U.S. patent application number 15/241240 was filed with the patent office on 2016-12-08 for compositions and methods for treatment of cancer.
The applicant listed for this patent is ArQule, Inc.. Invention is credited to Mark A. Ashwell, Jason Hill, Chiang J. Li, Magdi Moussa, Neru Munshi.
Application Number | 20160355515 15/241240 |
Document ID | / |
Family ID | 36579999 |
Filed Date | 2016-12-08 |
United States Patent
Application |
20160355515 |
Kind Code |
A1 |
Li; Chiang J. ; et
al. |
December 8, 2016 |
COMPOSITIONS AND METHODS FOR TREATMENT OF CANCER
Abstract
The present invention relates to
pyrroloquinolinyl-pyrrolidine-2,5-dione compounds in combination
with chemotherapeutic agents. The present invention provides
methods of treating a cell proliferative disorder, such as a
cancer, by administering to a subject in need thereof a
therapeutically effective amount of a
pyrroloquinolinyl-pyrrole-2,5-dione compound or
pyrroloquinolinyl-pyrrolidine-2,5-dione compound of the present
invention and also administering a effective amount of a
chemotherapeutic agent.
Inventors: |
Li; Chiang J.; (Cambridge,
MA) ; Ashwell; Mark A.; (Carlisle, MA) ; Hill;
Jason; (Auburndale, MA) ; Moussa; Magdi;
(Malden, MA) ; Munshi; Neru; (Burlington,
MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ArQule, Inc. |
Burlington |
MA |
US |
|
|
Family ID: |
36579999 |
Appl. No.: |
15/241240 |
Filed: |
August 19, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14215852 |
Mar 17, 2014 |
9447088 |
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15241240 |
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13761712 |
Feb 7, 2013 |
8754078 |
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14215852 |
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12722394 |
Mar 11, 2010 |
8377927 |
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13761712 |
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11350335 |
Feb 9, 2006 |
7713969 |
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12722394 |
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60650951 |
Feb 9, 2005 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 471/06 20130101;
A61K 31/513 20130101; A61K 38/08 20130101; A61K 31/683 20130101;
A61K 31/7048 20130101; A61P 35/04 20180101; A61P 35/02 20180101;
A61K 38/12 20130101; A61K 31/337 20130101; A61K 31/5517 20130101;
A61K 31/4535 20130101; C07D 455/00 20130101; C07F 9/6561 20130101;
A61K 31/138 20130101; C07D 487/06 20130101; C07K 2/00 20130101;
A61P 35/00 20180101; A61K 31/4745 20130101; C07F 9/65583 20130101;
A61K 31/675 20130101; A61K 31/5377 20130101; A61K 31/7068 20130101;
A61K 31/704 20130101; A61K 31/664 20130101 |
International
Class: |
C07D 471/06 20060101
C07D471/06; A61K 31/5377 20060101 A61K031/5377; A61K 31/5517
20060101 A61K031/5517; A61K 31/683 20060101 A61K031/683; C07D
487/06 20060101 C07D487/06; A61K 31/4745 20060101 A61K031/4745;
C07F 9/6561 20060101 C07F009/6561 |
Claims
1. A pyrroloquinolinyl-pyrrolidine-2,5-dione compound of formula
IVa, IVb, Va, or Vb, or a pharmaceutically acceptable salt thereof:
##STR00036## wherein: R1, R2 and R3 are independently selected from
the group consisting of hydrogen, F, Cl, Br, I, --NR5R6,
--(C.sub.1-C.sub.6) alkyl, --(C.sub.1-C.sub.6) substituted alkyl,
--(C.sub.3-C.sub.9) cycloalkyl, --(C.sub.3-C.sub.9) substituted
cycloalkyl, --O--(C.sub.1-C.sub.6) alkyl, --O--(C.sub.1-C.sub.6)
substituted alkyl, --O--(C.sub.3-C.sub.9) cycloalkyl,
--O--(C.sub.3-C.sub.9) substituted cycloalkyl, aryl, heteroaryl,
and heterocyclyl; R4 is selected from the group consisting of
hydrogen, --(C.sub.1-C.sub.6) alkyl, and --CH.sub.2R7; R5 and R6
are independently selected from the group consisting of hydrogen
and --(C.sub.1-C.sub.6) alkyl; R7 is selected from the group
consisting of --O--P(.dbd.O)(OH).sub.2,
--O--P(.dbd.O)(--OH)(--O--(C.sub.1-C.sub.6) alkyl),
--O--P(.dbd.O)(--O--(C.sub.1-C.sub.6) alkyl).sub.2,
--O--P(.dbd.O)(--OH)(--O--(CH.sub.2)-phenyl),
--O--P(.dbd.O)(--O--(CH.sub.2)-phenyl).sub.2, a carboxylic acid
group, an amino carboxylic acid group, and a peptide; Q is selected
from the group consisting of aryl, heteroaryl, --O-aryl, --S-aryl,
--O-- heteroaryl, and --S-heteroaryl; X is selected from the group
consisting of --(CH.sub.2)--, S, --(NR8)-, and O; R8 is
independently selected from the group consisting of hydrogen,
--(C.sub.1-C.sub.6) alkyl, --(C.sub.1-C.sub.6) substituted alkyl,
--(C.sub.3-C.sub.9) cycloalkyl, --(C.sub.3-C.sub.9) substituted
cycloalkyl, and --O--(C.sub.1-C.sub.6) alkyl,
--C(.dbd.O)--O--(C.sub.1-C.sub.6) alkyl and
--C(.dbd.O)--O--(C.sub.1-C.sub.6) substituted alkyl; Y is selected
from the group consisting of --(CH.sub.2)-- and a bond; wherein
said aryl, heteroaryl, --O-aryl, --S-aryl, --O-heteroaryl, and
--S-heteroaryl may be substituted with one or more substituents
independently selected from the group consisting of F, Cl, Br, I,
--NR5R6, --(C.sub.1-C.sub.6) alkyl, --(C.sub.1-C.sub.6) substituted
alkyl, --(C.sub.3-C.sub.9) cycloalkyl, --(C.sub.3-C.sub.9)
substituted cycloalkyl, --O--(C.sub.1-C.sub.6) alkyl,
--O--(C.sub.1-C.sub.6) substituted alkyl, --O--(C.sub.3-C.sub.9)
cycloalkyl, --O--(C.sub.3-C.sub.9) substituted cycloalkyl, -aryl,
-aryl-(C.sub.1-C.sub.6) alkyl, -aryl-O--(C.sub.1-C.sub.6) alkyl,
--O-aryl, --O--(C.sub.1-C.sub.4) alkyl-aryl, heteroaryl,
heterocyclyl, --O--(C.sub.1-C.sub.4) alkyl-heterocycle, and
--(S(.dbd.O).sub.2)--(C.sub.1-C.sub.6) alkyl; and m is 1 or 2.
2. The compound of claim 1, wherein Q is an indolyl group or an
indolyl group substituted with one or more substituents
independently selected from the group consisting of F, Cl, Br, I,
--(C.sub.1-C.sub.6) alkyl, --(C.sub.1-C.sub.6) fluoro-substituted
alkyl, --(C.sub.3-C.sub.9) cycloalkyl, --(C.sub.3-C.sub.9)
fluoro-substituted cycloalkyl, --O--(C.sub.1-C.sub.6) alkyl,
--O--(C.sub.1-C.sub.6) fluoro-substituted alkyl,
--O--(C.sub.3-C.sub.9) cycloalkyl, --O--(C.sub.3-C.sub.9)
fluoro-substituted cycloalkyl, -aryl, --O-aryl,
--O--(C.sub.1-C.sub.4) alkyl-aryl, --O--(C.sub.1-C.sub.4)
alkyl-heterocycle, and --S(.dbd.O).sub.2--(C.sub.1-C.sub.6)
alkyl.
3. The compound of claim 1, wherein R4 is --CH.sub.2R7, and R7 is
--O--P(.dbd.O)(OH).sub.2,
--O--P(.dbd.O)(--OH)(--O--(C.sub.1-C.sub.6) alkyl),
--O--P(.dbd.O)(--O--(C.sub.1-C.sub.6) alkyl).sub.2, a carboxylic
acid group, an amino carboxylic acid group, or a peptide.
4. The compound of claim 3, wherein R7 is --O--P(.dbd.O)(OH).sub.2,
--O--P(.dbd.O)(--OH)(--O--(C.sub.1-C.sub.6) alkyl), or
--O--P(.dbd.O)(--O--C.sub.1-C.sub.6)alkyl).sub.2.
5. The compound of claim 3, wherein R7 is a carboxylic acid group,
an amino carboxylic acid group, or a peptide.
6. The compound of claim 5, wherein R7 is alanine, arginine,
asparagine, aspartic acid, cysteine, glutamine, glutamic acid,
glycine, histidine, isoleucine, leucine, lysine, methionine,
phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or
valine.
7. The compound of claim 6, wherein R7 is L-alanine, L-arginine,
L-asparagine, L-aspartic acid, L-cysteine, L-glutamine, L-glutamic
acid, L-glycine, L-histidine, L-isoleucine, L-leucine, L-lysine,
L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine,
L-tryptophan, L-tyrosine, or L-valine.
8. The compound of claim 5, wherein R7 is a peptide.
9. The compound of claim 8, wherein said peptide is comprised of
two or more imino or amino acids selected from the group consisting
of: L-alanine, L-arginine, L-asparagine, L-aspartic acid,
L-cysteine, L-glutamine, L-glutamic acid, L-glycine, L-histidine,
L-isoleucine, L-leucine, L-lysine, L-methionine, L-phenylalanine,
L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, and
L-valine.
10. The compound of claim 1, wherein R1, R2 and R3 are each
hydrogen.
11. The compound of claim 1, wherein R4 is hydrogen.
12. A method of treating a cell proliferative disorder, comprising
administering to a subject in need thereof a therapeutically
effective amount of a compound of claim 1 or a pharmaceutically
acceptable salt thereof, wherein the cell proliferative disorder is
selected from lung cancer, colon cancer, breast cancer, pancreatic
cancer, prostate cancer, chronic myelogenous leukemia, melanoma,
and ovarian cancer.
13. A pharmaceutical composition comprising a compound of claim 1
or a pharmaceutically acceptable salt thereof together with one or
more pharmaceutically acceptable carriers or excipients.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. Ser. No.
14/215,852, filed Mar. 17, 2014, allowed, which is a divisional of
U.S. Ser. No. 13/761,712, filed Feb. 7, 2013, now U.S. Pat. No.
8,754,078, which is a divisional of U.S. Ser. No. 12/722,394, filed
Mar. 11, 2010, now U.S. Pat. No. 8,377,927, which is a divisional
of U.S. Ser. No. 11/350,335, filed Feb. 9, 2006, now U.S. Pat. No.
7,713,969, which claims priority to and the benefit of U.S. Ser.
No. 60/650,951, filed Feb. 9, 2005, the entire contents of each of
which are incorporated herein by reference.
BACKGROUND OF THE INVENTION
[0002] Cancer is the second leading cause of death in the United
States, exceeded only by heart disease. (Cancer Facts and Figures
2004, American Cancer Society, Inc.) Despite recent advances in
cancer diagnosis and treatment, surgery and radiotherapy may be
curative if a cancer is found early, but current drug therapies for
metastatic disease are mostly palliative and seldom offer a
long-term cure. Even with new chemotherapies entering the market,
the need continues for new drugs effective in monotherapy or in
combination with existing agents as first line therapy, and as
second and third line therapies in treatment of resistant
tumors.
[0003] Cancer cells are by definition heterogeneous. For example,
within a single tissue or cell type, multiple mutational
`mechanisms` may lead to the development of cancer. As such,
heterogeneity frequently exists between cancer cells taken from
tumors of the same tissue and same type that have originated in
different individuals. Frequently observed mutational `mechanisms`
associated with some cancers may differ between one tissue type and
another (e.g., frequently observed mutational `mechanisms` leading
to colon cancer may differ from frequently observed `mechanisms`
leading to leukemias). It is therefore often difficult to predict
whether a particular cancer will respond to a particular
chemotherapeutic agent. (Cancer Medicine, 5th Edition, Bast et al.
eds., B.C. Decker Inc., Hamilton, Ontario)
[0004] Breast cancer is the most frequently diagnosed non-skin
cancer in women, and ranks second among cancer deaths in women,
after lung cancer. (Cancer Facts and Figures 2004, American Cancer
Society, Inc.) Current treatment options for breast cancer include
surgery, radiotherapy, and chemotherapy/hormone therapy with agents
such as tamoxifen, aromatase inhibitors, HERCEPTIN.RTM.
(trastuzumab), TAXOL.RTM. (paclitaxel), cyclophosphamide,
methotrexate, doxorubicin (adriamycin), and 5-fluoruracil. Despite
improvements in cancer diagnostics and therapeutics, breast cancer
incidence rates have continued to increase since 1980. In 2004,
about 215,000 new cases of breast cancer are expected in women, and
about 1,450 new cases of breast cancer are expected in men. Id.
Accordingly, new compounds and methods for treating breast cancer
are needed.
[0005] Components of cellular signal transduction pathways that
regulate the growth and differentiation of normal cells can, when
dysregulated, lead to the development of cellular proliferative
disorders and cancer. Mutations in cellular signaling proteins may
cause such proteins to become expressed or activated at
inappropriate levels or at inappropriate times during the cell
cycle, which in turn may lead to uncontrolled cellular growth or
changes in cell-cell attachment properties. For example,
dysregulation of receptor tyrosine kinases by mutation, gene
rearrangement, gene amplification, and overexpression of both
receptor and ligand has been implicated in the development and
progression of human cancers.
[0006] The c-Met receptor tyrosine kinase is the only known
high-affinity receptor for hepatocyte growth factor (HGF), also
known as scatter factor. Binding of HGF to the c-Met extracellular
ligand-binding domain results in receptor multimerization and
phosphorylation of multiple tyrosine residues in the intracellular
portion of c-Met. Activation of c-Met results in the binding and
phosphorylation of adaptor proteins such as Gab-1, Grb-2, She, and
c-Cbl, and subsequent activation of signal transducers such as
PI3K, PLC-.gamma., STATs, ERK1 and 2 and FAK. c-Met and HGF are
expressed in numerous tissues, and their expression is normally
confined predominantly to cells of epithelial and mesenchymal
origin, respectively. c-Met and HGF are dysregulated in human
cancers, and may contribute to dysregulation of cell growth, tumor
cell dissemination, and tumor invasion during disease progression
and metastasis. (See, e.g., Journal of Clinical Investigation 109:
863-867 (2002) and Cancer Cell pp 5-6 Jul. 2004) c-Met and HGF are
highly expressed relative to surrounding tissue in numerous
cancers, and their expression correlates with poor patient
prognosis. (See, e.g., Journal of Cellular Biochemistry 86: 665-677
(2002); Int. J. Cancer (Pred. Oncol.) 74: 301-309 (1997); Clinical
Cancer Research 9: 1480-1488 (2003); and Cancer Research 62:
589-596 (2002)). Without intending to be bound by theory, c-Met and
HGF may protect tumors against cell death induced by DNA damaging
agents, and as such may contribute to chemoresistance and
radioresistance of tumors. Without intending to be limited by any
theory, inhibitors of c-Met may be useful as therapeutic agents in
the treatment of proliferative disorders including breast cancer.
(See, e.g., Cancer and Metastasis Reviews 22: 309-325 (2003)).
[0007] The references cited herein are not admitted to be prior art
to the claimed invention.
SUMMARY OF THE INVENTION
[0008] The present invention provides for
pyrroloquinolinyl-pyrrolidine-2,5-dione compounds of formula IVa,
IVb, Va, or Vb, and methods of preparing the compounds of formula
IVa, IVb Va, and Vb,
##STR00001##
where:
[0009] R1, R2 and R3 are independently selected from the group
consisting of hydrogen, F, Cl, Br, I, --NR5R6, --(C.sub.1-C.sub.6)
alkyl, --(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl,
--O--(C.sub.1-C.sub.6) alkyl, --O--(C.sub.1-C.sub.6) substituted
alkyl, --O--(C.sub.3-C.sub.9) cycloalkyl, and
--O--(C.sub.3-C.sub.9) substituted cycloalkyl, aryl, heteroaryl,
heterocyclyl;
[0010] R4 is independently selected from the group consisting of
hydrogen, --(C.sub.1-C.sub.6) alkyl, --CH.sub.2R7;
[0011] R5, R6 are independently selected from the group consisting
of hydrogen, and --(C.sub.1-C.sub.6) alkyl:
[0012] R7 is independently selected from the group consisting of
--O--P(.dbd.O)(OH).sub.2,
--O--P(.dbd.O)(--OH)(--O--(C.sub.1-C.sub.6) alkyl),
--O--P(.dbd.O)(--O--(C.sub.1-C.sub.6) alkyl).sub.2,
--O--P(.dbd.O)(--OH)(--O--(CH.sub.2)-phenyl),
--O--P(.dbd.O)(--O--(CH.sub.2)-phenyl).sub.2, a carboxylic acid
group, an amino carboxylic acid group and a peptide;
[0013] Q is selected from the group consisting of aryl, heteroaryl,
--O-aryl, --S-aryl, --O-heteroaryl, and --S-heteroaryl;
[0014] X is selected from the group consisting of --(CH.sub.2)--,
--(NR8)-, S, and O;
[0015] R8 is independently selected from the group consisting of
hydrogen, --(C.sub.1-C.sub.6) alkyl, --(C.sub.1-C.sub.6)
substituted alkyl, --(C.sub.3-C.sub.9) cycloalkyl,
--(C.sub.3-C.sub.9) substituted cycloalkyl, and
--O--(C.sub.1-C.sub.6) alkyl, --C(.dbd.O)--O--(C.sub.1-C.sub.6)
alkyl and --C(.dbd.O)--O--(C.sub.1-C.sub.6) substituted alkyl;
[0016] Y is selected from the group consisting of --(CH.sub.2)-- or
a bond;
[0017] wherein said aryl, heteroaryl, --O-aryl, --S-aryl,
--O-heteroaryl, and --S-heteroaryl groups may be substituted with
one or more substituents independently selected from the group
consisting of F, Cl, Br, I, --NR5R6, --(C.sub.1-C.sub.6) alkyl,
--(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl,
--O--(C.sub.1-C.sub.6) alkyl, --O--(C.sub.1-C.sub.6) substituted
alkyl, --O--(C.sub.3-C.sub.9) cycloalkyl, --O--(C.sub.3-C.sub.9)
substituted cycloalkyl, -aryl, -aryl-(C.sub.1-C.sub.6) alkyl,
-aryl-O--(C.sub.1-C.sub.6) alkyl, --O-aryl, --O--(C.sub.1-C.sub.4)
alkyl-aryl, heteroaryl, heterocyclyl, --O--(C.sub.1-C.sub.4)
alkyl-heterocycle, and --(S(.dbd.O).sub.2)--(C.sub.1-C.sub.6)
alkyl; and
[0018] m is 1 or 2.
[0019] In an embodiment, R4 is --CH.sub.2R7, and R7 is
--O--P(.dbd.O)(OH).sub.2,
--O--P(.dbd.O)(--OH)(--O--(C.sub.1-C.sub.6) alkyl),
--O--P(.dbd.O)(--O--(C.sub.1-C.sub.6) alkyl).sub.2, a carboxylic
acid group, an amino carboxylic acid group or a peptide.
[0020] In an embodiment, X is selected from the group consisting of
--(NR8)-, S, and O.
[0021] In an embodiment, m is 2.
[0022] In a preferred embodiment, the
pyrroloquinolinyl-pyrrolidine-2,5-dione compound is selected from
the group consisting of (+)-cis-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione,
(-)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione,
(+)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione, and
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione. In a
further preferred embodiment, the compound is
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione.
[0023] The present invention also provides for
pyrroloquinolinyl-pyrrole-2,5-dione compounds of formula IIIa and
their synthesis.
##STR00002##
where:
[0024] R1, R2 and R3 are independently selected from the group
consisting of hydrogen, F, Cl, Br, I, --NR5R6, --(C.sub.1-C.sub.6)
alkyl, --(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl,
--O--(C.sub.1-C.sub.6) alkyl, --O--(C.sub.1-C.sub.6) substituted
alkyl, --O--(C.sub.3-C.sub.9) cycloalkyl, and
--O--(C.sub.3-C.sub.9) substituted cycloalkyl, aryl, heteroaryl,
heterocyclyl;
[0025] R4 is independently selected from the group consisting of
hydrogen, --(C.sub.1-C.sub.6) alkyl, --CH.sub.2R7;
[0026] R5, R6 are independently selected from the group consisting
of hydrogen, and --(C.sub.1-C.sub.6) alkyl;
[0027] R7 is independently selected from the group consisting of
--O--P(.dbd.O)(OH).sub.2,
--O--P(.dbd.O)(--OH)(--O--(C.sub.1-C.sub.6) alkyl),
--O--P(.dbd.O)(--O--(C.sub.1-C.sub.6) alkyl).sub.2,
--O--P(.dbd.O)(--OH)(--O--(CH.sub.2)-phenyl),
--O--P(.dbd.O)(--O--(CH.sub.2)-phenyl).sub.2, a carboxylic acid
group, an amino carboxylic acid group and a peptide;
[0028] Q is selected from the group consisting of aryl, heteroaryl,
--O-aryl, --S-aryl, --O-heteroaryl, and --S-heteroaryl, provided
that when R4 is hydrogen, (C.sub.3-C.sub.4) cycloalkyl, or
(C.sub.1-C.sub.4) alkyl, Q is not 3-indolyl or substituted
3-indolyl;
[0029] X is selected from the group consisting of --(CH.sub.2)--,
--(NR8)-, S, and O;
[0030] R8 is independently selected from the group consisting of
hydrogen, --(C.sub.1-C.sub.6) alkyl, --(C.sub.1-C.sub.6)
substituted alkyl, --(C.sub.3-C.sub.9) cycloalkyl,
--(C.sub.3-C.sub.9) substituted cycloalkyl, --O--(C.sub.1-C.sub.6)
alkyl, --C(.dbd.O)--O--(C.sub.1-C.sub.6) alkyl and
--C(.dbd.O)--O--(C.sub.1-C.sub.6) substituted alkyl;
[0031] Y is selected from the group consisting of --(CH.sub.2)-- or
a bond:
[0032] wherein said aryl heteroaryl, --O-aryl, --S-aryl,
--O-heteroaryl, and --S-heteroaryl groups may be substituted with
one or more substituents independently selected from the group
consisting of F, Cl, Br, I, --NR5R6, --(C.sub.1-C.sub.6) alkyl,
--(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl,
--O--(C.sub.1-C.sub.6) alkyl, --O--(C.sub.1-C.sub.6) substituted
alkyl, --O--(C.sub.3-C.sub.9) cycloalkyl, --O--(C.sub.3-C.sub.9)
substituted cycloalkyl, -aryl, -aryl-(C.sub.1-C.sub.6) alkyl,
-aryl-O--(C.sub.1-C.sub.6) alkyl, --O-aryl, --O--(C.sub.1-C.sub.4)
alkyl-aryl, heteroaryl, heterocyclyl, --O--(C.sub.1-C.sub.4)
alkyl-heterocycle, and --(S(.dbd.O).sub.2)--(C.sub.1-C.sub.6)
alkyl; and
[0033] m is 1 or 2.
[0034] The present invention also provides a pharmaceutical
composition comprising one or more compounds of formula IIIa, IVa,
IVb, Va, or Vb, and one or more pharmaceutically acceptable
carriers or excipients. The present invention also provides a
pharmaceutical composition comprising a compound of formula IIIa,
IVa, IVb, Va, or Vb, and one or more pharmaceutically acceptable
carriers or excipients.
[0035] The present invention provides a method of treating a cell
proliferative disorder, said method comprising administering to a
subject in need thereof a therapeutically effective amount of a
compound of formula IIIa, IVa, IVb, Va, or Vb, or a
pharmaceutically acceptable salt thereof, or a prodrug, metabolite,
analog or derivative thereof, in combination with a
pharmaceutically acceptable carrier, wherein said cell
proliferative disorder is treated.
[0036] In an embodiment, the cell proliferative disorder is
cancer.
[0037] In a preferred embodiment, the compound is selected from the
group consisting of (+)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione,
(-)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione,
(+)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione, and
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione; or a
pharmaceutically acceptable salt thereof, or a prodrug, metabolite,
analog or derivative thereof.
[0038] The present invention also provides a method of modulating
an activity of c-Met comprising contacting a cell with an effective
amount of a compound of formula IIIa, IVa, IVb, Va, or Vb, or a
pharmaceutically acceptable salt thereof, or a prodrug, metabolite,
analog or derivative thereof, wherein said contacting results in
said modulating an activity of c-Met.
[0039] In an embodiment, the modulating is inhibiting.
[0040] In an embodiment, the compound modulating the activity of
c-Met, without significantly modulating said activity of Protein
Kinase C.
[0041] The present invention also provides a method of selectively
inhibiting an activity of c-Met, without inhibiting an activity of
Protein Kinase C, comprising contacting a cell with an effective
amount of a compound of formula IIIa, IVa, IVb, Va, or Vb, or a
pharmaceutically acceptable salt thereof, or a prodrug, metabolite,
analog or derivative thereof, wherein said contacting results in
said selectively inhibiting said activity of c-Met, without
inhibiting said activity of Protein Kinase C.
[0042] The present invention also provides a method of selectively
inducing cell death in precancerous cells or cancer cells,
comprising contacting a cell with an effective amount of a compound
of formula IIIa, IVa, IVb, Va, or Vb, or a pharmaceutically
acceptable salt thereof, or a prodrug, metabolite, analog or
derivative thereof, wherein said contacting results in said
selectively inducing cell death in said precancerous cells or said
cancer cells.
[0043] The present invention further provides a method of treating
cancer comprising selectively modulating an activity of c-Met, or
both, without significantly modulating an activity of Protein
Kinase C.
[0044] The present invention further provides a method of screening
for a candidate compound for treating cancer, comprising contacting
a cell with a candidate compound, measuring the activity of c-Met,
measuring the activity of Protein Kinase C, and selecting a
candidate compound that is capable of selectively inhibiting the
activity of c-Met, without significantly inhibiting the activity of
Protein Kinase C, wherein said candidate compound that is capable
of selectively inhibiting the activity of c-Met, without
significantly inhibiting the activity of Protein Kinase C, is a
candidate compound for treating cancer. In an embodiment, the
Protein Kinase C activity is measured in vitro.
[0045] Other features and advantages of the present invention are
apparent from the additional descriptions provided herein including
the different examples. The provided examples illustrate different
components and methodology useful in practicing the present
invention. The examples do not limit the claimed invention. Based
on the present disclosure the skilled artisan can identify and
employ other components and methodology useful for practicing the
present invention.
BRIEF DESCRIPTION OF THE FIGURES
[0046] FIG. 1 sets forth the chemical structures of
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij]quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione and
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione.
[0047] FIG. 2 sets forth an effect of
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione on survival
of MDA-MB-231 or Paca-2 cells in vitro.
[0048] FIG. 3 sets forth an effect of
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(+)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione on survival
of MDA-MB-231 cells in vitro.
[0049] FIG. 4 sets forth an effect of
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione on Protein
Kinase C activity in vitro.
[0050] FIG. 5 sets forth inhibition of c-Met phosphorylation and
ERK1/2 phosphorylation by (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
or (+)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione.
[0051] FIG. 6 sets forth an effect of
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione,
administered individually at 160 mg/kg, on the growth of
xenografted MDA-MB-231 human breast cancer tumors in athymic female
nude mice.
[0052] FIG. 7 sets forth an effect of
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione,
administered individually at 80 mg/kg, on the growth of xenografted
MDA-MB-231 human breast cancer tumors in athymic female mice.
[0053] FIG. 8 sets forth an effect of
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione to induce
apoptosis in cancer cells.
[0054] FIG. 9 sets forth an effect of
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione to inhibit
metastatic cancer cell invasion.
[0055] FIG. 10 sets forth an effect of
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione on breast
cancer xenograft model.
[0056] FIG. 11 sets forth an effect of
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione on colon
cancer xenograft model.
[0057] FIG. 12 sets forth an effect of
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione on
pancreatic cancer xenograft model.
DETAILED DESCRIPTION OF THE INVENTION
1. Pyrroloquinolinyl-pyrrole-2,5-diones and
pyrroloquinolinyl-pyrrolidine-2,5-diones
[0058] The present invention provides for
pyrroloquinolinyl-pyrrole-2,5-dione compounds of formula III and
IIIa and their synthesis
##STR00003##
where:
[0059] R1, R2 and R3 are independently selected from the group
consisting of hydrogen, F, Cl, Br, I, --NR5R6, --(C.sub.1-C.sub.6)
alkyl, --(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl,
--O--(C.sub.1-C.sub.6) alkyl, --O--(C.sub.1-C.sub.6) substituted
alkyl, --O--(C.sub.3-C.sub.9) cycloalkyl, and
--O--(C.sub.3-C.sub.9) substituted cycloalkyl, aryl, heteroaryl,
heterocyclyl;
[0060] R4 is independently selected from the group consisting of
hydrogen, --(C.sub.1-C.sub.6) alkyl, --CH.sub.2R7;
[0061] R5, R6 are independently selected from the group consisting
of hydrogen, and --(C.sub.1-C.sub.6) alkyl;
[0062] R7 is independently selected from the group consisting of
--O--P(.dbd.O)(OH).sub.2,
--O--P(.dbd.O)(--OH)(--O--(C.sub.1-C.sub.6) alkyl),
--O--P(.dbd.O)(--O--(C.sub.1-C.sub.6) alkyl).sub.2,
--O--P(.dbd.O)(--OH)(--O--(CH.sub.2)-phenyl),
--O--P(.dbd.O)(--O--(CH.sub.2)-phenyl).sub.2, a carboxylic acid
group, an amino carboxylic acid group and a peptide;
[0063] Q is selected from the group consisting of aryl, heteroaryl,
--O-aryl, --S-aryl, --O-heteroaryl, and --S-heteroaryl;
[0064] X is selected from the group consisting of --(CH.sub.2)--,
--(NR8)-, S, and O;
[0065] R8 is independently selected from the group consisting of
hydrogen, --(C.sub.1-C.sub.6) alkyl, --(C.sub.1-C.sub.6)
substituted alkyl, --(C.sub.3-C.sub.9) cycloalkyl,
--(C.sub.3-C.sub.9) substituted cycloalkyl, and
--O--(C.sub.1-C.sub.9) alkyl, --C(.dbd.O)--O--(C.sub.1-C.sub.6)
alkyl and --C(.dbd.O)--O--(C.sub.1-C.sub.6) substituted alkyl;
[0066] Y is selected from the group consisting of --(CH.sub.2)-- or
a bond;
[0067] wherein said aryl, heteroaryl, --O-aryl, --S-aryl,
--O-heteroaryl, and --S-heteroaryl groups may be substituted with
one or more substituents independently selected from the group
consisting of F, Cl, Br, I, --NR5R6, --(C.sub.1-C.sub.6) alkyl,
--(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl,
--O--(C.sub.1-C.sub.6) alkyl, --O--(C.sub.1-C.sub.6) substituted
alkyl, --O--(C.sub.3-C.sub.9) cycloalkyl, --O--(C.sub.3-C.sub.9)
substituted cycloalkyl, -aryl, -aryl-(C.sub.1-C.sub.6) alkyl,
-aryl-O--(C.sub.1-C.sub.6) alkyl, --O-aryl, --O--(C.sub.1-C.sub.4)
alkyl-aryl, heteroaryl, heterocyclyl, --O--(C.sub.1-C.sub.4)
alkyl-heterocycle, and --(S(.dbd.O).sub.2)--(C.sub.1-C.sub.6)
alkyl; and
[0068] m is 1 or 2.
[0069] For the compound of formula IIIa, Q is selected from the
group consisting of aryl, heteroaryl, --O-aryl, --S-aryl,
--O-heteroaryl, and --S-heteroaryl, provided that when R4 is
hydrogen, (C.sub.3-C.sub.4) cycloalkyl, or (C.sub.1-C.sub.4) alkyl,
Q is not 3-indolyl or substituted 3-indolyl.
[0070] The present invention also provides for
pyrroloquinolinyl-pyrrolidine-2,5-dione compounds of formula IVa,
IVb, Va, or Vb, and methods of preparing the compounds of formulas
IVa, IVb, Va, and Vb,
##STR00004##
where:
[0071] R1, R2 and R3 are independently selected from the group
consisting of hydrogen, F, Cl, Br, I, --NR5R6, --(C.sub.1-C.sub.6)
alkyl, --(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl,
--O--(C.sub.1-C.sub.6) alkyl, --O--(C.sub.1-C.sub.6) substituted
alkyl, --O--(C.sub.3-C.sub.9) cycloalkyl, and
--O--(C.sub.3-C.sub.9) substituted cycloalkyl, aryl, heteroaryl,
heterocyclyl;
[0072] R4 is independently selected from the group consisting of
hydrogen, --(C.sub.1-C.sub.6) alkyl. --CH.sub.2R7;
[0073] R5, R6 are independently selected from the group consisting
of hydrogen, and --(C.sub.1-C.sub.6) alkyl;
[0074] R7 is independently selected from the group consisting of
--O--P(.dbd.O)(OH).sub.2,
--O--P(.dbd.O)(--OH)(--O--(C.sub.1-C.sub.6) alkyl),
--O--P(.dbd.O)(--O--(C.sub.1-C.sub.6) alkyl).sub.2,
--O--P(.dbd.O)(--OH)(--O--(CH.sub.2)-phenyl).
--O--P(.dbd.O)(--O--(CH.sub.2)-phenyl).sub.2, a carboxylic acid
group, an amino carboxylic acid group and a peptide;
[0075] Q is selected from the group consisting of aryl, heteroaryl,
--O-aryl, --S-aryl, --O-heteroaryl, and --S-heteroaryl;
[0076] X is selected from the group consisting of --(CH.sub.2)--,
--(NR8)-, S, and O;
[0077] R8 is independently selected from the group consisting of
hydrogen, --(C.sub.1-C.sub.6) alkyl, --(C.sub.1-C.sub.6)
substituted alkyl, --(C.sub.3-C.sub.9) cycloalkyl,
--(C.sub.3-C.sub.9) substituted cycloalkyl, and
--O--(C.sub.1-C.sub.6) alkyl, --C(.dbd.O)--O--(C.sub.1-C.sub.6)
alkyl and --C(.dbd.O)--O--(C.sub.1-C.sub.6) substituted alkyl;
[0078] Y is selected from the group consisting of --(CH.sub.2)-- or
a bond;
[0079] wherein said aryl, heteroaryl, --O-aryl, --S-aryl,
--O-heteroaryl, and --S-heteroaryl groups may be substituted with
one or more substituents independently selected from the group
consisting of F, Cl, Br, I, --NR5R6, --(C.sub.1-C.sub.6) alkyl,
--(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl,
--O--(C.sub.1-C.sub.6) alkyl, --O--(C.sub.1-C.sub.6) substituted
alkyl, --O--(C.sub.3-C.sub.9) cycloalkyl, --O--(C.sub.3-C.sub.9)
substituted cycloalkyl, -aryl, -aryl-(C.sub.1-C.sub.6) alkyl,
-aryl-O--(C.sub.1-C.sub.6) alkyl, --O-aryl, --O--(C.sub.1-C.sub.4)
alkyl-aryl, heteroaryl, heterocyclyl, --O--(C.sub.1-C.sub.4)
alkyl-heterocycle, and --(S(.dbd.O).sub.2)--(C.sub.1-C.sub.6)
alkyl; and
[0080] m is 1 or 2.
1.1. Definitions
[0081] The term "alkyl" refers to radicals containing carbon and
hydrogen, without unsaturation. Alkyl radicals can be straight or
branched. Exemplary alkyl radicals include, without limitation,
methyl, ethyl, propyl, isopropyl, hexyl, t-butyl, sec-butyl and the
like. Akyl groups may be denoted by a range, thus, for example, a
(C.sub.1-C.sub.6) alkyl group is an alkyl group having from one to
six carbon atoms in the straight or branched alkyl backbone.
Substituted and unsubstituted alkyl groups may independently be
(C.sub.1-C.sub.5) alkyl, (C.sub.1-C.sub.6) alkyl,
(C.sub.1-C.sub.10) alkyl, (C.sub.3-C.sub.10) alkyl, or
(C.sub.5-C.sub.10) alkyl. Unless expressly stated, the term "alkyl"
does not include "cycloalkyl."
[0082] A "cycloalkyl" group refers to a cyclic alkyl group having
the indicated number of carbon atoms in the "ring portion," where
the "ring portion" may consist of one or more ring structures
either as fused, spiro, or bridged ring structures. For example, a
C3 to C6 cycloalkyl group (e.g., (C.sub.3-C.sub.6) cycloalkyl) is a
ring structure having between 3 and 6 carbon atoms in the ring.
When no range is given, then cycloalkyl has between three and nine
carbon atoms ((C.sub.3-C.sub.9) cycloalkyl) in the ring portion.
Exemplary cycloalkyl groups include, but are not limited to
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and
adamantyl. Preferred cycloalkyl groups have three, four, five, six,
seven, eight, nine, or from three to nine carbon atoms in the ring
structure.
[0083] The term substituted alkyl and substituted cycloalkyl, refer
to alkyl and cycloalkyl groups, as defined above, substituted with
one or more substituents independently selected from the group
consisting of fluorine, aryl, heteroaryl, --O--(C.sub.1-C.sub.6)
alkyl, and --NR5R6, where R5 and R6 are independently selected from
the group consisting of hydrogen and --(C.sub.1-C.sub.6) alkyl.
[0084] The term "aryl" refers to an aromatic carbocyclic group,
having one, two, or three aromatic rings. Exemplary aryl groups
include, without limitation, phenyl, naphthyl, and the like. Aryl
groups include one, two, or three aromatic rings structures fused
with one or more additional nonaromatic carbocyclic or hetercyclic
rings having from 4-9 members. Examples of fused aryl groups
include benzocyclobutanyl, indanyl, tetrahydronapthylenyl,
1,2,3,4-tetrahydrophenanthrenyl, tetrahydroanthracenyl,
1,4-dihydro-1,4-methanonaphthalenyl, benzodioxolyl.
[0085] The term "heteroaryl" refers to a heteroaromatic
(heteroaryl) group having one, two, or three aromatic rings
containing from 1-4 heteroatoms (such as nitrogen, sulfur, or
oxygen) in the aromatic ring. Heteroaryl groups include one, two,
or three aromatic rings structures containing from 1-4 heteroatoms
fused with one or more additional nonaromatic rings having from 4-9
members. Heteroaryl groups containing a single type of heteroatom
in the aromatic ring are denoted by the type of hetero atom they
contain, thus, nitrogen-containing heteroaryl, oxygen-containing
heteroaryl and sulfur-containing heteroaryl denote heteroaromatic
groups containing one or more nitrogen, oxygen or sulfur atoms
respectively. Exemplary heteroaryl groups include, without
limitation, pyridyl, pyrimidinyl, triazolyl, quinolyl,
quinazolinyl, thiazolyl, benzo[b]thiophenyl, furanyl, imidazolyl,
indolyl, and the like.
[0086] The terms "heterocyclyl" or "heterocycle" refers to either
saturated or unsaturated, stable nonaromatic ring structures that
may be fused, spiro or bridged to form additional rings. Each
heterocycle consists of one or more carbon atoms and from one to
four heteroatoms selected from the group consisting of nitrogen,
oxygen and sulfur. "Heterocyclyl" or "heterocycle" include stable
nonaromatic 3-7 membered monocyclic heterocyclic ring structures
and 8-11 membered bicyclic heterocyclic ring structures. A
heterocyclyl radical may be attached at any endocyclic carbon or
nitrogen atom that results in the creation of a stable structure.
Preferred heterocycles include 3-7 membered monocyclic heterocycles
(more preferably 5-7-membered monocyclic heterocycles) and 8-10
membered bicyclic heterocycles. Examples of such groups include
piperidinyl, piperazinyl, pyranyl, pyrrolidinyl, morpholinyl,
thiomorpholinyl, oxopiperidinyl, oxopyrrolidinyl, oxoazepinyl,
azepinyl, isoxozolyl, tetrahydropyranyl, tetrahydrofuranyl,
dioxolyl, dioxinyl, oxathiolyl, dithiolyl, sulfolanyl, dioxanyl,
dioxolanyl, tetahydrofurodihydrofuranyl,
tetrahydropyranodihydro-furanyl, dihydropyranyl,
tetrahydrofurofuranyl, tetrahydropyranofuran, quinuclidinyl
(1-azabicyclo[2.2.2]octanyl) and tropanyl
(8-methyl-8-azabicyclo[3.2.1]octanyl).
[0087] For the purpose of the Q substituent, the term "substituted
3-indolyl" refers to a 3-indolyl group substituted with one or more
substituents selected from the group consisting of: F, Cl. Br, I,
--NR5R6, --(C.sub.1-C.sub.6) alkyl, --(C.sub.1-C.sub.6) substituted
alkyl, --(C.sub.3-C.sub.9) cycloalkyl, --(C.sub.3-C.sub.9)
substituted cycloalkyl, --O--(C.sub.1-C.sub.9) alkyl,
--O--(C.sub.1-C.sub.6) substituted alkyl, --O--(C.sub.1-C.sub.9)
cycloalkyl, --O--(C.sub.3-C.sub.9) substituted cycloalkyl, -aryl,
-aryl-(C.sub.1-C.sub.6) alkyl, -aryl-O--(C.sub.1-C.sub.6) alkyl,
--O-aryl, --O--(C.sub.1-C.sub.4) alkyl-aryl, heteroaryl,
heterocyclyl, --O--(C.sub.1-C.sub.4) alkyl-heterocycle, and
--(S(.dbd.O).sub.2)--(C.sub.1-C.sub.6) alkyl; where R5, R6 are
independently selected from the group consisting of hydrogen, and
--(C.sub.1-C.sub.6) alkyl.
[0088] For the purposes of the R7 substituent, the term "carboxylic
acid group" refers to a group of the form
--O--C(.dbd.O)--(C.sub.1-C.sub.6) alkyl,
--O--C(.dbd.O)--(C.sub.3-C.sub.9) cycloalkyl, --O--C(.dbd.O)-aryl,
--O--C(.dbd.O)-heteroaryl, --O--C(.dbd.O)-heterocycle,
--O--C(.dbd.O)--(C.sub.1-C.sub.6) alkyl-aryl,
--O--C(.dbd.O)--(C.sub.1-C.sub.6) alkyl-heteroaryl, or
--O--C(.dbd.O)--(C.sub.1-C.sub.6) alkyl-heterocycle. Included in
"carboxylic acid group" are groups of the form
--O--C(.dbd.O)--(C.sub.1-C.sub.6) alkyl,
--O--C(.dbd.O)--(C.sub.3-C.sub.9) cycloalkyl, --O--C(.dbd.O)-aryl,
--O--C(.dbd.O)-heteroaryl, --O--C(.dbd.O)-heterocycle,
--O--C(.dbd.O)--(C.sub.1-C.sub.6) alkyl-aryl,
--O--C(.dbd.O)--(C.sub.1-C.sub.6) alkyl-heteroaryl, or
--O--C(.dbd.O)--(C.sub.1-C.sub.6) alkyl-heterocycle substituted
with one or more substituent independently selected from the group
consisting of: F, Cl, Br, I, --OH, --SH, --NR5R6,
--(C.sub.1-C.sub.6) alkyl, --(C.sub.1-C.sub.6) substituted alkyl,
--(C.sub.3-C.sub.9) cycloalkyl, --(C.sub.3-C.sub.9) substituted
cycloalkyl, --O--(C.sub.1-C.sub.6) alkyl, --O--(C.sub.1-C.sub.6)
substituted alkyl, --S--(C.sub.1-C.sub.6) alkyl,
--O--(C.sub.3-C.sub.9) cycloalkyl, --O--(C.sub.3-C.sub.9)
substituted cycloalkyl, -aryl, --O-aryl, --O--(C.sub.1-C.sub.4)
alkyl-aryl, heteroaryl, heterocyclyl, --O--(C.sub.1-C.sub.4)
alkyl-heterocycle, --(S(.dbd.O).sub.2)--(C.sub.1-C.sub.6) alkyl,
--NH--C(.dbd.NH)--NH.sub.2 (i.e., guanido), --COOH, and
--C(.dbd.O)--NR5R6, where R5 and R6 are independently selected from
the group consisting of hydrogen, and --(C.sub.1-C.sub.6) alkyl. In
addition, for the purposes of the R7 substituent the term "amino
carboxylic acid group" refers to a carboxylic acid group, including
carboxylic acid groups substituted with one or more of the
above-stated substituents, which bears one or more independently
selected amino groups of the form --NR5R6 where R5 and R6 are
independently selected from the group consisting of hydrogen and
(C1-C6) alkyl.
[0089] In one embodiment of this invention, R7 is an alpha amino or
imino acid, including but not limited to alanine, arginine,
asparagine, aspartic acid, cysteine, glutamine, glutamic acid,
glycine, histidine, isoleucine, leucine, lysine, methionine,
phenylalanine, proline, serine, threonine, tryptophan, tyrosine,
valine or stereoisomers or racemic mixtures thereof. In another
embodiment the of the invention R7 is alpha amino or imino acid
selected from the group consisting of L-alanine, L-arginine,
L-asparagine, L-aspartic acid, L-cysteine, L-glutamine, L-glutamic
acid, L-glycine, L-histidine, L-isoleucine, L-leucine, L-lysine,
L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine,
L-tryptophan, L-tyrosine, L-valine.
[0090] For the purposes of the R7 substituent, the term "peptide"
refers to a dipeptide, tripeptide, tetrapeptide or pentapeptide,
which release two, three, four, or five amino or imino acids (e.g.,
proline) respectively upon hydrolysis. For the purpose of R7,
peptides are linked to the remainder of the molecule through an
ester linkage. In one embodiment, peptides of R7 are comprised of
alpha amino or imino acid, including but not limited to alanine,
arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic
acid, glycine, histidine, isoleucine, leucine, lysine, methionine,
phenylalanine, proline, serine, threonine, tryptophan, tyrosine,
valine or stereoisomers or racemic mixtures thereof; and in a more
preferred version of this embodiment, the carboxyl group involved
in the ester linkage is the carboxyl terminal COOH group of the
peptide, as opposed to a side chain carboxyl. In another embodiment
the of the invention R7 is alpha amino or imino acid selected from
the group consisting of L-alanine, L-arginine, L-asparagine,
L-aspartic acid, L-cysteine, L-glutamine, L-glutamic acid,
L-glycine, L-histidine, L-isoleucine, L-leucine, L-lysine,
L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine,
L-tryptophan, L-tyrosine, and L-valine; and in a more preferred
version of this preferred embodiment, the carboxyl group involved
in the ester linkage is the carboxyl terminal COOH group of the
peptide, as opposed to a side chain carboxyl.
1.2. Preferred Compounds
[0091] Included in the preferred embodiments are compounds of
formula III, IVa, IVb, Va, or Vb, wherein Q is selected from the
group consisting of aryl, heteroaryl, --O-aryl, --S-aryl,
--O-heteroaryl, and --S-heteroaryl, provided that Q is not
3-indolyl or a substituted 3-indolyl. In other preferred
embodiments Q is selected from the group consisting of aryl,
heteroaryl, --O-aryl, --S-aryl, --O-heteroaryl, and --S-heteroaryl,
provided that when R4 is hydrogen, cycloalkyl, or alkyl, Q is not
3-indolyl or a substituted 3-indolyl. In still other preferred
embodiments Q is selected from the group consisting of aryl,
heteroaryl, --O-aryl, --S-aryl, --O-heteroaryl, and --S-heteroaryl,
provided that when R4 is hydrogen, (C.sub.3-C.sub.4) cycloalkyl, or
(C.sub.1-C.sub.4) alkyl, Q is not 3-indolyl or substituted
3-indolyl. In another preferred embodiment Q is 3-indolyl or a
substituted 3-indolyl provided that R4 is not hydrogen, cycloalkyl,
or alkyl. In still another preferred embodiment Q is 3-indolyl or a
substituted 3-indolyl provided that R4 is not hydrogen,
(C.sub.3-C.sub.4) cycloalkyl, or (C.sub.1-C.sub.4) alkyl.
[0092] Other preferred embodiments include compounds of formulas
IIIa, IVa, IVb, Va, or Vb where R4 is --CH.sub.2R7. These compounds
may serve as prodrug forms of the corresponding compounds of
formulas IIIa, IVa, IVb, Va, or Vb where R4 is H. The prodrug form
is cleaved by hydrolysis to release the corresponding compound
where R4 is H. The hydrolysis may occur by enzymatic or
nonenzymatic routes that produce the corresponding hydroxymethylene
derivative, which upon subsequent hydrolysis, result in the release
of compounds where R4 is H. In one such preferred embodiment R4 is
--CH.sub.2R7, where R7 is --O--P(.dbd.O)(OH),
--O--P(.dbd.O)(--OH)(--O--(C.sub.1-C.sub.6)alkyl), or
--O--P(.dbd.O)(--O--(C.sub.1-C.sub.6)alkyl).sub.2. In one
embodiment where R7 is
--O--P(.dbd.O)(--O--(C.sub.1-C.sub.6)alkyl).sub.2, the alkyl groups
are independently selected. In another preferred embodiment, R4 is
--CH.sub.2R7, where R7 is a carboxylic acid group or an amino
carboxylic acid group. In still another preferred embodiment R7 is
a peptide; where in a more preferred embodiment the peptide is
linked through an ester bond formed with the carboxyl terminal COOH
group of the peptide chain to the remainder of the compound. In
other preferred separate and independent embodiments of compounds
of formulas IIIa, IVa, IVb, Va, or Vb where R4 is --CH.sub.2R7 and
R7 is a peptide, the peptide may be a dipeptide, a tripeptide, a
tetrapeptide or a pentapeptide. Preferred amino acid compositions
for peptides of the R7 functionality are described above.
[0093] Embodiments of compounds of formulas III, IIIa, IVa, IVb,
Va, or Vb include those where X is selected from the group
consisting of --(NR8)-, S, and O, where R8 is independently
selected from the group consisting of hydrogen, --(C.sub.1-C.sub.6)
alkyl, --(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl, and
--O--(C.sub.1-C.sub.6) alkyl. Other embodiments of compounds of
formulas III, IIIa, IVa, IVb, Va, or Vb include those where X is
--CH.sub.2--. In other embodiments of compounds of formulas III,
IIIa, IVa, IVb, Va, or Vb, X is oxygen (O). In other embodiments of
compounds of formulas III, IIIa, IVa, IVb, Va, or Vb, X is sulfur
(S). In still other embodiments of compounds of formulas III, IIIa,
IVa, IVb, Va, or Vb, X is --(NR8)-, where R8 is independently
selected from the group consisting of hydrogen, --(C.sub.1-C.sub.6)
alkyl, --(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl, and
--O--(C.sub.1-C.sub.6) alkyl.
[0094] Other preferred embodiments of the invention include
compounds of formula IIIa, where Q is a heteroaryl or an optionally
substituted heteroaryl group. In four separate alternative
preferred embodiments of compounds of formula IIIa, Q is an
optionally substituted monocyclic heteroaryl group, an optionally
substituted bicyclic heteroaryl group, an optionally substituted
bicyclic heteroaryl group with the proviso that the bicyclic
heteroaryl group is not an indolyl group or a substituted indolyl,
or an optionally substituted tricyclic heteroaryl group. Optional
substituents, when present, are independently selected from those
recited for aryl, heteroaryl, --O-aryl, --S-aryl, --O-heteroaryl,
and --S-heteroaryl.
[0095] Included in the preferred embodiments of the invention are
compounds of formulas IVa, IVb, Va, or Vb, where Q is a heteroaryl
or an optionally substituted heteroaryl group. In four separate
alternative preferred embodiments of compounds of formulas IVa,
IVb, Va, or Vb, Q is an optionally substituted monocyclic
heteroaryl group, an optionally substituted bicyclic heteroaryl
group, an optionally substituted bicyclic heteroaryl group with the
proviso that the bicyclic heteroaryl group is not indolyl, or an
optionally substituted tricyclic heteroaryl group. In a more
preferred embodiment, Q is an optionally substituted
nitrogen-containing heteroaryl group. In a related embodiment, Q is
an optionally substituted indolyl. Optional substituents, when
present are independently selected from those recited for aryl,
heteroaryl, --O-aryl, --S-aryl, --O-heteroaryl, and
--S-heteroaryl.
[0096] Preferred embodiments of the invention include mixtures of
compounds of formulas IVa and IVb, including racemic mixtures. In
another preferred embodiment, the compounds of formula IVa and IVb
are the separate enantiomers of
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione. In this
embodiment the preparation of (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione is
prepared as a mixture beginning with the starting materials
1,2,3,4-tetrahydroquinoline and indole-3-acetamide. The
1,2,3,4-tetrahydroquinoline is converted into
5,6-dihydro-4H-pyrrolo [3,2,1-ij] quinolin-1-yl) oxoacetic acid
methyl ester as described in Example 1, steps 1-5. The 5,
6-dihydro-4H-pyrrolo [3,2,1-ij] quinolin-1-yl) oxoacetic acid
methyl ester is reacted with indole-3-acetamide as described in
Example 1, step 6, to yield 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione. The mixture of
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione is then
prepared by catalytic hydrogenation as described in Example 2 using
Procedure B.
[0097] Preferred embodiments of the invention also include mixtures
of compounds of formulas Va and Vb, including racemic mixtures. In
another preferred embodiment, the compounds of Va and Vb are the
separate enantiomers of (.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione.
In this embodiment, the compounds are prepared as a mixture by
first preparing (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione, as described
above. The mixture of cis compounds is then treated with a mixture
of potassium tert-butoxide in tert-butanol to obtain a mixture of
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione as described
in Example 3.
[0098] All stereoisomers of the compounds of the instant invention
are contemplated, either in admixture or in pure or substantially
pure form, including crystalline forms of racemic mixtures and
crystalline forms of individual isomers. The definition of the
compounds according to the invention embraces all possible
stereoisomers (e.g., the R and S configurations for each asymmetric
center) and their mixtures. It very particularly embraces the
racemic forms and the isolated optical isomers having a specified
activity. The racemic forms can be resolved by physical methods,
such as, for example, fractional crystallization, separation or
crystallization of diastereomeric derivatives, separation by chiral
column chromatography or supercritical fluid chromatography. The
individual optical isomers can be obtained from the racemates by
conventional methods, such as, for example, salt formation with an
optically active acid followed by crystallization. Furthermore, all
geometric isomers, such as E- and Z-configurations at a double
bond, are within the scope of the invention unless otherwise
stated. Certain compounds of this invention may exist in tautomeric
forms. All such tautomeric forms of the compounds are considered to
be within the scope of this invention unless otherwise stated. The
present invention also includes one or more regioisomeric mixtures
of an analog or derivative.
[0099] As used herein, the term "salt" is a pharmaceutically
acceptable salt and can include acid addition salts including
hydrochlorides, hydrobromides, phosphates, sulphates, hydrogen
sulphates, alkylsulphonates, arylsulphonates, acetates, benzoates,
citrates, maleates, fumarates, succinates, lactates, and tartrates;
alkali metal cations such as Na.sup.+, K.sup.+, Li.sup.+, alkali
earth metal salts such as Mg or Ca, or organic amine salts.
[0100] As used herein, the term "metabolite" means a product of
metabolism of a compound of the present invention, or a
pharmaceutically acceptable salt, analog or derivative thereof,
that exhibits a similar activity in vivo to said compound of the
present invention.
[0101] As used herein, the term "prodrug" means a compound of the
present invention covalently linked to one or more pro-moieties,
such as an amino acid moiety or other water solubilizing moiety. A
compound of the present invention may be released from the
pro-moiety via hydrolytic, oxidative, and/or enzymatic release
mechanisms. In an embodiment, a prodrug composition of the present
invention exhibits the added benefit of increased aqueous
solubility, improved stability, and improved pharmacokinetic
profiles. The pro-moiety may be selected to obtain desired prodrug
characteristics. For example, the pro-moiety, e.g., an amino acid
moiety or other water solubilizing moiety such as phosphate within
R4, may be selected based on solubility, stability,
bioavailability, and/or in vivo delivery or uptake.
2. The Synthesis of Pyrroloquinolinyl-pyrrole-2,5-diones and
pyrroloquinolinyl-pyrrolidine-2,5-diones
[0102] Standard synthetic methods and procedures for the
preparation of organic molecules and functional group
transformations and manipulations including the use of protective
groups can be obtained from the relevant scientific literature or
from standard reference textbooks in the field. Although not
limited to any one or several sources, recognized reference
textbooks of organic synthesis include: Smith, M. B.; March, J.
March's Advanced Organic Chemistry: Reactions, Mechanisms, and
Structure, 5.sup.th ed.; John Wiley & Sons: New York, 2001; and
Greene, T. W.; Wuts, P. G. M. Protective Groups in Organic
Synthesis, 3.sup.rd; John Wiley & Sons: New York, 1999. The
following descriptions of synthetic methods are designed to
illustrate, but not limit, general procedures for the preparation
of compounds of the invention.
2.1 General Procedures for the Synthesis of
pyrroloquinolinyl-pyrrole-2,5-dione and
pyrroloquinolinyl-pyrrolidine-2,5-diones where R4 is hydrogen
[0103] The present invention provides for
pyrroloquinolinyl-pyrrole-2,5-dione compounds of formula IIIa, IVa,
IVb, Va, or Vb. The preparation of compounds of formulas III, IIIa,
IVa, IVb, Va, and Vb may be achieved by a series of reactions
commencing with the reaction of a 5, 6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl) oxoacetic acid ester of formula I with an
amide of formula II, to form a 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione of formula III,
including compounds of formula IIIa, where R4 is hydrogen, as shown
in Scheme 1.
##STR00005##
2.1.1. Synthesis of 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1-indol-3-yl) pyrrole-2, 5-diones of formula III
where R4 is hydrogen
[0104] The condensation of an ester of formula I and a compound of
formula II to produce compounds of formula III, including compounds
of formula IIIa, where R4 is hydrogen is conducted in any suitable
anhydrous polar aprotic solvent including, but not limited to,
tetrahydrofuran (THF), tetrahydropyran, diethyl ether and the like
in the presence of base. For the purposes of the reaction, suitable
esters of formula I include, but are not limited to, alkyl esters
where R9 is a (C1-C4) alkyl group, and preferred esters include the
methyl and ethyl esters. Suitable bases for the reaction include
alkaline metal salts of low molecular weight alkyl alcohols,
including, but not limited to, alkaline metal salts of methanol,
ethanol, propanol, isopropanol, n-butanol, isobutanol, and
tert-butanol. Preferred alkaline metal salts of low molecular
weight alkyl alcohols include sodium and potassium salts, with
potassium tert-butoxide (tBuOK) being the preferred base. Typically
the reactions are conducted at 0.degree. C. for 2 hours, however,
both the time and temperature may be altered depending upon the
specific substituents present on compounds of formula I and II, and
the solvent employed. The reaction temperature may be varied from
-78.degree. C. to 37.degree. C. and is preferably from -35.degree.
C. to 25.degree. C., or more preferably from -15.degree. C. to
10.degree. C. Reaction times will generally vary inversely with the
temperature employed, suitable times from about 15 minutes to 24
hours may be employed, more preferably, 30 minutes to 12 hours, and
more preferably 1 to 6 hours.
2.1.2. Preparation of Compounds of Formulas IVa, IVb, Va and Vb
where R4 is hydrogen
[0105] Reduction of compounds of formulas III and IIIa, where R4 is
hydrogen to yield the corresponding 3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-diones
having formula IVa, IVb, Va, or Vb may be conducted employing a
variety of procedures including, but not limited to, reduction with
zinc-mercury (Procedure A), catalytic hydrogenation (Procedure B),
and reduction with magnesium in methanol (Procedure C). As
indicated in Scheme 1, depending on the reduction reaction and
conditions chosen, the reaction will yield principally compounds of
formulas IVa and IVb, or principally compounds of formulas Va and
Vb, or alternatively a mixture of compounds of formulas IVa, IVb,
Va, and Vb.
[0106] Mixtures of compounds of formulas IVa, IVb, Va, and Vb may
be prepared by the direct reduction of compounds of formulas III or
IIIa with a zinc-mercury reducing agent. The reaction is generally
carried out with fresh reducing agent prepared by mixing Zn powder
with HgCl.sub.2 deionized water followed by acidification with HCl.
After drying, the solid reducing agent (zinc-mercury) is suitable
for reduction of compounds of formulas III or IIIa in refluxing dry
ethanol under a dry HCl gas atmosphere as described in Example 2,
Procedure A, for the reduction of 3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione.
[0107] An alternative method of preparing pyrrolidine-2, 5-diones
is catalytic hydrogenation, which yields a mixture consisting
principally of the (.+-.)-cis pyrrolidine-2, 5-diones of formulas
IVa and IVb. Catalytic hydrogenation of compounds of formulas III
or IIIa may be conducted in an anhydrous alcohol over a noble metal
catalyst under 1 atmosphere of hydrogen for 48 hours. A variety of
low molecular weight alkyl alcohols may be employed to conduct the
reduction, including n-propyl alcohol, isopropyl alcohol, ethanol
or methanol. Preferably the alcohol is ethanol or methanol, and
most preferably methanol. A noble metal catalyst (e.g., platinum,
palladium, rhodium, ruthenium etc.) on charcoal is preferred for
the reduction of compounds of formulas Ill or IIIa. In more
preferred embodiments, the noble metal catalyst is palladium on
activated charcoal. While reduction compounds of formulas IIIa or
III under 1 atmosphere of hydrogen at room temperature (25.degree.
C.) for 12-48 hours is generally suitable for preparation of
pyrrolidine-2, 5-diones, the pressure of hydrogen, reaction time,
and the reaction temperature may be varied. Catalytic hydrogenation
of 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione is described in
Example 2, Procedure B.
[0108] Pyrrole-2,5-diones of formula IIIa or III may be reduced to
yield a mixture of compounds of formulas Va and Vb by the reduction
in anhydrous alcohol with a metal reducing agent. Preferred metals
include sodium, calcium and magnesium, with magnesium as a more
preferred metal reducing agent. The reaction is typically carried
out under an inert atmosphere of nitrogen for 30 minutes to 2 hours
by refluxing a compound of formula III or formula IIIa in an
alcohol selected from the group consisting of methanol, ethanol,
n-propanol, and isopropanol with magnesium turnings. In preferred
embodiments the reaction is conducted for about 40 minutes in
methanol as described in Example 2, Procedure C, for the
preparation of (.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione.
[0109] Compounds of IVa and/or IVb, which have the pyrrolidine ring
substituents in the cis configuration, may be converted into a
mixture of compounds of Va and Vb, where the substituents are in
the trans configuration, or into a mixture of all four isomers of
formulas IVa, IVb, Va, and Vb by treatment with base in a polar
protic solvent. Typically the reaction employs an alkaline metal
salt of a (C1-C4) alkyl alcohol in an alcohol solvent (e.g., sodium
or potassium methoxide in methanol, sodium or potassium ethoxide in
ethanol, sodium or potassium tert-butoxide in tert-butanol), with
potassium tert-butoxide in tert-butanol as the preferred alkaline
metal salt and solvent mixture. Reactions are generally conducted
from 0.degree. C. to the reflux temperature of the reaction mixture
for 4 to 48 hours. In more preferred embodiments, the reaction are
conducted from room temperature (25.degree. C.) to the reflux
temperature of the mixture for 8 to 24 hours, and in an even more
preferred embodiment, the reaction is conducted at about 50.degree.
C. in a mixture of potassium tert-butoxide in tert-butanol for
about 16 hours. Short reaction times and low temperatures favor
formation of mixtures still containing compounds IVa and/or
IVb.
2.1.3. Introduction of Aryl or Heteroaryl Substituents into
Compounds of III, IIIa, IVa, IVb, Va, and Vb
[0110] The introduction of additional substituted and unsubstituted
aryl or heteroaryl substituents on to aromatic rings of compounds
of formulas III, IIIa, IVa, IVb, Va, or Vb may be accomplished by
the reaction of a substituted or unsubstituted aryl or heteroaryl
boronic acid with an aromatic halogen substituent on a compound of
formula III, IIIa, IVa, IVb, Va, or Vb. The reaction is typically
carried out by heating a mixture of a compound of formula III,
IIIa, IVa, IVb, Va, or Vb bearing an aryl or heteroaryl bromide or
iodide, more preferably an arylbromide or hetroarylbromide, with an
aryl or heteroaryl boronic acid in the presence of
tetrakistriphenylphosphine palladium in a solvent mixture
consisting of 5 parts toluene, 5 parts ethanol, 1 part saturated
NaHCO.sub.3, and 2 parts water to 100.degree. C. under nitrogen for
5 hours. After cooling to room temperature, the mixture is
extracted with ethyl acetate and concentrated. The residue is
purified by silica gel chromatography. In a preferred embodiment,
the halogenated compound of formula III, IIIa, IVa, IVb, Va, or Vb
bears the halogen on an aryl or heteroaryl group Q functionality
resulting in the introduction of substituted aryl or heteroaryl
group donated by the boronic acid on to the Q substituent. In a
more preferred embodiment, the Q functionality is a brominated
aromatic or heteroaromatic Q functionality. In another more
preferred embodiment the halogenated Q functionality reacted with
the boronic acid is a halogenated 3-indolyl. Examples 31-34
describe the introduction of substituted and unsubstituted aromatic
groups into compounds of formula Va and Vb employing a brominated Q
functionality where Q is a brominated 3-indolyl.
[0111] Aromatic and heteroaromatic boronic acids including
2-thienylboronic acid, 3-thienylboronic acid, and 2-naphthylboronic
acid are available from a variety of commercial sources including
Sigma-Aldrich (St. Louis, Mo.). Alternatively aromatic and
heteroaromatic boronic acids may be prepared from the corresponding
aryl or heteroaryl bromides by reaction with triisopropyl borate in
the presence of n-butyllithium followed by quenching with aqueous
HCl. (See, e.g., W. Li, et. al., J. Organic Chem. 67: 5394-97
(2002) and C. M. Marson, et. al., Tetrahedron 59: 4377-81
(2003).
2.1.4. Preparation of Compounds of Formulas III, IIIa, IVa IVb, Va,
and Vb where R4 is --CH.sub.2R7
[0112] Compounds of formula III, IIIa, IVa, IVb, Va, or Vb, where
R4 is hydrogen, can be converted into compounds of formula III,
IIIa, IVa, IVb, Va, or Vb where R4 is --CH.sub.2R7. The conversion
begins with the preparation of the hydroxymethylene derivative of
the compounds as indicated in the partial structures shown in
Scheme 2.
##STR00006##
[0113] Preparation of the hydroxymethylene derivatives is
accomplished by reaction of a compound of formula III, IIIa, IVa,
IVb, Va, or Vb where R4 is H with aqueous formaldehyde in
tetrahydrofuran (THF). Typical reaction conditions employ equal
volumes of THF and 37% formaldehyde in water with the reaction
stirred for 14-16 hours at room temperature. Reaction times and
temperatures may vary from 1 hour to 48 hours and the temperature
may be from 0.degree. C. to 50.degree. C. or more preferably from
10.degree. C. to 37.degree. C. Upon completion the reaction is
partitioned between water and an organic solvent, typically ethyl
acetate. The organic layer is dried over sodium sulfate,
concentrated, and subject to chromatography on silica gel as
necessary to yield the hydroxymethylene product. The preparation of
the hydroxymethylene derivative of 3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione,
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-1-hydroxymethyl-4-(1H-i-
ndol-3-yl)-pyrrolidine-2,5-dione, is described in Example 56, step
1.
[0114] Compounds of formulas III, IIIa, IVa, IVb, Va, or Vb where
R4 is --CH.sub.2R7 and R7 is phosphate (--O--P(.dbd.O)(OH).sub.2),
monoalkyl phosphate (e.g.,
--O--P(.dbd.O)(--OH)(--O--(C.sub.1-C.sub.6) alkyl)), dialkyl
phosphate (e.g., --O--P(.dbd.O)(--O--(C.sub.1-C.sub.6)
alkyl).sub.2) a monobenzylphosphate ester (--O--P(.dbd.O)(--OH)
(--O--(CH.sub.2)-phenyl)), or a dibenzylphosphate ester
(--O--P(.dbd.O)(--O--(CH.sub.2)-phenyl).sub.2) may be prepared from
the desired hydroxymethylene derivative and a suitably substituted
phosphoric acid by any reaction suitable for the formation of a
phosphate ester bond between the phosphoric acid compound and the
hydroxymethylene derivative. In a preferred method, the formation
of phosphate esters is conducted by reaction of a hydroxymethylene
derivative of a compound of formula III, IIIa, IVa, IVb, Va, or Vb
with a suitably protected phosphoramidate followed by deprotection.
Reactions with the desired phosphoramidate are typically conducted
at room temperature in anhydrous THE. Following the addition of the
phosphoramidate, the reaction is treated with tetrazole (3% in
acetonitrile) and stirred 5 minutes to 1 hour, after which the
reaction is cooled to -78.degree. C. The cooled reaction is treated
with m-chloroperbenzoic acid, and after stirring at -78.degree. C.
for 5 minutes, the reaction is warmed to room temperature and
stirred for 5 minutes further. Following the removal of solvent,
the product is purified by flash chromatography on silica gel using
ethyl acetate hexane. The protecting groups are removed by suitable
deprotection reactions. Where the phosphoramidate employed is
dibenzylphosphoramidate, the benzyl protecting groups may be
removed by hydrogenation of the compound over Pd/C under 1
atmosphere of hydrogen at room temperature. The preparation of
phosphoric acid
mono-[3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)-
-2,5-dioxo-pyrrolidin-1-ylmethyl] ester from
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-1-hydroxymethyl-4-(1H-i-
ndol-3-yl)-pyrrolidine-2,5-dione is described in Example 56, steps
2-3.
[0115] Compounds of formulas III, IIIa, IVa, IVb, Va, or Vb where
R7 is a carboxylic acid group, or an amino carboxylic acid group,
may be prepared by coupling the desired hydroxymethylene derivative
with a carboxylic acid or amino carboxylic acid (amino acid) under
conditions suitable for the formation of an ester linkage. A
variety of dehydrating agents, including DCC
(dicyclohexylcarbodiimide), HBTU
(O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate), or BOP
((benzotriazol-1-yloxy)tris(dimethylamino) phosphonium
hexafluorophosphate) may be employed to drive the formation of the
ester bond. In a preferred embodiment, the reactions are conducted
in anhydrous THF in the presence of HBTU and DIEPA
(N,N-diisopropylethylamine) at room temperature for 10 hours to 24
hours. Following completion of the dehydration reaction, solvent is
removed under reduced pressure and the compounds are taken up in an
organic solvent (e.g., ethyl acetate) and washed with water. The
organic layer is dried and the residue purified by silica gel
chromatography as necessary.
[0116] Where R7 is an amino carboxylic acid group, the starting
materials for introducing the amino carboxylic acid group must
contain a suitably protected amine. A variety of suitable
amine-protecting groups may be advantageously employed including
carbobenzyloxy-protected amines (e.g., the reactions may employ
N-carbobenzyloxy glycine or N-carbobenzyloxy alanine etc.).
Subsequent deprotection will yield the free product. Where the
protecting group employed is carbobenzyloxy, deprotection may be
accomplished by treating the amine protected product suspended in
methanol with HCl (4M) in ethyl acetate in the presence of
palladium on charcoal (Pd/C) under 1 atmosphere of hydrogen for 1-3
hours at room temperature. Examples 58-60 describe the preparation
of compounds where R7 is a carboxylic acid group, or an amino
carboxylic acid group.
[0117] Compounds of formulas III, IIIa, IVa, IVb, Va, or Vb where
R7 is a peptide, may be prepared by coupling the desired
hydroxymethylene derivative with a peptide bearing a free
carboxylic acid group to form an ester linkage. Linking of a
carboxyl functionality of a peptide and the hydroxymethylene group
in an ester linkage may be conducted employing a suitably protected
peptide, bearing for example, protected free amine groups protected
with conventional N-protecting groups.
[0118] Conditions suitable for the formation of an ester linkage,
include those employing dehydrating agents, such as those described
for the preparation of compounds of formulas III, IIIa, IVa, IVb,
Va, or Vb where R4 is --CH.sub.2R7 and R7 is a carboxylic acid
group, or an amino carboxylic acid group.
2.1.5. Preparation of Compounds of Formula III, IIIa, IVa, IVb, Va,
and Vb where R4 is --(C.sub.1-C.sub.5) alkyl
[0119] Compounds of formulas III, IIIa, IVa, IVb, Va, or Vb where
R4 is a --(C.sub.1-C.sub.6) alkyl may be prepared by reacting by
reacting the desired compound of formula III, IIIa, IVa, IVb, Va,
or Vb where R4 is H with a (C.sub.1-C.sub.6) alkyl halide, where
the halide is preferably Cl, Br or I, in the presence of a suitable
base at room temperature. Suitable bases include organic bases such
as potassium tert-butoxide, sodium methoxide, and inorganic bases
such as KOH, NaOH and K.sub.2CO.sub.3. Suitable solvents include
polar aprotic solvents such as DMSO, THF, dioxane or other ethers,
or DMF. In an alternative embodiment, the compound of formula III,
IIIa, IVa, IVb, Va, or Vb where R4 is H is reacted with an organic
or inorganic base to yield the conjugate base of the compound of
formula III, IIIa, IVa, IVb, Va, or Vb, and the conjugate base is
then reacted with the alkylhalide. Where the alkyl group is
introduced into a compound of formula III or IIIa, the resulting
alkylated compounds can be reduced to yield compounds of formulas
IVa and IVb, Va and Vb, or a mixture of compounds of formulas IVa,
IVb, Va, and Vb employing the reduction procedures described in
Section 1(b)(1). Example 61 describes the preparation of
3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-methylindol-3-yl)-1-methyl pyrrole-2, 5-dione
using iodomethane as an alkylating agent, and its reduction by
catalytic hydrogenation to yield
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1-methylindol-3-yl)-1-methyl pyrrolidine-2,
5-dione.
[0120] Compounds of formulas III, IIIa, IVa, IVb, Va, or Vb where
R4 is a --(C.sub.1-C.sub.6) alkyl group may also be prepared by
reacting the desired compound of formula III, IIIa, IVa, IVb, Va,
or Vb where R4 is H with a (C.sub.1-C.sub.6) alkyl alcohol in the
presence of diethylzodicarboxylate (DEAD) and triphenylphosphine.
(See, e.g., Mitsunobu, O.; Wade, M.; Sano, T. J. Am Chem. Soc. 94:
694 (1972); Hughes, D. L., Organic Reactions, 42; 335-656(1992)).
The reactions may be conducted in a variety of solvents including
tetrahydrofuran (THF), dichloromethane, chloroform, acetonitrile,
and benzene, preferably the solvent is THF.
2.1.6. Separation of Compounds of Formula III, IIIa, IVa, IVb, Va,
and Vb
[0121] Where the isolation of an individual product having the
structure of formulas Ill, IIIa, IVa, IVb, Va, or Vb is desired,
the products may be separated by chromatography on one or more
chromatography media. Chromatography may be carried out on a
preparative scale or on an analytical scale to determine the
identity and purity of the products present in a sample. Although
any suitable chromatography media including, but not limited to,
silica, reverse phase, ion exchange, chiral chromatographic media,
or any combination thereof, may be advantageously employed for
separations, the suitability of specific chromatographic media and
conditions for the separation of products having formulas III,
IIIa, IVa, IVb, Va, and Vb will depend upon the substituents
present on the compounds. In preferred embodiments, chromatographic
separations are conducted employing HPLC. In other preferred
embodiments the separation is carried out using supercritical fluid
chromatography. Where supercritical fluid chromatography is
employed, CO.sub.2, or mixtures of CO.sub.2 with other solvents
including acetonitrile (ACN), methanol, ethanol, isopropanol, or
hexane, are the preferred mobile phase, with mixtures of CO.sub.2
and methanol most preferred. A variety of chromatographic media
(stationary phases) may be employed in supercritical fluid
chromatography including, but not limited to: ChiralCel OA, OB, OD,
or OJ; ChiralPak AD or AS; Cyclobond I, II, or III; and Chirobiotic
T, V, and R media.
[0122] In more preferred embodiments, where the products are
individual isomers of formulas IVa, IVb, Va, or Vb, mixtures
containing two or more of the isomeric forms may be separated by
using supercritical fluid chromatography on chiral media. In one
more preferred embodiment, separations are conducted on
CHIRALPAK.RTM. AD columns (Daicel (U.S.A.) Inc. Fort Lee, N.J.). In
that embodiment, products are applied to the AD column in a mixture
of methanol and acetonitrile, or in acetonitrile, and the column is
subsequently eluted with 35% methanol in CO2 (65%). The separation
of 3(R),4(S)-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione and
3(S),4(R)-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione on a
CHIRALPAK.RTM. AD column is set forth in Example 4. The separation
of (+) trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione and (-)
trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione is set forth
in Example 5.
[0123] The individual racemic forms of compounds of formulas III,
IIIa, IVa, IVb, Va, or Va may also be resolved by physical methods,
such as, for example, fractional crystallization or crystallization
of diastereomeric derivatives. In addition, individual optical
isomers can be obtained from racemic mixtures by conventional
methods, such as, for example, salt formation with an optically
active acid, where applicable, followed by crystallization.
2.2. Preparation of Compounds of Formula I and II where Y is a
Bond
[0124] Compounds of formulas I and II, which are employed in the
synthesis of pyrroloquinolinyl-pyrrole-2,5-dione of formulas III
and IIIa, may be purchased or obtained via a variety of synthetic
routes such as those set forth below.
2.2.1. Preparation of Compounds of Formula I where Y is a Bond
[0125] Compounds of formula I may be prepared from the
corresponding compound of formula A, where X is selected from the
group consisting of --(CH.sub.2)--, --(NR8)-, S and O, R8 is
selected from the group consisting of hydrogen, --(C.sub.1-C.sub.6)
alkyl, --(C.sub.1-C.sub.6) substituted alkyl, --(C.sub.3-C.sub.9)
cycloalkyl, --(C.sub.3-C.sub.9) substituted cycloalkyl, and
--O--(C.sub.1-C.sub.6) alkyl, and m is 1 or 2. Exemplary compounds
of formula A include 1,2,3,4-tetra hydroquinoline,
1,2,3,4-tetrahydro-quinoxaline, 3,4-dihydro-2H-benzo[1,4]oxazine,
3,4-dihydro-2H-benzo[1,4]thiazine,
2,3,4,5-tetrahydro-1H-benzo[b]azepine,
2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepine,
6,7,8,9-tetrahydro-5-oxa-9-aza-benzocycloheptene, or
2,3,4,5-tetrahydro-benzo[b][1,4]thiazepine). The preparation begins
with the conversion of a compound of formula A to the corresponding
3-substituted-2-oxopropionic acid ethyl ester of formula B. The
ethyl ester of formula B is cyclized to form a compound of formula
C, which is converted to the free acid D, which is decarboxylated
to yield the desired tricyclic product E. Subsequent reaction of
the tricyclic product E with oxalyl chloride and work-up in
alcoholic base yields the corresponding compound of formula I.
Scheme 3 illustrates the reaction sequence beginning with compounds
of formula A, which is further illustrated in Example 1, steps 1-5
for the preparation of 5, 6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl) oxoacetic methyl ester of formula I from
1,2,3,4-tetrahydroquinoline and bromoethylpyrruvate
(3-bromo-pyruvic acid ethyl ester).
[0126] Some suitable conditions for the conversion of compounds of
formula A into compounds of formula I through the reaction sequence
of Scheme 3 are described herein. Compounds of formula A may be
converted to the corresponding 3-substituted-2-oxopropionic acid
ethyl ester of formula B by treatment with bromoethyl pyruvate in
an anhydrous ether, such as THF, at room temperature for about 24
hours. Treatment of the 3-substituted-2-oxopropionic acid ethyl
ester of formula B with anhydrous MgCl in 2-methoxyethanol at about
125.degree. C. for 30 minutes to 2 hours, preferably for 1 hour,
results in the formation of the corresponding tricyclic carboxylic
acid ester of formula C. Subsequent conversion of this compound to
the free acid of formula D may be accomplished by hydrolysis in
aqueous base. In preferred embodiments the reaction is carried out
in an aqueous base, including but not limited to NaOH or KOH, in
the presence of alcohol as a co-solvent. Preferred alcohol
co-solvents include methanol, ethanol, n-propanol, and isopropanol,
with ethanol as a more preferred co-solvent. Reactions are
typically conducted by heating the mixture to reflux for 2 hours,
although the time and temperature of the reaction may be varied as
needed. Oxidative decarboxylation of compounds of formula D may be
conducted by a variety of procedures suitable for the
decarboxylation of aromatic acids. In preferred embodiments the
decarboxylation of compounds of formula D is conducted by heating
the free acid with copper-chromite (CuO--Cr.sub.2O.sub.3) in
quinolone for about 2 hours to yield the decarboxylated product of
formula E. Conversion of compounds of formula E to compounds of
formula I may be accomplished by reaction with oxalyl chloride,
followed by treatment with a mixture of an anhydrous alcohol and
the alkaline metal salt of the alcohol, preferably sodium
methoxide, or sodium ethoxide. The reaction of oxalyl chloride with
compounds of formula E is typically conducted in anhydrous polar
aprotic solvents including ethers at a temperature from about
-78.degree. C. to about 10.degree. C. In preferred embodiments, the
reaction is conducted at a temperature from about -25.degree. C. to
about 5.degree. C. employing an ether as a solvent. In more
preferred embodiments the reaction is conducted at 0.degree. C.
Preferred solvents for conducting the reaction include, but are not
limited to tetrahydrofuran (THF), tetrahydropyran, diethyl ether
and the like.
##STR00007##
2.2.2. Preparation of Compounds of Formula II
[0127] Compounds of formula II, which are substituted acetamides,
may be purchased or prepared from commercially available starting
materials. Commercially available acetamides including:
indole-3-acetamide, 2-(5-methyl-1H-indol-3-yl)acetamide,
2-(5-methoxy-1H-indol-3-yl)acetamide,
2-(4-hydroxy-1H-indol-3-yl)acetamide, 2-phenylacetamide,
2-(4-methylphenyl)acetamide, 4-hydroxyphenylacetamide,
4-hydroxyphenylacetamide,
N-cyclopentyl-2-(4-hydroxy-2-oxo-1,2-dihydro-3-quinolinyl)acetamide,
2-phenoxyacetamide, 2-(2-methylphenoxy)acetamide,
2-(4-fluorophenoxy)acetamide, 2-(4-pyridinyl)acetamide, and
2-[(4-chlorophenyl)sulfanyl] acetamide are available from a variety
of sources including Sigma Aldrich Chemical Co., St. Louis Mo. A
compound of formula II may also be prepared from its corresponding
free acid by conversion of the free acid to its acid chloride
followed by reaction with ammonia.
2.3. Additional Routes for the Preparation of
Pyrroloquinolinyl-Pyrrolidine-2,5-Diones
[0128] In addition to those routes for the preparation of
pyrroloquinolinyl-pyrrolidine-2,5-diones described above,
additional routes of preparing the compounds exemplified for
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione are
described in Examples 62-64.
3. Methods of Treatment
[0129] As used herein, a "subject" can be any mammal, e.g., a
human, a primate, mouse, rat, dog, cat, cow, horse, pig, sheep,
goat, camel. In a preferred aspect, the subject is a human.
[0130] As used herein, a "subject in need thereof" is a subject
having a cell proliferative disorder, or a subject having an
increased risk of developing a cell proliferative disorder relative
to the population at large. In one aspect, a subject in need
thereof has a precancerous condition. In a preferred aspect, a
subject in need thereof has cancer.
[0131] As used herein, the term "cell proliferative disorder"
refers to conditions in which unregulated or abnormal growth, or
both, of cells can lead to the development of an unwanted condition
or disease, which may or may not be cancerous. In one aspect, a
cell proliferative disorder includes a non-cancerous condition,
e.g., rheumatoid arthritis; inflammation; autoimmune disease;
lymphoproliferative conditions; acromegaly; rheumatoid spondylitis;
osteoarthritis; gout, other arthritic conditions; sepsis; septic
shock; endotoxic shock; gram-negative sepsis; toxic shock syndrome;
asthma; adult respiratory distress syndrome; chronic obstructive
pulmonary disease; chronic pulmonary inflammation; inflammatory
bowel disease; Crohn's disease; psoriasis; eczema; ulcerative
colitis; pancreatic fibrosis; hepatic fibrosis; acute and chronic
renal disease; irritable bowel syndrome; pyresis; restenosis;
cerebral malaria; stroke and ischemic injury; neural trauma;
Alzheimer's disease; Huntington's disease; Parkinson's disease;
acute and chronic pain; allergic rhinitis; allergic conjunctivitis;
chronic heart failure; acute coronary syndrome; cachexia; malaria;
leprosy; leishmaniasis; Lyme disease; Reiter's syndrome; acute
synovitis; muscle degeneration, bursitis; tendonitis;
tenosynovitis; herniated, ruptures, or prolapsed intervertebral
disk syndrome; osteopetrosis; thrombosis; restenosis; silicosis;
pulmonary sarcosis; bone resorption diseases, such as osteoporosis;
graft-versus-host reaction; Multiple Sclerosis; lupus;
fibromyalgia; AIDS and other viral diseases such as Herpes Zoster,
Herpes Simplex I or II, influenza virus and cytomegalovirus; and
diabetes mellitus. In another aspect, a cell proliferative disorder
includes a precancer or a precancerous condition. In another
aspect, a cell proliferative disorder includes cancer. Various
cancers to be treated include but are not limited to breast cancer,
lung cancer, colorectal cancer, pancreatic cancer, ovarian cancer,
prostate cancer, renal carcinoma, hepatoma, brain cancer, melanoma,
multiple myeloma, chronic myelogenous leukemia, hematologic tumor,
and lymphoid tumor, including metastatic lesions in other tissues
or organs distant from the primary tumor site.
[0132] Cancers to be treated include but are not limited to
sarcoma, carcinoma, and adenocarcinoma. In one aspect, a "precancer
cell" or "precancerous cell" is a cell manifesting a cell
proliferative disorder that is a precancer or a precancerous
condition. In another aspect, a "cancer cell" or "cancerous cell"
is a cell manifesting a cell proliferative disorder that is a
cancer. Any reproducible means of measurement may be used to
identify cancer cells or precancerous cells. In a preferred aspect,
cancer cells or precancerous cells are identified by histological
typing or grading of a tissue sample (e.g., a biopsy sample). In
another aspect, cancer cells or precancerous cells are identified
through the use of appropriate molecular markers.
[0133] A "cell proliferative disorder of the hematologic system" is
a cell proliferative disorder involving cells of the hematologic
system. In one aspect, a cell proliferative disorder of the
hematologic system includes lymphoma, leukemia, myeloid neoplasms,
mast cell neoplasms, myelodysplasia, benign monoclonal gammopathy,
lymphomatoid granulomatosis, lymphomatoid papulosis, polycythemia
vera, chronic myelocytic leukemia, agnogenic myeloid metaplasia,
and essential thrombocythemia. In another aspect, a cell
proliferative disorder of the hematologic system includes
hyperplasia, dysplasia, and metaplasia of cells of the hematologic
system. In a preferred aspect, compositions of the present
invention may be used to treat a cancer selected from the group
consisting of a hematologic cancer of the present invention or a
hematologic cell proliferative disorder of the present invention.
In one aspect, a hematologic cancer of the present invention
includes multiple myeloma, lymphoma (including Hodgkin's lymphoma,
non-Hodgkin's lymphoma, childhood lymphomas, and lymphomas of
lymphocytic and cutaneous origin), leukemia (including childhood
leukemia, hairy-cell leukemia, acute lymphocytic leukemia, acute
myelocytic leukemia, chronic lymphocytic leukemia, chronic
myelocytic leukemia, chronic myelogenous leukemia, and mast cell
leukemia), myeloid neoplasms and mast cell neoplasms.
[0134] A "cell proliferative disorder of the lung" is a cell
proliferative disorder involving cells of the lung. In one aspect,
cell proliferative disorders of the lung include all forms of cell
proliferative disorders affecting lung cells. In one aspect, cell
proliferative disorders of the lung include lung cancer, a
precancer or precancerous condition of the lung, benign growths or
lesions of the lung, and malignant growths or lesions of the lung,
and metastatic lesions in tissue and organs in the body other than
the lung. In a preferred aspect, compositions of the present
invention may be used to treat lung cancer or cell proliferative
disorders of the lung. In one aspect, lung cancer includes all
forms of cancer of the lung. In another aspect, lung cancer
includes malignant lung neoplasms, carcinoma in situ, typical
carcinoid tumors, and atypical carcinoid tumors. In another aspect,
lung cancer includes small cell lung cancer ("SCLC"), non-small
cell lung cancer ("NSCLC"), squamous cell carcinoma,
adenocarcinoma, small cell carcinoma, large cell carcinoma,
adenosquamous cell carcinoma, and mesothelioma. In another aspect,
lung cancer includes "scar carcinoma," bronchioalveolar carcinoma,
giant cell carcinoma, spindle cell carcinoma, and large cell
neuroendocrine carcinoma. In another aspect, lung cancer includes
lung neoplasms having histologic and ultrastructual heterogeneity
(e.g., mixed cell types).
[0135] In one aspect, cell proliferative disorders of the lung
include all forms of cell proliferative disorders affecting lung
cells. In one aspect, cell proliferative disorders of the lung
include lung cancer, precancerous conditions of the lung. In one
aspect, cell proliferative disorders of the lung include
hyperplasia, metaplasia, and dysplasia of the lung. In another
aspect, cell proliferative disorders of the lung include
asbestos-induced hyperplasia, squamous metaplasia, and benign
reactive mesothelial metaplasia. In another aspect, cell
proliferative disorders of the lung include replacement of columnar
epithelium with stratified squamous epithelium, and mucosal
dysplasia. In another aspect, individuals exposed to inhaled
injurious environmental agents such as cigarette smoke and asbestos
may be at increased risk for developing cell proliferative
disorders of the lung. In another aspect, prior lung diseases that
may predispose individuals to development of cell proliferative
disorders of the lung include chronic interstitial lung disease,
necrotizing pulmonary disease, scleroderma, rheumatoid disease,
sarcoidosis, interstitial pneumonitis, tuberculosis, repeated
pneumonias, idiopathic pulmonary fibrosis, granulomata, asbestosis,
fibrosing alveolitis, and Hodgkin's disease.
[0136] A "cell proliferative disorder of the colon" is a cell
proliferative disorder involving cells of the colon. In a preferred
aspect, the cell proliferative disorder of the colon is colon
cancer. In a preferred aspect, compositions of the present
invention may be used to treat colon cancer or cell proliferative
disorders of the colon. In one aspect, colon cancer includes all
forms of cancer of the colon. In another aspect, colon cancer
includes sporadic and hereditary colon cancers. In another aspect,
colon cancer includes malignant colon neoplasms, carcinoma in situ,
typical carcinoid tumors, and atypical carcinoid tumors. In another
aspect, colon cancer includes adenocarcinoma, squamous cell
carcinoma, and adenosquamous cell carcinoma. In another aspect,
colon cancer is associated with a hereditary syndrome selected from
the group consisting of hereditary nonpolyposis colorectal cancer,
familial adenomatous polyposis, Gardner's syndrome, Peutz-Jeghers
syndrome, Turcot's syndrome and juvenile polyposis. In another
aspect, colon cancer is caused by a hereditary syndrome selected
from the group consisting of hereditary nonpolyposis colorectal
cancer, familial adenomatous polyposis, Gardner's syndrome,
Peutz-Jeghers syndrome, Turcot's syndrome and juvenile
polyposis.
[0137] In one aspect, cell proliferative disorders of the colon
include all forms of cell proliferative disorders affecting colon
cells. In one aspect, cell proliferative disorders of the colon
include colon cancer, precancerous conditions of the colon,
adenomatous polyps of the colon and metachronous lesions of the
colon. In one aspect, a cell proliferative disorder of the colon
includes adenoma. In one aspect, cell proliferative disorders of
the colon are characterized by hyperplasia, metaplasia, and
dysplasia of the colon. In another aspect, prior colon diseases
that may predispose individuals to development of cell
proliferative disorders of the colon include prior colon cancer. In
another aspect, current disease that may predispose individuals to
development of cell proliferative disorders of the colon include
Crohn's disease and ulcerative colitis. In one aspect, a cell
proliferative disorder of the colon is associated with a mutation
in a gene selected from the group consisting of p53, rus, FAP and
DCC. In another aspect, an individual has an elevated risk of
developing a cell proliferative disorder of the colon due to the
presence of a mutation in a gene selected from the group consisting
of p53, ras, FAP and DCC.
[0138] A "cell proliferative disorder of the prostate" is a cell
proliferative disorder involving cells of the prostate. In one
aspect, cell proliferative disorders of the prostate include all
forms of cell proliferative disorders affecting prostate cells. In
one aspect, cell proliferative disorders of the prostate include
prostate cancer, a precancer or precancerous condition of the
prostate, benign growths or lesions of the prostate, and malignant
growths or lesions of the prostate, and metastatic lesions in
tissue and organs in the body other than the prostate. In another
aspect, cell proliferative disorders of the prostate include
hyperplasia, metaplasia, and dysplasia of the prostate.
[0139] A "cell proliferative disorder of the skin" is a cell
proliferative disorder involving cells of the skin. In one aspect,
cell proliferative disorders of the skin include all forms of cell
proliferative disorders affecting skin cells. In one aspect, cell
proliferative disorders of the skin include a precancer or
precancerous condition of the skin, benign growths or lesions of
the skin, melanoma, malignant melanoma and other malignant growths
or lesions of the skin, and metastatic lesions in tissue and organs
in the body other than the skin. In another aspect, cell
proliferative disorders of the skin include hyperplasia,
metaplasia, and dysplasia of the skin.
[0140] A "cell proliferative disorder of the ovary" is a cell
proliferative disorder involving cells of the ovary. In one aspect,
cell proliferative disorders of the ovary include all forms of cell
proliferative disorders affecting cells of the ovary. In one
aspect, cell proliferative disorders of the ovary include a
precancer or precancerous condition of the ovary, benign growths or
lesions of the ovary, ovarian cancer, malignant growths or lesions
of the ovary, and metastatic lesions in tissue and organs in the
body other than the ovary. In another aspect, cell proliferative
disorders of the skin include hyperplasia, metaplasia, and
dysplasia of cells of the ovary.
[0141] A "cell proliferative disorder of the breast" is a cell
proliferative disorder involving cells of the breast. In one
aspect, cell proliferative disorders of the breast include all
forms of cell proliferative disorders affecting breast cells. In
one aspect, cell proliferative disorders of the breast include
breast cancer, a precancer or precancerous condition of the breast,
benign growths or lesions of the breast, and malignant growths or
lesions of the breast, and metastatic lesions in tissue and organs
in the body other than the breast. In another aspect, cell
proliferative disorders of the breast include hyperplasia,
metaplasia, and dysplasia of the breast.
[0142] In one aspect, a cell proliferative disorder of the breast
is a precancerous condition of the breast. In one aspect,
compositions of the present invention may be used to treat a
precancerous condition of the breast. In one aspect, a precancerous
condition of the breast includes atypical hyperplasia of the
breast, ductal carcinoma in situ (DCIS), intraductal carcinoma,
lobular carcinoma in situ (LCIS), lobular neoplasia, and stage 0 or
grade 0 growth or lesion of the breast (e.g., stage 0 or grade 0
breast cancer, or carcinoma in situ). In another aspect, a
precancerous condition of the breast has been staged according to
the TNM classification scheme as accepted by the American Joint
Committee on Cancer (AJCC), where the primary tumor (T) has been
assigned a stage of T0 or Tis; and where the regional lymph nodes
(N) have been assigned a stage of N0; and where distant metastasis
(M) has been assigned a stage of M0.
[0143] In a preferred aspect, the cell proliferative disorder of
the breast is breast cancer. In a preferred aspect, compositions of
the present invention may be used to treat breast cancer. In one
aspect, breast cancer includes all forms of cancer of the breast.
In one aspect, breast cancer includes primary epithelial breast
cancers. In another aspect, breast cancer includes cancers in which
the breast is involved by other tumors such as lymphoma, sarcoma or
melanoma. In another aspect, breast cancer includes carcinoma of
the breast, ductal carcinoma of the breast, lobular carcinoma of
the breast, undifferentiated carcinoma of the breast, cystosarcoma
phyllodes of the breast, angiosarcoma of the breast, and primary
lymphoma of the breast. In one aspect, breast cancer includes Stage
I, II, IIIA, IIIB, IIIC and IV breast cancer. In one aspect, ductal
carcinoma of the breast includes invasive carcinoma, invasive
carcinoma in situ with predominant intraductal component,
inflammatory breast cancer, and a ductal carcinoma of the breast
with a histologic type selected from the group consisting of
comedo, mucinous (colloid), medullary, medullary with lymphcytic
infiltrate, papillary, scirrhous, and tubular. In one aspect,
lobular carcinoma of the breast includes invasive lobular carcinoma
with predominant in situ component, invasive lobular carcinoma, and
infiltrating lobular carcinoma. In one aspect, breast cancer
includes Paget's disease, Paget's disease with intraductal
carcinoma, and Paget's disease with invasive ductal carcinoma. In
another aspect, breast cancer includes breast neoplasms having
histologic and ultrastructual heterogeneity (e.g., mixed cell
types).
[0144] In a preferred aspect, a compound of the present invention
may be used to treat breast cancer. In one aspect, a breast cancer
that is to be treated includes familial breast cancer. In another
aspect, a breast cancer that is to be treated includes sporadic
breast cancer. In one aspect, a breast cancer that is to be treated
has arisen in a male subject. In one aspect, a breast cancer that
is to be treated has arisen in a female subject. In one aspect, a
breast cancer that is to be treated has arisen in a premenopausal
female subject or a postmenopausal female subject. In one aspect, a
breast cancer that is to be treated has arisen in a subject equal
to or older than 30 years old, or a subject younger than 30 years
old. In one aspect, a breast cancer that is to be treated has
arisen in a subject equal to or older than 50 years old, or a
subject younger than 50 years old. In one aspect, a breast cancer
that is to be treated has arisen in a subject equal to or older
than 70 years old, or a subject younger than 70 years old.
[0145] In one aspect, a breast cancer that is to be treated has
been typed to identify a familial or spontaneous mutation in BRCA1,
BRCA2, or p53. In one aspect, a breast cancer that is to be treated
has been typed as having a HER2/neu gene amplification, as
overexpressing HER2/neu, or as having a low, intermediate or high
level of HER2/neu expression. In another aspect, a breast cancer
that is to be treated has been typed for a marker selected from the
group consisting of estrogen receptor (ER), progesterone receptor
(PR), human epidermal growth factor receptor-2, Ki-67, CA 15-3, CA
27-29, and c-Met. In one aspect, a breast cancer that is to be
treated has been typed as ER-unknown, ER-rich or ER-poor. In
another aspect, a breast cancer that is to be treated has been
typed as ER-negative or ER-positive. ER-typing of a breast cancer
may be performed by any reproducible means. In a preferred aspect,
ER-typing of a breast cancer may be performed as set forth in
Onkologie 27: 175-179 (2004). In one aspect, a breast cancer that
is to be treated has been typed as PR-unknown, PR-rich or PR-poor.
In another aspect, a breast cancer that is to be treated has been
typed as PR-negative or PR-positive. In another aspect, a breast
cancer that is to be treated has been typed as receptor positive or
receptor negative. In one aspect, a breast cancer that is to be
treated has been typed as being associated with elevated blood
levels of CA 15-3, or CA 27-29, or both.
[0146] In one aspect, a breast cancer that is to be treated
includes a localized tumor of the breast. In one aspect, a breast
cancer that is to be treated includes a tumor of the breast that is
associated with a negative sentinel lymph node (SLN) biopsy. In one
aspect, a breast cancer that is to be treated includes a tumor of
the breast that is associated with a positive sentinel lymph node
(SLN) biopsy. In another aspect, a breast cancer that is to be
treated includes a tumor of the breast that is associated with one
or more positive axillary lymph nodes, where the axillary lymph
nodes have been staged by any applicable method. In another aspect,
a breast cancer that is to be treated includes a tumor of the
breast that has been typed as having nodal negative status (e.g.,
node-negative) or nodal positive status (e.g., node-positive). In
another aspect, a breast cancer that is to be treated includes a
tumor of the breast that has metastasized to other locations in the
body. In one aspect, a breast cancer that is to be treated is
classified as having metastasized to a location selected from the
group consisting of bone, lung, liver, or brain. In another aspect
a breast cancer that is to be treated is classified according to a
characteristic selected from the group consisting of metastatic,
localized, regional, local-regional, locally advanced, distant,
multicentric, bilateral, ipsilateral, contralateral, newly
diagnosed, recurrent, and inoperable.
[0147] In one aspect, a compound of the present invention may be
used to treat or prevent a cell proliferative disorder of the
breast, or to treat or prevent breast cancer, in a subject having
an increased risk of developing breast cancer relative to the
population at large. In one aspect, a subject with an increased
risk of developing breast cancer relative to the population at
large is a female subject with a family history or personal history
of breast cancer. In another aspect, a subject with an increased
risk of developing breast cancer relative to the population at
large is a female subject having a germ-line or spontaneous
mutation in BRCA1 or BRCA2, or both. In one aspect, a subject with
an increased risk of developing breast cancer relative to the
population at large is a female subject with a family history of
breast cancer and a germ-line or spontaneous mutation in BRCA1 or
BRCA2, or both. In another aspect, a subject with an increased risk
of developing breast cancer relative to the population at large is
a female who is greater than 30 years old, greater than 40 years
old, greater than 50 years old, greater than 60 years old, greater
than 70 years old, greater than 80 years old, or greater than 90
years old. In one aspect, a subject with an increased risk of
developing breast cancer relative to the population at large is a
subject with atypical hyperplasia of the breast, ductal carcinoma
in situ (DCIS), intraductal carcinoma, lobular carcinoma in situ
(LCIS), lobular neoplasia, or a stage 0 growth or lesion of the
breast (e.g., stage 0 or grade 0 breast cancer, or carcinoma in
situ).
[0148] In another aspect, a breast cancer that is to be treated has
been histologically graded according to the Scarff-Bloom-Richardson
system, wherein a breast tumor has been assigned a mitosis count
score of 1, 2, or 3; a nuclear pleiomorphism score of 1, 2, or 3; a
tubule formation score of 1, 2, or 3; and a total
Scarff-Bloom-Richardson score of between 3 and 9. In another
aspect, a breast cancer that is to be treated has been assigned a
tumor grade according to the International Consensus Panel on the
Treatment of Breast Cancer selected from the group consisting of
grade 1, grade 1-2, grade 2, grade 2-3, or grade 3.
[0149] In one aspect, a cancer that is to be treated has been
staged according to the American Joint Committee on Cancer (AJCC)
TNM classification system, where the tumor (T) has been assigned a
stage of TX, T1, T1mic, T1a, T1b, T1c, T2, T3, T4, T4a, T4b, T4c,
or T4d; and where the regional lymph nodes (N) have been assigned a
stage of NX, N0, N1, N2, N2a, N2b, N3, N3a, N3b, or N3c; and where
distant metastasis (M) has been assigned a stage of MX, M0, or M1.
In another aspect, a cancer that is to be treated has been staged
according to an American Joint Committee on Cancer (AJCC)
classification as Stage I, Stage IIA, Stage IIB, Stage IIIA, Stage
IIIB, Stage IIIC, or Stage IV.
[0150] In another aspect, a cancer that is to be treated has been
assigned a grade according to an AJCC classification as Grade GX
(e.g., grade cannot be assessed), Grade 1, Grade 2, Grade 3 or
Grade 4. In another aspect, a cancer that is to be treated has been
staged according to an AJCC pathologic classification (pN) of pNX,
pN0, PN0 (I-), PN0 (I+), PN0 (mol-), PN0 (mol+), PN1, PN1(mi),
PN1a, PN1 b, PN1c, pN2, pN2a, pN2b, pN3, pN3a, pN3b, or pN3c.
[0151] In one aspect, a cancer that is to be treated includes a
tumor that has been determined to be less than or equal to about 2
centimeters in diameter. In another aspect, a cancer that is to be
treated includes a tumor that has been determined to be from about
2 to about 5 centimeters in diameter. In another aspect, a cancer
that is to be treated includes a tumor that has been determined to
be greater than or equal to about 3 centimeters in diameter. In
another aspect, a cancer that is to be treated includes a tumor
that has been determined to be greater than 5 centimeters in
diameter. In another aspect, a cancer that is to be treated is
classified by microscopic appearance as well differentiated,
moderately differentiated, poorly differentiated, or
undifferentiated. In another aspect, a cancer that is to be treated
is classified by microscopic appearance with respect to mitosis
count (e.g., amount of cell division) or nuclear pleiomorphism
(e.g., change in cells). In another aspect, a cancer that is to be
treated is classified by microscopic appearance as being associated
with areas of necrosis (e.g., areas of dying or degenerating
cells). In one aspect, a cancer that is to be treated is classified
as having an abnormal karyotype, having an abnormal number of
chromosomes, or having one or more chromosomes that are abnormal in
appearance. In one aspect, a cancer that is to be treated is
classified as being aneuploid, triploid, tetraploid, or as having
an altered ploidy. In one aspect, a cancer that is to be treated is
classified as having a chromosomal translocation, or a deletion or
duplication of an entire chromosome, or a region of deletion,
duplication or amplification of a portion of a chromosome.
[0152] In one aspect, a cancer that is to be treated is evaluated
by DNA cytometry, flow cytometry, or image cytometry. In one
aspect, a cancer that is to be treated has been typed as having
10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of cells in the
synthesis stage of cell division (e.g., in S phase of cell
division). In one aspect, a cancer that is to be treated has been
typed as having a low S-phase fraction or a high S-phase
fraction.
[0153] As used herein, a "normal cell" is a cell that cannot be
classified as part of a "cell proliferative disorder." In one
aspect, a normal cell lacks unregulated or abnormal growth, or
both, that can lead to the development of an unwanted condition or
disease. Preferably, a normal cell possesses normally functioning
cell cycle checkpoint control mechanisms.
[0154] As used herein, "contacting a cell" refers to a condition in
which a compound or other composition of matter is in direct
contact with a cell, or is close enough to induce a desired
biological effect in a cell.
[0155] As used herein, "candidate compound" refers to a compound of
the present invention that has been or will be tested in one or
more in vitro or in vivo biological assays, in order to determine
if that compound is likely to elicit a desired biological or
medical response in a cell, tissue, system, animal or human that is
being sought by a researcher or clinician. In one aspect, a
candidate compound is a compound of formula IIIa; in another
aspect, a candidate compound is a compound of formula IVa, IVb, Va,
or Vb. In a preferred aspect, the biological or medical response is
treatment of cancer. In another aspect, the biological or medical
response is treatment or prevention of a cell proliferative
disorder. In one aspect, in vitro or in vivo biological assays
include, but are not limited to, enzymatic activity assays,
electrophoretic mobility shift assays, reporter gene assays, in
vitro cell viability assays, and the assays set forth in Examples
65-73 herein.
[0156] As used herein, "monotherapy" refers to administration of a
single active or therapeutic compound to a subject in need thereof.
Preferably, monotherapy will involve administration of a
therapeutically effective amount of an active compound. For
example, cancer monotherapy with
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione comprises
administration of a therapeutically effective amount of
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or
derivative thereof, to a subject in need of treatment of cancer.
Monotherapy may be contrasted with combination therapy, in which a
combination of multiple active compounds is administered,
preferably with each component of the combination present in a
therapeutically effective amount. In one aspect, monotherapy with a
compound of the present invention is more effective than
combination therapy in inducing a desired biological effect.
[0157] As used herein, "treating" describes the management and care
of a patient for the purpose of combating a disease, condition, or
disorder and includes the administration of a compound of the
present invention to prevent the onset of the symptoms or
complications, alleviating the symptoms or complications, or
eliminating the disease, condition or disorder.
[0158] In one aspect, treating cancer results in a reduction in
size of a tumor. A reduction in size of a tumor may also be
referred to as "tumor regression." Preferably, after treatment,
tumor size is reduced by 5% or greater relative to its size prior
to treatment; more preferably, tumor size is reduced by 10% or
greater; more preferably, reduced by 20% or greater; more
preferably, reduced by 30% or greater; more preferably, reduced by
40% or greater; even more preferably, reduced by 50% or greater;
and most preferably, reduced by greater than 75% or greater. Size
of a tumor may be measured by any reproducible means of
measurement. In a preferred aspect, size of a tumor may be measured
as a diameter of the tumor.
[0159] In another aspect, treating cancer results in a reduction in
tumor volume. Preferably, after treatment, tumor volume is reduced
by 5% or greater relative to its size prior to treatment; more
preferably, tumor volume is reduced by 10% or greater; more
preferably, reduced by 20% or greater; more preferably, reduced by
30% or greater; more preferably, reduced by 40% or greater; even
more preferably, reduced by 50% or greater; and most preferably,
reduced by greater than 75% or greater. Tumor volume may be
measured by any reproducible means of measurement.
[0160] In another aspect, treating cancer results in a decrease in
number of tumors. Preferably, after treatment, tumor number is
reduced by 5% or greater relative to number prior to treatment;
more preferably, tumor number is reduced by 10% or greater; more
preferably, reduced by 20% or greater; more preferably, reduced by
30% or greater; more preferably, reduced by 40% or greater; even
more preferably, reduced by 50% or greater; and most preferably,
reduced by greater than 75%. Number of tumors may be measured by
any reproducible means of measurement. In a preferred aspect,
number of tumors may be measured by counting tumors visible to the
naked eye or at a specified magnification. In a preferred aspect,
the specified magnification is 2.times., 3.times., 4.times.,
5.times., 10.times., or 50.times..
[0161] In another aspect, treating cancer results in a decrease in
number of metastatic lesions in other tissues or organs distant
from the primary tumor site. Preferably, after treatment, the
number of metastatic lesions is reduced by 5% or greater relative
to number prior to treatment; more preferably, the number of
metastatic lesions is reduced by 10% or greater; more preferably,
reduced by 20% or greater; more preferably, reduced by 30% or
greater; more preferably, reduced by 40% or greater; even more
preferably, reduced by 50% or greater; and most preferably, reduced
by greater than 75%. The number of metastatic lesions may be
measured by any reproducible means of measurement. In a preferred
aspect, the number of metastatic lesions may be measured by
counting metastatic lesions visible to the naked eye or at a
specified magnification. In a preferred aspect, the specified
magnification is 2.times., 3.times., 4.times., 5.times., 10.times.,
or 50.times..
[0162] In another aspect, treating cancer results in an increase in
average survival time of a population of treated subjects in
comparison to a population receiving carrier alone. Preferably, the
average survival time is increased by more than 30 days; more
preferably, by more than 60 days; more preferably, by more than 90
days; and most preferably, by more than 120 days. An increase in
average survival time of a population may be measured by any
reproducible means. In a preferred aspect, an increase in average
survival time of a population may be measured, for example, by
calculating for a population the average length of survival
following initiation of treatment with an active compound. In
another preferred aspect, an increase in average survival time of a
population may also be measured, for example, by calculating for a
population the average length of survival following completion of a
first round of treatment with an active compound.
[0163] In another aspect, treating cancer results in an increase in
average survival time of a population of treated subjects in
comparison to a population of untreated subjects. Preferably, the
average survival time is increased by more than 30 days; more
preferably, by more than 60 days; more preferably, by more than 90
days; and most preferably, by more than 120 days. An increase in
average survival time of a population may be measured by any
reproducible means. In a preferred aspect, an increase in average
survival time of a population may be measured, for example, by
calculating for a population the average length of survival
following initiation of treatment with an active compound. In
another preferred aspect, an increase in average survival time of a
population may also be measured, for example, by calculating for a
population the average length of survival following completion of a
first round of treatment with an active compound.
[0164] In another aspect, treating cancer results in increase in
average survival time of a population of treated subjects in
comparison to a population receiving monotherapy with a drug that
is not a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, analog or derivative thereof.
Preferably, the average survival time is increased by more than 30
days; more preferably, by more than 60 days; more preferably, by
more than 90 days; and most preferably, by more than 120 days. An
increase in average survival time of a population may be measured
by any reproducible means. In a preferred aspect, an increase in
average survival time of a population may be measured, for example,
by calculating for a population the average length of survival
following initiation of treatment with an active compound. In
another preferred aspect, an increase in average survival time of a
population may also be measured, for example, by calculating for a
population the average length of survival following completion of a
first round of treatment with an active compound.
[0165] In another aspect, treating cancer results in a decrease in
the mortality rate of a population of treated subjects in
comparison to a population receiving carrier alone. In another
aspect, treating cancer results in a decrease in the mortality rate
of a population of treated subjects in comparison to an untreated
population. In a further aspect, treating cancer results a decrease
in the mortality rate of a population of treated subjects in
comparison to a population receiving monotherapy with a drug that
is not a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, analog or derivative thereof.
Preferably, the mortality rate is decreased by more than 2%; more
preferably, by more than 5%; more preferably, by more than 10%; and
most preferably, by more than 25%. In a preferred aspect, a
decrease in the mortality rate of a population of treated subjects
may be measured by any reproducible means. In another preferred
aspect, a decrease in the mortality rate of a population may be
measured, for example, by calculating for a population the average
number of disease-related deaths per unit time following initiation
of treatment with an active compound. In another preferred aspect,
a decrease in the mortality rate of a population may also be
measured, for example, by calculating for a population the average
number of disease-related deaths per unit time following completion
of a first round of treatment with an active compound.
[0166] In another aspect, treating cancer results in a decrease in
tumor growth rate. Preferably, after treatment, tumor growth rate
is reduced by at least 5% relative to number prior to treatment;
more preferably, tumor growth rate is reduced by at least 10%; more
preferably, reduced by at least 20%; more preferably, reduced by at
least 30%; more preferably, reduced by at least 40%; more
preferably, reduced by at least 50%; even more preferably, reduced
by at least 50%; and most preferably, reduced by at least 75%.
Tumor growth rate may be measured by any reproducible means of
measurement. In a preferred aspect, tumor growth rate is measured
according to a change in tumor diameter per unit time.
[0167] In another aspect, treating cancer results in a decrease in
tumor regrowth. Preferably, after treatment, tumor regrowth is less
than 5%; more preferably, tumor regrowth is less than 10%; more
preferably, less than 20%; more preferably, less than 30%; more
preferably, less than 40%; more preferably, less than 50%; even
more preferably, less than 50%; and most preferably, less than 75%.
Tumor regrowth may be measured by any reproducible means of
measurement. In a preferred aspect, tumor regrowth is measured, for
example, by measuring an increase in the diameter of a tumor after
a prior tumor shrinkage that followed treatment. In another
preferred aspect, a decrease in tumor regrowth is indicated by
failure of tumors to reoccur after treatment has stopped.
[0168] In another aspect, treating or preventing a cell
proliferative disorder results in a reduction in the rate of
cellular proliferation. Preferably, after treatment, the rate of
cellular proliferation is reduced by at least 5%; more preferably,
by at least 10%; more preferably, by at least 20%; more preferably,
by at least 30%; more preferably, by at least 40%; more preferably,
by at least 50%; even more preferably, by at least 50%; and most
preferably, by at least 75%. The rate of cellular proliferation may
be measured by any reproducible means of measurement. In a
preferred aspect, the rate of cellular proliferation is measured,
for example, by measuring the number of dividing cells in a tissue
sample per unit time.
[0169] In another aspect, treating or preventing a cell
proliferative disorder results in a reduction in the proportion of
proliferating cells. Preferably, after treatment, the proportion of
proliferating cells is reduced by at least 5%; more preferably, by
at least 10%; more preferably, by at least 20%; more preferably, by
at least 30%; more preferably, by at least 40%; more preferably, by
at least 50%; even more preferably, by at least 50%; and most
preferably, by at least 75%. The proportion of proliferating cells
may be measured by any reproducible means of measurement. In a
preferred aspect, the proportion of proliferating cells is
measured, for example, by quantifying the number of dividing cells
relative to the number of nondividing cells in a tissue sample. In
another preferred aspect, the proportion of proliferating cells is
equivalent to the mitotic index.
[0170] In another aspect, treating or preventing a cell
proliferative disorder results in a decrease in size of an area or
zone of cellular proliferation. Preferably, after treatment, size
of an area or zone of cellular proliferation is reduced by at least
5% relative to its size prior to treatment; more preferably,
reduced by at least 10%; more preferably, reduced by at least 20%;
more preferably, reduced by at least 30%; more preferably, reduced
by at least 40%; more preferably, reduced by at least 50%; even
more preferably, reduced by at least 50%; and most preferably,
reduced by at least 75%. Size of an area or zone of cellular
proliferation may be measured by any reproducible means of
measurement. In a preferred aspect, size of an area or zone of
cellular proliferation may be measured as a diameter or width of an
area or zone of cellular proliferation.
[0171] In another aspect, treating or preventing a cell
proliferative disorder results in a decrease in the number or
proportion of cells having an abnormal appearance or morphology.
Preferably, after treatment, the number of cells having an abnormal
morphology is reduced by at least 5% relative to its size prior to
treatment; more preferably, reduced by at least 10%; more
preferably, reduced by at least 20%; more preferably, reduced by at
least 30%; more preferably, reduced by at least 40%; more
preferably, reduced by at least 50%; even more preferably, reduced
by at least 50%; and most preferably, reduced by at least 75%. An
abnormal cellular appearance or morphology may be measured by any
reproducible means of measurement. In one aspect, an abnormal
cellular morphology is measured by microscopy, e.g., using an
inverted tissue culture microscope. In one aspect, an abnormal
cellular morphology takes the form of nuclear pleiomorphism.
[0172] As used herein, the term "selectively" means tending to
occur at a higher frequency in one population than in another
population. In one aspect, the compared populations are cell
populations. In a preferred aspect, a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof, acts selectively on a
cancer or precancerous cell but not on a normal cell. In another
preferred aspect, a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or
derivative thereof, acts selectively to modulate one molecular
target (e.g., c-Met) but does not significantly modulate another
molecular target (e.g., Protein Kinase C). In another preferred
aspect, the invention provides a method for selectively inhibiting
the activity of an enzyme, such as a kinase. Preferably, an event
occurs selectively in population A relative to population B if it
occurs greater than two times more frequently in population A as
compared to population B. More preferably, an event occurs
selectively if it occurs greater than five times more frequently in
population A. More preferably, an event occurs selectively if it
occurs greater than ten times more frequently in population A; more
preferably, greater than fifty times; even more preferably, greater
than 100 times; and most preferably, greater than 1000 times more
frequently in population A as compared to population B. For
example, cell death would be said to occur selectively in cancer
cells if it occurred greater than twice as frequently in cancer
cells as compared to normal cells.
[0173] In a preferred aspect, a compound of the present invention
or a pharmaceutically acceptable salt, prodrug, metabolite, analog
or derivative thereof, modulates the activity of a molecular target
(e.g., c-Met). In one aspect, modulating refers to stimulating or
inhibiting an activity of a molecular target. Preferably, a
compound of the present invention modulates the activity of a
molecular target if it stimulates or inhibits the activity of the
molecular target by at least 2-fold relative to the activity of the
molecular target under the same conditions but lacking only the
presence of said compound. More preferably, a compound of the
present invention modulates the activity of a molecular target if
it stimulates or inhibits the activity of the molecular target by
at least 5-fold, at least 10-fold, at least 20-fold, at least
50-fold, at least 100-fold relative to the activity of the
molecular target under the same conditions but lacking only the
presence of said compound. The activity of a molecular target may
be measured by any reproducible means. The activity of a molecular
target may be measured in vitro or in vivo. For example, the
activity of a molecular target may be measured in vitro by an
enzymatic activity assay or a DNA binding assay, or the activity of
a molecular target may be measured in vivo by assaying for
expression of a reporter gene.
[0174] In one aspect, a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or
derivative thereof, does not significantly modulate the activity of
a molecular target if the addition of the compound does not
stimulate or inhibit the activity of the molecular target by
greater than 10% relative to the activity of the molecular target
under the same conditions but lacking only the presence of said
compound. In a preferred aspect, a compound of the present
invention does not significantly modulate the activity of Protein
Kinase C.
[0175] As used herein, the term "isozyme selective" means
preferential inhibition or stimulation of a first isoform of an
enzyme in comparison to a second isoform of an enzyme (e.g.,
preferential inhibition or stimulation of a kinase isozyme alpha in
comparison to a kinase isozyme beta). Preferably, a compound of the
present invention demonstrates a minimum of a four fold
differential, preferably a ten fold differential, more preferably a
fifty fold differential, in the dosage required to achieve a
biological effect. Preferably, a compound of the present invention
demonstrates this differential across the range of inhibition, and
the differential is exemplified at the IC so, i.e., a 50%
inhibition, for a molecular target of interest.
[0176] In a preferred embodiment, administering a compound of the
present invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof, to a cell or a subject in
need thereof results in modulation (i.e., stimulation or
inhibition) of an activity of c-Met. As used herein, an activity of
c-Met refers to any biological function or activity that is carried
out by c-Met. For example, a function of c-Met includes
phosphorylation of downstream target proteins. Other functions of
c-Met include autophosphorylation, binding of adaptor proteins such
as Gab-1, Grb-2, She, SHP2 and c-Cbl, and activation of signal
transducers such as Ras, Src, PI3K, PLC-.gamma., STATs, ERK1 and 2
and FAK.
[0177] In a preferred embodiment, administering a compound of the
present invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof, to a cell or a subject in
need thereof results in modulation (i.e., stimulation or
inhibition) of an activity of ERK 1 or ERK 2, or both. As used
herein, an activity of ERK 1 or ERK 2 refers to any biological
function or activity that is carried out by ERK 1 or ERK 2. For
example, a function of ERK 1 or ERK 2 includes phosphorylation of
downstream target proteins.
[0178] In one aspect, activating refers to placing a composition of
matter (e.g., protein or nucleic acid) in a state suitable for
carrying out a desired biological function. In one aspect, a
composition of matter capable of being activated also has an
unactivated state. In one aspect, an activated composition of
matter may have an inhibitory or stimulatory biological function,
or both.
[0179] In one aspect, elevation refers to an increase in a desired
biological activity of a composition of matter (e.g., a protein or
a nucleic acid). In one aspect, elevation may occur through an
increase in concentration of a composition of matter.
[0180] As used herein, "a cell cycle checkpoint pathway" refers to
a biochemical pathway that is involved in modulation of a cell
cycle checkpoint. A cell cycle checkpoint pathway may have
stimulatory or inhibitory effects, or both, on one or more
functions comprising a cell cycle checkpoint. A cell cycle
checkpoint pathway is comprised of at least two compositions of
matter, preferably proteins, both of which contribute to modulation
of a cell cycle checkpoint. A cell cycle checkpoint pathway may be
activated through an activation of one or more members of the cell
cycle checkpoint pathway. Preferably, a cell cycle checkpoint
pathway is a biochemical signaling pathway.
[0181] As used herein, "cell cycle checkpoint regulator" refers to
a composition of matter that can function, at least in part, in
modulation of a cell cycle checkpoint. A cell cycle checkpoint
regulator may have stimulatory or inhibitory effects, or both, on
one or more functions comprising a cell cycle checkpoint. In one
aspect, a cell cycle checkpoint regulator is a protein. In another
aspect, a cell cycle checkpoint regulator is not a protein.
[0182] In one aspect, treating cancer or a cell proliferative
disorder results in cell death, and preferably, cell death results
in a decrease of at least 10% in number of cells in a population.
More preferably, cell death means a decrease of at least 20%; more
preferably, a decrease of at least 30%; more preferably, a decrease
of at least 40%; more preferably, a decrease of at least 50%; most
preferably, a decrease of at least 75%. Number of cells in a
population may be measured by any reproducible means. In one
aspect, number of cells in a population is measured by fluorescence
activated cell sorting (FACS). In another aspect, number of cells
in a population is measured by immunofluorescence microscopy. In
another aspect, number of cells in a population is measured by
light microscopy. In another aspect, methods of measuring cell
death are as shown in Li et al., (2003) Proc Natl Acad Sci USA.
100(5): 2674-8. In an aspect, cell death occurs by apoptosis.
[0183] In a preferred aspect, an effective amount of a compound of
the present invention, or a pharmaceutically acceptable salt,
prodrug, metabolite, analog or derivative thereof is not
significantly cytotoxic to normal cells. A therapeutically
effective amount of a compound is not significantly cytotoxic to
normal cells if administration of the compound in a therapeutically
effective amount does not induce cell death in greater than 10% of
normal cells. A therapeutically effective amount of a compound does
not significantly affect the viability of normal cells if
administration of the compound in a therapeutically effective
amount does not induce cell death in greater than 10% of normal
cells. In an aspect, cell death occurs by apoptosis.
[0184] In one aspect, contacting a cell with a compound of the
present invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof, induces or activates cell
death selectively in cancer cells. Preferably, administering to a
subject in need thereof a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or
derivative thereof, induces or activates cell death selectively in
cancer cells. In another aspect, contacting a cell with a compound
of the present invention, or a pharmaceutically acceptable salt,
prodrug, metabolite, analog or derivative thereof, induces cell
death selectively in one or more cells affected by a cell
proliferative disorder. Preferably, administering to a subject in
need thereof a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or
derivative thereof, induces cell death selectively in one or more
cells affected by a cell proliferative disorder. In a preferred
aspect, the present invention relates to a method of treating or
preventing cancer by administering a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof to a subject in need
thereof, where administration of the compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof results in one or more of
the following: accumulation of cells in G1 and/or S phase of the
cell cycle, cytotoxicity via cell death in cancer cells without a
significant amount of cell death in normal cells, antitumor
activity in animals with a therapeutic index of at least 2, and
activation of a cell cycle checkpoint. As used herein, "therapeutic
index" is the maximum tolerated dose divided by the efficacious
dose.
[0185] One skilled in the art may refer to general reference texts
for detailed descriptions of known techniques discussed herein or
equivalent techniques. These texts include Ausubel et al., Current
Protocols in Molecular Biology, John Wiley and Sons, Inc. (2005);
Sambrook et al., Molecular Cloning, A Laboratory Manual (3d ed.),
Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (2000); Coligan
et al., Current Protocols in Immunology, John Wiley & Sons,
N.Y.; Enna et al., Current Protocols in Pharmacology, John Wiley
& Sons, N.Y.; Fingl et al., The Pharmacological Basis of
Therapeutics (1975), Remington's Pharmaceutical Sciences, Mack
Publishing Co., Easton, Pa., 18th edition (1990). These texts can,
of course, also be referred to in making or using an aspect of the
invention.
[0186] In additional aspects, a compound of the present invention,
or a pharmaceutically acceptable salt, prodrug, metabolite, analog
or derivative thereof, may be administered in combination with a
second chemotherapeutic agent. The second chemotherapeutic agent
can be a taxane, an aromatase inhibitor, an anthracycline, a
microtubule targeting drug, a topoisomerase poison drug, a targeted
monoclonal or polyconal antibody, an inhibitor of a molecular
target or enzyme (e.g., a kinase inhibitor), or a cytidine analogue
drug. In preferred aspects, the chemotherapeutic agent can be, but
not restricted to, tamoxifen, raloxifene, anastrozole, exemestane,
letrozole, HERCEPTIN.RTM. (trastuzumab), GLEEVEC.RTM. (imatinib),
TAXOL.RTM. (paclitaxel), cyclophosphamide, lovastatin, minosine,
araC, 5-fluorouracil (5-FU), methotrexate (MTX), TAXOTERE.RTM.
(docetaxel), ZOLADEX.RTM. (goserelin), vincristine, vinblastine,
nocodazole, teniposide, etoposale, GEMZAR.RTM. (gemcitabine),
epothilone, navelbine, camptothecin, daunorubicin, dactinomycin,
mitoxantrone, amsacrine, doxorubicin (adriamycin), epirubicin or
idarubicin or agents listed in
www.cancer.org/docroot/cdg/cdg_.asp.
[0187] In another aspect, the second chemotherapeutic agent can be
a cytokine such as G-CSF (granulocyte colony stimulating factor).
In another aspect, a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or
derivative thereof, may be administered in combination with
radiation therapy. In yet another aspect, a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof, may be administered in
combination with standard chemotherapy combinations such as, but
not restricted to, CMF (cyclophosphamide, methotrexate and
5-fluorouracil), CAF (cyclophosphamide, adriamycin and
5-fluorouracil), AC (adriamycin and cyclophosphamide), FEC
(5-fluorouracil, epirubicin, and cyclophosphamide), ACT or ATC
(adriamycin, cyclophosphamide, and paclitaxel), or CMFP
(cyclophosphamide, methotrexate, 5-fluorouracil and
prednisone).
[0188] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, analog or derivative thereof,
can be incorporated into pharmaceutical compositions suitable for
administration. Such compositions typically comprise the compound
(i.e. including the active compound), and a pharmaceutically
acceptable excipient or carrier. As used herein, "pharmaceutically
acceptable excipient" or "pharmaceutically acceptable carrier" is
intended to include any and all solvents, dispersion media,
coatings, antibacterial and antifungal agents, isotonic and
absorption delaying agents, and the like, compatible with
pharmaceutical administration. Suitable carriers are described in
the most recent edition of Remington's Pharmaceutical Sciences, a
standard reference text in the field. Preferred examples of such
carriers or diluents include, but are not limited to, water,
saline, ringer's solutions, dextrose solution, and 5% human serum
albumin. Pharmaceutically acceptable carriers include solid
carriers such as lactose, terra alba, sucrose, talc, gelatin, agar,
pectin, acacia, magnesium stearate, stearic acid and the like.
Exemplary liquid carriers include syrup, peanut oil, olive oil,
water and the like. Similarly, the carrier or diluent may include
time-delay material known in the art, such as glyceryl monostearate
or glyceryl distearate, alone or with a wax, ethylcellulose,
hydroxypropylmethylcellulose, methylmethacrylate or the like. Other
fillers, excipients, flavorants, and other additives such as are
known in the art may also be included in a pharmaceutical
composition according to this invention. Liposomes and non-aqueous
vehicles such as fixed oils may also be used. The use of such media
and agents for pharmaceutically active substances is well known in
the art. Except insofar as any conventional media or agent is
incompatible with the active compound, use thereof in the
compositions is contemplated. Supplementary active compounds can
also be incorporated into the compositions.
[0189] In one aspect, a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or
derivative thereof, is administered in a suitable dosage form
prepared by combining a therapeutically effective amount (e.g. an
efficacious level sufficient to achieve the desired therapeutic
effect through inhibition of tumor growth, killing of tumor cells,
treatment or prevention of cell proliferative disorders, etc.) of a
compound of the present invention, or a pharmaceutically acceptable
salt, prodrug, metabolite, analog or derivative thereof, (as an
active ingredient) with standard pharmaceutical carriers or
diluents according to conventional procedures (i.e., by producing a
pharmaceutical composition of the invention). These procedures may
involve mixing, granulating, and compressing or dissolving the
ingredients as appropriate to attain the desired preparation.
4. The Pharmaceutical Compositions and Formulations
[0190] A pharmaceutical composition of the invention is formulated
to be compatible with its intended route of administration.
Examples of routes of administration include parenteral, e.g.,
intravenous, intradermal, subcutaneous, oral (e.g., inhalation),
transdermal (topical), and transmucosal administration. Solutions
or suspensions used for parenteral, intradermal, or subcutaneous
application can include the following components: a sterile diluent
such as water for injection, saline solution, fixed oils,
polyethylene glycols, glycerine, propylene glycol or other
synthetic solvents; antibacterial agents such as benzyl alcohol or
methyl parabens; antioxidants such as ascorbic acid or sodium
bisulfite; chelating agents such as ethylenediaminetetraacetic
acid; buffers such as acetates, citrates or phosphates, and agents
for the adjustment of tonicity such as sodium chloride or dextrose.
The pH can be adjusted with acids or bases, such as hydrochloric
acid or sodium hydroxide. The parenteral preparation can be
enclosed in ampoules, disposable syringes or multiple dose vials
made of glass or plastic.
[0191] A compound or pharmaceutical composition of the invention
can be administered to a subject in many of the well-known methods
currently used for chemotherapeutic treatment. For example, for
treatment of cancers, a compound of the invention may be injected
directly into tumors, injected into the blood stream or body
cavities or taken orally or applied through the skin with patches.
The dose chosen should be sufficient to constitute effective
treatment but not so high as to cause unacceptable side effects.
The state of the disease condition (e.g., cancer, precancer, and
the like) and the health of the patient should preferably be
closely monitored during and for a reasonable period after
treatment.
[0192] The term "therapeutically effective amount," as used herein,
refers to an amount of a pharmaceutical agent to treat, ameliorate,
or prevent an identified disease or condition, or to exhibit a
detectable therapeutic or inhibitory effect. The effect can be
detected by any assay method known in the art. The precise
effective amount for a subject will depend upon the subject's body
weight, size, and health; the nature and extent of the condition;
and the therapeutic or combination of therapeutics selected for
administration. Therapeutically effective amounts for a given
situation can be determined by routine experimentation that is
within the skill and judgment of the clinician. In a preferred
aspect, the disease or condition to be treated is cancer. In
another aspect, the disease or condition to be treated is a cell
proliferative disorder.
[0193] For any compound, the therapeutically effective amount can
be estimated initially either in cell culture assays, e.g., of
neoplastic cells, or in animal models, usually rats, mice, rabbits,
dogs, or pigs. The animal model may also be used to determine the
appropriate concentration range and route of administration. Such
information can then be used to determine useful doses and routes
for administration in humans. Therapeutic/prophylactic efficacy and
toxicity may be determined by standard pharmaceutical procedures in
cell cultures or experimental animals, e.g., ED.sub.50 (the close
therapeutically effective in 50% of the population) and LD.sub.50
(the dose lethal to 50% of the population). The close ratio between
toxic and therapeutic effects is the therapeutic index, and it can
be expressed as the ratio, LD.sub.50/ED.sub.50. Pharmaceutical
compositions that exhibit large therapeutic indices are preferred.
The dosage may vary within this range depending upon the dosage
form employed, sensitivity of the patient, and the route of
administration.
[0194] Dosage and administration are adjusted to provide sufficient
levels of the active agent(s) or to maintain the desired effect.
Factors which may be taken into account include the severity of the
disease state, general health of the subject, age, weight, and
gender of the subject, diet, time and frequency of administration,
drug combination(s), reaction sensitivities, and tolerance/response
to therapy. Long-acting pharmaceutical compositions may be
administered every 3 to 4 days, every week, or once every two weeks
depending on half-life and clearance rate of the particular
formulation.
[0195] The pharmaceutical compositions containing active compounds
of the present invention may be manufactured in a manner that is
generally known, e.g., by means of conventional mixing, dissolving,
granulating, dragee-making, levigating, emulsifying, encapsulating,
entrapping, or lyophilizing processes. Pharmaceutical compositions
may be formulated in a conventional manner using one or more
pharmaceutically acceptable carriers comprising excipients and/or
auxiliaries that facilitate processing of the active compounds into
preparations that can be used pharmaceutically. Of course, the
appropriate formulation is dependent upon the route of
administration chosen.
[0196] Pharmaceutical compositions suitable for injectable use
include sterile aqueous solutions (where water soluble) or
dispersions and sterile powders for the extemporaneous preparation
of sterile injectable solutions or dispersion. For intravenous
administration, suitable carriers include physiological saline,
bacteriostatic water, Cremophor EL.TM. (BASF, Parsippany, N.J.) or
phosphate buffered saline (PBS). In all cases, the composition must
be sterile and should be fluid to the extent that easy
syringeability exists. It must be stable under the conditions of
manufacture and storage and must be preserved against the
contaminating action of microorganisms such as bacteria and fungi.
The carrier can be a solvent or dispersion medium containing, for
example, water, ethanol, polyol (for example, glycerol, propylene
glycol, and liquid polyethylene glycol, and the like), and suitable
mixtures thereof. The proper fluidity can be maintained, for
example, by the use of a coating such as lecithin, by the
maintenance of the required particle size in the case of dispersion
and by the use of surfactants. Prevent ion of the action of
microorganisms can be achieved by various antibacterial and
antifungal agents, for example, parabens, chlorobutanol, phenol,
ascorbic acid, thimerosal, and the like. In many cases, it will be
preferable to include isotonic agents, for example, sugars,
polyalcohols such as manitol, sorbitol, sodium chloride in the
composition. Prolonged absorption of the injectable compositions
can be brought about by including in the composition an agent which
delays absorption, for example, aluminum monostearate and
gelatin.
[0197] Sterile injectable solutions can be prepared by
incorporating the active compound in the required amount in an
appropriate solvent with one or a combination of ingredients
enumerated above, as required, followed by filtered sterilization.
Generally, dispersions are prepared by incorporating the active
compound into a sterile vehicle that contains a basic dispersion
medium and the required other ingredients from those enumerated
above. In the case of sterile powders for the preparation of
sterile injectable solutions, methods of preparation are vacuum
drying and freeze-drying that yields a powder of the active
ingredient plus any additional desired ingredient from a previously
sterile-filtered solution thereof.
[0198] Oral compositions generally include an inert diluent or an
edible pharmaceutically acceptable carrier. They can be enclosed in
gelatin capsules or compressed into tablets. For the purpose of
oral therapeutic administration, the active compound can be
incorporated with excipients and used in the form of tablets,
troches, or capsules. Oral compositions can also be prepared using
a fluid carrier for use as a mouthwash, wherein the compound in the
fluid carrier is applied orally and swished and expectorated or
swallowed. Pharmaceutically compatible binding agents, and/or
adjuvant materials can be included as part of the composition. The
tablets, pills, capsules, troches and the like can contain any of
the following ingredients, or compounds of a similar nature: a
binder such as microcrystalline cellulose, gum tragacanth or
gelatin; an excipient such as starch or lactose, a disintegrating
agent such as alginic acid, Primogel, or corn starch; a lubricant
such as magnesium stearate or Sterotes; a glidant such as colloidal
silicon dioxide; a sweetening agent such as sucrose or saccharin;
or a flavoring agent such as peppermint, methyl salicylate, or
orange flavoring.
[0199] For administration by inhalation, the compounds are
delivered in the form of an aerosol spray from pressured container
or dispenser, which contains a suitable propellant, e.g., a gas
such as carbon dioxide, or a nebulizer.
[0200] Systemic administration can also be by transmucosal or
transdermal means. For transmucosal or transdermal administration,
penetrants appropriate to the barrier to be permeated are used in
the formulation. Such penetrants are generally known in the art,
and include, for example, for transmucosal administration,
detergents, bile salts, and fusidic acid derivatives. Transmucosal
administration can be accomplished through the use of nasal sprays
or suppositories. For transdermal administration, the active
compounds are formulated into ointments, salves, gels, or creams as
generally known in the art.
[0201] In one aspect, the active compounds are prepared with
pharmaceutically acceptable carriers that will protect the compound
against rapid elimination from the body, such as a controlled
release formulation, including implants and microencapsulated
delivery systems. Biodegradable, biocompatible polymers can be
used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic
acid, collagen, polyorthoesters, and polylactic acid. Methods for
preparation of such formulations will be apparent to those skilled
in the art. The materials can also be obtained commercially from
Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal
suspensions (including liposomes targeted to infected cells with
monoclonal antibodies to viral antigens) can also be used as
pharmaceutically acceptable carriers. These can be prepared
according to methods known to those skilled in the art, for
example, as described in U.S. Pat. No. 4,522,811.
[0202] It is especially advantageous to formulate oral or
parenteral compositions in dosage unit form for ease of
administration and uniformity of dosage. Dosage unit form as used
herein refers to physically discrete units suited as unitary
dosages for the subject to be treated; each unit containing a
predetermined quantity of active compound calculated to produce the
desired therapeutic effect in association with the required
pharmaceutical carrier. The specification for the dosage unit forms
of the invention are dictated by and directly dependent on the
unique characteristics of the active compound and the particular
therapeutic effect to be achieved.
[0203] In therapeutic applications, the dosages of the
pharmaceutical compositions used in accordance with the invention
vary depending on the agent, the age, weight, and clinical
condition of the recipient patient, and the experience and judgment
of the clinician or practitioner administering the therapy, among
other factors affecting the selected dosage. Generally, the dose
should be sufficient to result in slowing, and preferably
regressing, the growth of the tumors and also preferably causing
complete regression of the cancer. Dosages can range from about
0.01 mg/kg per day to about 3000 mg/kg per day. In preferred
aspects, dosages can range from about 1 mg/kg per day to about 1000
mg/kg per day. In an aspect, the lose will be in the range of about
0.1 mg/day to about 50 g/day; about 0.1 mg/day to about 25 g/day;
about 0.1 mg/day to about 10 g/day; about 0.1 mg to about 3 g/day;
or about 0.1 mg to about 1 g/day, in single, divided, or continuous
doses (which dose may be adjusted for the patient's weight in kg,
body surface area in min, and age in years). An effective amount of
a pharmaceutical agent is that which provides an objectively
identifiable improvement as noted by the clinician or other
qualified observer. For example, regression of a tumor in a patient
may be measured with reference to the diameter of a tumor. Decrease
in the diameter of a tumor indicates regression. Regression is also
indicated by failure of tumors to reoccur after treatment has
stopped. As used herein, the term "dosage effective manner" refers
to amount of an active compound to produce the desired biological
effect in a subject or cell.
[0204] The pharmaceutical compositions can be included in a
container, pack, or dispenser together with instructions for
administration.
[0205] All patents, patent applications and references cited herein
are incorporated by reference herein in their entirety.
EXAMPLES
[0206] Examples are provided below to further illustrate different
features of the present invention. The examples also illustrate
useful methodology for practicing the invention. These examples do
not limit the claimed invention.
Example 1
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione
Step 1
##STR00008##
[0208] To a solution of 1,2,3,4-tetrahydroquinoline (100 ml) in
anhydrous tetrahydrofuran (300 ml), bromoethylpyrruvate (53 ml) was
added dropwise over 30 minutes. The mixture was stirred for 24
hours at room temperature. The reaction mixture was filtered and
the solid washed with tetrahydrofuran (100 ml). The filtrate was
evaporated to dryness to give
3-(3,4-dihydro-2H-quinolin-1-yl)-2-oxopropionic acid ethyl ester as
a brown oil 117 g.
Step 2
##STR00009##
[0210] Anhydrous magnesium chloride (29.4 g, 0.31 mol) was
suspended in 2-methoxyethanol (400 ml), and the mixture was stirred
for 15 minutes at 125.degree. C. A solution of
3-(3,4-dihydro-2H-quinolin-1-yl)-2-oxopropionic acid ethyl ester
(76.57 g 0.31 mol) in 2-methoxyethanol (100 ml) was then added and
the mixture stirred at 125.degree. C. for 60 minutes. The mixture
was stirred for a further 5 hours at reflux, cooled and evaporated
to dryness. The residue was then acidified with 2 M hydrochloric
acid (500 ml) and extracted with dichloromethane (3.times.500 ml).
The combined organic layers were then washed with 5% sodium
bicarbonate solution and dried over anhydrous magnesium sulfate
before being evaporated to dryness. The residue was then purified
on a silica gel chromatography column, eluting with ethyl
acetate/hexanes (1:4) to provide 5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinoline-1-carboxylic acid ethyl ester (31.0 g, 47%). .sup.1H NMR
(CDCl.sub.3) 400 MHz .delta.: 7.9 (d, 1H, J=8 Hz), 7.79 (s, 1H),
7.17 (m, 1H), 6.99 (d, 1H, J=7.2 Hz), 4.37 (m, 2H), 4.18 (t, 2H,
J=5.6 Hz), 3.0 (t, 2H, J=6 Hz), 2.24 (t, 2H, J=6 Hz), 1.42 (t, 3H,
J=7.2 Hz).
Step 3
##STR00010##
[0212] To a solution of 5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinoline-1-carboxylic acid ethyl ester (31 g, 0.14 mol) in ethanol
(200 ml) and water (200 ml) was added sodium hydroxide (30.8 g,
0.77 mol). The mixture was heated to reflux for 2 hours before
being cooled to room temperature and diluted with water (2.64 L).
The mixture was then washed with dichloromethane (2.times.300 ml)
and the aqueous layer was acidified with concentrated hydrochloric
acid to pH 1.0. The precipitate formed was collected by filtration,
washed with water and dried to yield 5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinoline-1-carboxylic acid as a dark yellow solid (23
g, 85%). .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.95 (brs,
1H), 7.96 (s, 1H), 7.69 (d, 1H, J=8.4 Hz), 7.06 (t, 1H, J=6.8 Hz),
6.92 (d, 1H, J=6.8 Hz), 4.19 (t, 2H, J=6 Hz), 2.91 (t, 2H, J=6 Hz),
2.11 (t, 2H, J=5.6 Hz).
Step 4
##STR00011##
[0214] 5,6-dihydro-4H-pyrrolo [3,2,1-ij] quinoline-1-carboxylic
acid (37.5 g, 0.186 mol), copper chromite (13.5 g, 43 mmol) and
quinoline (180 ml) were heated with stirring to 185.degree. C. for
2 hours. The mixture was cooled, diluted with dichloromethane (1 L)
and filtered over hyflo. The filtrate was washed with 2 M
hydrochloric acid (2.times.600 ml) and twice with 2 M sodium
hydroxide (150 ml) before being evaporated to dryness. The residue
was purified by silica gel chromatography, eluting with a ethyl
acetate/hexanes (1:6) to afford 5, 6-dihydro-4H-pyrrolo [3,2,1-ij]
quinoline (21 g, 72%) as a pale yellow solid. .sup.1H NMR
(CDCl.sub.3) 400 MHz .delta.: 7.44 (dd, 1H, J=0.8 and 7.6 Hz), 7.07
(d, 1H, J=3.2 Hz), 7.01 (t, 1H, J=7.2 Hz), 6.9 (dd, 1H, J=0.8 and
6.8 Hz), 6.43 (d, 1H, J=3.2 Hz), 4.16 (t, 2H, J=6 Hz), 2.99 (t, 2H,
3=6.4 Hz), 2.24 (m, 2H).
##STR00012##
[0215] To a solution of 5, 6-dihydro-4H-pyrroloquinoline (4.0 g,
25.3 mmol), in anhydrous ether (300 ml) at 0.degree. C., was added
oxalyl chloride (2.22 ml, 25.3 mmol). The mixture was stirred for
30-45 minutes at 0.degree. C. before being cooled to -78.degree. C.
Sodium methoxide in methanol (0.5M) (60 ml) was then added slowly
and the mixture allowed to warm to room temperature. The mixture
was then diluted with ethyl acetate (200 ml), washed with water
(100 ml) followed by a wash with saturated aqueous sodium chloride
(50 ml). The organic layer was dried over anhydrous sodium sulfate
and evaporated to dryness. The residue was dissolved in ethyl
acetate (100 ml) filtered through a 2 inch plug of coarse silica
gel and evaporated to give 5, 6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl) oxoacetic acid methyl ester as a yellow solid (5.3
g, 85%). .sup.1H NMR (CDCl.sub.3) 400 MHz .delta.: 8.3 (s, 1H),
8.14 (d, 1H), 7.22 (t, 1H), 7.04 (d, 1H), 4.2 (t, 2H), 3.95 (s,
3H), 3.0 (t, 2H), 2.3 (t, 2H).
Step 6
##STR00013##
[0217] To a solution of 5, 6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl) oxoacetic acid methyl ester (1.0 g, 4.12 mmol) and
indole-3-acetamide (0.8 g, 4.5 mmol) in anhydrous tetrahydrofuran
at 0.degree. C. was added a solution of potassium t-butoxide (1M in
tetrahydrofuran) (12.4 ml, 12.4 mmol) dropwise over 30 minutes. The
mixture was stirred at 0.degree. C. for 2 hours. Concentrated
hydrochloric acid (10 ml) was then added and the mixture stirred
for 1 hour at room temperature. The mixture was then diluted with
ethyl acetate (200 ml), washed twice with water (50 ml), and
saturated aqueous sodium chloride solution (50 ml) and the organic
layer dried over anhydrous sodium sulfate. The residue was purified
by silica gel chromatography, eluting with ethyl acetate/hexanes
(1:4) to afford 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione as a bright red
solid (1.2 g, 80%). .sup.1H NMR (CDCl.sub.3) 400 MHz .delta.: 8.5
(brs, 1H), 7.78 (s, 1H), 7.63 (d, 1H, J=2.8 Hz), 7.44 (s, 1H), 7.35
(d, 1H. J=8 Hz), 7.16 (d, 1H, J=8.4 Hz), 7.11 (t, 1H, J=7.6 Hz),
6.86 (t, 1H, J=7.6 Hz), 6.80 (d, 1H, J=7.2 Hz), 6.64 (t, 1H, J=8
Hz), 6.57 (d, 1H, J=8 Hz), 4.2 (t, 2H, J=6 Hz), 2.96 (t, 2H, J=6
Hz), 2.24 (m, 2H).
Example 2
Preparation of (.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione and
(.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
[0218] Preparation of the (.+-.)-cis compounds, (.+-.)-trans
compounds, or mixtures thereof were obtained using reducing
conditions as described in each of Procedures A through C.
Procedure A: Reduction of 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione with Zn/Hg
##STR00014##
[0220] The active zinc-mercury reducing agent for the reduction of
3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij] quinolin-1yl)-4(1H-indol-3-yl)
pyrrole-2, 5-dione was prepared from metallic zinc and HgCl.sub.2.
Zinc powder (2.5 g) and mercury (II) chloride (0.25 g) were
suspended in de-ionized water (3 ml) and stirred for 20 minutes. A
few drops of concentrated hydrochloric acid was then added and the
mixture stirred for few minutes. The solid was filtered off, washed
with de-ionized water (50 ml), ethanol (50 ml) and dried.
[0221] To a suspension of the Zn(Hg) reducing agent prepared as
above in dry ethanol (50 ml) was added 3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione (0.35
g, 95.4 .mu.mol). The mixture was heated to reflux for 30-60
minutes while dry hydrogen chloride gas was slowly passed through
the mixture. The mixture was then cooled, filtered, and evaporated
to dryness. A 5% potassium carbonate solution (150 ml) and ethyl
acetate (300 ml) were then added. The organic layer was dried over
anhydrous magnesium sulfate and evaporated to give .about.2:1
mixture of (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione and
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione (0.2 g).
Procedure B: Reduction of 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione with hydrogen in
the presence of palladium on carbon
##STR00015##
[0223] A suspension of 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione (16 g, 43.6
mmol) and 10% palladium on carbon (Pd/C, wet catalyst) (8 g) were
stirred under 1 atmosphere of hydrogen in methanol (600 ml) at room
temperature for 48 hours. The catalyst was then filtered through a
bed of Celite and the filtrate evaporated to dryness. The residue
was re-dissolved in methanol and the product precipitated by the
addition of cold water. The precipitate was filtered, washed with
water and dried under vacuum to yield
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione (9.2 g).
.sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.56 (s, 1H), 10.66
(s, 1H), 7.43 (d, 1H, J=7.6 Hz), 7.14 (d, 2H, J=8 Hz), 6.86-6.97
(m, 4H), 6.78 (t, 1H, J=7.2 Hz), 6.69 (d, 1H, J=6.8 Hz), 4.88 (dd,
2H, J=9.2 and 45.6 Hz), 3.88 (m, 2H), 2.76 (t, 2H, J=5.6 Hz), 1.94
(t, 2H, J=6 Hz).
Procedure C: Reduction of 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione by magnesium min
methanol
##STR00016##
[0225] Magnesium turnings (3.05 g, 0.125 mol) were added to a
solution of 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione (2.56 g, 6.97
mmol) in anhydrous methanol (100 ml) and heated to reflux under an
atmosphere of nitrogen for 40 minutes. After cooling to room
temperature the mixture was poured into ethyl acetate (300 ml),
washed with 1M hydrochloric acid (300 ml) and water (500 ml). The
organic layer was dried over anhydrous sodium sulfate and
evaporated to dryness. The residue was then purified by silica gel
chromatography using 40-50% ethyl acetate in hexanes to yield
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione as a pale
pink solid (2.3 g). .sup.1H-NMR (DMSO-d.sub.6) 400 MHz .delta.:
11.54 (s, 1H), 11.03 (s, 1H), 7.32-7.4 (m, 4H), 7.17 (d, 1H, J=7.2
Hz), 7.07 (t, 1H, J=7.6 Hz), 6.96 (t, 1H, J=7.6 Hz), 6.82-6.89 (m,
2H), 4.5 (dd, 2H, J=7.2 and 20 Hz), 4.07 (t, 2H, J=5.2 Hz), 2.87
(t, 2H, J=6 Hz), 2.08 (m, 2H).
Example 3
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione from
(.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
##STR00017##
[0227] A preparation of (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
(378 mg, 1.02 mmol) was heated to 50.degree. C. in tert-butanol (10
ml) and potassium t-butoxide (11 mg, 98 .mu.mol) for 16 hours. The
mixture was poured into ethyl acetate (100 ml) and washed with
water (100 ml). The organic layer was dried over anhydrous sodium
sulfate and evaporated to dryness to give
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione as a tan
powder (276 mg). .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.54
(s, 1H), 11.03 (s, 1H), 7.32-7.4 (m, 4H), 7.17 (d, 1H, J=7.2 Hz),
7.07 (t, 1H, J=7.6 Hz), 6.96 (t, 1H, J=7.6 Hz), 6.82-6.89 (m, 2H),
4.5 (dd, 2H, J=7.2 and 20 Hz), 4.07 (t, 2H, J=5.2 Hz), 2.87 (t, 2H,
J=6 Hz), 2.08 (m, 2H).
Example 4
Chromatographic Separation of 3(R),4(S)-3-(5,6-Dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
and 3(S),4(R)-3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
##STR00018##
[0229] A mixture of (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione (135 mg) in
methanol (10 ml) and acetonitrile (6 ml) was subjected to
preparative supercritical fluid chromatography, using a chiral AD
column 20 mm.times.250 mm, eluting with 35% methanol/CO2 at a flow
rate of 3.5 ml/minutes. To give a faster eluting peak at 4.55
minutes (60 mg) assigned 3(R),4(S)-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
and a slower eluting peak 6.05 minutes (56 mg), assigned
3(S),4(R)-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione. The
absolute stereochemical assignments were based solely upon the
relative retention time of related compounds and may be
reversed.
Example 5
Chromatographic Separation of 3(R),4(R)-3-(5,6-Dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
and 3(S),4(S)-3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
##STR00019##
[0231] A mixture of (.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
(200 mg) in acetonitrile (1 ml) was subjected to preparative
supercritical fluid chromatography using a CHIRALPAK.RTM. AD column
(Daicel, U.S.A.) 20 mm.times.250 mm, eluting with 35% methanol/CO2
at a flow rate of 3.5 ml/minutes. Chromatography yielded a faster
eluting peak of the trans isomer (82 mg) having a negative optical
rotation assigned (-)-3(R),4(R)-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
and a slower eluting peak of the trans isomer (86 mg) having a
positive optical rotation assigned
(+)-3(S),4(R)-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione. Absolute
stereochemical assignments were based solely upon relative
retention time of related compounds they may be reversed. All
optical rotation measurements were conducted in chloroform at
25.degree. C. at 589 nm.
[0232] Crystals of the chromatographically separated (+) or (-)
isomers of trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione may be
prepared from 2,2,2-trifluoroethanol using vapor stress techniques
and slow evaporation at 49.degree. C. Crystals of these isomers may
also be prepared from ethanol at room temperature by evaporation
employing seed crystals, such as those prepared by vapor stress
techniques.
Example 6
Preparation of 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(2-trifluoromethyl-phenyl)-pyrrole-2, 5-dione
[0233] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(2-trifluoromethyl-phenyl)-pyrrole-2, 5-dione was
prepared according to Example 1, steps 1-6, employing
2'-trifluoromethylphenyl acetamide in place of indole-3-acetamide
in step 6. .sup.1H NMR (CDCl.sub.3) 400 MHz .delta.: 8.16 (s, 1H),
7.83 (d, 2H, J=7.2 Hz), 7.58 (m, 2H), 7.37 (d, 1H, J=7.6 Hz), 7.33
(s, 1H), 6.85 (d, 1H, J=6.8 Hz), 6.66 (t 1H, J=7.2 Hz), 5.96 (d,
1H, J=8.8 Hz), 4.2 (t, 2H, J=5.6 Hz), 2.95 (t, 2H, J=6.4 Hz), 2.22
(m, 2H).
Example 7
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-thiophen-2-yl-pyrrole-2, 5-dione
[0234] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-thiophen-2-yl-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing 2-thienylacetamide in
place of indole-3-acetamide in step 6. .sup.1H NMR (CDCl.sub.3) 400
MHz .delta.: 7.87 (s, 1H), 7.49 (d, 1H, J=5.2 Hz), 7.37 (s, 1H),
7.3 (d, 1H, J=4 Hz), 7.02 (t, 1H, J=4 Hz), 6.89-6.98 (m, 2H), 6.53
(d, 1H, J=7.6 Hz), 4.92 (t, 2H, J=6 Hz), 3.04 (t, 2H, J=6 Hz), 2.31
(m, 2H).
Example 8
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(3-methoxy-phenyl)-pyrrole-2, 5-dione
[0235] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(3-methoxy-phenyl)-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing
3-methoxyphenylacetamide in place of indole-3-acetamide in step 6.
.sup.1H NMR (CDCl.sub.3) 400 MHz .delta.: 8.01 (s, 1H), 7.31 (s,
1H), 7.23 (t, 1H, J=7.6 Hz), 7.09 (m, 2H), 6.87-6.92 (m, 2H), 6.73
(t, 1H, J=7.6 Hz), 6.14 (d, 1H, J=8 Hz), 4.25 (t, 2H, J=5.2 Hz),
2.99 (t, 2H, J=5.6 Hz), 2.67 (m, 2H).
Example 9
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-pyridin-2-yl-pyrrole-2, 5-dione
[0236] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-pyridin-2-yl-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing pyridin-2-ylacetamide
in place of indole-3-acetamide in step 6. .sup.1H NMR (CDCl.sub.3)
400 MHz .delta.: 8.58 (d, 1H, J=4.4 Hz), 8.12 (s, 1H), 7.78 (dt,
1H, J=1.6 and 7.6 Hz), 7.68 (d, 1H, J=8 Hz), 7.31 (s, 1H), 7.25 (m,
1H), 6.87 (d, 1H, J=6 Hz), 6.68 (t, 1H, J=8 Hz), 5.91 (d, 1H, J=7.6
Hz), 4.24 (t, 2H, J=5.6 Hz), 2.97 (t, 2H, J=6 Hz), 2.25 (m,
2H).
Example 10
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(4-methoxy-phenyl)-pyrrole-2, 5-dione
[0237] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(4-methoxy-phenyl)-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing
4-methoxyphenylacetamide in place of indole-3-acetamide in step 6.
.sup.1H NMR (CDCl.sub.3) 400 MHz .delta.: 7.95 (s, 1H), 7.51 (m,
2H), 7.25 (s, 1H), 6.85-6.89 (m, 3H), 6.75 (t, 11-1, J=8 Hz), 6.24
(d, 1H, J=8 Hz), 4.26 (t, 2H, J=5.6 Hz), 3.82 (s, 3H), 2.99 (t, 2H,
J=6.4 Hz), 2.27 (m, 2H).
Example 11
Preparation of 3-Benzo[1,3]dioxol-5-yl-4-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-pyrrole-2, 5-dione
[0238] 3-Benzo[1,3]dioxol-5-yl-4-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-pyrrole-2, 5-dione was prepared according to Example
1, steps 1-6, employing 3,4-(methylenedioxy)phenylacetamide in
place of indole-3-acetamide in step 6. .sup.1H NMR (CDCl.sub.3) 400
MHz .delta.: 7.98 (s, 1H), 7.04-7.07 (m, 2H), 6.90 (d, 1H, J=7.2
Hz), 6.76-6.82 (m, 2H), 6.30 (d, 1H, J=8 Hz), 5.98 (s, 2H), 4.26
(t, 2H, J=5.6 Hz), 2.99 (t, 2H, J=6 Hz), 2.28 (m, 2H).
Example 12
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-phenyl-pyrrole-2, 5-dione
[0239] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-phenyl-pyrrole-2, 5-dione was prepared according
to Example 1, steps 1-6, employing phenylacetamide in place of
indole-3-acetamide in step 6. .sup.1H NMR (CDCl.sub.3) 400 MHz
.delta.: 8.01 (s, 1H), 7.52 (m, 2H), 7.35 (m, 3H), 7.27 (s, 1H),
6.87 (d, 1H, J=7.2 Hz), 6.7 (t, 1H, J=7.2 Hz), 6.08 (d, 1H, J=8
Hz), 4.26 (t, 2H, J=5.6 Hz), 2.99 (t, 2H, J=5.6 Hz), 2.27 (m,
2H).
Example 13
Preparation of 3-Benzo[b]thiophen-2-yl-4-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-pyrrole-2, 5-dione
[0240] 3-Benzo[b]thiophen-2-yl-4-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-pyrrole-2, 5-dione was prepared according to Example
1, steps 1-6, employing 2-benzothiophenylacetamide in place of
indole-3-acetamide in step 6. .sup.1H NMR (DMSO-d.sub.6) 400 MHz
.delta.: 11.11 (s, 1H), 8.14 (s, 1H), 8.01 (d, 1H, J=8 Hz), 7.84
(s, 1H), 7.45 (d, 1H, J=8 Hz), 7.3 (t, 1H, J=7.2 Hz), 7.15 (t, 1H,
J=7.6 Hz), 6.71 (d, 1H, J=6.8 Hz), 6.43 (t, 1H, J=7.6 Hz), 5.99 (d,
1H. J=8 Hz), 4.26 (t, 2H, J=5.2 Hz), 2.86 (t, 2H, J=5.6 Hz), 2.1
(m, 2H).
Example 14
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(3-phenoxy-phenyl)-pyrrole-2, 5-dione
[0241] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(3-phenoxy-phenyl)-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing
3-phenoxyphenylacetamide in place of indole-3-acetamide in step 6.
.sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.03 (s, 1H), 8.01 (s,
1H), 7.43 (t, 1H, J=7.6 Hz), 7.28 (d, 1H, J=7.6 Hz), 7.15 (t, 2H,
J=7.6 Hz), 7.03 (t, 2H, J=7.6 Hz), 6.92 (d, 1H, J=6.8 Hz), 6.8 (s,
1H), 6.76 (t, 1H, J=8 Hz), 6.60 (d, 2H, J=7.6 Hz), 6.08 (d, 1H, J=8
Hz), 4.27 (t, 2H, J=5.6 Hz), 2.97 (t, 2H, J=6 Hz), 2.16 (m,
2H).
Example 15
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(3-chloro-phenyl)-pyrrole-2, 5-dione
[0242] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(3-chloro-phenyl)-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing
3-chlorophenylacetamide in place of indole-3-acetamide in step 6.
.sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.11 (s, 1H), 8.13 (s,
1H), 7.47-7.43 (m, 2H), 7.36 (t, 11, J=7.6 Hz), 7.29 (d, 1H, J=7.6
Hz), 6.86 (d, 1H, J=6.8 Hz), 6.68 (t, 1H, J=7.6 Hz), 5.97 (d, 1H,
J=8 Hz), 4.31 (t, 2H, J=5.6 Hz), 2.93 (t, 2H, J=5.6 Hz), 2.16 (m,
2H).
Example 16
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2-chloro-phenyl)-pyrrole-2, 5-dione
[0243] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2-chloro-phenyl)-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing
2-chlorophenylacetamide in place of indole-3-acetamide in step 6.
.sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.1 (s, 1H), 8.17 (s,
1H), 7.55 (d, 1H, J=8 Hz), 7.45-7.49 (m, 1H), 7.36 (d, 2H, J=4.4
Hz), 6.81 (d, 1H, J=7.2 Hz), 6.58 (t, 1H, J=8 Hz), 5.92 (d, 1H,
J=8.4 Hz), 4.27 (m, 2H), 2.89 (t, 2H, J=6 Hz), 2.11 (m, 2H).
Example 17
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2,5-dimethoxy-phenyl)-pyrrole-2, 5-dione
[0244] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-yl)-4-(2,5-dimethoxy-phenyl)-pyrrole-2, 5-dione was
prepared according to Example 1, steps 1-6, employing
2,5-dimethoxyphenylacetamide in place of indole-3-acetamide in step
6. .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 10.93 (s, 1H), 8.06
(s, 1H), 6.97 (s, 2H), 6.81 (d, 1H, J=7.6 Hz), 6.77 (s, 1H), 6.6
(t, 1H, J=8 Hz), 5.92 (d, 1H, J=8 Hz), 4.26 (t, 2H, J=5.2 Hz), 3.63
(s, 3H), 3.3 (s, 3H), 2.9 (t, 2H, J=5.6 Hz), 2.11 (m, 2H).
Example 18
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2-chloro-4-fluoro-phenyl)-pyrrole-2, 5-dione
[0245] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2-chloro-4-fluoro-phenyl)-pyrrole-2, 5-dione was
prepared according to Example 1, steps 1-6, employing
2-chloro-4-fluorophenylacetamide in place of indole-3-acetamide in
step 6. .sup.1H NMR (DMSO-dd) 400 MHz .delta.: 11.11 (s, 1H), 8.16
(s, 1H), 7.57 (dd, 1H, J=2.8 and 9.2 Hz), 7.44 (dd, 1H, J=6.8 and
8.4 Hz), 7.28 (dt, 1H, J=2.4 and 8.4 Hz), 6.84 (d, 1H, J=7.2 Hz),
6.66 (t, 1H, J=8 Hz), 5.98 (d, 1H, J=8 Hz), 4.27 (m, 2H), 2.9 (t,
2H, J=5.6 Hz), 2.11 (m, 2H).
Example 19
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-naphthalene-1-yl-pyrrole-2, 5-dione
[0246] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-naphthalene-1-yl-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing 1-naphthylacetamide in
place of indole-3-acetamide in step 6. .sup.1H NMR (DMSO-d.sub.6)
400 MHz .delta.: 11.1 (s, 1H), 8.17 (s, 1H), 8.02 (d, 1H, J=8 Hz),
7.97 (d, 1H, J=8 Hz), 7.75 (d, 1H, J=8 Hz), 7.43-7.55 (m, 3H), 7.37
(t, 1H, J=8 Hz), 6.66 (d, 1H, J=6.8 Hz), 6.27 (t, 1H, J=8 Hz), 5.57
(d, 1H, J=8 Hz), 4.24 (t, 2H, J=5.2 Hz, 2.83 (t, 2H, J=5.6 Hz),
2.08 (m, 2H).
Example 20
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2,6-dichloro-phenyl)-pyrrole-2, 5-dione
[0247] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2,6-dichloro-phenyl)-pyrrole-2, 5-dione was
prepared according to Example 1, steps 1-6, employing
2,6-dichlorophenylacetamide in place of indole-3-acetamide in step
6. .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.23 (s, 1H), 8.27
(s, 1H), 7.53-7.62 (m, 3H), 6.85 (d, 1H, J=7.2 Hz), 6.64 (t, 1H,
J=8.4 Hz), 6.01 (d, 1H, J=8 Hz), 4.27 (t, 2H, J=5.6 Hz), 2.9 (t,
2H, J=5.6 Hz), 2.11 (m, 2H).
Example 21
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2-bromo-phenyl)-pyrrole-2, 5-dione
[0248] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2-bromo-phenyl)-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing 2-bromophenylacetamide
in place of indole-3-acetamide in step 6. .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 11.09 (s, 1H), 8.17 (s, 1H), 7.75
(m, 1H), 7.37 (m, 2H), 7.33 (m, 1H), 6.81 (d, 1H, J=7.2 Hz), 6.58
(t, 1H, J=8 Hz), 5.95 (d, 1H, J=8 Hz), 4.26 (t, 2H, J=5.6 Hz), 2.9
(t, 2H, J=5.6 Hz), 2.1 (m, 2H).
Example 22
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-indol-1-yl-pyrrole-2, 5-dione
[0249] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-indol-1-yl-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing N-indolyl-2-acetamide
in place of indole-3-acetamide in step 6. .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 11.21 (s, 1H), 8.18 (s, 1H), 7.58
(d, 1H, J=8 Hz), 7.52 (d, 1H, J=3.2 Hz), 7.01 (m, 2H), 6.91 (t, 1H,
J=6.8 Hz), 6.74 (d, 1H, J=2.8 Hz), 6.71 (d, 1H, J=7.2 Hz), 6.4 (t,
1H, J=8 Hz), 5.63 (d, 1H, J=8 Hz), 4.28 (t, 2H, J=4.8 Hz), 2.85 (t,
2H, J=5.6 Hz), 2.11 (m, 2H).
Example 23
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-pyridine-3-yl-pyrrole-2, 5-dione
[0250] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-pyridine-3-yl-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing pyridine-3-ylacetamide
in place of indole-3-acetamide in step 6. .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 11.14 (s, 1H), 8.53 (m, 2H), 8.12
(s, 1H), 7.78 (d, 1H, J=7.6 Hz), 7.41 (dd, 1H, J=4.8 and 8 Hz),
6.86 (d, 1H, J=7.2 Hz), 6.66 (t, 1H, 3=7.6 Hz), 5.97 (d, 1H, J=8.4
Hz), 4.3 (t, 2H, J=5.2 Hz), 2.93 (t, 2H, J=5.6 Hz), 2.16 (m,
2H).
Example 24
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(5-bromo-1H-indol-3-yl)-pyrrole-2, 5-dione
[0251] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(5-bromo-1H-indol-3-yl)-pyrrole-2, 5-dione was
prepared according to Example 1, steps 1-6, employing
5-bromo-1H-indoly-3-ylaccetamide in place of indole-3-acetamide in
step 6. .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.77 (s, 1H),
10.92 (s, 1H), 7.82 (s, 1H), 7.69 (d, 1H, J=2.4 Hz), 7.33 (d, 1H,
J=8.4 Hz), 7.10 (dd, 1H, J=2 and 8.4 Hz), 6.99 (d, 1H, J=1.6 Hz),
6.76 (d, 1H, J=7.2 Hz), 6.55 (t, 1H, J=8 Hz), 6.36 (d, 1H, J=8 Hz),
4.25 (t, 2H, J=5.6 Hz), 2.92 (t, 2H, J=5.6 Hz), 2.17 (m, 2H).
Example 25
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-pyridine-4-yl-pyrrole-2, 5-dione
[0252] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-pyridine-4-yl-pyrrole-2, 5-dione was prepared
according to Example 1, steps 1-6, employing pyridine-4-ylacetamide
in place of indole-3-acetamide in step 6. .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 11.17 (s, 1H), 8.53 (m, 2H), 8.54
(d, 2H, J=6 Hz), 8.17 (s, 1H), 7.32 (d, 2H, J=4.8 Hz), 6.88 (d, 1H,
J=7.2 Hz), 6.69 (t, 1H, J=7.6 Hz), 5.93 (d, 1H, J=8 Hz), 4.31 (t,
2H, J=6 Hz), 2.94 (t, 2H, 3=6 Hz), 2.16 (m, 2H).
Example 26
Preparation of 3-Biphenyl-4-yl-4-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-pyrrole-2, 5-dione
[0253] 3-Biphenyl-4-yl-4-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-pyrrole-2, 5-dione was prepared according to Example
1, steps 1-6, employing 4-phenylphenylacetamide in place of
indole-3-acetamide in step 6. .sup.1H NMR (acetone-d.sub.6) 400 MHz
.delta.: 8.08 (s, 1H), 7.6-7.73 (m, 7H), 7.48 (t, 2H, J=6.8 Hz),
7.39 (d, 1H, J=7.2 Hz), 6.84 (d, 1H, J=8 Hz), 6.65 (t, 1H, J=8.4
Hz), 6.23 (t, 1H, J=7.2 Hz), 5.97 (d, 1H, J=8.4 Hz), 4.38 (m, 2H),
2.98 (m, 2H), 2.28 (m, 2H).
Example 27
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(4-methanesulfonyl-phenyl)-pyrrole-2, 5-dione
[0254] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(4-methanesulfonyl-phenyl)-pyrrole-2, 5-dione was
prepared according to Example 1, steps 1-6, employing
4-methanesulfonylphenylacetamide in place of indole-3-acetamide in
step 6. .sup.1H NMR (CDCl.sub.3) 400 MHz .delta.: 8.09 (s, 1H), 7.9
(d, 2H, J=8.4 Hz), 7.71 (d, 2H, J=8.4 Hz), 7.64 (s, 1H), 6.91 (d,
1H, J=7.2 Hz), 6.73 (t, 1H, J=7.6 Hz), 5.95 (d, 1H, J=8.4 Hz), 4.29
(t, 2H, J=5.6 Hz), 3.06 (s, 3H), 3.0 (t, 2H, J=6 Hz), 2.29 (m,
2H).
Example 28
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2-trifluoromethyl-quinolin-4-yl-sulfanyl)-pyrrole-2,
5-dione
[0255] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-j]
quinolin-1-yl)-4-(2-trifluoromethyl-quinolin-4-yl-sulfanyl)-pyrrole-2,
5-dione was prepared according to Example 1, steps 1-6, employing
2-[[2-(trifluoromethyl)-4-quinolinyl]thio]acetamide in place of
indole-3-acetamide in step 6. .sup.1H NMR (CDCl.sub.3) 400 MHz
.delta.: 8.3 (d, 1H, J=7.6 Hz), 8.11 (d, 1H, J=8.4 Hz), 8.05 (s,
1H), 7.68-7.82 (m, 4H), 7.23 (s, 1H), 6.83 (m, 2H), 4.21 (t, 2H,
J=6 Hz), 2.92 (t, 2H, J=6 Hz), 2.21 (m, 2H).
Example 29
Preparation of 3-(4-Benzoyloxyphenyl)-4-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-pyrrole-2, 5-dione
[0256] 3-(4-Benzoyloxyphenyl)-4-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-pyrrole-2, 5-dione was prepared according to Example
1, steps 1-6, employing 4-benzyloxyphenylacetamide in place of
indole-3-acetamide in step 6. .sup.1H NMR (CDCl.sub.3) 400 MHz
.delta.: 7.95 (s, 1H), 7.5 (d, 2H, J=8.8 Hz), 7.33-7.43 (m, 6H),
6.93 (d, 2H, J=8.8 Hz), 6.88 (d, 1H, J=7.2 Hz), 6.73 (t, 1H, J=7.2
Hz), 6.23 (d, 1H, J=8.4 Hz), 5.08 (s, 2H), 4.25 (t, 2H, J=5.6 Hz),
2.99 (t, 2H, J=6 Hz), 2.27 (m, 2H).
Example 30
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-pyrrole-2,
5-dione
[0257] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1)-yl)-4-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-pyrrole-2,
5-dione was prepared according to Example 1, steps 1-6, employing
4-(2-morpholin-4-yl-ethoxy)-phenylacetamide in place of
indole-3-acetamide in step 6. .sup.1H NMR (CDCl.sub.3) 400 MHz
.delta.: 7.95 (s, 1H), 7.48 (m, 3H), 6.86 (m, 3H), 6.74 (t, 1H, J=8
Hz), 6.23 (d, 1H, J=8. Hz), 4.26 (t, 2H, J=5.2 Hz), 4.16 (t, 2H,
J=5.6 Hz), 3.77 (t, 4H, J=4.8 Hz), 2.99 (t, 2H, J=6 Hz), 2.87 (t,
2H, J=5.2 Hz), 2.65 (m, 4H), 2.28 (m, 2H).
Example 31
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-1-naphthyl-1H-indol-3-yl) pyrrole-2, 5-dione
[0258] A mixture of 1-naphthyl boronic acid (41 mg, 0.24 mmol),
3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-bromo-1H-indol-3-yl) pyrrole-2, 5-dione (88 mg,
0.2 mmol) (prepared as in Example 24), tetrakistriphenylphosphine
palladium (5 mol %) in toluene (4 ml), ethanol (4 ml), saturated
NaHCO.sub.3 (1 ml), and water (2 ml) was heated at 100.degree. C.
under nitrogen for 5 hours. After cooling to room temperature, the
mixture was extracted with ethyl acetate (3.times.15 ml) and
concentrated. The residue was purified by silica gel
chromatography, eluting with ethyl acetate/hexanes (1:4) to afford
3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(5-1-naphthyl-1H-indol-3-yl) pyrrole-2, 5-dione as
a bright red solid (70 mg, 71%). .sup.1H NMR (CD.sub.3OD) .delta.:
1.80-1.92 (m, 2H), 2.72-2.80 (t, J=6.0 Hz, 2H), 3.94-3.99 (t, J=6.0
Hz, 2H), 6.50-6.58 (m, 3H), 6.66 (s, 1H), 6.72 (m, 1H), 6.98 (dd,
J=8.4 Hz, J'=2.0 Hz, 1H), 7.00-7.50 (m, 2H), 7.28 (dd, J=6.8 Hz,
J'=8.4 Hz, 1H), 7.38-7.43 (m, 2H), 7.61 (s, 1H), 7.72 (d, J=8.4 Hz,
1H), 7.82 (d, J=8.4 Hz, 1H), 7.97 (s, 1H).
Example 32
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-phenyl-1H-indol-3-yl) pyrrole-2, 5-dione
[0259] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-phenyl-1H-indol-3-yl) pyrrole-2, 5-dione was
prepared according to the method of Example 31 employing phenyl
boronic acid in place of 1-naphthyl boronic acid. .sup.1H NMR
(CD.sub.3OD) .delta.: 2.10-2.18 (m, 2H), 2.90 (t, J=5.6 Hz, 2H),
4.18 (t, J=5.6 Hz, 2H), 6.63 (t, J=7.6 Hz, 1H), 6.75-6.83 (m, 5H),
7.11-7.20 (m, 3H), 7.22 (dd, J=8.4 Hz, J'=1.2 Hz, 1H), 7.38 (d,
J=8.4 Hz, 1H), 7.59 (s, 1H), 7.93 (s, 1H).
Example 33
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(4-methoxyphenyl)-1H-indol-3-yl) pyrrole-2,
5-dione
[0260] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(4-methoxyphenyl)-1H-indol-3-yl) pyrrole-2,
5-dione was prepared according to the method of Example 31
employing 4-methoxyphenyl boronic acid in place of 1-naphthyl
boronic acid. .sup.1H NMR (CD.sub.3OD) .delta.: 2.09-2.18 (m, 2H),
2.90 (t, J=6.0 Hz, 2H), 4.15 (t, J=6.0 Hz, 2H), 6.62-6.68 (m, 2H),
6.73 (s, 4H), 6.77-6.82 (m, 2H), 7.18 (d, J=8.4 Hz, 1H), 7.33 (d,
J=8.4 Hz, 1H), 7.53 (s, 1H), 7.91 (s, 1H).
Example 34
Preparation of 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(5-(3-methylphenyl)-1H-indol-3-yl) pyrrole-2,
5-dione
[0261] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(3-methylphenyl)-1H-indol-3-yl) pyrrole-2,
5-dione was prepared according to the method of Example 31
employing 3-methylphenyl boronic acid in place of l-naphthyl
boronic acid. .sup.1H NMR (CD.sub.3OD) .delta.: 2.00-2.10 (m, 2H),
2.11 (s, 3H). 2.81-2.88 (t, J=6.0 Hz, 2H), 4.03-4.11 (t, J=5.6 Hz,
2H), 6.50 (d, J=7.2 Hz, 1H), 6.64 (t, J=7.6 Hz, 1H), 6.74-6.81 (m,
3H), 6.86 (d, J=8.0 Hz, 1H), 6.94 (d, J=7.6 Hz, 1H), 7.03 (t, J=7.2
Hz, 1H), 7.22 (dd, J=8.4 Hz, J'=2.0 Hz, 1H), 7.36 (d, J=8.8 Hz,
1H), 7.48 (s, 1H), 7.90 (s, 1H).
Example 35
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-bromo-1H-indol-3-yl) pyrrolidine-2, 5-dione
[0262] 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-bromo-1-indol-3-yl) pyrrole-2, 5-dione, prepared
as in Example 24, was reduced with Mg in methanol as described in
Example 2, Procedure C, to yield
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-bromo-1H-indol-3-yl) pyrrolidine-2, 5-dione.
.sup.1H NMR (CD.sub.3OD) .delta.: 2.18-2.26 (m, 2H), 2.96 (t, J=6.0
Hz, 2H), 4.12 (t, J=6.4 Hz, 2H), 4.40 (d, J=6.8 Hz, 1H), 4.52 (d,
J=6.8 Hz, 1H), 6.86-6.96 (m, 2H), 7.08 (s, 1H), 7.13-7.30 (m,
5H)
Example 36
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-phenyl-1H-indol-3-yl) pyrrolidine-2, 5-dione
[0263] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-phenyl-1H-indol-3-yl) pyrrole-2, 5-dione,
prepared as in Example 32, was reduced with Mg in methanol as
described in Example 2, Procedure C, to yield
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-phenyl-1H-indol-3-yl) pyrrolidine-2, 5-dione.
.sup.1H NMR (CD.sub.3OD) .delta.: 2.00-2.16 (m, 2H), 2.94 (t, J=6.0
Hz, 2H), 3.92-3.99 (m, 1H), 4.00-4.08 (m, 1H), 4.36 (d, J=6.4 Hz,
1H), 4.68 (d, J=6.4 Hz 1H), 6.88-6.97 (m, 2H), 7.04 (s, 1H),
7.12-7.15 (m, 1H), 7.17-7.47 (m, 9H).
Example 37
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(1-naphthyl)-1H-indol-3-yl) pyrrolidine-2,
5-dione
[0264] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-1-naphthyl-1H-indol-3-yl) pyrrole-2, 5-dione,
prepared as in Example 31, was reduced with Mg in methanol as
described in Example 2, Procedure C, to yield
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(1-naphthyl)-1H-indol-3-yl) pyrrolidine-2,
5-dione. .sup.1H NMR (CD.sub.3OD) .delta.: 1.85-1.95 (m, 1H),
1.95-2.05 (m, 1H), 2.74-2.88 (m, 2H), 3.72-3.83 (m, 1H), 3.88-3.98
(m, 1H), 4.40 (d, J=6.4 Hz, 1H), 4.62 (d, J=6.4 Hz, 1H), 6.80 (d,
J=6.8 Hz, 1H), 6.78 (t, J=8.0 Hz, 1H), 7.46 (s, 1H), 7.07-7.13 (m,
2H), 7.18-7.23 (dd, J=8.4 Hz, J=1.6 Hz, 2H), 7.27-7.34 (m, 2H),
7.41-7.49 (m, 3H), 7.78-7.83 (dd, J=8.4 Hz, J=3.2 Hz, 2H),
7.86-7.90 (d, J=7.6 Hz, 1H).
Example 38
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(4-methoxyphenyl)-1H-indol-3-yl) pyrrolidine-2,
5-dione
[0265] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(4-methoxyphenyl)-1H-indol-3-yl) pyrrole-2,
5-dione, prepared as in Example 33, was reduced with Mg in methanol
as described in Example 2, Procedure C, to yield
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(4-methoxyphenyl)-1H-indol-3-yl) pyrrolidine-2,
5-dione. .sup.1H NMR (CD.sub.3OD) .delta.: 2.03-2.22 (m, 2H), 2.98
(t, J=6.0 Hz, 2H), 3.80 (s, 3H), 3.97-4.06 (m, 1H), 4.06-4.14 (m,
1H), 4.38 (d, J=6.8 Hz, 1H), 4.67 (d, J=6.8 Hz, 1H), 6.86 (d, J=8.4
Hz, 2H), 6.91-7.00 (m, 2H), 7.08 (s, 2H), 7.17-7.27 (m, 4H), 7.31
(d, J=8.4 Hz, 1H), 7.37 (d, J=8.8 Hz, 1H).
Example 39
(.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(3-methylphenyl)-1H-indol-3-yl) pyrrolidine-2,
5-dione
[0266] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(3-methylphenyl)-1H-indol-3-yl) pyrrole-2,
5-dione, prepared as in Example 34, was reduced with Mg in methanol
as described in Example 2, Procedure C, to yield
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(5-(3-methylphenyl)-1H-indol-3-yl) pyrrolidine-2,
5-dione. .sup.1H NMR (CD.sub.3OD) .delta.: 1.98-2.18 (m, 2H), 2.34
(s, 3H), 2.85-3.00 (m, 2H), 3.90-3.98 (m, 1H), 3.98-4.09 (m, 1H),
4.35 (d, J=7.2 Hz, 1H), 4.64 (d, J=6.8 Hz, 1H), 6.88-6.99 (m, 2H),
7.00-7.10 (m, 3H), 7.13-7.26 (m, 5H), 7.36 (m, 2H).
Example 40
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2-chloro-4-fluorophenyl) pyrrolidine-2,
5-dione
[0267] To a solution of 5, 6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl) oxoacetic acid methyl ester (0.243 g, 1 mmol) and
2-chloro-4-fluorophenylacetamide (1 mmol) in anhydrous
tetrahydrofuran (5 ml) at 0.degree. C. was added a solution of
potassium t-butoxide (1 M in tetrahydrofuran) (2.5 ml, 2.5 mmol).
The mixture was stirred at 0.degree. C. for 2 hours. Concentrated
hydrochloric acid (0.5 ml) was then added and the mixture stirred
for 1 hour at room temperature. The mixture was then diluted with
ethyl acetate (20 ml), washed with water (2.times.15 ml) and
saturated aqueous sodium chloride solution (15 ml). The organic
layer was then dried over anhydrous sodium sulfate and concentrated
under reduced pressure to yield an oil. This residue was diluted in
anhydrous methanol (15 ml) and the resulting solution charged with
oven dried magnesium turnings (0.5 g, 20.5 mmol) and stirred at
70.degree. C. in a ventilated vial until the Mg turnings fully
dissolved or for two hours. The vial was then allowed to cool to
room temperature. The mixture was diluted with ethyl acetate (25
ml) and washed with 10% hydrochloric acid (2.times.25 m) and
saturated aqueous sodium chloride solution (20 ml). The organic
layer was dried over anhydrous sodium sulfate and concentrated
under reduced pressure. The residue was purified by silica gel
chromatography, eluting with an ethyl acetate/hexanes gradient (10%
ethyl acetate to 50% ethyl acetate over 40 minutes) to yield (25.6
mg, 6.7%) of (.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2-chloro-4-fluorophenyl) pyrrolidine-2, 5-dione.
.sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 12.5 (s, 1H), 7.52 (t,
1H, J=6.4 Hz), 7.49 (dd, 1H, J=6.4 2.4 Hz), 7.34 (s, 1H), 7.21 (td,
1H, J=6.0 2.8 Hz), 7.10 (d, 1H, J=7.6 Hz), 6.87 (m, 2H), 4.67 (d,
1H, J=8.0 Hz), 4.51 (d, 1H, J=7.2 Hz), 2.90 (t, 2H, J=5.6 Hz), 2.11
(t, 2H, J=5.2 Hz).
Example 41
Preparation of (.+-.+)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2,6-dichlorophenyl) pyrrolidine-2, 5-dione
[0268] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2,6-dichlorophenyl) pyrrolidine-2, 5-dione was
prepared according to Example 40 replacing
2-chloro-4-fluorophenylacetamide with 2,6-dichlorophenylacetamide.
Yield 52.2 mg, 13.0%. .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.:
11.82 (s, 1H), 7.34 (m, 3H), 7.10 (d, 1H, J=7.2 Hz), 6.87 (m, 2H),
5.16 (d, 1H J=7.6 Hz), 5.10 (d, 1H, J=7.6 Hz), 2.91 (t, 2H, J=6.0
Hz) 2.10 (m, 2H).
Example 42
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(4-bromophenyl) pyrrolidine-2, 5-dione
[0269] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(4-bromophenyl) pyrrolidine-2, 5-dione was prepared
according to Example 40 replacing 2-chloro-4-fluorophenylacetamide
with 4-bromophenylacetamide. Yield 33.1 mg, 8.1%. .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 11.55 (s, 1H), 7.53 (dt, 2H, J=8.8
2.0 Hz), 7.34 (dt, 3H, J=8.0 2.0 Hz), 7.15 (dd, 1H, J=7.6 1.0 Hz),
6.86 (m, 2H), 4.53 (d, 1H, J=8.0 Hz), 4.37 (d, 1H, J=8.0 Hz), 4.10
(t, 2H, J=1.6 Hz), 2.90 (t, 2H, J=2.0 Hz), 2.12 (t, 2H, J=1.8
Hz).
Example 43
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(4-chlorophenyl) pyrrolidine-2, 5-dione
[0270] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(4-chlorophenyl) pyrrolidine-2, 5-dione was
prepared according to Example 40 replacing
2-chloro-4-fluorophenylacetamide with 4-chlorophenylacetamide.
Yield 32.7 mg, 9.0%. .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.:
11.54 (s, 1H), 7.40 (m, 4H), 7.33 (s, 1H), 7.15 (dd, 1H, J=6.8 0.8
Hz), 6.86 (m, 2H), 4.54 (d, 1H, J=8.0 Hz), 7.38 (d, 1H, J=7.6 Hz),
4.10 (t, 2H, J=5.6 Hz), 2.90 (t, 2H, J=6.0 Hz), 2.11 (m, 2H).
Example 44
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(4-trifluoromethoxyphenyl) pyrrolidine-2,
5-dione
[0271] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(4-trifluoromethoxyphenyl) pyrrolidine-2, 5-dione
was prepared according to Example 40 replacing
2-chloro-4-fluorophenylacetamide with
4-trifluoromethoxyphenylacetamide. Yield 67.8 mg, 16.4%. .sup.1H
NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.56 (s, 1H), 7.52 (d, 2H,
J=8.4 Hz), 7.35 (s, 1H), 7.33 (d, 2H, J=8.0 Hz), 7.15 (d, 1H, J=7.2
Hz), 6.86 (m, 2H), 4.58 (d, 1H, J=8.0 Hz), 4.45 (d, 1H, J=8.0 Hz),
4.10 (t, 2H, J=6.0 Hz), 2.90 (t, 2H, J=6.0), 2.10 (t, 2H,
J=5.6).
Example 45
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(thiophen-3-yl) pyrrolidine-2, 5-dione
[0272] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(thiophen-3-yl) pyrrolidine-2, 5-dione was prepared
according to Example 40 replacing 2-chloro-4-fluorophenylacetamide
with thiophen-3-ylacetamide. Yield 50.3 mg, 15.0%. .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 11.50 (s, 1H), 7.52 (m, 1H), 7.49
(m, 1H), 7.35 (s, 1H), 7.21 (dd, 1H, J=4.0 1.2 Hz), 7.16 (d, 1H,
7.6 Hz), 6.89 (d, 1H, J=4.4 Hz), 6.85 (t, 1H, J=6.8 Hz), 4.56 (d,
1H, J=7.2 Hz), 4.41 (d, 1H, J=7.2 Hz), 4.10 (t, 2H, J=6.0 Hz), 2.90
(t, 2H, J=6.0 Hz), 2.10 (m, 2H).
Example 46
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2-fluorophenyl) pyrrolidine-2, 5-dione
[0273] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2-fluorophenyl) pyrrolidine-2, 5-dione was
prepared according to Example 40 replacing
2-chloro-4-fluorophenylacetamide with 2-fluorophenylacetamide.
Yield 30.6 mg, 8.8%. .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.:
11.64 (s, 1H), 7.36 (m, 3H), 7.17 (m, 3H), 6.84 (m, 2H), 4.44 (d,
1H, J=7.2 Hz), 4.40 (d, 1H, J=7.6 Hz), 4.10 (s, 2H), 2.88 (s, 2H),
2.09 (s, 2H).
Example 47
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2-thiophen-2-yl) pyrrolidine-2, 5-dione
[0274] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2-thiophen-2-yl) pyrrolidine-2, 5-dione was
prepared according to Example 40 replacing
2-chloro-4-fluorophenylacetamide with 2-thiophen-2-ylacetamide.
Yield 30.6 mg, 8.8%. .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.:
11.58 (s, 1H), 7.45 (dd, 1H, J=5.2 0.8 Hz), 7.40 (s, 1H), 7.22 (d,
1H, J=8.0 Hz), 7.12 (d, 1H, J=3.2 Hz), 6.99 (dd, 1H, J=5.2 and 3.6
Hz), 4.63 (d, 1H, J=8.0 Hz), 4.60 (d, 1H, J=7.6 Hz), 2.90 (t, 2H,
J=6.0 Hz), 2.12 (t, 2H, J=6.0 Hz).
Example 48
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2,4-dichlorophenyl pyrrolidine-2, 5-dione
[0275] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2,4-dichlorophenyl) pyrrolidine-2, 5-dione was
prepared according to Example 40 replacing
2-chloro-4-fluorophenylacetamide with 2,4-dichlorophenylacetamide.
Yield 20.9 mg, 5.2%. .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.:
11.65 (s, 1H), 7.69 (s, 1H), 7.51 (d, 1H, J=8.0 Hz), 7.43 (d, 1H,
J=8.0 Hz), 7.34 (s, 1H), 7.12 (m, 1H), 6.87 (m, 2H), 4.65 (d, 11H,
J=7.6 Hz), 4.55 (d, 1H, J=7.6 Hz), 4.10 (t, 2H, J=6.0 Hz), 2.90 (t,
2H, J=6.0), 2.12 (t, 2H, J=6.0 Hz).
Example 49
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-phenyl-pyrrolidine-2, 5-dione
[0276] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-phenyl-pyrrolidine-2, 5-dione was prepared
according to the Example 40 replacing
2-chloro-4-fluorophenylacetamide with phenylacetamide. .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 11.511 (s, 1H), 7.24-7.36 (m, 6H),
7.13 (d, 1H, J=7.2), 6.8-6.88 (m, 2H), 4.49 (d, 1H, J=8.0 Hz), 4.3
(d, 1H, J=7.6 Hz), 4.08 (m, 2H), 2.88 (m, 2H), 2.088 (m, 2H).
Example 50
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2-chlorophenyl)-pyrrolidine-2, 5-dione
[0277] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2-chlorophenyl)-pyrrolidine-2, 5-dione was
prepared according to Example 40 replacing
2-chloro-4-fluorophenylacetamide with 2-chlorophenylacetamide.
.sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.655 (s, 1H),
7.41-7.48 (m, 2H), 7.27-7.35 (m, 3H, J=7.2), 7.87 (d, 1H, J=7.6),
6.81-6.88 (m, 2H), 4.632 (d, 1H, J=7.6 Hz), 4.494 (d, 1H, J=7.2),
4.07-4.10 (m, 2H), 2.884 (m, 2H), 2.09 (m, 2H).
Example 51
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(N-methylindol-3-yl) pyrrolidine-2, 5-dione
[0278] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(N-methylindol-3-yl) pyrrolidine-2, 5-dione was
prepared according to Example 40 replacing
2-chloro-4-fluorophenylacetamide with N-methylindol-3-ylacetamide.
.sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.55 (s, 1H),
7.44-7.34 (m, 4H), 7.2-7.18 (m, 2H), 7.01 (t, 1H), 6.82-6.89 (m,
2H), 4.49 (dd, 2H), 4.093 (t, 2H), 4.093 (t, 2H), 3.73 (s, 3H),
2.89 (t, 2H), 2.07 (m, 2H).
Example 52
Preparation of (.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(4-methoxyphenyl)-pyrrolidine-2, 5-dione
[0279] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(4-methoxy-phenyl)-pyrrole-2, 5-dione, prepared as
in Example 10 and was reduced by employing the method of Example 2,
Protocol 13, to yield (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1yl)-4-(4-methoxyphenyl)-pyrrolidine-2,
5-dione. .sup.1H-NMR (CDCl.sub.3) 400 MHz .delta.: 8.62 (s, 1H),
7.15 (d, 1H, J=7.6 Hz), 6.8-6.93 (m, 4H), 6.7 (s, 1H), 6.55 (d, 2H,
J=8.4 Hz), 4.8 (d, 1H, J=8.8 Hz), 4.48 (d, 1H, J=8.8 Hz), 3.96 (m,
2H), 3.63 (s, 3H), 2.87 (t, 2H, J=6 Hz), 2.10 (m, 2H).
Example 53
Preparation of (.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2,5-dimethoxyphenyl)-pyrrolidine-2, 5-dione
[0280] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2,5-dimethoxy-phenyl)-pyrrole-2, 5-dione,
prepared as in Example 17, was reduced by employing the method of
Example 2, Protocol B, to yield
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2,5-dimethoxyphenyl)-pyrrolidine-2, 5-dione.
.sup.1H NMR (CDCl.sub.3) 400 MHz .delta.: 8.0 (s, 1H), 7.19 (d, 1H,
J=7.6 Hz), 6.89 (t, 1H, J=7.2 Hz), 6.77 (d, 2H, J=7.2 Hz),
6.44-6.51 (m, 3H), 4.84 (d, 2H, J=9.6 Hz), 3.88-4.00 (m, 2H), 3.6
(s, 3H), 3.49 (s, 3H), 2.8 (m, 2H), 2.05 (m, 2H).
Example 54
Preparation of (.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2-chloro-4-fluoro-phenyl)-pyrrolidine-2,
5-dione
[0281] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(2-chloro-4-fluoro-phenyl)-pyrrole-2, 5-dione,
prepared as in Example 18, was reduced by employing the method of
Example 2, Protocol B, to yield
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(2-chloro-4-fluoro-phenyl)-pyrrolidine-2, 5-dione.
.sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.82 (s, 1H),
7.02-7.18 (m, 4H), 6.7-6.85 (m, 3H), 5.01 (d, 1H, J=9.2 Hz), 4.79
(d, 2H, J=9.6 Hz), 3.96 (m, 2H), 2.79 (m, 2H), 1.97 (m, 2H).
Example 55
Preparation of (.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4-(3-chlorophenyl)-pyrrolidine-2, 5-dione
[0282] 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4-(3-chloro-phenyl)-pyrrole-2, 5-dione, prepared as
in Example 15, was reduced by employing the method of Example 2,
Procedure B, to yield (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1yl)-4-(3-chlorophenyl)-pyrrolidine-2, 5-dione.
.sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.66 (s, 1H), 7.13 (d,
1H, J=8 Hz), 6.95-7.02 (m, 5H), 6.78 (t, 1H, J=7.6 Hz), 6.7 (d, 1H,
J=7.2 Hz), 4.84 (d, 1H, J=9.2 Hz), 4.65 (d, 2H, J=8.8 Hz), 3.9-4.03
(m, 2H), 2.79 (t, 2H, J=5.6 Hz), 1.97 (m, 2H).
Example 56
Preparation of (.+-.)-Phosphoric acid
mono-[trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-
-3-yl)-2,5-dioxo-pyrrolidin-1-ylmethyl] ester
Step 1
##STR00020##
[0284] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione (3.0 g, 8.13
mmol, prepared as in Example 2, Procedure C) and formaldehyde (30
ml, 37% in water) in tetrahydrofuran (30 ml) were stirred for 14-16
hours at room temperature. The mixture was then taken up in ethyl
acetate (50 ml) and water (50 ml). The organic layer was washed
with brine and dried over sodium sulfate. Solvent was removed under
reduced pressure and residue was purified using a silica gel
chromatography column eluted with EtOAc/Hexane 1:1 to yield 2.5 g,
77%, of (.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij]quinolin-1-yl)-1-hydroxymethyl-4-(1H-indol-3-yl)-pyrrolidine-2,-
5-dione an orange foamy solid (2.5 g, 77%).
Step 2
##STR00021##
[0286]
(.+-.)-Trans-3-(5,6-Dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-1-hy-
droxymethyl-4-(1H-indol-3-yl)-pyrrolidine-2,5-dione (0.06 g) in
anhydrous tetrahydrofuran (5 ml) was treated with
dibenzylphosphoramidate (0.156 ml, 3.5 equivalents) followed by the
addition of tetrazole (3% solution in acetonitrile, 2 ml). The
reaction mixture was stirred at room temperature for 20 min and
cooled to -78.degree. C. A solution of m-chloroperbenzoic acid
(70%, 0.162 g) in dichloromethane (2 ml) was added at -78.degree.
C. After 5 min at -78.degree. C., the reaction was brought to room
temperature and stirred for 5 min. Solvents were removed under
reduced pressure and the residue was purified by flash
chromatography on a silica column, eluted with ethyl acetate,
hexane to give phosphoric acid dibenzyl ester
trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-yl)-4-(1H-indol-3-yl)-2-
,5-dioxo-pyrrolidin-1-ylmethyl ester as a solid (70 mg). .sup.1H
NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.10 (s, 1H), 7.32-7.39 (m,
12H), 6.84-7.24 (m, 2H), 5.49 (brs, 2H), 5.03 (m, 4H), 4.61 (dd,
2H), 4.06 (brs, 2H), 2.87 (brs, 2H), 2.07 (brs, 2H).
Step 3
##STR00022##
[0288] The phosphoric acid dibenzyl ester of
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indo-
l-3-yl)-2,5-dioxo-pyrrolidin-1-yl-methyl ester (0.160 g) in
methanol (2 ml) and Pd/C (10%, 20 mg) was stirred at room
temperature under 1 atmosphere of hydrogen for two hours. The
mixture was filtered over Celite and the solvent removed to give
(.+-.)-phosphoric acid
mono-[trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-
-3-yl)-2,5-dioxo-pyrrolidin-1-ylmethyl] ester (0.110 g).
Example 57
Preparation of (.+-.)-trans-2-Amino-propionic
acid-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)--
2,5-dioxo-pyrrolidin-1-yl methyl ester
Step 1
##STR00023##
[0290] To a solution of
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-1-hydroxym-
ethyl-4-(1H-indol-3-yl)-pyrrolidine-2,5-dione (0.5 mmol) in
tetrahydrofuran (8 ml) was added N-carbobenzyloxy alanine (1.1
equivalents) followed by the addition of HBTU (1.5 equivalents) and
DIPEA (2.2 equivalents). The mixture was stirred at room
temperature for 15 h. The solvents were removed under reduced
pressure and the residue was taken up in ethyl acetate and water
(1:1, 15 ml). The organic layer was separated and dried. The
residue was purified by silica gel chromatography to provide the
N-carbobenzyloxy protected product.
Step 2
##STR00024##
[0292] A solution of the N-carbobenzyloxy protected product from
Step 1 (0.5 mmol) in methanol (8 ml) and a few drops of 4 M HCl in
ethyl acetate and 10% Pd/C (10% w/w) were stirred at room
temperature under 1 atmosphere of hydrogen for 2 hours. The mixture
was then filtered over celite and the solvent removed to provide
final product (.+-.)-trans-2-amino-propionic
acid-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)--
2,5-dioxo-pyrrolidin-1-ylmethyl ester. .sup.1H NMR (DMSO-d.sub.6)
400 MHz .delta.: 11.10 (s, 11H), 8.57 (s, 2H), 6.84-7.41 (m, 9H),
5.61 (m, 2H), 4.62 (dd, 2H), 4.07 (brs, 2H), 3.72 (brm, 1H), 2.87
(brs, 2H), 2.23 (s, 6H), 2.08 (brs, 2H), 1.40 (d, J=6.4 Hz,
3H).
Example 58
Preparation of (.+-.)-trans-2-Amino-acetic
acid-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)--
2,5-dioxo-pyrrolidin-1-yl methyl ester
[0293] (.+-.)-trans-2-Amino-acetic
acid-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)--
2,5-dioxo-pyrrolidin-1-ylmethyl ester was prepared as in Example 57
by replacing N-carbobenzyloxy alanine with N-carbobenzyloxy
glycine. .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.19 (s, 1H),
8.46 (s, 2H), 6.82-7.43 (m, 9H), 5.61 (s, 2H), 4.65 (dd, 2H), 4.08
(brt, J=5.6 Hz, 2H), 3.88 (brs, 2H), 2.87 (t, J=5.6 Hz, 2H), 2.48
(s, 2H), 2.08 (t, J=4.8 Hz, 2H).
Example 59
Preparation of (.+-.)-trans-2-dimethylamino-acetic
acid-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)--
2,5-dioxo-pyrrolidin-ylmethyl ester
[0294] To a solution of
(.+-.)-trans-3-(5,6-Dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-1-hydroxym-
ethyl-4-(1H-indol-3-yl)-pyrrolidine-2,5-dione (0.5 mmol) in
tetrahydrofuran (8 ml) was added N,N-dimethylglycine (1.1
equivalents) followed by the addition of HBTU (1.5 equivalents) and
DIPEA (N,N-diisopropylethylamine, 2.2 equivalents). The mixture was
stirred at room temperature for 15 hours. The solvents were removed
under reduced pressure and the residue was taken up in ethyl
acetate and water (1:1, 15 ml). The organic layer was separated and
dried to yield a residue. The residue was purified by
chromatography on a silica gel column eluted with ethyl acetate
hexanes to yield (.+-.)-trans-2-dimethylamino-acetic
acid-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)--
2,5-dioxo-pyrrolidin-1-yl methyl ester. .sup.1H NMR (DMSO-d.sub.6)
400 MHz .delta.: 11.10 (s, 1H), 6.82-7.41 (m, 9H), 5.70 (m, 2H),
4.62 (dd, 2H), 4.07 (brs, 2H), 3.23 (s, 2H), 2.87 (brs, 2H), 2.23
(s, 6H), 2.08 (brs, 2H).
Example 60
Preparation of (.+-.)-trans-Isonicotinic
acid-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)--
2,5-dioxo-pyrrolidin-1-ylmethyl ester
[0295] (.+-.)-trans-Isonicotinic acid
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)-2,5-d-
ioxo-pyrrolidin-1-ylmethyl ester was prepared as in Example 59 by
replacing N,N-dimethylglycine with 4-carboxypyridine. .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 11.19 (s, 1H), 8.83 (d, 2H), 7.83
(d, 2H), 6.83-7.42 (m, 9H), 5.88 (s, 2H), 4.65 (dd, 2H), 4.05 (brt,
2H), 2.86 (brs, 2H), 2.08 (brs, 2H).
Example 61
Preparation of 3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1-methylindol-3-yl)-1-methyl pyrrole-2, 5-dione
and (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1-methylindol-3-yl)-1-methyl pyrrolidine-2,
5-dione
[0296] Step 1: To a solution of 3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrole-2, 5-dione (100
mg, see Example 1) in anhydrous dimethylformamide (5 ml) was added
potassium carbonate (0.5 g) and methyl iodide (0.1 ml). The mixture
was stirred at room temperature for 48 hours then poured into ethyl
acetate (100 ml), washed with water (100 ml), dried over anhydrous
sodium sulfate and evaporated to give 3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-methylindol-3-yl)-1-methyl
pyrrole-2, 5-dione as a red solid (93 mg).
[0297] Step 2: To a solution of 3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1-methylindol-3-yl)-1-methyl pyrrole-2,
5-dione (93 mg) in methanol (5 ml) and ethylacetate (5 ml) was
added 10% Pd--C (50 mg) and the mixture stirred at room temperature
under 1 atmosphere of hydrogen for 48 hours. Toluene (50 ml) was
added and the mixture again stirred at room temperature under 1
atmosphere of hydrogen for 2 hours. The mixture was then filtered
through a pad of celite and evaporated to dryness to yield a
residue. The residue was purified using silica gel chromatography
eluting with 35-40% ethylacetate in hexanes to give
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1-methylindol-3-yl)-1-methyl pyrrolidine-2,
5-dione as a pale yellow solid (53 mg). .sup.1H NMR (CDCl.sub.3)
400 MHz .delta.: 7.23 (s, 1H), 7.05-7.07 (m, 2H), 7.01 (d, 1H,
J=7.2 Hz), 6.92-6.97 (m, 1H), 6.85 (t, 1H, J=7.2 Hz), 6.74 (d, 1H,
J=6.8 Hz), 6.64 (d, 2H, J=6.4 Hz), 4.78 (m, 2H), 3.75-3.84 (m, 2H),
3.45 (s, 3H), 3.27 (s, 3H), 2.79 (t, 2H, J=5.6 Hz), 1.98 (m,
2H).
Example 62
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
[0298] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione may be
prepared by reacting 1H-indole and
3,4-dibromo-1-phenyl-pyrrole-2,5-dione in the presence of methyl
magnesium bromide to yield
3-bromo-4-(1H-indol-3-yl)-1-phenyl-pyrrole-2,5-dione. The
3-bromo-4-(1H-indol-3-yl)-1-phenyl-pyrrole-2,5-dione is
subsequently reacted with 5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinoline
and LiHMDS (lithium hexamethyldisilane) in toluene or
(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-boranediol and
Pd(PPh.sub.3).sub.4 (tetrakis(triphenylphosphine)palladium) to
yield
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)-1-phe-
nyl-pyrrole-2,5-dione, which is reduced and deprotected by
treatment with Mg in methanol, as in Example 2 procedure C,
followed by catalytic hydrogenation over palladium on carbon to
yield (.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione. Bnz is
benzyl.
##STR00025##
Example 63
Preparation of (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
[0299] (.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione may be
prepared by reacting 1-allyl-7-bromo-1H-indole with (COCl).sub.2
(oxalyl chloride) and sodium methoxide in a polar aprotic solvent
such as dichloromethane to yield
(1-allyl-7-bromo-1H-indol-3-yl)-oxo-acetic acid methyl ester, which
is subsequently reacted with 2-(1H-indol-3-yl)-acetamide and tBuOK
(potassium tert-butoxide) in THF to yield
3-(1-allyl-7-bromo-1H-indol-3-yl)-4-(1H-indol-3-yl)-pyrrole-2,5-dione.
Reduction of the 3-(1-allyl-7-bromo-1
II-indol-3-yl)-4-(1H-indol-3-yl)-pyrrole-2,5-dione by Mg in
refluxing methanol, as in Example 2 procedure C, yields
3-(1-allyl-7-bromo-1H-indol-3-yl)-4-(1H-indol-3-yl)-pyrrolidine-2,5-dione-
, which is treated with 9-BBN (9-borabicyclo[3.3.1]nonane) and
Pd(PPh.sub.3).sub.4 (tetrakis(triphenylphosphine)palladium) to
yield (.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione.
##STR00026##
Example 64
Preparation of (.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione and
(.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
[0300] The cis and trans isomers of 3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
may be prepared beginning with the reaction of
(1H-indol-3-yl)-oxo-acetic acid methyl ester and
(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-acetic acid methyl
ester in the presence of a base such as LDA (lithium
diisopropylamide) in a polar aprotic solvent such as THF to yield
2-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-3-(1H-indol-3-yl)-but-2-
-enedioic acid dimethyl ester. Alternatively,
2-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-3-(1H-indol-3-yl)-but-2-
-enedioic acid dimethyl ester may be prepared by reaction of
(1H-indol-3-yl)-acetic acid methyl ester and
(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-oxo-acetic acid
methyl ester in the presence of a base (e.g., LDA) in THF. The
2-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij]quinolin-1-yl)-3-(1H-indol-3-yl)-but-2-enedioic acid
dimethyl ester is reduced by catalytic hydrogenation over a noble
metal catalyst (e.g., Pd on charcoal) to give
2-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-3-(1H-indol-3-yl)-succi-
nic acid dimethyl ester, which is reacted with benzylamine
(PhCH.sub.2NH.sub.2) to yield a mixture of cis and trans
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)-1-phe-
nyl-pyrrolidine-2,5-dione. The mixture of cis and trans isomers may
be deprotected by catalytic hydrogenation over Pd on charcoal
(Pd--C) to give rise to a mixture of cis and trans
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)-pyrro-
lidine-2,5-dione. The cis and trans isomers may be separated to
give all four cis and trans isomers (e.g., by chromatography as in
Examples 4 and 5). The deprotected mixture of cis and trans isomers
may be treated with potassium tert-butoxide in tert-butanol (as in
Example 3) or a mixture of THF and tert-butanol at 50.degree. C. to
yield a mixture with a predominance of the trans isomers.
Alternatively, the
2-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-3-(1H-indol-3-yl)-succi-
nic acid dimethyl ester can be reacted with ammonia in methanol at
elevated temperatures to yield predominantly the cis isomers of
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)-pyrro-
lidine-2,5-dione, which may be isomerized to yield predominately
the trans isomers of
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(1H-indol-3-yl)-pyrro-
lidine-2,5-dione with potassium tert-butoxide in tert-butanol (as
in Example 3) or a mixture of THF and tert-butanol at 50.degree.
C.
##STR00027##
Example 65
Preparation of
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(3-methoxyphenyl)-1H--
pyrrole-2,5-dione
[0301] To a mixture of 5, 6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl) oxoacetic acid methyl ester (0.50 g, 2.05 mmol) and
2-(3-methoxyphenyl)acetamide (0.37 g, 2.26 mmol) in anhydrous
tetrahydrofuran (5 mL) was added potassium tert-butoxide (1.0 M in
tetrahydrofuran, 6.17 mL, 6.17 mmol) dropwise at 0.degree. C. The
mixture was stirred at 0.degree. C. for 3 hours then concentrated
hydrochloric acid (1.5 mL) was added at 0.degree. C. The resulting
mixture was stirred for 1 hour, diluted with ethyl acetate (150
mL), washed with water (2.times.50 mL), dried over anhydrous sodium
sulfate and concentrated to give 0.91 g of an orange solid. The
residue was purified by column chromatography eluting with 20-40%
ethyl acetate in hexane to give
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(3-methoxyphenyl)-1H--
pyrrole-2,5-dione. Mp 99-101.degree. C.; .sup.1H NMR (CDCl.sub.3)
400 MHz .delta.: 8.01 (s, 1H), 7.81 (bs, 1H), 7.20-7.25 (m, 1H),
7.06-7.08 (min, 2H), 6.85-6.91 (m, 2H), 7.25 (t, 1H), 6.13 (d,
J=8.0 Hz, 1H), 4.24 (t, 2H), 3.66 (s, 3H), 2.97 (t, J=6.0 Hz, 2H),
2.22-2.26 (m, 2H).
Example 66
Preparation of
3-[4-(benzyloxy)phenyl]-4-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-
-1H-pyrrole-2,5-dione
[0302]
3-[4-(benzyloxy)phenyl]-4-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-
-1-yl)-1H-pyrrole-2,5-dione was prepared according to Example 65,
employing 2-(4-(benzyloxy)phenyl)acetamide in place of
2-(3-methoxyphenyl)acetamide. Mp 262-265.degree. C.; .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 10.96 (s, 1H), 8.01 (d, 1H),
7.33-7.45 (m, 7H), 6.99 (d, J=6.8 Hz, 2H), 6.83 (d, J=7.2 Hz, 1H),
6.63 (t, 1H), 6.09 (d, J=8.4 Hz, 1H), 5.13 (s, 2H), 4.27 (m, 2H),
2.92 (m, 2H), 2.15 (m, 2H).
Example 67
Preparation of
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(4-fluorophenyl)-1H-p-
yrrole-2,5-dione
[0303]
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(4-fluorophenyl-
)-1H-pyrrole-2,5-dione was prepared according to Example 65,
employing 2-(4-fluorophenyl)acetamide in place of
2-(3-methoxyphenyl)acetamide. Mp 234-235.degree. C.; .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 11.05 (s, 1H), 8.07 (d, 1H),
7.42-7.46 (m, 2H), 7.71-7.22 (m, 2H), 6.83 (d, J=7.2 Hz, 1H),
6.65-6.69 (m, 1H), 6.00 (d, J=8.0 Hz, 1H), 4.21 (s, 2H), 2.92 (bs,
2H), 2.15 (bs, 2H).
Example 68
Preparation of
(.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(3-metho-
xyphenyl)pyrrolidine-2,5-dione
[0304] A mixture of
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(3-methoxyphenyl)-1H--
pyrrole-2,5-dione (0.73 g, 2.04 mmol), magnesium (0.89 g, 36.7
mmol) in anhydrous methanol was heated to reflux for 1.5 h. After
cooling to room temperature, the light yellow solution was diluted
with ethyl acetate (200 mL), washed with 1.0 M hydrochloric acid
(2.times.50 mL), water (100 mL), dried over sodium sulfate and
concentrated to provide a light brown solid. The residue was
purified by column chromatography on silica gel eluting with 40-50%
ethyl acetate in hexane to yield
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(3-metho-
xyphenyl)pyrrolidine-2,5-dione as a light yellow solid. Mp
87-91.degree. C.; .sup.1H NMR (CDCl.sub.3) 400 MHz .delta.: 8.73
(s, 1H), 7.25-7.30 (m, 1H), 7.14 (d, J=7.6 Hz, 1H), 6.93-7.01 (m,
3H), 6.77-6.86 (m, 3H), 4.36 (d, J=6.4 Hz, 1H), 4.24 (d, J=6.4 Hz,
1H), 4.18 (t J=5.5 Hz, 2H), 3.78 (s, 3H), 2.97 (t, J=5.6 Hz, 2H),
2.19-2.24 (m, 2H).
Example 69
Preparation of
(.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(3-hydro-
xyphenyl)pyrrolidine-2,5-dione
[0305] To a solution of
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(3-metho-
xyphenyl)pyrrolidine-2,5-dione in dichloromethane (10 mL) at
-78.degree. C. under an atmosphere of nitrogen was slowly added
boron tribromide (1.0 M in dichloromethane) (5.2 mL). The resulting
mixture was stirred at -78.degree. C. for 30 minutes and at room
temperature for 3 hours. The reaction mixture was cooled to
-78.degree. C. then quenched by the addition of methanol (5 mL).
The mixture was allowed to warm to room temperature and maintained
at room temperature for 30 minutes. The reaction mixture was
diluted with dichloromethane (80 mL), washed with saturated aqueous
sodium bicarbonate (15 mL), water (15 mL) and saturated sodium
chloride (15 mL). The organic layer was dried over anhydrous sodium
sulfate and concentrated to dryness. The residue was purified by
flash chromatography on silica gel eluting with ethyl
acetate:hexane:dichloromethane (5:5:1, v/v) to give
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(3-hydro-
xyphenyl)pyrrolidine-2,5-dione as a brown solid (1.15 g, 63%); Mp
108-110.degree. C. 1H NMR (CDCl3) 400 MHz .delta.: 8.69 (s, 1H),
7.18 (t, 1H, J=8.0 Hz), 7.12 (d, 1H, J=8.0 Hz), 6.99 (d, 1H, J=6.8
Hz), 6.97 (d, 1H, J=2.0 Hz), 6.93 (d, 1H, J=6.4 Hz), 6.76 (m, 1H),
6.69 (d, 1H, J=1.6 Hz), 5.67 (brs, 1H), 4.32 (d, 1H, J=6.0 Hz),
4.20 (d, 1H, J=6.0 Hz), 4.07 (t, 2H, J=5.6 Hz), 2.96 (t, 2H, J=6.0
Hz), 2.19 (m, 2H), LC/MS: 347.3 [M+H].
Example 70
Preparation of
(.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(4-fluor-
ophenyl)pyrrolidine-2,5-dione
[0306]
3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(4-fluorophenyl-
)-1H-pyrrole-2,5-dione prepared according to Example 67, was
reduced by employing the method of Example 68 to yield
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(4-fluor-
ophenyl)pyrrolidine-2,5-dione
1-ij]quinolin-1-yl)-4-(4-fluorophenyl)-1H-pyrrole-2,5-dione. Mp
208-210.degree. C.; .sup.1H NMR (DMSO-dc) 400 MHz .delta.: 11.52
(s, 1H), 7.40-7.43 (m, 2H), 7.32 (m, 1H), 7.13-7.17 (m, 3H),
6.82-6.89 (m, 2H), 4.53 (m, 1H), 4.36 (m, 1H), 4.09 (t, J=5.2 Hz,
2H), 2.89 (t, J=6.0 Hz, 2H), 2.09-2.11 (m, 2H).
Example 71
Preparation of
(.+-.)-Trans-3-[4-(benzyloxy)phenyl]-4-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]q-
uinolin-1-yl)pyrrolidine-2,5-dione
[0307]
3-[4-(benzyloxy)phenyl]-4-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-
-1-yl)-1H-pyrrole-2,5-dione prepared according to Example 66, was
reduced by employing the method of Example 68 to yield
(.gamma.)-trans
3-[4-(benzyloxy)phenyl]-4-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-
pyrrolidine-2,5-dione. Mp 91-93.degree. C.; .sup.1H NMR
(DMSO-d.sub.6) 400 MHz .delta.: 11.47 (s, 1H), 7.25-7.43 (m, 8H),
7.15 (d, J=7.6 Hz, 2H), 6.82-6.96 (m, 4H), 5.07 (s, 2H), 4.45 (d,
J=7.6 Hz, 1H), 4.24 (d, J=7.6 Hz, 1H), 4.07-4.10 (m, 2H), 2.87-2.90
(m, 2H), 2.09-2.10 (m, 2H).
Example 72
Preparation of
(.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(4-hydro-
xyphenyl)pyrrolidine-2,5-dione
[0308] A mixture of
(.+-.)-trans-3-[4-(benzyloxy)phenyl]-4-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]q-
uinolin-1-yl)pyrrolidine-2,5-dione (0.2 g) and Pd/C (10% w/w, 0.076
g) was stirred under 1 atmosphere of hydrogen gas overnight. The
catalyst was filtered off through a pad of celite and concentrated.
The residue was purified by column chromatography on silica gel
eluting with 30-40% ethyl acetate in hexane to provide
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-1-yl)-4-(4-hydro-
xyphenyl)pyrrolidine-2,5-dione 0.07 g as a light yellow solid. Mp
105-107.degree. C.; .sup.1H NMR (acetone-d.sub.6) 400 MHz .delta.:
10.27 (s, 1H), 8.34 (s, 1H), 7.17-7.21 (m, 4H), 6.80-6.91 (m, 4H),
4.39 (d, J=7.0 Hz, 1H), 4.22 (d, J=7.0 Hz, 1H), 4.13 (d, J=5.6 Hz,
2H), 2.92 (d, J=5.2 Hz, 2H), 2.15-2.19 (m, 2H).
Example 73
Preparation of
7-[4-(1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl]-3,4-dihydro-1H-
-[1,4]diazepino[6,7,1-hi]indole-2-carboxylic acid benzyl ester
Step 1
##STR00028##
[0310] To a solution of 7-formyl indole (2.4 g, 16.6 mmol) in
1,2-dichloroethane (60 mL) was added aminoethanol (1.2 mL, 19.8
mmol) followed by glacial acetic acid (2.0 mL) and sodium
triacetoxyborohydride (3.5 g, 16.6 mmol). The reaction mixture was
allowed to stir at room temperature for 16 hours. The reaction
mixture was quenched by addition of water (10 mL) and 1.0 M sodium
hydroxide (10 mL). The organic layer was then separated and the
aqueous layer extracted with 1,2-dichloroethane (40 mL). The
combined organic extracts were washed with saturated sodium
bicarbonate (2.times.30 mL), water (2.times.50 mL), dried over
anhydrous sodium sulfate and evaporated to dryness.
2-[(1H-indol-7-ylmethyl)-amino]-ethanol (4.4 g) was obtained as an
oil LCMS (M+H)=189.
Step 2
##STR00029##
[0312] To a solution of 2-[(1H-indol-7-ylmethyl)-amino]-ethanol
(4.4 g) in 1,2-dichloroethane (40 mL) was added triethylamine (4.85
mL, 34.6 mmol) followed by benzyl chloroformate (3.57 mL, 25.34
mmol). The mixture was allowed to stir at room temperature for 2
hours. The mixture was quenched by addition of water (20 mL), and
1.0 M sodium hydroxide (10 mL). The organic layer was separated and
the aqueous layer extracted with 1,2-dichloroethane (20 mL). The
combined organic extracts were washed with 1.0 M hydrochloric acid
(20 mL), water (20 mL), dried over anhydrous sodium sulfate and
evaporated to dryness. The residue was purified by silica gel
chromatography, eluting with 20% ethyl acetate in hexanes to 40%
ethyl acetate in hexanes to afford
(2-hydroxy-ethyl)-(1H-indol-7-ylmethyl)-carbamic acid benzyl ester
(2.79 g, 52% combined yield for two steps) as a colorless oil.
.sup.1H NMR (CDCl.sub.3) 400 MHz .delta.: 9.97 (br s, 1H), 7.75-6.9
(m, 8H), 6.54 (br s, 1H), 5.21 (s, 2H), 4.9-4.6 (m, 3H), 3.85-3.57
(m, 2H), 3.55-3.23 (m, 3H); LCMS M+H=325.
##STR00030##
[0313] To a solution of
(2-hydroxy-ethyl)-(1H-indol-7-ylmethyl)-carbamic acid benzyl ester
(2.79 g, 8.61 mmol) in dichloromethane (50 mL) was added
triethylamine (1.56 mL, 11.2 mmol). The mixture was cooled to
0.degree. C. and methanesulfonyl chloride (0.74 mL, 9.47 mmol)
added in a dropwise manner. The mixture was warmed to room
temperature and allowed to stir for 2 hours. The mixture was then
quenched with water (30 mL) and 1.0 M sodium hydroxide (10 mL). The
organic layer was separated and the aqueous layer extracted with
dichloromethane (20 mL). The combined extracts were washed with 1.0
M hydrochloric acid (20 mL), water (30 mL), dried over anhydrous
sodium sulfate and evaporated to dryness. Methanesulfonic acid
2-[benzyloxycarbonyl-(1H-indol-7-ylmethyl)-amino]-ethyl ester (3.46
g) was obtained as an oil
Step 4
##STR00031##
[0315] To a solution of methanesulfonic acid
2-[benzyloxycarbonyl-(1H-indol-7-ylmethyl)-amino]-ethyl ester (3.46
g, 8.61 mmol) in dimethylformamide (20 mL), which has been cooled
to 0.degree. C. was added sodium hydride (60%) in mineral oil. The
reaction mixture was allowed to stir at 0.degree. C. for 1 hour and
then quenched by the addition of water (40 mL). The aqueous layer
was extracted with ethyl acetate (4.times.20 mL). The combined
organic extracts were washed with water (3.times.30 mL), dried over
anhydrous sodium sulfate and evaporated to dryness. The residue was
purified by silica gel chromatography, eluting with dichloromethane
to afford
3,4-dihydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxylic acid
benzyl ester (1.95 g, 74% combined yield for two steps) as a
colorless oil. .sup.1H NMR (CDCL.sub.3) 400 MHz .delta.: 7.6-7.45
(m, 1H), 7.4-7.2 (m, 5H), 7.17-6.9 (m, 3H), 6.6-6.48 (m, 1H),
5.2-5.05 (m, 2H), 5.0-4.82 (m, 2H), 4.4-4.2 (m, 2H), 4.1-3.95 (m,
2H); LCMS (M+H)=307.
Step 5
##STR00032##
[0317] To a solution of
3,4-dihydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxylic acid
benzyl ester (557 mg, 1.8 mmol), in anhydrous tetrahydrofuran (1(1
ml) at 0.degree. C., was added oxalyl chloride (238 .mu.l, 2.7
mmol) followed by a further portion of oxalyl chloride (340 .mu.l,
3.85 mmol). The mixture was stirred at 0.degree. C. until all the
starting material has been consumed before being cooled to
-78.degree. C. Sodium methoxide in methanol (0.5M) (10 ml) was then
added slowly and the mixture allowed to warm to room temperature.
After 1 hour at room temperature the mixture was then diluted with
ethyl acetate (200 ml) and washed with water (300 ml). The organic
layer was dried over anhydrous sodium sulfate and evaporated to
dryness. The residue was purified by silica gel chromatography,
eluting with a ethyl acetate/hexanes (1:1) to afford
7-methoxyoxalyl-3,4-dihydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxyli-
c acid benzyl ester as a pale yellow solid (481 mg, 67%). .sup.1H
NMR (CDCl.sub.3) 400 MHz .delta.: 8.26-8.36 (m, 2H), 7.22-7.37 (m,
6H), 7.10 (dd, 1H, J=32.8 and 7.2 Hz), 5.11 (d, 2H, J=8.0 Hz), 4.94
(d, 2H, J=22.4 Hz), 4.41-4.48 (m, 2H), 4.01-4.05 (m, 2H), 3.93 (m,
3H).
Step 6
##STR00033##
[0319] To a solution of
7-methoxyoxalyl-3,4-dihydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxyli-
c acid benzyl ester (481 mg, 1.22 mmol) and indole-3-acetamide (234
mg, 1.34 mmol) in anhydrous tetrahydrofuran (14 ml) at 0.degree. C.
was added potassium t-butoxide (412 mg, 3.67 mmol). The mixture was
stirred at 0.degree. C. for 2 hours. Concentrated hydrochloric acid
(5 ml) was then added and the mixture stirred for 2 hours at room
temperature. The mixture was then diluted with ethyl acetate (300
ml), washed with water (500 ml), and the organic layer dried over
anhydrous sodium sulfate. The residue was purified by silica gel
chromatography, eluting with a ethyl acetate/hexanes (1:1) to
afford
7-[4-(1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl]-3,4-dihydro-1H-
-[1,4]diazepino[6,7,1-hi]indole-2-carboxylic acid benzyl ester as a
bright orange/red solid (1.2 g, 80%). .sup.1H NMR (DMSO-d.sub.6)
400 MHz .delta.: 11.66 (d, 1H, J=2.4 Hz), 10.94 (s, 1H), 7.69-7.75
(m, 2H), 7.19-7.38 (m, 6H), 6.98 (t, 1H, J=7.2 Hz), 6.73-6.89 (m,
3H), 6.60-6.66 (m, 2H), 4.90-5.08 (m, 2H), 4.50 (m, 2H), 3.95 (m,
2H).
Example 74
Preparation of
(.+-.)-Trans-7-[4-(1H-indol-3-yl)-2,5-dioxo-pyrrolidin-3-yl]-3,4-dihydro--
1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxylic acid benzyl
ester
##STR00034##
[0321] Magnesium turnings (195 mg, 8.0 mmol) were added to a
solution of
7-[4-(1H-indol-3-yl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-3-yl]-3,4-dihydro-1H-
-[1,4]diazepino[6,7,1-hi]indole-2-carboxylic acid benzyl ester (230
mg, 0.44 mmol) in anhydrous methanol (20 ml) and heated to reflux
under an atmosphere of nitrogen for 1.5 hours. After cooling to
room temperature the mixture was poured into ethyl acetate (200 ml)
and washed with 1 M hydrochloric acid (100 ml). The organic layer
was dried over anhydrous sodium sulfate and evaporated to dryness.
The residue was then purified by silica gel chromatography using
50-60% ethyl acetate in hexanes to yield
(.+-.)-trans-7-[4-(1H-indol-3-yl)-2,5-dioxo-pyrrolidin-3-yl]-3,4-di-
hydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxylic acid benzyl
ester as an off white solid (205 mg). .sup.1H NMR (DMSO-d.sub.6)
400 MHz .delta.: 1.56 (s, 1H), 11.03 (d, 1H, J=2 Hz), 7.21-7.43 (m,
10H), 7.09 (t, 1H, J=7.2 Hz), 6.92-7.00 (m, 2H), 6.82-6.89 (m, 3H),
5.04 (s, 2H), 4.87 (d, 2H, J=7.6 Hz), 4.54 (dd, 2H, J=7.6 and 28.8
Hz), 4.30 (m, 2H), 3.92 (m, 2H).
Example 75
Preparation of
(.+-.)-Trans-3-(1H-indol-3-yl)-4-(1,2,3,4-tetrahydro-[1,4]diazepino[6,7,1-
-hi]indol-7-yl)-pyrrolidine-2,5-dione
##STR00035##
[0323]
(.+-.)-Trans-7-[4-(1H-indol-3-yl)-2,5-dioxo-pyrrolidin-3-yl]-3,4-di-
hydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxylic acid benzyl
ester (161 mg, 0.31 mmol) and 10% palladium on carbon (100 mg) in
anhydrous methanol (15 ml) were stirred under 1 atmosphere of
hydrogen for 16 hours. The catalyst was then filtered through a bed
of Celite and the filtrate evaporated to dryness to yield
(.+-.)-trans-3-(1H-indol-3-yl)-4-(1,2,3,4-tetrahydro-[1,4]diazepino[6,7,1-
-hi]indol-7-yl)-pyrrolidine-2,5-dione as an off white solid (95
mg). .sup.1H NMR (DMSO-d.sub.6) 400 MHz .delta.: 11.04 (d, 1H,
J=1.6 Hz), 7.35-7.42 (m, 4H), 7.24 (dd, 1H, J=2.8 and 5.6 Hz), 7.09
(t, 1H, J=7.2 Hz), 6.89-6.98 (m, 3H), 4.52 (dd, 2H, J=7.2 and 24.8
Hz), 4.11 (s, 2H), 4.07-4.10 (m, 2H), 3.14-3.17 (m, 2H).
Example 76
[0324] Cell viability was determined by measuring the activity of
dehydrogenase enzymes in metabolically active cells using a
tetrazolium compound, MTS. The assay was performed as described in
Promega Technical Bulletin No. 169 (CellTiter 96 Aqueous
Non-Radioactive Cell Proliferation Assay). Thirteen human cancer
cell lines were assayed (see, e.g., Table 1). Cells were maintained
at 37.degree. C., 5% CO.sub.2. Adherent cells were maintained DMEM
media (4.5 g/L glucose) supplemented with 15% heat-inactivated FBS,
10 mM L-glutamine, and 10 mM Hepes pH 7.5. Suspension cells were
maintained in RPMI 1640 media, supplemented with 10%
heat-inactivated FBS and 10 mM Hepes pH 7.5. Briefly, cells were
seeded in 96-well plates as set forth in Table 1 and incubated for
16-24 hours. Candidate compounds were serially diluted in DMSO,
further diluted in cell culture media, and then added to cells
(final DMSO concentration of 0.33%). Cells were incubated in the
presence of candidate compound for 72 hours. MTS stock solution
(MTS 2 gm/L, PMS 46.6 mg/ml in PBS) was added to the cells (final
concentration MTS 2 gm/L and PMS 7.67 mg/L) and incubated for 4
hours. SDS was added to a final concentration of 1.4% and
absorbance at 490 nM was measured within two hours using a
platereader. The IC.sub.50 was defined as the concentration of
compound that results in a 50% reduction in the number of viable
cells as compared to control wells treated with DMSO only (0.33%)
and was calculated using non-linear regression analysis. IC.sub.50
values were given in Table 2 for the compounds listed.
TABLE-US-00001 TABLE 1 Cell Line Cancer Type Cells/well NCI-H69
small cell lung 8500 NCI-H446 small cell lung 10000 NCI-H82 small
cell lung 8500 HCT-116 small cell lung 1200 sHT29 colon 2500
MDA-MB-231 breast 3500 A549 lung 400 DU-145 prostate 1000 K562
chronic myelogenous leukemia 1200 MCF7 breast 8000 PC-3 prostate
3000 SK-MEL-28 melanoma 1000 SKOV-3 ovarian 1800
TABLE-US-00002 TABLE 2 Example DU- NCI- HCT- Compound Number A549
145 H69 116 HT29 K562 (.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo 2 2.89
[3,2,1-ij] quinolin-1-yl)-4(1H- indol-3-yl) pyrrolidine-2,5-dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 2, 3 10.7 9.61 1.82 4.55 0.424
pyrrolo [3,2,1-ij] quinolin-1-yl)- 4(1H-indol-3-yl)
pyrrolidine-2,5- dione 3(R),4(S)-3-(5,6-Dihydro-4H- 4 1.85 5.5
pyrrolo [3,2,1-ij] quinolin-1-yl)- 4(1H-indol-3-yl)
pyrrolidine-2,5- dione (faster eluting peak in supercritical
chromatography) 3(S),4(R)-3-(5,6-Dihydro-4H- 4 2.07 9.71 pyrrolo
[3,2,1-ij] quinolin-1-yl)- 4(1H-indol-3-yl) pyrrolidine-2,5- dione
(slower eluting peak in supercritical chromatography)
(-)-Trans-3-(5,6-dihydro-4H- 5 5.8 4 1.04 0.812 1.95 0.294 pyrrolo
[3,2,1-ij] quinolin-1-yl)- 4(1H-indol-3-yl) pyrrolidine-2,5- dione
(faster eluting peak in supercritical chromatography)
(+)-Trans-3-(5,6-dihydro-4H- 5 27.9 10.5 14.4 30.13 4.52 pyrrolo
[3,2,1-ij] quinolin-1-yl)- 4(1H-indol-3-yl) pyrrolidine-2,5- dione
(slower eluting peak in supercritical chromatography)
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 6 62.7 ij] quinolin-1-yl)-4(2-
trifluoromethyl-phenyl)-pyrrole-2, 5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 7 >100 ij]
quinolin-1-yl)-4-thiophen-2-yl- pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 9 >100 ij]
quinolin-1-yl)-4-pyridin-2-yl- pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 10 >100 ij]
quinolin-1-yl)-4(4-methoxy- phenyl)-pyrrole-2,5-dione
3-Benzo[1,3]dioxol-5-yl-4-(5,6- 11 >100 dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 12 66.5 ij]
quinolin-1-yl)-4-phenyl-pyrrole- 2,5-dione
3-Benzo[b]thiophen-2-yl-4-(5,6- 13 >100 dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 14 71.6 ij]
quinolin-1-yl)-4-(3-phenoxy- phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 15 75.8 ij]
quinolin-1-yl)-4-(3-chloro- phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 16 40.9 ij]
quinolin-1-yl)-4-(2-chloro- phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 17 >100 ij]
quinolin-1-yl)-4-(2,5- dimethoxy-phenyl)-pyrrole-2,5- dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 18 >100 ij]
quinolin-1-yl)-4-(2-chloro-4- fluoro-phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 19 41 >100 26.2 46.8 14 ij]
quinolin-1-yl)-4-naphthalene-1- yl-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 20 38.8 31.1 4.96 >100 2.47
ij] quinolin-1-yl)-4-(2,6-dichloro- phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 21 28.6 21.8 5.44 56.75 6.42 ij]
quinolin-1-yl)-4-(2-bromo- phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 22 >100 ij]
quinolin-1-yl)-4-indol-1-yl- pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 23 76.8 ij]
quinolin-1-yl)-4-pyridine-3-yl- pyrrole-2,5-dione
3-(4-Benzoyloxy-phenyl)-4-(5,6- 29 >100 dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-pyrrole-2,5-dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 37 37.4 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (5-(1-naphthyl)-1H-indol-3-yl)
pyrrolidine-2,5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 38 10.6
pyrrolo [3,2,1-ij] quinolin-1yl)-4-
(5-(4-methoxyphenyl)-1H-indol-3- yl) pyrrolidine-2,5-dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 39 10.5 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (5-(3-methylphenyl)-1H-indol-3- yl)
pyrrolidine-2,5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 40 10.5
pyrrolo [3,2,1-ij] quinolin-1yl)-4- (2-chloro-4-fluorophenyl)
pyrrolidine-2,5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 41 4.65
pyrrolo [3,2,1-ij] quinolin-1yl)-4- (2,6-dichlorophenyl)
pyrrolidine-2, 5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 42 44.1
pyrrolo [3,2,1-ij] quinolin-1yl)-4- (4-bromophenyl)
pyrrolidine-2,5- dione (.+-.)-Trans-3-(5,6-dihydro-4H- 43 48.7
pyrrolo [3,2,1-ij] quinolin-1yl)-4- (4-chlorophenyl)
pyrrolidine-2,5- dione (.+-.)-Trans-3-(5,6-dihydro-4H- 44 12.5
pyrrolo [3,2,1-ij] quinolin-1yl)-4- (4-trifluoromethoxyphenyl)
pyrrolidine-2,5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 45 27.6
pyrrolo [3,2,1-ij] quinolin-1yl)-4- (thiophen-3-yl)
pyrrolidine-2,5- dione (.+-.)-Trans-3-(5,6-dihydro-4H- 46 4.76
pyrrolo [3,2,1-ij] quinolin-1yl)-4- (2-fluorophenyl)
pyrrolidine-2,5- dione (.+-.)-Trans-3-(5,6-dihydro-4H- 47 4.45 18.3
pyrrolo [3,2,1-ij] quinolin-1yl)-4- (2-thiophen-2-yl)
pyrrolidine-2,5- dione (.+-.)-Trans-3-(5,6-dihydro-4H- 48 8.39
pyrrolo [3,2,1-ij] quinolin-1yl)-4- (2,4-dichlorophenyl)
pyrrolidine-2, 5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 49 14.6
pyrrolo [3,2,1-ij] quinolin-1yl)-4- phenyl-pyrrolidine-2,5-dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 50 5.41 8.51 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (2-chlorophenyl)-pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 51 37.7 66.75 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (N-methylindol-3-yl) pyrrolidine-2, 5-dione
(.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo 53 >100 >100 >100
>100 36.3 [3,2,1-ij] quinolin-1yl)-4-(2,5-
dimethoxyphenyl)-pyrrolidine-2,5- dione
(.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo 54 22.1 21.1 7.63 8.78 3.37
[3,2,1-ij] quinolin-1yl)-4-(2- chloro-4-fluoro-phenyl)-
pyrrolidine-2,5-dione (.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo 55 13.1
35.7 12.1 13.1 3.47 [3,2,1-ij] quinolin-1yl)-4-(3-
chlorophenyl)-pyrrolidine-2,5- dione Phosphoric acid dibenzyl ester
56 22.6 trans-3-(5,6-dihydro-4H- pyrrolo[3,2,1-ij]quinolin-1-yl)-4-
(1H-indol-3-yl)-2,5-dioxo- pyrrolidin-1-ylmethyl ester
(.+-.)-Phosphoric acid mono-[trans-3- 56 4.45
(5,6-dihydro-4H-pyrrolo[3,2,1- ij]quinolin-1-yl)-4-(1H-indol-3-yl)-
2,5-dioxo-pyrrolidin-1-ylmethyl] ester
(.+-.)-trans-2-Amino-propionic acid- 57 2.9
3-(5,6-dihydro-4H-pyrrolo[3,2,1-
ij]quinolin-1-yl)-4-(1H-indol-3-yl)- 2,5-dioxo-pyrrolidin-1-yl
methyl ester (.+-.)-trans-Amino-acetic acid-3-(5,6- 58 2.77
dihydro-4H-pyrrolo[3,2,1- ij]quinolin-1-yl)-4-(1H-indol-3-yl)-
2,5-dioxo-pyrrolidin-1-yl methyl ester
(.+-.)-trans-dimethylamino-acetic 59 3.74 acid-3-(5,6-dihydro-4H-
pyrrolo[3,2,1-ij]quinolin-1-yl)-4- (1H-indol-3-yl)-2,5-dioxo-
pyrrolidin-1-ylmethyl ester (.+-.)-trans-Isonicotinic acid-3-(5,6-
60 3.59 dihydro-4H-pyrrolo[3,2,1-
ij]quinolin-1-yl)-4-(1H-indol-3-yl)-
2,5-dioxo-pyrrolidin-1-ylmethyl ester
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo 61 78.9 >100 96.2 >100
18.7 [3,2,1-ij] quinolin-1-yl)-4(1- methylindol-3-yl)-1-methyl
pyrrolidine-2,5-dione 3-(5,6-Dihydro-4H-pyrrolo[3,2,1- 65 35
ij]quinolin-1-yl)-4-(3- methoxyphenyl)-1H-pyrrole-2,5- dione
3-[4-(Benzyloxy)phenyl]-4-(5,6- 66 >100
dihydro-4H-pyrrolo[3,2,1- ij]quinolin-1-yl)-1H-pyrrole-2,5- dione
3-(5,6-Dihydro-4H-pyrrolo[3,2,1- 67 53.1 ij]quinolin-1-yl)-4-(4-
fluorophenyl)-1H-pyrrole-2,5- dione (.+-.)-Trans-3-(5,6-dihydro-4H-
68 3.06 pyrrolo[3,2,1-ij]quinolin-1-yl)-4-
(3-methoxyphenyl)pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 69 55.4
pyrrolo[3,2,1-ij]quinolin-1-yl)-4-
(3-hydroxyphenyl)pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 70 26.2
pyrrolo[3,2,1-ij]quinolin-1-yl)-4- (4-fluorophenyl)pyrrolidine-2,5-
dione (.+-.)-Trans-3-[4- 71 48.7 (benzyloxy)phenyl]-4-(5,6-
dihydro-4H-pyrrolo[3,2,1- ij]quinolin-1-yl)pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 72 >100
pyrrolo[3,2,1-ij]quinolin-1-yl)-4-
(4-hydroxyphenyl)pyrrolidine-2,5- dione
7-[4-(1H-indol-3-yl)-2,5-dioxo- 73 >100
2,5-dihydro-1H-pyrrol-3-yl]-3,4- dihydro-1H-[1,4]diazepino[6,7,1-
hi]indole-2-carboxylic acid benzyl ester
(.+-.)-Trans-7-[4-(1H-indol-3-yl)- 74 90.7
2,5-dioxo-pyrrolidin-3-yl]-3,4- dihydro-1H-[1,4]diazepino[6,7,1-
hi]indole-2-carboxylic acid benzyl ester
(.+-.)-Trans-3-(1H-indol-3-yl)-4- 75 >100 (1,2,3,4-tetrahydro-
[1,4]diazepino[6,7,1-hi]indol-7-yl)- pyrrolidine-2,5-dione MDA- SK-
MB- NCI- NCI- MEL- SKOV- Compound MCF7 231 H446 H82 PC-3 28 3
(.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo 4.04 [3,2,1-ij]
quinolin-1-yl)-4(1H- indol-3-yl) pyrrolidine-2,5-dione
(.+-.)-Trans-3-(5,6-dihydro-4H- >100 3.34 4.95 13 0.939 pyrrolo
[3,2,1-ij] quinolin-1-yl)- 4(1H-indol-3-yl) pyrrolidine-2,5- dione
3(R),4(S)-3-(5,6-Dihydro-4H- 3.69 1.27 1.49 pyrrolo [3,2,1-ij]
quinolin-1-yl)- 4(1H-indol-3-yl) pyrrolidine-2,5- dione (faster
eluting peak in supercritical chromatography)
3(S),4(R)-3-(5,6-Dihydro-4H- 5.17 1.42 1.77 pyrrolo [3,2,1-ij]
quinolin-1-yl)- 4(1H-indol-3-yl) pyrrolidine-2,5- dione (slower
eluting peak in supercritical chromatography)
(-)-Trans-3-(5,6-dihydro-4H- >100 2.19 0.581 0.828 1.28 8.68
0.444 pyrrolo [3,2,1-ij] quinolin-1-yl)- 4(1H-indol-3-yl)
pyrrolidine-2,5- dione (faster eluting peak in supercritical
chromatography) (+)-Trans-3-(5,6-dihydro-4H- >100 26.83 8.55
9.83 18.7 25.5 4.45 pyrrolo [3,2,1-ij] quinolin-1-yl)-
4(1H-indol-3-yl) pyrrolidine-2,5- dione (slower eluting peak in
supercritical chromatography) 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-
61.2 ij] quinolin-1-yl)-4(2- trifluoromethyl-phenyl)-pyrrole-2,
5-dione 3-(5,6-Dihydro-4H-pyrrolo [3,2,1- >100 ij]
quinolin-1-yl)-4-thiophen-2-yl- pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- >100 ij]
quinolin-1-yl)-4-pyridin-2-yl- pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- >100 ij]
quinolin-1-yl)-4(4-methoxy- phenyl)-pyrrole-2,5-dione
3-Benzo[1,3]dioxol-5-yl-4-(5,6- >100 dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 81.6 ij]
quinolin-1-yl)-4-phenyl-pyrrole- 2,5-dione
3-Benzo[b]thiophen-2-yl-4-(5,6- 78.8 dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-pyrrole-2,5-dione 3-(5,6-Dihydro-4H-pyrrolo [3,2,1-
98.2 ij] quinolin-1-yl)-4-(3-phenoxy- phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 43.6 ij]
quinolin-1-yl)-4-(3-chloro- phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 21.5 ij]
quinolin-1-yl)-4-(2-chloro- phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- >100 ij]
quinolin-1-yl)-4-(2,5- dimethoxy-phenyl)-pyrrole-2,5- dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- >100 ij]
quinolin-1-yl)-4-(2-chloro-4- fluoro-phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- >100 70.5 96.2 39.6 36.2 ij]
quinolin-1-yl)-4-naphthalene-1- yl-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- >100 50.3 82.7 74 23.8 ij]
quinolin-1-yl)-4-(2,6-dichloro- phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- >100 30 22.2 >100 30.2 ij]
quinolin-1-yl)-4-(2-bromo- phenyl)-pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 59.4 ij]
quinolin-1-yl)-4-indol-1-yl- pyrrole-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo [3,2,1- 28.7 ij]
quinolin-1-yl)-4-pyridine-3-yl- pyrrole-2,5-dione
3-(4-Benzoyloxy-phenyl)-4-(5,6- >100 dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-pyrrole-2,5-dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 43.4 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (5-(1-naphthyl)-1H-indol-3-yl)
pyrrolidine-2,5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 13.1 pyrrolo
[3,2,1-ij] quinolin-1yl)-4- (5-(4-methoxyphenyl)-1H-indol-3- yl)
pyrrolidine-2,5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 11.2 pyrrolo
[3,2,1-ij] quinolin-1yl)-4- (5-(3-methylphenyl)-1H-indol-3- yl)
pyrrolidine-2,5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 10.7 pyrrolo
[3,2,1-ij] quinolin-1yl)-4- (2-chloro-4-fluorophenyl)
pyrrolidine-2,5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 7.63 pyrrolo
[3,2,1-ij] quinolin-1yl)-4- (2,6-dichlorophenyl) pyrrolidine-2,
5-dione (.+-.)-Trans-3-(5,6-dihydro-4H- 49.6 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (4-bromophenyl) pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 54.5 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (4-chlorophenyl) pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 14.1 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (4-trifluoromethoxyphenyl) pyrrolidine-2,5-dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 42.7 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (thiophen-3-yl) pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- >100 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (2-fluorophenyl) pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 19.87 9.82 7.25 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (2-thiophen-2-yl) pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 14.8 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (2,4-dichlorophenyl) pyrrolidine-2, 5-dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 12 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- phenyl-pyrrolidine-2,5-dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 4.56 4.22 4.65 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (2-chlorophenyl)-pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 67.3 65.7 37.7 pyrrolo [3,2,1-ij]
quinolin-1yl)-4- (N-methylindol-3-yl) pyrrolidine-2, 5-dione
(.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo >100 >100 >100
>100 57.4 [3,2,1-ij] quinolin-1yl)-4-(2,5-
dimethoxyphenyl)-pyrrolidine-2,5- dione
(.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo >100 9.3 11.5 25.2 6.88
[3,2,1-ij] quinolin-1yl)-4-(2- chloro-4-fluoro-phenyl)-
pyrrolidine-2,5-dione (.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo >100
15.54 12.9 >100 11.6 [3,2,1-ij] quinolin-1yl)-4-(3-
chlorophenyl)-pyrrolidine-2,5- dione Phosphoric acid dibenzyl ester
>100 trans-3-(5,6-dihydro-4H- pyrrolo[3,2,1-ij]quinolin-1-yl)-4-
(1H-indol-3-yl)-2,5-dioxo- pyrrolidin-1-ylmethyl ester
(.+-.)-Phosphoric acid mono-[trans-3- 5.89
(5,6-dihydro-4H-pyrrolo[3,2,1- ij]quinolin-1-yl)-4-(1H-indol-3-yl)-
2,5-dioxo-pyrrolidin-1-ylmethyl] ester
(.+-.)-trans-2-Amino-propionic acid- 3.14
3-(5,6-dihydro-4H-pyrrolo[3,2,1-
ij]quinolin-1-yl)-4-(1H-indol-3-yl)- 2,5-dioxo-pyrrolidin-1-yl
methyl ester (.+-.)-trans-Amino-acetic acid-3-(5,6- 3.45
dihydro-4H-pyrrolo[3,2,1- ij]quinolin-1-yl)-4-(1H-indol-3-yl)-
2,5-dioxo-pyrrolidin-1-yl methyl ester
(.+-.)-trans-dimethylamino-acetic 4.47 acid-3-(5,6-dihydro-4H-
pyrrolo[3,2,1-ij]quinolin-1-yl)-4- (1H-indol-3-yl)-2,5-dioxo-
pyrrolidin-1-ylmethyl ester (.+-.)-trans-Isonicotinic acid-3-(5,6-
3.55 dihydro-4H-pyrrolo[3,2,1- ij]quinolin-1-yl)-4-(1H-indol-3-yl)-
2,5-dioxo-pyrrolidin-1-ylmethyl ester
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo >100 >100 >100
>100 37.6 [3,2,1-ij] quinolin-1-yl)-4(1-
methylindol-3-yl)-1-methyl pyrrolidine-2,5-dione
3-(5,6-Dihydro-4H-pyrrolo[3,2,1- 60.8 ij]quinolin-1-yl)-4-(3-
methoxyphenyl)-1H-pyrrole-2,5- dione
3-[4-(Benzyloxy)phenyl]-4-(5,6- >100 dihydro-4H-pyrrolo[3,2,1-
ij]quinolin-1-yl)-1H-pyrrole-2,5- dione
3-(5,6-Dihydro-4H-pyrrolo[3,2,1- 57 ij]quinolin-1-yl)-4-(4-
fluorophenyl)-1H-pyrrole-2,5- dione (.+-.)-Trans-3-(5,6-dihydro-4H-
14.7 pyrrolo[3,2,1-ij]quinolin-1-yl)-4-
(3-methoxyphenyl)pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 65.9
pyrrolo[3,2,1-ij]quinolin-1-yl)-4-
(3-hydroxyphenyl)pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- 49.2
pyrrolo[3,2,1-ij]quinolin-1-yl)-4- (4-fluorophenyl)pyrrolidine-2,5-
dione (.+-.)-Trans-3-[4- 97.5 (benzyloxy)phenyl]-4-(5,6-
dihydro-4H-pyrrolo[3,2,1- ij]quinolin-1-yl)pyrrolidine-2,5- dione
(.+-.)-Trans-3-(5,6-dihydro-4H- >100
pyrrolo[3,2,1-ij]quinolin-1-yl)-4-
(4-hydroxyphenyl)pyrrolidine-2,5- dione
7-[4-(1H-indol-3-yl)-2,5-dioxo- >100
2,5-dihydro-1H-pyrrol-3-yl]-3,4- dihydro-1H-[1,4]diazepino[6,7,1-
hi]indole-2-carboxylic acid benzyl ester
(.+-.)-Trans-7-[4-(1H-indol-3-yl)- >100
2,5-dioxo-pyrrolidin-3-yl]-3,4- dihydro-1H-[1,4]diazepino[6,7,1-
hi]indole-2-carboxylic acid benzyl ester
(.+-.)-Trans-3-(1H-indol-3-yl)-4- >100 (1,2,3,4-tetrahydro-
[1,4]diazepino[6,7,1-hi]indol-7-yl)- pyrrolidine-2,5-dione
Example 77
[0325] Exponentially growing MDA-MB-231 cells or Paca-2 cells were
seeded at 1,000 cells per well in six-well plates and allowed to
attach for 24 hours. MDA-MB-231 and Paca-2 cells were cultured in
DMEM supplemented with 10% (v/v) fetal calf serum (FCS) and 5 ml
Penicillin/Streptomycin at 37.degree. C. in 5% CO.sub.2. MDA-MB-231
and Paca-2 were established estrogen receptor negative human breast
cancer and pancreatic carcinoma cell lines, respectively.
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij]quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij]quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
were each dissolved at a concentration of 10 mM in DMSO, and
separately added to cells at a concentration of 0.1, 0.25, 0.5, 1
or 2 .mu.M. Control plates received DMSO alone, at the same
percentage of total culture volume as that administered in
conjunction with the highest concentration of drug. Cell cultures
were observed daily for 10-15 days, then fixed and stained with
modified Wright-Giemsa stain (Sigma). Treatment with
(.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione results in
cell death of MDA-MB-231 cells or Paca-2 cells. See, e.g., FIG. 2.
The IC.sub.50 for (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione was found to
be 0.5 .mu.M. The IC.sub.50 for
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione was found to
be 0.5 .mu.M.
Example 78
[0326] MDA-MB-231 cells (ATCC# HTB-26), grown in DMEM plus 15% heat
inactivated fetal bovine serum plus 10 mM HEPES pH 7.5, were plated
in 60 mm plates (2.times.10.sup.5 cells per plate). After two days
candidate compounds in DMSO at various concentrations were diluted
in media and added to individual plates such that the final DMSO
concentration in the cell culture media was 0.1%. After two days
incubation the culture was trypsinized, cells were washed with
media, counted using a hemocytometer and 500 cells, including cell
bodies, were plated in 100 mM plates in media. Two weeks later the
media was removed and the cell colonies were fixed with methanol
for 10 minutes, stained with 1% crystal violet 10 minutes, washed
with water and airdried. Cell colonies were visually counted when
there were greater then 50 cells present per colony. Plating
efficiency was defined as the average number of colonies formed
divided by 500. The surviving fraction was defined as the plating
efficiency of a candidate compound divided by the plating
efficiency of DMSO multiplied by 100. For candidate compound
titrations, the IC.sub.50 concentration was determined by fitting
the equation y=Ac.sup.Bx to the data points and extrapolating the
concentration where surviving fraction equaled 50. Treatment with
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione results in
cell death of MDA-MB-231 cells. See. e.g., FIG. 3. The IC.sub.50
for (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione was found to
be 0.62 .mu.M. The IC.sub.50 for
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione was found to
be 4.1 .mu.M.
Example 79
[0327] Recombinant Protein Kinase C (Calbiochem) (100 ng) was
incubated with (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione at 0.05,
0.5, or 10 .mu.M for 15 minutes at room temperature. Subsequently,
a radioactive labeling mix in kinase buffer (20 mM Tris-HCl pH 7.5,
10 mM MgCl.sub.2) containing 20 .mu.M ATP, 0.2 .mu.Ci/.mu.l
.gamma..sup.32P-ATP, 0.2 .mu.g/.mu.l Histone H1 (Upstate) was added
to each sample. The kinase reaction was carried out for 5 minutes
at room temperature. Reaction products were analyzed by 12%
SDS-PAGE and autoradiography.
[0328] Treatment of recombinant Protein Kinase C for 15 minutes at
room temperature with (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione at the
tested concentrations did not reduce kinase activity in comparison
to treatment with carrier alone. See. e.g., FIG. 4.
Example 80
[0329] MDA-MB-231 cells were serum-deprived overnight (16 hours) in
the absence or in the presence of the indicated concentrations of
the separate enantiomers (.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
and (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione. Cells were
treated with 100 ng/ml recombinant human Hepatocyte Growth
Factor/Scatter Factor (HGF/SF) (R&D Systems #294-HG) for 10
minutes. Whole cell extracts were prepared in lysis buffer (20 mM
Trwas-HCl pH 7.5, 150 mM NaCl, 1 mM Na.sub.2EDTA, 1 mM EGTA, 1%
Triton X-100, 2.5 mM sodium pyrophosphate, 1 mM
beta-glycerophosphate, 1 mM Na.sub.3VO.sub.4, 1 .mu.g/ml leupeptin,
1 mM phenylmethylsulfonyl fluoride) and sonicated. Protein
concentration was measured by Bradford assay using the BioRad
reagent (BioRad, Hercules, Calif.), according to the manufacturer's
directions. Samples (50 .mu.g of protein) were resolved by 8%
SDS-PAGE under reducing conditions, and transferred onto a PVDF
membrane (BioRad). The membrane was incubated 1 hour in TBS-T (50
mM Trwas-HCl pH 7.6, 200 mM NaCl, 0.05% Tween 20) with 5% milk.
Proteins were detected by incubation overnight at 4.degree. C. in
TBS-T with 5% milk and either a polyclonal antibody against
phosphorylated c-Met (#3121), phosphorylated Erk1/2 (#9101), total
Erk1/2 protein (#9102) (Cell Signaling Technology); or a monoclonal
antibody against .beta.-actin (A-5441) (Sigma), which was used as a
control for total protein loading. After extensive washing in
TBS-T, a horseradish peroxidase-conjugated anti-rabbit IgG (1:5000)
or anti-mouse IgG (1:2000) (Amersham Biosciences) was added for 1
hour, and specific protein bands visualized using an enhanced
chemiluminescence detection system (Amersham Biosciences),
according to the manufacturer's instructions. See. e.g., FIG.
5.
[0330] Treatment with (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
inhibits both basal and HGF-induced autophosphorylation of c-Met at
a concentration of at least 500 nM.
(+)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione has a
partial inhibitory effect on c-Met phosphorylation at greater
concentrations (10 to 20 .mu.M). Additionally,
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione decreases
the phosphorylation of ERK1/2; a well-known downstream target in
the signaling of the c-Met tyrosine kinase receptor. See, e.g.,
FIG. 5.
Example 81
[0331] Athymic female nude mice (CRL:NU/NU-nuBR) were injected
subcutaneously in the flank with MDA-M3-231 human breast cancer
cells (8.times.10.sup.6 cells/mouse). Prior to injection,
MDA-MB-231 cells were cultured in DMEM (Invitrogen #11965-092)
supplemented with 10% (v/v) heat inactivated fetal bovine serum
(FBS), 1% Hepes buffer solution (Invitrogen #15630-080) and 1%
Penicillin/Streptomycin (Invitrogen #15140-122) at 37.degree. C. in
5% CO.sub.2. Tumors were allowed to grow to approximately 50
mm.sup.3 in size. Animals were randomized into three groups of five
animals per group. At day ten post injection, mice with established
tumors were treated intraperitoneally with
(+)-cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione (160 mg/kg),
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione (160 mg/kg)
administered in iodinated poppy seed oil (lipiodol) as the vehicle
at 10 mg/ml, or vehicle control. Drug or vehicle was administered
every three clays for a total of ten doses (q3dx10). Tumor size was
evaluated periodically during the study. For each subject, tumor
volume was calculated using the formula (L.times.W.sup.2)/2 where L
and W were the length and width of the tumor, respectively. The
arithmetic mean tumor volume was calculated for each treatment
group+/-standard error of the mean (SEM). The means for each group
were normalized with respect to the mean starting tumor volume in
order to determine fold increase in tumor volume. See. e.g., FIG.
6.
[0332] Treatment with (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-drone at 160 mg/kg
reduced the normalized mean tumor volume of human breast cancer
xenograft by 80.5% (p=0.0037) and 43.5% (p=0.133), respectively,
compared to vehicle treated control. See, e g., FIG. 6.
Example 82
[0333] Athymic female nude mice (CRL:NU/NU-nuBR) were injected
subcutaneously in the flank with MDA-MB-231 human breast cancer
cells (8.times.10.sup.6 cells/mouse). Prior to injection,
MDA-MB-231 cells were cultured in DMEM (Invitrogen #11965-092)
supplemented with 10% (v/v) heat inactivated fetal bovine serum
(FBS), 1% Hepes buffer solution (Invitrogen #15630-080) and 1%
Penicillin/Streptomycin (Invitrogen #15140-122) at 37.degree. C. in
5% CO.sub.2. Tumors were allowed to grow to approximately 50 mm in
size. Animals were randomized into three groups of eight animals
per group. At day four post injection, mice with established tumors
were treated intraperitoneally with
(.+-.)-Cis-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione (80 mg/kg),
(.+-.)-Trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione (80 mg/kg)
administered in DMA/PEG 400 (1:4 v/v) as the vehicle at 50 mg/ml,
or vehicle control intraperitoneally. Drug or vehicle was
administered once per day for five consecutive days followed by two
consecutive days of no treatment (one cycle of dose
administration). A total of four cycles of treatment were
administered. Tumor size was evaluated periodically during the
study. For each subject, tumor volume was calculated using the
formula (L.times.W.sup.2)/2 where L and W were the length and width
of the tumor, respectively. The arithmetic mean tumor volume was
calculated for each treatment group+/-standard error of the mean
(SEM). Each data point in FIG. 6 represents the arithmetic
mean+/-standard error of the mean (SEM) of eight tumors.
[0334] Treatment with (.+-.)-cis-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or
(.+-.)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione at 80 mg/kg
reduced the mean tumor volume of human breast cancer xenograft by
73.4% (p=0.04) and 69.4% (p=0.075), respectively, compared to
vehicle treated control. See. e.g., FIG. 7.
Example 83
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione Induces Cell
Death of a Wide Range of Tumor Cells, Which Correlates with its
Ability of Inhibiting c-Met Phosphorylation
TABLE-US-00003 [0335] (-)-trans IC.sub.50 Expression Constitutive
(.mu.M) levels activity c-Met Cells Tissue of c-Met of c-Met
Inhibition CFA H661 Lung Null no na 5 (NSCLC) H446 Lung Null no na
4 (SCLC) MCF-7 Breast Low yes 0.1 0.4 MDA-MB- Breast Medium no 0.1
0.3 231 DLD-1 Colon Medium yes 3 0.7 HT-29 Colon High no 0.3 0.2
PACA2 Pancreas High yes 0.1 0.3 PANC-1 Pancreas High no 0.5 0.5
H441 Lung Very high no 0.1 0.2 (NSCLC) CFA = Colony Formation Assay
na = Not Applicable
[0336] To determine c-met inhibition, exponentially growing MCF-7
and MDA-MB-231 human breast cancer cells, DLD1 and HT29 human colon
cancer cells, PACA2 and PANC-1 human pancreatic cancer cells and
NCI-H441 human non-small cell lung cancer cells (H441) were serum
starved in 0.5% FBS and treated with increasing amounts of
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione for 24 hours
as indicated. Cells were then left unstimulated or stimulated with
100 ng/mL HGF for 10 minutes and whole cell extracts were prepared.
Whole cell lysates (50 .mu.g) were resolved by SDS-PAGE,
transferred onto a PVDF membrane, and an enhanced chemiluminescence
assay system (ECL) was used to determine the phosphorylation status
of c-Met. A polyclonal antibody against phospho (Y1349) c-Met
(phospho-c-Met) was obtained from Biosource International and a
monoclonal antibody against .beta.-actin was purchased from
Invitrogen. Percent inhibition was determined by densitometry using
the Scion Image
[0337] For colony formation assays (CFA), exponentially growing
cells were seeded at 2,000 cells per well in 6-well plates and
allowed to attach for 24 hours. Increasing concentrations of
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione (from 0.01
to 30 .mu.M) were then added to the media for another 24 hours.
After 24 hours exposure, the drug was removed and fresh media was
added for the next 14-21 days, allowing for colony formation. Cells
were fixed and stained with GEMSA (Gibco BRL). Colonies of greater
than 50 cells were scored as survivors and percentage of cell
survival was plotted to determine the IC.sub.50.
[0338] (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-done was found to
potently inhibit growth and induce apoptosis across a wide range of
human tumor cell lines. The IC.sub.50 of
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione for c-Met
expressing solid tumor types is in the 0.1-0.7 .mu.M range
following exposure to (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
for 24 hours, as determined by colony formation assay (CFA). More
notably cells lacking c-Met such as NCI-H661 (human non-small cells
lung cancer cells) and NCI-H446 (human small lung cancer cells)
yielded IC.sub.50s approximately 10-fold higher for
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione, strongly
indicating a correlation between the presence of c-Met and the
cytotoxic sensitivity of the cells towards
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione. It should
be noted that the IC.sub.50 of (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
on c-Met inhibition and cellular cytotoxicity after 24 hours
treatment are highly comparable in c-Met expressing cancer
cells.
Example 84
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione Induces
Apoptosis in Cancer Cells
[0339] A549 human lung cancer cells in a 96-well plate (Costar
3603, 5,000/well) were treated with either A) DMSO as a control; B)
1.2 .mu.M (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione for 38 hours
before addition of 1:200 fluorescent Annexin V (green) and 1:500
Propidium iodide (magenta, final concentration of 1 .mu.g/mL). The
labeling procedure was allowed to process at 37.degree. C. for 20
minutes followed by image acquisition and analysis using an IC100
Image Cytometer (Beckman Coulter, Inc) with 10.times.
amplification.
[0340] To determine whether (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
works primarily through a cytostatic or apoptotic mechanism, cancer
cells exposed to (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione were stained
with fluorescently labeled Annexin V (green fluorescence) and
propidium iodide (bright magenta fluorescence). Annexin V is a
well-validated reagent that specifically binds with high affinity
to externalized membrane phosphatidylserine, an early marker of the
onset of apoptosis, while propidium iodide is a marker for dead
cells. Incubation of human lung cancer cells (A549) with 1.2 .mu.M
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione for 38 hours
induced cells to undergo apoptosis as evidenced by strong Annexin V
staining. A small percentage of cells (.about.10-20%) co-stain with
both Annexin V and propidium iodide, indicating that a
sub-population of (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij]quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
treated cells were already dead within 38 hours. These data are
consistent with (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione inducing
cell death largely through activation of apoptotic mechanisms. (See
FIG. 8)
Example 85
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione Inhibits
Metastatic Cancer Cell Invasion
[0341] MDA-MB-231 cells were pretreated with indicated
concentrations of (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione for 24
hours. 300 .mu.l of each cell suspension (at a concentration
0.5.times.10.sup.6 cells/mL in serum free medium) was placed in
individual inserts and incubated for 24 hours at 37.degree. C. The
bottom wells housing the inserts contained 500 .mu.l of 10% FBS
medium. At 24 hours the media from the inserts was aspirated and
cells that failed to invade were gently removed from the interior
of the inserts with a cotton tipped swab. Each insert was then
transferred to a clean well containing cell stain solution and
incubated for 10 minutes at room temperature. The bottom of the
insert was destained by incubating in extraction solution and OD
was measured at 560 nM. (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
inhibited the migration across interstices in confluent cultures of
MDA-MB-231 cancer cells. The data represent the mean of two
independent experiments. (See FIG. 9)
[0342] The morbidity and mortality resulting from most cancers is
the result of local invasion and metastasis from the primary tumors
to other tissues. This process mostly depends on the motility and
growth of tumor cells. Activation of c-Met by HGF induces a variety
of cellular responses including motility, invasion, wound healing
and tissue regeneration. It has been established that aberrant
activation of c-Met plays a critical role in the development and
progression of primary tumors and secondary metastases. HGF has the
ability to dissociate epithelial sheets, to stimulate cell motility
and invasion through extracellular matrix substrates and HGF
production correlates with tumor metastasis in vivo.
[0343] As shown above, (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
inhibited the invasive phenotype of MDA-MB-231 breast cancer cells
with an estimated IC.sub.50 of approximately 500 nM. Similar
results were seen with brain and lung cancer cells (data not
shown).
Example 86
Breast Cancer Xenograft Model
[0344] (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione shows
efficacy in a human breast cancer xenograft. MDA-MB-231 human
breast cancer cells were inoculated subcutaneously into female
athymic nude mice (8.0.times.10.sup.6 cells/mouse) and allowed to
form palpable tumors. Once the tumors reached approximately 60
mm.sup.3, the animals were treated orally with
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione at 200 mg/kg
or vehicle control daily (5 consecutive days, followed by a 2 day
dosing holiday). (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione was
formulated in PEG 400:20% Vitamin E TPGS (60:40). The animals
received a total of 20 doses of (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
or vehicle control. Tumors were measured throughout treatment and
the post-treatment observation period. Each point represents the
mean.+-.SEM of ten tumors. (See FIG. 10)
[0345] Treatment with (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
as monotherapy was effective at slowing tumor growth. Tumor growth
inhibition of (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione was
calculated to be 79% and was statistically significant (p=0.009).
There was no significant change in body weight due to oral
administration of the vehicle or
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione at 200
mg/kg.
Example 87
Colon Cancer Xenograft Model
[0346] In this human colon cancer xenograft model HT29 human colon
cancer cells were inoculated subcutaneously into female athymic
nude mice (5.times.10.sup.6 cells/mouse) and allowed to form
palpable tumors. Once the tumors reached approximately 60 mm.sup.3,
the animals were treated orally with either
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione at 200 mg/kg
or 300 mg/kg, or vehicle control daily (5 consecutive days,
followed by a 2 day dosing holiday).
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione was
formulated in PEG 400:20% Vitamin E TPGS (60:40). The animals
received a total of 20 treatments of either
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione or vehicle
control. Tumors were measured throughout treatment and the
post-treatment observation period. Each point represents the
mean.+-.SEM of ten tumors. (See FIG. 11)
[0347] In this highly aggressive colon xenograft model, animals
dosed with either 200 mg/kg or 300 mg/kg as a monotherapy showed
significant tumor growth inhibition, with 300 mg/kg being more
efficacious than 200 mg/kg. (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
dosed at 200 mg/kg showed an optimal tumor growth inhibition of 39%
(p=0.006) while 300 mg/kg showed an optimal tumor growth inhibition
of 55% (p=0.00001). There was no significant change in body weight
due to oral administration of either vehicle control or
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione at 200 mg/kg
or 300 mg/kg.
Example 88
Pancreatic Cancer Xenograft Model
[0348] (-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione shows
efficacy in a human pancreatic cancer xenograft model. PACA-2 human
pancreatic cancer cells were inoculated subcutaneously into female
athymic nude mice (5.times.10.sup.6 cells/mouse) and allowed to
form palpable tumors. Once the tumors reached approximately 60
mm.sup.3, the animals were treated orally with
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione at 200 mg/kg
or 300 mg/kg or vehicle control daily (5 consecutive days, followed
by a 2 day dosing holiday). (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
was formulated in PEG 400:20% Vitamin E TPGS (60:40). The animals
received a total of 20 doses of (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
or vehicle control. Tumors were measured throughout treatment and
the post-treatment observation period. Each point represents the
mean.+-.SEM of ten tumors. (See FIG. 12)
[0349] Treatment with (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
as a monotherapy at either 200 mg/kg or 300 mg/kg showed
significant tumor growth inhibition, with 200 mg/kg and 300 mg/kg
being equally effective. (-)-trans-3-(5,6-dihydro-4H-pyrrolo
[3,2,1-ij] quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione
dosed at 200 mg/kg showed an optimal tumor growth inhibition of 58%
(p=0.036) while 300 mg/kg showed an optimal tumor growth inhibition
of 60% (p=0.018). There was no significant change in body weight
due to oral administration of either vehicle control or
(-)-trans-3-(5,6-dihydro-4H-pyrrolo [3,2,1-ij]
quinolin-1-yl)-4(1H-indol-3-yl) pyrrolidine-2, 5-dione at 200 mg/kg
and 300 mg/kg.
[0350] Other embodiments were within the following claims. While
several embodiments have been shown and described, various
modifications may be made without departing from the spirit and
scope of the present invention.
* * * * *
References