U.S. patent application number 11/339296 was filed with the patent office on 2007-01-18 for nitro and amino substituted heterocycles as topoisomerase i targeting agents.
This patent application is currently assigned to Rutgers, The State University of New Jersey. Invention is credited to Edmond J. LaVoie, Leroy F. Liu, Sudhir K. Singh.
Application Number | 20070015751 11/339296 |
Document ID | / |
Family ID | 31715798 |
Filed Date | 2007-01-18 |
United States Patent
Application |
20070015751 |
Kind Code |
A1 |
LaVoie; Edmond J. ; et
al. |
January 18, 2007 |
Nitro and amino substituted heterocycles as topoisomerase I
targeting agents
Abstract
The invention provides compounds of formula I: ##STR1## wherein:
R.sub.1-R.sub.5,"a" and X have any of the meanings defined in the
specification and their pharmaceutically acceptable salts. The
invention also provides pharmaceutical compositions comprising a
compound of formula I, processes for preparing compounds of formula
I, intermediates useful for preparing compounds of formula I, and
therapeutic methods for treating cancer using compounds of formula
I.
Inventors: |
LaVoie; Edmond J.;
(Princeton Junction, NJ) ; Singh; Sudhir K.;
(Bangalore, IN) ; Liu; Leroy F.; (Bridgewater,
NJ) |
Correspondence
Address: |
VIKSNINS HARRIS & PADYS PLLP
P.O. BOX 111098
ST. PAUL
MN
55111-1098
US
|
Assignee: |
Rutgers, The State University of
New Jersey
|
Family ID: |
31715798 |
Appl. No.: |
11/339296 |
Filed: |
January 25, 2006 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
10638943 |
Aug 11, 2003 |
6992088 |
|
|
11339296 |
Jan 25, 2006 |
|
|
|
60402166 |
Aug 9, 2002 |
|
|
|
Current U.S.
Class: |
514/233.2 ;
514/283; 514/284; 544/125; 546/48; 546/61 |
Current CPC
Class: |
A61P 35/00 20180101;
C07D 471/04 20130101; C07D 221/18 20130101 |
Class at
Publication: |
514/233.2 ;
514/283; 514/284; 544/125; 546/048; 546/061 |
International
Class: |
A61K 31/5377 20070101
A61K031/5377; A61K 31/4741 20070101 A61K031/4741; A61K 31/473
20070101 A61K031/473; C07D 491/04 20070101 C07D491/04; C07D 221/18
20070101 C07D221/18 |
Goverment Interests
GOVERNMENT FUNDING
[0002] The invention described herein was made with government
support under Grant Numbers CA39662 and CA077433 from the National
Cancer Institute. The United States Government has certain rights
in the invention.
Claims
1. A compound of formula I: ##STR4## wherein: one of R.sub.1 and
R.sub.2 is nitro or NR.sub.aR.sub.b; the other of R.sub.1 and
R.sub.2 is hydrogen, (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, NR.sub.aR.sub.b, COOR.sub.c, or
OR.sub.d; and R.sub.3 and R.sub.4 are each independently hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
NR.sub.aR.sub.b , COOR.sub.c, or OR.sub.d, or R.sub.3 and R.sub.4
taken together are methylenedioxy, 1,2-ethylenedioxy, or
1,3-propylenedioxy; or 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,
NR.sub.aR.sub.b, COOR.sub.c, or OR.sub.dor R.sub.1 and R.sub.2
taken together are methylenedioxy, 1,2-ethylenedioxy, or 1,3-
propylenedioxy; one of R.sub.3 and R.sub.4 is nitro or
NR.sub.aR.sub.b; and the other of R.sub.3 and R.sub.4 is hydrogen,
(C-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl, NR.sub.aR.sub.b,
COOR.sub.c, or OR.sub.d; R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one or more solubilizing groups; X is two
hydrogens, =O, =S, or =Nhd e; the bond marked"a" is a single bond
or a double bond: R.sub.a and R.sub.b are each independently
hydrogen or (C.sub.1-C.sub.6)alkyl, or R.sub.a and R.sub.b together
with the nitrogen to which they are attached form a pyrrolidino,
piperidino or morpholino ring; each R.sub.c is hydrohgen,
(C.sub.1-C.sub.6)alkyl, aryl, or aryl(C.sub.1-C.sub.6)alkyl; each
R.sub.d is hydrogen, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkanoyl, aryl, or aryl(C.sub.1-C.sub.6)alkyl; and
R.sub.e is hydrogen, (C.sub.1-C.sub.6)alkyl, aryl, or
aryl(C.sub.1-C.sub.6)alkyl; or a pharmaceutically acceptable salt
thereof. 2-59. (canceled)
60. A method of inhibiting cancer cell growth, comprising
administering to a mammal afflicted with cancer, an amount of a
compound as described in claim 1, effective to inhibit the growth
of said cancer cells.
61. (canceled)
62. A method of producing an antibacterial effect in a mammal in
need of such treatment comprising administering to the mammal, an
amount of a compound as described in claim 1, effective to provide
an antibacterial effect.
63. (canceled)
Description
PRIORITY OF INVENTION
[0001] This application claims priority to United States
Provisional Patent Application Number 60/402166, filed 09 Aug.
2002.
BACKGROUND OF THE INVENTION
[0003] DNA-topoisomerases are enzymes which are present in the
nuclei of cells where they catalyze the breaking and rejoining of
DNA strands, which control the topological state of DNA. Recent
studies also suggest that topoisomerases are also 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 that 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.
[0004] 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). 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. Other topoisomerase poisons have been identified including
certain benzo[i]phenanthridine and cinnoline compounds (see LaVoie
et al., U.S. Pat. No. 6,140,328 and WO 01/32631).
[0005] International Patent Application Publication Number 00/21537
discusses certain specific indenoisoquinolines that are reported to
have antineoplastic activity.
[0006] Despite these reports there is currently a need for
additional agents that are useful for treating cancer.
SUMMARY OF THE INVENTION
[0007] Applicant has discovered compounds that show inhibitory
activity against topoisomerase I and/or topoisomerase II, and
compounds that are effective cytotoxic agents against cancer cells,
including drug-resistant cancer cells. In particular, Applicant has
discovered that substitution of a nitro, amino, or a substituted
amino group for either one or more of the methoxyl groups or the
methylenedioxy groups of tetracyclic topoisomerase I-targeting
agents unexpectedly provides compounds with high and potent
cytotoxic activity. Accordingly, the invention provides a compound
of the invention which is a compound of formula I: ##STR2##
wherein:
[0008]
[0009] one of R.sub.1 and R.sub.2 is nitro or NR.sub.aR.sub.b; the
other of R.sub.1 and R.sub.2 is hydrogen, (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, NR.sub.aR.sub.b, COOR.sub.c, or
OR.sub.d; and R.sub.3 and R.sub.4 are each independently hydrogen,
(Cl-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl, NR.sub.aR.sub.b,
COOR.sub.c, or OR.sub.d, or R.sub.3 and R.sub.4 taken together are
methylenedioxy, 1,2-ethylenedioxy, or 1,3-propylenedioxy; or
[0010] 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,
NR.sub.aR.sub.b, COOR.sub.c, or OR.sub.d, or R.sub.1 and R.sub.2
taken together are methylenedioxy, 1,2-ethylenedioxy, or
1,3-propylenedioxy; one of R.sub.3 and R.sub.4 is nitro or
NR.sub.aR.sub.b; and the other of R.sub.3 and R.sub.4 is hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
NR.sub.aR.sub.b, COOR.sub.c, or OR.sub.d;
[0011] R.sub.5 is (C.sub.1-C.sub.6)alkyl substituted with one or
more solubilizing groups; [0012] X is two hydrogens, =O, =S, or
=NR.sub.c; [0013] the bond marked"a" is a single bond or a double
bond: [0014] R.sub.a and R.sub.b are each independently hydrogen or
(C.sub.1-C.sub.6)alkyl, or R.sub.a and R.sub.b together with the
nitrogen to which they are attached form a pyrrolidino, piperidino
or morpholino ring; [0015] each R.sub.c is hydrogen,
C.sub.1-C6)alkyl, aryl, or aryl(C.sub.1-C.sub.6)alkyl; [0016] each
R.sub.d is hydrogen, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkanoyl, aryl, or aryl(C.sub.1-C.sub.6)alkyl; and
[0017] R.sub.e is hydrogen, (C.sub.1-C.sub.6)alkyl, aryl, or
aryl(C.sub.1-C.sub.6)alkyl; [0018] or a pharmaceutically acceptable
salt thereof.
[0019] The invention also provides a pharmaceutical composition
comprising a effective amount of a compound of the invention in
combination with a pharmaceutically acceptable diluent or
carrier.
[0020] The invention also provides a method of inhibiting cancer
cell growth, comprising administering to a mammal afflicted with
cancer, an amount of a compound of the invention, effective to
inhibit the growth of said cancer cells.
[0021] The invention also provides a method comprising inhibiting
cancer cell growth by contacting said cancer cell in vitro or in
vivo with an amount of a compound of the invention, effective to
inhibit the growth of said cancer cell.
[0022] The invention also provides a compound of the invention for
use in medical therapy, preferably for use in treating cancer, for
example, solid tumors, as well as the use of a compound of the
invention for the manufacture of a medicament useful for the
treatment of cancer, for example, solid tumors.
[0023] 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
[0024] FIG. 1 illustrates the synthesis of representative compounds
of formula I (4a, 4b, 5a, and 5b).
[0025] FIG. 2 illustrates the synthesis of representative compounds
of formula I (9a, 9b, 10a, and 10b).
[0026] FIG. 3 illustrates the synthesis of representative compounds
of formula I (13a, 13b, 14a, 14b, 15a, 15b, 16a, and 16b).
[0027] FIG. 4 illustrates the synthesis of representative compounds
of formula I (19a, 19b, 20a, 20b, 21a, 21b, 22a, and 22b)
[0028] FIG. 5 illustrates the synthesis of a representative
compounds of formula I (Compound A and Compound B)
DETAILED DESCRIPTION
[0029] The following definitions are used, unless otherwise
described. [0030] "(C.sub.1-C.sub.6)alkyl" denotes both straight
and branched carbon chains with 1, 2, 3, 4, 5, or 6, carbon atoms,
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. [0031]
"(C.sub.3-C.sub.6)cycloalkyl" denotes a carbocyclic ring with 3, 4,
5, or 6, carbon atoms. [0032] "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. Examples of aryl
include phenyl, indenyl, and naphthyl. [0033]
"Aryl(C.sub.1-C.sub.6)alkyl" refers to a group of the formula
aryl--(C.sub.1-C.sub.6)alkyl-, where aryl and
(C.sub.1-C.sub.6)alkyl are as defined herein. 1. "Solubilizing
group (R.sub.z)" is a substituent that increases the water
solubility of the compound of formula I compared to the
corresponding compound lacking the R.sub.z substituent (i.e.
wherein R.sub.z is hydrogen). Examples of solubilizing groups
include (C.sub.1-C.sub.6)alkoxycarbonyl (e.g. -CO.sub.2Me), cyano,
halo, hydroxy, mercapto, oxo (=O), carboxy (COOH), nitro,
pyrrolidinyl, piperidinyl, imidazolidinyl, imidazolinyl,
piperazinyl, morpholinyl, thiomorpholinyl, and --NR.sub.fR.sub.g,
wherein R.sub.f and R.sub.g may be the same or different and are
chosen from hydrogen, (C.sub.1-C.sub.6)alkyl, and
(C.sub.3-C.sub.6)cycloalkyl.
[0034] 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.
[0035] A specific value for R.sub.1 is nitro or
NR.sub.aR.sub.b.
[0036] A specific value for R.sub.1 is nitro.
[0037] A specific value for R.sub.1 is NR.sub.aR.sub.b.
[0038] A specific value for R.sub.2 is hydrogen, or OR.sub.d,
wherein each R.sub.d is hydrogen or (C.sub.1-C.sub.6)alkyl.
[0039] A specific value for R.sub.2 is hydrogen.
[0040] A specific value for R.sub.2 is nitro or
NR.sub.aR.sub.b.
[0041] A specific value for R.sub.2 is nitro.
[0042] A specific value for R.sub.2 is NR.sub.aR.sub.b.
[0043] A specific value for R.sub.1 is hydrogen, or OR.sub.d,
wherein each R.sub.d is hydrogen or (C.sub.1-C.sub.6)alkyl.
[0044] A specific value for R.sub.1 is hydrogen.
[0045] A specific value for R.sub.3 is nitro or
NR.sub.aR.sub.b.
[0046] A specific value for R.sub.3 is nitro.
[0047] A specific value for R.sub.3 is NR.sub.aR.sub.b.
[0048] A specific value for R.sub.4 is hydrogen, or OR.sub.d,
wherein each R.sub.d is hydrogen or (C.sub.1-C.sub.6)alkyl.
[0049] A specific value for R.sub.4 is hydrogen.
[0050] A specific value for R.sub.4 is nitro or
NR.sub.aR.sub.b.
[0051] A specific value for R.sub.4 is nitro.
[0052] A specific value for R.sub.4 is NR.sub.aR.sub.b.
[0053] A specific value for R.sub.3 is hydrogen, or OR.sub.d,
wherein each R.sub.d is hydrogen or (C.sub.1-C.sub.6)alkyl.
[0054] A specific value for R.sub.3 is hydrogen.
[0055] A specific compound is a compound wherein R.sub.3 and
R.sub.4 taken together are methylenedioxy, 1,2-ethylenedioxy, or
1,3-propylenedioxy.
[0056] A specific compound is a compound wherein R.sub.3 and
R.sub.4 taken together are methylenedioxy.
[0057] A specific compound is a compound wherein R.sub.1 and
R.sub.2 taken together are methylenedioxy, 1,2-ethylenedioxy, or
1,3-propylenedioxy.
[0058] A specific compound is a compound wherein R.sub.1 and
R.sub.2 taken together are methylenedioxy.
[0059] A specific compound is a compound wherein R.sub.1 and
R.sub.2 are each independently OR.sub.d, wherein each R.sub.d is
hydrogen or (C.sub.1-C.sub.6)alkyl.
[0060] A specific compound is a compound wherein R.sub.1 and
R.sub.2 are each methoxy.
[0061] A specific compound is a compound wherein R.sub.3 and
R.sub.4 are each independently OR.sub.d, wherein each R.sub.d is
hydrogen or (C.sub.1-C.sub.6)alkyl.
[0062] A specific compound is a compound wherein R.sub.3 and
R.sub.4 are each methoxy. 5 A specific value for R.sub.5 is
(C.sub.1-C.sub.6)alkyl substituted with one or more hydroxy
groups.
[0063] Another specific value for R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one hydroxy group.
[0064] Another specific value for R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one or more mercapto groups.
[0065] Another specific value for R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one mercapto group.
[0066] Another specific value for R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one or more carboxy groups.
[0067] Another specific value for R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one carboxy group.
[0068] Another specific value for R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one or more NR.sub.fR.sub.ggroups.
[0069] Another specific value for R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one NR.sub.fR.sub.ggroup.
[0070] Another specific value for R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one or more NH.sub.2 groups.
[0071] Another specific value for R.sub.5 is a
(C.sub.1-C.sub.6)alkyl substituted with one NH.sub.2 group.
[0072] Another specific value for R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one or more N(CH.sub.3).sub.2 groups.
[0073] Another specific value for R.sub.5 is a
(C.sub.1-C.sub.6)alkyl substituted with one N(CH.sub.3).sub.2
group.
[0074] Another specific value for R.sub.5 is (C.sub.1-C.sub.6)alkyl
substituted with one or more N(CH.sub.2CH.sub.3).sub.2 groups.
[0075] Another specific value for R.sub.5 is a
(C.sub.1-C.sub.6)alkyl substituted with one
N(CH.sub.2CH.sub.3).sub.2 group.
[0076] Another specific value for R.sub.5 is a
(C.sub.1-C.sub.6)alkyl substituted with one or more
(C.sub.1-C.sub.6)alkoxycarbonyl (e.g. -CO.sub.2Me), cyano, halo,
hydroxy, mercapto, oxo (=O), carboxy (COOH), nitro, pyrrolidinyl,
piperidinyl, imidazolidinyl, imidazolinyl, piperazinyl,
morpholinyl, thiomorpholinyl, or -NR.sub.fR.sub.ggroups, wherein
R.sub.f and R.sub.g may be the same or different and are chosen
from hydrogen, (C.sub.1-C.sub.6)alkyl, and
(C.sub.3-C.sub.6)cycloalkyl.
[0077] Another specific value for R.sub.5 is a
(C.sub.2-C.sub.4)alkyl substituted with one or two groups selected
from hydroxy, mercapto, carboxy, amino, methylamino, ethylamino,
dimethylamino, and diethylamino.
[0078] Another specific value for R.sub.5 is 2-hydroxyethyl.
[0079] Another specific value for R.sub.5 is 3-hydroxypropyl.
[0080] Another specific value for R.sub.5 is 2-hydroxypropyl.
[0081] Another specific value for R.sub.5 is
-CH2CH2-NR.sub.fR.sub.gor
-CH.sub.2CH.sub.2CH.sub.2-NR.sub.fR.sub.gwherein R.sub.f and
R.sub.f are each independently hydrogen or
(C.sub.1-C.sub.6)alkyl.
[0082] Another specific value for R.sub.5 is
-CH.sub.2CH2-NR.sub.fR.sub.gor
-CH.sub.2CH.sub.2CH.sub.2-NR.sub.fR.sub.gwherein R.sub.f and
R.sub.g are each independently methyl or ethyl.
[0083] A specific compound is a compound wherein R.sub.1 is
hydrogen; R.sub.2 is nitro; and R.sub.3 and R.sub.4 taken together
are methylenedioxy.
[0084] A specific compound is a compound wherein R.sub.1 is nitro;
R.sub.2 is hydrogen; and R.sub.3 and R.sub.4 taken together are
methylenedioxy.
[0085] A specific compound is a compound wherein R.sub.1 and
R.sub.2 are each methoxy; R.sub.3 is nitro; and R.sub.4 is
hydrogen.
[0086] A specific compound is a compound wherein R.sub.1 and
R.sub.2 are each methoxy; R.sub.3 is hydrogen; and R.sub.4 is
nitro.
[0087] A specific compound is a compound wherein R.sub.1 is
hydrogen; R.sub.2 is nitro; and R.sub.3 and R.sub.4 are each
methoxy.
[0088] A specific compound is a compound wherein R.sub.1 is nitro;
R.sub.2 is hydrogen; and R.sub.3 and R.sub.4 are each methoxy.
[0089] A specific compound is a compound wherein R.sub.1 and
R.sub.2 taken together are methylenedioxy; R.sub.3 is nitro; and
R.sub.4 is hydrogen.
[0090] A specific compound is a compound wherein R.sub.1 and
R.sub.2 taken together are methylenedioxy; R.sub.3 is hydrogen; and
R.sub.4 is nitro.
[0091] A specific compound is a compound wherein the bond marked"a"
is a single bond (i.e. a compound of formula (III).
[0092] A specific compound is a compound wherein the bond marked"a"
is a single bond and the ring juncture at this bond is cis.
[0093] A specific compound is a compound wherein the bond marked"a"
is a single bond and the ring juncture at this bond is trans.
[0094] A specific compound is a compound wherein the bond marked"a"
is a double bond (i.e. a compound of formula (II).
[0095] A specific compound is any one of compounds 4a, 4b, 8a, and
8b wherein R.sub.5 is 2-(N,N-dimethylamino)ethyl,
2-(N,N-dimethylamino)propyl, 2-(N,N -diethylamino)ethyl, or 2-(N,N
-diethylamino)propyl; or a pharmaceutically acceptable salt
thereof.
[0096] A specific compound is any one of compounds 4a, 4b, 9a, and
9b wherein R.sub.5 is 2-(N,N-dimethylamino)ethyl,
2-(NdN-dimethylamino)propyl, 2-(N,N -diethylamino)ethyl, or 2-(N,N
-diethylamino)propyl; or a pharmaceutically acceptable salt
thereof.
[0097] A specific compound is any one of compounds 10a and 10b
wherein R.sub.5 is 2-(NN-dimethylamino)ethyl,
2-(N,N-dimethylamino)propyl, 2-(N,N -diethylamino)ethyl, or
2-(N,N-diethylamino)propyl; or a pharmaceutically acceptable salt
thereof.
[0098] A specific compound is any one of compounds 14a, 14b, 16a,
and 16b wherein n is 1, 2, or 3; and Y is dimethylamino or
diethylamino; or a pharmaceutically acceptable salt thereof.
[0099] A specific compound is any one of compounds 20a, 20b, 22a,
and 22b wherein n is 1, 2, or 3; and Y is dimethylamino or
diethylamino; or a pharmaceutically acceptable salt thereof.
[0100] A compound of formula I can be prepared using procedures
similar to those described in International Patent Application
Publication Number 00/21537 or as illustrated in FIG. 1. Methods
for the preparation of 1 have been described (see J. Chem. Soc.,
1955, 2675-2685). Reaction of the appropriate benzaldehyde with a
primary amine (R.sub.5-NH.sub.2) provides the Shiff base
intermediates (2a and 2b). Reaction of 2a or 2b with homophthalic
anhydride 1 provides the
4-carboxy-N-substituted-3,4-dihydro-3-phenyl-2H-isoquinolin-1-ones
(3a and 3b). Treatment with thionyl chloride provides the compounds
of formula (I) (4a and 4b). Alternate treatment with Eaton's
reagent (10% P.sub.2O.sub.5 in methanesulfonic acid) instead of
thionyl chloride provides the compounds of formula (I) (5a and
5b).
[0101] Compounds of formula I can also be prepared as illustrated
in FIG. 2. Reaction of 3,4-methylenedioxybenzaldehyde with a
primary amine (R.sub.5-NH.sub.2) provides the Shiff base
intermediate 7. Reaction 6a or 6b with compound 7 provides
cis-4-carboxy-3,4-dihydro-N-substituted-3-(3',4'-methylenedioxypheny)-1(2-
H)isoquinolones 8a and 8b. Treatment with thionyl chloride provides
compounds 9a and 9b. Alternate treatment with Eaton's reagent (10%
P.sub.2O.sub.5 in methanesulfonic acid) instead of thionyl chloride
provides the compounds of formula (I) (l0a and 10b).
[0102] Compounds of formula I can also be prepared as illustrated
in FIG. 3. Reaction of compound 1 with compound 11a or 11b (wherein
X is, for example, hydroxy, protected hydroxy, halo, or cyano)
provides compounds 12a or 12b, which can be cyclized to provide
compounds 13a or 13b, which are compounds of formula (I).
Subsequent conversion, for example of a compound wherein X is halo,
provides additional compounds of formula (I) (compounds 14a or 14b)
wherein Y is NR.sub.aR.sub.bor CH.sub.2NR.sub.aR.sub.b. Alternate
treatment of compounds 12a or 12b with Eaton's reagent (10%
P.sub.2O.sub.5 in methanesulfonic acid) instead of thionyl chloride
provides the compounds of formula (I) (15a and 15b). Subsequent
conversion, for example of a compound wherein X is halo, provides
additional compounds of formula (I) (compounds 16a or 16b) wherein
Y is NR.sub.aR.sub.bor CH.sub.2NR.sub.aR.sub.b.
[0103] Compounds of formula I can also be prepared as illustrated
in FIG. 4. Reaction of compound 5a or 5b with compound 17 (wherein
X is, for example, hydroxy, protected hydroxy, halo, or cyano)
provides compounds 18a or 18b, which can be cyclized to provide
compounds 19a or 19b, which are compounds of formula (I).
Subsequent conversion, for example of a compound wherein X is halo,
provides additional compounds of formula (I) (compounds 20a or 20b)
wherein Y is NR.sub.aR.sub.bor CH2R.sub.zR.sub.b Alternate
treatment of compounds 18a or 18b with Eaton's reagent (10%
P.sub.2O.sub.5 in methanesulfonic acid) instead of thionyl chloride
provides the compounds of formula (I) (21a and 21b). Subsequent
conversion, for example of a compound wherein X is halo, provides
additional compounds of formula (I) (compounds 22a or 22b) wherein
Y is NR.sub.aR.sub.bor CH2NR.sub.aR.sub.b.
[0104] Compounds wherein R.sub.1-R.sub.4 are amino can be prepared
from the corresponding compounds wherein R.sub.1-R.sub.4 are nitro
by reduction of the nitro group using procedures that are known,
such as, for example, with R.sub.a iny nickel and hydrazine.
Additionally, standard methods can be used to substitute the
resulting aryl amines to provide additional compounds of the
invention.
[0105] The starting materials employed in the synthetic methods
described herein are commercially available, have been reported in
the scientific literature, or can be prepared from readily
available starting materials using procedures known in the field.
It may be desirable to optionally use a protecting group during all
or portions of the above described synthetic procedures. Such
protecting groups and methods for their introduction and removal
are well known in the art. See Greene, T.W.; Wutz,
P.G.M."Protecting Groups In Organic Synthesis" second edition,
1991, New York, John Wiley & Sons, Inc.
[0106] 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 inhibition activity or cytotoxic activity using the
standard tests described herein, or using other similar tests which
are well known in the art.
[0107] 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.
[0108] 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.
[0109] 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, that is, orally or parenterally, by
intravenous, intramuscular, topical or subcutaneous routes.
[0110] Thus, the present compounds may be systemically
administered, for example, 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.
[0111] 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.
[0112] 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.
[0113] 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 must 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 antifuingal agents,
for example, parabens, chlorobutanol, phenol, sorbic acid,
thimerosal, and the like. In many cases, it will be preferable to
include isotonic agents, for example, sugars, buffers or sodium
chloride.
[0114] 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.
[0115] 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.
[0116] 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.
[0117] 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.
[0118] 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.
[0119] 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).
[0120] 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.
[0121] 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-%.
[0122] 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.
[0123] In general, however, a suitable dose will be in the range of
from about 0.5 to about 100 mg/kg per day, e.g., from about 1 to
about 60-mg/kg of body weight per day or about 2 to 50 mg/kg per
day.
[0124] The compound may conveniently be administered in unit dosage
form; for example, containing 5 to 1,000 mg, conveniently 10 to 750
mg, most conveniently, 50 to 500 mg of active ingredient per unit
dosage form.
[0125] 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.
[0126] The ability of a compound of the invention to effect
topoisomerase I or II 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.
Test A. Topoisomerase-mediated DNA cleavage assays.
[0127] Human topoisomerase I was expressed in E. Coli and isolated
as a recombinant fusion protein using a T7 expression system as
described previously (Gatto, B., Sanders, M. M., Yu, C., Wu, H.-Y.,
Makhey, D., LaVoie, E. J., and Liu, L. F. (1996) Cancer Res. 56,
2795-2800). Recombinant human topoisomerase IIc was isolated and
purified as previously described (Wasserman, R.sub.1 A. Austin,
C.A., Fisher, L.M.; Wang, J. C., Cancer Res., 1993, 53, 3591;
Halligan, B. D.; Edwards, K. A.; Liu, L. F. J. Biol. Chem 1985,
260, 2475). Plasmid YepG was also purified by the alkali lysis
method followed by phenol deproteination and CsCI/ethidium
isopycnic centrifugation method as described. The end-labeling of
the plasmid was accomplished by digestion with a restriction enzyme
followed by end-filling with Klenow polymerase as previously
described (Maniatis, T.; Fritsch, E. F.; Sambrook, J. Molecular
Cloning, a Laboratory Manual; Cold Spring Harbor Laboratory: Cold
Spring Harbor, NY 1982; pp 149-185.). The cleavage assays were
performed as previously reported (Gatto, B., Sanders, M. M., Yu,
C., Wu, H.-Y., Makhey, D., LaVoie, E. J., and Liu, L. F. (1996)
Cancer Res. 56, 2795-2800; Tewey, K. M., Rowe, T. C., Yang, L.,
Hallogan, B. C., and Liu, L. F. (1984) Science 226, 466-468; Li
T-K., Chen AY, Yu C.sub.1Mao Y, Wang H, Liu LF. (1999) Genes Dev
13(12):1553-60; Wang, H.; Mao, Y.; Chen, A.Y.; Zhou, N.; and
LaVoie, E.J.; Liu, L.F. Biochemistry, 2001, 40, 3316). The drug and
the DNA in presence of topoisomerase I was incubated for 30 minutes
at 37 .degree. C. After development of the gels, typically 24-hour
exposure was used to obtain autoradiograms outlining the extent of
DNA fragmentation. Topoisomerase I-mediated DNA cleavage values are
reported as REC.sub.1Relative Effective Concentration, i.e.
concentrations relative to topotecan, whose value is arbitrarily
assumed as 1.0, that are able to produce the same cleavage on the
plasmid DNA in the presence of human topoisomerase I. Topoisomerase
II-mediated DNA cleavage values are reported as REC.sub.1Relative
Effective Concentration, potency was based upon the relative amount
of drug needed to induce approximately 10% DNA fragmentation, i.e.
concentrations relative to VM-26, whose value is arbitrarily
assumed as 1.0, that are able to produce the same cleavage on the
plasmid DNA in the presence of human topoisomerase II.
[0128] 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. Test
B. Inhibition of Cell Growth: MTT-microtiter plate tetrazolinium
cytotoxicity assay (RPMI 8402, CPT-K5, U937, U937/CR Cells) The
cytotoxicity is 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 (Anchi Cancer
Research Institute, Nagoya, Japan), see Andoh, T.; Okada, K, Adv.
in Pharmacology 1994, 29B, 93. Human U-937 myeloid leukemia cells
and U-937/CR cells were described by Rubin et al., J. Biol. Chem.,
1994, 269, 2433-2439. The cytotoxicity assay is performed by using
96-well microtiter plates using 2000 cells/well, in 200 mL of
growth medium. Cells are 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 (100U/mL), and streptomycin (0.1
mg/mL). For determination of IC.sub.50, cells are exposed
continuously for 3-4 days to varying concentrations of drug, and
MTT assays were performed at the end of the fourth day. Each assay
is performed with a control that did not contain any drug. All
assays are performed at least twice in 6 replicate wells. All
assays are performed under the direction of Dr. L. F. Liu,
Department of Pharmacology, The University of Medicine and
Dentistry of New Jersey, Robert Wood Johnson Medical School,
Piscataway, New Jersey. Data for representative compound B is
provided in Table 1. TABLE-US-00001 TABLE 1 Cell Lines RPMI8402
CPT-K5 KB3-1 KBV-1 KBH5.0 HL60 HL60/MX2 0.003* 2.4 0.004 0.004
0.004 0.003 0.003 *Cytotoxicity (.mu.M IC.sub.50 values)
[0129] The compounds of the invention can function as cytotoxic
agents against tumor cell lines, including multi-drug resistant
tumor cell lines. Thus, the compounds are useful to treat cancer
and can be used to treat tumors that are resistant to other
specific chemotherapeutic agents.
[0130] Topoisomerase inhibitors are also known to possess
antibacterial, antifungal, antiprotozoal, antihelmetic,
antipsoriatic, and antiviral activity. Accordingly, the
topoisomerase inhibitors of the. invention may also be useful as
antibacterial, antifungal, antiprotozoal, antihelmetic,
antipsoriatic, or antiviral agents. In particular, compounds of the
invention that demonstrate little or no activity as mammalian
topoisomerase I poisons, because of the possibility of similar
molecular mechanism of action, could be highly active and selective
antibacterial, antifungal, antiprotozoal, antihelmetic,
antipsoriatic, or antiviral agents. Thus, certain compounds of the
invention may be particularly useful as systemic antibacterial,
antifungal, antiprotozoal, antihelmetic, antipsoriatic, or
antiviral agents in mammals. The invention also provides the use of
a compound of the invention for the manufacture of a medicament
useful for producing an antibacterial, antifungal, antiprotozoal,
antihelmetic, antipsoriatic, or antiviral effect in a mammal.
[0131] As used herein, the term"solid mammalian tumors" include
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.
[0132] The invention will now be illustrated by the following
non-limiting Example.
EXAMPLES
Example 1
6-[3-(N,N-dimethylamino)propyll-3-Nitroindeno[1,2-c]-indenoisoquinolin-5,1-
1-dione (Compound B, Fugure 5).
[0133] ##STR3##
[0134] The title compound was prepared as illustrated in FIG. 5. To
a solution of Compound A (300 mg, 0.66 mmol) in chloroform (50 mL)
and methanol (50 mL) was added a 2.0 M solution of dimethylamine in
methanol (6 mL), and the resulting mixture was heated in a steel
bomb to 140.degree. C., and maintained at this temperature with
stirring for 48 h. The reaction mixture was cooled to room
temperature and the solvent was removed under vacuum. To the
residue was added water, and the resulting suspension was basified
(10% NaOH), extracted with CHCI.sub.3, and chromatographed on
silica eluting with 95:5 chloroform-methanol, to provide 60 mg of
the title compound as a red solid, in 21% yield; mp 191-192.degree.
C.; .sup.1H NMR (CDCl.sub.3) 2.04 (m, 2H), 2.38 (s,6H), 2.58 (m,
2H), 4.57 (t, 2H, J=7.9), 6.16 (s, 2H), 7.18 (s, 1H), 7.72 (s, 1H),
8.45 (dd, 1H, J=9.1, J=2.3), 8.76 (d, 1H, J=9.1), 9.17 (d, 1H,
J=2.3); IR (KBr) 1697, 1674, 1499, 1337; HRMS calcd for
C.sub.22H.sub.19N.sub.3O.sub.6H: 422.1352; found: 422.1357.
[0135] The intermediate Compound A was prepared as follows.
[0136] a. Compound A. 4-Nitrohomophthalic anhydride (4.14 g, 20.0
mmol, see Whitmore, W.F., et al., J. Am. Chem. Soc., 1944, 66,
1237-1240) was added to a solution of
3,4-methylenedioxybenzylidene-(3-bromo-l-propylamine) (5.4 g, 20.0
nunol, see Cushman, M., et al., J. Med. Chem., 2000, 43, 3688-3698)
in chloroform (200 mL), and the resulting mixture was stirred at
room temperature overnight. The material that precipitated during
the course of the reaction was isolated by filtration and was
washed with chloroform. Drying yielded 6.3 g of material containing
of a mixture of isomers. The mixture not characterized or purified
further at this stage. 3.0 g (6.3 mmol) of this material was added
to 12 mL of thionyl chloride, and the resulting mixture was stirred
at room temperature overnight. Benzene was added to the red
solution and it was concentrated under reduced pressure. Chloroform
was added to the residue and the mixture was filtered through a
short column of silica, providing 350 mg of a dark brown solid, in
8% yield; mp 281-282.degree. C.; .sub.1H NMR (CDCl.sub.3) 2.19 (m,
2H), 3.69 (t, 2H, J=6.0), 4.66 (t, 2H, J=8.1), 6.18 (s, 2H), 7.21
(s, 1H), 7.49 (s, IH), 8.49 (dd, 1H, J=9.2, J=2.6), 8.79 (d, IH,
J=9.2), 9.18 (d, 1H, J=2.6); IR (KBr) 1698, 1658, 1504, 1333; HRMS
calcd for C.sub.20Hl.sub.13N.sub.2O.sub.6BrH: 457.0037; found:
457.0035.
Example 2
[0137] The following illustrate representative pharmaceutical
dosage forms, containing a compound of formula I (.degree. Compound
X'), for therapeutic or prophylactic use in humans. TABLE-US-00002
(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 1.0 N 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 01 N 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 Trichloromonofluoromethane
5,000.0 Dichlorodifluoromethane 10,000.0 Dichlorotetrafluoroethane
5,000.0
[0138] The above formulations may be obtained by conventional
procedures well known in the pharmaceutical art.
[0139] 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.
* * * * *