqueous solution of saturated
NH.sub.4Cl, extracted with EtOAc, washed with brine, dried over
anh. Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
product was purified by flash chromatography (silica gel,
cHex/EtOAc 1:1) to give 180 mg of the title compound as a colorless
oil.
[0186] NMR (.sup.1H, CDCl.sub.3): .delta. 8.45 (bs, 1H), 8.20 (d,
1H), 7.85 (d, 1H), 7.85 (2d, 2H), 7.65 (dd, 1H), 7.30 (d, 1H), 7.05
(d, 1H), 6.85 (s, 1H), 4.20 (t, 2H), 2.85 (t, 2H), 2.50 (s,
3H).
[0187] MS (m/z): 514 [MH].sup.+.
[0188] Intermediate 11
Methanesulfonic acid
2-chloro-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-3-ylmethyl
ester
[0189] To a solution of intermediate 3 (308 mg, 1.01 mmol) in anh.
CH.sub.2Cl.sub.2 (2.5 mL), at -25.degree. C., under N.sub.2, was
added Et.sub.3N (280 .mu.L, 2 eq) and CH.sub.3SO.sub.2Cl (120
.mu.L, 1.5 eq). The reaction mixture was stirred at -25.degree. C.
for 2 hr and than at -5.degree. C. for another 2 hr. The reaction
mixture was diluted with water and extracted with CH.sub.2Cl.sub.2.
The combined organic extracts were dried over anh.
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
product was purified by flash chromatography (silica gel,
cHex/EtOAc 6:4.fwdarw.1:1) to give 88 mg of the title compound as a
colourless oil.
[0190] NMR (.sup.1H, CDCl.sub.3): .delta. 7.90 (d, 1H), 7.87 (d,
1H), 7.39 (d, 1H), 7.34 (s, 1H), 7.14 (d, 1H), 5.5 (s, 2H), 3.0 (s,
3H), 2.78 (s, 3H).
[0191] MS (m/z): 385 [MH].sup.+, Cl
[0192] Intermediate 12
[2-Chloro-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-3-yl]-acetonitr-
ile
[0193] To a solution of intermediate 11 (88 mg, 0.229 mmol) in anh.
DMF (2.5 mL), at 0.degree. C., under N.sub.2, was added KCN (15 mg,
1 eq). The reaction mixture was stirred at room temperature for 5
hr. The reaction mixture was diluted with water and 1M NaOH and
extracted with Et.sub.2O. The combined organic extracts were dried
over anh. Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
title compound was obtained as a yellow solid (60 mg) and was used
in the next step without further purification.
[0194] NMR (.sup.1H, CDCl.sub.3): .delta. 7.92 (d, 1H), 7.91 (d,
1H), 7.41 (d, 1H), 7.31 (s, 1H), 7.18 (d, 1H), 3.99 (s, 2H), 2.78
(s, 3H).
[0195] MS (m/z): 316 [MH].sup.+, Cl
[0196] Intermediate 13
2-[2-Chloro-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-3-yl]-ethylam-
ine
[0197] To a solution of intermediate 12 (810 mg, 2.571 mmol) in
anh. THF (6 mL), at r.t., under N.sub.2, was added BH.sub.3.THF
(10.3 mL, 4 eq). The reaction mixture was stirred at reflux
temperature for 2 hr. The reaction mixture was concentrated in
vacuo and diluted with MeOH. 1M HCl in Et.sub.2O (7.7 .mu.L, 3 eq)
was added at r.t. and the solution was stirred at reflux for 2 hr.
The reaction mixture was diluted with water and basified with 1M
NaOH to pH=12. The crude product was purified by flash
chromatography (silica gel CH.sub.2Cl.sub.2/MeOH 6:4). The title
compound was obtained as a pale yellow solid (690 mg).
[0198] NMR (.sup.1H, CDCl.sub.3): .delta. 8.42 (d, 1H), 7.94 (d,
1H), 7.79 (d, 1H), 7.48 (s, 1H), 7.07(d, 1H), 2.81 (m, 4H), 2.51
(s, 3H), 2.0 (bs, 2H).
[0199] MS (m/z): 320 [MH].sup.+, Cl
[0200] Intermediate 14
6-Methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyri-
dine
[0201] To a solution of intermediate 13 (640 mg, 2.01 mmol) in dry
N-methylpyrrolidinone (13 mL), at r.t., under N.sub.2, was added
Et.sub.3N (1.12 mL, 4 eq). The reaction mixture was stirred at
110.degree. C. for 7 hr. The reaction mixture was diluted with
water and extracted with EtOAc. The combined organic extracts were
dried over anh. Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The crude product was purified by flash chromatography
(silica gel, CH.sub.2Cl.sub.2/MeOH 98:2). The title compound was
obtained as white solid (187 mg).
[0202] NMR (.sup.1H, CDCl.sub.3): .delta. 7.98 (d, 1H), 7.89 (d,
1H), 7.35 (d, 1H), 7.06 (d, 1H), 4.65 (bs, 1H), 3.72 (t, 2H), 3.48
(t, 2H), 2.42 (s, 3H)
[0203] MS (m/z): 284 [MH].sup.+
[0204] Intermediate 15
2-Chloro-3-(3-methoxy-allyl)-6-methyl-4-(3-thiazol-2-1-pyrazol-1-yl)-pyrid-
ine
[0205] To a stirred suspension of
(methoxy-methyl)triphenylphosphonium chloride (833 mg, 3eq.) in
anh. THF (4 mL) was added dropwise, at 0.degree. C., under N.sub.2,
n-BuLi in hexanes 1.6 M (1.50 ml, 3 eq). The reaction mixture was
stirred at r.t. for 15 rnin before a solution of intermediate 6
(258 mg, I eq) in anh. THF (3 ml) was added. The reaction mixture
was stirred for 1.5 hr. The mixture was quenched with water and
extracted with EtOAc. The combined organic extracts were dried over
anh. Na.sub.2SO.sub.4, filtered and concentrated to dryness in
vacuo. The crude product was purified by flash chromatography
(silica gel, cHex/EtOAc 8:2) to give 220 mg of the title compound
as an unseparable mixture of trans and cis (60/40) vinyl ether
(yellow oil, 78%)
[0206] NMR (.sup.1H, CDCl.sub.3): .delta. 7.88 (d, 1H), 7.79 (d,
1H), 7.36 (d, 1H), 7.19 (s, 1H), 7.08 (d, 1H), 6.31 (d, 1H), 4.90
(m, 1H), 3.44 (d, 2H), 3.48 (s, 3H), 2.57 (s, 3H).
[0207] MS (m/z): 347 [MH].sup.+, 1 Cl
[0208] Intermediate 16
3-[2-Chloro-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-3-yl]-propion-
aldehyde
[0209] To a solution of intermediate 15 (370 mg, 1.07 mmol) in THF
(5 mL) was added HCl 6 N (12 ml, 67.5 eq) and the reaction mixture
was stirred for 13 hr at r.t. A solution of NaHCO.sub.3 was added
to the reaction mixture until pH=7 was reached and the aqueous
phase was extracted with EtOAc. The combined organic extracts were
dried over anh. Na.sub.2SO.sub.4, filtered and concentrated to
dryness in vacuo. The crude title compound (335 mg) was used in the
following step without further purification.
[0210] NMR (.sup.1H, CDCl.sub.3): .delta. 9.84 (s, 1H), 7.84 (d,
1H), 7.75 (d, 1H), 7.30 (d, 1H), 7.06 (d, 1H), 7.05 (s, 1H),
3.10-3.30 (m, 4 H), 2.55 (s, 3H).
[0211] MS (m/z): 333 [M+1].sup.+, 1Cl
[0212] Intermediate 17
3-[2-Chloro-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-3-yl]-propan--
1-ol
[0213] To a solution of intermediate 16 (335 mg, 1 mmol) in anh.
CH.sub.3OH (5 mL) were added CeCl.sub.3 (247 mg, 1 eq) and
NaBH.sub.4 (38 mg, 1 eq) at r.t., under N.sub.2. The reaction
mixture was stirred for 20 min. The solvent was removed in vacuo
and the residue was redissolved in EtOAc/H.sub.2O and the layers
were separated. The aqueous layer was extracted with EtOAc, and the
combined organic extracts were dried over anh. Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. Purification (silica gel,
cHex/EtOAc 8:2) of the crude afforded 277.4 mg of the title
compound as a clear oil.
[0214] NMR (.sup.1H, CDCl.sub.3): .delta. 7.90 (d, 1H), 7.76 (d,
1H), 7.36 (d, 1H), 7.12 (s, 1H), 7.07 (d, 1H), 3.70 (m, 2H), 2.90
(t, 2H), 2.58 (s, 3H), 2.20 (bt, 1H), 2.04 (m, 2H).
[0215] MS (m/z): 335 [M+1].sup.+, 1Cl
[0216] Intermediate 18
3-[3-(tert-Butyl-dimethyl-silanyloxy)-propyl]-2-chloro-6-methyl-4-(3-thiaz-
ol-2-yl-pyrazol-1-yl)-pyridine
[0217] To a solution of intermediate 17 (277.4 mg, 0.83 mmol) in
anh. DMF (7 mL) was added imidazole (621 mg, 11 eq),
tert-butyldimethylsilyl chloride (350 mg, 2.8 eq) and a catalytic
amount of DMAP at 0.degree. C. under N.sub.2. The reaction mixture
was stirred at room temperature for 2 hr. Then a saturated aqueous
solution of NH.sub.4Cl was added to the reaction mixture and it was
extracted with EtOAc. The combined organic extracts were dried over
anh. Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude product was purified by flash chromatography (silica gel,
cHex/EtOAc 7:3) to give 347 mg of the title compound as a yellow
oil.
[0218] NMR (.sup.1H, CDCl.sub.3): .delta. 7.89 (d, 1H), 7.81 (d,
1H), 7.34 (d, 1H), 7.20 (s, 1H), 7.08 (d, 1H), 3.66 (t, 2H), 2.86
(m, 2H), 2.57 (s, 3H), 1.89 (m, 2H), 0.86 (s, 9H), -0.006 (s,
6H).
[0219] MS (m/z): 449 [M].sup.+, 1Cl
[0220] Intermediate 19
(2,4-Bis-trifluoromethyl-phenyl)-[3-[3-(tert-butyl-dimethyl-silanyloxy)-pr-
opyl]-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-2-yl]-amine
[0221] To a vial containing Pd(dba).sub.3 (17 mg, 0.1 eq),
2-(Dicyclohexylphosphino)-2'-methylbiphenyl (20 mg, 0.3 eq) and
K.sub.3PO.sub.4 (103 mg, 2.7 eq), at r.t., under N.sub.2, were
added a solution of intermediate 18 (80 mg, 0.18 mmol) in anh. DME
(0.5 mL) and a solution of 2,4-bis(trifluoromethyl)aniline (82 mg,
2 eq) in dry DME (0.5 mL). The reaction mixture was submitted to
microwave irradiation five times (3.times.10 min+30 min+60 min)
with these observed parameters: P=110 W; T=100.degree. C., p=18
psi. The solution was diluted with water and extracted with EtOAc.
The combined organic extracts were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated to dryness in vacuo.
The crude product was purified by flash chromatography (silica gel,
cHex/Et.sub.2O 7:3) to give 49 mg of the title compound as a yellow
oil.
[0222] NMR (.sup.1H, CDCl.sub.3): .delta. 8.58 (d, 1H), 7.89 (d,
1H), 7.85 (d, 1H), 7.77 (dd, 1H), 7.76 (d, 1H), 7.34 (d, 1H), 7.23
(bs, 1H), 7.08 (d, 1H), 6.86 (s, 1H), 3.67 (t, 2H), 2.69 (m, 2H),
2.3 (s, 3H), 1.90 (m, 2H), 0.8 (s, 9H), -0.02 (s, 6H).
[0223] MS (m/z): 642 [M+1].sup.+
[0224] Intermediate 20
3-[2-(2,4-Bis-trifluoromethyl-phenylamino)-6-methyl-4-(3-thiazol-2-yl-pyra-
zol-1-yl)-pyridin-3-yl]-propan-1-ol
[0225] To a solution of intermediate 19 (60 mg, 0.094 mmol) in anh.
THF (2 mL) was added TEA-3HF (0.046 mL, 3 eq). The reaction mixture
was stirred at room temperature for 12 hr. Then a saturated
solution of NH.sub.4Cl was added to the reaction mixture and it was
extracted with EtOAc. The combined organic extracts were dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
product was purified by flash chromatography (silica gel,
cHex/EtOAc 1:1) to give 34.6 mg of the title compound as a white
solid.
[0226] NMR (.sup.1H, CDCl.sub.3): .delta. 8.62 (d, 1H), 7.90 (d,
1H), 7.85 (bs, 1H), 7.76 (dd, 1H), 7.7 (d, 1H), 7.37 (bs, 1H), 7.36
(d, 1H), 7.07 (d, 1H), 6.83 (s, 1H), 3.73 (t, 2H), 2.73 (t, 2H),
2.52 (s, 3H), 2.04(m, 2H).
[0227] MS (m/z): 528 [M+1].sup.+
[0228] Intermediate 21
2,4-Bis-trifluoromethyl-phenyl)-[3-(3-bromo-propyl)-6-methyl-4-(3-thiazol--
2-yl-pyrazol-1-yl)-pyridin-2-yl]-amine
[0229] To a solution of intermediate 20 (34.6 mg, 0.066 mmol) in
anh. CH.sub.2Cl.sub.2 (1 mL) was added CBr.sub.4 (44 mg, 2 eq) and
PPh.sub.3 (34 mg, 2 eq). The reaction mixture was stirred at room
temperature for 1 hr. Then a saturated aqueous solution of
NaHCO.sub.3 was added to the reaction mixture and it was extracted
with EtOAc. The combined organic extracts were dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
product was purified by flash chromatography (silica gel,
cHex/EtOAc 7:3) to give 25.7 mg of the title compound as a white
solid.
[0230] NMR (.sup.1H, CDCl.sub.3): .delta. 8.6 (d, 1H), 7.90 (d,
1H), 7.87 (d, 1H), 7.77 (m, 1H), 7.37 (d, 1H), 7.15 (bs, 1H), 7.10
(d, 1H), 6.84 (s, 1H), 3.47 (t, 2H), 2.78 (m, 2H), 2.52 (s, 3H),
2.3 (m, 2H).
[0231] MS (m/z): 590 [M].sup.+, 1Br; 510 [M-Br].sup.+
[0232] Intermediate 22
4-[3-[2-(tert-Butyl-dimethyl-silayloxy)-ethyl]-6-methyl-4-(3-thiazol-2-yl--
pyrazol-1-yl)-pyridin-2-ylamino]-3-methyl-benzonitrile
[0233] To a solution of intermediate 8 (186 mg, 0.43 mmol) in anh.
DME (1 mL) were added Pd.sub.2(dba).sub.3 (39 mg, 0.1 eq),
2-(dicyclohexylphosphino)-2'-methylbiphenyl (47 mg, 0.3 eq),
K.sub.3PO.sub.4 (246 mg, 2.6 eq) and 3-methyl-4-amino benzonitrile
(113 mg, 2 eq) and the reaction mixture was submitted to microwave
irradiation (150 W, 100.degree. C., 60 psi) for 20 min. The
reaction was then quenched with an aqueous solution of saturated
NH.sub.4Cl, extracted with EtOAc, washed with brine, dried over
anh. Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude product was purified by flash chromatography (silica gel,
cHex/EtOAc 8:2) to give 61 mg of the title compound as a white
solid.
[0234] NMR (.sup.1H, CDCl.sub.3): .delta. 8.30 (d, 1H), 8.06 (bs,
1H), 7.89 (d, 1H), 7.78 (d, 1H), 7.46 (dd, 1H), 7.44 (d, 1H), 7.36
(d, 1H), 7.09 (d, 1H), 6.81 (s, 1H), 4.34 (m, 2H), 2.82 (t, 2H),
2.56 (s, 3H), 2.36 (s, 3H), 0.85 (s, 9H), 0.02 (s, 6H).
[0235] MS (m/z): 531 [MH].sup.+.
[0236] Intermediate 23
4-[3-(2-Hydroxy-ethyl)-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-2--
ylamino]-3-methyl-benzonitrile
[0237] To a solution of intermediate 22 (61 mg, 0.115 mmol) in anh.
THF (2 mL) was added Et.sub.3N.3HF (0.056 ml, 3 eq) and the
reaction mixture was stirred for 15 hr at room temperature. The
reaction was then quenched with an aqueous solution of saturated
NH.sub.4Cl, extracted with EtOAc, washed with brine, dried over
anh. Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
product was purified by flash chromatography (silica gel,
cHex/EtOAc 1:1) to give 46 mg of the title compound as a white
solid.
[0238] NMR (.sup.1H, CDCl.sub.3): .delta. 8.39 (bs, 1H), 8.14 (d,
1H), 7.90 (d, 1H), 7.79 (d, 1H), 7.46 (m, 2H), 7.36 (d, 1H), 7.09
(d, 1H), 6.82 (s, 1H), 4.34 (m, 2H), 2.83 (t, 2H), 2.54 (s, 3H),
2.34 (s, 3H).
[0239] MS (m/z): 417 [MH].sup.+.
[0240] Intermediate 24
4-[3-(2-Hydroxy-ethyl)-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-pyridin-2--
ylamino]-3-trifluoromethyl-benzonitrile
[0241] To a solution of intermediate 8 (106 mg, 0.244 mmol) in anh.
DME (1 mL) were added Pd.sub.2(dba).sub.3 (22 mg, 0.1 eq),
2-(dicyclohexylphosphino)-2'-methylbiphenyl (27 mg, 0.3 eq),
K.sub.3PO.sub.4 (140 mg, 2.7 eq) and 3-trifluoromethyl-4-amino
benzonitrile (91 mg, 2 eq) and the reaction mixture was submitted
to microwave irradiation (150 W, 100.degree. C., 60 psi) for 20
min. The reaction was then quenched with an aqueous solution of
saturated NH.sub.4Cl, extracted with EtOAc, washed with brine,
dried over anh. Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The product was purified by flash chromatography (silica
gel, cHex[EtOAc 8:2) and the isolated product containing some
unreacted aniline was used in the next step without further
purification.
[0242] To a solution of the mixture obtained above (120 mg) in anh.
THF (5 mL) was added Et.sub.3N.3HF (0.063 ml, 3 eq) and the
reaction was stirred for 15 hr at room temperature. The reaction
was then quenched with an aqueous solution of saturated NH.sub.4Cl,
extracted with EtOAc, washed with brine, dried over anh.
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The product
was purified by flash chromatography (silica gel, cHex[EtOAc 1:1).
to give 40 mg of the title compound as a white solid.
[0243] NMR (.sup.1H, CDCl.sub.3): .delta. 8.81 (bs, 1H), 8.22 (d,
1H), 7.90 (d, 1H), 7.87 (d, 1H), 7.79 (d, 1H), 7.68 (dd, 1H), 7.37
(d, 1H), 7.17 (d, 1H), 6.92 (s, 1H), 4.26 (q, 2H), 2.87 (t, 2H),
2.54 (s, 3H), 2.63 (t, 1H).
[0244] MS (m/z): 471 [MH].sup.+.
[0245] Intermediate 25
[3-[2-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-6-methyl-4-(3-thiazol-2-yl-p-
yrazol-1-yl)-pyridin-2-yl]-(2-methyl-4-trifluoromethoxy-phenyl)-amine
[0246] To a solution of intermediate 8 (110 mg, 0.253 mmol) in anh.
DME (1 mL), at r.t., under N.sub.2, were added Pd.sub.2(dba).sub.3
(23 mg, 0.1 eq), 2-(dicyclohexylphosphino)-2'-methylbiphenyl (28
mg, 0.3 eq), K.sub.3PO.sub.4 (145 mg, 2.7 eq) and
2-methyl-4-trifluoromethyl aniline (97 mg, 2 eq) and the reaction
mixture was sumitted to microwave irradiation(150 W, 100.degree.
C., 60 psi) for 20 min. The reaction was then quenched with an
aqueous solution of saturated NH.sub.4Cl, extracted with EtOAc,
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The product was purified by flash
chromatography (silica gel, cHex/EtOAc 7:3) to give 80 mg of the
title compound as a yellow oil.
[0247] NMR (.sup.1H, CDCl.sub.3): .delta. 8.05 (d, 1H), 7.83 (bs,
1H), 7.78 (d, 1H), 7.7 (d, 1H), 7.46 (dd, 1H), 7.44 (d, 1H), 7.36
(d, 1H), 7.09 (d, 1H), 6.81 (s, 1H), 4.34 (m, 2H), 2.82 (t, 2H),
2.56 (s, 3H), 2.36 (s, 3H), 0.85 (s, 9H), 0.023 (s, 6H).
[0248] MS (m/z): 590 [MH].sup.+.
[0249] Intermediate 26
2-[6-Methyl-2-(2-methyl-4-trifluoromethoxy-phenylamino)-4-(3-thiazol-2-yl--
pyrazol-1-yl)-pyridin-3-yl]-ethanol
[0250] To a solution of intermediate 25 (80 mg, 0.135 mmol) in anh.
THF (2 mL) was added Et.sub.3N.3HF (66 .mu.L, 8 eq) and the
reaction mixture was stirred for 15 hr at room temperature. The
reaction was then quenched with an aqueous solution of saturated
NH.sub.4Cl, extracted with EtOAc, washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The product
was purified by flash chromatography (silica gel, cHex/EtOAc 1:1 )
to give 48 mg of the title compound as a colorless oil.
[0251] NMR (.sup.1H, CDCl.sub.3): .delta. 7.91 (bs, 1H), 7.85 (d,
1H), 7.7 (d, 1H), 7.65 (d, 1H), 7.30 (d, 1H), 7.15-6.95 (m, 3H),
6.65 (s, 1H), 4.34 (m, 2H), 2.83 (t, 2H), 2.54 (s, 3H), 2.34 (s,
3H).
Example 1
[0252] Synthesis of representative compounds of structure (IIIa)
##STR12##
Example 1-1
1-(2,4-Bis-trifluoromethyl-phenyl)-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl-
)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine
[0253] To a solution of intermediate 10 (40 mg, 0.078 mmol) in anh.
CH.sub.2Cl.sub.2 (2 mL), at r.t., under N.sub.2, were added
CBr.sub.4 (52 mg, 2 eq) and PPh.sub.3 (41 mg, 2 eq) and the
reaction mixture was stirred for 3 hr. The reaction was then
quenched with an aqueous solution of saturated NaHCO.sub.3,
extracted with EtOAc, washed with brine, dried over anh.
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The product
was purified by flash chromatography (silica gel, cHex/EtOAc 2:1)
to give 18 mg of the title compound as a white solid.
[0254] Alternatively:
Example 1-1
1-(2,4-Bis-trifluoromethyl-phenyl)-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl-
)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine
[0255] To a mixture of tris(dibenzylidenacetone)palladium(0) (3.2
mg, 0.1 eq), 2-(dicyclohexylphosphino)-2'-methylbiphenyl (3.8 mg,
0.3 eq) and K.sub.3PO.sub.4 (20 mg, 2.8 eq) in a crimp cap
microwave vial was added a solution of intermediate 14 (10 mg,
0.035 mmol) and 2,4-bis(trifluoromethyl)-bromobenzene (6 .mu.L, 1
eq) in anh. DME (1 mL), under N.sub.2. The reaction mixture was
submitted to microwave irradiation for two cycles (2.times.10 min)
with these observed parameters: P=150 W; T=100.degree. C., p=60 psi
Then water (1 mL) was added and the product was extracted with
EtOAc. The combined organic extracts were washed with sat. aq. NaCl
(5 mL) and dried over anh. Na.sub.2SO.sub.4. The solids were
filtered and the solvent evaporated to give a crude product, which
was purified by flash chromatography (silica gel, cHex/EtOAc 7:3).
The title compound was obtained as a colourless oil (1 mg, 0.002
mmol).
Example 1-2
3-Methyl-4-[6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-2,3-dihydro-1H-pyrrol-
o[2,3-b]pyridin-1-yl]-benzonitrile
[0256] To a solution of intermediate 23 (44 mg, 0.106 mmol) in anh.
CH.sub.2Cl.sub.2 (2 mL) were added CBr.sub.4 (71 mg, 2 eq) and
PPh.sub.3 (60 mg, 2 eq) and the reaction mixture was stirred for 3
hr at room temperature. The reaction was then quenched with an
aqueous solution of saturated NaHCO.sub.3, extracted with EtOAc,
washed with brine, dried over anh. Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The product was purified by flash
chromatography (silica gel, cHex/EtOAc 4:1) to give 18 mg of the
title compound as a white solid.
Example 1-3
4-[6-Methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]p-
yridin-1-yl]-3-trifluoromethyl-benzonitrile
[0257] To a solution of intermediate 24 (40 mg, 0.085 mmol) in anh.
CH.sub.2Cl.sub.2 (2 mL) were added CBr.sub.4 (56 mg, 2 eq) and
PPh.sub.3 (45 mg, 2 eq) and the reaction mixture was stirred for 3
hr at room temperature. The reaction was then quenched with an
aqueous solution of saturated NaHCO.sub.3, extracted with EtOAc,
washed with brine, dried over anh. Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The product was purified by flash
chromatography (silica gel, cHex/EtOAc 2:1) to give 13 mg of the
title compound as a white solid.
Example 1-4
6-Methyl-1-(2-methyl-4-trifluoromethoxy-phenyl)-4-(3-thiazol-2-yl-pyrazol--
1-yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine
[0258] To a solution of intermediate 26 (48 mg, 0.101 mmol) in anh.
CH.sub.2Cl.sub.2 (2 mL) were added CBr.sub.4 (66 mg, 2 eq) and
PPh.sub.3 (53 mg, 2 eq) and the reaction mixture was stirred for 3
hr at room temperature. The reaction was then quenched with an
aqueous solution of saturated NaHCO.sub.3, extracted with EtOAc,
washed with brine, dried over anh. Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The product was purified by flash
chromatography (silica gel, cHex/EtOAc 8:2) to give 10 mg of the
title compound as a white solid.
Example 1-5
1-(4-Methoxy-2-methyl-phenyl)-6-methyl-4-(3-thiazol-2-yl-pyrazol-1-yl)-2,3-
-dihydro-1H-pyrrolo[2,3-b]pyridine
[0259] To a solution of intermediate 28 (31 mg, 0.075 mmol, 1 eq.)
in DCM dry (5 ml), was added CBr.sub.4 (53 mg, 0.16 mmol, 2.1 eq.)
and triphenylphosphine (42 mg, 0.16 mmol, 2.1 eq.) under N.sub.2.
The reaction mixture was stirred at RT for 15 hrs. Then water (10
ml) was added and the aqueous phase was extracted with EtOAc (20
ml). The organic layer was dried over Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude was purified by flash
chromatography (Eluents: cyclohexane/ethyl acetate 7:3) to give 5.2
mg of VSAF/6274/4/1 as a colorless oil.
Example 1-6
1-(2,4-Bis-trifluoromethyl-phenyl)-6-methyl-4-(3-morpholin-4-yl-pyrazol-1--
yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine
[0260] Prepared analogously to example 1-1 using
4-(1H-pyrazol-3-yl)-morpholine (J. Org. Chem., 1984, 269-276)
instead of 2-(1H-pyrazol-3-yl)-thiazole in the preparation of
intermediate 2.
Example 1-7
1-(2,4-Bis-trifluoromethyl-phenyl)-6-methyl-4-(3-pyridin-2-yl-pyrazol-1-1--
yl)-2,3-dihydro-1H-pyrrolo[2,3-b]pyridine
[0261] Prepared analogously to example 1-1 using
2-(1H-Pyrazol-3-yl)-pyridine (commercially available) instead of
2-(1H-pyrazol-3-yl)-thiazole in the preparation of intermediate
2.
Example 1-8
4-[1,3']Bipyrazolyl-1'-yl-1-(2,4-bis-trifluoromethyl-phenyl)-6-methyl-2,3--
dihydro-1H-pyrrolo[2,3-b]pyridine
[0262] Prepared analogously to example 1-1 using
1'H-[1,3']bipyrazolyl (from 1H-pyrazol-3-ylamine: J. Heterocycl.
Chem., 1983, 1629-1639; then J. Heterocycl. Chem., 1989, 733-738)
instead of 2-(1H-pyrazol-3-yl)-thiazole in the preparation of
intermediate 2.
[0263] All the analytical data are set forth in the following Table
1-1. ##STR13## TABLE-US-00001 TABLE 1-1 Cpd. No. R R.sub.1
R.sub.2--R.sub.3-- Analytical Data 1-1 2,4-bistrifluoro-
methylphenyl CH.sub.3 ##STR14## NMR (.sup.1H, DMSO): .delta. 8.64
(d, 1H), 8.17 (dd, 1H), 8.13 (d, 1H), 7.94 (d, 1H), 7.84 (d, 1H),
7.80 (d, 1H), 7.09 (m, 2H), 3.97 (t, 2H), 3.56 (t, 2H), 2.24 (s,
3H). MS (m/z): 496 [MH].sup.+. 1-2 2-methyl-4-cyano CH.sub.3
##STR15## NMR (.sup.1H, DMSO): .delta. 8.02 (d, 1H), 7.90 (d, 1H),
7.59 (d, 1H), 7.53 (dd, 1H), 7.41 (d, 1H), 7.38 (d, 1H), 6.76 (s,
1H), 4.04 (t, 2H), 3.62 (t, 2H), 2.41 (s, 3H), 2.33 (s, 3H). MS
(m/z): 399 [MH].sup.+. 1-3 2-trifluoromethyl- 4-cyano CH.sub.3
##STR16## NMR (.sup.1H, CDCl.sub.3): .delta. 8.00 (m, 2H), 7.9-7.8
(d+d, 2H), 7.65 (d, 1H), 7.35 (d, 1H), 7.10 (d, 1H), 6.80 (s, 1H),
4.00 (t, 2H), 3.60 (t, 2H), 3.56 (t, 2H), 2.40 (s, 3H). MS (m/z):
453 [MH].sup.+. 1-4 2-methyl-4-tri- fluoromethoxy CH.sub.3
##STR17## NMR (.sup.1H, DMSO): .delta. 8.01 (d, 1H), 7.89 (d, 1H),
7.5 (d, 1H), 7.45 (dd, 1H), 7.20 (d, 1H), 7.1 (dd, 2H), 6.65 (s,
1H), 3.9 (t, 2H), 3.56 (t, 2H), 2.32 (s, 3H), 2.25 (s, 3H). MS
(m/z): 458.5 [MH].sup.+. 1-5 2-methyl-4-methoxy CH.sub.3 ##STR18##
NMR (.sup.1H, CDCl.sub.3): .delta. 7.97 (d, 1H), 7.86 (d, 1H), 7.33
(d, 1H), 7.17 (d, 1H), 7.10 (d, 1H), 6.78 (m, 2H), 6.61 (s, 1H),
3.90 (t, 2H), 3.80 (s, 3H), 3.52 (t, 2H), 2.34 (s, 3H), 2.23 (s,
3H). MS (m/z): 404 [MH].sup.+
Example 2
[0264] Synthesis of representative compounds of structure (IIIb)
##STR19##
[0265] All the analytical data are set forth in the following Table
2-1.
Example 2-1
1-(2,4-Bis-trifluoromethyl-phenyl)-7-methyl-5-(3-thiazol-2-yl-pyrazol-1-yl-
)-1,2,3,4-tetrahydro-[1,8]naphthyridine
[0266] To a solution of intermediate 21 (24.8 mg, 0.042 mmol) in
anh. N-metylpyrrolidinone (2 mL) was added Et.sub.3N (12 .mu.L, 2
eq) under N.sub.2. The reaction mixture was submitted to microwave
irradiation for 10 min with these observed parameters: P=90 W;
T=99.degree. C., p=6 psi. Then a saturated aqueous solution of
NH.sub.4Cl was added to the reaction mixture and the aqueous phase
was extracted with EtOAc. The combined organic extracts were washed
with a saturated solution of NH.sub.4Cl (3.times.), dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
product was purified by SCX Column (Eluents: CH.sub.2Cl.sub.2, MeOH
and a solution of conc. NH.sub.4OH in MeOH (25%) to elute the
desired product) to give 18.4 mg of the title compound as a white
foam. TABLE-US-00002 TABLE 2-1 Cpd. No. R R.sub.1
R.sub.2--R.sub.3-- Analytical Data 2-1 2,4-bistrifluoro-
methylphenyl CH.sub.3 ##STR20## NMR (.sup.1H, CDCl.sub.3): .delta.
8.02 (bs, 1H), 7.89 (d, 1H), 7.88 (dd, 1H), 7.74 (d, 1H), 7.46 (d,
1H), 7.35 (d, 1H), 7.06 (d, 1H), 6.59 (s, 1H), 3.66 (bm, 2H), 2.97
(bm, 1H), 2.85 (bm, 1H), 2.18 (s, 3H), 2.09 (bm, 1H), 2.04 (bm,
1H). MS (m/z): 510 [M+ 1].sup.+
Example 3
[0267] CRF Binding Activity
[0268] CRF binding affinity has been determined in vitro by the
compounds' ability to displace .sup.125I-oCRF and
.sup.125I-Sauvagine for CRF1 and CRF2 SPA, respectively, from
recombinant human CRF receptors expressed in Chinese Hamster Ovary
(CHO) cell membranes. For membrane preparation, CHO cells from
confluent T-flasks were collected in SPA buffer (HEPES/KOH 50 mM,
EDTA 2 mM; MgCl.sub.2 10 mM, pH 7.4.) in 50 mL centrifuge tubes,
homogenized with a Polytron and centrifuged (50,000 g for 5 min at
4.degree. C.: Beckman centrifuge with JA20 rotor). The pellet was
resuspended, homogenized and centrifuged as before.
[0269] The SPA experiment has been carried out in Optiplate by the
addition of 100 .mu.L the reagent mixture to 1 .mu.L of compound
dilution (100% DMSO solution) per well. The assay mixture was
prepared by mixing SPA buffer, WGA SPA beads (2.5 mg/mL), BSA (1
mg/mL) and membranes (50 and 5 .mu.g of protein/mL for CRF1 and
CRF2 respectively) and 50 pM of radioligand.
[0270] The plate was incubated overnight (>18 hrs) at room
temperature and read with the Packard Topcount with a WGA-SPA
.sup.125I counting protocol.
Example 4
[0271] CRF Functional Assay
[0272] Compounds of the invention were characterised in a
functional assay for the determination of their inhibitory effect.
Human CRF-CHO cells were stimulated with CRF and the receptor
activation was evaluated by measuring the accumulation of cAMP.
[0273] CHO cells from a confluent T-flask were resuspended with
culture medium without G418 and dispensed in a 96-well plate,
25,000 c/well, 100 .mu.L/well and incubated overnight. After the
incubation the medium was replaced with 100 .mu.L of cAMP IBMX
buffer warmed at 37.degree. C. (5 mM KCl, 5 mM NaHCO.sub.3, 154 mM
NaCl, 5 mM HEPES, 2.3 mM CaCl.sub.2, 1 mM MgCl.sub.2; 1 g/L
glucose, pH 7.4 additioned by 1 mg/mL BSA and 1 mM IBMX) and 1
.mu.L of antagonist dilution in neat DMSO. After 10 additional
minutes of incubation at 37.degree. C. in a plate incubator without
CO2, 1 .mu.L of agonist dilution in neat DMSO was added. As before,
the plate was incubated for 10 minutes and then cAMP cellular
content was measured by using the Amersham RPA 538 kit.
[0274] All publications, including but not limited to patents and
patent applications, cited in this specification are herein
incorporated by reference as if each individual publication were
specifically and individually indicated to be incorporated by
reference herein as though fully set forth.
[0275] It is to be understood that the present invention covers all
combinations of particular and preferred groups described herein
above.
[0276] The application of which this description and claims forms
part may be used as a basis for priority in respect of any
subsequent application. The claims of such subsequent application
may be directed to any feature or combination of features described
herein. They may take the form of product, composition, process, or
use claims and may include, by way of example and without
limitation, the following claims:
* * * * *