U.S. patent application number 09/796500 was filed with the patent office on 2001-07-26 for heterocyclic topoisomerase poisons.
This patent application is currently assigned to Rutgers, The State University of New Jersey. Invention is credited to Kim, Jung Sun, LaVoie, Edmond J., Liu, Leroy Fong, Rangarajan, Meera.
Application Number | 20010009919 09/796500 |
Document ID | / |
Family ID | 21813388 |
Filed Date | 2001-07-26 |
United States Patent
Application |
20010009919 |
Kind Code |
A1 |
LaVoie, Edmond J. ; et
al. |
July 26, 2001 |
Heterocyclic topoisomerase poisons
Abstract
The invention provides a topoisomerase poison of formula I: 1
wherein R.sub.1-R.sub.8 have any of the meanings defined in the
specification, or a pharmaceutically acceptable salt thereof, as
well as pharmaceutical compositions comprising a compound of
formula I or a salt thereof, intermediates useful for preparing a
compound of formula I, and therapeutic methods comprising
administering a compound of formula I or a salt thereof.
Inventors: |
LaVoie, Edmond J.;
(Princeton Junction, NJ) ; Kim, Jung Sun; (Pusan,
KR) ; Rangarajan, Meera; (Highland Park, NJ) ;
Liu, Leroy Fong; (Bridgewater, NJ) |
Correspondence
Address: |
SCHWEGMAN, LUNDBERG, WOESSNER & KLUTH
P.O. BOX 2938
MINNEAPOLIS
MN
55402
US
|
Assignee: |
Rutgers, The State University of
New Jersey
|
Family ID: |
21813388 |
Appl. No.: |
09/796500 |
Filed: |
February 28, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09796500 |
Feb 28, 2001 |
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09484402 |
Jan 14, 2000 |
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6221892 |
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09484402 |
Jan 14, 2000 |
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09023147 |
Feb 12, 1998 |
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6063801 |
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Current U.S.
Class: |
514/394 ;
548/305.4 |
Current CPC
Class: |
A61P 35/00 20180101;
C07D 235/26 20130101; A61P 31/10 20180101; C07D 235/30 20130101;
A61P 43/00 20180101; C07D 235/18 20130101; C07D 235/28 20130101;
C07D 491/04 20130101 |
Class at
Publication: |
514/394 ;
548/305.4 |
International
Class: |
A61K 031/4184; C07D
235/04 |
Goverment Interests
[0001] The invention described herein was made with government
support under grant CA-39662 awarded by the National Cancer
Institute. The United States Government has certain rights in the
invention.
Claims
1. A compound of formula I: 7wherein R.sub.1 and R.sub.2 are each
independently hydrogen, (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.6)cycloalk- yl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alko- xy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio,
(C.sub.1-C.sub.6)alkanoyloxy, aryl, heteroaryl,
aryl(C.sub.1-C.sub.6)alkyl, or heteroaryl(C.sub.1-C.sub.6)alk- yl;
or R.sub.1 and R.sub.2 taken together are methylenedioxy; or
R.sub.1 and R.sub.2 taken together are benzo; R.sub.3, R.sub.4, and
R.sub.5 are each independently selected from the group consisting
of hydrogen, (C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.2-C.sub.6)alkanoyloxy, aryl, heteroaryl,
aryl(C.sub.1-C.sub.6)alky- l, and heteroaryl(C.sub.1-C.sub.6)alkyl;
R.sub.6 and R.sub.7 are each independently hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalk- yl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)ycloalkyl(C.sub.1-C.sub.6)alkox- y, hydroxy, halo,
nitro, cyano, mercapto, carboxy, hydroxy(C.sub.1-C.sub.6- )alkyl,
halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, or (C.sub.1-C.sub.6)alkanoyloxy;
R.sub.8 is hydroxy, halo, nitro, cyano, mercapto, carboxy,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkoxy,
--NR.sub.aR.sub.b, halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, (C.sub.1-C.sub.6)alkanoyloxy, aryloxy,
or heteroaryloxy; or R.sub.8 is (C.sub.1-C.sub.6)alkyl substituted
by 1, 2, or 3 substituents independently selected from the group
consisting of hydroxy, nitro, cyano, mercapto, carboxy,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkoxy,
--NR.sub.aR.sub.b, trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio,
(C.sub.1-C.sub.6)alkanoyloxy, aryloxy, and heteroaryloxy; and each
R.sub.a and R.sub.b is independently hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbonyl, aryl, heteroaryl,
aryl(C.sub.1-C.sub.6)alkyl, heteroaryl(C.sub.1-C.sub.6)alkyl,
arylcarbonyl, or heteroarylcarbonyl; or R.sub.a and R.sub.b
together with the nitrogen to which they are attached are
pyrrolidino, piperidino, or morpholino. wherein any aryl,
heteroaryl, or benzo of R.sub.1-R.sub.5, R.sub.8, R.sub.a, and
R.sub.b may optionally be substituted by 1, 2, or 3 substituents
independently selected from the group consisting of
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alko- xy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio, and
(C.sub.1-C.sub.6)alkanoyloxy; or a pharmaceutically acceptable salt
thereof.
2. The compound of claim 1 wherein R.sub.2 is phenyl, optionally
substituted by 1, 2, or 3 substituents independently selected from
the group consisting of (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, hydroxy, halo, nitro, cyano, mercapto,
carboxy, halo(C.sub.1-C.sub.6)alky- l, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycar- bonyl,
(C.sub.1-C.sub.6)alkylthio, and (C.sub.1-C.sub.6)alkanoyloxy.
3. The compound of claim 1 wherein R.sub.2 is phenyl.
4. The compound of claim 1 wherein R.sub.1 is hydrogen.
5. The compound of claim 1 wherein R.sub.1 is halo.
6. The compound of claim 1 wherein R.sub.1 and R.sub.2 are each
halo.
7. The compound of claim 1 wherein R.sub.1 and R.sub.2 are each
bromo.
8. The compound of claim 1 wherein R.sub.3, R.sub.4, and R.sub.5
are each hydrogen.
9. The compound of claim 1 wherein R.sub.6 and R.sub.7 are each
hydrogen.
10. The compound of claim 1 wherein R.sub.6 is
(C.sub.1-C.sub.6)alkoxy, hydroxy, halo, nitro, cyano, mercapto,
carboxy, hydroxy(C.sub.1-C.sub.6)a- lkyl,
halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, or (C.sub.1-C.sub.6)alkanoyloxy.
11. The compound of claim 1 wherein R.sub.7 is
(C.sub.1-C.sub.6)alkoxy, hydroxy, halo, nitro, cyano, mercapto,
carboxy, hydroxy(C.sub.1-C.sub.6)a- lkyl,
halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, or (C.sub.1-C.sub.6)alkanoyloxy.
12. The compound of claim 1 wherein R.sub.8 is hydroxy, halo,
nitro, cyano, mercapto, carboxy, (C.sub.1-C.sub.6)alkoxy,
--NR.sub.aR.sub.b, halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, or (C.sub.1-C.sub.6)alkanoyloxy.
13. The compound of claim 1 wherein R.sub.8 is
(C.sub.1-C.sub.6)alkyl substituted by 1, 2, or 3 hydroxy, nitro,
cyano, mercapto, carboxy, (C.sub.1-C.sub.6)alkoxy,
--NR.sub.aR.sub.b, trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbonyl, (C.sub.1-C.sub.6)alkylthio, or
(C.sub.1-C.sub.6)alkanoyloxy.
14. The compound of claim 1 wherein R.sub.8 is
(C.sub.1-C.sub.6)alkyl substituted by hydroxy, nitro, cyano,
mercapto, carboxy, (C.sub.1-C.sub.6)alkoxy, --NR.sub.aR.sub.b,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbonyl, (C.sub.1-C.sub.6)alkylthio, or
(C.sub.1-C.sub.6)alkanoyloxy.
15. The compound of claim I wherein R.sub.8 is halo or
trifluoromethyl.
16. The compound of claim 1 wherein R.sub.8 is trifluoromethyl.
17. The compound
5-phenyl-2-[2'-(2"-mercaptobenzimidazol-5"-yl)benzimidazo-
l-5'-yl]benzimidazole;
5-phenyl-2-[2'(2"-trifluoromethylbenzimidazol-5"-yl- )
benzimidazol-5'-yl]benzimidazole; or
5,6-dibromo-2-[2'-(2"-trifluorometh-
yl-benzimidazol-5"-yl)benzimidazol-5'-yl]benzimidazole; or a
pharmaceutically acceptable salt thereof.
18. A pharmaceutical composition comprising a compound of claim 1,
in combination with a pharmaceutically acceptable carrier.
19. A therapeutic method comprising treating cancer by
administering to a mammal in need of such therapy, an effective
amount of a compound of claim 1.
20. A method comprising inhibiting cancer cells by contacting said
cancer cells with an effective amount of a compound of claim 1.
21. A compound of formula I: 8wherein R.sub.1 and R.sub.2 are each
independently (C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alko- xy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio,
(C.sub.1-C.sub.6)alkanoyloxy, aryl, heteroaryl,
aryl(C.sub.1-C.sub.6)alkyl, or heteroaryl(C.sub.1-C.sub.6)alk- yl;
or R.sub.1 and R.sub.2 taken together are methylenedioxy; R.sub.3,
R.sub.4, and R.sub.8 are each independently selected from the group
consisting of hydrogen, (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.6)cycloalk- yl, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbonyl, (C.sub.2-C.sub.6)alkanoyloxy,
aryl, heteroaryl, aryl(C.sub.1-C.sub.6)alky- l, and
heteroaryl(C.sub.1-C.sub.6)alkyl; R.sub.6 and R.sub.7 are each
independently hydrogen, (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.6)cycloalk- yl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alko- xy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio, or
(C.sub.1-.sub.6)alkanoyloxy; and R.sub.8 is hydrogen,
(C.sub.1-C.sub.6)alkyl, aryl, or heteroaryl; wherein any aryl or
heteroaryl of R.sub.1-R.sub.5 and R.sub.8 may optionally be
substituted by 1, 2, or 3 substituents independently selected from
the group consisting of (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alko- xy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio, and
(C.sub.1-C.sub.6)alkanoyloxy; or a pharmaceutically acceptable salt
thereof.
22. The compound of claim 21 wherein R.sub.1 and R.sub.2 are each
independently (C.sub.1-C.sub.6)alkoxy, hydroxy, halo, nitro, cyano,
mercapto, carboxy, hydroxy(C.sub.1-C.sub.6)alkyl,
halo(C.sub.1-C.sub.6)al- kyl, trifluoromethoxy, or aryl.
23. The compound of claim 21 wherein R.sub.1 and R.sub.2 taken
together are methylenedioxy.
24. The compound of claim 21 wherein R.sub.1 and R.sub.2 are each
halo.
25. The compound of claim 21 wherein R.sub.1 and R.sub.2 are each
bromo.
26. The compound of claim 21 wherein R.sub.1 and R.sub.2 are each
independently phenyl or methoxy.
27. The compound of claim 21 wherein R.sub.3, R.sub.4, and R.sub.5
are each hydrogen.
28. The compound of claim 21 wherein R.sub.6 is
(C.sub.1-C.sub.6)alkoxy, hydroxy, halo, nitro, cyano, mercapto,
carboxy, hydroxy(C.sub.1-C.sub.6)a- lkyl,
halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, or (C.sub.1-C.sub.6)alkanoyloxy.
29. The compound of claim 21 wherein R.sub.7 is
(C.sub.1-C.sub.6)alkoxy, hydroxy, halo, nitro, cyano, mercapto,
carboxy, hydroxy(C.sub.1-C.sub.6)a- lkyl,
halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, or (C.sub.1-C.sub.6)alkanoyloxy.
30. The compound of claim 21 wherein R.sub.8 is hydrogen.
31. The compound of claim 21 wherein R.sub.8 is
(C.sub.1-C.sub.6)alkyl.
32. The compound of claim 21 wherein R.sub.8is aryl, optionally
substituted by 1, 2, or 3 substituents independently selected from
the group consisting of (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloal- kyl(C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alk- oxy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio, and
(C.sub.1-C.sub.6)alkanoyloxy.
33. The compound of claim 21 wherein R.sub.8 is heteroaryl,
optionally substituted by 1, 2, or 3 substituents independently
selected from the group consisting of (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alko- xy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio, and
(C.sub.1-C.sub.6)alkanoyloxy.
34. The compound
5,6-dibromo-2-[2'-(benzimidazol-5"-yl)benzimidazol-5'yl]
benzimidazole, or a pharmaceutically acceptable salt thereof.
35. A pharmaceutical composition comprising a compound of claim 21,
in combination with a pharmaceutically acceptable carrier.
36. A therapeutic method comprising treating cancer by
administering to a mammal in need of such therapy, an effective
amount of a compound of claim 21.
37. A method comprising inhibiting cancer cells by contacting said
cancer cells with an effective amount of a compound of claim
21.
38. A therapeutic method comprising treating fungal infection by
administering to a mammal afflicted with a fungal infection, an
effective antifungal amount of a compound of claim 1 or claim 21.
Description
BACKGROUND OF THE INVENTION
[0002] DNA-topoisomerases are enzymes present in the nuclei of
cells where they catalyze the breaking and rejoining of DNA
strands, controlling the topological state of DNA. Recent studies
also suggest that topoisomerases are involved in regulating
template supercoiling during RNA transcription. There are two major
classes of mammalian topoisomerases. DNA-topoisomerase-I catalyzes
changes in the topological state of duplex DNA by performing
transient single-strand breakage-union cycles. In contrast,
mammalian topoisomerase II alters the topology of DNA by causing a
transient enzyme bridged double-strand break, followed by strand
passing and resealing. Mammalian topoisomerase II has been further
classified as Type II .alpha. and Type II .beta.. The antitumor
activity associated with agents which are topoisomerase poisons is
associated with their ability to stabilize the enzyme-DNA cleavable
complex. This drug-induced stabilization of the enzyme-DNA
cleavable complex effectively converts the enzyme into a cellular
poison.
[0003] Several antitumor agents in clinical use have potent
activity as mammalian topoisomerase II poisons. These include
adriamycin, actinomycin D, daunomycin, VP-16, and VM-26 (teniposide
or epipodophyllotoxin).
[0004] In contrast to the number of clinical and experimental drugs
which act as topoisomerase II poisons, there are currently only a
limited number of agents which have been identified as
topoisomerase I poisons. Camptothecin and its structurally-related
analogs are among the most extensively studied topoisomerase I
poisons. Recently, bi- and terbenzimidazoles (Chen et al., Cancer
Res. 1993, 53, 1332-1335; Sun et al., J. Med. Chem. 1995, 38,
3638-3644; Kim et al., J Med. Chem. 1996, 39, 992-998), certain
benzo[c]phenanthridine and protoberberine alkaloids and their
synthetic analogs (Makhey et al., Med. Chem. Res. 1995, 5, 1-12;
Janin et al., J. Med. Chem 1975, 18, 708-713; Makhey et al.,
Bioorg. & Med. Chem. 1996, 4, 781-791), as well as the fungal
metabolites, bulgarein (Fujii et al., J. Biol. Chem. 1993, 268,
13160-13165) and saintopin (Yamashita et al., Biochemistry 1991,
30, 5838-5845) and indolocarbazoles (Yamashita et al., Biochemistry
1992, 31, 12069-12075) have been identified as topoisomerase I
poisons.
[0005] Presently, a need exists for novel anti-cancer agents, for
anti-cancer agents that exhibit improved activity, and for
anti-cancer agents that exhibit fewer side-effects or improved
selectivity compared to existing agents.
SUMMARY OF THE INVENTION
[0006] The present invention provides compounds that exhibit
inhibitory activity against topoisomerase I, and compounds that are
effective cytotoxic agents against cancer cells, including
drug-resistant cancer cells. Accordingly there is provided a
compound of the invention which is a compound of formula I: 2
[0007] wherein
[0008] R.sub.1 and R.sub.2 are each independently hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6) alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkoxy, hydroxy, halo,
nitro, cyano, mercapto, carboxy, hydroxy(C.sub.1-C.sub.6)alkyl,
halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, (C.sub.1-C.sub.6)alkanoyloxy, aryl,
heteroaryl, aryl(C.sub.1-C.sub.6)alky- l, or
heteroaryl(C.sub.1-C.sub.6)alkyl; or R.sub.1 and R.sub.2 taken
together are methylenedioxy; or R.sub.1 and R.sub.2 taken together
are benzo;
[0009] R.sub.3, R.sub.4, and R.sub.5 are each independently
selected from the group consisting of hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.2-C.sub.6)alkanoyloxy, aryl, heteroaryl,
aryl(C.sub.1-C.sub.6)alkyl, and heteroaryl(C.sub.1-C.sub.6)al-
kyl;
[0010] R.sub.6 and R.sub.7 are each independently hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alko- xy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)akyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio, or
(C.sub.1-C.sub.6)alkanoyloxy;
[0011] R.sub.8 is hydroxy, halo, nitro, cyano, mercapto, carboxy,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alko- xy,
--NR.sub.aR.sub.b, halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, (C.sub.1-C.sub.6)alkanoyloxy, aryloxy,
or heteroaryloxy; or R.sub.8 is (C.sub.1-C.sub.6)alkyl substituted
by 1, 2, or 3 substituents independently selected from the group
consisting of hydroxy, nitro, cyano, mercapto, carboxy,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkoxy,
--NR.sub.aR.sub.b, trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio,
(C.sub.1-C.sub.6)alkanoyloxy, aryloxy, and heteroaryloxy; and
[0012] each of R.sub.a and R.sub.b is independently hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbonyi, aryl, heteroaryl,
aryl(C.sub.1-C.sub.6)alkyl, heteroaryl(C.sub.1-C.sub.6)alkyl,
arylcarbonyl, or heteroarylcarbonyl; or R.sub.a and R.sub.b
together with the nitrogen to which they are attached are
pyrrolidino, piperidino, or morpholino.
[0013] wherein any aryl, heteroaryl, or benzo of R.sub.1-R.sub.5,
R.sub.8, R.sub.a, and R.sub.b may optionally be substituted by 1,
2, or 3 substituents independently selected from the group
consisting of (C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alko- xy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio, and
(C.sub.1-C.sub.6)alkanoyloxy;
[0014] or a pharmaceutically acceptable salt thereof
[0015] The invention also provides a compound of the invention
which is a compound of formula I: 3
[0016] wherein
[0017] R.sub.1 and R.sub.2 are each independently
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)- alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.- 6)alkoxy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio,
(C.sub.1-C.sub.6)alkanoyloxy, aryl, heteroaryl,
aryl(C.sub.1-C.sub.6)alkyl, or heteroaryl(C.sub.1-C.sub.6)alk- yl;
or R.sub.1 and R.sub.2 taken together are methylenedioxy;
[0018] R.sub.3, R.sub.4, and R.sub.5 are each independently
selected from the group consisting of hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.2-C.sub.6)alkanoyloxy, aryl, heteroaryl,
aryl(C.sub.1-C.sub.6)alkyl, and heteroaryl(C.sub.1-C.sub.6)al-
kyl;
[0019] R.sub.6 and R.sub.7 are each independently hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)alko- xy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio, or
(C.sub.1-C.sub.6)alkanoyloxy; and
[0020] R.sub.8 is hydrogen, (C.sub.1-C.sub.6)alkyl, aryl, or
heteroaryl;
[0021] wherein any aryl or heteroaryl of R.sub.1-R.sub.5 and
R.sub.8 may optionally be substituted by 1, 2, or 3 substituents
independently selected from the group consisting of
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.6)- alkyl,
(C.sub.1-C.sub.6)alkoxy,
(C.sub.3-C.sub.6)cycloalkyl(C.sub.1-C.sub.- 6)alkoxy, hydroxy,
halo, nitro, cyano, mercapto, carboxy,
hydroxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl,
trifluoromethoxy, (C.sub.1-C.sub.6)alkanoyl,
(C.sub.1-C.sub.6)alkoxycarbo- nyl, (C.sub.1-C.sub.6)alkylthio, and
(C.sub.1-C.sub.6)alkanoyloxy;
[0022] or a pharmaceutically acceptable salt thereof
[0023] The invention also provides a pharmaceutical composition
comprising a compound of formula I, or a pharmaceutically
acceptable salt thereof, in combination with a pharmaceutically
acceptable carrier.
[0024] The invention also provides a therapeutic method comprising
inhibiting cancer cells by administering to a mammal (e.g. a human)
in need of such therapy, an amount of a compound of formula I, or a
pharmaceutically acceptable salt thereof, effective to inhibit said
cancer cells.
[0025] The invention also provides a method comprising inhibiting
cancer cells by contacting said cancer cells in vitro or in vivo
with an amount of a compound of formula I, or a pharmaceutically
acceptable salt thereof, effective to inhibit said cancer cells,
i.e. to inhibit their activity, such as their ability to divide,
migrate, or proliferate.
[0026] The invention also provides a compound of formula I for use
in medical therapy (preferably for use in treating cancer, e.g.
solid tumors; or for use as an antifungal agent), as well as the
use of a compound of formula I for the manufacture of a medicament
useful for the treatment of cancer, e.g. solid tumors, and the use
of a compound of formula I for the manufacture of a medicament
useful for the treatment of fungal infeaction.
[0027] The invention also provides processes and novel
intermediates disclosed herein which are useful for preparing
compounds of the invention. Some of the compounds of formula I are
useful to prepare other compounds of formula I.
BRIEF DESCRIPTION OF THE FIGURES
[0028] FIG. 1 shows the structure of representative compounds of
the invention and other compounds (1, 6, 9, 14, 17, and 19).
[0029] FIG. 2 shows biological data for representative compounds of
formula I.
DETAILED DESCRIPTION
[0030] The following definitions are used, unless otherwise
described: halo is fluoro, chloro, bromo, or iodo. Alkyl, alkoxy,
etc. denote both straight and branched groups; but reference to an
individual radical such as "propyl" embraces only the straight
chain radical, a branched chain isomer such as "isopropyl" being
specifically referred to. Aryl denotes a phenyl radical or an
ortho-fused bicyclic carbocyclic radical having about nine to ten
ring atoms in which at least one ring is aromatic. Heteroaryl
encompasses a radical attached via a ring carbon of a monocyclic
aromatic ring containing five or six ring atoms consisting of
carbon and one to four heteroatoms each selected from the group
consisting of non-peroxide oxygen, sulfur, and N(Y) wherein Y is
absent or is H, O, (C.sub.1-C.sub.4)alkyl, phenyl or benzyl, as
well as a radical of an ortho-fused bicyclic heterocycle of about
eight to ten ring atoms derived therefrom, particularly a
benzo-derivative or one derived by fusing a propylene,
trimethylene, or tetramethylene diradical thereto.
[0031] It will be appreciated by those skilled in the art that
compounds of the invention having a chiral center may exist in and
be isolated in optically active and racemic forms. Some compounds
may exhibit polymorphism. It is to be understood that the present
invention encompasses any racemic, optically-active, polymorphic,
or stereoisomeric form, or mixtures thereof, of a compound of the
invention, which possess the useful properties described herein, it
being well known in the art how to prepare optically active forms
(for example, by resolution of the racemic form by
recrystallization techniques, by synthesis from optically-active
starting materials, by chiral synthesis, or by chromatographic
separation using a chiral stationary phase) and how to determine
topoisomerase poisoning activity or cytotoxic activity using the
standard tests described herein, or using other similar tests which
are well known in the art.
[0032] Specific and preferred values listed below for radicals,
substituents, and ranges, are for illustration only; they do not
exclude other defined values or other values within defined ranges
for the radicals and substituents Specifically,
(C.sub.1-C.sub.6)alkyl can be methyl, ethyl, propyl, isopropyl,
butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, or hexyl;
(C.sub.3-C.sub.6)cycloalkyl can be cyclopropyl, cyclobutyl,
cyclopentyl, or cyclohexyl; (C.sub.3-C.sub.6)cycloalkyl(C.sub-
.1-C.sub.6)alkyl can be cyclopropylmethyl, cyclobutylmethyl,
cyclopentylmethyl, cyclohexylmethyl, 2-cyclopropylethyl,
2-cyclobutylethyl, 2-cyclopentylethyl, or 2-cyclohexylethyl;
(C.sub.1-C.sub.6)alkoxy can be methoxy, ethoxy, propoxy,
isopropoxy, butoxy, iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy, or
hexyloxy; (C.sub.1-C.sub.6)alkanoyl can be acetyl, propanoyl or
butanoyl; halo(C.sub.1-C.sub.6)alkyl can be iodomethyl,
bromomethyl, chloromethyl, fluoromethyl, trifluoromethyl,
trichloromethyl, 2-chloroethyl, 2-fluoroethyl,
2,2,2-trifluoroethyl, or pentafluoroethyl;
hydroxy(C.sub.1-C.sub.6)alkyl can be hydroxymethyl, 1-hydroxyethyl,
2-hydroxyethyl, 1-hydroxypropyl, 2-hydroxypropyl, 3-hydroxypropyl,
1-hydroxybutyl, 4-hydroxybutyl, 1-hydroxypentyl, 5-hydroxypentyl,
1-hydroxyhexyl, or 6-hydroxyhexyl; (C.sub.1-C.sub.6)alkoxycarbonyl
can be methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl,
isopropoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, or
hexyloxycarbonyl; (C.sub.1-C.sub.6)alkylthio can be methylthio,
ethylthio, propylthio, isopropylthio, butylthio, isobutylthio,
pentylthio, or hexylthio; (C.sub.1-C.sub.6)alkanoyloxy can be
formyloxy, acetoxy, propanoyloxy, butanoyloxy, isobutanoyloxy,
pentanoyloxy, or hexanoyloxy; aryl can be phenyl, indenyl, or
naphthyl; and heteroaryl can be furyl, imidazolyl, triazolyl,
triazinyl, oxazoyl, isoxazoyl, thiazolyl, isothiazoyl, pyrazolyl,
pyrrolyl, pyrazinyl, tetrazolyl, pyridyl, (or its N-oxide),
thienyl, pyrimidinyl (or its N-oxide), indolyl, isoquinolyl (or its
N-oxide) or quinolyl (or its N-oxide).
[0033] A specific value for R.sub.1 is hydrogen. Another specific
value for R.sub.1 is halo (e.g. bromo).
[0034] A specific value for R.sub.2 is phenyl, optionally
substituted by 1, 2, or 3 substituents independently selected from
the group consisting of (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, hydroxy, halo, nitro, cyano, mercapto,
carboxy, halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, and (C.sub.1-C.sub.6)alkanoyloxy.
Another specific value for R.sub.2 is phenyl.
[0035] A specific value for each of R.sub.3, R.sub.4, and R.sub.5
is hydrogen.
[0036] A specific value for R.sub.6 or R.sub.7 is
(C.sub.1-C.sub.6)alkoxy, hydroxy, halo, nitro, cyano, mercapto,
carboxy, hydroxy(C.sub.1-C.sub.6)a- lkyl,
halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, or (C.sub.1-C.sub.6)alkanoyloxy.
Another specific value for each of R.sub.6 and R.sub.7 is
hydrogen.
[0037] A specific value for R.sub.8 is hydroxy, halo, nitro, cyano,
mercapto, carboxy, (C.sub.1-C.sub.6)alkoxy, --NR.sub.aR.sub.b,
halo(C.sub.1-C.sub.6)alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, or (C.sub.1-C.sub.6)alkanoyloxy.
Another specific value for R.sub.8 is (C.sub.1-C.sub.6)alkyl
substituted by 1, 2, or 3 substituents independently selected from
the group consisting of hydroxy, nitro, cyano, mercapto, carboxy,
(C.sub.1-C.sub.6)alkoxy, --NR.sub.aR.sub.b, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbo- nyL
(C.sub.1-C.sub.6)alkylthio, and (C.sub.1-C.sub.6)alkanoyloxy.
Another specific value for R.sub.8 is (C.sub.1-C.sub.6)alkyl
substituted by hydroxy, nitro, cyano, mercapto, carboxy,
(C.sub.1-C.sub.6)alkoxy, --NR.sub.aR.sub.b, trifluoromethoxy,
(C.sub.1-C.sub.6)alkanoyl, (C.sub.1-C.sub.6)alkoxycarbonyl,
(C.sub.1-C.sub.6)alkylthio, or (C.sub.1-C.sub.6)alkanoyloxy.
[0038] A preferred value for R.sub.1 is hydrogen; for R.sub.2 is
phenyl; and for R.sub.8 is halo or trifluoromethyl. A more
preferred value for R.sub.8 is trifluoromethyl.
[0039] A preferred group of compounds of formula I wherein R.sub.1
and R.sub.2 are each halo (e.g. bromo).
[0040] Processes for preparing compounds of formula I are
illustrated by the following procedures in which the meanings of
the generic radicals are as given above unless otherwise
qualified.
[0041] A compound of formula I wherein R.sub.8 is hydroxy can be
prepared by treating an intermediate of formula II: 4
[0042] with urea, for example, using conditions similar to those
described in Example 1.
[0043] A compound of formula I wherein R.sub.8 is mercapto can be
prepared by treating an intermediate of formula II with
ethylxanthic acid potassium salt, for example, using conditions
similar to those described in Example 2.
[0044] A compound of formula I wherein R.sub.8 is amino can be
prepared by treating an intermediate of formula II with cyanogen
bromide, for example, using conditions similar to those described
in Example 3.
[0045] Compounds of formula I can generically be prepared by
reacting an intermediate diamine of formula III with an
intermediate aldehyde of formula IV. 5
[0046] The reaction can conveniently be carried out under
conditions similar to those described in the Examples (e.g.,
Example 4).
[0047] Compounds of formula I can also generally be prepared by
reacting an intermediate diamine of formula V with an aldehyde of
formula VI. 6
[0048] The reaction can conveniently be performed under conditions
similar to those described in the Examples (e.g. Example 12).
[0049] Compounds of formula I comprising a hydroxy group can be
prepared from a corresponding compound of formula I comprising a
methoxy group by treatment with boron tribromide, for example using
conditions similar to those described in Example 7.
[0050] Compounds of formula I comprising a amino group can be
prepared from a corresponding compound of formula I comprising a
acetamido group by hydrolysis using conditions similar to those
described in Example 8.
[0051] Intermediates of formula II, III, IV, V, and VI wherein
R.sub.1-R.sub.8 have any of the values, specific values, or
preferred values defined herein for the corresponding radicals in a
compound of formula I, are particularly useful for preparing
compounds of formula I.
[0052] In cases where compounds are sufficiently basic or acidic to
form stable nontoxic acid or base salts, administration of the
compounds as salts may be appropriate. Examples of pharmaceutically
acceptable salts are organic acid addition salts formed with acids
which form a physiological acceptable anion, for example, tosylate,
methanesulfonate, acetate, citrate, malonate, tartarate, succinate,
benzoate, ascorbate, .alpha.-ketoglutarate, and
.alpha.-glycerophosphate. Suitable inorganic salts may also be
formed, including hydrochloride, sulfate, nitrate, bicarbonate, and
carbonate salts.
[0053] Pharmaceutically acceptable salts may be obtained using
standard procedures well known in the art, for example by reacting
a sufficiently basic compound such as an amine with a suitable acid
affording a physiologically acceptable anion. Alkali metal (for
example, sodium, potassium or lithium) or alkaline earth metal (for
example, calcium) salts of carboxylic acids can also be made.
[0054] The compounds of formula I can be formulated as
pharmaceutical compositions and administered to a mammalian host,
such as a human patient in a variety of forms adapted to the chosen
route of administration, i.e., orally or parenterally, by
intravenous, intramuscular, topical or subcutaneous routes.
[0055] Thus, the present compounds may be systemically
administered, e.g., orally, in combination with a pharmaceutically
acceptable vehicle such as an inert diluent or an assimilable
edible carrier. They may be enclosed in hard or soft shell gelatin
capsules, may be compressed into tablets, or may be incorporated
directly with the food of the patient's diet. For oral therapeutic
administration, the active compound may be combined with one or
more excipients and used in the form of ingestible tablets, buccal
tablets, troches, capsules, elixirs, suspensions, syrups, wafers,
and the like. Such compositions and preparations should contain at
least 0.1% of active compound. The percentage of the compositions
and preparations may, of course, be varied and may conveniently be
between about 2 to about 60% of the weight of a given unit dosage
form. The amount of active compound in such therapeutically useful
compositions is such that an effective dosage level will be
obtained.
[0056] The tablets, troches, pills, capsules, and the like may also
contain the following: binders such as gum tragacanth, acacia, corn
starch or gelatin; excipients such as dicalcium phosphate; a
disintegrating agent such as corn starch, potato starch, alginic
acid and the like; a lubricant such as magnesium stearate; and a
sweetening agent such as sucrose, fructose, lactose or aspartame or
a flavoring agent such as peppermint, oil of wintergreen, or cherry
flavoring may be added. When the unit dosage form is a capsule, it
may contain, in addition to materials of the above type, a liquid
carrier, such as a vegetable oil or a polyethylene glycol. Various
other materials may be present as coatings or to otherwise modify
the physical form of the solid unit dosage form. For instance,
tablets, pills, or capsules may be coated with gelatin, wax,
shellac or sugar and the like. A syrup or elixir may contain the
active compound, sucrose or fructose as a sweetening agent, methyl
and propylparabens as preservatives, a dye and flavoring such as
cherry or orange flavor. Of course, any material used in preparing
any unit dosage form should be pharmaceutically acceptable and
substantially non-toxic in the amounts employed. In addition, the
active compound may be incorporated into sustained-release
preparations and devices.
[0057] The active compound may also be administered intravenously
or intraperitoneally by infusion or injection. Solutions of the
active compound or its salts can be prepared in water, optionally
mixed with a nontoxic surfactant. Dispersions can also be prepared
in glycerol, liquid polyethylene glycols, triacetin, and mixtures
thereof and in oils. Under ordinary conditions of storage and use,
these preparations contain a preservative to prevent the growth of
microorganisms.
[0058] The pharmaceutical dosage forms suitable for injection or
infusion can include sterile aqueous solutions or dispersions or
sterile powders comprising the active ingredient which are adapted
for the extemporaneous preparation of sterile injectable or
infusible solutions or dispersions, optionally encapsulated in
liposomes. In all cases, the ultimate dosage form should be
sterile, fluid and stable under the conditions of manufacture and
storage. The liquid carrier or vehicle can be a solvent or liquid
dispersion medium comprising, for example, water, ethanol, a polyol
(for example, glycerol, propylene glycol, liquid polyethylene
glycols, and the like), vegetable oils, nontoxic glyceryl esters,
and suitable mixtures thereof The proper fluidity can be
maintained, for example, by the formation of liposomes, by the
maintenance of the required particle size in the case of
dispersions or by the use of surfactants. The prevention of the
action of microorganisms can be brought about by various
antibacterial and antifungal agents, for example, parabens,
chlorobutanol, phenol, sorbic acid, thiomersal, and the like. In
many cases, it will be preferable to include isotonic agents, for
example, sugars, buffers or sodium chloride. Prolonged absorption
of the injectable compositions can be brought about by the use in
the compositions of agents delaying absorption, for example,
aluminum monostearate and gelatin.
[0059] Sterile injectable solutions are prepared by incorporating
the active compound in the required amount in the appropriate
solvent with various of the other ingredients enumerated above, as
required, followed by filter sterilization. In the case of sterile
powders for the preparation of sterile injectable solutions, the
preferred methods of preparation are vacuum drying and the freeze
drying techniques, which yield a powder of the active ingredient
plus any additional desired ingredient present in the previously
sterile-filtered solutions.
[0060] For topical administration, the present compounds may be
applied in pure form, i.e., when they are liquids. However, it will
generally be desirable to administer them to the skin as
compositions or formulations, in combination with a
dermatologically acceptable carrier, which may be a solid or a
liquid.
[0061] Useful solid carriers include finely divided solids such as
talc, clay, microcrystalline cellulose, silica, alumina and the
like. Useful liquid carriers include water, alcohols or glycols or
water-alcohol/glycol blends, in which the present compounds can be
dissolved or dispersed at effective levels, optionally with the aid
of non-toxic surfactants. Adjuvants such as fragrances and
additional antimicrobial agents can be added to optimize the
properties for a given use. The resultant liquid compositions can
be applied from absorbent pads, used to impregnate bandages and
other dressings, or sprayed onto the affected area using pump-type
or aerosol sprayers.
[0062] Thickeners such as synthetic polymers, fatty acids, fatty
acid salts and esters, fatty alcohols, modified celluloses or
modified mineral materials can also be employed with liquid
carriers to form spreadable pastes, gels, ointments, soaps, and the
like, for application directly to the skin of the user.
[0063] Examples of useful dermatological compositions which can be
used to deliver the compounds of formula I to the skin are known to
the art; for example, see Jacquet et al. (U.S. Pat. No. 4,608,392),
Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No.
4,559,157) and Wortzman (U.S. Pat. No. 4,820,508).
[0064] Useful dosages of the compounds of formula I can be
determined by comparing their in vitro activity, and in vivo
activity in animal models. Methods for the extrapolation of
effective dosages in mice, and other animals, to humans are known
to the art; for example, see U.S. Pat. No. 4,938,949.
[0065] Generally, the concentration of the compound(s) of formula I
in a liquid composition, such as a lotion, will be from about
0.1-25 wt-%, preferably from about 0.5-10 wt-%. The concentration
in a semi-solid or solid composition such as a gel or a powder will
be about 0.1-5 wt-%, preferably about 0.5-2.5 wt-%.
[0066] The amount of the compound, or an active salt or derivative
thereof, required for use in treatment will vary not only with the
particular salt selected but also with the route of administration,
the nature of the condition being treated and the age and condition
of the patient and will be ultimately at the discretion of the
attendant physician or clinician.
[0067] In general, however, a suitable dose will be in the range of
from about 0.5 to about 100 mg/kg, e.g., from about 10 to about 75
mg/kg of body weight per day, such as 3 to about 50 mg per kilogram
body weight of the recipient per day, preferably in the range of 6
to 90 mg/kg/day, most preferably in the range of 15 to 60
mg/kg/day.
[0068] The compound is conveniently administered in unit dosage
form; for example, containing 5 to 1000 mg, conveniently 10 to 750
mg, most conveniently, 50 to 500 mg of active ingredient per unit
dosage form.
[0069] Ideally, the active ingredient should be administered to
achieve peak plasma concentrations of the active compound of from
about 0.5 to about 75 .mu.M, preferably, about 1 to 50 .mu.M, most
preferably, about 2 to about 30 .mu.M. This may be achieved, for
example, by the intravenous injection of a 0.05 to 5% solution of
the active ingredient, optionally in saline, or orally administered
as a bolus containing about 1-100 mg of the active ingredient.
Desirable blood levels may be maintained by continuous infusion to
provide about 0.01-5.0 mg/kg/hr or by intermittent infusions
containing about 0.4-15 mg/kg of the active ingredient(s).
[0070] The desired dose may conveniently be presented in a single
dose or as divided doses administered at appropriate intervals, for
example, as two, three, four or more sub-doses per day. The
sub-dose itself may be further divided, e.g., into a number of
discrete loosely spaced administrations; such as multiple
inhalations from an insufflator or by application of a plurality of
drops into the eye.
[0071] The ability of a compound of the invention to effect
topoisomerase I mediated DNA cleavage can be determined using
pharmacological models that are well known to the art, for example,
using a model like Test A described below.
[0072] Test A. Topoisomerase I Cleavage Assay.
[0073] Representative compounds of the invention were evaluated in
a cleavage assay using recombinant topoisomerases I. This assay was
preformed as described by B. Gatto et al. Cancer Res., 1996, 56,
2795-2800. Human topoisomerase I was isolated as a recombinant
fusion protein using a T7 expression system. Plasmid YEpG was
purified by the alkali lysis method followed by phenol
deproteination and CsCl/ethidium isopycnic centrifiigation as
described by Maniatis, T.; Fritsch, E. F.; Sambrook, J. Molecular
Cloning, a Laboratory Manual; Cold Spring Harbor Laboratory: Cold
Spring Harbor, N.Y. 1982; pp 149-185. The end-labeling of the
plasmid was accomplished by digestion with a restriction enzyme
followed by end-filling with Klenow polymerase as previously
described by Liu, L. F.; Rowe, T. C.; Yang, L.; Tewey, K. M.; Chen,
G. L. "Cleavage of DNA by mammalian topoisomerase II," J. Biol.
Chem. 1983, 258, 15365. IC.sub.50 values were calculated after 4
days of continuous drug exposure. Topoisomerase I cleavage values
are reported as REC, Relative Effective Concentration (i.e.,
concentrations relative to compound 5, whose value is arbitrarily
assumed as 1) that is able to produce the same cleavage on the
plasmid DNA in the presence of human topoisomerase I.
[0074] The cytotoxic effects of a compound of the invention can be
determined using pharmacological models that are well known to the
art, for example, using a model like Test B described below.
[0075] Test B. Cytotoxicity Assay.
[0076] Cytotoxicity was determined using the MTT-microtiter plate
tetrazolinium cytotoxicity assay (MTA) (See Chen A. Y. et al.
Cancer Res. 1993, 53, 1332; Mosmann, T. J., J. Immunol. Methods
1983, 65, 55; and Carmichael, J. et al. Cancer Res. 1987, 47, 936).
The human lymphoblast RPMI 8402 and its camptothecin-resistant
variant cell line, CPT-K5 were provided by Dr. Toshiwo Andoh (Aichi
Cancer Center Research Institute, Nagoya, Japan) (see Andoh, T.;
Okada, K. "Drug resistance mechanisms of topoisomerase I drugs,"
Adv. in Pharmacology 1994, 29B, 93. The cytotoxicity assay was
performed using 96-well microtiter plates. Cells were grown in
suspension at 37.degree. C. in 5% CO.sub.2 and maintained by
regular passage in RPMI medium supplemented with 10% heat-
inactivated fetal bovine serum, L-glutamine (2 mM), penicillin (100
U/mL), and streptomycin (0.1 mg/mL). For determination of
IC.sub.50, cells were exposed continuously with varying
concentrations of drug and MTT assays were performed at the end of
the fourth day.
[0077] Data from Test A and Test B is shown in FIG. 2 for
representative compounds of the invention.
[0078] Compounds of formula I are potent topoisomerase I poisons.
Additionally, compounds of formula I exhibit cytotoxic activity
against RPMI 8402 cancer cells and camptothecin resistant CPT-K5
cells. Accordingly, compounds of formula I are useful as cytotoxic
agents, for the treatment of cancers, and in particular, solid
mammalian tumors or hematologic malignancies. Compounds of the
invention are also useful as pharmacological tools for in vitro and
in vivo study of topoisomerase function and activity.
[0079] As used herein, the term "solid mammalian tumors" includes
cancers of the head and neck, lung, mesothelioma, mediastinum,
esophagus, stomach, pancreas, hepatobiliary system, small
intestine, colon, rectum, anus, kidney, ureter, bladder, prostate,
urethra, penis, testis, gynecological organs, ovarian, breast,
endocrine system, skin central nervous system; sarcomas of the soft
tissue and bone; and melanoma of cutaneous and intraocular origin.
The term "hematological malignancies" includes childhood leukemia
and lymphomas, Hodgkin's disease, lymphomas of lymphocytic and
cutaneous origin, acute and chronic leukemia, plasma cell neoplasm
and cancers associated with AIDS. The preferred mammalian species
for treatment are humans and domesticated animals.
[0080] The compounds are also expected to exhibit some of the other
bioactivities observed for topoisomerase inhibitors, such as
antibacterial, antifungal, antiprotozoal, antielmintic and/or
antiviral activity. In particular, the compounds may exhibit
antifungal activity. Thus, the invention also provides a
therapeutic method to treat fungal infection comprising
administering to a mammal afflicted with a fungal infection, an
effective antifungal amount of a compound of the formula I, or a
pharmaceutically acceptable salt thereof.
[0081] The invention will now be illustrated by the following
non-limiting Examples, wherein unless otherwise stated: melting
points were determined with a Thomas-Hoover Unimelt capillary
melting point apparatus; column chromatography refers to flash
chromatography conducted on SiliTech 32-63 .mu.m, (ICN Biomedicals,
Eschwegge, Ger.) using the solvent systems indicated; infrared
spectral data (IR) were obtained on a Perkin-Elmer 1600 Fourier
transform spectrophotometer and are reported in cmil; proton
(.sup.1H NMR) and carbon (.sup.13C NMR) nuclear magnetic resonance
were recorded on a Varian Gemini-200 Fourier Transform
spectrometer; NMR spectra (200 MHZ .sup.1H and 50 MHZ .sup.13C)
were recorded in the deuterated solvent indicated with chemical
shifts reported in .delta. units downfield from tetramethylsilane
(TMS); coupling constants are reported in hertz (Hz); mass spectra
were obtained from Washington University Resource for Biomedical
and Bio-organic Mass Spectrometry within the Department of
Chemistry at Washington University, St. Louis, Mo.; and combustion
analyses were performed by Atlantic Microlabs, Inc., Norcross, Ga.,
and were within .+-.0.4% of the theoretical value.
EXAMPLES
Example 1
5-Phenyl-2-[2'(2"-hydroxybenzimidazol-5"-yl)benzimidazol-5'-yl]benzimidazo-
le (2).
[0082]
5-Phenyl-2-[2'-(3,4-diaminophenyl)benzimidazol-5'yl]benzimidazole,
(29.7 mg, 0.07 mmol) and urea (6 mg, 0.1 mmol) were dissolved in
DMF (0.4 ml). The mixture was refluxed at 150.degree. C. for 7
hours. The cooled reaction mixture when concentrated in vacuo and
washed with water provided 27.6 mg (88%) of pure yellow solid;
mp>280.degree. C.; IR (KBr) 3382, 3133, 1693, 1475, 1444, 1279;
.sup.1H NMR (DMSO-d.sub.6 +3 drops CF.sub.3COOH) .delta. 7.3
(d,1H,J=8.14), 7.46-7.80 (m,3H), 7.80 (d,2H,J=8.1),
7.89-8.01(m,4H), 8.05-8.14 (m,2H), 8.32 (d,1H,J=8.34), 8.63 (s,1H),
11.35 (s,1H); .sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH)
.delta. 108.07, 109.55, 111.78, 114.03, 114.57, 114.62, 115.3,
120.21, 122.53, 125.45, 125.82, 127.42, 128.1, 129.26, 130.81,
131.59, 132.66, 132.87, 135.15, 135.33, 139.20, 139.61, 149.32,
153.14, 155.71; HRMS (FAB) calculated for
C.sub.27H.sub.19N.sub.6(MH.sup.+) 443.1620 found 443.1625.
[0083] The intermediate
5-phenyl-2-[2'-(3,4-diaminophenyl)benzimidazol-5'y- l]benzimidazole
was prepared as follows.
[0084] a.
5-Phenyl-2-[2'-(3,4diaminophenyl)benzimidazol-5'-yl]benzimidazol-
e. A solution of
5-phenyl-2-[2'-(3,4-dinitrophenyl)benzimidazol-5'yl]benzi- midazole
(75 mg, 0.16 mmol) in ethyl acetate (50 mL) was reduced by
hydrogenation over 10% Pd/C (15 mg) for 90 minutes. The resulting
solution was passed through a bed of Celite and the ethyl acetate
was removed to give the diamine, which was used without further
purification.
[0085] The starting
5-phenyl-2-[2'-(3,4-dinitrophenyl)benzimidazol-5'yl]be- nzimidazole
can be prepared as described by J. S. Kim et al. J. Med. Chem.
1997, 40, 2818-2824.
Example 2
5-Phenyl-2-[2'-(2"-mercaptobenzimidazol-5"-yl)benzimidazol-5'-yl]benzimida-
zole (3).
[0086]
5-Phenyl-2-[2'-(3,4-diaminophenyl)benzimidazol-5'yl]benzimidazole,
(59.4 mg, 0.14 mmol) and ethylxanthic acid potassium salt (25.1 mg,
0.16 mmol) were refluxed in ethanol (1 ml) and distilled water (0.1
ml) overnight. The cooled reaction mixture was acidified to pH 3
with glacial acetic acid, volume condensed in vacuo and purified
directly by column chromatography. Elution with (40-100%) ethyl
acetate/n-hexanes provided 69% of yellow solid; mp>280.degree.
C.; IR (KBr) 3089, 2926, 2851, 1712, 1624, 1549, 1449, 1380, 1274,
1186, 1079; .sup.1H NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta.
7.33-7.42 (m,2H), 7.48-7.63 (t,4H), 7.71-7.82 (m,4H), 7.89 (s,1H),
8.04-8.06 (m,1H), 8.1-8.24 (m,1H), 8.42 (d,1H,J=2.4); .sup.13C NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 107.37, 107.15, 109.2,
109.68, 111.81, 111.73, 113.31, 113.53, 114.57, 114.64, 114.68,
115.56, 125.26, 127.44, 128.16, 129.33, 131.64, 132.93, 133.27,
139.1, 136.61, 139.62, 149.51, 152.94, 171.13; HRMS (FAB)
calculated for C.sub.27H.sub.19N.sub.6S (MH.sup.+) 459.1392, found
459.1403.
Example 3
5-Phenyl-2-[2'-(2"-aminobenzimidazol-5"-yl)benzimidazol-5'-yl]benzimidazol-
e (4).
[0087]
5-Phenyl-2-[2'-(3,4-diaminophenyl)benzimidazol-5'yl]benzimidazole,
(66 mg, 0.16 mmol) was dissolved in DMF (0.2 ml) and methanol (1
ml), and was added to cyanogen bromide (10% solution in water, 0.33
ml, 0.63 mmol). The reaction mixture was stirred overnight at room
temperature. The mixture was concentrated under reduced pressure.
Compound was recrystallized from methanol to give 47% of pale brown
solid; mp>280.degree. C.; IR (KBr) 3352, 3052, 2926, 1680, 1624,
1574, 1461, 1261,679; .sup.1H NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 7.46-7.62 (m,3H), 7.79 (d,2H,J=7.26),
7.87-7.96 (m,2H), 7.99-8.01 (m,2H), 8.20-8.26 (m,2H), 8.31 (s,1H),
8.72 (d,2H,J=9.12); .sup.13C NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 107.38, 110.89, 111.75, 111.78,
114.67,115.13, 115.24, 115.92, 123.59, 125.58, 126.84, 127.5,
127.53, 127.57, 127.76, 128.2, 129.43, 130.86, 131.87, 133.13,
138.74, 139.74, 150.10, 152.14, 153.42; HRMS (FAB) calculated for
C.sub.27H.sub.20N.sub.7 (MH.sup.+) 442.1780 found 442.1776.
Example 4
5-Phenyl-2-[2'(2"-trifluoromethylbenzimidazol-5"yl)benzimidazol-5'-yl]benz-
imidazole (5).
[0088] 5-Phenyl-2-[3,4-diaminophenyl]benzimidazole (0.160 g, 0.53
mmol) was heated with 5-formyl-2-trifluoromethylbenzimidazole (0.12
g, 0.54 mmol) in nitrobenzene (4 mL) at 145.degree. C. overnight.
Nitrobenzene was removed using a Kugelrohr and the compound
purified by column chromatography. (1-16%) methanol/ethyl acetate
provided 40% pure yellow compound; mp>280.degree. C.; IR (KBr)
3047, 2927, 1698, 1626, 1543, 1440, 1287, 1158; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH).delta. 7.44-7.59 (m,3H), 7.79
(d,2H,J=7.04), 7.87-8.01 (m,2H), 8.05-8.09 (m,2H), 8.15-8.19
(m,1H), 8.29-8.35 (m,2H), 8.70 (m,2H); .sup.13C NMR (DMSO-d.sub.6+3
drops CF.sub.3COOH) .delta. 111.79, 114.68, 114.92, 115.85, 117.53,
117.58, 118.37, 118.41, 119.58, 120.46, 124.14, 124.79, 125.71,
125.74, 127.47, 128.18, 129.36, 131.73, 132.99, 134.93, 137.44,
138.97, 139.02, 139.07, 139.66, 140.34, 149.81, 153.46; HRMS (FAB)
calculated for C.sub.28H.sub.18N.sub.6F.sub.3 (MH.sup.+) 495.1545
found 495.1543.
[0089] The intennediate 5-formyl-2-trifluoromethylbenzimidazole was
prepared as follows.
[0090] a. 3,4-Diaminobenzonitrile. 4-Amino-3-nitrobenzonitrile (3
g, 18.4 mmol) in ethyl acetate (120 ml) was reduced by
hydrogenation using 45 psi of H.sub.2 and 10% Pd-C (300 mg) for 1.5
hr. After passing through a bed of celite, the solvent was removed
in vacuo and 2.43 g (99%) of white solid was obtained; mp
144-145.degree. C.; IR (KBr) 3431, 2726, 2211, 1861, 1631, 1311,
1149, 722; .sup.1H NMR (CDCl.sub.3) .delta. 3.18-3.87 (brs,
4H),6.68 (d, 1H, J=8.06), 6.94 (d,1H,J=1.74), 7.02-7.27
(dd,1H,J=1.78, 8.06); .sup.13C NMR (CDCl.sub.3) .delta. 100.27,
115.86, 119.52, 122.19, 125.66, 135.94, 142.52; The crude diamine
obtained was typically used without further purification.
[0091] b. 5Cyano-2-trifluoromethylbenzimidazole. 3,4
Diaminobenzonitrile (200 mg, 1.5 mmol) was refluxed with
trifluoroacetic acid (0.3 ml) for 6 hours. The mixture was
neutralized with 2 N NaOH and extracted with ethyl acetate. The
ethyl acetate layer was dried over anhydrous Na.sub.2SO.sub.4.
Column with (0-10%) ethyl acetate/n-hexanes gave 88% of pure white
compound; mp 182-183.degree. C.; .sup.1H NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 7.72-7.77 (1H,dd,J=1.46,8.56), 7.88 (d,1H,
8.42), 8.35 (s,1H); .sup.13C NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 106.47, 106.64, 112.36, 117.31, 118.09,
119.44, 123.53, 123.81, 127.47; Anal. calculated for
C.sub.9H.sub.4N.sub.3F.sub.3: C; 51.20, H; 1.91, N; 19.90, found C;
51.45, H; 2.03, N; 19.68.
[0092] c. 5-Formyl-2-trifluoromethylbenzimidazole.
5-Cyano-2-trifluorometh- yl-benzimidazole (196 mg, 0.93 mmol) was
refluxed with HCOOH (14 ml), H.sub.2O (5 ml) and Ni-Al (0.9 g)
catalyst for 6 hours. The hot reaction mixture was filtered through
celite and concentrated in vacuo. The solution was basified with 2N
NaOH. Extraction with ethyl acetate followed by drying over
anhydrous sodium sulfate and concentration of the ethyl acetate
layer in vacuo gave the crude aldehyde. Purification was achieved
by column chromatography. Elution with 10/90 ethyl
acetate/n-hexanes gave 41% of pure white compound; mp
178-179.degree. C.; IR (KBr) 3210, 2737, 1699, 1552, 1328, 1187
(d), 986; .sup.1H NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta.
7.87-7.95 (m, 2H), 8.34(s,1H), 10.11 (s,1H)..sup.13C NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 106.63, 112.35, 116.72,
118.08, 121.87, 123.80, 124.04, 132.95, 192.65; HRMS (EI)
calculated for C.sub.9H.sub.5N.sub.2OF.sub.3 m/z 214.0354 found
214.0348.
[0093] The intermediate 4-amino-3-nitrobenzonitrile used in subpart
a above is commercially available (Aldrich Chemical Company)
[0094] The intermediate 5-phenyl-2-[3,4-diaminophenyl]benzimidazole
was prepared as follows.
[0095] d. 5-Phenyl-2-[3,4-diaminophenyl]benzimnidazole.
5-Phenyl-2-[3,4-nitrophenyl]benzimidazole (580 mg, 1.6 mmol) was
dissolved in 100 ml ethyl acetate and hydrogenation was carried out
using 10% Pd/C (100 mg) for 1.5 hours. The reaction mixture was
passed through a bed of celite and the bed washed with methanol.
The washings were concentrated in vacuo to yield (477 mg, 1.59
mmol) of the crude diamine in 98% yield. The diamine was not
characterized but used without purification; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 4.70 (brs,2H), 4.96
(brs, 2H), 6.62 (d,1H,8.02), 7.37 (m,8H), 7.53 (m,2H).
Example 5
5-Phenyl-2-[2'-(2"-hydroxymethylbenzimidazol-5"-yl]benzimidazol-5'-yl]benz-
imidazole (7).
[0096] 5-Phenyl-2-[3,4-diaminophenyl]benzimidazole (178 mg, 0.6
mmol) and 5-formyl-2-hydroxymethyl benzimidazole (104.2 mg, 0.6
mmol) were condensed in nitrobenzene (10 ml) overnight at
145.degree. C. Nitrobenzene was removed from the reaction mixture
with a Kugelrohr and the compound was purified directly by column
chromatography. Elution with (1-18%) methanol/ethyl acetate
provided 62% of yellow solid; mp>280.degree. C.; IR (Nujol)
3406, 2922, 2725, 1631, 1553, 1461, 1377; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 5.1 (s,2H), 7.45-7.59
(m,3H), 7.79 (d,2H,J=6.96), 8.23-8.28 (m,1H), 8.47 (d,1H, J=8.59),
8.68 (s,2H); .sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH)
.delta. 55.80, 111.69, 113.68, 114.59, 115.32, 115.77, 116.21,
118.31, 123.77, 124.52, 125.21, 125.61, 127.47, 127.49, 127.54,
128.14, 129.38, 131.64, 131.89, 132.91, 133.54, 137.66, 138.86,
139.68, 140.13, 150.25, 153.37, 157.62; HRMS (FAB) calculated for
C.sub.28H.sub.21N.sub.6O (MH.sup.+) 457.1777 found 457.1774.
[0097] The intermediate 5-formyl-2-hydroxymethyl benzimidazole was
prepared as follows.
[0098] a. 5-Cyano-2-hydroxymethylbenzimidazole. 3,4
Diaminobenzonitrile (0.2 g, 1.50 mmol) was heated with glycolic
acid (0.18 g, 2.37 mmol) in HCl (1.5 ml of 4N) for 2 hours. After
neutralization with 2M sodium carbonate the product was extracted
into ethyl acetate, dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuo to give a white colored solid. The compound
was purified by column chromatography. Elution with (75-100%) ethyl
acetate/n-hexanes gave 65.5% (133 mg, 0.76 mmol) of pure compound;
mp 172-173.degree. C.; IR (Nujol) 3350, 2928, 2219, 1621, 1536,
1302, 1217, 1036, 815; .sup.1H NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 4.88 (s,2H), 7.52-7.57 (dd,1H,J=1.46,8.38),
7.69 (d,1H,J=8.36), 7.95 (s,1H), .sup.13C NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 55.81, 107.99, 115.68, 118.64, 119.50,
129.00, 131.50, 134.39; HRMS (El) calculated for
C.sub.9H.sub.7N.sub.3O (m/z) 173.0590 found 173.0590.
[0099] b. 5Formyl-2-hydroxymethylbenzimidazole. Ni-Al (1.2 g) was
added to a solution of 5-cyano-2-hydroxymethylbenzimidazole (0.17
g, 0.97 mmol) in HCOOH (17 ml) and water (1.6 ml). The reaction
mixture was heated at 95.degree. C. for 5 hours. The hot mixture
was filtered through a bed of celite and the reaction flask and the
celite bed rinsed with water and then methanol. The combined
solution was concentrated in vacuo. After addition of water to this
residue, a white precipitate was formed. The pH of this suspension
was adjusted to 9 by dropwise addition of 2N NaOH. The product was
obtained by extraction with ethyl acetate. The ethyl acetate
extract was dried over anhydrous Na.sub.2SO.sub.4 and concentrated
in vacuo to give a pure white product (61% yield); mp
189-190.degree. C.; IR (KBr) 3309, 2924, 1674, 1619, 1434, 1291,
1073, 808; .sup.1H NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta.
5.07 (s,2H), 7.94 (d,1H,J=8.68), 8.04-8.09 (dd,1H,J=1.34,8.51),
8.32 (s,1H), 10.4 (s,1H); .sup.13C NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 55.76, 114.95, 116.81, 126.03, 131.45,
133.89, 135.11, 192.31; HRMS (EI) calculated for
C.sub.9H.sub.8N.sub.2O m/z 176.0586 found 176.0586.
Example 6
5-Phenyl-2-[2'-[2"-[2-(N-benzoyl)aminomethyl]benzimidazol-5"-yl]benzimidaz-
ol-5'-yl]benzimidazole (8).
[0100] 5-Phenyl-2-[3,4-diaminophenyl]benzimidazole (75 mg, 0.25
mmol) and 5-formyl-2-[(N-benzoyl)aminomethyl]benzimidazole (70 mg,
0.25 mmol,) were heated in nitrobenzene (6 ml) at 145.degree. C.
overnight. Nitrobenzene was removed with a Kugelrohr and the
compound was loaded on a column. (2-20%) methanol/ethyl acetate
gave 45% yellowish compound; mp>280.degree. C.; IR (KBr) 3204,
1637, 1542, 1442, 1384, 1292, 1026, 818; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 7.46-7.66 (m,6H), 7.80
(d, 2H, J=7.06), 7.87-8.1 (m, 7H), 8.21-8.25 (m,1H), 8.43-8.48
(m,1H), 8.67 (s,2H); .sup.13C NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 36.65, 107.37, 111.73, 114.53, 114.58,
115.26, 115.49, 116.01, 119.47, 122.49, 124.81, 125.56, 126.72,
127.41, 127.83, 128.04, 128.07, 128.54, 129.25, 131.56, 131.87,
132.12, 132.84, 133.17, 134.11, 136.21, 137.76, 139.08, 139.66,
149.67, 152.62, 155.64, 167.60; HRMS (FAB) calculated for
C.sub.35H.sub.26N.sub.7O (MH.sup.+) 560.2199 found 560.2209.
[0101] The intermediate
5-formyl-2-[(N-benzoyl)aminomethyl]benzimidazole was prepared as
follows.
[0102] a. 5-Cyano-2-[(N-benzoyl)aminomethyl]benzimidazole.
3,4-Diaminobenzonitrile (250 mg, 1.88 mmol) was finely ground in a
mortar with (0.34 g, 1.9 mmol) hippuric acid and intimately mixed
and then carefully fused for 3 hours. The temperature was then
raised to 160.degree. C. when water was evolved as bubbles. After
cooling the glassy mass was dissolved in ethyl acetate and purified
by column chromatography. (50-100%) ethyl acetate/n-hexanes yielded
86% of pale buff colored compound; mp 131-132.degree. C.; IR
(Nujol) 2826, 2221, 1635, 1313, 1019, 814; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 4.97 (d,2H,J=5.22),
7.52-7.61 (m,3H), 7.91-7.99 (m,4H), 8.37 (s,1H); .sup.13C NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 36.80, 107.73, 115.72,
118.81, 119.67127.83, 128.61, 128.64, 128.67, 128.78, 132.15,
132.20, 13322, 135.01, 156.13, 167.51; HRMS (EI) calculated for
C.sub.16H.sub.12N.sub.4O m/z 276.1011 found 276.1012.
[0103] b. 5-Formyl-2-[(N-benzoyl)aminomethyllbenzimidazole.
5-Cyano-2-[(N-benzoyl)aminomethyl]benzimidazole (0.33 g, 1.2 mmol)
was dissolved in formic acid (18 ml). Water (5 ml) and Ni-Al (1.14
g) were added to it. The mixture was heated under nitrogen at
95.degree. C. for 6 hours and while hot passed through a celite
bed. The bed was washed with methanol and the washings concentrated
in vacuo. The pH of the solution was adjusted with 2N NaOH to 9.0
and extracted with ethyl acetate. The extractions were dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was purified over column chromatography using (50-100%) ethyl
acetate/n-hexanes to give 26% of pure cream colored product; mp
108-109.degree. C.; IR (KBr) 3314, 2933, 1687, 1646, 1539, 1289,
806; .sup.1H NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 4.99
(d,1H,J=5.12), 7.54-7.63 (m,4H), 7.93-8.07 (m,3H), 8.34 (s,1H),
10.15 (s,1H); .sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH)
.delta. 36.83, 115.10, 117.18, 125.80, 127.85, 128.71, 132.26,
132.33, 133.2, 133.68, 155.77, 167.54, 192.54; HRMS (EI) calculated
for C.sub.16H.sub.13N.sub.3O- .sub.2 m/z 279.1008 found
279.1003.
Example 7
5-Phenyl-2-[2'-[2"-(2-hydroxyethyl)benzimidazol-5"-yl]benzimidazol-5'-yl]b-
enzimidazole (10)
[0104]
5-Phenyl-2-[2'-[2"-(2-methoxyethyl)benzimidazol-5"-yl]benzimidazol--
5'-yl]benzimidazole (30 mg, 0.06 mmol, Example 10) was suspended in
freshly distilled ethyl acetate (30 ml) to which was added 10
equivalence of BBr.sub.3 (0.62 ml, 1.0M) at -78.degree. C. After
stirring at room temperature overnight, the reaction was quenched
by adding water (10 ml). The mixture was basified to pH 9.0 with
ammonium hydroxide and extracted with ethyl acetate and dried over
anhydrous Na.sub.2SO.sub.4. The crude mixture was separated on a
chromatotron, (0-30%) methanol/ethyl acetate to give 10.6 mg of the
product in 33% yield as a brown solid. mp>280.degree. C.; IR
(KBr) 3401, 3150, 2876, 1632, 1540, 1442, 1388, 1268, 1013, 823;
.sup.1H NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 3.35
(t,2H), 3.98 (t,2H), 7.42-7.56 (m,3H), 7.76 (d,2H,J=7.7), 7.85-7.98
(m,2H), 8.06-8.19 (m,3H), 8.33 (d,1H, J=8.5), 8.44 (d,1H, J=8.8),
8.73 (s,2H); .sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH)
.delta. 30.75, 58.09, 111.71, 114.36, 114.53, 115.28, 115.98,
116.02, 118.28,119.43, 122.37, 124.71, 124.76, 125.43, 125.73,
127.40, 128.1, 129.25, 131.55, 131.64, 132.83, 133.95, 135.26,
137.79, 139.06, 139.64, 149.67, 152.63, 156.21; HRMS (FAB)
calculated for C.sub.29H.sub.23N.sub.6O (MH.sup.+) 471.1933 found
471.1935.
Example 8.
5-Phenyl-2-[2'-[2"-(2-aminoethyl)benzimidazol-5"-yl]benzimidazol-5'-yl]ben-
zimidazole (11)
[0105]
5-Phenyl-2-[2'-[2"-(2-acetamidoethyl)benzinidazol-5"-yl)]benzimidaz-
ol-5'-yl]benzimidazole (35 mg, 0.07 mmuyol, Example 9) was
hydrolyzed with 2N HCl (5 ml) at 110.degree. C. for 3 hours. The
cooled reaction mixture was basified with arnmonium hydroxide to pH
9.0 and purified by column chromatography. Elution with 50:50
methanol/ethyl acetate provided 75% yellow solid; mp>280.degree.
C.; IR (Nujol) 3375, 2719, 1553, 1461,1377; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 3.45-3.52 (m,4H),
7.45-7.56 (m,2H), 7.79(d, 2H, J=7.04), 7.88-8.00 (m,2H), 8.07-8.27
(m,5H),8.42 (d,1H, J=8.92), 8.68 (d,2H,J=4.76); .sup.13C NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 25.52, 36.36, 111.74,
113.77, 114.62, 115.33, 115.76, 116.17, 118.33, 123.75, 123.94,
124.75, 125.61, 127.48, 128.16, 129.39, 131.71, 132.98, 133.11,
134.77, 137.73, 138.84, 139.71, 140.12, 150.34, 153.43, 153.61;
HRMS (FAB) calculated for C.sub.27H.sub.19N.sub.6O (MH.sup.+)
470.2093 found 470.2089.
Example 9
5Phenyl-2-[2'-(2"(2-acetamidoethyl)benzimidazol-5"-yl)]benzimidazol-5'-yl]
benzimidazole (12).
[0106] 5-Phenyl-2-[3,4-diaminophenyl]benzimidazole (124 mg, 0.41
mmol) and 5-formyl-2-(2-acetamidoethyl)benzimidazole (95 mg, 0.41
mmol) were condensed in nitrobenzene (7 ml) overnight at
145.degree. C. Nitrobenzene was removed with the aid of a Kugelrohr
and the compound purified directly by flash column chromatography.
Elution with (1-15 %) methanol/ethyl acetate provided 69% of yellow
solid; mp>280.degree. C.; IR (KBr) 3064, 2954, 2862, 1654, 1556,
1442, 1286; .sup.1H NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta.
1.81 (s,3H), 3.18-3.35 (m,2H), 3.61-3.64 (m,2H), 7.45-7.6 (m,3H),
7.79 (d,2H,J=7.04 ), 7.87-8.00 (m,2H), 8.07-8.30 (m,4H), 8.46
(d,1H,J=8.5), 8.71 (s,2H); .sup.13C NMR (DMSO-d.sub.63 drops
CF.sub.3COOH) .delta. 22.59, 27.89, 36.41, 111.73, 113.66, 114.59,
115.22, 115.69, 116.17, 118.61, 124.04, 125.22, 125.66, 127.47,
127.56, 128.14, 128.18, 129.36, 131.62, 131.84, 132.89, 133.69,
137.10, 138.94, 139.54, 139.68, 150.16, 153.24, 155.40, 158.47,
170.27; HRMS (FAB) calculated for C.sub.31H.sub.25N.sub.7O
(MH.sup.+) 512.2199 found 512.2201.
[0107] The intermediate 5-formyl-2-(2-acetamidoethyl)benzimidazole
was prepared as follows.
[0108] a. 5-Cyano-2-(2-aminoethyl)benzimidazole. A solution of 3,4
diaminobenzonitrile (1 g, 7.52 mmol) and .beta.-alanine (1.0 g,
11.3 mmol) in HCl (8 ml, 6N) was refluxed for 24 hrs. At this time
the diamine was barely detectable by TLC. After neutralization with
2N NaOH, the compound was directly loaded on the column. Elution
with (75-100%) ethyl acetate/n-hexanes and further with (1-25%)
methanol/ethyl acetate gave 46% (0.65 g) of pure yellow compound;
mp 105-106.degree. C.; IR (KBr) 3438, 2855, 2734, 2212, 1625, 1569,
1483, 1457, 1385, 1222, 1153; .sup.1H NMR (CD.sub.3OD) .delta.
3.09-3.13 (m,4H), 7.49-7.66 (m,2H), 7.91 (d,1H,J=1.46); .sup.13C
NMR (CD.sub.3OD) .delta. 33.13, 40.99, 106.27, 116.57, 121.04,
121.19, 127.08, 140.40, 142.59, 158.82; HRMS (EI) calculated for
C.sub.10H.sub.10N.sub.4 m/z 186.0905 found 186.0906.
[0109] b. 5-Cyano-2-(2-acetamidoethyl)benzimidazole.
5-Cyano-2-(2-aminoethyl)benzimidazole (0.4 g, 2.15 mmol) was
refluxed in dry THF (10 ml), acetic anhydride (0.4 ml) and
triethylamine (0.4 ml) for 4 hrs. The mixture was neutralized with
2N NaOH and extracted with ethyl acetate to give the crude
acetamide. The acetamide was purified using flash column
chromatography. Elution with (0-15%) methanol/ethyl acetate gave
70% of pure white compound; mp 218-219.degree. C.; IR (KBr) 3233,
3053, 2225, 1653, 1573, 1442, 1374, 1303, 1058, 815; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 1.78 (s,3H), 3.26
(t,2H), 3.53-3.62 (2H, J=6.45, 12.38), 7.89-7.95 (dd, 1H, J=1.42,
8.48), 8.00 (d, J=8.22,1H), 8.19 (t,1H), 8.45 (d,1H,J=1.42);
.sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 22.57,
27.82, 36.71, 108.06, 115.52, 118.62, 119.33, 129.02, 131.29,
134.26, 156.23, 170.18; Anal. calculated for
C.sub.12H.sub.12N.sub.4O.1/4H2O: C; 61.92, H; 5.41, N; 24.07 found
C; 62.60, H; 5.41, N; 23.82.
[0110] c. 5-Formyl-2-(2-acetamidoethyl)benzimidazole.
5-Cyano-2-(2-acetamidoethyl)benzimidazole (94.6 mg, 0.41 mmol) was
refluxed in HCOOH (5.9 ml), water (0.5 ml) and Ni-Al (0.4 g)
catalyst for six hours . The mixture was filtered hot through
celite and the bed washed with methanol. The washings were
concentrated in vacuo. The concentrate was basified with 2N NaOH to
pH 9 and extracted with ethyl acetate. The ethyl acetate layer was
dried over anhydrous Na.sub.2SO.sub.4 and purified by flash column
chromatography. (0-15%) methanol/ethyl acetate gave 80% (70.10 mg)
of pure white product; mp 220-221.degree. C.; IR (KBr) 3222, 3047,
1656, 1575, 1439, 1289, 1110, 1058, 816; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 1.79 (s,3H), 3.29 (t,
2H), 3.54-3.64 (q, J=6.66, 12.64, 2H), 7.97-8.09 (m,2H), 8.19
(t,1H), 8.38 (s, 1H), 10.17 (s,1H); .sup.13C (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 22.69, 27.91, 36.75, 114.89, 116.86, 125.84,
131.67, 133.81, 135.33, 155.88, 170.13, 192.47; Anal. calculated
for C.sub.12H.sub.13N.sub.3O: C; 62.32, H; 5.67, N; 18.17 found C;
62.10, H; 5.7, N; 17.92.
Example 10
5-Phenyl-2-[2'-[2"-(2-methoxyethyl)benzimidazol-5"-yl]benzimidazol-5'-yl]b-
enzimidazole (13)
[0111] 5-Phenyl-2-[3,4-diaminophenyl]benzimidazole (0.17 mg, 0.58
mmol) and 5-formyl-2-(2-methoxyethyl)benzimidazole (120 mg, 0.59
mmol) were heated together in of nitrobenzene (5 ml) for 15 hours
at 145.degree. C. Nitrobenzene was removed with a Kugelrohr and the
compound was loaded onto a column. (1-12%) methanol/ethyl acetate
gave 80% pure yellowish product; mp>280.degree. C.; IR (KBr)
3157, 2933, 1629, 1551, 1441, 1385, 1288, 1108, 818, 760, 698;
.sup.1H NMR (DMSO-d.sub.6+3drops CF.sub.3COOH) .delta. 3.33 (s,3H),
3.45 (t,2H), 3.89 (t,2H), 7.45-7.59 (m,3H), 7.79 (d,2H,J=7.04),
7.86-7.99 (m,2H), 8.06-8.14 (m,3H), 8.26-8.31 (dd,1H, J=1.34, 8.4),
8.44-8.49 (dd,1H,J=1.02,8.82), 8.72 (s,2H); .sup.13C NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 27.71, 58.30, 68.17,
111.7, 113.69, 114.57, 115.22, 115.72, 116.19, 118.51, 123.94,
124.21, 125.26, 125.61, 127.46, 128.14, 129.36, 131.65, 131.71,
132.91, 133.49, 137.24, 138.72, 139.68, 150.15, 153.24, 155.27;
HRMS (FAB) calculated for C.sub.30H.sub.25N.sub.6O (MH.sup.+)
485.2089 found 485.2089.
[0112] The intermediate 5-formyl-2-(2-methoxyethyl)benzimidazole
was prepared as follows.
[0113] a. 5-Cyano-2(2-methoxyethyl)benzimidazole.
3,4-Diaminobenzonitrile (0.2 g, 1.5 mmol) was refluxed with (0.22
ml, 2.25 mmol) of 2-methoxypropionic acid in 4N HCl (2.5 ml) for 7
hours. After neutralization with 2N NaOH the mixture was extracted
with ethyl acetate and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuo. The product was purified on column
chromatography using (7-100%) ethyl acetate/n-hexanes to give a 50%
yield of pure white product; mp 122-123.degree. C.; IR (KBr) 2875,
2225, 1624, 1544, 1454, 1288, 1215, 1106, 824; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 3.29 (s,3H), 3.42
(t,2H), 3.84 (t,2H), 7.87-8.00 (m,2H), 8.40 (s,1H); .sup.13C NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 27.65, 58.18, 68.02,
106.79, 115.56, 118.28, 119.37, 128.95, 131.31, 134.26, 156.12;
Anal. calculated for C.sub.11H.sub.11N.sub.3.1/2 H.sub.2O: C;
62.84, H; 5.75, N; 19.97 found C; 62.71, H; 5.70, N; 18.83.
[0114] b. 5-Formyl-2-(2'-methoxyethyl)benzimidazole.
5-Cyano-2-(2-methoxyethyl)benzimidazole (0.12 g, 0.61 mmol) was
mixed in HCOOH (9 ml) and water (1 ml), to which was added Ni-Al
(0.6 g). The mixture was heated at 95.degree. C. for 5.5 hours. The
mixture was passed through a celite bed while hot and the bed was
washed with methanol and the filtrate concentrated. After
neutralization with 2N NaOH and extraction with CHCl.sub.3, the
extracts were dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was purified by column
chromatography using a gradient of 15-100% ethyl acetateln-hexanes
to give a white product in 65% yield; mp 83-84.degree. C.; IR
(Nujol) 2846, 2728, 1692, 1290, 1109,816; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 3.31(s,3H), 3.44
(t,2H), 3.86 (t,2H), 7.79-8.09 (m,2H), 8.36 (s,1H), 10.16 (s,1H);
.sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 27.73,
58.28, 68.08, 114.92, 116.86, 125.92, 131.55, 133.85, 135.19,
155.8, 192.43; Anal. calculated for
C.sub.11H.sub.12N.sub.2O.sub.21/4 H.sub.2O: C; 64.69, H; 5.92, N;
13.71 found C; 64.76, H; 6.02, N; 13.60.
Example 11
5-Bromo-2-[2'-[2"-hydroxybenzimidazol-5"-yl]benzimidazol-5'-yl]benzimidazo-
le (18).
[0115] 5-Bromo-2-[3,4-diaminophenyl]benzimidazole (33.3 mg, 0.11
mmol) and 5-formyl-2-hydroxybenzimidazole (17.7 mg, 0.11 mmol) were
heated at 145.degree. C. in nitrobenzene (3 ml) overnight.
Nitrobenzene was removed with a Kugelrohr and the compound was
purified by flash column chromatography. Elution with (5-15%)
methanol/ethyl acetate gave 23.5 mg (0.05 mmol) of yellow colored
compound in 48% yield; mp>280.degree. C.; IR (KBr) 3409, 3211,
1698, 1558, 1482, 1384, 1279; .sup.1H NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 7.30 (d,1H,J=8.34), 7.69 (dd,1H, J=1.80,
8.71), 7.82 (s,1H), 7.86-7.99 (m,2H), 8.09-8.13 (m,2H), 8.32
(dd,1H,J=1.36,8.69), 8.61 (s,1H), 11.38 (s, 1H); .sup.13C
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 108.01, 10.55, 113.92,
115.16, 115.27, 116.25, 117.19, 117.86, 121.06, 122.41, 125.24,
128.70, 130.78, 132.66, 133.16, 134.85, 134.89, 135.62, 150.42,
153.09, 155.69; HRMS (FAB) calculated for
C.sub.21H.sub.14BrN.sub.6O (MH.sup.+) 445.0412 found 445.0408.
[0116] The intermediate 5-bromo-2-[3,4-diaminophenyl]benzimidazole
was prepared as follows.
[0117] a. 4-Bromo-o-phenylenediamine. 4-Bromo-2-nitroaniline 600 mg
(2.76 mmol) was dissolved in 25 ml absolute ethanol and 2.72 g (14
mmol) SnCl.sub.2 was added. The mixture was refluxed overnight.
Ethanol was removed in vacuo and the mixture basified with 2N NaOH
to pH 11. Ether extraction, drying the ether layer over anhydrous
Na.sub.2SO.sub.4 and concentration in vacuo afforded 486 mg (2.6
mmol, 94% yield) of the crude 4-bromo-o-phenylenediamine which was
used for the next step without characterization.
[0118] b. 5-Bromo-2-[3,4-nitrophenyl]benzimidazole.
4-Bromo-o-phenylenediamine (275 mg, 1.5 mmol) and
3,4-dinitrobenzaldehyde (300 mg, 1.5 mmol) in 2 ml nitrobenzene
were heated at 145.degree. C. overnight. The nitrobenzene was
removed with a Kugelrohr. Column purification (1-10 % ethyl
acetate/n-hexanes) afforded 209 mg (0.57 mmol, 39% yield) of the
pure product; .sup.1H NMR (DMSO+3 drops CF.sub.3 COOH).delta. 7.48
(dd,1H,J=1.82,8.76), 7.7 (d,1H,8.76), 7.96 (d,1H, 1.46), 8.45
(d,1H,8.5), 8.68 (dd,1H,J=1.84,8.5), 8.93 (d,1H,1.84); .sup.13C NMR
(DMSO+3 drops CF.sub.3COOH) .delta. 116.36, 117.48, 118.66, 123.74,
127.05, 132.07, 134.49, 137.59, 140.23, 142.39, 142.81, 148.45.
[0119] c. 5-Bromo-2-[3,4-diaminophenyl]benzimidazole.
5-Bromo-2-[3,4-dinitrophenyl]benzimidazole (140 mg, 0.38 mmol) was
dissolved in 8 ml absolute ethanol. 0.8 g SnCl.sub.2 (4.2 mmol) was
added and the mixture refluxed overnight. Ethanol was removed in
vacuo and the mixture basified with 2N NaOH to pH 11. Repeated
extraction with ether, drying the ether layer over anhydrous
Na.sub.2SO.sub.4 and concentration in vacuo yielded (0.11 g, 0.37
mmol) of the crude diamine in 98% yield. The diamine was used
without further purification; .sup.1H NMR (DMSO-d.sub.6) .delta.
6.63 (d,1H,J=8.1), 7.26 (m,2H), 7.39 (m,2H), 6.65
(d,1H,J=1.76).
[0120] The intermediate 5-formyl-2-hydroxybenzimidazole was
prepared as follows.
[0121] d. 5-Cyano-2-hydroxybenzimidazole. 3,4-Diaminobenzonitrile
(0.5 g, 3.8 mmol) and urea (0.25 g, 4.2 mmol) were heated together
in 1 ml DMF at 145-150.degree. C. for 9 h. The DMF was concentrated
in vacuo and the mixture was suspended in water and extracted with
ethyl acetate. Purification by column chromatography (elution with
60/40 ethyl acetate/n-hexanes) provided (0.53 g, 3.3 mmol) a yield
of 89% of the product; IR (KBr) 3533, 3209, 2224, 1732, 1482, 1281;
.sup.1H NMR (DMSO-d.sub.6+3 drops CF.sub.3 COOH) .delta. 7.1
(d,1H,J=8.06), 7.09-7.41 (m,2H), 11.06 (s,1H), 11.19 (s,1H);
.sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta.
102.52,109.36, 111.61, 120.04, 126.03, 130.18, 133.93, 155.41.
[0122] e. 5-Formyl-2-hydroxybenzimidazole.
5-Cyano-2-hydroxybenzimidazole (0.5 g, 3.14 mmol) was mixed with
Ni-Al catalyst (3 g), HCOOH (45 ml) and water (3 ml) and heated to
reflux for 12 hrs. The hot mixture was filtered through a bed of
celite and the filtrate concentrated in vacuo. pH of the mixture
was adjusted to 9 with 2N NaOH and extraction was carried out with
ethyl acetate. The ethyl acetate layer was dried over anhydrous
Na.sub.2SO.sub.4 and concentrated in vacua. Column chromatographic
purification, and elution with 65/35 ethyl acetate/n-hexanes
provided 0.4 g (2.4 mmol, 76% yield) of the final compound; IR
(KBr) 3261, 3160, 1677, 1631, 1477, 1282, 711; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3 COOH) .delta. 7.11 (d,1H,J=8.0),
7.41 (s,1H), 7.57 (dd,1H,J=1.54,8.0), 9.86 (s,1H), 10.99 (s,1H),
11.17 (s,1H); .sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3 COOH)
.delta. 108.15, 108.67, 125.37, 130.1, 130.42, 135.64, 155.68,
191.99.
Example 12
5,6-Dibromo-2-[2'-(2"-trifluoromethylbenzimidazol-5"-yl)benzimidazol-5'yl]-
benzimidazole (22).
[0123] 4,5-Dibromo-o-phenylenediamine (60 mg, 0.22 mmol) and
5-formyl-2-(2'-trifluoromethylbenzimidazol-5'-yl)benzimidazole (73
mg, 0.22 mmol) were heated in 10 ml nitrobenzene overnight at
150.degree. C. The nitrobenzene was removed witha Kugelrohr and the
mixture loaded on a column. Elution with 90/10 ethyl
acetate/n-hexanes yielded 25 mg (20% yield, 0.05 mmol) of compound;
mp>260.degree. C.; .sup.1H NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 8.02-8.09 (m,2H), 8.14 (s,2H), 8.26-8.29
(m,2H), 8.57 (s,1H), 8.69 (s,1H).
[0124] The intermediate
5-formyl-2-(2'-trifluoromethylbenzimidazol-5'-yl)b- enzimidazole
was prepared as follows.
[0125] a.
5-Cyano-2-(2'-trifluoromethylbenzimidazol-5'-yl)benzimidazole.
5-formyl-2-trifluoromethylbenzimidazole (1.5 g, 7 mmol) was mixed
with 0.93 g of 3,4-diaminobenzonitrile (7 mmol) and 15 ml
nitrobenzene. The mixture was heated overnight at 145.degree. C.
Nitrobenzene was removed with a Kugelrohr and the mixture was
loaded onto a column. Elution with 40/60 ethyl acetate/n-hexanes
gave 0.9 g (2.75 mmol) of the compound in 40% yield;
mp>260.degree. C.; IR (KBr) 3254, 2916, 2223, 1614, 1175;
.sup.1H NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 7.74
(dd,1H,J=1.38,8.4), 7.88 (d,1H,J=8.14), 7.98 (d,1H,J=8.78),
8.25-8.03 (m,2H), 8.63 (s,1H); .sup.13C NMR (DMSO-d.sub.6 +3 drops
CF.sub.3COOH) .delta. 105.62, 115.78, 116.35, 120.26, 123.06,
123.82, 127.18, 137.35, 139.65, 142.17, 142.96, 154.13.
[0126] b.
5-Formyl-2-(2'-trifluoromethylbenzimidazol-5'-yl)benzimidazole.
5-Cyano-2-(2'-trifluoromethylbenzimidlazol-5'-yl)benzimidazole
(0.18 g, 0.54 mmol) was refluxed in 8 ml HCOOH with 2.5 ml water
and 0.6 g Ni-Al catalyst for 7 hours at 110.degree. C. The hot
mixture was filtered through celite bed and the filtrate
concentrated and its pH adjusted to 9.0 with 2N NaOH. Repeated
extraction with ethyl acetate, drying the organic layer with
anhydrous Na.sub.2SO.sub.4 and concentration in vacuo gave the
crude product. Purification by column chromatography (elution with
1% methanol/ethyl acetate) yielded 87 mg (0.26 mmol, 49% yield) of
the pure aldehyde; mp>260.degree. C.; IR (KBr) 3411, 2966, 1679,
1310, 1153; .sup.1H NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta.
7.96-8.06 (m,3H), 8.30-8.32 (m,2H), 8.57 (s,1H), 10.15 (s,1H);
.sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 106.77,
112.51, 113.28, 117.79, 117.84, 118.28, 121.78, 123.96, 125.02,
132.98, 135.79, 139.62, 142.48, 153.51, 192.65.
[0127] The intermediate 4,5-dibromo-1,2-phenylenediamine was
prepared as follows, using a modification of the procedures
described in U.S. Pat. No. 4,264,600.
[0128] c. 3,4-Dibromoaniline. 3-Bromoaniline (3 g, 17.4 mmol) was
dissolved in 15 ml CH.sub.2Cl.sub.2 and the reaction mixture was
cooled to -10.degree. C. in an ice-salt bath.
2,4,4,6-Tetrabromo-2,5-cyclohexadi- enone, (9.29 g, 0.02 mmol) was
added in small portions with constant stirring. The reaction
mixture was stirred for a period of 7 hours at 0.degree. C. after
which the reaction was quenched with 2N NaOH (10 ml). The aqueous
layer was extracted with CH.sub.2Cl.sub.2 and the organic layer was
dried over anhydrous Na.sub.2SO.sub.4. The CH.sub.2Cl.sub.2 layer
was concentrated in vacuo and purified by column chromatography.
Elution with 0-3% ethyl acetate/n-hexanes yielded 3.2 g (12.7 mmol,
73%) of pure product; mp 80-81.degree. C.; IR (KBr) 3406, 3318,
3210, 1583, 1464, 1287, 1108, 860, 668; .sup.1H NMR (CDCl.sub.3)
.delta. 6.49 (dd,1H,J=2.7,8.6), 6.7 (d,1H,J=2.64), 7.32
(d,1H,J=8.6); .sup.13C NMR(CDCl.sub.3) .delta. 112.6, 116, 120.1,
125.45, 134.26, 147.08.
[0129] d. 3,4-Dibromoacetanilide. 3,4-Dibromoaniline (3.2 g, 12.7
mmol) was mixed with 13 ml acetic anhydride and heated at
110.degree. C. for 2 hours after which it was poured over ice cold
water. The aqueous layer was extracted with ethyl acetate, 0.1N HCI
and 1N NaOH, and the organic layer was dried over anhydrous
Na.sub.2SO.sub.4. The organic layer was concentrated in vacuo and
purified by column chromatography. Elution with 1-50% ethyl
acetate/n-hexanes yielded 2.6 g (9 mmol, 71% yield) of the
acetanilide: mp 91-93.degree. C.; IR (KBr) 3293, 3172, 1668,
1589,1552, 1522, 1373, 813; .sup.1H NMR (CDCl.sub.3) .delta.
2.17(s,3H), 7.33 (dd,1H,J=2.4,8.66), 7.52 (d,1H,J=8.68), 7.87
(d,1H, J=2.4); .sup.13C NMR (CDCl.sub.3) .delta. 25.07, 119.64,
120.35, 125.04, 125.38, 134.13, 138.40, 168.96.
[0130] e. 3,4-Dibromo-6-nitroacetanilide. 3,4-Dibromoacetanilide
(2.64 g, 9 mmol) was dissolved in 8 ml conc. H.sub.2SO.sub.4
(chilled in ice-salt bath) at a temperature of 0.degree. C. To this
solution was added 1 ml chilled nitrating mixture (equal volumes of
conc. H.sub.2SO.sub.4 and fuming HNO.sub.3) and the mixture stirred
for 20 min. The reaction mixture was poured into ice water and
neutralized with ammonium hydroxide to pH of 9.0. The yellow
precipitate was filtered and purified by column chromatography.
Elution with 1-4% ethyl acetate/n-hexanes yielded 2.1 g (6.15 mmol,
68%) of the 3,4-dibromo-6-nitroacetanilide: mp 141-142.degree. C.;
IR (KBr) 3340, 3128, 1693, 1569, 1479, 1332, 1268; .sup.1H NMR
(CDCl.sub.3) .delta. 2.3 (s,3H), 8.46 (s, 1H), 9.22 (s,1 H);
.sup.13C NMR (CDCl.sub.3) .delta. 26.14, 118.67, 126.86, 130.86,
134.51, 134.59, 136.98, 169.40.
[0131] f. 3,4-Dibromo-6-nitroaniline.
3,4-Dibromo-6-nitroacetanilide (2.1 g, 6.15 mmol) was heated to
refluxing temperature for 2 hours with 40 ml of 6N HCI. The
reaction mixture was then poured over an ice-water mixture and
stirred. The pH of the solution was adjusted to 9.0 with NaOH. The
resulting bright yellow precipitate was filtered, washed with cold
water and dried to yield (1.78 g, 6 mmol, 98%) of pure product; mp
204-205.degree. C.; IR (KBr) 3475, 3355, 3097, 1612, 1478, 1241,
1122, 914; .sup.1H NMR (CDCl.sub.3) .delta. 6.05 (brs,2H), 7.18
(s,1H), 8.37 (s,1H); .sup.13C NMR (CDCl.sub.3) .delta. 111.23,
123.22, 129.34, 130.59, 133.61, 142.36.
[0132] g. 4,5-Dibromo-1,2-phenylenediamine.
3,4-Dibromo-6-nitroaniline (189 mg, 0.64 mmol) was dissolved in 8
ml anhydrous ethanol to which was added about 400 mg Raney Nickel.
The hydrogenation apparatus was at a hydrogen pressure 50 psi.
After 45 min. the deep yellow color originally present was
completely discharged indicating complete reduction of the nitro to
the amine groups. The reaction mixture was filtered through celite
and the bed was washed with methanol. The methanol was concentrated
in vacuo to give 135 mg of the crude diamine in 82% yield. The
crude diamine was used as such without further purification;
.sup.1H NMR (CDCl.sub.3) .delta. 3.37 (br,4H), 6.93 (s,2H);
.sup.13CNMR (CDCl.sub.3) .delta. 113.98, 120.95, 135.84.
Example 13
5-Phenyl-6methoxy-2-[2'(benzimidazol-5"-yl)benzimidazol-5'-yl]benzimidazol-
e (15).
[0133] 4-Methoxy-5-phenyl-1,2 phenylenediamine (139 mg, 0.65 mmol)
and 5-formyl-2-(benzimidazol-5'-yl)benzimidazole (170 mg, 0.65
mmol) were heated together in nitrobenzene (5 ml) overnight at
145.degree. C. Nitrobenzene was removed with a Kugelrohr and the
compound was loaded onto a column. (1-10%) methanol/ethyl acetate
gave 120 mg (41%) of pure product; mp>280.degree. C.; IR (KBr)
3298, 3050, 2987, 1630, 1541, 1438, 1283; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 3.92 (s,3H), 7.39-7.57
(m,6H), 7.72 (s,1H), 8.11-8.15 (m,1H), 8.19-8.27 (m,2H), 8.47
(dd,1H,J=1.46,8.07), 8.64 (s,1H), 8.76 (s,1H), 9.75 (s,1H);
.sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 56.40,
95.73, 114.62, 115.18, 116.02, 116.15, 119.03, 119.07, 125.59,
126.29, 127.63, 127.67, 128.34, 129.79, 130.69, 131.54, 132.42,
133.38, 137.59, 148.81, 152.84, 156.82; HRMS (FAB) calculated for
C.sub.28H.sub.21N.sub.6O (MH.sup.+) 457.1777, found 457.1770.
[0134] The intermediate 4-methoxy-5-phenyl-1,2 phenylenediamine was
prepared as follows.
[0135] a. 4-Methoxy-2-nitro5-phenylaniline.
3-Bromo4-methoxy-6-nitroanilin- e (400 mg, 1.62 mmol) was dissolved
in DME (20 ml). Tetrakispalladium triphenyl phosphine (94 mg, 0.08
mmol) served as the catalyst. To this mixture was added phenyl
boronic acid(300 mg, 1.01 mmol) and 2M Na.sub.2CO.sub.3 (1.8 ml)
and the mixture refluxed at 90.degree. C. overnight. The reaction
mixture was concentrated in vacuo and purified by flash column
chromatography. (1-10%) ethyl acetate/n-hexanes gave 280 mg (1.14
mmol) of pure product in 71% yield; IR (KBr) 3435, 3328, 2933,
1570, 1480, 1227, 1029, 692; .sup.1H NMR (CDCl.sub.3) .delta. 3.80
(s,3H), 5.94 (br,2H), 6.79 (s,1H), 7.39-7.64 (m,5H), 7.64 (s,1H);
.sup.13C NMR (CDCl.sub.3) .delta. 56.63, 106.76, 121.26, 128.69,
128.72, 128.76, 128.79, 128.92, 129.71, 136.69, 140.24, 141.42,
148.65; MS (FAB) calculated for C.sub.13H.sub.12N.sub.2O.sub.3
244.0848, found 244.0847.
[0136] b. 4-Methoxy-5-phenyl-1,2 phenylenediamine.
4-Methoxy-2-nitro-5-phe- nyl-aniline (165 mg, 0.68 mmol) in ethyl
acetate (20 ml) was reduced using 20 mg of 10% Pd/C as the
catalyst, overnight. The mixture was filtered through celite bed
and the bed was washed with methanol. All the washings were
concentrated in vacuo to yield 139 mg (96% yield) of the crude
diamine which was used without further purification; .sup.1H NMR
(CDCl.sub.3) .delta. 3.49 (brs,4H), 3.71 (s,3H), 6.42 (s, 1H), 6.74
(s,1H), 7.26-7.52 (m,5H); .sup.13C NMR (CDCl.sub.3) .delta. 56.81,
101.77, 120.93, 127.53, 128.39, 128.45, 128.49, 129.82, 129.96,
136.64, 139.17, 142.37, 153.73.
[0137] The intermediate 3-bromo-4-methoxy-6-nitro-aniline used in
sub-part a is commercially available (Aldrich Chemical
Company).
[0138] The intermediate
5-fornyl-2-(benzimidazol-5'-yl)benzimidazole was prepared as
described by Sun et al. J. Med. Chem. 1995, 38, 3638-3644.
Example 14
5,6-Diphenyl-2-[2'(benzimidazol-5"-yl)benzimidazol-5'-yl]benzimidazole
(16).
[0139] 4,5-Diphenyl-1,2-phenylenediamine (151 mg, 0.58 mmol) and
5-formyl-2-(benzimidazol-5'-yl)benzimidazole (152 mg, 0.58 mmol)
were heated in nitrobenzene (4 ml) overnight at 145.degree. C.
under nitrogen. The nitrobenzene was removed with a Kugelrohr.
Compound was purified by flash column chromatography. Elution with
(2-10%) methanol/ethyl acetate gave 108 mg (0.22 mmol) of pure
yellow compound in 37% yield. mp>280.degree. C.; IR (KBr) 3399,
3059, 1629, 1551, 1441, 1292; .sup.1H NMR (DMSO-d.sub.6 +3 drops
CF.sub.3COOH) .delta. 7.17-7.33 (m,10H), 7.84 (s,2H),8.06-8.25
(m,3H), 8.49 (dd,1H,J=1.18,8.89), 8.66 (s,1H), 8.75 (s,1H), 9.69
(s,1H); .sup.13C NMR (DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta.
113.94, 115.24, 115.84, 116.15, 116.32, 117.89, 123.34, 126.32,
126.66, 127.86, 128.38, 128.42, 128.46, 130.05, 131.77, 131.89,
133.03, 138.87, 138.97, 140.62, 141.19, 160.97, 153.71; HRMS (FAB)
calculated for C.sub.33H.sub.23N.sub.6 (MH.sup.+) 503.1984 found
503.1989.
[0140] The intermediate 4,5-diphenyl-1,2-phenylenediamine was
prepared as follows.
[0141] a. 4,5-Diphenyl-2-nitroaniline. 3,4Dibromo-6-nitroaniline
(332 mg, 1.12 mmol) was dissolved in DME (20 ml).
Tetrakis(triphenylphosphine)pall- adium (65 mg, 0.06 mmol), phenyl
boronic acid (200 mg, 1.64 mmol), and 2M Na.sub.2CO.sub.3 (10 ml)
were added to the reaction mixture and refluxed at 90.degree. C.
overnight. The reaction mixture was concentrated in vacuo and
loaded onto a column. (1-3%) ethyl acetate/n-hexanes afforded 259
mg of pure yellow compound in 80% yield; mp 139-141.degree. C.; IR
(KBr) 3476, 3363, 2924, 1621, 1476, 1263, 1089; .sup.1H NMR
(CDCl.sub.3) .delta. 6.11 (brs,2H), 6.86 (s,1H), 7.04-7.09 (m,5H),
7.15-7.26 (m,5H), 8.21 (s,1H); .sup.13C NMR (CDCl.sub.3) .delta.
120.56, 127.22, 127.66, 128.20, 128.26, 128.52, 128.53, 129.31,
129.39, 129.47, 129.77, 130.14, 130.93, 131.79, 139.85, 139.94,
144.08, 149.03.
[0142] b. 4,5-Diphenyl-1,2-phenylenediamine.
4,5-Diphenyl-2-nitroaniline (200 mg, 0.69 mmol) in ethanol (50 ml)
was reduced using 40 mg 10% Pd/C. Hydrogenation was carried out at
40 psi pressure for 10 hours. The mixture was filtered through
celite and the bed washed with methanol. The methanol layer was
concentrated in vacuo to yield 151 mg of the crude diamine in 84%
yield. The crude diamine was used as such without purification.
.sup.1H NMR (CD.sub.3OD) .delta. 6.78 (s,2H), 7.01-7.16 (m,10H);
.sup.13C NMR (CD.sub.3OD) .delta. 118.08, 121.23, 127.17, 127.66,
130.44, 131.76, 138.32.
Example 15
5,6-Dibromo-2-[2'-(benzimidazol-5"-yl)benzimidazol-5'yl]
benzimidazole (20).
[0143] 4,5-Dibromo-1,2-phenylenediamine (128 mg, 0.48 mmol) and
5-formyl-2-(benzimidazol-5'-yl)benzimidazole (126 mg, 0.48 mmol)
were heated in nitrobenzene (6 ml) at 145.degree. C. overnight. The
nitrobenzene was removed with a Kugelrohr and the mixture loaded on
a column for purification. Elution with (1-10%) methanol/ethyl
acetate gave 0.1 gm (41%) of pure compouind; mp>260.degree. C.;
IR(KBr) 3405, 3198, 1626, 1544, 1385, 1292; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 8.03-8.13 (m,2H), 8.17
(s,2H), 8.25 (d,1H,J=9.16), 8.42 (d,1H,J=8.64), 8.59 (s,1H), 8.74
(s,1H), 9.75 (s,1H); .sup.13C NMR (DMSO-d.sub.6+3 drops
CF.sub.3COOH) .delta. 114.73, 114.96, 115.77, 115.99, 118.93,
119.61, 120.73, 123.29, 124.47, 125.56, 131.49, 133.47, 134.77,
135.79, 137.95, 151.93, 152.31; HRMS (FAB) calculated for
C.sub.21H.sub.13Br.sub.2N.sub.6 (MH.sup.+) 506.9568 found
506.9574.
[0144] The intermediate 4,5-dibromo-1,2-phenylenediamine was
prepared as described at Example 12c-g.
Example 16
5-Bromo-6-metboxy-2-[2'-(benzimidazol-5"-yl)benzimidazol-5'yl]benzimidazol-
e (21)
[0145] In nitrobenzene (4ml),
4-bromo-5-methoxy-1,2-phenylenediamine (123 mg, 0.57 mmol) and
5-formyl-2-(benzimidazol-5'-yl)benzimidazole (150 mg, 0.57 mmol)
were heated at 145.degree. C. overnight under nitrogen.
Nitrobenzene was removed with a Kugelrohr. Chromatographic
separation with (1-10%) methanol/ethyl acetate afforded 104 mg
(40%) of pure compound; mp >280.degree. C.; .sup.1H NMR
(DMSO-d.sub.6+3 drops CF.sub.3COOH) .delta. 3.91 (s,3H), 7.17-7.29
(m,1H), 7.78 (d,1H,J=8.84), 8.00-8.13 (m,3H), 8.49-8.53 (m,1H),
8.66 (s,1H), 8.79 (s,1H), 9.69 (s,1H); .sup.13C NMR (DMSO-d.sub.6+3
drops CF.sub.3COOH) .delta. 56.16, 96.43, 114.06, 114.97, 115.55,
115.79, 115.89, 116.35, 117.97, 118.42, 123.11, 125.30, 125.58,
126.33, 127.43, 131.85, 133.05, 133.11, 138.39, 140.84, 149.04,
153.44; HRMS (FAB) calculated for C.sub.22H.sub.16BrN.sub- .6O
(MH.sup.+)
[0146] The intermediate 4-bromo-5-methoxy-1,2-phenylenediamine was
prepared as follows.
[0147] a. 4-Bromo5-methoxy-1,2-phenylenediamine.
3-Bromo4-methoxy-6-nitroa- niline (150 mg, 0.61 mmol) (Aldrich
Chemical Company) was dissolved in ethanol (10 ml) and
hydrogenation was carried out using 350 mg Raney Nickel for 30 min.
The reaction mixture was filtered through celite bed and washed
with methanol. The methanolic layer was dried in vacuo to give 127
mg of the crude diamnine in 97.40% yield, which was used as such
withoult purification; .sup.1H NMR (CDCl.sub.3) .delta. 3.20
(brs,2H), 3.39 (brs,2H), 3.8 (s,3H), 8.36 (s,1H), 6.89 (s,1H);
.sup.13C NMR (CDCl.sub.3) .delta. 57.35, 100.38, 102.27, 122.52,
128.69, 136.68, 150.94.
Example 17
[0148] The following illustrate representative pharmaceutical
dosage forms, containing a compound of formula I (`Compound X`),
for therapeutic or prophylactic use in humans.
1 (i) Tablet 1 mg/tablet `Compound X` 100.0 Lactose 77.5 Povidone
15.0 Croscarmellose sodium 12.0 Microcrystalline cellulose 92.5
Magnesium stearate 3.0 300.0 (ii) Tablet 2 mg/tablet `Compound X`
20.0 Microcrystalline cellulose 410.0 Starch 50.0 Sodium starch
glycolate 15.0 Magnesium stearate 5.0 500.0 (iii) Capsule
mg/capsule `Compound X` 10.0 Colloidal silicon dioxide 1.5 Lactose
465.5 Pregelatinized starch 120.0 Magnesium stearate 3.0 600.0 (iv)
Injection 1 (1 mg/ml) mg/ml `Compound X` (free acid form) 1.0
Dibasic sodium phosphate 12.0 Monobasic sodium phosphate 0.7 Sodium
chloride 4.5 1.0N Sodium hydroxide solution q.s. (pH adjustment to
7.0-7.5) Water for injection q.s. ad 1 mL (v) Injection 2 (10
mg/ml) mg/ml `Compound X` (free acid form) 10.0 Monobasic sodium
phosphate 0.3 Dibasic sodium phosphate 1.1 Polyethylene glycol 400
200.0 01N Sodium hydroxide solution q.s. (pH adjustment to 7.0-7.5)
Water for injection q.s. ad 1 mL (vi) Aerosol mg/can `Compound X`
20.0 Oleic acid 10.0 Trichioromonofluoromethane 5,000.0
Dichiorodifluoromethane 10,000.0 Dichlorotetrafluoroethane
5,000.0
[0149] The above formulations may be obtained by conventional
procedures well known in the pharmaceutical art.
[0150] All publications, patents, and patent documents are
incorporated by reference herein, as though individually
incorporated by reference. The invention has been described with
reference to various specific and preferred embodiments and
techniques. However, it should be understood that many variations
and modifications may be made while remaining within the spirit and
scope of the invention.
* * * * *