U.S. patent application number 11/246663 was filed with the patent office on 2006-05-25 for therapeutic benzothiazole compounds.
Invention is credited to Bernard Barlaam, Peter Bernstein, Cathy Dantzman, Paul Warwick.
Application Number | 20060111408 11/246663 |
Document ID | / |
Family ID | 26655351 |
Filed Date | 2006-05-25 |
United States Patent
Application |
20060111408 |
Kind Code |
A1 |
Barlaam; Bernard ; et
al. |
May 25, 2006 |
Therapeutic benzothiazole compounds
Abstract
Compounds of the formula (I) ##STR1## for use as an estrogen
receptor -.beta.-selective ligand are described wherein: X is O or
S; and R.sup.1, R.sup.3-R.sup.6 are as described in the
specification. The use of these compounds in treating Alzheimer's
disease, anxiety disorders, depressive disorders, osteoporosis,
cardiovascular disease, rheumatoid arthritis and prostate cancer is
described; as are processes for making them.
Inventors: |
Barlaam; Bernard; (Reims,
FR) ; Bernstein; Peter; (Wilmington, DE) ;
Dantzman; Cathy; (Wilmington, DE) ; Warwick;
Paul; (Wilmington, DE) |
Correspondence
Address: |
WHITE & CASE LLP;PATENT DEPARTMENT
1155 AVENUE OF THE AMERICAS
NEW YORK
NY
10036
US
|
Family ID: |
26655351 |
Appl. No.: |
11/246663 |
Filed: |
January 3, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10450927 |
Nov 26, 2003 |
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PCT/SE01/02855 |
Dec 19, 2001 |
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11246663 |
Jan 3, 2006 |
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Current U.S.
Class: |
514/367 ;
548/152 |
Current CPC
Class: |
A61P 9/02 20180101; A61P
19/10 20180101; C07D 413/04 20130101; A61P 25/28 20180101; A61P
29/00 20180101; A61P 25/24 20180101; C07D 277/66 20130101; C07D
417/04 20130101; A61P 25/22 20180101; A61P 19/02 20180101; A61P
25/00 20180101; A61P 5/30 20180101; C07D 263/57 20130101; A61P
35/00 20180101 |
Class at
Publication: |
514/367 ;
548/152 |
International
Class: |
A61K 31/428 20060101
A61K031/428; C07D 417/02 20060101 C07D417/02; C07D 277/60 20060101
C07D277/60 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 22, 2000 |
SE |
0004825-6 |
Dec 22, 2000 |
SE |
0004826-4 |
Claims
1-17. (canceled)
18. A compound of the formula (I) ##STR109## or a pharmaceutically
acceptable salt thereof, wherein: X is S; R.sup.1 is
C.sub.1-8alkyl, phenyl or a 5- or 6-membered ring heterocycle
containing 1, 2 or 3 heteroatoms each independently selected from
O, N and S and additionally having 0 or 1 oxo groups and 0 or 1
fused benzo rings, wherein the C.sub.1-8alkyl or heterocycle is
substituted by 0, 1, 2 or 3 substituents selected from --R.sup.a,
--OR.sup.a, --SR.sup.a, --NR.sup.aR.sup.a, --CO.sub.2R.sup.a,
--OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
and C.sub.1-3haloalkyl and the phenyl is substituted by 0, 1, 2 or
3 substituents selected from --R.sup.a, --CO.sub.2R.sup.a,
--OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O) R.sup.a, --NR.sup.aS(.dbd.O) R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
and C.sub.1-3haloalkyl; R.sup.3 is --R.sup.a, --OR.sup.a,
--SR.sup.a, --NR.sup.aR.sup.a, --CO.sub.2R.sup.a,
--OC(.dbd.O)R.sup.a, C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
or C.sub.1-3haloalkyl; or R.sup.3 is C.sub.1-3alkyl containing 1 or
2 substituents selected from --OR.sup.a, --SR.sup.a,
--NR.sup.aR.sup.a, --CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, --NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
--C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a,
halogen, cyano and nitro; R.sup.4 is --R.sup.a, --OR.sup.a,
--SR.sup.a, --NR.sup.aR.sup.a, --CO2R.sup.a, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, --NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
--C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a,
halogen, cyano, nitro or C.sub.1-3haloalkyl; R.sup.5 is --R.sup.a,
--OR.sup.a, --SR.sup.a, --NR.sup.aR.sup.a, --CO.sub.2R.sup.a,
--OC(.dbd.O)R.sup.a, C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
or C.sub.1-3haloalkyl; R.sup.6 is --R.sup.a, --OR.sup.a,
--SR.sup.a, --NR.sup.aR.sup.a, --CO.sub.2R.sup.a,
--OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
and or C.sub.1-3haloalkyl; or R.sup.6 is C.sub.1-3alkyl containing
1 or 2 substituents selected from --OR.sup.a, --SR.sup.a,
--NR.sup.aR.sup.a, --CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, --NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
--C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a,
halogen, cyano and nitro; and R.sup.a is H, C.sub.1-6alkyl,
C.sub.1-3haloalkyl, phenyl or benzyl; with the provisos that when
the compound is in free base form and: a) R.sup.1 is phenyl, the
benzene ring of the benzthiazole is not substituted by 4-methoxy,
5,6-dimethoxy, 6-hydroxy or 6-methoxy; and b) R.sup.1 is
4-methylphenyl, the benzene ring of the benzthiazole is not
unsubstituted and is not substituted by 4-, 5- or 6-fluoro, 4-, 6-
or 7-methoxy, 5-chloro, 4-, 5-, 6- or 7-hydroxy, 4-, 5-, 6- or
7-acetoxy or 6-nitro; and R.sup.1 is not 4-chloro- or
4-fluorophenyl when the benzene ring of the benzthiazole is
substituted by 5-hydroxy or 5-mercapto.
19. (canceled)
20. (canceled)
21. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.1 is
unsubstituted or substituted phenyl.
22. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.1 is an
unsubstituted or substituted 5- or 6-membered ring heterocycle.
23. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.1 is
halophenyl, C.sub.1-4alkylphenyl, cyanophenyl or
trifluoromethylphenyl.
24. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.3 is
halo, cyano, carbamoyl or C.sub.1-6alkyl.
25. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.3 is
hydrogen.
26. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.4 is
halo, hydroxy or C.sub.1-6alkoxy.
27. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.4 is
hydrogen.
28. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.5 is
halo, hydroxy or C.sub.1-6alkoxy.
29. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.5 is
hydrogen.
30. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.6 is
halo, C.sub.1-4alkyl, trifluoromethyl, hydroxy, C.sub.1-4alkoxy,
carboxy, C.sub.1-4alkoxycarbonyl, cyano, halomethyl,
cyanoC.sub.1-4alkyl, carbamoyl, methylcarbamoyl or
dimethylcarbamoyl.
31. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.6 is
hydrogen.
32. (canceled)
33. The compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 wherein R.sup.6 is
cyano or carboxy; R.sup.4 is hydroxy; and R.sup.3 and R.sup.5 are
both hydrogen.
34. A pharmaceutical composition comprising a compound of the
formula (I) or a pharmaceutically acceptable salt thereof as
defined in claim 18 and a pharmaceutically acceptable carrier.
35. (canceled)
36. A method for achieving selective targeting of the
.beta.-estrogen receptor, which comprises administering an
effective amount of a compound of the formula (I) or a
pharmaceutically acceptable salt thereof according to claim 18.
37. A method for treating Alzheimer's disease, anxiety disorders,
depressive disorders, osteoporosis, cardiovascular disease,
rheumatoid arthritis or prostate cancer, which comprises
administering an effective amount of a compound of the formula (I)
or a pharmaceutically acceptable salt thereof according to claim 18
to a patient in need of such treatment.
38. (canceled)
39. A process for preparing a compound of the formula (I) or a
pharmaceutically acceptable salt thereof, as defined in claim 18
which comprises: a) cyclizing a compound of the formula: ##STR110##
wherein X, R.sup.1, and R.sup.3-R.sup.6 are as defined in claim 18
and L is hydrogen or a leaving group; or b) cyclizing a compound of
the formula: ##STR111## wherein R.sup.1, R.sup.3-R.sup.6 and X are
as defined in claim 18 and L is hydrogen or a leaving group; or c)
cyclizing a compound of the formula: ##STR112## wherein R.sup.1 and
R.sup.3-R.sup.6 are as defined in claim 18; and thereafter,
desired, forming a pharmaceutically acceptable salt.
40. A method for treating a condition for which ERT has a
beneficial effect, which comprises administering an effective
amount of a compound of the formula (I) or a pharmaceutically
acceptable salt thereof according to claim 18 to a patient in need
of such treatment.
Description
[0001] This application is a divisional of U.S. application Ser.
No. 10/450,927, which is the National Stage of International
Application No. PCT/SE01/02855, filed Dec. 19, 2001.
TECHNICAL FIELD
[0002] The present invention is directed to a series of ligands,
and more particularly to estrogen receptor-.beta. ligands which
have better selectivity than estrogen for the estrogen
receptor-.beta. over the estrogen receptor-.alpha., as well as to
methods for their production and use in the treatment of diseases
related to the estrogen receptor-.beta., specifically, Alzheimer's
disease, anxiety disorders, depressive disorders, osteoporosis,
cardiovascular disease, rheumatoid arthritis, or prostate
cancer.
BACKGROUND
[0003] Estrogen-replacement therapy ("ERT") reduces the incidence
of Alzheimer's disease and improves cognitive function in
Alzheimer's disease patients (Nikolov et al. Drugs of Today,
34(11), 927-933 (1998)). ERT also exhibits beneficial effects in
osteoporosis and cardiovascular disease, and may have anxiolytic
and anti-depressant therapeutic properties. However, ERT shows
detrimental uterine and breast side effects that limit its use.
[0004] The beneficial effects of ERT in post-menopausal women is
echoed by beneficial effects of estrogen in models relevant to
cognitive function, anxiety, depression, bone loss, and
cardiovascular damage in ovariectomized rats. Estrogen also
produces uterine and breast hypertrophy in animal models
reminiscent of its mitogenic effects on these tissues in humans.
Specifically, experimental studies have demonstrated that estrogen
effects the central nervous system ("CNS") by increasing
cholinergic function, increasing neurotrophin/neurotrophin receptor
expression, altering amyloid precursor protein processing,
providing neuroprotection against a variety of insults, and
increasing glutamatergic synaptic transmission, among other
effects. The overall CNS profile of estrogen effects in
pre-clinical studies is consistent with its clinical utility in
improving cognitive function and delaying Alzheimer's disease
progression. Estrogen also produces mitogenic effects in uterine
and breast tissue indicative of its detrimental side effects on
these tissues in humans.
[0005] The estrogen receptor ("ER") in humans, rats, and mice
exists as two subtypes, ER-.alpha. and ER-.beta., which share about
a 50% identity in the ligand-binding domain (Kuiper et al.
Endocrinology 139(10) 4252-4263 (1998)). The difference in the
identity of the subtypes accounts for the fact that some small
compounds have been shown to bind preferentially to one subtype
over the other (Kuiper et al.).
[0006] In rats, ER-.beta. is strongly expressed in brain, bone and
vascular epithelium, but weakly expressed in uterus and breast,
relative to ER-.alpha.. Furthermore, ER-.alpha. knockout
(ERKO-.alpha.) mice are sterile and exhibit little or no evidence
of hormone responsiveness of reproductive tissues. In contrast,
ER-.beta. knockout (ERKO-.beta.) mice are fertile, and exhibit
normal development and function of breast and uterine tissue. These
observations suggest that selectively targeting ER-.beta. over
ER-.alpha. could confer beneficial effects in several important
human diseases, such as Alzheimer's disease, anxiety disorders,
depressive disorders, osteoporosis, and cardiovascular disease
without the liability of reproductive system side effects.
Selective effects on ER-.beta.-expressing tissues (CNS, bone, etc.)
over uterus and breast could be achieved by agents that selectively
interact with ER-.beta. over ER-.alpha..
[0007] It is a purpose of this invention to identify
ER-.beta.-selective ligands that are useful in treating diseases in
which ERT has therapeutic benefits.
[0008] It is another purpose of this invention to identify
ER-.beta.-selective ligands that mimic the beneficial effects of
ERT on brain, bone and cardiovascular function.
[0009] It is another purpose of this invention to identify
ER-.beta.-selective ligands that increase cognitive function and
delay Alzheimer's disease progression.
SUMMARY OF THE INVENTION
[0010] This present invention is directed to compounds having the
generic structure: ##STR2## These compounds are ER-.beta.-selective
ligands, which mimic ERT, but lack undesirable side effects of ERT
and are useful in the treatment or prophylaxis of Alzheimer's
disease, anxiety disorders, depressive disorders, osteoporosis,
cardiovascular disease, rheumatoid arthritis or prostate
cancer.
BRIEF DESCRIPTION OF THE DRAWINGS
[0011] FIG. 1 is a graph of absorbance values obtained in assays
for the binding to the estrogen receptor of the ER agonist
17-.beta.-estradiol (E) and the ER antagonist ICI182,780 (A) for
cells transfected either with .alpha.ER or .beta.ER.
[0012] FIG. 2 shows typical concentration-response curves,
providing EC.sub.50 values, for binding to .alpha.ER and
.beta.ER.
DETAILED DESCRIPTION OF THE INVENTION
[0013] The present invention provides compounds of the formula (I)
##STR3##
[0014] for use as ER-.beta.-selective ligands:
wherein:
[0015] X is O or S;
[0016] R.sup.1 is C.sub.1-8alkyl, phenyl, benzyl or a 5- or
6-membered ring heterocycle containing 1, 2 or 3 heteroatoms each
independently selected from O, N and S and additionally having 0 or
1 oxo groups and 0 or 1 fused benzo rings, wherein the
C.sub.1-8alkyl, phenyl, benzyl or heterocycle is substituted by 0,
1, 2 or 3 substituents selected from -R.sup.a, --OR.sup.a,
SR.sup.a, NR.sup.aR.sup.a, CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, --NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a,
halogen, cyano, nitro and C.sub.1-3haloalkyl;
[0017] R.sup.3 is --R.sup.a, --OR.sup.a, --SR.sup.a,
--NR.sup.aR.sup.a, --CO.sub.2R, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
--C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a,
halogen, cyano, nitro and C.sub.1-3haloalkyl; or R.sup.3 is
C.sub.1-3alkyl containing 1 or 2 substituents selected from
--OR.sup.a, --SR.sup.a, --NR.sup.aR.sup.a, --CO.sub.2R.sup.a,
--OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano and
nitro;
[0018] R.sup.4 is --R.sup.a, --OR.sup.a, --SR.sup.a,
--NR.sup.aR.sup.a, --CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, --NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
--C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a,
halogen, cyano, nitro or C.sub.1-3haloalkyl;
[0019] R.sup.5 is --R, --OR.sup.a, --SR.sup.a, --NR.sup.aR.sup.a,
--CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
or C.sub.1-3haloalkyl;
[0020] R.sup.6 is --R.sup.a, --OR.sup.a, --SR.sup.a,
--NR.sup.aR.sup.a, --CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, --NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
--C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a,
halogen, cyano, nitro and C.sub.1-3haloalkyl; or R.sup.6 is
C.sub.1-3alkyl containing 1 or 2 substituents selected from
--OR.sup.a, --SR.sup.a, --NR.sup.aR.sup.a, --CO.sub.2R.sup.a,
--OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano and
nitro; and
[0021] R.sup.a is H, C.sub.1-6alkyl, C.sub.1-3haloalkyl, phenyl or
benzyl;
and pharmaceutically acceptable salts thereof.
[0022] In the above definitions, where R.sup.a appears twice in a
group, each may be separately selected from the possible
values.
[0023] These compounds are useful in treating disease conditions
related to the .beta.-estrogen receptor, more particularly in
treating Alzheimer's disease, anxiety disorders, depressive
disorders, osteoporosis, cardiovascular disease, rheumatoid
arthritis and prostate cancer.
[0024] In another aspect the present invention provides the use of
a compound of the formula (I) or a pharmaceutically acceptable salt
thereof for the manufacture of a medicament for the treatment
(including prophylaxis) of disease conditions related to the
.beta.-estrogen receptor, more particularly in treating Alzheimer's
disease, anxiety disorders, depressive disorders, osteoporosis,
cardiovascular disease, rheumatoid arthritis and prostate
cancer.
[0025] In a further aspect the present invention provides a method
of treating disease conditions related to the .beta.-estrogen
receptor, more particularly in treating Alzheimer's disease,
anxiety disorders, depressive disorders, osteoporosis,
cardiovascular disease, rheumatoid arthritis and prostate
cancer.
[0026] In one embodiment R.sup.1 is C.sub.1-8alkyl, phenyl, benzyl
or a 5- or 6-membered ring heterocycle containing 1, 2 or 3
heteroatoms each independently selected from O, N and S and
additionally having 0 or 1 oxo groups and 0 or 1 fused benzo rings,
wherein the C.sub.1-8alkyl, phenyl, benzyl or heterocycle is
substituted by 0, 1, 2 or 3 substituents selected from --R.sup.a,
--OR.sup.a, --SR.sup.a, --NR.sup.aR.sup.a, --CO.sub.2R.sup.a,
--OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
and C.sub.1-3haloalkyl.
[0027] In another embodiment R.sup.3 is C.sub.1-6alkyl, --OR.sup.a,
--SR.sup.a, --NR.sup.aR.sup.a, --CO.sub.2R.sup.a,
--OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
and C.sub.1-3haloalkyl; or R.sup.3 is C.sub.1-3alkyl containing 1
or 2 substituents selected from --OR.sup.a, --SR.sup.a,
--NR.sup.aR.sup.a, --CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, --NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
--C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a,
halogen, cyano and nitro.
[0028] In another embodiment R.sup.4 is --R.sup.a, --SR.sup.a,
--NR.sup.aR.sup.a, --CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, --NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
--C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, S(.dbd.O).sub.2R.sup.a,
halogen, cyano, nitro or C.sub.1-3haloalkyl.
[0029] In another embodiment R.sup.5 is --SR.sup.a,
--NR.sup.aR.sup.a, --CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, --NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
--C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, S(.dbd.O).sub.2R.sup.a,
halogen, cyano, nitro or C.sub.1-3haloalkyl.
[0030] In another embodiment R.sup.6 is C.sub.1-6alkyl, --OR.sup.a,
--SR.sup.a, --NR.sup.aR.sup.a, --CO.sub.2R.sup.a,
--OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
and C.sub.1-3haloalkyl; or R.sup.6 is C.sub.1-3alkyl containing 1
or 2 substituents selected from --OR.sup.a, --SR.sup.a,
--NR.sup.aR.sup.a, --CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a,
--C(.dbd.O)NR.sup.aR.sup.a, --NR.sup.aC(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O).sub.2R.sup.a,
--C(.dbd.O)R.sup.a, --S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a,
halogen, cyano and nitro.
[0031] In another embodiment R.sup.1 is phenyl or benzyl, wherein
the phenyl or benzyl is substituted by 0, 1, 2 or 3 substituents
selected from --R.sup.a, --OR.sup.a, --SR.sup.a, --NR.sup.aR.sup.a,
--CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
and C.sub.1-3haloalkyl. In a more specific embodiment, R.sup.1 is
4-hydroxyphenyl substituted by 0, 1 or 2 substituents selected from
--R.sup.a, --OR.sup.a, --SR.sup.a, --NR.sup.aR.sup.a,
--CO.sub.2R.sup.a, --OC(.dbd.O)R.sup.a, --C(.dbd.O)NR.sup.aR.sup.a,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aS(.dbd.O)R.sup.a,
--NR.sup.aS(.dbd.O).sub.2R.sup.a, --C(.dbd.O)R.sup.a,
--S(.dbd.O)R.sup.a, --S(.dbd.O).sub.2R.sup.a, halogen, cyano, nitro
and C.sub.1-3haloalkyl.
[0032] In one aspect X is S. In another aspect X is O.
[0033] R.sup.1 may be a 5- or 6-membered ring heterocycle,
unsubstituted or substituted as defined hereinabove; for example
the 5- or 6-membered ring may be thiophene, furan, pyrrolidinone,
pyridine, indazole or thiazolidinone. In a preferred aspect R.sup.1
is phenyl unsubstituted or substituted as defined hereinabove.
Examples of R.sup.1 being substituted phenyl include hydroxyphenyl
(for example 4-hydroxyphenyl or 3-hydroxyphenyl),
C.sub.1-4alkoxyphenyl (for example 4-methoxyphenyl or
3-methoxyphenyl), halophenyl (for example bromophenyl such as
2-bromophenyl or chlorophenyl such as 2-chlorophenyl),
C.sub.1-4alkylphenyl (for example methylphenyl such as
2-methylphenyl or 3-methylphenyl or ethylphenyl such as
2-ethylphenyl or propylphenyl such as 2-isopropylphenyl),
cyanophenyl (for example 2-cyanophenyl) or trifluoromethylphenyl
(for example 4-trifluoromethylphenyl).
[0034] In particular R.sup.1 is hydroxyphenyl.
[0035] In a particular aspect R.sup.3is halo, cyano, carbamoyl or
C.sub.1-6alkyl; more particularly halo for example chloro or bromo,
cyano, or C.sub.1-6 alkyl for example methyl or ethyl. In another
particular aspect R.sup.3 is hydrogen.
[0036] In a particular aspect R.sup.4 is halo, for example chloro
or bromo, hydroxy or C.sub.1-6 alkoxy, for example methoxy or
ethoxy; more particularly R.sup.4 is hydroxy or methoxy, for
example hydroxy. In another particular aspect R.sup.4 is
hydrogen.
[0037] In a particular aspect R.sup.5 is halo, for example chloro
or bromo, hydroxy or C.sub.1-6 alkoxy, for example methoxy or
ethoxy; more particularly R.sup.5 is hydroxy or methoxy, for
example hydroxy. In another particular aspect R.sup.5 is
hydrogen.
[0038] In a particular aspect R.sup.6 is halo, for example chloro
or bromo, C.sub.1-4alkyl for example, methyl or ethyl,
trifluoromethyl, hydroxy, C.sub.1-4alkoxy, for example methoxy or
ethoxy, carboxy, C.sub.1-4alkoxycarbonyl, for example
methoxycarbonyl, cyano, halomethyl, for example bromomethyl,
cyanoC.sub.1-4alkyl, for example cyanomethyl, carbamoyl,
methylcarbamoyl or dimethylcarbamoyl. In another particular aspect
R.sup.6 is hydrogen. In one embodiment R.sup.6 is halo, cyano or
C.sub.1-6alkyl.
[0039] Preferred benzoxazoles are those wherein R.sup.1 is
4-hydroxyphenyl or 3-chloro-4-hydroxy phenyl; R.sup.3 is chloro or
bromo; R.sup.5 is hydroxy; and R.sup.4 and R.sup.6 are both
hydrogen.
[0040] Preferred benzthiazoles are those wherein R.sup.1 is
4-hydroxyphenyl; R.sup.6 is cyano or carboxy; R.sup.4 is hydroxy;
and R.sup.3 and R.sup.5 are both hydrogen.
[0041] Compounds within the formula (I) have been disclosed in the
literature: J. Med. Chem, 37 (1997) pages 1689-1695; British
Journal of Cancer, 77 (1998) pages 745-752; Chem. Pharm. Bull, 40
(1995) pages 2387-2390; EP483502, USP 5216110 and JP 2306916.
[0042] In another aspect the present invention provides compounds
of the formula (I) and pharmaceutically acceptable salts thereof
with the provisos that when X is S and:
a) R.sup.1 is 4-methoxyphenyl, the benzene ring of the benzthiazole
is not unsubstituted and is not substituted by 4-methyl,
4,6-dimethoxy, 5-methoxy, 5,6-dimethoxy, 6-methoxy, 6-chloro or
7-methoxy;
b) R.sup.1 is 3-methoxyphenyl, the benzene ring of the benzthiazole
is not unsubstituted and is not substituted by 6-methoxy;
c) R.sup.1 is 3,4-dimethoxyphenyl, the benzene ring of the
benzthiazole is not substituted by 6-methoxy, 4,6-dimethoxy or
5,6-dimethoxy;
d) R.sup.1 is phenyl, the benzene ring of the benzthiazole is not
substituted by 4-methoxy, 5,6-dimethoxy, 6-hydroxy or
6-methoxy;
e) R.sup.1 is 4-hydroxyphenyl, the benzene ring of the benzthiazole
is not unsubstituted and is not substituted by 4,6-dihydroxy,
5-hydroxy, 5,6-dihydroxy or 6-hydroxy;
f) R.sup.1 is 3,4-dihydroxyphenyl, the benzene ring of the
benzthiazole is not substituted by 6-hydroxy, 4,6-dihydroxy or
5,6-dihydroxy;
g) R.sup.1 is 2-hydroxyphenyl or 3-hydroxyphenyl, the benzene ring
of the benzthiazole is not substituted by 6-hydroxy;
h) R.sup.1 is 4-methylphenyl, the benzene ring of the benzthiazole
is not unsubstituted and is not substituted by 4-, 5- or 6-fluoro,
4-, 6- or 7-methoxy, 5-chloro, 4-, 5-, 6- or 7-hydroxy, 4-, 5-, 6-
or 7-acetoxy or 6-nitro;
i) R.sup.1 is 3,5-di-tert-butyl-4-hydroxyphenyl, the benzene ring
of the benztriazole is not substituted by 4- or 5-hydroxy;
and when X is S, R.sup.1 is not 4-aminophenyl,
4-amino-3-methylphenyl or 4-amino-3-halophenyl and when X is S or
O, R.sup.1 is not 4-chloro- or 4-fluorophenyl when the benzene ring
of the benzthiazole is substituted by 5-hydroxy or 5-mercapto.
[0043] Particular embodiments, particular aspects and preferred
features of the compounds of this invention are as described above
for the compounds for use in treating disease conditions related to
the .beta.-estrogen receptor.
[0044] Particularly useful compounds have any of the above
embodiments and also satisfy the equation:
(K.sub.i.alpha.A/K.sub.i.beta.A)/(K.sub.i.alpha.E/K.sub.i.beta.E)>100,
wherein
[0045] K.sub.i.alpha.A is the K.sub.i value for the agonist in
ER-.alpha.;
[0046] K.sub.i.beta.A is the K.sub.i value for the agonist in
ER-.beta.;
[0047] K.sub.i.alpha.E is the K.sub.i value for estrogen in
ER-.alpha.; and
[0048] K.sub.i.beta.E is the K.sub.i value for estrogen in
ER-.beta..
[0049] Another aspect of the invention is the use of any of the
above compound embodiments for the manufacture of a medicament for
the treatment or prophylaxis of Alzheimer's disease, anxiety
disorders, depressive disorders, osteoporosis, cardiovascular
disease, rheumatoid arthritis or prostate cancer.
[0050] Another aspect of the invention is the use of any of the
above compound embodiments in the treatment or prophylaxis of
Alzheimer's disease, anxiety disorders, depressive disorders
(including post-partum and post-menopausal depression),
osteoporosis, cardiovascular disease, rheumatoid arthritis or
prostate cancer.
[0051] C.sub.Y-Zalkyl, unless otherwise specified, means an alkyl
chain containing a minimum Y total carbon atoms and a maximum Z
total carbon atoms. These alkyl chains may be branched or
unbranched, cyclic, acyclic or a combination of cyclic and acyclic.
It also includes saturated and unsaturated alkyl such as ethynyl
and propenyl. For example, the following substituents would be
included in the general description "C.sub.4-7alkyl": ##STR4##
[0052] The term "oxo" means a double bonded oxygen (.dbd.O).
[0053] The compounds of the invention may contain heterocyclic
substituents that are 5- or 6-membered ring heterocycles containing
1, 2 or 3 heteroatoms each independently selected from O, N and S
and additionally having 0 or 1 oxo groups and 0 or 1 fused benzo
rings. A nonexclusive list containing specific examples of such
heterocycles are as follows: ##STR5## wherein the crossed bond
represents that the heterocycle may be attached at any available
position on either the heterocycle or the benzo ring.
[0054] Some of the compounds of the present invention are capable
of forming salts with various inorganic and organic acids and bases
and such salts are also within the scope of this invention.
Examples of such acid addition salts include acetate, adipate,
ascorbate, benzoate, benzenesulfonate, bisulfate, butyrate,
camphorate, camphorsulfonate, citrate, cyclohexyl sulfamate,
ethanesulfonate, fumarate, glutamate, glycolate, hemisulfate,
2-hydroxyethyl-sulfonate, heptanoate, hexanoate, hydrochloride,
hydrobromide, hydroiodide, hydroxy-maleate, lactate, malate,
maleate, methanesulfonate, 2-naphthalenesulfonate, nitrate,
oxalate, pamoate, persulfate, phenylacetate, phosphate, picrate,
pivalate, propionate, quinate, salicylate, stearate, succinate,
sulfamate, sulfanilate, sulfate, tartrate, tosylate
(p-toluene-sulfonate), and undecanoate. Base salts include ammonium
salts, alkali metal salts such as sodium, lithium and potassium
salts, alkaline earth metal salts such as aluminum, calcium and
magnesium salts, salts with organic bases such as dicyclohexylamine
salts, N-methyl-D-glucamine, and salts with amino acids such as
arginine, lysine, omithine, and so forth. Also, basic
nitrogen-containing groups may be quaternized with such agents as:
lower alkyl halides, such as methyl, ethyl, propyl, and butyl
halides; dialkyl sulfates like dimethyl, diethyl, dibutyl; diamyl
sulfates; long chain halides such as decyl, lauryl, myristyl and
stearyl halides; aralkyl halides like benzyl bromide and others.
Non-toxic physiologically-acceptable salts are preferred, although
other salts are also useful, such as in isolating or purifying the
product.
[0055] The salts may be formed by conventional means, such as by
reacting the free base form of the product with one or more
equivalents of the appropriate acid in a solvent or medium in which
the salt is insoluble, or in a solvent such as water, which is
removed in vacuo or by freeze drying or by exchanging the anions of
an existing salt for another anion on a suitable ion-exchange
resin.
Estrogen Receptor Binding Measurements
[0056] Abbreviated Procedure for Fluorescence Polarization Estrogen
Receptor (ERFP) Binding Assay
[0057] A homogeneous mix-and-measure estrogen receptor (ER) binding
assay which utilizes fluorescence polarization (FP) technology is
used to identify compounds with affinity for the estrogen receptor.
Purchased from PanVera (Madison, Wis.), assay reagents include
purified human recombinant ER.alpha., human recombinant ER.beta.,
ES2 screening buffer (100 mM potassium phosphate, pH 7.4, 100
.mu.g/mL bovine gamma globulin), and Fluormone.TM. ES2.
Fluormone.TM. ES2, whose formulation is proprietary to PanVera, is
a fluorescein-tagged, estrogen-like molecule which exhibits
approximately equal affinity for ER.alpha. and ER.beta..
[0058] For competition binding experiments, dilutions of test
compounds are prepared at 2.times. the final assay concentration in
0.2% DMSO in ES2 Screening buffer on TECAN Genosys, and 25 .mu.L
compound/well is dispensed into black Costar 1/2 volume 96-well
plates. Dependent upon a lot specific K.sub.d determination, 10-40
nM ER.alpha. or 1040 nM ER.beta. and 1 nM Fluormone ES2 are then
added to these plates in a final assay volume of 50 .mu.L/well.
Plates are gently shaken for at least 5 minutes to mix and
incubated for at least 1 hr 45 minutes to achieve equilibrium.
(Reaction mixtures are stable for up to 5 hours). After
centrifugation to remove air bubbles, plates are read on an LJL
Analyst or Acquest equipped with Criterion software at the
following settings: Fluorescence Polarization Mode; Static
Polarizer on Excitation Side; Dynamic Polarizer on Emission Side;
Excitation .lamda.=485+/-10 nm; Emission .lamda.=520+/-12.5 nm.
[0059] Polarized fluorescence intensity values are collected and
subsequently converted electronically to millipolarization (mp)
values. Following data reduction and normalization with Excel
and/or Prism software, % Ctrl values at the various test
concentrations are used to obtain IC.sub.50 values via non-linear
regression analysis of a four-parameter logistic equation.
[0060] Because ligand depletion is a consideration in this assay
(.about.40-60% input ES2 is bound in the assay), IC.sub.50 values
are converted to K.sub.i values through application of the Kenakin
formula, as outlined in the reference below, rather than via the
more routinely-used Cheng-Prusoff formula.
Reference: Bolger et al., Rapid Screening of Environmental
Chemicals for Estrogen Receptor Binding Capacity, Environmental
Health Pespectives: 106 (1998), 1-7.
Cell-based Assay for ER Transcriptional Activity:
[0061] ERs are ligand-dependent transcription factors that bind the
promoter regions of genes at a consensus DNA sequence called the
estrogen responsive element (ERE). The ER agonist or antagonist
activity of a drug was determined by measuring the amount of
reporter enzyme activity expressed from a plasmid under the control
of an estrogen-responsive element when cells transiently
transfected with ER and the reporter plasmid were exposed to drug.
These experiments were conducted according to the following
methods.
Plasmids:
[0062] Estrogen Receptors alpha (.alpha.ER, Gen Bank accession
#M12674), and beta (.beta.ER, Gen Bank # X99101 were cloned into
the expression vector pSG5 (Stratagene) and pcDNA3.1. A trimer of
the vitellogenin-gene estrogen response element (vitERE) was
synthesized as an oligonucleotide and attached to a beta-globin
basal promoter in a construct named pERE3gal. This response element
and promoter were removed from pERE3gal by digestion with the
endonucleases SpeI (filled with Klenow fragment) and HindIII. This
blunt/Hind III fragment was cloned into the .beta.-galactosidase
(.beta.-gal) enhancer reporter plasmid (pBGALenh, Stratagene).
.alpha.ER and .beta.ER plasmids were purified using a the Endo Free
Maxi Kit (Qiagen), and the DNA concentration and purity (A260/280
ratio) were determined spectrophotometrically (Pharmacia). Only DNA
with A260/280 ratio of 1.8 and a concentration of >1 ug/uL was
used for transfections.
[0063] Vitellogenin Response Element Sequence: TABLE-US-00001 (SEQ
ID NO: 1) CTAGTCTCGAGAGGTCACTGTGACCTAGGTCACTGTGACCTAGATCTAG
GTCACTGTGACCTAC =Spel overhang =Xhol site =Aflll overhang =ERE
consensus =spacer Bgl II
Cells:
[0064] All Transfections are performed in 293 cells (Human
Embryonic Kidney cells ATCC # CRL-1573). Cells are grown in DMEM
supplemented with 10% FBS, glutamine, sodium pyruvate and
penicillin/streptomycin. Cells are grown to 80% confluency and
split 1:10 or 1:20.
Transfection:
1. 293T cells are split the night before onto collagen 1150 mm
plates (Biocoat Becton Dickinson #354551) at 5 million cells per
plate in phenol red-free DMEM (Mediatech 17-205-CV) 10% FBS
charcoal stripped (biocell #6201-31) with supplements.
2. The next day the media is changed, 1 hour prior to transfection,
to fresh phenol red-free DMEM 10% FBS (charcoal stripped) and
supplements.
3. Transfections are performed using the Profection Kit from
Promega #El 200, this kit is based on calcium phosphate mediated
transfection. Reagents are added in sterile polystyrene tubes in
the following order:
[0065] Solution A
[0066] 20 ug ER alpha or beta (in pcDNA3.1)
[0067] 50 ug Reporter (pERE3 betaGal)
[0068] 1.5 ML Sterile Water
[0069] 186 uL CaCl.sub.2
[0070] * Mix gently
[0071] Solution B
[0072] 1.5 ml 2.times.HBSS
4. Using a vortex set on low, add solution A to solution B
dropwise. The resulting solution should become milky in color. It
is important to get thorough mixing at this point. Let solution
stand 30 min. Vortex before adding to cells.
[0073] 5. Add the mixture to 150-mm plates dropwise. Mix well by
rocking plates back and forth and side to side gently. View cells
under 20.times. magnification, a very fine precipitate should be
seen floating on and above cells after an hour. If you do not
observe this the transfection will not work well. Incubate 18-20
hours.
Receptor Stimulation:
[0074] 6. The day after transfection, cells are washed 2.times.
with PBS Ca Mg freePBS containing 1 mM EGTA pH=7.6. Cells are
trypsinized for 5 min with 4 ml of trypsin (0.25%)-EDTA. Trypsin is
neutralized with 6 ml DMEM (no phenol red)+10% charcoal stripped
FBS. Cells are pelleted at 1000.times.g for 5 min. Cell pellet is
resuspended in 10 ml DMEM (no phenol red)+2% charcoal stripped FBS
supplemented with glutamine and Penn/Strep and the cells are
counted. Additional medium is added to dilute the cell density to
500,000 cells/ml.
7. Cells are plated into 96 well dish (Biocoat BD #354407) at 50 ul
of cells per well (=25,000 cells/well), using a multichannel
pipettor. Plates are incubated for approx. 2-4 hours to allow cells
to attach.
[0075] 8. Compounds are prepared at concentration of 4 mM in 100%
DMSO, then diluted into medium with supplements but no serum. The
first 2 dilutions are done in medium with no DMSO, then the
remaining dilutions are in medium plus 0.5% DMSO to keep the
vehicle constant. Max controls are 10 nM beta-estradiol and
background controls are 0.5% DMSO. Compounds are normally tested in
the range of 10 uM to 1 nM and are prepared at twice the
concentration to be tested. The compounds are added to the cell
plates, 50 ul per well. All compounds are tested with an n=4 wells
for single poke and n=2 for 9-pt curves.
9. Cells are incubated overnight at 37.degree. C. with the
compounds.
[0076] Reporter Assay:
[0077] 1. After 18-24 hr of stimulation, 100 ul of 7% CPRG cocktail
is added to each well, the plate is incubated at 37C for
approximately 30 minutes to 2 hours or until the OD reaches between
1.0 and 2.0. The CPRG (Roche 0884308) will turn bright red as Beta
Gal cleaves it.
[0078] 2. The plates are read on a spectrophotometric plate reader
(Spectramax, Molecular Devices) at 570 nm and raw absorbances are
obtained.
Data is compiled and interpreted with Excel using XLFit or GraphPad
Prism to fit concentration-response curves. The EC50 is defined as
the concentration at which 50% of the fitted maximum for a compound
has been reached.
10.times. Z Buffer
[0079] TABLE-US-00002 Sodium Phosphate (dibasic) 1.7 g 600 mM
Sodium Phosphate (monobasic) 0.96 g 400 mM Potassium Chloride 149
mg 100 mM Magnesium Sulfate 0.2 mL of 1 molar stock 100 mM BME 0.78
mL 500 mM Bring Final Volume to 20 mL with De-Ionized Water
7% CPRG Cocktail
For 50 mLs:
add 3.5 mL of 50 ml of CPRG
add 3.5 mL of 10.times. Z Buffer
add 1 mL of 10% SDS
bring to 50 mL with DI water
Typical Results:
Absorbance values illustrating typical concentration-response
curves obtained for the ER agonist 17-.beta.-estradiol (E) and the
ER antagonist ICI182,780 (A) are plotted for cells transfected with
either .alpha.ER or .beta.ER (see FIGS. 1 and 2).
Administration and Use
[0080] Compounds of the present invention are shown to have high
selectivity for ER-.beta. over ER-.alpha., and may possess agonist
activity on ER-.beta. without undesired uterine effects. Thus,
these compounds, and compositions containing them, may be used as
therapeutic agents in the treatment of various CNS diseases related
to ER-.beta., such as, for example, Alzheimer's disease.
[0081] The present invention also provides compositions comprising
an effective amount of compounds of the present invention,
including the nontoxic addition salts, amides and esters thereof,
which may, serve to provide the above-recited therapeutic benefits.
Such compositions may also be provided together with
physiologically-tolerable liquid, gel or solid diluents, adjuvants
and excipients. The compounds of the present invention may also be
combined with other compounds known to be used as therapeutic
agents for the above or other indications.
[0082] These compounds and compositions may be administered by
qualified health care professionals to humans in a manner similar
to other therapeutic agents and, additionally, to other mammals for
veterinary use, such as with domestic animals. Typically, such
compositions are prepared as injectables, either as liquid
solutions or suspensions; solid forms suitable for solution in, or
suspension in, liquid prior to injection may also be prepared. The
preparation may also be emulsified. The active ingredient is often
mixed with diluents or excipients which are physiologically
tolerable and compatible with the active ingredient. Suitable
diluents and excipients are, for example, water, saline, dextrose,
glycerol, or the like, and combinations thereof. In addition, if
desired the compositions may contain minor amounts of auxiliary
substances such as wetting or emulsifying agents, stabilizing or
pH-buffering agents, and the like.
[0083] The compositions are conventionally administered
parenterally, by injection, for example, either subcutaneously or
intravenously. Additional formulations which are suitable for other
modes of administration include suppositories, intranasal aerosols,
and, in some cases, oral formulations. For suppositories,
traditional binders and excipients may include, for example,
polyalkylene glycols or triglycerides; such suppositories may be
formed from mixtures containing the active ingredient. Oral
formulations include such normally employed excipients as, for
example, pharmaceutical grades of mannitol, lactose, starch,
magnesium stearate, sodium saccharin, cellulose, magnesium
carbonate, and the like. These compositions take the form of
solutions, suspensions, tablets, pills, capsules, sustained-release
formulations, or powders.
[0084] In addition to the compounds of the present invention that
display ER-.beta. activity, compounds of the present invention can
also be employed as intermediates in the synthesis of such useful
compounds.
Synthesis
[0085] Compounds within the scope of the present invention may be
synthesized chemically by means well known in the art. The
following Examples are meant to show general synthetic schemes,
which may be used to produce many different variations by employing
various commercially available starting materials. These Examples
are meant only as guides on how to make some compounds within the
scope of the invention, and should not be interpreted as limiting
the scope of the invention.
In another aspect, the present invention provides a process for
preparing the compounds of the formula (I) and pharmaceutically
acceptable salts thereof, which comprises:
[0086] a) cyclising a compound of the formula: ##STR6## wherein X,
R.sup.1, R.sup.3-R.sup.6 are as defined hereinabove and L is
hydrogen or a leaving group; or b) for preparing compounds wherein
X is O, cyclising a compound of the formula: ##STR7## wherein
R.sup.1, R.sup.3-R.sup.6 are as defined hereinabove or: c)
cyclising a compound of the formula: ##STR8## wherein X, R.sup.1,
R.sup.3-R.sup.6 are as defined hereinabove and L is hydrogen or a
leaving group; or d) for preparing compounds wherein X is S,
cyclising a compound of the formula: ##STR9## wherein R.sup.1,
R.sup.3-R.sup.6 are as defined hereinabove; and thereafter, if
necessary:
[0087] i) forming a pharmaceutically acceptable salt;
[0088] ii) converting a compound of the formula (I) into another
compound of the formula (I).
EXAMPLES
[0089] TABLE-US-00003 Synthetic HPLC MS Example Structure Method
(min) (MH.sup.+) 1 ##STR10## A, B, C 1.99 262 2 ##STR11## A, B 276
3 ##STR12## B, C 242 4 ##STR13## B 256 5 ##STR14## 244 6 ##STR15##
B 228 7 ##STR16## B, C 226 (M-H.sup.-) 8 ##STR17## B 242 9
##STR18## D, E, F, G, H, I 2.32 296 10 ##STR19## F, G, H, I, J 2.29
308 11 ##STR20## F, G, H, I, J 2.18 321 12 ##STR21## F, G, H, I, J
2.30 341 13 ##STR22## F, G, H, I, J, K 2.26 354 14 ##STR23## D, E,
G, H, I 2.07 262 15 ##STR24## G, H, I 2.03 242 16 ##STR25## Ref. 1,
F, G, H, I 2.25 256 17 ##STR26## Ref. 1, F, G, H, I 2.36 276 18
##STR27## Ref. 1, D, E, F, G, H, I 2.02 276 19 ##STR28## D, E, F,
G, H, I 2.17 278 20 ##STR29## F, G, H, I 1.82 258 21 ##STR30## J,
F, G, H, K, L, I 2.89 290 22 ##STR31## J, F, G, H, M, I 253 2.06 23
##STR32## J, F, G, H, I, N 2.04 271 24 ##STR33## D, E, J, F, G, H,
I, 2.30 341 25 ##STR34## J, F, G, H, I 2.32 322 26 ##STR35## D, E,
J, F, G, H, K, I 2.24 388 27 ##STR36## J, F, G, H, K, I 2.22 368 28
##STR37## D, E, J, F, G, H, I, M 2.05 287 29 ##STR38## J, F, G, H,
K, I, M 2.02 267 30 ##STR39## D, E, J, F, G, H, I, O 1.92 278 31
##STR40## P, F, G, H, I 2.18 308 32 ##STR41## Q, R, I 1.20 237 33
##STR42## C 2.44 290.3, 292.3 34 ##STR43## J, F, Q, R, I 2.69 359
35 ##STR44## B 1.80 252.4 36 ##STR45## 1.84 352.2, 354 (90%)
(MH.sup.++CH.sub.3CN); 311.2, 313 (20%) (MH.sup.+)
[0090] Biological Data for Representative Compounds of this
Invention: TABLE-US-00004 Example FP .beta.-ER K.sub.i (nM) FP
.alpha.-ER K.sub.i (nM) FP Selectivity 1 1.7 18 10 3 4.8 121 25 5
290 1000 3 7 5.8 82 14 8 148 477 3 10 0.38 5.6 15 12 1.8 54 30 22
1.2 14 12 23 646 2200 3
[0091] TABLE-US-00005 ERE ERE .beta.-ER ERE .beta.-ER ERE
.alpha.-ER ERE .alpha.-ER Selec- Example EC.sub.50 (nM) Max
EC.sub.50 (nM) Max tivity 1 1.2 98 61 103 52 3 23 89 497 102 22 5
1000 98 1000 102 1.8 7 1.4 95 25 116 18 8 10 0.017 103 6.0 109 363
12 0.5 81 76 37 153 22 0.616 98 6.4 83 10 23 298 86 1000 78 14
Chemical Syntheses The HPLC conditions used are the following
unless stated otherwise: HPLC 2.1.times.50 mm C.sub.8 5 .mu.m
Zorbax Stablebond column; flow rate 1.4 mL/min, linear gradient
from 15% B to 90% B over 4.0 min; A=water, 0.05% TFA; B=90%
acetonitrile, 10% water, 0.05% TFA, UV detection at 254 nm or DAD
and positive ionization mass spectrometry detection.
[0092] TFA: trifluoroacetic acid
[0093] DMSO: dimethylsulfoxide
[0094] DEAD: diethyl axodicarboxylate
[0095] PPh.sub.3: triphenylphosphine
[0096] EDTA: ethylenediaminetetraacetic acid
[0097] BBr.sub.3: boron tribromide
Example 1
4-Chloro-6-hydroxy-2-(4-hydroxyphenyl)benzoxazole
1) Synthetic method A: Synthesis of
2-amino-3-chloro-5-methoxyphenol
[0098] A solution of 3-chloro-5-methoxy-2-nitrophenol [1] (450 mg)
and tin(II) chloride dihydrate (2 g, 4 eq.) in ethyl acetate (30
mL) was heated under reflux for 4 h. The mixture was cooled,
diluted with ethyl acetate and aqueous potassium fluoride. The
mixture was filtered through celite. The organic layer was washed
with brine and dried over MgSO.sub.4. Evaporation of the solvent
afforded the title compound (280 mg) as a pale solid. NMR
(DMSO-d.sub.6): 9.74 (m, 1H), 6.33 (d, 1H, J=2.4 Hz), 6.30 (d, 1H,
J=2.4 Hz), 4.19 (m, 2H), 3.60 (s, 3H).
[0099] Reference 1: Hodgson, Wignall, J. Chem. Soc., 1928, 330.
Prepared from 1-chloro-3,5-dimethoxybenzene by nitration with
concentrated nitric acid in acetic anhydride below 10.degree. C. to
give 1-chloro-3,5-dimethoxy-2-nitrobenzene, and subsequent reaction
with boron tribromide (1 eq.) in dichloromethane from -78.degree.
C. to 0.degree. C.
2) Synthetic method B: Synthesis of
4-chloro-2-(4-hydroxyphenyl)-6-methoxybenzoxazole
Example 2
[0100] A solution of 2-amino-3-chloro-5-methoxyphenol (270 mg) and
ethyl 4-hydroxybenzimidate hydrochloride (376 mg, 1.2 eq.) in
absolute ethanol (5 mL) was heated under reflux for 4 h. The
mixture was cooled, partitioned between ethyl acetate and water.
The organic layer was dried over MgSO.sub.4. After evaporation of
the solvent, the residue was triturated with methanol to give the
title compound (130 mg) as a light orange solid. NMR
(DMSO-d.sub.6): 10.35 (s, 1H), 8.00 (d, 2H, J=8.7 Hz), 7.39 (d, 1H,
J=2.1 Hz), 7.10 (d, 1H, J=2.1 Hz), 6.96 (d, 2H, J=8.7 Hz), 3.85 (s,
3H); MS: 276 (MH.sup.+).
3) Synthetic method C: Synthesis of
4-chloro-6-hydroxy-2-(4-hydroxyphenyl)benzoxazole
[0101] To a suspension of
4-chloro-2-(4-hydroxyphenyl)-6-methoxybenzoxazole (240 mg) in
dichloromethane (5 mL) cooled at -78.degree. C. was added boron
tribromide (5 mL, 1M solution in dichloromethane, 5.7 eq.). The
mixture was stirred at -78.degree. C. for 10 min, warmed to room
temperature and stirred for 3 h. The mixture was poured onto
ice/water and extracted with ethyl acetate. The organic layer was
washed with brine and dried over MgSO.sub.4. Chromatography on a
silica gel column (eluant: acetone -dichloromethane, gradient from
0:100 to 5:95) and trituration of the resulting solid in
dichloromethane afforded the title compound (53 mg) as a solid. NMR
(DMSO-d.sub.6): 10.31 (s, 1H), 10.17 (s, 1H), 7.98 (d, 2H, J=8.4
Hz), 7.06 (s, 1H), 6.95 (d, 2H, J=8.4 Hz), 6.87 (s, 1H); MS: 262
(MH.sup.+); HPLC t.sub.R: 1.99 min.
Example 3
6-Hydroxy-2-(4-hydroxyphenyl)-4-methylbenzoxazole
1) Synthesis of 2-(4-hydroxyphenyl)-6-methoxy-4-methylbenzoxazole
(Example 4)
According to synthetic method B, from
2-amino-5-methoxy-3-methylphenol [2] (440 mg) was obtained the
title compound (340 mg) as a light orange solid. MS: 256
(MH.sup.+).
[0102] Reference 2: Musso H; Beecken H, Chem. Ber. 1961, 94, 585;
made from 3,5-dimethoxytoluene by nitration and monodeprotection of
the 3-methoxy with BBr.sub.3 similarly to Ref. 1 followed by
reduction of the nitro group to the amino by hydrogenation with
palladium on charcoal.
[0103] 2) According to synthetic method C, the above compound (220
mg) was converted to
6-hydroxy-2-(4-hydroxyphenyl)-4-methylbenzoxazole (112 mg) as a
light pinkish powder. NMR (DMSO-d.sub.6): 10.14 (s, 1H), 9.58 (s,
1H), 7.94 (d, 2H, J=8.7 Hz), 6.92 (d, 2H, J=8.7 Hz), 6.84 (s, 1H),
6.63 (s, 1H), 2.46 (s, 3H); MS: 242 (MH.sup.+).
Example 5
4,6-Dihydroxy-2-(4-hydroxyphenyl)benzoxazole
[0104] A mixture of 1-nitro-2,4,6-trihydroxybenzene (1 g) and 10%
palladium on charcoal (200 mg) in absolute ethanol (10 mL) was
stirred for 18 h at room temperature under a 50-PSI atmosphere of
hydrogen. The solids were filtered rapidly. To the resulting
filtrate was added ethyl 4-hydroxybenzimidate hydrochloride (1.17
g). The mixture was heated at reflux for 5 h under nitrogen, cooled
and partitioned between ethyl acetate and water. The organic layer
was washed with water and brine, and dried over MgSO4.
Chromatography on silica gel (eluant: acetone-dichloromethane,
gradient 10:90 to 20:80) and trituration of the resulting solid
with ether afforded the title compound (28 mg) as a light pinkish
solid. NMR (DMSO-d.sub.6): 10.15 (m, 2H), 9.53 (s, 1H), 7.91 (d,
2H, J=8.7 Hz), 6.92 (d, 2H, J=8.7 Hz), 6.48 (d, 1H, J=1.5 Hz), 6.26
(d, 1H, J=1.5 Hz); MS: 244 (MH.sup.+).
Example 6
6-Hydroxy-2-(4-hydroxyphenyl)benzoxazole
[0105] According to synthetic method B except that pyridine (300
.mu.L) was added, from 4-aminoresorcinol hydrochloride (435 mg) and
ethyl 4-hydroxybenzimidate hydrochloride was obtained the title
compound (432 mg) as an off-white solid. The work-up of the
reaction was modified as follows: After completion of the reaction,
the mixture was diluted with ether and water, and filtered. The
solids were washed with water and ether, and dried under high
vacuum. NMR (DMSO-d.sub.6): 10.23 (s br, 1H), 9.76 (s br, 1H), 7.95
(d, 2H, J=8.4 Hz), 7.50 (d, 1H, J=8.4 Hz), 7.05 (d, 1H, J=1.8 Hz),
6.94 (d, 2H, J=8.4 Hz), 6.81 (dd, 1H, J=8.4 Hz, J'=1.8 Hz); MS: 228
(MH.sup.+).
Example 7
5-Hydroxy-2-(4-hydroxyphenyl)benzoxazole
[0106] According to synthetic method B, from
2-amino-4-methoxyphenol [3] (500 mg) and ethyl 4-hydroxybenzimidate
hydrochloride was obtained 2-(4-hydroxyphenyl)-5-methoxybenzoxazole
(676 mg; Example 8). NMR (DMSO-d.sub.6): 10.28 (s, 1H), 8.01 (d,
2H, J=8.7 Hz), 7.61 (d, 1H, J=9 Hz), 7.28 (d, 1H, J=2.4 Hz), 6.94
(m, 3H), 3.82 (s, 3H); MS: 242 (MH.sup.+). This compound (452 mg)
was converted to the title compound (121 mg) according to synthetic
method C. NMR (DMSO-d.sub.6): 10.24 (s, 1H), 9.42 (s br, 1H), 7.99
(d, 2H, J=8.7 Hz), 7.49 (d, 1H, J=9 Hz), 7.02 (d, 1H, J=2.1 Hz),
6.94 (d, 2H, J=8.7 Hz), 6.78 (dd, 1H, J=9 Hz, J'=2.1 Hz); MS: 226
(M-H.sup.-). Reference 3: Lok R, Leone R E, Williams A J, J. Org.
Chem. 1996, 61, 3289. ##STR46##
Example 9
4-Chloro-2-(2-chloro-4-hydroxy-phenyl)-benzooxazol-6-ol
Synthetic method D: Synthesis of 2-chloro-4-methoxy-benzoic acid
methyl ester
[0107] To 2-chloro-4-hydroxy-benzoic acid hydrate (1.1 g) in DMF
(20 mL) was added K.sub.2CO.sub.3 (2.4 g) and methyl iodide (0.75
mL). After 2 h, water was added to the reaction mixture and
extracted with ethyl acetate. The organic layer was washed with
brine (3.times.) and dried over MgSO.sub.4. Flash chromatography on
silica gel eluting with 10% ethyl acetate-hexane afforded 1.13 g
(95%) of the title compound as an oil. MS: 201 (MH.sup.+), HPLC
t.sub.R: 2.52 min.
Synthetic method E: Synthesis of 2-chloro-4-methoxy-benzoic
acid
[0108] To 2-chloro-4-methoxy-benzoic acid methyl ester (1.1 g) in
THF/MeOH/water (12 mL/3 mL/3 mL) at room temperature was added LiOH
(461 mg) dissolved in water. After 2 h, the reaction mixture was
adjusted to pH 4 with 1N HCl and partitioned between ethyl acetate
and water. The organic layer was washed with brine and dried over
MgSO.sub.4. Trituration of the resulting solid in ether afforded
1.0 g (98%) of the title compound. MS: 187 (MH.sup.+), HPLC
t.sub.R: 2.04 min.
Synthetic method F: Synthesis of
2-amino-3-chloro-5-methoxyphenol
[0109] To 3-chloro-5-methoxy-2-nitrophenol, synthetic method A [1],
(200 mg) in 95% EtOH (10 mL) was added 5% Ru/C (20 mg) and
hydrazine (0.36 mL). The mixture was placed in an oil bath and
heated to 85.degree. C. for 2 h. After reaction cooled, the mixture
was diluted with ethyl acetate and filtered through a pad of celite
and concentrated. Flash chromatography on silica gel eluting with
30% ethyl acetate-hexane afforded 138 mg (76%) of the title
compound as a solid. MS: 174 (MH.sup.+), HPLC t.sub.R: 0.84
min.
Synthetic method G: Synthesis of
2-chloro-N-(2-chloro-6-hydroxy-4-methoxy-phenyl)-4-methoxy-benzamide.
[0110] To 2-chloro-4-methoxy-benzoic acid (100 mg) in
CH.sub.2Cl.sub.2 (5 mL) was added oxalyl chloride (0.05 mL) and 2
drops of DMF. The reaction mixture was allowed to stir for 2 h and
then concentrated to dryness. The resulting acid chloride was taken
up in CH.sub.2Cl.sub.2 and added drop wise to a cool mixture of
2-amino-3-chloro-5-methoxyphenol (94 mg), 10% Na.sub.2CO.sub.3 (2.5
mL), and CH.sub.2Cl.sub.2 (5 mL) placed in an ice water bath. After
2 h, water was added to the reaction and extracted with additional
CH.sub.2Cl.sub.2 and the organic layer was dried over
Na.sub.2SO.sub.4. Flash chromatography on silica gel eluting with
0% to 30% ethyl acetate-hexane afforded 100 mg (54%) of the title
compound as an solid. MS: 342 (MH.sup.+), HPLC t.sub.R: 2.54
min.
Synthetic method H: Synthesis of
4-chloro-2-(2-chloro-4-methoxy-phenyl)-6-methoxy-benzooxazole
Reference 4. Wang, F.; Hauske, J. R.; Tetrahedron Lett. 1997, 38
(37) 6529-6532
[0111]
2-Chloro-N-(2-chloro-6-hydroxy-4-methoxy-phenyl)-4-methoxy-benzami-
de (40 mg) was completely dissolved in THF (5 mL) and PPH.sub.3 (46
mg) was added and the mixture was stirred until all PPH.sub.3
dissolved. To this mixture was added drop wise DEAD (0.03 mL)
diluted in THF (0.5 mL). The mixture was stirred at room
temperature for 2 h and the reaction mixture was diluted with ethyl
acetate and washed with water, brine, dried over MgSO.sub.4 and
concentrated. Flash chromatography on silica gel eluting with 10%
to 30% ethyl acetate-hexane afforded 32 mg (85%) of the title
compound as a solid. MS: 324 (MH.sup.+), HPLC t.sub.R: 3.18
min.
Synthetic method I: Synthesis of
4-chloro-2-(2-chloro-4-hydroxy-phenyl)-benzooxazol-6-ol
[0112] To
4-chloro-2-(2-chloro-4-methoxy-phenyl)-6-methoxy-benzooxazole (52
mg) in CH.sub.2Cl.sub.2 (2 mL) placed in and ice water bath was
added drop wise 1.0 M BBr.sub.3 in CH.sub.2Cl.sub.2 (0.96 mL, 6
eq). After the reaction stirred over night, the mixture was placed
in an ice water bath and excess MeOH was added drop wise to quench
excess BBr.sub.3 and the mixture was stirred for an additional 20
min and concentrated. Flash chromatography on silica gel eluting
with 0% to 40% ethyl acetate-hexane afforded 35 mg (74%) of the.
title compound as a solid. MS: 296 (MH.sup.+), HPLC t.sub.R: 2.32
min.
Example 10
7-Bromo-2-(4-hydroxy-phenyl)-benzooxazol-5-ol
Synthetic method J: Synthesis of
2-bromo-4-methoxy-6-nitro-phenol
[0113] 4-Methoxy-2-nitro-phenol (10 g) was dissolved in glacial
acetic acid (60 mL) and CH.sub.3CO.sub.2Na (8.2 g) was added. Next,
bromine (3 mL) dissolved in glacial acetic acid (12 mL) was added
drop wise to the stirring solution at room temperature. After
complete addition of bromine, the mixture was stirred for 30 min at
room temperature and then placed in an oil bath at 75.degree. C.
for 2 h. After reaction mixture cooled to room temperature,
concentrated HCl (500 mL) was slowly added to the mixture followed
by addition of ethyl acetate (500 mL). The layers were separated
and the organic layer was washed with water, brine, dried
(Na.sub.2SO.sub.4). Flash chromatography on silica gel eluting with
5% ethyl acetate-hexane afforded 8.8 g (60%) of the title compound
as a solid. MS: 218 (MH.sup.+-30), HPLC t.sub.R: 2.40 min.
[0114] According to synthetic methods F, G, H (except the reaction
mixture was heated in an oil bath at 85.degree. C. for 2 h), and I
was obtained 7-bromo-2-(4-hydroxy-phenyl)-benzooxazol-5-ol. MS: 308
(MH.sup.+), HPLC t.sub.R: 2.29 min. NMR (DMSO-d.sub.6): 10.34 (s,
1H), 9.82 (s, 1H), 7.99 (d, 2H, J=8.4 Hz), 6.95-7.04 (m, 4H).
Example 12
7-Bromo-2-(3-chloro-4-hydroxy-phenyl)-benzooxazol-5-ol
[0115] According to synthetic methods J (using
4-methoxy-2-nitro-phenol), F, G (using 3-chloro-4-methoxy-benzoic
acid), H (except the reaction mixture was heated in an oil bath at
85.degree. C. for 2 h), and I the title compound was obtained. MS:
341 (MH.sup.+), HPLC t.sub.R: 2.30 min. NMR (DMSO-d.sub.6): 11.17
(s, 1H), 9.85 (s, 1H), 8.04 (s, 1H), 7.95 (d, 1H, J=8.2 Hz), 7.18
(d, 1H, J=8.3 Hz), 7.05 (s, 1H), 7.02 (s, 1H).
Example 13
2-(4-Hydroxy-phenyl)-7-iodo-benzooxazol-5-ol
Synthetic method K: Synthesis of
7-iodo-5-methoxy-2-(4-methoxy-phenyl)-benzooxazole
[0116] To 7-bromo-5-methoxy-2-(4-methoxy-phenyl)-benzooxazole (100
mg, from example 10, synthetic methods F, G, H, J), was added CuI
(285 mg), KI (497 mg) and DMSO (5 mL). The mixture was placed in an
oil bath and heated to 180.degree. C. for 4 h. The mixture was
cooled to room temperature and diluted with ethyl acetate and
washed with brine (3.times.), dried (MgSO.sub.4), and concentrated.
The crude solid was taken up in ethyl acetate and filtered through
a pad of celite and the filtrate was concentrated to dryness. Flash
chromatography on silica gel eluting with 20% ethyl acetate-hexane
afforded 80 mg (70%) of the title compound as a solid. MS: 382
(MH.sup.+), HPLC t.sub.R: 3.00 min. According to synthetic method I
was obtained 2-(4-Hydroxy-phenyl)-7-iodo-benzooxazol-5-ol. MS: 354
(MH.sup.+), HPLC t.sub.R: 2.26 min.
Example 21
7-Chloro-2-(4-hydroxy-phenyl)-benzooxazol-5-ol
Synthetic method L: Synthesis of
7-chloro-5-methoxy-2-(4-methoxy-phenyl)-benzooxazole
[0117] To 7-iodo-5-methoxy-2-(4-methoxy-phenyl)-benzooxazole (150
mg) in DMF (6 mL) was added CuCl (195 mg). The mixture was placed
in an oil bath and heated to 150.degree. C. for 3 h. The mixture
was cooled to room temperature and diluted with ethyl acetate and
washed with 1N HCl, brine (3.times.), dried (MgSO.sub.4), and
concentrated. Flash chromatography on silica gel eluting with 20%
ethyl acetate-hexane afforded 100 mg (88%) of the title compound as
a solid. MS: 290 (MH.sup.+), HPLC t.sub.R: 2.89 min.
According to synthetic method I was obtained
7-chloro-2-(4-hydroxy-phenyl)-benzooxazol-5-ol. MS: 262 (MH.sup.+),
HPLC t.sub.R: 2.09 min.
Example 22
5-Hydroxy-2-(4-hydroxy-phenyl)-benzooxazole-7-carbonitrile
Synthetic method M: Synthesis of
5-hydroxy-2-(4-hydroxy-phenyl)-benzooxazole-7-carbonitrile
[0118] To 7-bromo-2-(4-hydroxy-phenyl)-benzooxazol-5-ol (96 mg) in
DMF (3 mL) was added CuCN (84 mg). The mixture was placed in an oil
bath and heated to 150.degree. C. for 3 h. The mixture was cooled
to room temperature and diluted with ethyl acetate and washed with
1N HCl, saturated aqueous EDTA, brine (3.times.), dried
(Na.sub.2SO.sub.4), and concentrated. Flash chromatography on
silica gel eluting with 20% ethyl acetate-hexane afforded 20 mg
(25%) of the title compound as a solid. MS: 253 (MH.sup.+), HPLC
t.sub.R: 2.06 min.
Example 23
5-Hydroxy-2-(4-hydroxy-phenyl)-benzooxazole-7-carboxylic acid
amide
Synthetic method N: Synthesis of
5-methoxy-2-(4-methoxy-phenyl)-benzooxazole-7-carbonitrile
[0119] To 7-bromo-5-methoxy-2-(4-methoxy-phenyl)-benzooxazole (200
mg) in DMF (5 mL) was added CuCN (80 mg). The mixture was placed in
an oil bath and heated to 150.degree. C. for 3 h. The mixture was
cooled to room temperature and diluted with ethyl acetate and
washed with 1N HCl, brine (3.times.), dried (Na.sub.2SO.sub.4), and
concentrated. Flash chromatography on silica gel eluting with 20%
ethyl acetate-hexane afforded 85 mg (50%) of the title compound as
a solid. MS: 281 (MH.sup.+), HPLC t.sub.R: 2.71 min.
[0120] According to synthetic method I was obtained
5-hydroxy-2-(4-hydroxy-phenyl)-benzooxazole-7-carboxylic acid amide
by residual acid hydrolysis of the nitrile substituent upon
concentration of the crude reaction mixture. MS: 271 (MH.sup.+),
HPLC t.sub.R: 2.04 min.
Example 30
2-(2-Cyano-4-hydroxy-phenyl)-5-hydroxy-benzooxazole-7-carbonitrile
Synthetic method O: Synthesis of
2-(2-cyano-4-hydroxy-phenyl)-5-hydroxy-benzooxazole-7-carbonitrile
[0121] To 2-(2-chloro-4-hydroxy-phenyl)-7-iodo-benzooxazol-5-ol
(279 mg) in DMF (5 mL) was added CuCN (97 mg). The mixture was
placed in an oil bath and heated to 150.degree. C. for 3 h. The
mixture was cooled to room temperature and diluted with ethyl
acetate and washed with 1N HCl, saturated aqueous EDTA, brine
(3.times.), dried (Na.sub.2SO.sub.4), and concentrated. Material
was purified by preparative LC/MS. MS: 278 (MH.sup.+), HPLC
t.sub.R: 1.92 min. The HPLC conditions used are the following: HPLC
Waters Corp. Novapak HR.TM. C 18 RCM 40.times.100 mm 6 .mu.m
particle; flow rate 40 mL/min, linear gradient from 35% B to 65% B
over 15 min; A=water, 0.1% TFA; B=MeOH, UV detection at 254 nm and
positive ionization mass spectrometry detection
Example 31
6-Bromo-2-(4-hydroxy-phenyl)-benzooxazol-5-ol
Synthetic method P: Synthesis of 1
-bromo-2,5-dimethoxy-4-nitro-benzene
[0122] The title compound was synthesized according to the methods
describe by Reference 5: Jean-Luc Grenier, Jean-Pierre Catteau and
Philippe Cotelle, Synthetic Communications, 29(7), 1201-1208
(1999).
According to synthetic methods F, G, H and I, was obtained
6-bromo-2-(4-hydroxy-phenyl)-benzooxazol-5-ol. MS: 308 (MH.sup.+),
HPLC t.sub.R: 2.18 min.
Example 32
4-(6-Hydroxy-benzooxazol-2-yl)-thiazolidin-2-one
Reference 6. Steven W. Goldstien and Paul J. Dambek, J.
Heterocyclic Chem. 1990, 27, 225.
Synthetic method Q: Synthesis of 2-oxo-thiazolidine-4-carboxylic
acid (2-hydroxy-4-methoxy-phenyl)-amide
[0123] To 2-oxo-thiazolidine-4-carboxylic acid (492 mg) in
CH.sub.2Cl.sub.2 (10 mL) was added oxalyl chloride (0.35 mL) and 2
drops of DMF. The reaction mixture was allowed to stir for 2 h and
then concentrated to dryness. The resulting acid chloride was taken
up in CH.sub.2Cl.sub.2 and added drop wise to a cool mixture of
2-amino-5-methoxy-phenol (490 mg), Et.sub.3N (1.56 mL), and
CH.sub.2Cl.sub.2 (10 mL) placed in an ice water bath. After 2 h,
water and CH.sub.2Cl.sub.2 was added to the reaction mixture and
layers separated. The organic layer was washed with 1N HCl. To the
HCl aqueous layer was added brine and CH.sub.2Cl.sub.2 to salt-out
the title compound, the organic layers were combined, dried
(Na.sub.2SO.sub.4) and concentrated to afford 448 mg (60%) of the
title compound. The crude amide was used directly in the next step.
MS: 269 (MH.sup.+), HPLC t.sub.R: 1.37 min.
Synthetic method R: Synthesis of
4-(6-methoxy-benzooxazol-2-yl)-thiazolidin-2-one
[0124] To 2-oxo-thiazolidine-4-carboxylic acid
(2-hydroxy-4-methoxy-phenyl)-amide (50 mg) in p-xylene (3 mL) was
added pyridinium p-toluenesolfonate (42 mg). The reaction mixture
was refluxed for 2 h and then allowed to cool to room temperature.
The mixture was diluted with ethyl acetate and water was added. The
layers were separated and the organic layer was washed with brine,
dried (Na.sub.2SO.sub.4), and concentrated. Flash chromatography on
silica gel eluting with 3% methanol-methylene chloride afforded 42
mg (90%) of the title compound as a solid. MS: 251 (MH.sup.+), HPLC
t.sub.R: 1.60 min.
According to synthetic method I was obtained
4-(6-hydroxy-benzooxazol-2-yl)-thiazolidin-2-one 30 mg (63%). MS:
237 (MH.sup.+), HPLC t.sub.R: 1.20 min.
Example 33
7-Bromo-2-phenyl-benzooxazol-5-ol
[0125] tert-Butyl-chloro-silane (0.25 mL) was added to a mixture of
2-bromo-4-methoxy-6-nitro-phenol (0.124 g), chromium (II) chloride
(0.012 g) and manganese (0) powder (0.137 g) in dimethylformamide
(3 mL). The mixture was subjected to microwaves for 4 min at
150.degree. C. Benzaldehyde (0.06 mL) was added and the reaction
was subjected to microwaves for 6 min at 150.degree. C. Water (0.5
mL) was added, then after 30 min the mixture was filtered through a
pad of celite. The above procedure was repeated three more times.
The combined filtrate was partitioned between dilute aq. HCl (100
mL) and ethyl acetate (100 mL). The organic layer was dried over
sodium sulfate, filtered through celite and concentrated.
Chromatography on silica gel (eluant: gradient 0 to 30% ethyl
acetate:hexane) afforded 7-bromo-5-methoxy-2-phenyl-benzooxazole
(0.13 g). MS: 304 (95%), 306.4 (100%) (MH.sup.+); HPLC t.sub.R:
2.79 min.
[0126] Reference 7: J. Org. Chem. 2001, 66, 991-996.
7-Bromo-5-methoxy-2-phenyl-benzooxazole was deprotected using
synthetic method C to give the title compound (0.060 g) after
purification on silica gel (eluant: 5 to 30% ethyl acetate:hexane).
MS: 290.3, 292.3 (MH.sup.+); HPLC t.sub.R: 2.44 min.
Example 35
2-(1H-Indazol-5-yl)-benzooxazol-6-ol
[0127] 1H-Indazole-5-carbonitrile hydrochloride (1.5 g) was
suspended in ethanol (15 mL) at 0.degree. C. The mixture was
saturated with hydrogen chloride while the temperature was raised
to room temperature. The reaction was then left overnight.
Diethylether was added and the resulting precipitate,
1H-indazole-5-carboximidic acid ethyl ester.2HCl (1.39 g), was
collected and dried in under high vacuum. According to synthetic
method B except that pyridine (540 ?L) was added, from
4-amino-benzene-1,3-diol hydrochloride (0.36 mg) and
1H-indazole-5-carboximidic acid ethyl ester hydrochloride (0.71 g)
was obtained the title compound (0.34 mg) as an off-white solid.
The work-up of the reaction was modified as follows: After
completion of the reaction, the mixture was diluted with ether and
water, and filtered. The solids were washed with water and ether,
and dried under high vacuum. MS: 252.4 (MH.sup.+); HPLC t.sub.R:
1.80 min.
Example 36
5-(7-Bromo-5-methoxy-benzooxazol-2-yl)-pyrrolidin-2-one
[0128] 2-Amino-6-bromo-4-methoxy-phenol (0.40 g),
1,3-dimethylaminopropyl)-3-ethylcarbodiimide (1.06 g),
1-hydroxybenzotriazole (0.50 g), dimethylaminopyridine (0.22 g) and
DL-5-oxo-pyrrolidine-2-carboxylic acid (0.25 g) were reacted
together in methylene chloride (7.3 mL). After 3 h, the reaction
was diluted with methylene chloride (10 mL) and washed successively
with 1N HCl (10 mL), sat. aq. NaHCO.sub.3 (10 mL), sat. aq. NaCl
(10 mL). The organic layer was dried over sodium sulfate, filtered
through celite and concentrated. Chromatography on silica gel
(eluant: 0 to 20% methanol:methylene chloride) afforded
5-oxo-pyrrolidine-2-carboxylic acid
(3-bromo-2-hydroxy-5-methoxy-phenyl)-amide (0.18 g). MS: 370.2
(100%), 372.2 (90%) (MH.sup.+); HPLC t.sub.R: 1.51 min.
5-Oxo-pyrrolidine-2-carboxylic acid
(3-bromo-2-hydroxy-5-methoxy-phenyl)-amide was cyclized according
to synthetic method R to obtain the title compound (0.09 g) after
purification on silica gel (eluant: 0 to 30% methanol:methylene
chloride). MS: 352.2 (100%), 354.2 (95%) (MH.sup.++CH.sub.3CN),
311.2 (20%), 313.2 (20%) (MH.sup.+).; HPLC t.sub.R: 1.84 min.
Reference Example 37
5-Methoxy-2-aminobenzenethiol hydrochloride
[0129] Prepared following literature procedure: Can. J. Chem.
43,1965, 2610.
Example 38
6-Hydroxy-2-R-benzothiazol
[0130] ##STR47##
[0131] 5-Methoxy-2-aminobenzenethiol hydrochloride was dissolved in
1-methyl-2-pyrrolidinone. After triethylamine (1 eq) was added, the
mixture was stirred at room temperature for 10 min. Acid chloride
(1 eq) was then added and the mixture was heated at 100.degree. C.
for 1 h. The mixture was then cooled to room temperature and 1N
NaOH was added to PH 9. The solid was collected by filtration and
washed with water. The solid was further dried on vacuum to give
6-methoxy-2-R-benzothiazol (yield step 1) which was treated with
boron tribromide under standard procedure to give
6-hydroxy-2-R-benzothiazol (yield step 2). TABLE-US-00006 R group
##STR48## ##STR49## ##STR50## ##STR51## yield on step 1 68 29 70 85
yield on step 2 66 68 39 85 Mass Spec MH.sup.+ 234 218 253 297
Example 39
4-Methyl-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
[0132] ##STR52## To a solution of
4-bromo-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (1.0 g, 2.86
mmol) in dry toluene (25 mL) was added potassium carbonate (4.2 g,
30.4 mmol), methylboronic acid (0.92 g, 15.3 mmol) and
tetrakis(triphenylphosphine)palladium (0.44 g, 0.38 mmol). The
mixture was heated to 100.degree. C. for 24 h, then cooled to room
temperature. The mixture was diluted with ethylacetate and washed
with water, saturated sodium carbonate and brine. After
chromatographic purification gave
4-methyl-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole as a white
solid (0.63 g, 77% yield). Mass spec: MH.sup.+=286.
Example 40
4-Methyl-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0133] ##STR53##
4-Methyl-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (174 mg, 0.61
mmol) was treated with boron tribromide under standard condition to
give 4-methyl-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole (105 mg,
67% yield) as a yellow solid. Mass spec: MH.sup.+=258.
Example 41
4-Bromomethyl-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
[0134] ##STR54##
4-Methyl-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (0.53 g, 1.87
mmol), N-bromosuccinimide (0.33 g, 1.87mmol) and benzoyl peroxide
(9 mg) were suspended in carbon tetrachloride (6 mL) and refluxed
for 3.5 h, then cooled to room temperature. Solvent was evaporated
and the mixture was purified by chromatography to give
4-bromomethyl-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (0.47 g,
70% yield) as a white solid. Mass spec: MH.sup.+=364.
Example 42
4-Cyanomethyl-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
[0135] ##STR55## To a solution of
4-bromomethyl-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole ( 0.47
g, 1.29 mmol) in ethanol (6 mL) was added a solution of potassium
cyanide (0.1 g, 1.54 mmol) in water (0.5 mL). The mixture was
refluxed for 1.5 h, then cooled to room temperature. Ethanol was
evaporated and the mixture was extracted with ethylacetate. After
chromatographic purification gave
4-cyanomethyl-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole as a
yellow solid (0.29 g, 73% yield). Mass spec: MH.sup.+=311.
Example 43
4-Cyanomethyl-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0136] ##STR56##
4-Methylcyano-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (0.29 g,
0.94 mmol) was treated with boron tribromide under standard
condition to give
4cyanomethyl-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole (116 mg,
44% yield) as a yellow solid. Mass spec: MH.sup.+=283.
Example 44
4-Trimethylsilylacetylene-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
[0137] ##STR57##
4-Bromo-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (0.3 g, 0.857
mmol), (trimethylsilyl)acetylene (0.48 mL, 3.43 mmol),
triethylamine (0.48 mL, 3.43 mmol) and
tetrakis(triphenylphosphine)palladium (0.2 g, 0.171 mmol) were
suspended in THF (6 mL) in a sealed tube and heated to 70.degree.
C. for 24 h, then cooled to room temperature. Ethylacetate and
water were added, the ethylacetate layer was washed with brine.
After chromatigraphic purification gave
4-trimethylsilylacetylene-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
(0.132 g, 42% yield) as a yellow solid. Mass spec:
MH.sup.+=368.
Example 45
4-Acetylene-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
[0138] ##STR58## To a solution of
4-trimethylsilylacetylene-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
(0.216 g, 0.588 mmol) in THF (5 mL) was added 1N sodium hydroxide
(1.18 mL, 1.18 mmol) and the solution was stirred at room
temperature for 3 h. THF was then evaporated. Water and
ethylacetate were added. Combined ethylacetate were concentrated.
After chromatigraphic purification gave
4-acetylene-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (0.113 g,
65% yield) as a yellow solid. Mass spec: MH.sup.+=296
Example 46
4-Acetylene-6 hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0139] ##STR59##
4-Acetylene-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (93 mg,
0.315 mmol) was treated with boron tribromide under standard
condition to give
4-acetylene-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole (47 mg,
46% yield) as a yellow solid. Mass spec: MH.sup.+=268
Example 47
4-Methylcarboxy-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
[0140] ##STR60##
4-Bromo-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (1.0 g, 2.86
mmol), triethylamine (0.96 mL, 7.15 mmol) palladium acetate (31 mg,
0.143 mmol) and 1,3-bis(diphenylphosphino)propane (57 mg, 0.143
mmol) were suspended in methanol (7 mL) and DMSO (7 mL). The
mixture was heated to 75.degree. C. and bubbled with CO for 20 min.
The mixture was then heated under CO for 48 h. After cooling to
room temperature, brine was added. The mixture was extracted with
ethylacetate. Combined ethylacetate were concentrated. After
chromatographic purification gave
4-methylcarboxy-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (0.37
g, 39% yield) as a white solid. Mass spec: MH.sup.+=330.
Example 48
4-Carboxy-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
[0141] ##STR61## To a solution of
4-methylcarboxy-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (0.37
g, 1.12 mmol) in THF (6 mL) and water (3 mL) was added 1N sodium
hydroxide (2.24 mL, 2.24 mmol) and the solution was stirred at room
temperature for 24 h. THF was then evaporated. 1 N HCl was added to
PH 1. The solid was collected by filtration and washed with water
to give 4-carboxy-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
(0.29 g, 82% yield) as a white solid. Mass spec: MH.sup.+=316.
Example 49
4-Carboxy-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0142] ##STR62##
4-Carboxy-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (50 mg,
0.159 mmol) was treated with boron tribromide under standard
condition to give
4-carboxy-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole (34 mg, 74%
yield) as a yellow solid. Mass spec: MH.sup.+=288. 1H NMR
(DMSO-d6): 10.24 (s, 1H), 10.16 (s, 1H), 7.91 (d, 2H), 7.67 (s,
1H), 7.48 (s, 1H), 6.95 (d, 2H).
Example 50
4-Carboxyamide-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0143] ##STR63##
[0144] 4-Carboxy-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (1
eq), 1-hydroxybenzotriazole (2.4 eq), dimethylamine or methylamine
(3.7 eq) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
hydrochloride (2.3 eq) were suspended in DMF and stirred at room
temperature for 5 min. Triethylamine (4.1 eq) was added and the
mixture was stirred at room temperature for 24 h. [when
R.sub.1.dbd.R.sub.2.dbd.H, only
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.05
eq) and 1-hydroxybenzotriazole ammonia (2.46 eq) were added].
Saturated sodium bicarbonate was added and the mixture was extrated
with ethylacetate. Combined ethylacetate were washed with brine,
concentrated. After chromatographic purification gave the product
(yield step 1) which was further treated with boron tribromide
under standard condition to give
4-carboxyamide-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole (yield
step 2). TABLE-US-00007 R.sub.1/R.sub.2 R.sub.1.dbd.R.sub.2.dbd.Me
R.sub.1.dbd.H, R.sub.2.dbd.Me R.sub.1.dbd.R.sub.2.dbd.H yield on
step 1 86 67 91 yield on step 2 100 70 44 Mass Spec MH.sup.+ 315
301 287
Example 51
6-Methoxy-2-(2-bromo-3-methoxy-phenyl)-benzothiazole
[0145] ##STR64## 5-Methoxy-2-aminobenzenethiol hydrochloride (83 g,
4.33 mmol) was dissolved in 1-methyl-2-pyrrolidinone (10 mL). After
triethylamine (0.60 mL, 4.33 mmol) was added, the mixture was
stirred at room temperature for 10 min. 2-Bromo-3-methoxybenzoyl
chloride (1.08 g, 4.33 mmol) in 1-methyl-2-pyrrolidinone (10 mL)
was then added and the mixture was heated at 100.degree. C. for 40
min. The mixture was cooled to room temperature and 1N NaOH was
added to PH 9. The mixture was extracted with ethylacetate and
combined ethylacetate were washed with brine, concentrated. After
chromatographic purification gave
6-methoxy-2-(2-bromo-3-methoxy-phenyl)-benzothiazole (0.98 g, 65%
yield) as a yellow solid. Mass spec: MH.sup.+=350.
Example 52
6-Hydroxy-2-(2-bromo-3-hydroxy-phenyl)-benzothiazole
[0146] ##STR65##
6-Methoxy-2-(2-bromo-3-methoxy-phenyl)-benzothiazole (0.144 g,
0.411 mmol) was treated with boron tribromide under standard
condition to give
6-hydroxy-2-(2-bromo-3-hydroxy-phenyl)-benzothiazole (35 mg, 26%
yield) as a yellow solid. Mass spec: MH.sup.+=322.
Example 53
6-Methoxy-2-(2-methyl-3-methoxy-phenyl)-benzothiazole
[0147] ##STR66## To a solution of
6-methoxy-2-(2-bromo-3-methoxy-phenyl)-benzothiazole (0.128, 0.366
mmol) in dry toluene (4 mL) was added potassium carbonate (0.404 g,
2.92 mmol), methylboronic acid (88 mg, 1.47 mmol) and
tetrakis(triphenylphosphine)palladium (42 mg, 0.036 mmol). The
mixture was heated to 100.degree. C. for 3 h, then cooled to room
temperature. The mixture was diluted with ethylacetate and washed
with water, saturated sodium carbonate and brine. After
chromatographic purification gave
6-methoxy-2-(2-methyl-3-methoxy-phenyl)-benzothiazole (66 mg, 63%
yield) as a white solid. Mass spec: MH.sup.+=286.
Example 54
6-Hydroxy-2-(2-methyl-3-hydroxy-phenyl)-benzothiazole
[0148] ##STR67##
6-Methoxy-2-(2-methyl-3-methoxy-phenyl)-benzothiazole (64 mg, 0.224
mmol) was treated with boron tribromide under standard condition to
give 6-hydroxy-2-(2-methyl-3-hydroxy-phenyl)-benzothiazole (52 mg,
90% yield) as a yellow solid. Mass spec: MH.sup.+=258.
Example 55
6-Methoxy-2-(3-methoxy-phenyl)-benzothiazole
[0149] ##STR68##
[0150] To a solution of
6-methoxy-2-(2-bromo-3-methoxy-phenyl)-benzothiazole (0.182, 0.52
mmol) in dry DMF (3 mL) was added cesium carbonate (0.51 g, 1.57
mmol), triethylborane (0.58 mL, 1M in THF, 0.58 mmol) and
1,1'-bis(diphenylphosphino)ferrocene palladium dichloride
dichloromethane (18 mg, 0.022 mmol). The mixture was heated to
50.degree. C. for 24 h, then cooled to room temperature. Saturated
sodium bicarbonate was added and the mixture was extracted with
ethylacetate. Combined ethylacetate were washed with brine,
concentrated. After chromatographic purification gave
6-methoxy-2-(3-methoxy-phenyl)-benzothiazole (54 mg, 38% yield) as
a white solid. Mass spec: MH.sup.+=272.
Example 56
6-Hydroxy-2-(3-hydroxy-phenyl)-benzothiazole
[0151] ##STR69## 6-Methoxy-2-(3-methoxy-phenyl)-benzothiazole (54
mg, 0.199 mmol) was treated with boron tribromide under standard
condition to give 6-hydroxy-2-(3-hydroxy-phenyl)-benzothiazole (15
mg, 31% yield) as a yellow solid. Mass spec: MH.sup.+=244.
Example 57
6-Hydroxy-2-(2-R-3-hydroxy-phenyl)-benzothiazole
[0152] ##STR70##
[0153] 5-Methoxy-2-aminobenzenethiol hydrochloride (1 eq) was
dissolved in 1-methyl-2-pyrrolidinone. After triethylamine (1 eq)
was added, the mixture was stirred at room temperature for 10 min.
2-R-3-methoxybenzoyl chloride (1 eq) in 1-methyl-2-pyrrolidinone
was then added and the mixture was heated at 100.degree. C.
(reaction time in the table). The mixture was cooled to room
temperature and 1N NaOH was added to PH 9. The mixture was
extracted with ethylacetate and combined ethylacetate were washed
with brine, concentrated. After chromatographic purification gave
6-methoxy-2-(2-R-3-methoxy-phenyl)-benzothiazole (yield step 1)
which was treated with boron tribromide under standard condition to
give the title compound (yield step 2). TABLE-US-00008 R group
--CH.sub.2CH.sub.3 --CH(CH.sub.3).sub.2 ##STR71## Reaction time on
step 1 45 min 300 min 30 min Yield on step 1 49 25 15 Yield on step
2 77 100 43 Mass spec MH.sup.+ 272 286 284
Example 58
4-Cyano-6-methoxy-2-(2-ethyl-3-methoxy-phenyl)-benzothiazole
[0154] ##STR72## The procedure was the same with
4-cyano-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole except using
2-ethyl-3-methoxy-benzoyl chloride instead of p-anisoyl chloride.
Mass spec: MH.sup.+=325.
Example 59
4-Cyano-6-hydroxy-2-(2-ethyl-3-hydroxy-phenyl)-benzothiazole
[0155] ##STR73##
4-Cyano-6-methoxy-2-(2-ethyl-3-methoxy-phenyl)-benzothiazole (0.1
g, 0.31 mmol) was treated with boron tribromide under standard
condition to give the title compound (30 mg, 33% yield) as a yellow
solid. Mass spec: MH.sup.+=297.
Example 60
4-Bromo-6-methoxy-2-(2-isopropyl-3-methoxy-phenyl)-benzothiazole
[0156] ##STR74## The procedure was the same with
4-bromo-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole except using
2-isopropyl-3-methoxy-benzoyl chloride instead of p-anisoyl
chloride. Mass spec: MH.sup.+=392.
Example 61
4-Bromo-6-hydroxy-2-(2-isopropyl-3-hydroxy-phenyl)-benzothiazol
[0157] ##STR75##
4-Bromo-6-methoxy-2-(2-isopropyl-3-methoxy-phenyl)-benzothiazole
(60 mg, 0.153 mmol) was treated with boron tribromide under
standard condition to give the title compound (30 mg, 54% yield) as
a yellow solid. Mass spec: MH.sup.+=364. ##STR76##
Example 62
4-Cyano-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0158] 4-Cyano-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole (0.18
g, 0.61 mmol) was suspended in boron tribromide (1M in methylene
chloride, 5.0 mL) and stirred at room temp under nitrogen for 18.0
hr. Reaction was poured into aqueous hydrochloric acid (1M). Solid
was collected by filtration and washed with water. Washed solid was
further purified by flash chromatography on silica affording the
title compound (150 mg, 92%) as a yellow solid. MH.sup.+=269;
.sup.1H NMR (300 MHZ, DMSO-d.sup.6,) d 10.42(s, 1H), 10.27(s, 1H),
7.91(d, J=8.7 hz, 2H), 7.77(d, J=2.4 hz, 1H), 7.36(d, J=2.4 hz,
1H), 6.94 (d, J=8.4 hz, 2H). The starting
4-Cyano-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole was prepared
as follows: ##STR77## a.
N-(2-Bromo-4-Methoxy-phenyl)-4-methoxy-benzamide To a solution
containing 2-bromo-4-methoxy-aniline.sup.1. (3.2 g, 15.8 mmol) in
pyridine (25 mL) was added p-anisoyl chloride (2.82 g, 16.5 mmol)
dropwise under nitrogen. The reaction was stirred at room temp for
1.0 hr. Reaction was poured cautiously into aqueous 1M hydrochloric
acid and extracted with ethyl acetate. Ethyl acetate extracts were
washed with: 1) hydrochloric acid (1.0M), 2) saturated brine and
concentrated in vacuo. This tan solid was suspended in a solution
containing: methanol (10 mL), methylene chloride (30 mL), and ethyl
acetate (5 mL), mixed for 5 min, then allowed to sit for 15 min,
yielding a white solid. This solid was collected by filtration and
dried under vacuum yielding the title compound (1.97 g, 37%) as a
white solid. Mass spec: MH.sup.+=336. .sup.1.Prepared following
literature procedure: Tet. (56) 2000, 1469 ##STR78## b.
N-(2-Bromo-4-Methoxy-phenyl)-4-methoxy-thiobenzamide
N-(2-Bromo-4-Methoxy-phenyl)-4-methoxy-benzamide (1.87 g, 5.6 mmol)
and Lawesson's reagent (1.35 g, 3.3 mmol) were suspended in
chlorobenzene (15 mL) and heated to reflux under nitrogen for 3.0
hr. Reaction was cooled, solvent removed under vacuum yielding a
yellow-orange solid. Solid was dissolved in ethyl acetate and
washed with: 1) 1N HCl, 2) saturated brine. Remove solvent under
vacuum. Solid was washed with hexane and dried under vacuum
yielding the title compound (1.93 g, 98%) as an orange solid. Mass
spec: MH.sup.+=352. ##STR79## c.
4-Bromo-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole
N-(2-Bromo-4-Methoxy-phenyl)-4-methoxy-thiobenzamide (1.5 g, 4.25
mmol) was wetted with ethanol (5.0 mL). 30% Aqueous sodium
hydroxide (10M, 3.4 mL) was added and stirred for 5 min. Water (6.8
mL) was added to provide a final suspension of 10% aqueous sodium
hydroxide. Aliquots (1 mL) of this mixture were added at 1 min
intervals to a heated (85.degree. C.) stirred solution containing
potassium ferricyanide (5.6 g, 17 mmol) in water (50 mL). Reaction
was kept at 85.degree. C. for 30 min, and then cooled to room temp.
Cold water (120 mL) was added. Mixture was allowed to sit
undisturbed for 30 min. Precipitate was collected by filtration,
washed with water, and dried under vacuum. Solid was washed with
ether then dried under vacuum at 35.degree. C. yielding the title
compound (1.2 g, 80%) as a pale tan solid. Mass spec: MH.sup.+=350.
##STR80## d. 4-Iodo-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole
4-Bromo-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole (0.86 g, 2.46
mmol), copper (1) iodide (2.34 g, 12.3 mmol), and potassium iodide
(4.08 g, 24.6 mmol) were suspended in DMSO (12 mL) and heated to
175.degree. C. under nitrogen for 4 hr, then cooled to room temp.
Reaction was poured cautiously into aqueous hydrochloric acid
(1.0M), and extracted with ethyl acetate. Ethyl acetate extracts
were washed with: 1) hydrochloric acid (1.0M), 2) saturated sodium
thiosulfate, 3) saturated brine and concentrated in vacuo. Solid
was washed with methylene chloride/hexane (1:1), dried under vacuum
yielding the title compound (0.87 g, 89%) as a pale orange solid
MH.sup.+=398. ##STR81## e.
4-Cyano-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole
4-Iodo-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole (258 mg, 0.65
mmol) and copper (1) cyanide (87 mg, 0.975 mmol) were suspended in
DMF (6.0 mL) under nitrogen and heated to 150.degree. C. for 2.0 hr
then cooled to room temp. Reaction was poured cautiously into
aqueous 1M hydrochloric acid and extracted with ethyl acetate.
Ethyl acetate extracts were washed with: 1) hydrochloric acid
(1.0M), 2) saturated brine and concentrated in vacuo. Solid was
washed with ether/hexane (1:2), then dried under vacuum yielding
the title compound (0.185 g, 96%) as a pale tan solid. MH.sup.+=297
##STR82##
Example 63
4-Bromo-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0159] 4-Bromo-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole (200
mg, 0.57 mmol) was suspended in boron tribromide (1M in methylene
chloride, 7.5 mL) and stirred at room temp under nitrogen for 3.0
hr. Reaction was poured into aqueous hydrochloric acid (1M) and
extracted with ethyl acetate. Ethyl acetate extracts were washed
with: 1) hydrochloric acid (1.0M), 2) saturated brine and
concentrated in vacuo. Washed solid was further purified by flash
chromatography on silica affording the title compound (184 mg,
100%) as a yellow solid. MH.sup.+=322 ##STR83##
Example 64
4-Iodo-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0160] 4-Iodo-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole (175 mg,
0.44 mmol) [Compound of Example 1d] was suspended in boron
tribromide (1M in methylene chloride, 3.5 mL) and stirred at room
temp under nitrogen for 72.0 hr. Reaction was poured into aqueous
hydrochloric acid (1M) and extracted with ethyl acetate. Ethyl
acetate extracts were washed with: 1) hydrochloric acid (1.0M), 2)
saturated brine and concentrated in vacuo. Washed solid was further
purified by flash chromatography on silica affording the title
compound (158 mg, 97%) as a yellow solid. MH.sup.+=370
##STR84##
Example 65
4-Chloro-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0161] 4-Chloro-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole (0.27
g, 0.88 mmol) was suspended in boron tribromide (1M in methylene
chloride, 7.0 mL) and stirred at room temp under nitrogen for 18.0
hr. Reaction was poured cautiously into aqueous 1M hydrochloric
acid and extracted with ethyl acetate. Ethyl acetate extracts were
washed with: 1) hydrochloric acid (1.0M), 2) saturated brine and
concentrated in vacuo. Solid was washed with methylene
chloride/methanol (97:3, 10 mL), and dried under vacuum yielding
the title compound (0.240 g, 98%) as a tan solid. Mass spec:
MH.sup.+=278 The starting
4-Chloro-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole was prepared
as follows: ##STR85## a.
N-(2-Chloro-4-hydroxy-phenyl)-4-methoxy-benzamide To a solution
containing 2-chloro-4-hydoxy-aniline hydrochloride(1.44 g, 8 mmol)
in pyridine (10 mL) was added p-anisoyl chloride (1.38 g, 8.1 mmol)
dropwise under nitrogen. The reaction was stirred at room temp for
1.0 hr. Reaction was poured cautiously into aqueous 1M hydrochloric
acid and extracted with ethyl acetate. Ethyl acetate extracts were
washed with: 1) hydrochloric acid (1.0M), 2) saturated brine and
concentrated in vacuo. This solid was washed with ether and dried
under vacuum yielding the title compound (1.68 g, 76%) as a tan
solid. Mass spec: MH.sup.+=278. ##STR86## b.
N-(2-Chloro-4-methoxy-phenyl)-4-methoxy-benzamide
N-(2-Chloro-4-hydroxy-phenyl)-4-methoxy-benzamide (1.83 g, 6.59
mmol) and potassium carbonate (1.82 g, 13.2 mmol) were suspended in
DMF (15 mL). Methyl iodide (0.62 mL, 9.89 mmol) was added and
stirred at room temp under nitrogen for 15 min, then heated to
95.degree. C. for 18 hr. Reaction was cooled to room temp then
poured cautiously into aqueous 1M hydrochloric acid and extracted
with ethyl acetate. Ethyl acetate extracts were washed with: 1)
hydrochloric acid (1.0M), 2) saturated NaHCO.sub.3, 3) saturated
brine and concentrated in vacuo. Washed solid was further purified
by flash chromatography on silica affording the title compound
(1.31 g, 68%) as a white solid. MH.sup.+=292 ##STR87## c.
N-(2-Chloro-4-methoxy-phenyl)-4-methoxy-thiobenzamide
N-(2-Chloro-4-methoxy-phenyl)-4-methoxy-benzamide (0.62 g, 2.13
mmol) and Lawesson's reagent (0.52 g, 1.28 mmol) were suspended in
chlorobenzene (10 mL) and heated to reflux under nitrogen for 3.0
hr. Reaction was cooled, solvent removed under vacuum yielding a
yellow-orange solid which was further purified by flash
chromatography on silica affording the title compound (0.60 g, 92%)
as a yellow solid. Mass spec: MH.sup.+=308 ##STR88## d.
4-Chloro-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole
N-(2-Chloro-4-Methoxy-phenyl)-4-methoxy-thiobenzamide (0.307 g, 1
mmol) was wetted with ethanol (4.0 mL). 30% Aqueous sodium
hydroxide (10M, 0.8 mL) was added and stirred for 5 min. Water (2.4
mL) was added to provide a final suspension of 10% aqueous sodium
hydroxide. Aliquots (1 mL) of this mixture were added at 1 min
intervals to a heated (85.degree. C.) stirred solution containing
potassium ferricyanide (1.32 g, 4 mmol) in water (20 mL). Reaction
was kept at 85.degree. C. for 30 min, and then cooled to room temp.
Reaction was poured cautiously into aqueous 1M hydrochloric acid
and extracted with ethyl acetate. Ethyl acetate extracts were
washed with: 1) hydrochloric acid (1.0M), 2) saturated NaHCO.sub.3,
3) saturated brine and concentrated in vacuo. Solid was washed with
ether, dried under vacuum yielding the title compound (0.285 g,
93%) as a white solid. Mass spec: MH.sup.+=306 ##STR89##
Example 66
2-(4-Hydroxy-phenyl)-4-Trifluormethyl-6-hydroxy-benzothiazole
[0162] 6-Methoxy-2-(4-Methoxy-phenyl)
4-Trifluoromethyl--benzothiazole (0.08 g, 0.23 mmol) was suspended
in boron tribromide (1M in methylene chloride, 5.0 mL) and stirred
at room temp under nitrogen for 18.0 hr. Reaction was poured into
aqueous hydrochloric acid (1M). Solid was collected by filtration
and washed with water. Washed solid was further purified by flash
chromatography on silica affording the title compound (21 mg, 29%)
as a white solid. MH.sup.+=311 The starting
6-Methoxy-2-(4-Methoxy-phenyl) 4-Trifluoromethyl-benzothiazole was
prepared as follows: ##STR90## a. 6-Methoxy-2-(4-Methoxy-phenyl)
4-Trifluoromethyl-benzothiazole
4-Iodo-6-methoxy-2-(4-Methoxy-phenyl)-benzothiazole (397 mg, 1.0
mmol) [Compound of Example 1d] and copper (0) powder (150 mg, 2.36
mmol) were suspended in pyridine (15.0 mL) under nitrogen in a Parr
bomb equipped with gas inlet. Trifluoromethyl Iodide (6.0 g, 30.6
mmol)was added via the gas inlet. Reaction was heated to
165.degree. C. for 48.0 hr then cooled to room temp. Remove
pyridine under vacuum Reaction was suspended in ethyl acetate/1MHCl
(200 mL, 1: 1), and filtered. The ethyl acetate extract was washed
with: 1) hydrochloric acid (1.0M), 2) saturated NaHCO.sub.3 3)
saturated sodium thiosulfate, 4) saturated brine and concentrated
in vacuo. Residue was further purified by flash chromatography on
silica affording the title compound (0.16 g, 47%) as a tan solid.
Mass spec: MH.sup.+=339 ##STR91##
Example 67
2-(4-Hydroxy-phenyl)-6-hydroxy-benzothiazole
[0163] 6-Methoxy-2-(4-methoxy-phenyl)-benzothiazole (134 mg, 0.49
mmol) and pyridine hydrochloride ( 1.34 g, 11.6 mmol) were heated
to 200.degree. C. under nitrogen for 40 min, and then cooled to
room temp. Reaction was poured cautiously into aqueous hydrochloric
acid (1M) and extracted with ethyl acetate. Ethyl acetate extracts
were washed with: 1) hydrochloric acid (1.0M), 2) saturated brine
and concentrated in vacuo. Residue was washed with ether/hexane
(1:4), dried under vacuum yielding the title compound (119 mg,
100%) as a yellow solid. MH.sup.+=244 The starting
6-Methoxy-2-(4-methoxy-phenyl)-benzothiazole was prepared as
follows: ##STR92## a. 2-Bromo-6-methoxy-benzothiazole To a solution
containing dry Copper (11) bromide (2.68 g, 12 mmol), tri(ethylene
glycol) dimethyl ether (5 g) in acetonitrile (150 mL) was added
isoamyl nitrite (2 mL, 15 mmol). Reaction was stirred at room temp
under nitrogen for 30 min. To this suspension was added, dropwise,
a solution (obtained by sonification) containing
2-amino-6-methoxy-benzothiazole (1.8 g, 10 mmol) and tri(ethylene
glycol) dimethyl ether (5 g) in acetonitrile (50 mL). Reaction was
stirred at room temp for 10 min, and then heated to 50.degree. C.
for 3 hr. Reaction was cooled to room temp, poured cautiously into
aqueous 6M hydrochloric acid and extracted with ethyl acetate.
Ethyl acetate extracts were washed with: 1) hydrochloric acid
(1.0M), 2) saturated brine and concentrated in vacuo. Residue was
crystallized from ether/hexane (1:10) yielding the title compound
(1.48 g, 61%) MH.sup.+=244 Supernatant solution was concentrated,
dried under vacuum yielding 2,7-dibromo-6-methoxy-benzothiazole
(0.45 g, 14%) as a yellow solid. MH.sup.+=322 ##STR93## b.
6-Methoxy-2-(4-methoxy-phenyl)-benzothiazole
2-Bromo-6-methoxy-benzothiazole (244 mg, 1 mmol),
4-methoxy-phenyl-boronic acid (198 mg, 1.3 mmol),
tetrakis(triphenylphosphine) palladium (0) (58 mg, 0.05 mmol), and
cesium fluoride (380 mg, 2.5 mmol) were suspended in acetonitrile
(10 mL) and heated to reflux for 90 min under nitrogen. Reaction
was cooled to room temp, poured cautiously into aqueous 1M
hydrochloric acid and extracted with ethyl acetate. Ethyl acetate
extracts were washed with: 1) hydrochloric acid (1.0M), 2)
saturated NaHCO.sub.3, 3) saturated brine and concentrated in
vacuo. Washed solid was further purified by flash chromatography on
silica affording the title compound (271 mg, 100%) as a pale yellow
solid. MH.sup.+=272 ##STR94##
Example 68
7-Chloro-6-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0164] 7-Bromo-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole (65 mg,
0.186 mmol)) and pyridine hydrochloride (1.6 g, 13.8 mmol) were
heated to 200.degree. C. under nitrogen for 45 min, and then cooled
to room temp. Reaction was poured cautiously into aqueous 1M
hydrochloric acid and extracted with ethyl acetate. Ethyl acetate
extracts were washed with: 1) hydrochloric acid (1.0M), 2)
saturated brine and concentrated in vacuo. Residue was further
purified by flash chromatography on silica affording the title
compound (51 mg, 99%) as a white solid. MH.sup.+=278 The starting
7-Bromo-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole was prepared
as follows: ##STR95## a.
7-Bromo-6-methoxy-2-(4-methoxy-phenyl)-benzothiazole
2,7-Dibromo-6-methoxy-benzothiazole (94 mg, 0.29 mmol),
4-methoxy-phenyl-boronic acid (47 mg, 0.31 mmol),
tetrakis(triphenylphosphine) palladium (0) (19 mg, 0.015 mmol), and
cesium fluoride (110 mg, 0.725 mmol) were suspended in acetonitrile
(10 mL) and heated to reflux for 90 min under nitrogen. Reaction
was cooled to room temp, poured cautiously into aqueous 1M
hydrochloric acid and extracted with ethyl acetate. Ethyl acetate
extracts were washed with: 1) hydrochloric acid (1.0M), 2)
saturated NaHCO.sub.3, 3) saturated brine and concentrated in
vacuo. Washed solid was further purified by flash chromatography on
silica affording the title compound (71 mg, 70%) as a white solid.
MH.sup.+=350 ##STR96##
Example 69
7-Cyano-5-hydroxy-2-(4-hydroxy-phenyl)-benzothiazole
[0165] 5-Methoxy-2-(4-methoxy-phenyl)-benzothiazole-7-carbonitrile
(0.04 g, 0.135 mmol) was suspended in boron tribromide (1M in
methylene chloride, 5.0 mL) and stirred at room temp, under
nitrogen for 48 hr. Reaction was poured into aqueous hydrochloric
acid (1M) and extracted with ethyl acetate. Ethyl acetate extracts
were washed with: 1) hydrochloric acid (1.0M), 2) saturated
NaHCO.sub.3, 3) saturated brine and concentrated in vacuo. This
material was further purified by chromatography on silica yielding
the title compound (14 mg, 39%) as a tan solid. Mass spec:
MH.sup.+=296
5-Methoxy-2-(4-hydroxy-phenyl)-benzothiazole-7-carbonitrile (7 mg,
18%) was also obtained as a white solid from this
chromatography.
[0166] The starting
7-Cyano-5-Methoxy-2-(4-methoxy-phenyl)-benzothiazole was prepared
as follows: ##STR97## a. 3-Amino-5-methoxy-phenol 3,5
Dimethoxy-aniline (1.53 g, 10 mmol) and pyridine hydrochloride (6.9
g, 60 mmol) were heated to 190.degree. C. under nitrogen for 60
min, and then cooled to room temp. Reaction was poured cautiously
into saturated NaHCO.sub.3 and extracted with ethyl acetate. Ethyl
acetate extracts were washed with: 1) saturated NaHCO.sub.3, 2)
saturated brine and concentrated in vacuo. Residue was further
purified by flash chromatography on silica affording the title
compound (600 mg, 44%) as a tan oil. MH.sup.+=139 ##STR98## b.
N-(3-Hydroxy-5-Methoxy-phenyl)-4-methoxy-benzamide To a solution
containing 3-Amino-5-methoxy-phenol (0.59 g, 4.28 mmol) in pyridine
(5 mL) was added p-anisoyl chloride (0.77 g, 4.49 mmol) dropwise
under nitrogen. The reaction was stirred at room temp for 18 hr.
Reaction was cautiously poured into aqueous 1M hydrochloric acid
and extracted with ethyl acetate. Ethyl acetate extracts were
washed with: 1) hydrochloric acid (1.0M), 2) saturated brine and
concentrated in vacuo. This tan solid was further purified by
chromatography on silica yielding the title compound (0.97 g, 83%)
as a tan solid. Mass spec: MH.sup.+=274. ##STR99## c.
Trifluoro-methanesulfonic-acid 3-methoxy-5-(4-methoxy-benzoyl
amino)-phenyl ester To a chilled (0.degree. C.) solution containing
N-(3-Hydroxy-5-Methoxy-phenyl)-4-methoxy-benzamide (0.546 g, 2
mmol), diisopropylethyl amine (646 mg, 5 mmol) in methylene
chloride (15 mL) was added, dropwise, a solution containing triflic
anhydride (0.846 g, 3 mmol) in methylene chloride (6 mL) under
nitrogen. The reaction was stirred at 0.degree. C. for 10 min and
then allowed to warm to room temp for 18 hr. Reaction was poured
cautiously into saturated NaHCO.sub.3 and extracted with ethyl
acetate. Ethyl acetate extracts were washed with: 1) saturated
NaHCO.sub.3, 2) saturated brine and concentrated in vacuo. This
material was further purified by chromatography on silica yielding
the title compound (0.44 g, 54%) as a tan oil. Mass spec:
MH.sup.+=406. ##STR100## d.
N-(3-Cyano-5-methoxy-phenyl)-4-methoxy-benzamide
Trifluoro-methanesulfonic-acid 3-methoxy-5-(4-methoxy-benzoyl
amino)-phenyl ester (0.41 g, 1 mmol) and potassium cyanide (0.163
g, 2.5 mmol) were suspended in DMF (5 ml) and heated to 120.degree.
C. under nitrogen for 18 hr. Reaction was cooled to room temp,
poured cautiously into saturated NaHCO.sub.3 and extracted with
ethyl acetate. Ethyl acetate extracts were washed with: 1)
saturated NaHCO.sub.3, 2) hydrochloric acid (1M), 3) saturated
brine and concentrated in vacuo. This material was further purified
by chromatography on silica yielding the title compound (0.17 g,
60%) as a tan solid. Mass spec: MH.sup.+=283. ##STR101## e.
N-(3-Cyano-5-methoxy-phenyl)-4-methoxy-thiobenzamide
N-(3-Cyano-5-methoxy-phenyl)-4-methoxy-benzamide (80 mg, 0.28 mmol)
and Lawesson's reagent (69 mg, 0.17 mmol) were suspended in
chlorobenzene (5 mL) and heated to reflux under nitrogen for 3.0
hr. Reaction was cooled, solvent removed under vacuum. Solid was
dissolved in ethyl acetate and washed with: 1) hydrochloric acid
(1.0M), 2) saturated NaHCO.sub.3, 3) saturated brine and
concentrated in vacuo yielding the title compound (83 mg, 100%) as
a yellow-orange solid. Mass spec: MH.sup.+=298 ##STR102## f.
7-Cyano-5-Methoxy-2-(4-methoxy-phenyl)-benzothiazole
N-(3-Cyano-5-methoxy-phenyl)-4-methoxy-thiobenzamide (90 mg, 0.30
mmol) was wetted with ethanol (3.0 mL). 30% Aqueous sodium
hydroxide (10M, 2.4 mL) was added and stirred for 5 min. Water (4.8
mL) was added to provide a final suspension of 10% aqueous sodium
hydroxide. Aliquots (1 mL) of this mixture were added at 1 min
intervals to a heated (85.degree. C.) stirred solution containing
potassium ferricyanide (398 mg, 1.21 mmol) in water (6 mL).
Reaction was kept at 85.degree. C. for 30 min, and then cooled to
room temp. Cold water (120 mL) was added. Extract with ethyl
acetate. Ethyl acetate extracts were washed with: 1) hydrochloric
acid (1.0M), 2) saturated NaHCO.sub.3, 3) saturated brine and
concentrated in vacuo. This material was further purified by
chromatography on silica yielding the title compound (44 mg, 49%)
as a white solid. Mass spec: MH.sup.+=296.
Example 70
2-(4-Amino-phenyl)-6-hydroxy-benzothiazole
[0167] ##STR103## 4-(6-Methoxy-benzothiazol-2-yl)-phenylamine (27
mg, 0.105 mmol) was suspended in boron tribromide (1M in methylene
chloride, 3.0 mL) and stirred at room temp under nitrogen for 18.0
h. Reaction was poured into saturated NaHCO.sub.3 and extracted
with ethyl acetate. Ethyl acetate extracts were washed with: 1)
saturated NaHCO.sub.3, 2) saturated brine and concentrated in
vacuo. This material was further purified by chromatography on
silica yielding the title compound (25 mg, 100%) as a tan solid.
Mass spec: MH.sup.+=243 The starting
4-(6-Methoxy-benzothiazol-2-yl)-phenylamine was prepared as
follows: ##STR104## 2-Bromo-6-methoxy-benzothiazole (244 mg, 1
mmol) [Example 67],
4-(4,4,5,5-tetramethyl-1,3,2-dioxboralan-2-yl)-aniline (285 mg, 1.3
mmol), tetrakis(triphenylphosphine) palladium (0) (58 mg, 0.05
mmol), and cesium fluoride (380 mg, 2.5 mmol) were suspended in
acetonitrile (10 mL) and heated to reflux for 90 min under
nitrogen. Reaction was cooled to room temp, poured cautiously into
saturated NaHCO.sub.3 and extracted with ethyl acetate. Ethyl
acetate extracts were washed with: 1) saturated NaHCO.sub.3, 2)
saturated brine and concentrated in vacuo. Washed solid was further
purified by flash chromatography on silica affording the title
compound (190 mg, 74%) as a pale yellow solid. MH.sup.+=272
Example 71
6-Bromo-2-(4-hydroxy-phenyl)-benzothiazole
[0168] ##STR105## 6-Bromo-2-(4-methoxy-phenyl)-benzothiazole (120
mg, 0.375 mmol) was suspended in boron tribromide (1M in methylene
chloride, 7.0 mL) and stirred at room temp under nitrogen for 18.0
h. Reaction was poured into saturated brine and extracted with
ethyl acetate. Ethyl acetate extracts were washed with: 1)
hydrochloric acid (1.0M), 2) saturated brine and concentrated in
vacuo. Residue was washed with hexane, and dried under vacuum
yielding the title compound (115 mg, 100%) as a tan solid. Mass
spec: MH.sup.+=306 The starting
6-Bromo-2-(4-methoxy-phenyl)-benzothiazole was prepared as follows:
##STR106## a. N-(4-Bromo-phenyl)-4-methoxy-benzamide To a solution
containing 4-bromo-aniline (1.0 g, 7 mmol) in pyridine (7 mL) was
added p-anisoyl chloride (0.77 mL, 7.1 mmol) dropwise under
nitrogen. The reaction was stirred at room temp for 30 min.
Reaction was poured cautiously into saturated NaHCO.sub.3 and
extracted with ethyl acetate. Ethyl acetate extracts were washed
with: 1) saturated NaHCO.sub.3, 2) saturated brine and concentrated
in vacuo. This solid was washed with a solution containing: ether/
hexane (1:5, 10 mL), dried under vacuum, yielding the title
compound (1.97 g, 92%) as a white solid. Mass spec: MH.sup.+=306.
##STR107## b. N-(4-Bromo-phenyl)-4-methoxy-thiobenzamide
N-(4-Bromo-phenyl)-4-methoxy-benzamide (1.95 g, 6.37 mmol) and
Lawesson's reagent (1.55 g, 3.82 mmol) were suspended in
chlorobenzene (25 mL) and heated to reflux under nitrogen for 3.0
h. Reaction was cooled, solvent removed under vacuum. Solid was
dissolved in ethyl acetate and washed with: 1) hydrochloric acid
(1.0M), 2) saturated brine and concentrated in vacuo. Residue was
further purified by chromatography on silica yielding the title
compound (1.85 g, 90%) as a yellow-orange solid. Mass spec:
MH.sup.+=322 ##STR108## c.
6-Bromo-2-(4-methoxy-phenyl)-benzothiazole
N-(4-Bromo-phenyl)-4-methoxy-thiobenzamide (483 mg, 1.5 mmol) was
wetted with ethanol (4.0 mL). 30% Aqueous sodium hydroxide (10M,
1.2 mL) was added and stirred for 5 min. Water (2.4 mL) was added
to provide a final suspension of 10% aqueous sodium hydroxide.
Aliquots (1 mL) of this mixture were added at 1 min intervals to a
heated (85.degree. C.) stirred solution containing potassium
ferricyanide (1.98 g, 6 mmol) in water (25 mL). Reaction was kept
at 85.degree. C. for 30 min, and then cooled to room temp. Cold
water (200 mL) was added. Mixture was allowed to sit undisturbed
for 30 min. Precipitate was collected by filtration, washed with
water, and dried under vacuum. Solid was dried under vacuum at
37.degree. C. yielding the title compound (0.45, 93%) as a pale
yellow solid. Mass spec: MH.sup.+=320.
Sequence CWU 1
1
1 1 70 DNA artificial sequence Synthetic sequence comprising a
Vitellogenin-gene Estrogen Response Element 1 ctagtctcga gaggtcactg
tgacctagat ctaggtcact gtgacctaga tctaggtcac 60 tgtgacctac 70
* * * * *