U.S. patent application number 13/628446 was filed with the patent office on 2013-11-14 for methods of inhibiting the catalytic activity of a protein kinase and of treating a protein kinase related disorder.
This patent application is currently assigned to Shanghai Hengrui Pharmaceutical Co., Ltd.. The applicant listed for this patent is JIANGSU HANSOH PHARMACEUTICAL CO., LTD., SHANGHAI HENGRUI PHARMACEUTICAL CO., LTD.. Invention is credited to Pingyan Bie, Minggang Ju, Yali Li, Guangtao Qian, Yidong Su, Peng Cho Tang, Jialiang Yang, Lei Zhang, Fuqiang Zhao, Ying Zhou.
Application Number | 20130303518 13/628446 |
Document ID | / |
Family ID | 40001668 |
Filed Date | 2013-11-14 |
United States Patent
Application |
20130303518 |
Kind Code |
A1 |
Tang; Peng Cho ; et
al. |
November 14, 2013 |
Methods of Inhibiting the Catalytic Activity of a Protein Kinase
and of Treating a Protein Kinase Related Disorder
Abstract
The invention provides a method of inhibiting the catalytic
activity of a protein kinase comprising contacting said protein
kinase with a compound of formula (I), or a pharmaceutically
acceptable salt thereof, or a pharmaceutical composition containing
the same, and also provides a method of treating a protein kinase
related disorder comprising a step of administering to a patient in
need thereof a therapeutically effective amount of the compound of
formula (I) or a pharmaceutically acceptable salt thereof, or a
pharmaceutical composition containing the same, wherein each
substituent in formula (I) is same as defined in the description.
##STR00001##
Inventors: |
Tang; Peng Cho; (Shanghai,
CN) ; Su; Yidong; (Shanghai, CN) ; Li;
Yali; (Shanghai, CN) ; Zhang; Lei; (Shanghai,
CN) ; Zhao; Fuqiang; (Shanghai, CN) ; Yang;
Jialiang; (Shanghai, CN) ; Zhou; Ying;
(Shanghai, CN) ; Bie; Pingyan; (Shanghai, CN)
; Qian; Guangtao; (Shanghai, CN) ; Ju;
Minggang; (Shanghai, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
JIANGSU HANSOH PHARMACEUTICAL CO., LTD.
SHANGHAI HENGRUI PHARMACEUTICAL CO., LTD. |
Lianyungang City
Shanghai |
|
CN
CN |
|
|
Assignee: |
Shanghai Hengrui Pharmaceutical
Co., Ltd.
Shanghai
CN
Jiangsu Hansoh Pharmaceutical Co., Ltd.
Lianyungang City
CN
|
Family ID: |
40001668 |
Appl. No.: |
13/628446 |
Filed: |
September 27, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
12451466 |
Dec 3, 2009 |
8329682 |
|
|
PCT/CN2008/001352 |
May 14, 2008 |
|
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13628446 |
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Current U.S.
Class: |
514/212.06 ;
435/184; 514/414 |
Current CPC
Class: |
A61P 3/10 20180101; A61P
17/06 20180101; A61P 35/02 20180101; A61P 35/00 20180101; A61P
37/06 20180101; A61P 43/00 20180101; A61P 29/00 20180101; A61P
41/00 20180101; C07D 487/04 20130101; A61P 19/02 20180101; A61P
9/00 20180101; A61P 35/04 20180101; A61P 37/02 20180101 |
Class at
Publication: |
514/212.06 ;
435/184; 514/414 |
International
Class: |
A61K 31/55 20060101
A61K031/55; A61K 31/407 20060101 A61K031/407; A61P 35/02 20060101
A61P035/02; A61P 35/00 20060101 A61P035/00; A61P 17/06 20060101
A61P017/06; A61P 19/02 20060101 A61P019/02; A61P 9/00 20060101
A61P009/00; A61P 29/00 20060101 A61P029/00; C12N 9/99 20060101
C12N009/99; A61P 3/10 20060101 A61P003/10 |
Foreign Application Data
Date |
Code |
Application Number |
May 14, 2007 |
CN |
200710107463.8 |
Apr 11, 2008 |
CN |
200810087564.8 |
Claims
1. A method of inhibiting the catalytic activity of a protein
kinase comprising contacting said protein kinase with a compound of
formula (I): ##STR00203## or a pharmaceutically acceptable salt
thereof, or a pharmaceutical composition containing the same,
wherein the inhibiting of catalytic activity occurs in non-colon
cancer treatment activity; X being selected from the group
consisting of carbon and nitrogen; R.sub.1 and R.sub.2 each being
independently selected from the group consisting of hydrogen and
alkyl; R.sub.3 being selected from the group consisting of alkyl,
trifluoromethyl, aryl, aralkyl, and methyl, at least one of said
alkyl, aryl, aralkyl, and methyl being optionally further
substituted by halogen; R.sub.4 being selected from the group
consisting of alkyl, cycloalkyl, heterocyclo alkyl, aryl,
heteroaryl, --(CH.sub.2).sub.n(OCH.sub.2CH.sub.2).sub.rR.sub.11,
--[CH.sub.2CH(OH)].sub.rCH.sub.2NR.sub.9R.sub.10 and
--(CH.sub.2).sub.nNR.sub.9R.sub.10, at least one said alkyl,
cylcoalkyl, heterocyclo alkyl, aryl and heteroaryl is optionally
further substituted by the group consisting of aryl, hydroxyl,
amino, carboxamido, alkoxyl, aryloxy, aminoalkyl, hydroxyalkyl,
heterocyclo alkyl, carboxyl, alkoxycarbonyl and --NR.sub.9R.sub.10;
when X is nitrogen, R.sub.5 is absent, R.sub.6, R.sub.7, R.sub.8
are each independently selected from the group consisting of
hydrogen and halogen; when X is carbon, R.sub.5, R.sub.6, R.sub.7,
R.sub.8 are each independently selected from the group consisting
of hydrogen, halogen, hydroxyalkyl, alkyl, cycloalkyl, heterocyclo
alkyl, aryl, heteroaryl, hydroxyl, cyano, nitro, --OR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9R.sub.10,
--(CH.sub.2).sub.nCO.sub.2R.sub.9,
--(CH.sub.2).sub.nCONR.sub.9R.sub.10, --COR.sub.9,
--NR.sub.9COR.sub.10, --SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10,
at least one of said aryl, heteroaryl, cycloalkyl and heterocyclo
alkyl is optionally further substituted by the group consisting of
alkyl, alkoxyl and halogen; R.sub.9 and R.sub.10 each being
independently selected from the group consisting of hydrogen,
alkyl, cycloalkyl, aryl, heterocyclo alkyl and heteroaryl, at least
one of said alkyl, cycloalkyl, aryl, heterocyclo alkyl and
heteroaryl being optionally further substituted by the group
consisting of alkyl, aryl, haloaryl, hydroxyl, amino, cyano,
alkoxyl, aryloxy, hydroxyalkyl, heterocyclo alkyl, carboxyl,
alkoxycarbonyl and --NR.sub.9R.sub.10; R.sub.9 and R.sub.10 being
taken together with the attached atom to form 4 to 8 membered
hetero rings, wherein said 4 to 8 membered hetero rings contain at
least one heteroatom selected from the group consisting of N, O and
S, and said 4 to 8 membered rings being optionally further
substituted by the group consisting of alkyl, halogen, aryl,
heteroaryl, haloalkyl, hydroxyl, cyano, alkoxyl, aryloxy,
aminoalkyl, hydroxyalkyl, heterocyclo alkyl, carboxyl,
alkoxycarbonyl and --NR.sub.9R.sub.10; R.sub.11 is selected from
the group consisting of hydrogen and alkyl; n is an integer from 2
to 6; z is an integer from 1 to 4; and r is an integer from 1 to
6.
2. The method according to claim 1, wherein said protein kinase is
selected from the group consisting of receptor tyrosine kinase,
non-receptor protein tyrosine kinase and serine-threonine protein
kinase.
3. The method according to claim 1, wherein R.sub.1 and R.sub.2 are
hydrogen.
4. The method according to claim 1, wherein said compound of
formula (I) has the formula (IA): ##STR00204## wherein R.sup.1 to
R.sup.8, and X are defined as those in claim 1.
5. The method according to claim 1, wherein said compound of
formula (I) has the formula (IB): ##STR00205## wherein R.sup.1 to
R.sup.8, and X are defined as those in claim 1.
6. The method according to claim 1, wherein said compound of
formula (I) is selected from the group consisting of: ##STR00206##
##STR00207## ##STR00208## ##STR00209## ##STR00210## ##STR00211##
##STR00212## ##STR00213## ##STR00214## ##STR00215## ##STR00216##
##STR00217## ##STR00218## ##STR00219## ##STR00220## ##STR00221##
##STR00222## ##STR00223## ##STR00224## ##STR00225##
##STR00226##
7. The method according to claim 1, wherein said pharmaceutical
composition comprises an effective amount of a compound of formula
(I) or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
8. The method according to claim 1, wherein said pharmaceutically
acceptable salt is salt formed with the acid selected from the
group consisting of malic acid, lactic acid, maleic acid,
hydrochloric acid, methanesulfonic acid, sulfuric acid, phosphoric
acid, citric acid, tartaric acid, acetic acid and trifluoroacetic
acid.
9. A method of treating a protein kinase related disorder
comprising a step of administering to a patient in need thereof a
therapeutically effective amount of the compound of formula (I):
##STR00227## or a pharmaceutically acceptable salt thereof, or a
pharmaceutical composition containing the same, wherein said
protein kinase related disorder is a non-colon cancer disorder; X
being selected from the group consisting of carbon and nitrogen;
R.sub.1 and R.sub.2 each being independently selected from the
group consisting of hydrogen and alkyl; R.sub.3 being selected from
the group consisting of alkyl, trifluoromethyl, aryl, aralkyl, and
methyl, at least one of said alkyl, aryl, aralkyl, and methyl being
optionally further substituted by halogen; R.sub.4 being selected
from the group consisting of alkyl, cycloalkyl, heterocyclo alkyl,
aryl, heteroaryl,
--(CH.sub.2).sub.n(OCH.sub.2CH.sub.2).sub.rR.sub.11,
--[CH.sub.2CH(OH)].sub.rCH.sub.2NR.sub.9R.sub.10 and
--(CH.sub.2).sub.nNR.sub.9R.sub.10, at least one said alkyl,
cylcoalkyl, heterocyclo alkyl, aryl and heteroaryl is optionally
further substituted by the group consisting of aryl, hydroxyl,
amino, carboxamido, alkoxyl, aryloxy, aminoalkyl, hydroxyalkyl,
heterocyclo alkyl, carboxyl, alkoxycarbonyl and --NR.sub.9R.sub.10;
when X is nitrogen, R.sub.5 is absent, R.sub.6, R.sub.7, R.sub.8
are each independently selected from the group consisting of
hydrogen and halogen; when X is carbon, R.sub.5, R.sub.6, R.sub.7,
R.sub.8 are each independently selected from the group consisting
of hydrogen, halogen, hydroxyalkyl, alkyl, cycloalkyl, heterocyclo
alkyl, aryl, heteroaryl, hydroxyl, cyano, nitro, --OR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9R.sub.10,
--(CH.sub.2).sub.nCO.sub.2R.sub.9,
--(CH.sub.2).sub.nCONR.sub.9R.sub.10, --COR.sub.9,
--NR.sub.9COR.sub.10, --SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10,
at least one of said aryl, heteroaryl, cycloalkyl and heterocyclo
alkyl is optionally further substituted by the group consisting of
alkyl, alkoxyl and halogen; R.sub.9 and R.sub.10 each being
independently selected from the group consisting of hydrogen,
alkyl, cycloalkyl, aryl, heterocyclo alkyl and heteroaryl, at least
one of said alkyl, cycloalkyl, aryl, heterocyclo alkyl and
heteroaryl being optionally further substituted by the group
consisting of alkyl, aryl, haloaryl, hydroxyl, amino, cyano,
alkoxyl, aryloxy, hydroxyalkyl, heterocyclo alkyl, carboxyl,
alkoxycarbonyl and --NR.sub.9R.sub.10; R.sub.9 and R.sub.10 being
taken together with the attached atom to form 4 to 8 membered
hetero rings, wherein said 4 to 8 membered hetero rings contain at
least one heteroatom selected from the group consisting of N, O and
S, and said 4 to 8 membered rings being optionally further
substituted by the group consisting of alkyl, halogen, aryl,
heteroaryl, haloalkyl, hydroxyl, cyano, alkoxyl, aryloxy,
aminoalkyl, hydroxyalkyl, heterocyclo alkyl, carboxyl,
alkoxycarbonyl and --NR.sub.9R.sub.10; R.sub.11 is selected from
the group consisting of hydrogen and alkyl; n is an integer from 2
to 6; z is an integer from 1 to 4; and r is an integer from 1 to
6.
10. The method according to claim 9, wherein said protein kinase
related disorder is selected from the group consisting of the
disorder related to VEGFR, EGFR, HER-2, HER-3, HER-4, PDGFR, c-Kit,
c-Met, FGFR, Ret, Flt1 and Flt3.
11. The method according to claim 10, wherein said protein kinase
related disorder is selected from the group consisting of the
disorder related to VEGFR, PDGFR, c-Kit, Ret, Flt1 and Flt3.
12. The method according to claim 10, wherein said protein kinase
related disorder is selected from the group consisting of leukemia,
diabetes, autoimmune diseases, hyperplasias, psoriasis,
osteoarthritis, rheumatoid arthritis, angiogenesis, cardiovascular
diseases, multiple angioblastoma, inflammatory diseases and
fibrosis.
13. The method according to claim 12, wherein said protein kinase
related disorder is cancer selected from squamous cell carcinoma,
renal cell carcinoma, Kaposi's sarcoma, non-small cell lung cancer,
small cell lung cancer, lymphoma, thyroid adenocarcinoma, breast
cancer, head and neck cancer, uterine cancer, esophageal cancer,
melanoma, bladder cancer, carcinosarcoma in urinary and genital
system, gastrointestinal carcinoma, gliomas, colorectal cancer and
ovarian cancer.
14. The method according to claim 9, wherein R.sub.1 and R.sub.2
are hydrogen.
15. The method according to claim 9, wherein said compound of
formula (I) has the formula (IA): ##STR00228## wherein R.sup.1 to
R.sup.8, and X are defined as those in claim 9.
16. The method according to claim 9, wherein said compound of
formula (I) has the formula (IB): ##STR00229## wherein R.sup.1 to
R.sup.8, and X are defined as those in claim 9.
17. The method according to claim 9, wherein said compound of
formula (I) is selected from the group consisting of: ##STR00230##
##STR00231## ##STR00232## ##STR00233## ##STR00234## ##STR00235##
##STR00236## ##STR00237## ##STR00238## ##STR00239## ##STR00240##
##STR00241## ##STR00242## ##STR00243## ##STR00244## ##STR00245##
##STR00246## ##STR00247## ##STR00248## ##STR00249##
##STR00250##
18. The method according to claim 9, wherein said pharmaceutical
composition comprises an effective amount of a compound of formula
(I) or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
19. The method according to claim 9, wherein said pharmaceutically
acceptable salt is salt formed with the acid selected from the
group consisting of malic acid, lactic acid, maleic acid,
hydrochloric acid, methanesulfonic acid, sulfuric acid, phosphoric
acid, citric acid, tartaric acid, acetic acid and trifluoroacetic
acid.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a continuation-in-part patent
application of U.S. patent application Ser. No. 12/451,466, filed
on Dec. 3, 2009, which is a national stage application of
International Patent Application No. PCT/CN2008/001352, filed on
May 14, 2008, published as WO 2008/138232 on Nov. 20, 2008; which
claims priority to Chinese Patent Application No. 200710107463.8,
filed on May 14, 2007; and Chinese Patent Application No.
200810087564.8, filed on Apr. 11, 2008, each of which is
incorporated by reference herein.
FIELD OF THE INVENTION
[0002] The invention relates to a method of inhibiting the
catalytic activity of a protein kinase comprising contacting said
protein kinase with pyrrolo-nitrogenous heterocyclic derivatives,
or a pharmaceutically acceptable salt thereof, or pharmaceutical
compositions containing such derivatives, and also relates to a
method of treating a protein kinase related disorder comprising a
step of administering to a patient in need thereof a
therapeutically effective amount of pyrrolo-nitrogenous
heterocyclic derivatives, or a pharmaceutically acceptable salt
thereof, or pharmaceutical compositions containing such
derivatives.
BACKGROUND OF THE INVENTION
[0003] Cellular signal transduction is a fundamental mechanism
whereby extracellular stimuli are relayed to the interior of cells
and subsequently regulate diverse cellular processes. These signals
regulate a wide variety of physical responses in the cell including
proliferation, differentiation, apoptosis and motility. The
extracellular signals take the form of a diverse variety of soluble
factors including growth factors as well as paracrine, autocrine
and endocrine factors. By binding to specific transmembrane
receptors, growth factor ligands communicate extracellular signals
to the intracellular signalling pathways, thereby causing the
individual cell to respond to extracellular signals. Many of these
signal transduction processes utilize the reversible process of the
phosphorylation of proteins involving specific protein kinases and
phosphatases.
[0004] Protein kinases (PKs) are enzymes that catalyze the
phosphorylation of hydroxyl groups on tyrosine, serine and
threonine residues of proteins, whereas protein phosphatases
hydrolyze phosphate moieties from phosphorylated protein
substrates. The converse functions of protein kinases and protein
phosphatases balance and regulate the flow of signals in signal
transduction processes. The phosphorylation state of a protein can
affect its conformation, enzymatic activity, and cellular location,
is modified through the reciprocal actions of protein kinases and
protein phosphatases. Phosphorylation is an important regulatory
mechanism in the signal transduction process and aberrations in the
process result in abnormal cell differentiation, transformation and
growth. For example, it has been discovered that a cell may become
cancerous by virtue of the transformation of a portion of its DNA
into an oncogene. Several such oncogenes encode proteins which are
receptors for growth factors, for example tyrosine kinases.
Tyrosine kinases may also be mutated to constitutively active forms
that result in the transformation of a variety of human cells.
Alternatively, the overexpression of normal tyrosine kinase enzymes
may also result in abnodal cell proliferation.
[0005] There are two classes of PKs, the protein tyrosine kinases
(PTKs) and the serine-threonine kinases (STKs). PTKs phosphorylate
tyrosine residue on a protein. STKs phosphorylate serine or/and
threonine on a protein. Tyrosine kinases can be of not only the
receptor-type (having extracellular, transmembrane and
intracellular domains) but the non-receptor type (being wholly
intracellular). One of the prime aspects of PTK activity is their
involvement with growth factor receptors which are cell-surface
proteins. Growth factor receptors with PTK activity are known as
receptor tyrosine kinases ("RTKs"). About 90 tyrosine kinases have
been identified in the human genome, of which about 60 are of the
receptor type and about 30 are of the non-receptor type. These can
be categorized into 20 receptor tyrosine kinase sub-families
according to the families of growth factors that they bind and into
10 non-receptor tyrosine kinase sub-families (Robinson et al,
Oncogene, 2000, 19, 5548-5557).
[0006] The Receptor Tyrosine Kinases (RTKs) family includes: (1)
the EGF family of receptor tyrosine kinases such as the EGF,
TGF.alpha., Neu and erbB receptors; (2) the insulin family of
receptor tyrosine kinases such as the insulin and IGF1 receptors
and insulin-related receptor (IRR); (3) the Class III family of
receptor tyrosine kinases such as the platelet-derived growth
factor (PDGF) receptor tyrosine kinases, for example the
PDGF.alpha. and PDGF.beta. receptors, the stem cell factor receptor
tyrosine kinase SCF RTK (commonly known as c-Kit), the fms-related
tyrosine kinase 3 (Flt3) receptor tyrosine kinase and the
colony-stimulating factor 1 receptor (CSF-1R) tyrosine kinase and
the like. They play critical role in the control of cell growth and
differentiation and are key mediators of cellular signals leading
to the production of growth factors and cytokines (Schlessinger and
Ullrich, Neuron 1992, 9, 383). A partial, non-limiting, list of
such kinases includes Abl, ARaf, ATK, ATM, bcr-abl, Blk, BRaf, Brk,
Btk, CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CHK,
AuroraA, AuroraB, AuroraC, cfms, c-fms, c-Kit, c-Met, cRaf1, CSF1R,
CSK, c-Src, EGFR, ErbB2, ErbB3, ErbB4, ERK, ERK1, ERK2, Fak, fes,
FGFR1, FGFR2, FGFR3, FGFR4, FGFR5, Fgr, FLK-4, Fps, Frk, Fyn, GSK,
gsk3a, gsk3b, Hck, Chk, Axl, Pim-1, Plh-1, IGF-1R, IKK, IKK1, IKK2,
IKK3, INS-R, Integrin-linked kinase, Jak, JAK1, JAK2, JAK3, JNK,
JNK, Lck, Lyn, MEK, MEK1, MEK2, p38, PDGFR, PIK, PKB1, PKB2, PKB3,
PKC, PKCa, PKCb, PKCd, PKCe, PKCg, PKC1, PKCm, PKCz, PLK1,
Polo-like kinase, PYK2, tie.sub.1, tie.sub.2, TrkA, TrkB, TrkC,
UL13, UL97, VEGF-R1, VEGF-R2, Yes and Zap70 and the like. Protein
kinases have also been implicated as targets in central nervous
system disorders such as Alzheimer's (Mandelkow, E. M. et al. FEBS
Lett. 1992, 314, 315; Sengupta, A. et al. Mol. Cell. Biochem. 1997,
167, 99), pain sensation (Yashpal, K. J. Neurosci. 1995, 15,
3263-72), inflammatoy disorders such as arthritis (Badger, J.
Pharmn Exp. Ther. 1996, 279, 1453), psoriasis (Dvir, et al. J. Cell
Biol. 1991, 113, 857), bone diseases such as osteoporosis (Tanaka
et al, Nature, 1996, 383, 528), cancer (Hunter and Pines, Cell
1994, 79, 573), atherosclerosis (Hajjar and Pomerantz, FASEB J.
1992, 6, 2933), thrombosis (Salari, FEBS 1990, 263, 104), metabolic
disorders such as diabetes (Borthwick, A. C. et al. Biochem.
Biophys. Res. Commun. 1995, 210, 738), blood vessel proliferative
disorders such as angiogenesis (Strawn et al. Cancer Res. 1996, 56,
3540; Jackson et al. J. Pharm. Exp. Ther. 1998, 284, 687),
autoimmune diseases and transplant rejection (Bolen and Brugge,
Ann. Rev. Immunol. 1997, 15, 371) and infection diseases such as
viral (Littler, E. Nature 1992, 358, 160), and fungal infections
(Lum, R. T. PCT Int Appl., WO 9805335 A1 980212).
[0007] RTKs mediated signal transduction is initiated by
extracellular interaction with a specific growth factor (ligand),
followed by receptor dimerization, transient stimulation of the
intrinsic protein tyrosine kinase activity and phosphorylation.
Binding sites are thereby created for intracellular signal
transduction molecules and lead to the formation of complexes with
a spectrum of cytoplasmic signaling molecules that facilitate the
appropriate cellular responses, e.g., cell division
(multiplication), and responses to the extracellular
microenvironment.
[0008] With respect to receptor tyrosine kinases, it has been shown
also that tyrosine phosphorylation sites function as high-affinity
binding sites for SH2 (src homology) domains of signaling
molecules. Several intracellular substrate proteins that associate
with receptor tyrosine kinases have been identified. They may be
divided into two principal groups: (1) substrates which have a
catalytic domain; and (2) substrates which lack such domain but
serve as adapters and associate with catalytically active
molecules. The specificity of the interactions between receptors or
proteins and SH2 domains of their substrates is determined by the
amino acid residues immediately surrounding the phosphorylated
tyrosine residue. Differences in the binding affinities between SH2
domains and the amino acid sequences surrounding the
phosphotyrosine residues on particular receptors are consistent
with the observed differences in their substrate phosphorylation
profiles. These observations suggest that the function of each
receptor tyrosine kinase is determined not only by its pattern of
expression and ligand availability but also by the array of
downstream signal transduction pathways that are activated by a
particular receptor. Thus, phosphorylation provides an important
regulatory step which determines the selectivity of signaling
pathways recruited by specific growth factor receptors, as well as
differentiation factor receptors. Aberrant expression or mutations
in the protein tyrosine kinases have been shown to lead to either
uncontrolled cell proliferation (e.g. malignant tumor growth) or to
defects in key developmental processes.
[0009] It has been identified that such mutated and overexpressed
forms of tyrosine kinases are present in a large proportion of
common human cancers such as the leukaemia, breast cancer, prostate
cancer, non-small cell lung cancer (NSCLC) including
adenocarcinomas and squamous cell cancer of the lung,
gastrointestinal cancer including colon, rectal and stomach cancer,
bladder cancer, oesophageal cancer, ovarian cancer and pancreatic
cancer and the like. As further human tumour tissues are tested, it
is expected that the widespread prevalence and relevance of
tyrosine kinases will be further established. For example, it has
been shown that EGFR tyrosine kinase is mutated and overexpressed
in several human cancers including in tumours of the lung, head and
neck, gastrointestinal tract, breast, oesophagus, ovary, uterus,
bladder and thyroid.
[0010] One subfamily designated the "HER" or "Erb" RTKs, which
include EGFR (epithelial growth factor receptor), HER2, HER3 and
HER4. These RTKs consist of an extracellular glycosylated ligand
binding domain, a transmembrane domain and an intracellular
cytoplasm catalytic domain that can phosphorylate tyrosine residues
on proteins. The enzymatic activity of receptor tyrosine kinases
can be stimulated by either overexpression of receptor, or by
ligand-mediated dimerization. The formation of homodimers as well
as heterodimers has been demonstrated for the HER2 receptor family.
An example of homodimerization is the dimerization of HER1 (EGF
receptor) by the EGF family of ligands (which includes EGF,
transforming growth factor alpha, betacellulin, heparin-binding
EGF, and epiregulin). Heterodimerization among the four HER
receptor kinases can be promoted by binding to members of the
heregulin (also referred to neuregulin) family of ligands. Such
heterodimerization as involving HER2 and HER3, or a HER3 and HER4
combination, results in a significant stimulation of the tyrosine
kinase activity of the receptor dimers even though one of the
receptors (HER3) is enzymatically inert. The kinase activity of
HER2 has been shown to be activated also by virtue of
overexpression of the receptor alone in a variety of cell types.
Activation of receptor homodimers and heterodimers results in
phosphorylation of tyrosine residues on the receptors and on other
intracellular proteins. This is followed by the activation of
intracellular signaling pathways such as those involving the
microtubule associated protein kinase (MAP kinase) and the
phosphatidylinositol3-kinase (PI3 kinase). Activation of these
pathways has been shown to lead to cell proliferation and the
inhibition of apoptosis.
[0011] Another RTK subfamily includes insulin receptor (IR),
insulin-like growth factor I receptor (IGF-1R) and insulin receptor
related receptor (IRR). IR and IGF-1R interact with insulin, IGF-I
and IGF-.quadrature. to form a heterotetramer of two entirely
extracellular glycosylated a subunits and two .beta. subunits which
cross the cell membrane and which contain the tyrosine kinase
domain.
[0012] A third RTK subfamily is referred to as the platelet derived
growth factor receptor (PDGFR) group, which includes PDGFR.alpha.,
PDGFR.beta., CSFIR, c-Kit and c-fms. These receptors consist of
glycosylated extracellular domains composed of variable members of
immunoglobin-like loops and an intracellular domain wherein the
tyrosine kinase domain is interrupted by unrelated amino acid
sequences.
[0013] Platelet derived growth factor receptors such as
PDGFR.alpha. and PDGFR are also transmembrance tyrosine kinase
receptors. Upon binding of the ligand, they form either homodimers
(PDGF-AA, PDGF-BB) or heterodimers (PDGF-AB). Follow is the
receptor dimerizes, its tyrosine kinase is activated. This leads to
downstream signaling and thus may support tumor growth. Mutations
in this gene allow for receptor activation independent of ligand
binding and appear to be driving forces in oncogenesis. An
expression of PDGF, the growth factor that activates PDGFR, was
observed in a number of different tumor cell lines, inter alia in
mamma, colon, ovarian, prostate carcinoma, sarcoma and
glioblastomas cell lines. Among the tumors, brain tumors and
prostate carcinoma (including adenocarcinomas and bone metastasis)
have found special interest. Interesting data also exist regarding
malign gliomes.
[0014] c-Kit is a tyrosine kinase receptor which belongs to the
PDGF receptor family and becomes activated upon binding of its
ligand SCF (stem-cell factor). The expression pattern of c-Kit has
been studied e.g. in a panel of different primary solid tumors. A
strong expression of c-Kit could be found inter alia in sarcoma,
gastrointestinal stromal tumors (GIST), seminoma and carcinoids
[Weber et al., J. Clin. Oncol. 22(14S), 9642 (2004)]. GISTs are
non-epithelial tumors. Many occur in the stomach, less in the small
intestine and still less in the esophagus. Dissemination to the
liver, omentum and peritoneal cavity can be observed. GISTS
probably arise from Interstitial Cajal Cells (ICC) which normally
form part of the autonomic nervous system of the intestine and take
part in the control of motility. Most (50 to 80%) of GISTS arise
due to c-Kit gene mutation. In the gut, a staining positive for
c-Kit/CD117 is likely to be a GIST. Mutations of c-Kit can make
c-Kit function independent of activation by SCF, leading to a high
cell division rate and possibly genomic instability. Also in mast
cell tumors aberrations of c-Kit could be observed, as well as in
mastocytosis and associated myeloproliferative syndrome and
Urticaria Pigmentosa. An expression and/or aberrations of c-Kit can
also be found in acute myeloicanemia (AML) and malign lymphomas. A
c-Kit expression can also be demonstrated in small cell bronchial
carcinoma, seminomas, dysgerminomas, testicular intraepithelial
neoplasias, melanomas, mamma carcinomas, neuroblastomas, Ewing
sarcoma, some soft part sarcomas as well as papillary/follicular
thyroid carcinoma (see Sch{hacek over (u)}tte et al., innovartis
3/2001). Inherited mutations of the RET (rearranged during
transfection) proto-oncogene are e.g. known to be tumorigenic in
patients with multiple endocrine neoplasia type 2 (MEN 2) which may
lead to pheochromocytoma, medullary thyroid carcinoma and
parathyroid hyperplasia/adenoma (see Huang et al., Cancer Res. 60,
6223-6 (2000)).
[0015] Another group which, because of its similarity to the PDGFR
subfamily, is sometimes subsumed into the later group is the fetus
liver kinase (Flk) receptor subfamily. This group is believed to be
made up of kinase insert domain-receptor fetal liver kinase-1
(KDR/FLK-1, VEGFR2), Flk-1R, Flk-4 and Fms-like tyrosine kinase 1
(Flt-1).
[0016] A further member of the tyrosine kinase growth factor
receptor family is the fibroblast growth factor (FGF) receptor
subgroup. This group consists of four receptors, FGFR1-4, seven
ligands, and FGF1-7. While not yet well defined, it appears that
the receptors consist of a glycosylated extracellular domain
containing a variable number of immunoglobin-like loops and an
intracellular domain in which the tyrosine kinase sequence is
interrupted by regions of unrelated amino acid sequences.
[0017] Still another member of the tyrosine kinase growth factor
receptor family is the vascular endothelial growth factor (VEGF)
receptor subgroup, VEGF is a dimeric glycoprotein similar to PDGF
but has different biological functions and target cell specificity
in vivo. In particular, VEGFRs are known to be involved in the
control of the onset of angiogenesis. As especially solid tumors
depend on good blood supply, inhibition of VEGFRs and thus
angiogenesis is under clinical investigation in the treatment of
such tumors, showing promising results. VEGF is also a major player
in leukemias and lymphomas and highly expressed in a variety of
solid malignant tumors, correlating well with malignant disease
progression. Examples of tumor diseases with VEGFR-2 (KDR)
expression are lung carcinomas, breast carcinomas, Non Hodgkin's
lymphomas, ovarian carcinoma, pancreatic cancer, malignant pleural
mesothelioma and melanoma. In addition to its angiogenic activity,
the ligand of VEGFR, VEGF, may promote tumor growth by direct
pro-survival effects in tumor cells. PDGF is also involved in
angiogenesis, the process of forming new blood vessels that is
critical for continuing tumor growth. Normally, angiogenesis plays
an important role in processes such as embryonic development, wound
healing and several components of female reproductive function.
However, undesirable or pathological angiogenesis has been
associated with a number of disease states including diabetic
retinopathy, psoriasis, cancer, rheumatoid arthritis, atheroma,
Kaposi's sarcoma and haemangioma. Angiogenesis is stimulated via
the promotion of the growth of endothelial cells. Several
polypeptides with in vitro endothelial cell growth promoting
activity have been identified including acidic and basic fibroblast
growth factors (aFGF and bFGF) and vascular endothelial growth
factor (VEGF). By virtue of the restricted expression of its
receptors, the growth factor activity of VEGF, in contrast to that
of aFGF and bFGF, is relatively specific towards endothelial cells.
Recent evidence indicates that VEGF is an important stimulator of
both normal and pathological angiogenesis and vascular
permeability. This cytokine induces a vascular sprouting phenotype
by inducing endothelial cell proliferation, protease expression and
migration which subsequently leads to the formation of capillary
tubes that promote the formation of the hyperpermeable, immature
vascular network which is the characteristic of pathological
angiogenesis. Accordingly, antagonism of the activity of VEGF is
expected to be beneficial in the treatment of a number of disease
states that are associated with angiogenesis or increased vascular
permeability such as cancer, especially in inhibiting the
development of tumors.
[0018] FLT3 (fms-like tyrosine kinase) is a member of the type Ill
receptor tyrosine kinase (RTK) family. Aberrant expression of the
FLT3 gene has inter alia been documented in both adult and
childhood leukemias including acute myeloid leukemia (AML), AML
with trilineage myelodysplasia (AML/TMDS), acute lymphoblastic
leukemia (ALL), and myelodysplastic syndrome (MDS), as well as MILL
(mixed-lineage leukemia). Activating mutations of the FLT3 receptor
have been found in about 35% of patients with acute myeloblastic
leukemia (AML), and are associated with a poor prognosis. The most
common mutation involves an in-frame duplication within the
juxtamembrane domain, with an additional 5-10% of patients having a
point mutation at asparagine 835. Both of these mutations are
associated with constitutive activation of the tyrosine kinase
activity of FLT3, and result in proliferation and viability signals
in the absence of ligand. Patients expressing the mutant form of
the receptor have been shown to have a decreased chance for cure.
Thus, there is accumulating evidence for a role for hyperactivated
(mutated) FLT3 kinase activity in human leukemias and
myelodysplastic syndrome.
[0019] The hepatocyte growth factor (HGF) receptor (c-MET or HGFR)
receptor tyrosine kinase (RTK) has been shown in many human cancers
to be involved in oncogenesis, tumor progression with enhanced cell
motility and invasion, as well as metastasis (see, Ma, P. C. et al.
(2003b). Cancer Metastasis Rev, 22, 309-25; Maulik, G. et al.
(2002b). Cytokine Growth Factor Rev, 13, 41-59). c-MET (HGFR) can
be activated through overexpression or mutations in various human
cancers including small cell lung cancer (SCLC) (Ma, P. C. et al.
(2003a). Cancer Res, 63, 6272-6281).
[0020] c-MET is a receptor tyrosine kinase that is encoded by the
Met proto-oncogene and transduces the biological effects of
hepatocyte growth factor (HGF). It is a transmembrane glycoprotein
with tyrosine kinase activity, which contribute to multi-cell
multiplication and division. The c-Met proto-oncogene is
overexpressed in numerous human malignancy especially in thyroid
tumour, which is closely related with pathologic staging, tumour
invasion and metastasis.
[0021] A more complete listing of the known RTK subfamilies is
described in Plowman et al., DN&P 7(6): 334-339 (1994) which is
incorporated by reference, as if fully set forth herein.
[0022] Besides PTKs, additional cell enzyme family is present,
called as receptor tyrosine kinases inhibitor (abbreviated as
"CTK"). At present, over twenty-four CTKs, comprising eleven
subfamilies (Src, Frk, Btk, Csk, Abl, Zap70, Fes/Fps, Fak, Jak, Ack
and LIMK) have been identified. At present, the Src subfamily of
CTKs are comprised of the largest number of PTKs and include Src,
Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk. The Src subfamily of
enzymes has been linked to oncogenesis. A more detailed discussion
of CTKs is provided in Bolen, 1993, Oncogen 8: 2025-2031, which is
incorporated herein by reference, including any drawings.
[0023] The serine/threonine kinases, or STKs, like the CTKs, are
predominantly intracellular although there are a few receptor
kinases of the STK type. STKs are the most common of the cytosolic
kinases; i.e., kinases that perform their function in that part of
the cytoplasm other than the cytoplasmic organelles and
cytoskelton. The cytosol is the region within the cell where much
of the cell's intermediary metabolic and biosynthetic activity
occurs; e.g., it is in the cytosol that proteins are synthesized on
ribosomes.
[0024] A further characteristic of hyperproliferative diseases such
as cancer is damage to the cellular pathways that control progress
through the cell cycle which, in normal eukaryotic cells, involves
an ordered cascade of protein phosphorylation. As for signal
transduction mechanisms, several families of protein kinases appear
to play critical roles in the cell cycle cascade.
[0025] With regard to cancer, two of the major hypotheses advanced
to explain the excessive cellular proliferation that drives tumor
development relate to functions known to be PK regulated. That is,
it has been suggested that malignant cell growth results from a
breakdown in the mechanisms that control cell division or
differentiation. It has been shown that the protein products of a
number of proto-oncogenes are involved in the signal transduction
pathways that regulate cell growth and differentiation. These
protein products of proto-oncogenes include the extracellular
growth factors, transmembrane growth factor PTK receptors (RTKs),
cytoplasmic PTKs (CTKs) and cytosolic STKs, discussed above.
[0026] There is a need for novel viable small-molecule inhibitors
which posess anti-tumor cell proliferative activities. These small
molecules are expected to inhibit one or more RTKs, CTKs or STKs,
and are useful in treating or ameliorating RTKs, CTKs or STKs
mediated, angiogenesis mediated or hyperproliferative disorder.
[0027] Based on the structure of tyrosine kinase inhibitor SU-11248
and the pyrrolofused heterocycle Formula (X) which showed great
bioactivities reported by the patent (U.S. Pat. No. 6,599,902B2),
the present invention is directed to design and synthesize the
pyrrolofused multiple-membered aza-heterocyclic derivatives and has
got a better pharmacological data. In order to improve the
pharmacokinetics profile of pyrrolofused multiple-membered
aza-heterocyclic derivatives, the present invention is directed to
design compounds of formula (I). The compounds of the invention
have obvious structure differences with the existing compounds in
prior art, and they also show more efficiency and more
function.
##STR00002##
SUMMARY OF THE INVENTION
[0028] In order to overcome the deficiency of the prior art, the
present invention is directed to provide a method of inhibiting the
catalytic activity of a protein kinase comprising contacting said
protein kinase with pyrrolo-nitrogenous heterocyclic derivative
having formula (I):
##STR00003##
its tautomer, enantiomer, diastereomer, racemate, or
pharmaceutically acceptable salt thereof, and metabolite, precursor
or prodrug thereof, or pharmaceutically acceptable salt thereof, or
a pharmaceutical composition containing the same,
Wherein:
[0029] X is selected from the group consisting of carbon and
nitrogen;
[0030] R.sub.1 and R.sub.2 are each independently selected from the
group consisting of hydrogen and alkyl;
[0031] R.sub.3 is selected from the group consisting of alkyl,
trifluoromethyl, aryl and aralkyl, wherein said alkyl, aryl or
aralkyl is optionally further substituted by halogen;
[0032] R.sub.4 is selected from the group consisting of alkyl,
cycloalkyl, heterocyclo alkyl, aryl, heteroaryl,
--(CH.sub.2).sub.n(OCH.sub.2CH.sub.2).sub.rR.sub.11,
--[CH.sub.2CH(OH)].sub.rCH.sub.2NR.sub.9R.sub.10 and
--(CH.sub.2).sub.nNR.sub.9R.sub.10, wherein said alkyl, cylcoalkyl,
heterocyclo alkyl, aryl or heteroaryl is optionally further
substituted by the group selected from the group consisting of
aryl, hydroxyl, amino, carboxamido, alkoxyl, aryloxy, aminoalkyl,
hydroxyalkyl, heterocyclo alkyl, carboxyl, alkoxycarbonyl and
--NR.sub.9R.sub.10;
[0033] when X is nitrogen, R.sub.5 is absent, R.sub.6, R.sub.7,
R.sub.8 are each independently selected from the group consisting
of hydrogen and halogen;
[0034] when X is carbon atom, R.sub.5, R.sub.6, R.sub.7, R.sub.8
are each independently selected from the group consisting of
hydrogen, halogen, hydroxyalkyl, alkyl, cycloalkyl, heterocyclo
alkyl, aryl, heteroaryl, hydroxyl, cyano, nitro, --OR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9R.sub.10,
--(CH.sub.2).sub.nCO.sub.2R.sub.9,
--(CH.sub.2).sub.nCONR.sub.9R.sub.10, --COR.sub.9,
--NR.sub.9COR.sub.10, --SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10,
wherein said aryl, heteroaryl, cycloalkyl or heterocyclo alkyl is
optionally further substituted by the group consisting of alkyl,
alkoxyl and halogen;
[0035] R.sub.9 and R.sub.10 are each independently selected from
the group consisting of hydrogen, alkyl, cycloalkyl, aryl,
heterocyclo alkyl and heteroaryl, wherein said alkyl, cycloalkyl,
aryl, heterocyclo alkyl or heteroaryl is optionally further
substituted by the group consisting of alkyl, aryl, haloaryl,
hydroxyl, amino, cyano, alkoxyl, aryloxy, hydroxyalkyl, heterocyclo
alkyl, carboxyl, alkoxycarbonyl and --NR.sub.9R.sub.10;
[0036] R.sub.9 and R.sub.10 are taken together with the attached
atom to form 4 to 8 membered hetero rings, wherein said 4 to 8
membered hetero rings may contain one to more heteroatoms selected
from the group consisting of N, O and S atom, and said 4 to 8
membered rings are optionally further substituted by the group
consisting of alkyl, halogen, aryl, heteroaryl, haloalkyl,
hydroxyl, cyano, alkoxyl, aryloxy, aminoalkyl, hydroxyalkyl,
heterocyclo alkyl, carboxyl, alkoxycarbonyl and
--NR.sub.9R.sub.10;
[0037] R.sub.11 is selected from the group consisting of hydrogen
and alkyl;
[0038] n is an integer from 2 to 6;
[0039] z is an integer from 1 to 4; and
[0040] r is an integer from 1 to 6;
[0041] or pharmaceutically acceptable salts thereof.
[0042] The method of inhibiting the catalytic activity of a protein
kinase, wherein said protein kinase is selected from the group
consisting of receptor tyrosine kinases, non-receptor protein
tyrosine kinases and serine-threonine protein kinases.
[0043] The method of inhibiting the catalytic activity of a protein
kinase, wherein R.sub.3 is preferably methyl.
[0044] The method of inhibiting the catalytic activity of a protein
kinase, wherein R.sub.1 and R.sub.2 are preferably hydrogen.
[0045] The method of inhibiting the catalytic activity of a protein
kinase, wherein said compound of formula (I) has the formula
(IA):
##STR00004##
Wherein:
[0046] X is selected from the group consisting of carbon and
nitrogen;
[0047] R.sub.1 and R.sub.2 are each independently selected from the
group consisting of hydrogen and alkyl;
[0048] R.sub.3 is selected from the group consisting of alkyl,
trifluoromethyl, aryl and aralkyl, wherein said alkyl, aryl or
aralkyl is optionally further substituted by halogen;
[0049] R.sub.4 is selected from the group consisting of alkyl,
cycloalkyl, heterocyclo alkyl, aryl, heteroaryl,
--(CH.sub.2).sub.n(OCH.sub.2CH.sub.2).sub.rR.sub.11,
--[CH.sub.2CH(OH)].sub.rCH.sub.2NR.sub.9R.sub.10 and
--(CH.sub.2).sub.nNR.sub.9R.sub.10, wherein said alkyl, cylcoalkyl,
heterocyclo alkyl, aryl or heteroaryl is optionally further
substituted by the group consisting of aryl, hydroxyl, amino,
carboxamido, alkoxyl, aryloxy, aminoalkyl, hydroxyalkyl,
heterocyclo alkyl, carboxyl, alkoxycarbonyl and
--NR.sub.9R.sub.10;
[0050] when X is nitrogen, R.sub.5 is absent, R.sub.6, R.sub.7,
R.sub.8 are each independently selected from the group consisting
of hydrogen and halogen;
[0051] when X is carbon, R.sub.5, R.sub.6, R.sub.7, R.sub.8 are
each independently selected from the group consisting of hydrogen,
halogen, hydroxyalkyl, alkyl, cycloalkyl, heterocyclo alkyl, aryl,
heteroaryl, hydroxyl, cyano, nitro, --OR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9R.sub.10,
--(CH.sub.2).sub.nCO.sub.2R.sub.9,
--(CH.sub.2).sub.nCONR.sub.9R.sub.10, --COR.sub.9,
--NR.sub.9COR.sub.10, --SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10,
wherein said aryl, heteroaryl, cycloalkyl or heterocyclo alkyl is
optionally further substituted by the group consisting of alkyl,
alkoxyl and halogen;
[0052] R.sub.9 and R.sub.10 are each independently selected from
the group consisting of hydrogen, alkyl, cycloalkyl, aryl,
heterocyclo alkyl and heteroaryl, wherein said alkyl, cycloalkyl,
aryl, heterocyclo alkyl or heteroaryl is optionally further
substituted by the group consisting of alkyl, aryl, haloaryl,
hydroxyl, amino, cyano, alkoxyl, aryloxy, hydroxyalkyl, heterocyclo
alkyl, carboxyl, alkoxycarbonyl and --NR.sub.9R.sub.10;
[0053] R.sub.9 and R.sub.10 are taken together with the attached
atom to form 4 to 8 membered hetero rings, wherein said 4 to 8
membered hetero rings may contain one or more heteroatoms selected
from the group consisting of N, O and S atom, and said 4 to 8
membered rings are optionally further substituted by the group
consisting of alkyl, halogen, aryl, heteroaryl, haloalkyl,
hydroxyl, cyano, alkoxyl, aryloxy, aminoalkyl, hydroxyalkyl,
heterocyclo alkyl, carboxyl, alkoxycarbonyl and
--NR.sub.9R.sub.10;
[0054] R.sub.11 is selected from the group consisting of hydrogen
and alkyl;
[0055] n is an integer from 2 to 6; and
[0056] r is an integer from 1 to 6;
[0057] or pharmaceutically acceptable salts thereof.
[0058] The method of inhibiting the catalytic activity of a protein
kinase, wherein said compound of formula (I) has the formula
(IB):
##STR00005##
Wherein:
[0059] X is selected from the group consisting of carbon and
nitrogen;
[0060] R.sub.1 and R.sub.2 are each independently selected from the
group consisting of hydrogen and alkyl;
[0061] R.sub.3 is selected from the group consisting of alkyl,
trifluoromethyl, aryl and aralkyl, wherein said alkyl, aryl or
aralkyl is optionally further substituted by halogen;
[0062] R.sub.4 is selected from the group consisting of alkyl,
cycloalkyl, heterocyclo alkyl, aryl, heteroaryl,
--(CH.sub.2).sub.n(OCH.sub.2CH.sub.2).sub.rR.sub.11,
--[CH.sub.2CH(OH)].sub.rCH.sub.2NR.sub.9R.sub.10 and
--(CH.sub.2).sub.nNR.sub.9R.sub.10, wherein said alkyl, cylcoalkyl,
heterocyclo alkyl, aryl or heteroaryl is optionally further
substituted by the group consisting of aryl, hydroxyl, amino, amide
group, carboxamido, alkoxyl, aryloxy, aminoalkyl, hydroxyalkyl,
heterocyclo alkyl, carboxyl, alkoxycarbonyl and
--NR.sub.9R.sub.10;
[0063] when X is nitrogen, R.sub.5 is absent, R.sub.6, R.sub.7,
R.sub.8 are each independently selected from the group consisting
of hydrogen and halogen;
[0064] when X is carbon, R.sub.5, R.sub.6, R.sub.7, R.sub.8 are
each independently selected from hydrogen, halogen, hydroxyalkyl,
alkyl, cycloalkyl, heterocyclo alkyl, aryl, heteroaryl, hydroxyl,
cyano, nitro, --OR.sub.9, --O[CH.sub.2CH.sub.2O].sub.rR.sub.11,
--NR.sub.9R.sub.10, --(CH.sub.2).sub.nCO.sub.2R.sub.9,
--(CH.sub.2).sub.nCONR.sub.9R.sub.10, --COR.sub.9,
--NR.sub.9COR.sub.10, --SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10,
wherein said aryl, heteroaryl, cycloalkyl or heterocyclo alkyl is
optionally further substituted by the group consisting of alkyl,
alkoxyl and halogen;
[0065] R.sub.9 and R.sub.10 are each independently selected from
the group consisting of hydrogen, alkyl, cycloalkyl, aryl,
heterocyclo alkyl and heteroaryl, wherein said alkyl, cycloalkyl,
aryl, heterocyclo alkyl or heteroaryl further is optionally
substituted by the group consisting of alkyl, aryl, haloaryl,
hydroxyl, amino, cyano, alkoxyl, aryloxy, hydroxyalkyl, heterocyclo
alkyl, carboxyl, alkoxycarbonyl and --NR.sub.9R.sub.10;
[0066] R.sub.9 and R.sub.10 are taken together with the attached
atom to form 4 to 8 membered hetero rings, wherein said 4 to 8
membered hetero rings may further optionally contain one or more
heteroatoms selected from the group consisting of N, O and S atom,
and said 4 to 8 membered rings are optionally further substituted
by the group consisting of alkyl, halogen, aryl, heteroaryl,
haloalkyl, hydroxyl, cyano, alkoxyl, aryloxy, aminoalkyl,
hydroxyalkyl, heterocyclo alkyl, carboxyl, alkoxycarbonyl and
--NR.sub.9R.sub.10;
[0067] R.sub.11 is selected from the group consisting of hydrogen
and alkyl;
[0068] n is an integer from 2 to 6; and
[0069] r is an integer from 1 to 6;
[0070] or pharmaceutically acceptable salts thereof.
[0071] The method of inhibiting the catalytic activity of a protein
kinase, wherein said pharmaceutical composition comprises an
effective amount of a compound of formula (I) or a pharmaceutically
acceptable salt thereof and a pharmaceutically acceptable
carrier.
[0072] The method of inhibiting the catalytic activity of a protein
kinase, wherein said pharmaceutically acceptable salt is salt
formed with the acid selected from the group consisting of malic
acid, lactic acid, maleic acid, hydrochloric acid, methanesulfonic
acid, sulfuric acid, phosphoric acid, citric acid, tartaric acid,
acetic acid and trifluoroacetic acid.
[0073] Another aspect of this invention is directed to a method of
treating a protein kinase related disorder comprising a step of
administering to a patient in need thereof a therapeutically
effective amount of the compound of formula (I):
##STR00006##
its tautomer, enantiomer, diastereomer, racemate, or
pharmaceutically acceptable salt thereof, and metabolite, precursor
or prodrug thereof, or a pharmaceutically acceptable salt thereof,
or a pharmaceutical composition containing the same, wherein
R.sup.1 to R.sup.8, and X were defined as above. The method of
treating protein kinase related disorder, wherein said protein
kinase related disorder is selected from the group consisting of
the disorder related to VEGFR, EGFR, HER-2, HER-3, HER-4, PDGFR,
c-Kit, c-Met, FGFR, Ret, Flt1 and Flt3.
[0074] The method of treating protein kinase related disorder,
wherein said protein kinase related disorder is selected from the
group consisting of the disorder related to VEGFR, PDGFR, c-Kit,
Ret, Flt1 and Flt3.
[0075] The method of treating protein kinase related disorder,
wherein said protein kinase related disorder is selected from the
group consisting of leukemia, diabetes, autoimmune diseases,
hyperplasias, psoriasis, osteoarthritis, rheumatoid arthritis,
angiogenesis, cardiovascular diseases, multiple angioblastoma,
inflammatory diseases and fibrosis.
[0076] The method of treating protein kinase related disorder,
wherein said protein kinase related disorder is cancer selected
from squamous cell carcinoma, renal cell carcinoma, Kaposi's
sarcoma, non-small cell lung cancer, small cell lung cancer,
lymphoma, thyroid adenocarcinoma, breast cancer, head and neck
cancer, uterine cancer, esophageal cancer, melanoma, bladder
cancer, carcinosarcoma in urinary and genital system,
gastrointestinal carcinoma, gliomas, colorectal cancer and ovarian
cancer.
[0077] The method of treating protein kinase related disorder,
wherein said pharmaceutical composition comprising an effective
amount of a compound of formula (I) or a pharmaceutically
acceptable salt thereof and a pharmaceutically acceptable
carrier.
[0078] The method of treating protein kinase related disorder,
wherein said pharmaceutically acceptable salt is salt formed with
the acid selected from the group consisting of malic acid, lactic
acid, maleic acid, hydrochloric acid, methanesulfonic acid,
sulfuric acid, phosphoric acid, citric acid, tartaric acid, acetic
acid and trifluoroacetic acid.
[0079] The method of inhibiting the catalytic activity of a protein
kinase or the method of treating protein kinase related disorder,
wherein said compound of formula (I) comprises, but not limited to
the following:
TABLE-US-00001 Example No. Structure Name 1 ##STR00007##
(Z)-5-(2-diethylamino-ethyl)-2-(5-fluoro-2-
oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-
methyl-3a,5,6,7,8,8a-hexahydro-1H-pyrrolo [3,2-c]azepin-4-one 2
##STR00008## (Z)-2-(5-chloro-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-5-(2-diethylamino-
ethyl)-3-methyl-5,6,7,8-tetrahydro-1H- pyrrolo[3,2-c]azepin-4-one 3
##STR00009## (Z)-5-(2-diethylamino-ethyl)-2-[5-fluoro-6-
(4-fluoro-benzylamino)-2-oxo-1,2- dihydro-indol-3-ylidenemethyl]-3-
methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c] azepin-4-one 4
##STR00010## (Z)-2-(7-bromo-5-fluoro-2-oxo-1,2-
dihydro-indol-3-ylidenemethyl)-5-(2-
diethylamino-ethyl)-3-methyl-5,6,7,8- tetrahydro-1H-pyrrolo[3,2-c]
azepin-4-one 5 ##STR00011##
(Z)-2-(5-bromo-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-5-(2-diethylamino-
ethyl)-3-methyl-5,6,7,8-tetrahydro-1H- pyrrolo[3,2-c]azepin-4-one 6
##STR00012## (Z)-5-(2-diethylamino-ethyl)-2-[4-(2,3-
difluoro-phenyl)-2-oxo-1,2-dihydro-
indol-3-ylidenemethyl]-3-methyl-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 7 ##STR00013##
(Z)-N-{3-[5-(2-diethylamino-ethyl)-3-
methyl-4-oxo-1,4,5,6,7,8-hexahydro-
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-
fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl}-2- methoxy-acetamide 8
##STR00014## (S,Z)-N-{3-[5-(2-diethylamino-ethyl)-3-
methyl-4-oxo-1,4,5,6,7,8-hexahydro-
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-
fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl}-2- hydroxy-propionamide 9
##STR00015## (Z)-N-{3-[5-(2-diethylamino-ethyl)-3-
methyl-4-oxo-1,4,5,6,7,8-hexahydro-
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-
fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl}-2-
hydroxy-2-methyl-propionamide 10 ##STR00016##
(Z)-2-(5-chloro-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-3-methyl-5-(2-morpholin-4-
yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2- c]azepin-4-one 11
##STR00017## (Z)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-3-methyl-5-(2-morpholin-4-
yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2- c]azepin-4-one 12
##STR00018## (Z)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1,2-
dihydro-indol-3-ylidenemethyl]-3-methyl-5-
(2-morpholin-4-yl-ethyl)-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 13 ##STR00019##
(Z)-2-(4-bromo-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-5-(2-diethylamino-ethyl)-3-
methyl-5,6,7,8-tetrahydro-1H-pyrrolo [3,2-c]azepin-4-one 14
##STR00020## (Z)-2-(5-bromo-2-oxo-1,2-dihydro-
pyrrolo[2,3-b]pyridin-3-ylidenemethyl)-5-
(2-diethylamino-ethyl)-3-methyl-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 15 ##STR00021##
(Z)-5-(2-diethylamino-ethyl)-2-(6-
methoxy-2-oxo-1,2-dihydro-indol-3- ylidenemethyl)-3-methyl-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 16 ##STR00022##
(Z)-5-(2-diethylamino-ethyl)-3-methyl-2-
(4-methyl-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-5,6,7,8-tetrahydro-1H- pyrrolo[3,2-c]azepin-4-one 17
##STR00023## (Z)-5-(2-diethylamino-ethyl)-2-[4-(2-
hydroxy-ethyl)-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl]-3-methyl-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 18 ##STR00024##
(Z)-N-{5-fluoro-3-[3-methyl-5-(2-
morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-
hexahydro-pyrrolo[3,2-c]azepin-2-
ylmethylene]-2-oxo-2,3-dihydro-1H- indol-6-yl}-2-methoxy-acetamide
19 ##STR00025## (Z)-2-[5-fluoro-6-(4-fluoro-benzylamino)-2-
oxo-1,2-dihydro-indol-3- ylidenemethyl]-3-methyl-5-(2-
morpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-
1H-pyrrolo[3,2-c]azepin-4-one 20 ##STR00026##
(Z)-N-{5-fluoro-3-[3-methyl-5-(2-
morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-
hexahydro-pyrrolo[3,2-c]azepin-2-
ylmethylene]-2-oxo-2,3-dihydro-1H- indol-7-yl}-formamide 21
##STR00027## (S,Z)-N-{5-fluoro-3-[3-methyl-5-(2-
morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-
hexahydro-pyrrolo[3,2-c]azepin-2-
ylmethylene]-2-oxo-2,3-dihydro-1H-
indol-6-yl}-2-hydroxy-propionamide 22 ##STR00028##
(Z)-2-(5-bromo-2-oxo-1,2-dihydro-
pyrrolo[2,3-b]pyridin-3-ylidenemethyl)-3-
methyl-5-(2-morpholin-4-yl-ethyl)-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 23 ##STR00029##
(Z)-2-(5-chloro-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-5-(2-dimethylamino-
ethyl)-3-methyl-5,6,7,8-tetrahydro-1H- pyrrolo[3,2-c]azepin-4-one
24 ##STR00030## (Z)-2-(5-bromo-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-5-(2-dimethylamino-
ethyl)-3-methyl-5,6,7,8-tetrahydro-1H- pyrrolo[3,2-c]azepin-4-one
25 ##STR00031## (Z)-5-(2-dimethylamino-ethyl)-2-(5-
fluoro-2-oxo-1,2-dihydro-indol-3- ylidenemethyl)-3-methyl-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 26 ##STR00032##
(Z)-N-{3-[5-(2-dimethylamino-ethyl)-3-
methyl-4-oxo-1,4,5,6,7,8-hexahydro-
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-
fluoro-2-oxo-2,3-dihydro-1H-indol-7-yl}- formamide 27 ##STR00033##
(Z)-N-{3-[5-(2-dimethylamino-ethyl)-3-
methyl-4-oxo-1,4,5,6,7,8-hexahydro-
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-
fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl}-2- hydroxy-acetamide 28
##STR00034## (Z)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1,2-
dihydro-indol-3-ylidenemethyl]-3-methyl-5-
(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 29 ##STR00035##
(Z)-N-{5-fluoro-3-[3-methyl-4-oxo-5-(2-
pyrrolidin-1-ylethyl)-1,4,5,6,7,8-
hexahydro-pyrrolo[3,2-c]azepin-2-
ylmethylene]-2-oxo-2,3-dihydro-1H- indol-7-yl}-formamide 30
##STR00036## (Z)-N-{3-[3-methyl-4-oxo-5-(2-pyrrolidin-
1-ylethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3,
2-c]azepin-2-ylmethylene]-2-oxo-2,3-
dihydro-1H-indol-5-yl}-acetamide 31 ##STR00037##
(Z)-N-{5-fluoro-3-[3-methyl-4-oxo-5-(2-
pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-
hexahydro-pyrrolo[3,2-c]azepin-2-
ylmethylene]-2-oxo-2,3-dihydro-1H-indol-6- yl}-2-hydroxy-acetamide
32 ##STR00038## (Z)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-3-methyl-5-(2-piperidin-1-yl-
ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo [3,2-c]azepin-4-one 33
##STR00039## (Z)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1,2-
dihydro-indol-3-ylidenemethyl]-3-
methyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 34 ##STR00040##
(Z)-2-[5-(4-methoxy-phenyl)-2-oxo-1,2-
dihydro-indol-3-ylidenemethyl]-3-methyl-5-
(2-piperidin-1-yl-ethyl)-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 35 ##STR00041##
(Z)-2-(5-chloro-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-3-methyl-5-(2-piperidin-1-
yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo [3,2-c]azepin-4-one 36
##STR00042## (Z)-2-(5-bromo-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-3-methyl-5-(2-piperidin-1-
yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2- c]azepin-4-one 37
##STR00043## (Z)-N-{5-fluoro-3-[3-methyl-4-oxo-5-(2-
piperidin-1-yl-ethyl)-1,4,5,6,7,8-
hexahydro-pyrrolo[3,2-c]azepin-2-
ylmethylene]-2-oxo-2,3-dihydro-1H- indol-6-yl}-2-methoxy-acetamide
38 ##STR00044## (S,Z)-N-{5-fluoro-3-[3-methyl-4-oxo-5-(2-
piperidin-1-yl-ethyl)-1,4,5,6,7,8-
hexahydro-pyrrolo[3,2-c]azepin-2-
ylmethylene]-2-oxo-2,3-dihydro-1H-
indol-6-yl}-2-hydroxy-propionamide 39 ##STR00045##
(Z)-N-{3-[5-(2-dimethylamino-ethyl)-3-
methyl-4-oxo-1,4,5,6,7,8-hexahydro-py-
rrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-
2-oxo-2,3-dihydro-1H-indol-6-yl}-2-methoxy- acetamide 40
##STR00046## (Z)-2-[4-(2,6-difluoro-phenyl)-2-oxo-1,2-
dihydro-indol-3-ylidenemethyl]-5-(2-
dimethylamino-ethyl)-3-methyl-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 41 ##STR00047##
(Z)-5-(2-dimethylamino-ethyl)-2-[4-(3-
fluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl]-3-methyl-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 42 ##STR00048##
(Z)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1,2-
dihydro-indol-3-ylidenemethyl]-5-(2-
dimethylamino-ethyl)-3-methyl-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 43 ##STR00049##
(Z)-5-(2-dimethylamino-ethyl)-2-[5-
fluoro-6-(4-fluoro-benzylamino)-2-oxo-
1,2-dihydro-indol-3-ylidenemethyl]-3-
methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c] azepin-4-one 44
##STR00050## (Z)-N-{3-[5-(2-dimethylamino-ethyl)-3-
methyl-4-oxo-1,4,5,6,7,8-hexahydro-
pyrrolo[3,2-c]azepin-2-ylmethylene]-2-
oxo-2,3-dihydro-1H-indol-5-yl}-2- hydroxy-acetamide 45 ##STR00051##
(Z)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-3-methyl-5-(2-pyrrolidin-1-
yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo [3,2-c]azepin-4-one 46
##STR00052## (Z)-2-(5-bromo-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-3-methyl-5-(2-
pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-
pyrrolo[3,2-c]azepin-4-one 47 ##STR00053##
(Z)-2-(5-chloro-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-3-methyl-5-(2-pyrrolidin-1-
yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo [3,2-c]azepin-4-one 48
##STR00054## (Z)-3-methyl-2-(2-oxo-5-phenyl-1,2-
dihydro-indol-3-ylidenemethyl)-5-(2-
pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-
1H-pyrrolo[3,2-c]azepin-4-one 49 ##STR00055##
(Z)-2-(4-bromo-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-3-methyl-5-(2-
pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-
1H-pyrrolo[3,2-c]azepin-4-one 50 ##STR00056##
(Z)-2-(7-bromo-5-fluoro-2-oxo-1,2-
dihydro-indol-3-ylidenemethyl)-3-methyl-5-
(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-
1H-pyrrolo[3,2-c]azepin-4-one 51 ##STR00057##
(Z)-N-{3-[5-(2-diethylamino-ethyl)-3-
methyl-4-oxo-1,4,5,6,7,8-hexahydro-
pyrrolo[3,2-c]azepin-2-ylmethylene]-2-
oxo-2,3-dihydro-1H-indol-5-yl}- acetamide 52 ##STR00058##
(Z)-N-{5-fluoro-3-[3-methyl-4-oxo-5-(2-
pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-
hexahydro-pyrrolo[3,2-c]azepin-2-
ylmethylene]-2-oxo-2,3-dihydro-1H- indol-6-yl}-2-methoxy-acetamide
53 ##STR00059## (R,Z)-2-(5-fluoro-2-oxo-1,2-dihydro-
indol-3-ylidenemethyl)-5-(2-hydroxy-3-
morpholin-4-yl-propyl)-3-methyl-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 54 ##STR00060##
(Z)-3-methyl-2-(2-oxo-4-pyridin-4-yl-1,2-
dihydro-indol-3-ylidenemethyl)-5-(2-
piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-
1H-pyrrolo[3,2-c]azepin-4-one 55 ##STR00061##
(Z)-2-[5-fluoro-6-(4-fluoro-benzylamino)-2-
oxo-1,2-dihydro-indol-3-ylidenemethyl]-3-
methyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4- one
56 ##STR00062## (Z)-2-(7-bromo-5-fluoro-2-oxo-1,2-
dihydro-indol-3-ylidenemethyl)-3-methyl-5-
(2-piperidin-1-yl-ethyl)-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 57 ##STR00063##
(Z)-2-(4-bromo-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-3-methyl-5-(2-piperidin-1-
yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2- c]azepin-4-one 58
##STR00064## (Z)-3-methyl-2-(4-methyl-2-oxo-1,2-
dihydro-indol-3-ylidenemethyl)-5-(2-
piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-
pyrrolo[3,2-c]azepin-4-one 59 ##STR00065##
(R,Z)-2-(5-bromo-2-oxo-1,2-dihydro-
indol-3-ylidenemethyl)-5-(2-hydroxy-3-
morpholin-4-yl-propyl)-3-methyl-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 60 ##STR00066##
(Z)-5-(2-diethylamino-ethyl)-2-(5-fluoro-2-
oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-
methyl-6,7,8,9-tetrahydro-1H,5H-1,5-diaza-
cyclopentacycloocten-4-one 61 ##STR00067##
(Z)-2-(5-bromo-2-oxo-1,2-dihydro-indol-3-
ylidenemethyl)-5-(2-diethylamino-ethyl)-3-
methyl-6,7,8,9-tetrahydro-1H,5H-1,5-diaza-
cyclopentacycloocten-4-one 62 ##STR00068##
(Z)-5-(2-ethylamino-ethyl)-2-(5-fluoro-2-
oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-
methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c] azepin-4-one 63
##STR00069## (Z)-5-(2-diethylamino-ethyl)-2-(5-fluoro-2-
oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-
methyl-3a,5,6,7,8,8a-hexahydro-1H-pyrrolo [3,2-c]azepin-4-one
malate 64 ##STR00070## (Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-
oxoindolin-3-ylidene)methyl)-3-methyl-5-(2-
morpholinoethyl)-5,6,7,8-tetrahydropyrrolo [3,2-c]azepin-4(1H)-one
65 ##STR00071## (Z)-2-[5-(4-fluoro-phenylmethanesulfonyl)-
2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-3-
methyl-5-(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-
tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 66 ##STR00072##
(Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-
oxoindolin-3-ylidene)methyl)-3-methyl-5-(2-
(pyrrolidine-1-yl)ethyl)-5,6,7,8-
tetrahydropyrrolo[3,2-c]azepin-4(1H)-one 67 ##STR00073##
(Z)-2-((5-(4-fluorobenzylsulfonyl)-2-
oxoindolin-3-ylidene)methyl)-3-methyl-5-(2-
(piperidin-1-yl)ethyl)-5,6,7,8-
tetrahydropyrrolo[3,2-c]azepin-4(1H)-one 68 ##STR00074##
(Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-
oxoindolin-3-ylidene)methyl)-3-methyl-5-(2-
(piperidin-1-yl)ethyl)-5,6,7,8-
tetrahydropyrrolo[3,2-c]azepin-4(1H)-one 69 ##STR00075##
(Z)-2-((5-(4-fluorobenzylsulfonyl)-2-
oxoindolin-3-ylidene)methyl)-3-methyl-5-(2-
morpholinoethyl)-5,6,7,8-tetrahydropyrrolo [3,2-c]azepin-4(1H)-one
70 ##STR00076## (Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-
oxoindolin-3-ylidene)methyl)-5-(2-
(diethylamino)ethyl)-3-methyl-5,6,7,8-
tetrahydropyrrolo[3,2-c]azepin-4(1H)-one 71 ##STR00077##
(Z)-2-((5-(4-fluorobenzylsulfonyl)-2-
oxoindolin-3-ylidene)methyl)-5-(2-
(diethylamino)ethyl)-3-methyl-5,6,7,8-
tetrahydropyrrolo[3,2-c]azepin-4(1H)-one 72 ##STR00078##
(R,Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-
oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-
3-morpholinopropyl)-3-methyl-5,6,7,8-
tetrahydropyrrolo[3,2-c]azepin-4(1H)-one 73 ##STR00079##
(R,Z)-2-((5-(4-fluorobenzylsulfonyl)-2-
oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-
3-morpholinopropyl)-3-methyl-5,6,7,8-
tetrahydropyrrolo[3,2-c]azepin-4(1H)-one 74 ##STR00080##
(Z)-5-(2-(diethylamino)ethyl)-2-((4-(2,3-
difluorophenyl)-5-fluoro-2-oxoindolin-3-
ylidene)methyl)-3-methyl-5,6,7,8-
tetrahydropyrrolo[3,2-c]azepin-4(1H)-one 75 ##STR00081##
2-((Z)-(4-(2,3-difluorophenyl)-5-fluoro-2-
oxoindolin-3-ylidene)methyl)-5-((R)-2-
hydroxy-3-morpholinopropyl)-3-methyl-
5,6,7,8-tetrahydropyrrolo[3,2-c]azepin- 4(1H)-one 76 ##STR00082##
(R,Z)-5-(2-hydroxy-3-morpholinopropyl)-3-
methyl-2-((4-methyl-2-oxoindolin-3-
ylidene)methyl)-5,6,7,8-tetrahydro- pyrrolo[3,2-c]azepin-4(1H)-one
77 ##STR00083## (R,Z)-5-(2-hydroxy-3-morpholinopropyl)-2-
((6-methoxy-2-oxoindolin-3-ylidene)methyl)-
3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c] azepin-4(1H)-one 78
##STR00084## (S,Z)-2-((5-fluoro-2-oxoindolin-3-ylidene)
methyl)-5-(2-hydroxy-3-morpholinopropyl)-
3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c] azepin-4(1H)-one 79
##STR00085## (S,Z)-2-((5-chloro-2-oxoindolin-3-ylidene)
methyl)-5-(2-hydroxy-3-morpholinopropyl)-
3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c] azepin-4(1H)-one 80
##STR00086## (S,Z)-2-((5-bromo-2-oxoindolin-3-ylidene)
methyl)-5-(2-hydroxy-3-morpholinopropyl)-
3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c] azepin-4(1H)-one 81
##STR00087## (S,Z)-2-((4-bromo-2-oxoindolin-3-ylidene)
methyl)-5-(2-hydroxy-3-morpholinopropyl)-
3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c] azepin-4(1H)-one 82
##STR00088## (S,Z)-2-((7-bromo-5-fluoro-2-oxoindolin-3-
ylidene)methyl)-5-(2-hydroxy-3-morpho-
linopropyl)-3-methyl-5,6,7,8-tetrahydropy-
rrolo[3,2-c]azepin-4(1H)-one 83 ##STR00089##
(S,Z)-5-(2-hydroxy-3-morpholinopropyl)-3-
methyl-2-((4-methyl-2-oxoindolin-3-
ylidene)methyl)-5,6,7,8-tetrahydropy-
rrolo[3,2-c]azepin-4(1H)-one
[0080] In another aspect, this invention is directed to a method
for preventing or treating a mammal from protein kinase related
disorders comprising administering to the mammal a therapeutically
effective amount of the pharmaceutical composition of the
invention, which comprises the compound(s) of the invention or
pharmaceutically acceptable salts thereof, as well as
pharmaceutically acceptable carriers and excipients. The protein
kinase related disorders are selected from the group consisting of
the disorders related to VEGFR-2, EGFR, HER-2, HER-3, HER-4, PDGFR,
c-Kit, c-Met, FGFR, and Flt3. The protein kinases related disorders
also are leukemias, diabetes, autoimmune diseases, hyperplasias,
psoriasis, osteoarthritis, rheumatoid arthritis, angiogenesis,
cardiovascular diseases, von Heppel-Lindau disease, inflammatory
diseases, and fibrosis. More preferably, said protein kinase
related disorders are squamous cell carcinoma, renal cell
carcinoma, Kaposi's sarcoma, non-small cell lung cancer, small cell
lung cancer, lymphoma, thyroid adenocarcinoma, breast cancer, head
and neck cancer, uterine cancer, esophageal cancer, melanoma,
bladder cancer, carcinosarcoma in urinary and genital system,
gastrointestinal carcinoma, gliomas, colorectal cancer, ovarian
cancer. Preferably, said mammal is a human.
[0081] Further, the method for treating a mammal from protein
kianse related disorders according to the present invention is
preferably the method for treating a mammal from cancer. The
present method further comprises co-administering the mammal a
therapeutically effective amount of anti-tumor agent selected from
the group consisting of taxol or carboplatin. Preferably, said
mammal is a human. In still another aspect, this invention is
directed to a use of the compounds according to the present
invention in the preparation of a medicament for the treatment of
protein kinase related disorders. The protein kinase related
disorders are selected from the group consisting of the disorders
related to VEGFR-2, EGFR, HER-2, HER-3, HER-4, PDGFR, c-Kit, c-Met,
FGFR and Flt3. Alternatively, the protein kinases related disorders
are selected from the group consisting of leukemias, diabetes,
autoimmune diseases, hyperplasia, psoriasis, osteoarthritis,
rheumatoid arthritis, angiogenesis, cardiovascular diseases, von
Heppel-Lindau disease, inflammatory diseases, and fibrosis. More
preferably, said protein kinase related disorders are cancer
selected from the group consisting of squamous cell carcinoma,
renal cell carcinoma, Kaposi's sarcoma, non-small cell lung cancer,
small cell lung cancer, lymphoma, thyroid adenocarcinoma, breast
cancer, head and neck cancer, uterine cancer, esophageal cancer,
melanoma, bladder cancer, carcinosarcoma in urinary and genital
system, gastrointestinal carcinoma, gliomas, colorectal cancer, and
ovarian cancer.
[0082] In still another aspect, this invention is directed to the
compounds which can distinguish the protein kinase activity by
contacting cells which express the protein kinase with a compound
of the invention or a pharmaceutically acceptable salt thereof, and
determining the effect to the cell.
[0083] In still another aspect, this invention is directed to the
compounds which can distinguish protein kinase activity by
contacting man-made recombinant protein kinase with a compound of
the invention or a pharmaceutically acceptable salt thereof, and
determining kinase activity by ELISA method.
DETAILED DESCRIPTION OF THE INVENTION
[0084] Unless otherwise stated, the following terms used in the
specification and claims have the meanings discussed below.
[0085] "Alkyl" refers to a saturated aliphatic hydrocarbon radical
including C.sub.1.about.C.sub.20 straight chain and branched chain
groups. Preferably, an alkyl group is a medium size alkyl having 1
to 10 carbon atoms, e.g., methyl, ethyl, propyl, 2-propyl, n-butyl,
iso-butyl, tert-butyl, pentyl, and the like. More preferably, it is
a lower alkyl having 1 to 4 carbon atoms, e.g., methyl, ethyl,
propyl, 2-propyl, n-butyl, iso-butyl, or tert-butyl, and the like.
The alkyl group may be substituted or unsubstituted. When
substituted, the preferable substituent group(s) is halo, hydroxyl,
lower alkoxyl, aryl, aryloxyl, heteroaryl, heterocyclo alkyl,
--OR.sub.9, --NR.sub.9R.sub.10, --COR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9COR.sub.10,
--SO.sub.2R.sub.9 or --NHCO.sub.2R.sub.10.
[0086] "Cycloalkyl" refers to a 3 to 8 membered all-carbon
monocyclic ring, an all-carbon 5-membered/6-membered or
6-membered/6-membered fused bicyclic ring or a multicyclic fused
ring (a "fused" ring system means that each ring in the system
shares an adjacent pair of carbon atoms with other ring in the
system) group wherein one or more rings may contain one or more
double bonds, but none of the rings has a completely conjugated
pi-electron system. Examples of cycloalkyl groups are cyclopropane,
cyclobutane, cyclopentane, cyclopentene, cyclohexane,
cyclohexadiene, adamantane, cycloheptane, cycloheptatriene, and the
like. The cycloalkyl group may be substituted or unsubstituted.
When substituted, the substituent group(s) is preferably one or
more independently selected from the group consisting of lower
alkyl, trihalo alkyl, halo, hydroxyl, lower alkoxyl, aryl
(optionally substituted with one or more groups which each
independently is halo, hydroxyl, lower alkyl or lower alkoxyl
groups), aryloxy (optionally substituted with one or more groups
which each independently is halo, hydroxyl, lower alkyl or lower
alkoxyl groups), 6-membered heteroaryl (having 1 to 3 nitrogen
atoms in the ring, the carbons in the ring being optionally
substituted with one or more groups which each independently is
halo, hydroxyl, lower alkyl or lower alkoxyl groups), 5-membered
heteroaryl (having 1 to 3 heteroatoms selected from the group
consisting of nitrogen, oxygen and sulfur, the carbon and nitrogen
atoms of the group being optionally substituted with one or more
groups which each independently is halo, hydroxyl, lower alkyl or
lower alkoxyl groups), 5- or 6-membered heterocyclo alkyl (having 1
to 3 heteroatoms selected from nitrogen, oxygen and sulfur, the
carbon and nitrogen (if present) atoms in the group being
optionally substituted with one or more groups which each
independently is halo, hydroxyl, lower alkyl or lower alkoxyl
groups), mercapto, cyano, nitro, carboxyl, alkoxycarbonyl,
--OR.sub.9, --NR.sub.9R.sub.10, --COR.sub.S,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9COR.sub.10,
--SO.sub.2R.sub.9 and --NHCO.sub.2Rio.
[0087] "Alkenyl" refers to an alkyl group as defined above having
at least 2 carbon atoms and at least one carbon-carbon double bond.
Illustrative examples of alkenyl are derived from, but not limited
to the following: ethenyl, 1-propenyl, 2-propenyl, 1-, 2-, or
3-butenyl, and the like. Said alkenyl may be optionally substituted
with the group consisting of halo, trihalomethyl, hydroxyl, nitro,
cyano, alkoxyl, alkyl, carboxyl, alkoxycarbonyl, --OR.sub.9,
--NR.sub.9R.sub.10, --COR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9COR.sub.10,
--SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10.
[0088] "Alkynyl" refers to refers to an alkyl group as defined
above having at least 2 carbon atoms and at least one carbon-carbon
triple bond. Illustrative examples of alkynyl are derived from, but
not limited to the following: ethynyl, 1-propynyl, 2-propynyl, 1-,
2-, or 3-butynyl, and the like. Said alkynyl may be optionally
substituted with the group consisting of halo, trihalomethyl,
hydroxyl, nitro, cyano, alkoxyl, alkyl, carboxyl, alkoxycarbonyl,
--OR.sub.9, --NR.sub.9R.sub.10, --COR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9COR.sub.10,
--SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10.
[0089] "Aryl" refers to groups having at least one aromatic ring,
i.e., having a completely conjugated pi-electron system, including
all-carbon cyclic aryl, heteroaryl, fused-ring polycyclic aryl.
Said aryl may be optionally substituted with the group consisting
of halo, trihalomethyl, hydroxyl, nitro, cyano, alkoxyl, alkyl,
carboxyl, alkoxycarbonyl, --OR.sub.9, --COR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9COR.sub.10,
--SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10.
[0090] "Heteroaryl" refers to an aryl having 1 to 3 ring
heteroatoms selected from the group consisting of O, S, and N as
ring atoms, the remaining ring atoms being C. Said ring is 5 or 6
membered ring. Examples, without limitation, of heteroaryl groups
are furan, thiophene, pyridine, pyrrole, N-alkyl pyrrole,
pyrimidine, pyrazine, imidazole, and the like. Said heteroaryl may
be optionally substituted with the group consisting of halo,
trihalomethyl, hydroxyl, nitro, cyano, alkoxyl, alkyl, carboxyl,
alkoxycarbonyl, --OR.sub.9, --NR.sub.9R.sub.10, --COR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9COR.sub.10,
--SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10.
[0091] "Heterocyclo alkyl" group refers to a monocyclic or fused
ring group of 5 to 9 ring atoms having in the ring(s) one or more
atoms selected from the group consisting of nitrogen, oxygen and
S(O)n (n is integer from 0 to 2), the remaining ring atoms being C.
The rings may also have one or more double bonds, However, the
rings may or may not have a completely conjugated pi-electron
system. Examples, without limitation, of unsubstituted heterocyclo
alkyl groups are pyrrolidine, piperidine, piperazine, morpholine,
thiomorpholine, homopiperazine, and the like. The heterocyclo alkyl
may be substituted or unsubstituted. When substituted, the
substituent group(s) is preferably one or more selected from halo,
trihalomethyl, hydroxyl, nitro, cyano, alkoxyl, alkyl, carboxyl,
alkoxycarbonyl, --OR.sub.9, --NR.sub.9R.sub.10, --COR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9COR.sub.10,
--SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10.
[0092] "Hydroxyl" refers to an --OH group.
[0093] "Alkoxyl" refers to both an --O-(alkyl) and an
--O-(unsubstituted cycloalkyl) group. Representative examples
include, but are not limited to, e.g., methoxy, ethoxy, propoxy,
butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy,
cyclohexyloxy, and the like. Said alkoxyl group may be optionally
substituted with the group consisting of halo, trihalomethyl,
hydroxyl, nitro, cyano, alkoxyl, alkyl, carboxyl, alkoxycarbonyl,
--OR.sub.9, --NR.sub.9R.sub.10, --COR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9COR.sub.10,
--SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10.
[0094] "Haloalkoxy" refers to an --O-(haloalkyl). Representative
examples include, but are not limited to, trifluoromethoxy,
tribromoethoxy, and the like.
[0095] "Aryloxyl" refers to both an --O-aryl and an --O-heteroaryl
group, as defined above. Representative examples include, but are
not limited to, phenoxy, pyridinyloxy, furanyloxy, thienyloxy,
pyrimidinyloxy, pyrazinyloxy, and the like, and derivatives
thereof. Said aryloxyl group may be optionally substituted with the
group consisting of halo, trihalomethyl, hydroxyl, nitro, cyano,
alkoxyl, alkyl, carboxyl, alkoxycarbonyl, --OR.sub.9,
--NR.sub.9R.sub.10, --COR.sub.9,
--O[CH.sub.2CH.sub.2O].sub.rR.sub.11, --NR.sub.9COR.sub.13,
--SO.sub.2R.sub.9 and --NHCO.sub.2R.sub.10.
[0096] "Hydroxyalkyl" refers to a --(CH.sub.2).sub.nOH group.
[0097] "Halo" refers to fluoro, chloro, bromo, or iodo, preferably
fluoro or chloro.
[0098] "Trihalomethyl" refers to a --CX.sub.3, with X as defined
above.
[0099] "Optional" or "optionally" means that the subsequently
described event or circumstance may or may not occur, and that the
description includes instances where the event or circumstance may
or may not occur. For example, "heterocycle group optionally
substituted with an alkyl group" means that the alkyl may or may
not be present, and the description includes situations where the
heterocycle group is substituted with an alkyl group and situations
where the heterocyclo group is not substituted with the alkyl
group.
[0100] A "pharmaceutical composition" refers to a mixture of one or
more of the compounds described herein, or
physiologically/pharmaceutically acceptable salts or prodrugs
thereof, with other chemical components, such as
physiologically/pharmaceutically acceptable carriers and
excipients. The purpose of a pharmaceutical composition is to
facilitate administration of a compound to an organism.
SYNTHESIS METHOD OF THE INVENTION COMPOUND
[0101] In order to complete the object of the invention, the
invention applies the following technical solution:
[0102] The compounds of the invention can be prepared by methods
known in the art. The suitable synthetic methods are provided in
the following examples. Generally, the compounds can be prepared
according to the following scheme:
##STR00090## ##STR00091##
[0103] Reacting starting material pyrrole methyl carboxylic diester
IC-1 in tetrahydrofuran in the presence of acetic acid with
ammonium ceric nitrate at room temperature to obtain pyrrole
aldehyde carboxylic diester IC-2; reacting pyrrole aldehyde
carboxylic diester IC-2 in anhydrous tetrahydrofuran with
(carbethoxy methylene)triphenylphosphorane via Witting reaction to
obtain pyrrole ethoxycarbonyl ethenyl dicarboxylic ester IC-3;
reducing pyrrole ethoxycarbonyl ethenyl dicarboxylic ester IC-3 in
anhydrous ethanol by hydrogen catalyzed by palladium/carbon at room
temperature to obtain pyrrole ethoxycarbony ethyl dicarboxylic
ester IC-4; hydrolyzing pyrrole ethoxycarbony ethyl dicarboxylic
ester IC-4 in aqueous lithium hydroxide solution to obtain pyrrole
carboxylethyl dicarboxylic ester IC-5; reducing pyrrole
carboxylethyl dicarboxylic ester IC-5 in anhydrous tetrahydrofuran
by borane-tetrahydrofuran solution at -20.about.-5.degree. C. to
obtain pyrrole hydroxypropyl dicarboxylic ester IC-6; furthermore,
mesylating of pyrrole hydroxypropyl dicarboxylic ester IC-6 in
anhydrous dichloromethane in the presence of triethylamine at
-20.about.-5.degree. C. to obtain pyrrole methylsulfonyloxy-propyl
dicarboxylic ester IC-7; reacing pyrrole methylsulfonyloxy-propyl
dicarboxylic ester IC-7 with different amines to obtain pyrrole
amide dicarboxylic ester IC-8; reacting pyrrole amide dicarboxylic
ester IC-8 with trimethyl aluminum in toluene under reflux to
obtain the pyrrolofused seven-membered aza-heterocyclic ester IC-9;
reacting pyrrolofused seven-membered aza-heterocyclic ester IC-9
with trifluoroacetic acid at 30.about.50.degree. C. under an argon
atmosphere to obtain pyrrolofused seven-membered aza-heterocyclic
formaldehyde IC; reacting pyrrolofused seven-membered
aza-heterocyclic formaldehyde IC with indolinones in the presence
of a base such as triethylamine or piperidine under reflux for
2.about.12 hours to obtain pyrrolofused seven-membered
aza-heterocyclic derivatives (IA).
##STR00092## ##STR00093##
[0104] Reacting pyrrole aldehyde carboxylic diester IC-2 was with
Grignard reagent cyclopropylmagnesium bromide in anhydrous
tetrahydrofuran at room temperature under an argon atmosphere to
obtain pyrrole cyclopropyl hydroxycarboxylic diester ID-1; reacting
pyrrole cyclopropyl hydroxycarboxylic diester ID-1 with hydrobromic
acid in methanol to obtain bromo-butenyl pyrrole diester ID-2;
reducing bromo-butenyl pyrrole diester ID-2 in anhydrous ethanol by
hydrogen catalyzed by palladium/carbon at room temperature to
obtain bromo-butyl pyrrole diester ID-3; reacting bromo-butyl
pyrrole diester ID-3 with different amines in dichloromethane under
reflux to obtain pyrrole amide dicarboxylic diester ID-4; reacting
pyrrole amide dicarboxylic diester ID-4 with trimethyl aluminum in
toluene under reflux to obtain the pyrrolofused eight-membered
aza-heterocyclic aldehyde ID; reacting pyrrolofused eight-membered
aza-heterocyclic aldehyde ID with indolinones in the presence of a
base such as triethylamine or piperidine under reflux for 2-12
hours to obtain pyrrolofused eight-membered aza-heterocyclic
derivatives (IB).
[0105] Wherein, the double bond of Formula (I) was Z
configuration(cis), which can be confirmed by the NMR data.
Generally, the chemical shift of pyrrole NH proton was about 9 ppm,
but that of the pyrrole NH proton of the obtained compounds was
about 14 ppm. The downfield shift of NH proton was mainly because
of intramolecular hydrogen bonding interaction between pyrrole NH
proton and adjacent carbonyl oxygen atom of oxindole, which was
also described in patent WO0160814(Su-11248).
[0106] This invention also relates to a pharmaceutical composition
comprising compounds or salts thereof of this invention in an
effective therapeutic dose, as well as pharmaceutically acceptable
carrier.
[0107] Furthermore, this invention relates to a use of the
compounds of formula (I) or salts thereof in the preparation of a
medicament as tyrosine kinase inhibitors. In other words, this
invention also provides the composition comprising the above
compound in an effective therapeutic dose, and the use of the
compounds and/or pharmaceutical compositions containing them in the
preparation of a medicament as tyrosine kinase inhibitors.
[0108] The following examples serve to illustrate the invention,
but the examples should not be considered as limiting the scope of
the invention.
EXAMPLES
[0109] The structures of the compounds were confirmed by nuclear
magnetic resonance (NMR) or mass spectrometry (MS). NMR shifts
(.delta.) were given in parts per million (ppm). NMR measurements
were performed on a Bruker AVANCE-400, using deutorated chloroform
(CDCl.sub.3) and deutorated dimethylsulfoxide (DMSO-D.sub.6) as
solvents and Tetramethylsilane (TMS) as an internal reference,
chemical shifts are given in parts per million (ppm).
[0110] MS measurements were performed on a FINNIGAN LCQAd (ESI)
mass spectrometer.
[0111] The average of inhibitory rate of kinase VEGFR was
determined by the HTScan (Cell Signaling).
[0112] The average of inhibitory rate of kinase EGFR/HER-2 was
determined by the NovoStar (BMG LABTECH in Germany).
[0113] Thin-layer silica gel was Yantai Huanghai HSGF254 or Qingdao
GF254 silica gel plates.
[0114] Column chromatography generally used Yantai Huanghai 200-300
mesh silica gel as carrier.
[0115] DMSO-D.sub.6: deutorated dimethyl sulfoxide.
[0116] CDCl.sub.3: deutorated chloroform.
PREPARATION EXAMPLES
Example 1
(Z)-5-(2-Diethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidene-
methyl)-3-methyl-3a,5,6,7,8,8a-hexahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00094##
##STR00095## ##STR00096##
[0117] Step 1
5-Formyl-3-methyl-1H-pyrrole-2,4-dicarboxylic acid 2-tert-butyl
ester 4-ethyl ester
[0118] 3,5-Dimethyl-1H-pyrrole-2,4-dicarboxylic acid 2-tert-butyl
ester 4-ethyl ester 1a (30 g, 0.113 mol) was dissolved in 300 ml of
tetrahydrofuran, and added with 360 ml of acetic acid and 300 ml of
water under stirring at room temperature. Upon completion of the
addition, the mixture was stirred to mix well, and added with
ammonium ceric nitrate (246 g, 0.449 mol) in one portion. The
mixture was stirred at room temperature for 0.5 hour, and the color
of the reaction solution turned from nacarat to orange. After thin
lay chromatography showed the disappearance of starting materials,
the reaction mixture was poured into 800 ml of ice water, and light
yellow precipitates were formed. The mixture was stirred for
another 0.5 hour, filtered and dried in vacuo to obtain the title
compound 5-formyl-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester 1b (31.13 g, yield 98%) as a light
yellow solid.
[0119] MS m/z (ESI): 282.0[M+1]
Step 2
5-(2-Ethoxycarbonyl-vinyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester
[0120] 5-Formyl-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester 1b (23 g, 81.7 mmol) and
(ethoxycarbonylmethylene)triphenylphosphorane (34.66 g, 99.4 mmol)
were dissolved in 450 ml of tetrahydrofuran, and stirred at room
temperature overnight under an argon atmosphere. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was concentrated under reduced pressure to obtain
a yellow oil. The residue was dissolved in the solvent mixture of
n-hexane and ethyl acetate (V:V=20:1), and purified by sand funnel
decompression column chromatography to obtain the title compound
5-(2-ethoxycarbonyl-vinyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester 1c (24 g, yield 84%) as a
light yellow solid.
[0121] MS m/z (ESI): 352.1[M+1]
Step 3
5-(2-Ethoxycarbonyl-ethyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester
[0122]
5-(2-Ethoxycarbonyl-vinyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester 1c (24 g, 68.3 mmol) was
dissolved in anhydrous ethanol (180 ml) under stirring, and added
with palladium on activated carbon (2.44 g, 10%) to the solution.
The resulting solution was stirred at room temperature overnight
under a hydrogen atmosphere. After thin lay chromatography showed
the disappearance of starting materials, the reaction mixture was
filtered to remove palladium on activated carbon, and washed with a
little ethanol. The filtrate was concentrated under reduced
pressure to obtain the title compound
5-(2-ethoxycarbonyl-ethyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester 1d (23 g, yield 95%) as a
white solid.
[0123] MS m/z (ESI): 354.40 [M+1]
Step 4
5-(2-Carboxy-ethyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester
[0124]
5-(2-Ethoxycarbonyl-ethyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester 1d (23.6 g, 66.8 mmol) was
dissolved in 190 ml of tetrahydrofuran and 90 ml of methanol under
stirring, and added with aqueous lithium hydroxide solution (80 ml,
10 mol/L, 0.8 mol) at room temperature. The color of the reaction
solution gradually turned from light yellow to cyan, and stirred
for another 1 hour. After thin lay chromatography showed the
disappearance of starting materials, the resulting mixture was
concentrated under reduced pressure to evaporate organic solvent.
The residue was adjusted pH to 2 with hydrochloric acid solution (2
mol/L) in an ice-water bath under stirring. White precipitates were
formed. The mixture was filtered, and the filter cake was dried to
obtain the title compound
5-(2-carboxy-ethyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester le (24 g, yield 98%) as a white
solid.
[0125] MS m/z (ESI): 326.1[M+1]
Step 5
5-(3-Hydroxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester
[0126] 5-(2-Carboxy-ethyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester le (9.75 g, 30 mmol) was
dissolved in 90 ml of anhydrous tetrahydrofuran under stirring, and
added ropwise slowly with a solution of borane in tetrahydrofuran
(90 ml, 1 mol/L, 90 mmol) to the solution while maintaining the
temperature at -10.about.-5.degree. C. in an ice-salt bath under an
argon atmosphere. Upon completion of the addition, the ice-salt
bath was removed and the reaction mixture was allowed to warm up to
room temperature and stirred for 2-3 hours. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was concentrated under reduced pressure to
evaporate the solvent. The residue was added with 100 ml of
saturated sodium bicarbonate solution and 100 ml of ethyl acetate,
and stirred until dissolved. The resulting mixture was extracted
with ethyl acetate (100 ml.times.3). The combined organic extracts
were washed with 100 ml of saturated brine, dried over anhydrous
magnesium sulfate, filtered to remove the drying agent and
concentrated under reduced pressure to obtain the title compound
5-(3-hydroxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester if (9.2 g, yield 98%) as a light
yellow oil.
[0127] MS m/z (ESI): 312.3[M+1]
Step 6
5-(3-Methanesulfonyloxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester
[0128] 5-(3-Hydroxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester if (9.20 g, 30 mmol) was
dissolved in 150 ml of dichloromethane under stirring, and added
with triethylamine (7.0 ml, 50 mmol) to the solution while
maintaining the temperature at about -10.degree. C. in an ice-salt
bath under an argon atmosphere. Upon completion of the addition,
the mixture was added slowly with methanesulfonyl chloride (3.5 ml,
45 mmol). After stirring to mix well, the reaction system was
allowed to warm up to room temperature and stirred for 4 hours.
After thin lay chromatography showed the disappearance of starting
materials, the reaction mixture was quenched with ice. The reaction
mixture was washed successively with dilute hydrochloric acid (0.5
mol/L, 80 ml.times.2) to remove triethylamine, saturated sodium
carbonate solution (80 ml.times.2) to remove excess hydrochloric
acid and saturated brine (80 ml), and concentrated under reduced
pressure to obtain the title compound
5-(3-methanesulfonyloxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester 1g (11.4 g, yield 99%) as a
brown oil.
[0129] MS m/z (ESI): 390.5[M+1]
Step 7
5-[3-(2-Diethylamino-ethylamino)-propyl]-3-methyl-1H-pyrrole-2,4-dicarboxy-
lic acid 2-tert-butyl ester 4-ethyl ester
[0130]
5-(3-Methanesulfonyloxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxyli-
c acid 2-tert-butyl ester 4-ethyl ester 1g (8.24 g, 21 mmol) was
dissolved in N,N-diethylethylenediamine (15 ml, 100 mmol) under
stirring at room temperature, and stirred overnight. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was added with 100 ml of ethyl acetate and 100 ml
of saturated brine, stirred for 5 minutes, and separated into
layers. The organic phase was washed with saturated brine (100
ml.times.4), dried over anhydrous magnesium sulfate, filtered to
remove the drying agent and concentrated under reduced pressure to
obtain brown oil. The residue was purified by silica gel column
chromatography to obtain the title compound
5-[3-(2-diethylamino-ethylamino)-propyl]-3-methyl-1H-pyrrole-2,4-dicarbox-
ylic acid 2-tert-butyl ester 4-ethyl ester 1 h (8.2 g, yield 95%)
as a colorless oil.
[0131] MS m/z (ESI): 410.2[M+1]
Step 8
5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2--
c]azepine-2-carboxylic acid tert-butyl ester
[0132]
5-[3-(2-Diethylamino-ethylamino)-propyl]-3-methyl-1H-pyrrole-2,4-di-
carboxylic acid 2-tert-butyl ester 4-ethyl ester 1 h (3.547 g, 8.67
mmol) was dissolved in 70 ml of toluene and stirred for 10 minutes
at room temperature under an argon atmosphere. The mixture was
added with a solution of trimethyl aluminum in toluene (5.6 ml, 2
mol/L, 11.27 mmol), and stirred for another 30 minutes at room
temperature until no white smoke was released. The reaction mixture
was heated to reflux for 4 hours in an oil bath. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was naturally cooled down to room temperature,
quenched with ethanol (10 ml, 95%), and added with anhydrous
ethanol (60 ml). The resulting mixture was filtered through a pad
of Celite, washed with anhydrous ethanol (200 ml.times.4) and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography to obtain the title compound
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carboxylic acid tert-butyl ester 1i (0.413 g, yield
75.7%) as a white solid.
[0133] MS m/z (ESI): 364.1[M+1]
Step 9
5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2--
c]azepine-2-carbaldehyde
[0134]
5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrro-
lo[3,2-c]azepine-2-carboxylic acid tert-butyl ester 1i (0.413 g,
1.14 mmol) was dissolved in trifluoroacetic acid (1.5 ml, 20 mmol)
under stirring, heated to 40.degree. C. for 5 minutes in an oil
bath under an argon atmosphere, and cooled down to -5.degree. C. in
an ice-salt bath under stirring, and added with triethoxy methane
(0.34 ml, 1.7 mmol) and stirred for 2 minutes. Then the ice-salt
bath was removed, and the reaction mixture was allowed to warm up
to room temperature and stirred for another about 2 hours. After
thin lay chromatography showed the disappearance of starting
materials, the reaction mixture was added with 3 ml of ice water
and 10 ml of dichloromethane, adjusted to pH 11 with aqueous sodium
hydroxide solution (2 mol/L) and extracted with dichloromethane (10
ml.times.3). The combined organic extracts were dried over
anhydrous magnesium sulfate, filtered to remove the drying agent
and concentrated under reduced pressure to obtain yellow oil. The
residue was purified by silica gel column chromatography to obtain
the title compound
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j (0.271 g, yield 55%) as a light brown
oil.
[0135] MS m/z (ESI): 292.3[M+1]
Step 10
(Z)-5-(2-Diethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidene-
methyl)-3-methyl-3a,5,6,7,8,8a-hexahydro-1H-pyrrolo[3,2-c]azepin-4-one
[0136]
5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrro-
lo[3,2-c]azepine-2-carbaldehyde 1j (0.271 g, 0.93 mmol) and
5-fluoro-1,3-dihydro-indol-2-one (0.127 g, 0.84 mmol) were
dissolved in 1.4 ml of anhydrous ethanol under stirring at room
temperature, the resulting mixture was stirred for 10 minutes in
dark, and added with piperidine (0.15 ml, 1.49 mmol). The mixture
was refluxed at 70.degree. C. for about 1.5 hours in an oil bath
under an argon atmosphere and lots of orange precipitates were
formed. After thin lay chromatography showed the disappearance of
starting materials, the ice-salt bath was removed, and the reaction
mixture was naturally cooled down to room temperature, filtered and
dried to obtain the title compound
(Z)-5-(2-diethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-yliden-
emethyl)-3-methyl-3a,5,6,7,8,8a-hexahydro-1H-pyrrolo[3,2-c]azepin-4-one
1(0.288 g, yield 80.76%) as an orange solid.
[0137] MS m/z (ESI): 425.3[M+1]
[0138] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.710 (s, 1H,
pyrrole-NH), 10.903 (s, 1H, indole-NH), 7.753.about.7.782 (dd, 1H,
--ArH), 7.744 (s, 1H, --CH.dbd.C), 6.914.about.6.965 (m, 1H,
--ArH), 6.834.about.6.867 (m, 1H, --ArH), 3.483-3.518 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.336-3.364 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.907.about.2.944 (t, 2H,
seven-membered ring intra-CH.sub.2C.dbd.C), 2.529.about.2.581 (m,
6H, 3.times.-CH.sub.2N), 2.455 (s, 3H, pyrrole-CH.sub.3),
2.040-2.079 (m, 2H, seven-membered ring intra --CH.sub.2),
0.956.about.0.992 (t, 6H, 2.times.-CH.sub.3).
Example 2
(Z)-2-((5-Chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-diethylamin-
o-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00097##
[0140] The title compound was prepared under the same conditions as
described in step 10 of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example 1 and
5-chloro-1,3-dihydro-indol-2-one as starting materials to obtain
(Z)-2-((5-chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-diethylami-
no-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
2 (27 mg, yield 60.0%) as a nacarat solid.
[0141] MS m/z (ESI): 441.1[M+1]
[0142] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.663 (s, 1H,
pyrrole-NH), 11.002 (s, 1H, indole-NH), 7.987-7.991 (d, 1H, --ArH),
7.798 (s, 1H, --CH.dbd.C), 7.132-7.158 (dd, 1H, --ArH), 6.867-6.888
(d, 1H, --ArH), 3.483-3.518 (t, 2H, seven-membered ring
intra-CH.sub.2N), 3.336.about.3.364 (t, 2H, amide N seven-membered
ring outer-CH.sub.2), 2.907.about.2.944 (t, 2H, seven-membered ring
intra-CH.sub.2C.dbd.C), 2.529-2.581 (m, 6H, 3.times.-CH.sub.2N),
2.455 (s, 3H, pyrrole-CH.sub.3), 2.040-2.079 (m, 2H, seven-membered
ring intra-CH2), 0.956.about.0.992 (1, 6H, 2.times.-CH.sub.3).
Example 3
(Z)-5-(2-Diethylamino-ethyl)-2-[5-fluoro-6-(4-fluoro-benzylamino)-2-oxo-1,-
2-dihydro-indol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,-
2-c]azepin-4-one
##STR00098##
##STR00099##
[0143] Step 1
(5-Fluoro-2,4-dinitro-phenyl)-acetic acid
[0144] (3-Fluoro-phenyl)-acetic acid 3a (31.5 g, 0.204 mol) was
dissolved in sulfuric acid (64 ml, 98%) under stirring at room
temperature, and added dropwise with the mixture (V:V=1:1, 100 ml)
of nitric acid (65%-68%) and sulfuric acid (98%) while maintaining
the temperature at about 35.degree. C. Upon completion of the
addition, the mixture was stirred at 35.degree. C. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was added with ice and filtered after ice-out to
obtain the title compound (5-fluoro-2,4-dinitro-phenyl)-acetic acid
3b (49 mg) as a light yellow oil.
[0145] MS m/z (ESI): 243.5[M-1]
Step 2
(2,4-Diamino-5-fluoro-phenyl)-acetic acid
[0146] (5-Fluoro-2,4-dinitro-phenyl)-acetic acid 3b (10 g, 38.7
mmol) was dissolved in 150 ml of methanol under stirring, and added
with palladium on activated carbon (1.5 g, 5%) to the solution at
room temperature. The reaction mixture was hydrogenated under 0.3
Mpa of hydrogen. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
filtered twice, and concentrated under reduced pressure to obtain
the title compound (2,4-diamino-5-fluoro-phenyl)-acetic acid 3c
(7.12 g) as a brown solid to the next step.
Step 3
5-Fluoro-6-amino-indol-2-one
[0147] (2,4-Diamino-5-fluoro-phenyl)-acetic acid 3c (7.12 g, 38.7
mmol) was dissolved in hydrochloric acid (100 ml, 1 mol/L) under
stirring at room temperature. The solution was heated to reflux for
1 hour. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was cooled down to room
temperature, neutralized with sodium hydroxide solution (100 ml, 1
mol/L) in an ice-water bath, extracted with ethyl acetate (125
ml.times.4). The combined organic extracts were washed with
saturated brine (100 ml), dried over anhydrous magnesium sulfate,
filtered to remove the drying agent and concentrated under reduced
pressure to obtain the title compound 5-fluoro-6-amino-indol-2-one
3d (5.3 g, yield 82.8%) as a yellow solid.
[0148] MS m/z (ESI): 165.3[M-1]
Step 4
5-Fluoro-6-(4-fluoro-benzylamino)-1,3-dihydro-indol-2-one
[0149] 6-Amino-5-fluoro-1,3-dihydro-indol-2-one 3d (2.26 g, 13.6
mmol) was dissolved in 40 ml of ethanol under stirring at room
temperature, the resulting solution was cooled down to 0.degree. C.
in an ice-water bath, and added with 4-fluoro-benzaldehyde (1.5 ml,
13.6 mmol) to the solution. Upon completion of the addition, the
resulting solution was stirred for 1 hour at room temperature,
added with sodium borohydride (1.08 g, 28.5 mmol) and heated to
reflux for 18 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature, added with ice water and
lots of precipitates were formed, filtered and washed with water
(50 ml.times.3). The residue was purified by silica gel column
chromatography to obtain the title compound
5-fluoro-6-(4-fluoro-benzylamino)-1,3-dihydro-indol-2-one 3e (1.67
g, yield 45%) as a white solid.
[0150] MS m/z (ESI): 275[M+1]
Step 5
(Z)-5-(2-Diethylamino-ethyl)-2-[5-fluoro-6-(4-fluoro-benzylamino)-2-oxo-1,-
2-dihydro-indol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,-
2-c]azepin
[0151] The title compound was prepared under the same conditions as
described in step of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example 1 and
5-fluoro-6-(4-fluoro-benzylamino)-1,3-dihydro-indol-2-one 3e as
starting materials to obtain
(Z)-5-(2-diethylamino-ethyl)-2-[5-fluoro-6-(4-fluoro-benzylamino)-2-oxo-1-
,2-dihydro-indol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3-
,2-c]azepin-4-one 3 (61 mg, yield 62.2%) as a henna solid.
[0152] MS m/z (ESI): 548.3[M+1]
[0153] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.416 (s, 1H,
pyrrole-NH), 10.520 (s, 1H, indole-NH), 7.573-7.602 (d, 1H, --ArH),
7.366-7.401 (m, 2H, --ArH), 7.350 (s, 1H, --CH.dbd.C),
7.141.about.7.185 (m, 2H, --ArH), 6.410.about.6.415 (m, 1H, --ArH),
6.038.about.6.057 (d, 1H, --ArH), 4.346.about.4.361 (d, 2H,
aniline-CH.sub.2), 3.466.about.3.501 (t, 2H, seven-membered ring
intra-CH.sub.2N), 3.336.about.3.364 (t, 2H, amide N seven-membered
ring outer-CH.sub.2), 2.907.about.2.944 (t, 2H, seven-membered ring
intra-CH.sub.2C.dbd.C), 2.529.about.2.581 (m, 6H,
3.times.-CH.sub.2N), 2.388 (s, 3H, pyrrole-CH.sub.3),
2.011.about.2.039 (m, 2H, seven-membered ring intra-CH.sub.2),
0.967.about.1.063 (t, 6H, 2.times.-CH.sub.3).
Example 4
(Z)-2-(7-Bromo-5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-diet-
hylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00100##
##STR00101##
[0154] Step 1
7-Bromo-5-fluoro-1,3-dihydro-indol-2-one
[0155] 5-Fluoro-1,3-dihydro-indol-2-one 4a (1.5 g, 0.01 mol) was
dissolved in 15 ml of acetonitrile under stirring, and added
dropwise with N-bromosuccinimide (1.8 g, 0.01 mol) at room
temperature. Upon completion of the addition, the mixture was
stirred overnight and lots of precipitate was formed. After thin
lay chromatography showed the disappearance of starting materials,
the reaction mixture was filtered to obtain
7-bromo-5-fluoro-1,3-dihydro-indol-2-one 4b (2 g, yield 87%) as a
gray solid.
[0156] MS m/z (ESI): 228.3[M-1]
Step 2
(Z)-2-(7-Bromo-5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-diet-
hylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
[0157] The title compound was prepared under the same conditions as
described in step of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example 1 and
7-bromo-5-fluoro-1,3-dihydro-indol-2-one 4b as starting materials
to obtain
(Z)-2-(7-bromo-5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-
-(2-diethylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepi-
n-4-one 4 (55 mg, yield 61.1%) as a yellow solid.
[0158] MS m/z (ESI): 503.6[M+1]
[0159] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.653 (s, 1H,
pyrrole-NH), 11.181 (s, 1H, indole-NH), 7.848.about.7.876 (dd, 1H,
--ArH), 7.794 (s, 1H, --CH.dbd.C), 7.242.about.7.270 (dd, 1H,
--ArH), 3.485.about.3.520 (t, 2H, seven-membered ring
intra-CH.sub.2N), 3.338.about.3.366 (t, 2H, amide N seven-membered
ring outer-CH.sub.2), 2.932.about.2.969 (t, 2H, seven-membered ring
intra-CH.sub.2C.dbd.C), 2.527.about.2.582 (m, 6H,
3.times.-CH.sub.2N), 2.470 (s, 3H, pyrrole-CH.sub.3),
2.031.about.2.093 (m, 2H, seven-membered ring intra-CH.sub.2),
0.954.about.0.990 (t, 6H, 2.times.-CH.sub.3).
Example 5
(Z)-2-((5-Bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-diethylamino-
-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00102##
[0161] The title compound was prepared under the same conditions as
described in step of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example and
5-bromo-1,3-dihydro-indol-2-one as starting materials to obtain
(Z)-2-((5-bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-diethylamin-
o-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
5 (59 mg, yield 67.8%) as a yellow solid.
[0162] MS m/z (ESI): 485.5 [M+1]
[0163] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.660 (s, 1H,
pyrrole-NH), 11.008 (s, 1H, indole-NH), 8.113.about.8.117 (d, 1H,
--ArH), 7.803 (s, 1H, --CH.dbd.C), 7.260.about.7.286 (dd, 1H,
--ArH), 6.825.about.6.845 (d, 1H, --ArH), 3.482.about.3.516 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.336.about.3.364 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 2.907.about.2.944 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.490.about.2.578
(m, 6H, 3.times.-CH.sub.2N), 2.464 (s, 3H, pyrrole-CH.sub.3),
2.039-2.067 (m, 2H, seven-membered ring intra-CH.sub.2),
0.954.about.0.990 (t, 6H, 2.times.-CH.sub.3).
Example 6
(Z)-5-(2-Diethylamino-ethyl)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1,2-dihydro--
indol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-
-4-one
##STR00103##
##STR00104##
[0164] Step 1
4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indole
[0165] 4-Bromo-1H-indole 6a (29.4 g, 150 mmol) was dissolved in 600
ml of dimethyl sulfoxide under stirring, and added successively
with bis(pinacolato)diboron (41.9 g, 165 mmol), potassium acetate
(44.1 g, 450 mmol) and
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (3.6 g, 4.8
mmol) under an argon atmosphere. Upon completion of the addition,
the reaction mixture was stirred at 80.degree. C. in an oil bath
for 22 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was added
with water (2 L) and extracted with ethyl acetate (2 L.times.3).
The combined organic extracts were washed with saturated brine (2
L.times.5), dried over anhydrous sodium sulfate, filtered to remove
the drying agent and concentrated under reduced pressure. The
residue was purified by silica gel column chromatography and
recrystallized to obtain
4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indole 6b (20
g, yield 60%) as a white solid.
[0166] MS m/z (ESI): 243.9[M+1]
Step 2
4-(2,3-Difluoro-phenyl)-1H-indole
[0167] 4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indole
6b (1.22 g, 5 mmol) was dissolved in 20 ml of tetrahydrofuran under
stirring, and added with 1-bromo-2,3-difluoro-benzene (0.97 g, 5
mmol), tetrakis (triphenylphosphine)palladium (0.17 g, 0.15 mmol)
and sodium hydroxide solution (7 ml, 2 mol/L) under an argon
atmosphere. Upon completion of the addition, the reaction system
was stirred at 75.degree. C. in an oil bath overnight. After thin
lay chromatography showed the disappearance of starting materials,
the reaction mixture was naturally cooled down to room temperature
and extracted with ethyl acetate (20 ml.times.3). The combined
organic extracts were washed with saturated brine (10 ml), dried
over anhydrous sodium sulfate, filtered to remove the drying agent
and concentrated under reduced pressure. The residue was purified
by silica gel column chromatography to obtain
4-(2,3-difluoro-phenyl)-1H-indole 6c (800 mg, yield 70%) as a white
solid.
[0168] MS m/z (ESI): 228.4[M-1]
Step 3
4-(2,3-Difluoro-phenyl)-1,3-dihydro-indol-2-one
[0169] 4-(2,3-difluoro-phenyl)-1H-indole 6c (744 mg, 3.25 mmol) was
dissolved in 12 ml of ethanol under stirring, and added
successively with tert-butanol (21 ml), glacial acetic acid (6.4
ml) and pyridinium tribromide (3.12 g, 9.7 mmol) at room
temperature. Upon completion of the addition, the reaction mixture
was stirred for 3 hours, added with glacial acetic acid (16 ml) and
zinc dust (1.1 g, 16.25 mmol), and stirred for another 1 hour. The
reaction mixture was filtered and concentrated under reduced
pressure. The residue was added with ethyl acetate (30 ml), washed
successively with water (10 ml), saturated sodium bicarbonate
solution (10 ml) and saturated brine (10 ml), dried over anhydrous
sodium sulfate, filtered to remove the drying agent and
concentrated under reduced pressure to obtain
4-(2,3-difluoro-phenyl)-1,3-dihydro-indol-2-one 6d (780 mg, yield
97%) as a white solid.
[0170] MS m/z (ESI): 246.6[M+1]
Step 4
(Z)-5-(2-Diethylamino-ethyl)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1,2-dihydro--
indol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-
-4-one
[0171] The title compound was prepared under the same conditions as
described in step 10 of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example 1 and
4-(2,3-difluoro-phenyl)-1,3-dihydro-indol-2-one 6d as starting
materials to obtain
(Z)-5-(2-diethylamino-ethyl)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1-
,2-dihydro-indol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3-
,2-c]azepin-4-one 6 (43 mg, yield 61.4%) as a yellow solid.
[0172] MS m/z (ESI): 519.6[M+1]
[0173] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.531 (s, 1H,
pyrrole-NH), 11.133 (s, 1H, indole-NH), 7.617.about.7.640 (m, 1H,
--ArH), 7.429.about.7.442 (m, 1H, --ArH), 7.305.about.7.340 (m, 1H,
--ArH), 7.232.about.7.270 (m, 1H, --ArH), 6.997.about.7.017 (d, 1H,
--ArH), 6.874.about.6.893 (d, 1H, --ArH), 6.710 (s, 1H,
--CH.dbd.C), 3.445.about.3.478 (t, 2H, seven-membered ring
intra-CH.sub.2N), 3.313 (m, 2H, amide N seven-membered ring
outer-CH.sub.2), 2.868-3.904 (t, 2H, seven-membered ring
intra-CH.sub.2C.dbd.C), 2.465.about.2.542 (m, 6H,
3.times.-CH.sub.2N), 2.002.about.2.032 (m, 2H, seven-membered ring
intra-CH.sub.2), 1.794 (s, 3H, pyrrole-CH.sub.3), 0.930-0.965 (t,
6H, 2.times.-CH.sub.3).
Example 7
(Z)--N-{3-[5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-p-
yrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-y-
l}-2-methoxy-acetamide
##STR00105##
##STR00106##
[0174] Step 1
N-(5-Fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-methoxy-acetamide
[0175] 6-Amino-5-fluoro-1,3-dihydro-indol-2-one 3d (2.028 g, 12.2
mmol) was dissolved in 30 ml of tetrahydrofuran under stirring, and
added with 1.3 ml of pyridine to the solution at room temperature.
The reaction system was cooled down to about -50.degree. C. in a
dry ice-ethanol bath. A solution of methoxy-acetyl chloride (1.35
g, 12.5 mmol) in tetrahydrofuran (20 ml) was added dropwise to the
above reaction system. Upon completion of the addition, the
ice-ethanol bath was removed, and the reaction mixture was allowed
to warm up to room temperature and stirred overnight. After thin
lay chromatography showed the disappearance of starting materials,
the reaction mixture was filtered. The resulting solid was washed
with water (10 mL.times.3) and recrystallized with methanol to
obtain
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-methoxy-acetamide 7a
(1.18 mg, yield 40.6%) as a gray solid.
[0176] MS m/z (ESI): 239.3[M+1]
Step 2
(Z)--N-{3-[5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-p-
yrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-y-
l}-2-methoxy-acetamide
[0177] The title compound was prepared under the same conditions as
described in step of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example 1 and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-methoxy-acetamide 7a
as starting materials to obtain
(Z)--N-{3-[5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro--
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6--
yl}-2-methoxy-acetamide 7 (37 mg, yield 53.6%) as a brown
solid.
[0178] MS m/z (ESI): 512.5[M+1]
[0179] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.605 (s, 1H,
pyrrole-NH), 10.893 (s, 1H, indole-NH), 9.320 (s, 1H, amide-NH),
7.838.about.7.866 (d, 1H, --ArH), 7.350 (s, 1H, --CH.dbd.C),
7.540.about.7.556 (d, 1H, --ArH), 4.064 (s, 2H, --CH.sub.2O),
3.483.about.3.517 (t, 2H, seven-membered ring intra-CH.sub.2N),
3.336.about.3.362 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 3.314 (s, 3H, --CH.sub.3O), 2.902.about.2.939 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.530.about.2.562
(m, 6H, 3.times.-CH.sub.2N), 2.444 (s, 3H, pyrrole-CH.sub.3),
2.037.about.2.066 (m, 2H, seven-membered ring intra-CH.sub.2),
0.958.about.0.993 (t, 6H, 2.times.-CH.sub.3).
Example 8
(S,Z)--N-{3-[5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-
-pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-
-yl}-2-hydroxy-propionamide
##STR00107##
##STR00108##
[0180] Step 1
Acetic acid
1-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-ylcarbamoyl)-ethyl
ester
[0181] 5-Fluoro-6-amino-1,3-dihydro-indol-2-one 3d (450 mg, 2.71
mmol) was dissolved in 10 ml of tetrahydrofuran under stirring at
room temperature. The mixture was cooled down to -45.degree. C. in
a dry ice-acetone bath and added with 364 .mu.l of piperidine. A
solution of acetic acid 1-chlorocarbonyl-ethyl ester (423 mg, 2.71
mmol) in 10 ml of tetrahydrofuran was added dropwise to the above
reaction system. Upon completion of the addition, the ice-acetone
bath was removed, and the reaction mixture was allowed to warm up
to room temperature and stirred overnight. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was filtered. The filter cake was washed with
water, and the resulting solid was dried to obtain the title
compound acetic acid
1-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-ylcarbamoyl)- ethyl ester
8a (840 mg) as a white solid to be used directly in the next
step.
[0182] MS m/z (ESI): 281.5[M+1]
Step 2
N-(5-Fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-propionamide
[0183] Acetic acid
1-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-ylcarbamoyl)-ethyl ester
8a (1.86 g, 6.4 mmol) was dissolved in 20 ml of methanol under
stirring, and added with 10 ml of water and sodium hydroxide
solution (10 ml, 0.7 mol/L) to the solution and stirred for 4 hours
at room temperature. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
neutralized with hydrochloride acid (1 mol/L) and concentrated
under reduced pressure. The residue was purified by silica gel
column chromatography and dried to obtain
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-propionamide
8b (1.0 g, yield 70%) as a white solid.
[0184] MS m/z (ESI): 239.6 [M+1]
Step 3
(S,Z)--N-{3-[5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-
-pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-
-yl}-2-hydroxy-propionamide
[0185] The title compound was prepared under the same conditions as
described in step 10 of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example 1 and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-propionamide
8b as starting materials to obtain
(S,Z)--N-{3-[5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydr-
o-pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol--
6-yl}-2-hydroxy-propionamide 8 (28 mg, yield 40.8%) as a yellow
solid.
[0186] MS m/z (ESI): 512.4[M+1]
[0187] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.594 (s, 1H,
pyrrole-NH), 10.902 (s, 1H, indole-NH), 9.245 (s, 1H, amide-NH),
7.856.about.7.884 (d, 1H, --ArH), 7.725.about.7.741 (d, 1H, --ArH),
7.663 (s, 1H, --CH.dbd.C), 6.057.about.6.070 (d, 1H, --ArH),
4.206.about.4.236 (q, 1H, --CHO), 3.480.about.3.514 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.336.about.3.362 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 2.902.about.2.939 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.530.about.2.562
(m, 6H, 3.times.-CH.sub.2N), 2.443 (s, 3H, pyrrole-CH.sub.3),
2.037.about.2.066 (m, 2H, seven-membered ring intra-CH.sub.2),
1.328.about.1.345 (d, 3H, --CH.sub.3), 0.958.about.0.993 (t, 6H,
2.times.-CH.sub.3).
Example 9
(Z)--N-{3-[5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-p-
yrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-y-
l}-2-hydroxy-2-methyl-propionamide
##STR00109##
##STR00110##
[0188] Step 1
Acetic acid
1-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-ylcarbamoyl)-1-methyl-ethyl
ester
[0189] 5-Fluoro-6-amino-1,3-dihydro-indol-2-one 3d (410 mg, 2.47
mmol) was dissolved in 10 ml of tetrahydrofuran under stirring at
room temperature. The mixture was cooled down to -45.degree. C. in
a dry ice-acetone bath and added with piperidine (322 .mu.l). A
solution of acetic acid 1-chlorocarbonyl-1-methyl-ethyl ester (423
mg, 2.71 mmol) in tetrahydrofuran (10 ml) was added dropwise to the
above reaction system. Upon completion of the addition, the
ice-acetone bath was removed, and the reaction mixture was allowed
to warm up to room temperature and stirred overnight. After thin
lay chromatography showed the disappearance of starting materials,
the reaction mixture was filtered. The filter cake was washed with
water, and the resulting solid was dried to obtain acetic acid
1-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-ylcarbamoyl)-1-methyl-ethyl
ester 9a (792 mg) as a white solid to the next step.
[0190] MS m/z (ESI): 293.7[M-1]
Step 2
N-(5-Fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-2-methyl-propionami-
de
[0191] Acetic acid
1-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-ylcarbamoyl)-ethyl ester
9a (2.035 g, 6.9 mmol) was dissolved in 20 ml of methanol under
stirring, added with sodium hydroxide solution (20 ml, 0.7 mol/L)
(20 ml) to the solution and stirred for 4 hours at room
temperature. After thin lay chromatography showed the disappearance
of starting materials, the reaction mixture was neutralized with
hydrochloride acid (1 mol/L) and concentrated under reduced
pressure. The residue was purified by silica gel column
chromatography and dried to obtain
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-2-methyl-propionam-
ide 9b (900 mg, yield 59.2%) as a white solid.
[0192] MS m/z (ESI): 253.6[M+1]
Step 3
(Z)--N-{3-[5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-p-
yrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-y-
l}-2-hydroxy-2-methyl-propionamide
[0193] The title compound was prepared under the same conditions as
described in step 10 of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-2-methyl-propionam-
ide 9b as starting materials to obtain
(Z)--N-{3-[5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro--
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6--
yl}-2-hydroxy-2-methyl-propionamide 9 (39 mg, yield 62.4%) as a
yellow solid.
[0194] MS m/z (ESI): 526.4 [M+1]
[0195] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.591 (s, 1H,
pyrrole-NH), 10.900 (s, 1H, indole --NH), 9.284 (s, 1H, amide-NH),
7.862.about.7.890 (d, 1H, --ArH), 7.774.about.7.791 (d, 1H, --ArH),
7.661 (s, 1H, --CH.dbd.C), 6.052 (s, 1H, --OH), 3.480.about.3.514
(t, 2H, seven-membered ring intra-CH.sub.2N), 3.334.about.3.361 (t,
2H, amide N seven-membered ring outer-CH.sub.2), 2.902.about.2.939
(t, 2H, seven-membered ring intra-CH.sub.2C.dbd.C),
2.530.about.2.562 (m, 6H, 3.times.-CH.sub.2N), 2.443 (s, 3H,
pyrrole-CH.sub.3), 2.037.about.2.066 (m, 2H, seven-membered ring,
intra-CH.sub.2), 1.377 (s, 6H, 2.times.-CH.sub.3), 0.958-0.993 (t,
6H, 2.times.-CH.sub.3).
Example 10
(Z)-2-((5-Chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-mo-
rpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00111##
##STR00112##
[0196] Step 1
3-Methyl-5-[3-(2-morpholin-4-yl-ethylamino)-propyl]-1H-pyrrole-2,4-dicarbo-
xylic acid 2-tert-butyl ester 4-ethyl ester
[0197]
5-(3-Methanesulfonyloxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxyli-
c acid 2-tert-butyl ester 4-ethyl ester 1g (5.812 g, 15 mmol)
obtained from step 6 of Example 1 and 2-morpholin-4-yl-ethylamine
(10.725 g, 82.5 mmol) was dissolved in water bath at 30.degree. C.
and stirred for 5.5 hours at room temperature. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was added with ethyl acetate (100 ml) and
saturated brine (100 ml), stirred for 5 minutes, and separated into
layers. The organic phase was washed with saturated brine (100
ml.times.4), dried over anhydrous magnesium sulfate, filtered to
remove the drying agent and concentrated under reduced pressure.
The residue was purified by silica gel column chromatography to
obtain
3-methyl-5-[3-(2-morpholin-4-yl-ethylamino)-propyl]-1H-pyrrole-2,4-
-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester 10a (2.238 g,
yield 87%) as a light yellow oil.
[0198] MS m/z (ESI): 424.9[M+1]
Step 2
3-Methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carboxylic acid tert-butyl ester
[0199]
3-Methyl-5-[3-(2-morpholin-4-yl-ethylamino)-propyl]-1H-pyrrole-2,4--
dicarboxylic acid 2-tert-butyl ester 4-ethyl ester 10a (2.238 g,
5.29 mmol) was dissolved in 50 ml of toluene under stirring, and
added slowly with a solution of trimethyl aluminum in toluene (3.9
ml, 2 mol/L, 7.9 mmol) under an argon atmosphere. The reaction
system was stirred for 30 minutes at room temperature until no
white smoke was released, and refluxed for another 3 hours in an
oil bath. After thin lay chromatography showed the disappearance of
starting materials, the oil bath was removed, and the reaction
mixture was quenched with a little water, adjusted to pH 8-10 with
dilute sodium hydroxide solution (2 mol/L), added with saturated
brine (50 ml) and extracted with ethyl acetate (50 ml.times.3). The
combined organic extracts were filtered through a pad of Celite,
concentrated under reduced pressure to obtain the title compound
3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carboxylic acid tert-butyl ester 10b (1.218 g, yield
61%) as a light yellow solid.
[0200] MS m/z (ESI): 378.2[M+1]
Step 3
3-Methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde
[0201]
3-Methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyr-
rolo[3,2-c]azepine-2-carboxylic acid tert-butyl ester 10b (725 mg,
1.92 mmol) was dissolved in trifluoroacetic acid (2.6 ml, 34.2
mmol) in an ice-water bath under stirring. The reaction mixture was
stirred at 40.degree. C. in a water bath for 5 minutes, added with
triethoxy methane (0.42 ml, 2.5 mmol) in one portion at -5.degree.
C. in an ice-water bath, and stirred for 2 minutes. Then the
ice-salt bath was removed, and the reaction mixture was allowed to
warm up to room temperature, became brown and stirred for another 2
hours. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was quenched with a little
water, adjusted to pH 8 with dilute sodium hydroxide solution (2
mol/L) and extracted with dichloromethane (50 ml.times.3). The
combined organic extracts were concentrated under reduced pressure
to obtain henna solid. The solid was purified by silica gel column
chromatography to obtain the title compound
3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 10c (240 mg, yield 40%) as a light
yellow solid.
[0202] MS m/z (ESI): 306.3[M+1]
Step 4
(Z)-2-((5-Chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-mo-
rpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
[0203]
3-Methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyr-
rolo[3,2-c]azepine-2-carbaldehyde 10c (53 mg, 0.174 mmol) and
5-chloro-1,3-dihydro-indol-2-one (29 g, 0.84 mmol) was dissolved in
0.9 ml of ethanol under stirring, and added with piperidine (0.1
ml, 1.0 mmol) to the solution at room temperature. The mixture was
heated to reflux for 2 hours in an oil bath and lots of
precipitates were formed. Then the ice-salt bath was removed, and
the reaction mixture was naturally cooled down to room temperature,
filtered to obtain the title compound
(Z)-2-((5-chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-meth-
yl-5-(2-morpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-
-one 10 (30 g, yield 38%) as a red solid.
[0204] MS m/z (ESI): 455.2[M+1]
[0205] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.682 (s, 1H,
pyrrole-NH), 11.009 (s, 1H, indole-NH), 7.992.about.7.997 (d, 1H,
--ArH), 7.804 (s, 1H, --CH.dbd.C), 7.138.about.7.164 (dd, 1H,
--ArH), 6.873.about.6.894 (d, 1H, --ArH), 3.572.about.3.583 (m, 6H,
N seven-membered ring --CH.sub.2, 2.times.-CH.sub.2O),
3.346.about.3.360 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 2.938.about.2.974 (t, 2H, --CH.sub.2C.dbd.C),
2.463 (s, 3H, pyrrole-CH.sub.3), 2.438.about.2.510 (m, 6H,
2.times.-CH.sub.2N), 2.054.about.2.083 (m, 2H, seven-membered ring
intra-CH.sub.2).
Example 11
(Z)-2-((5-Fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-mo-
rpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00113##
[0207] The title compound was prepared under the same conditions as
described in step 4 of Example 10 with
3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 10c obtained from step 3 of Example 10
and 5-fluoro-1,3-dihydro-indol-2-one as starting materials to
obtain
(Z)-2-((5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-m-
orpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
11 (29 mg, yield 51.5%) as a yellow solid.
[0208] MS m/z (ESI): 439.3[M+1]
[0209] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.727 (s, 1H,
pyrrole-NH), 10.908 (s, 1H, indole-NH), 7.756.about.7.785 (dd, 1H,
--ArH), 7.746 (s, 1H, --CH.dbd.C), 6.917.about.6.968 (m, 1H,
--ArH), 6.838.about.6.871 (m, 1H, --ArH), 3.571.about.3.600 (m, 6H,
N seven-membered ring-CH.sub.2, 2.times.-CH.sub.2O),
3.344.about.3.371 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 2.934.about.2.971 (t, 2H, --CH.sub.2C.dbd.C),
2.454 (s, 3H, pyrrole-CH.sub.3), 2.438.about.2.510 (m, 6H,
2.times.-CH.sub.2N), 2.05.about.2.082 (m, 2H, seven-membered ring
intra-CH.sub.2).
Example 12
(Z)-2-[4-(2,3-Difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-3--
methyl-5-(2-morpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azep-
in-4-one
##STR00114##
[0211] The title compound was prepared under the same conditions as
described in step 4 of Example 10 with
3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 10c obtained from step 3 of Example 10
and 4-(2,3-difluoro-phenyl)-1,3-dihydro-indol-2-one 6d obtained
from step 3 of Example 6 as starting materials to obtain
(Z)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-3-
-methyl-5-(2-morpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]aze-
pin-4-one 12 (20 mg, yield 29%) as a yellow solid.
[0212] MS m/z (ESI): 533.3 [M+1]
[0213] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.546 (s, 1H,
pyrrole-NH), 11.135 (s, 1H, indole-NH), 7.598.about.7.662 (m, 1H,
--ArH), 7.409.about.7.461 (m, 1H, --ArH), 7.306.about.7.343 (m, 1H,
--ArH), 7.234.about.7.273 (m, 1H, --ArH), 6.700.about.7.019 (d, 1H,
--ArH), 6.875.about.6.894 (d, 1H, --ArH), 6.712 (s, 1H,
--CH.dbd.C), 3.547.about.3.560 (m, 6H, N seven-membered
ring-CH.sub.2, 2.times.-CH.sub.2O), 3.295.about.3.310 (t, 2H, amide
N seven-membered ring outer-CH.sub.2), 2.897.about.2.933 (t, 2H,
--CH.sub.2C.dbd.C), 2.410.about.2.510 (m, 6H, 2.times.-CH.sub.2N),
1.999.about.2.061 (m, 2H, seven-membered ring intra-CH.sub.2),
1.794 (s, 3H, pyrrole-CH.sub.3).
Example 13
(Z)-2-(4-Bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-diethylamino--
ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00115##
[0215] The title compound was prepared under the same conditions as
described in step of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example 1 and
4-bromo-1,3-dihydro-indol-2-one as starting materials to obtain
(Z)-2-(4-bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-diethylamino-
-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
13 (30 mg, yield 45.5%) as an orange solid.
[0216] MS m/z (ESI): 585.1[M+1]
[0217] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.634 (s, 1H,
pyrrole-NH), 11.184 (s, 1H, indole-NH), 8.589 (s, 1H, --CH.dbd.C),
7.220.about.7.240 (d, 1H, --ArH), 7.056.about.7.096 (m, 1H, --ArH),
6.938.about.6.957 (d, 1H, --ArH), 3.491.about.3.524 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.346.about.3.373 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 2.930.about.2.966 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.508.about.2.569
(m, 6H, 3.times.-CH.sub.2N), 2.412 (s, 3H, pyrrole-CH.sub.3),
2.030.about.2.095 (m, 2H, seven-membered ring intra-CH.sub.2),
0.959.about.0.994 (t, 6H, 2.times.-CH.sub.3).
Example 14
(Z)-2-((5-Bromo-2-oxo-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenemethyl)-5-
-(2-diethylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepi-
n-4-one
##STR00116##
[0219] The title compound was prepared under the same conditions as
described in step 10 of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example 1 and
5-bromo-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one as starting
materials to obtain
(Z)-2-((5-bromo-2-oxo-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenem-
ethyl)-5-(2-diethylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-
-c]azepin-4-one 14 (23 mg, yield 33.8%) as an orange solid.
[0220] MS m/z (ESI): 486.2[M+1]
[0221] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.472 (s, 1H,
pyrrole-NH), 11.636 (s, 1H, indole-NH), 8.487 (s, 1H, pyridine-CH),
8.116 (s, 1H, pyridine-CH), 7.889 (s, 1H, --CH.dbd.C), 3.505 (t,
2H, seven-membered ring intra-CH.sub.2N), 3.354 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.930.about.2.966 (t, 2H,
seven-membered ring intra-CH.sub.2C.dbd.C), 2.508.about.2.569 (m,
6H, 3.times.-CH.sub.2N), 2.472 (s, 3H, pyrrole-CH.sub.3),
2.030.about.2.095 (m, 2H, seven-membered ring intra-CH.sub.2),
0.959.about.0.994 (t, 6H, 2.times.-CH.sub.3).
Example 15
(Z)-5-(2-Diethylamino-ethyl)-2-(6-methoxy-2-oxo-1,2-dihydro-indol-3-yliden-
emethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00117##
[0223] The title compound was prepared under the same conditions as
described in step of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example and
6-methoxy-1,3-dihydro-indol-2-one as starting materials to obtain
(Z)-5-(2-diethylamino-ethyl)-2-(6-methoxy-2-oxo-1,2-dihydro-indol-3-ylide-
nemethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
15 (31 mg, yield 52.7%) as a red solid.
[0224] MS m/z (ESI): 437.4[M+1]
[0225] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.466 (s, 1H,
pyrrole-NH), 10.852 (s, 1H, indole-NH), 7.670.about.7.691 (d, 1H,
--ArH), 7.496 (s, 1H, --CH.dbd.C), 6.575.about.6.596 (d, 1H,
--ArH), 6.463 (s, 1H, --ArH), 3.769 ((s, 3H, --CH.sub.3),
3.477.about.3.510 (t, 2H, seven-membered ring intra-CH.sub.2N),
3.316.about.3.347 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 2.888.about.2.924 (t, 2H, seven-membered ring
intra-CH.sub.2C.dbd.C), 2.489.about.2.573 (m, 6H,
3.times.-CH.sub.2N), 2.416 (s, 3H, pyrrole-CH.sub.3),
2.030.about.2.095 (m, 2H, seven-membered ring intra-CH.sub.2),
0.954.about.0.982 (t, 6H, 2.times.-CH.sub.3).
Example 16
(Z)-5-(2-Diethylamino-ethyl)-3-methyl-2-(4-methyl-2-oxo-1,2-dihydro-indol--
3-ylidenemethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00118##
[0227] The title compound was prepared under the same conditions as
described in step of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example and
4-methyl-1,3-dihydro-indol-2-one as starting materials to obtain
(Z)-5-(2-diethylamino-ethyl)-3-methyl-2-(4-methyl-2-oxo-1,2-dihydro-indol-
-3-ylidenemethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
16 (25 mg, yield 44.1%) as a yellow solid.
[0228] MS m/z (ESI): 421.5[M+1]
[0229] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.710 (s, 1H,
pyrrole-NH), 10.927 (s, 1H, indole-NH), 7.566 (s, 1H, --CH.dbd.C),
7.034.about.7.072 (m, 1H, --ArH), 6.768.about.6.835 (dd, 2H,
--ArH), 3.483.about.3.517 (t, 2H, seven-membered ring
intra-CH.sub.2N), 3.339.about.3.366 (t, 2H, amide N seven-membered
ring outer-CH.sub.2), 2.909.about.2.946 (t, 2H, seven-membered ring
intra-CH.sub.2C.dbd.C), 2.489.about.2.573 (m, 6H,
3.times.-CH.sub.2N), 2.590 (s, 3H, benzene-CH.sub.3), 2.386 (s, 3H,
pyrrole-CH.sub.3), 2.041.about.2.069 (m, 2H, seven-membered ring
intra-CH.sub.2), 0.956.about.0.991 (t, 6H, 2.times.-CH.sub.3).
Example 17
(Z)-5-(2-Diethylamino-ethyl)-2-[4-(2-hydroxy-ethyl)-2-oxo-1,2-dihydro-indo-
l-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-o-
ne
##STR00119##
[0231] The title compound was prepared under the same conditions as
described in step of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example 1 and
4-(2-hydroxy-ethyl)-1,3-dihydro-indol-2-one as starting materials
to obtain
(Z)-5-(2-diethylamino-ethyl)-2-[4-(2-hydroxy-ethyl)-2-oxo-1,2-dihy-
dro-indol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]az-
epin-4-one 17 (18 mg, yield 29.5%) as a yellow solid.
[0232] MS m/z (ESI): 451.5 [M+1]
[0233] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.722 (s, 1H,
pyrrole-NH), 10.924 (s, 1H, indole-NH), 7.652 (s, 1H, --CH.dbd.C),
7.045.about.7.065 (m, 1H, --ArH), 6.772.about.6.847 (dd, 2H,
--ArH), 4.871 (s, 1H, --OH), 3.726.about.3.739 (t, 2H, amide N
seven-membered ring outer-CH.sub.2O), 3.486.about.3.502 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.342.about.3.556 (t, 2H,
--CH.sub.2), 3.095 (t, 2H, benzene-CH.sub.2), 2.910.about.2.946 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.504.about.2.566
(m, 6H, 3 x-CH.sub.2N), 2.398 (s, 3H, pyrrole-CH.sub.3), 2.092 (m,
2H, seven-membered ring intra-CH.sub.2), 0.957.about.0.993 (t, 6H,
2.times.-CH.sub.3).
Example 18
(Z)--N-{5-Fluoro-3-[3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8--
hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-6-
-yl}-2-methoxy-acetamide
##STR00120##
[0235] The title compound was prepared under the same conditions as
described in step 4 of Example 10 with
3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 10c obtained from step 3 of Example 10
and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-methoxy-acetamide 7a
obtained from step 1 of Example 7 as starting materials to obtain
(Z)--N-{5-fluoro-3-[3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-
-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol--
6-yl}-2-methoxy-acetamide 18 (47 mg, yield 60%) as a yellow
solid.
[0236] MS m/z (ESI): 526.1[M+1]
[0237] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.608 (s, 1H,
pyrrole-NH), 10.884 (s, 1H, indole-NH), 9.308 (s, 1H, --NHCO),
7.827.about.7.854 (d, 1H, --ArH), 7.659 (s, 1H, --CH.dbd.C),
7.534.about.7.550 (d, 1H, --ArH), 4.056 (s, 2H, --CH.sub.2O),
3.561.about.3.571 (m, 6H, N seven-membered ring-CH.sub.2,
2.times.-CH.sub.2O), 3.397 (s, 3H, --CH.sub.3O), 3.331.about.3.345
(t, 2H, amide N seven-membered ring outer-CH.sub.2),
2.918.about.2.954 (t, 2H, --CH.sub.2C.dbd.C), 2.432 (s, 3H,
pyrrole-CH.sub.3), 2.431.about.2.500 (m, 6H, 2.times.-CH.sub.2N),
2.040.about.2.067 (m, 2H, seven-membered ring intra-CH.sub.2).
Example 19
(Z)-2-[5-Fluoro-6-(4-fluoro-benzylamino)-2-oxo-1,2-dihydro-indol-3-ylidene-
methyl]-3-methyl-5-(2-morpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[-
3,2-c]azepin-4-one
##STR00121##
[0239] The title compound was prepared under the same conditions as
described in step 4 of Example 10 with
3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 10c obtained from step 3 of Example 10
and 5-fluoro-6-(4-fluoro-benzylamino)-1,3-dihydro-indol-2-one 3e
obtained from step 4 of Example 3 as starting materials to obtain
(Z)-2-[5-fluoro-6-(4-fluoro-benzylamino)-2-oxo-1,2-dihydro-indol-3-yliden-
emethyl]-3-methyl-5-(2-morpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo-
[3,2-c]azepin-4-one 19 (57 mg, yield 69%) as a carmine solid.
[0240] MS m/z (ESI): 526.1[M+1]
[0241] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.423 (s, 1H,
pyrrole-NH), 10.515 (s, 1H, indole-NH), 7.565.about.7.595 (d, 1H,
--ArH), 7.359.about.7.394 (m, 2H, --ArH), 7.343 (s, 1H,
--CH.dbd.C), 7.134.about.7.177 (m, 1H, --ArH), 6.404 (m, 1H, --NH),
6.032.about.3.051 (d, 1H, --ArH), 4.399.about.4.353 (d, 2H,
amide-CH.sub.2), 3.544.about.3.555 (m, 6H, N seven-membered
ring-CH.sub.2, 2.times.-CH.sub.2O), 3.310.about.3.326 (t, 2H, amide
N seven-membered ring outer-CH.sub.2), 2.870.about.2.906 (t, 2H,
--CH.sub.2C.dbd.C), 2.378 (s, 3H, pyrrole-CH.sub.3),
2.416.about.2.500 (m, 6H, 2.times.-CH.sub.2N), 2.014.about.2.041
(m, 2H, seven-membered ring intra-CH.sub.2).
Example 20
(Z)--N-{5-Fluoro-3-[3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8--
hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-7-
-yl}-formamide
##STR00122##
##STR00123##
[0242] Step 1
5-Fluoro-7-nitro-indol-2-one
[0243] 5-Fluoro-1,3-dihydro-indol-2-one 4a (5.0 g, 33 mmol) was
dissolved in sulfuric acid (17.6 ml, 98%) under stirring at
-5.degree. C., and nitric acid (2.1 ml, 65%-68%) was added to the
solution while maintaining the temperature below 0.degree. C. Upon
completion of the addition, the mixture was stirred at room
temperature for 1 hour. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
poured into ice water and filtered after ice-out. The filter cake
was washed with water for three times, and the resulting solid was
recrystallized to obtain the title compound
5-fluoro-7-nitro-1,3-dihydro-indol-2-one 20a (4.0 g, yield 62.5%)
as an orange solid.
[0244] MS m/z (ESI): 196.3[M+1]
Step 2
7-Amino-5-fluoro-1,3-dihydro-indol-2-one
[0245] 5-Fluoro-7-nitro-1,3-dihydro-indol-2-one 20a (4.0 g, 20
mmol) was dissolved in 200 ml of acetic acid under stirring, and
added with palladium on activated carbon (1.0 g, 5%) at room
temperature. The reaction system was stirred under a hydrogen
atmosphere. After thin lay chromatography showed the disappearance
of starting materials, the reaction mixture was filtered, and
concentrated under reduced pressure to obtain the title compound
7-amino-5-fluoro-1,3-dihydro-indol-2-one 20b (3.2 g, yield 97.5%)
as a white solid.
[0246] MS m/z (ESI): 167.4[M+1]
Step 3
5-Fluoro-7-formamido-indol-2-one
[0247] A mixture of acetic anhydride (0.8 ml) and formic acid (0.6
ml) was stirred for 1 hour at room temperature, and added with
7-amino-5-fluoro-1,3-dihydro-indol-2-one 20b (2.0 g, 12 mmol) in 30
ml of tetrahydrofuran to the above mixture, followed by piperidine
(0.02 ml). The resulting mixture was stirred for 3 hours until
precipitates were formed, filtered to provide the crude product
(1.95 g), and recrystallized from methanol to obtain the title
compound 5-fluoro-7-formamido-indol-2-one 20c (700 mg, yield 30.4%)
as a white solid.
[0248] MS m/z (ESI): 195.1[M+1]
Step 4
(Z)--N-{5-Fluoro-3-[3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8--
hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-7-
-yl}-formamide
[0249] The title compound was prepared under the same conditions as
described in step of Example 10 with
3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 10c obtained from step 3 of Example 10
and N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-7-yl)-formamide 20c as
starting materials to obtain
(Z)--N-{5-fluoro-3-[3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-
-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol--
7-yl}-formamide 20 (37 mg, yield 52%) as an orange solid.
[0250] MS m/z (ESI): 480.2[M-1]
[0251] .sup.1HNMR (400 MHz, DMSO-d6) .delta.13.655 (s, 1H,
pyrrole-NH), 10.424 (s, 1H, indole-NH), 9.801 (s, 1H, --NHCO),
8.330 (s, 1H, --CHO), 7.757 (s, 1H, --CH.dbd.C), 7.610.about.7.633
(d, 1H, --ArH), 7.428.about.7.461 (dd, 1H, --ArH),
3.562.about.3.592 (m, 6H, N seven-membered ring-CH.sub.2,
2.times.-CH.sub.2O), 3.331.about.3.345 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.935.about.2.971 (t, 2H,
--CH.sub.2C.dbd.C), 2.451 (s, 3H, pyrrole-CH.sub.3),
2.431.about.2.500 (m, 6H, 2.times.-CH.sub.2N), 2.046.about.2.074
(m, 2H, seven-membered ring intra-CH.sub.2).
Example 21
(S,Z)--N-{5-Fluoro-3-[3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-
-6-yl}-2-hydroxy-propionamide
##STR00124##
[0253] The title compound was prepared under the same conditions as
described in step 4 of Example 10 with
3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 10c obtained from step 3 of Example 10
and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-propionamide
8b obtained from step 2 of Example 8 as starting materials to
obtain
(S,Z)--N-{5-fluoro-3-[3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7-
,8-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indo-
l-6-yl}-2-hydroxy-propionamide 21 (44 mg, yield 58%) as an orange
solid.
[0254] MS m/z (ESI): 526.1[M+1]
[0255] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.599 (s, 1H,
pyrrole-NH), 10.895 (s, 1H, indole-NH), 9.236 (s, 1H, --NHCO),
7.846.about.7.874 (d, 1H, --ArH), 7.718.about.7.734 (d, 1H, --ArH),
7.663 (s, 1H, --CH.dbd.C), 6.051.about.6.064 (d, 1H, --OH),
4.199.about.4.229 (t, 1H, --CHO), 3.560.about.3.586 (m, 6H, N
seven-membered ring-CH.sub.2, 2.times.-CH.sub.2O),
3.331.about.3.345 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 2.918.about.2.954 (t, 2H, --CH.sub.2C.dbd.C),
2.456 (s, 3H, pyrrole-CH.sub.3), 2.431.about.2.500 (m, 6H,
2.times.-CH.sub.2N), 2.048.about.2.077 (m, 2H, seven-membered ring
intra-CH.sub.2), 1.321.about.1.338 (d, 2H, --CH.sub.3).
Example 22
(Z)-2-((5-Bromo-2-oxo-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenemethyl)-3-
-methyl-5-(2-morpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]aze-
pin-4-one
##STR00125##
[0257] The title compound was prepared under the same conditions as
described in step 4 of Example 10 with
3-methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 10c obtained from step 3 of Example 10
and 5-bromo-1,3-dihydro-pyrrolo[2,3-b]pyridin-2-one as starting
materials to obtain
(Z)-2-((5-bromo-2-oxo-1,2-dihydro-pyrrolo[2,3-b]pyridin-3-ylidenem-
ethyl)-3-methyl-5-(2-morpholin-4-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3-
,2-c]azepin-4-one 22 (59 mg, yield 60%) as an orange solid.
[0258] MS m/z (ESI): 500.0[M+1]
[0259] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.474 (s, 1H,
pyrrole-NH), 11.004 (s, 1H, indole-NH), 8.475 (d, 1H, pyridine-CH),
8.102.about.8.107 (d, 1H, pyridine-CH), 7.873 (s, 1H, --CH.dbd.C),
3.560.about.3.591 (m, 6H, N seven-membered ring-CH.sub.2,
2.times.-CH.sub.2O), 3.336.about.3.364 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.946.about.2.983 (t, 2H,
--CH.sub.2C.dbd.C), 2.456 (s, 3H, pyrrole-CH.sub.3),
2.425.about.2.500 (m, 6H, 2.times.-CH.sub.2N), 2.048.about.2.077
(m, 2H, seven-membered ring intra-CH.sub.2).
Example 23
(Z)-2-((5-Chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-dimethylami-
no-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00126##
##STR00127##
[0260] Step 1
5-[3-(2-Dimethylamino-ethylamino)-propyl]-3-methyl-1H-pyrrole-2,4-dicarbox-
ylic acid 2-tert-butyl ester 4-ethyl ester
[0261]
5-(3-Methanesulfonyloxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxyli-
c acid 2-tert-butyl ester 4-ethyl ester 1g (11.964 g, 25.7 mmol)
obtained from step 6 of Example 1 was dissolved in
N,N-dimethylethylenediamine (12 ml, 97 mmol), the resulting
solution was stirred for 5 hours at room temperature. After thin
lay chromatography showed the disappearance of starting materials,
the reaction mixture was added with ethyl acetate (80 ml) and
saturated brine (80 ml), stirred for 5 minutes, and separated into
layers. The organic phase was washed with saturated brine (80
ml.times.4) to remove N,N-dimethylethylenediamine, dried over
anhydrous magnesium sulfate, filtered to remove the drying agent
and concentrated under reduced pressure to obtain brown oil. The
residue was purified by silica gel column chromatography to obtain
the title compound
5-[3-(2-dimethylamino-ethylamino)-propyl]-3-methyl-1H-pyrrole-2,4-dicarbo-
xylic acid 2-tert-butyl ester 4-ethyl ester 23a (5.85 g, yield
45.9%) as a yellow oil.
[0262] MS m/z (ESI): 382.2 [M+1]
Step 2
5-(2-Dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carboxylic acid tert-butyl ester
[0263]
5-[3-(2-Dimethylamino-ethylamino)-propyl]-3-methyl-1H-pyrrole-2,4-d-
icarboxylic acid 2-tert-butyl ester 4-ethyl ester 23a (5.85 g, 13.8
mmol) was dissolved in 130 ml of toluene under stirring, and added
slowly with a solution of trimethyl aluminum in toluene (12 ml, 2
mol/L, 24 mmol) to the solution under an argon atmosphere. Upon the
completion of the addition, the reaction mixture was stirred for 10
minutes at room temperature until no white smoke was released, and
heated to reflux for 3 hours in an oil bath. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was quenched with ice water. After the reaction
system was naturally cooled down to room temperature, the mixture
was added with hydrochloric acid solution (50 ml, 2 mol/L) and
stirred for 10 minutes. The mixture was adjusted to pH 9 with
aqueous sodium hydroxide solution (2 mol/L), and extracted with
dichloromethane (50 ml.times.4). The combined organic extracts were
dried over anhydrous magnesium sulfate, filtered to remove the
drying agent and concentrated under reduced pressure to obtain the
title compound
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carboxylic acid tert-butyl ester 23b (3.3 g, yield
71.4%) as a yellow solid.
[0264] MS m/z (ESI): 336.2[M+1]
Step 3
5-(2-Dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde
[0265]
5-(2-Dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrr-
olo[3,2-c]azepine-2-carboxylic acid tert-butyl ester 23b (774 mg,
2.3 mmol) was dissolved in trifluoroacetic acid (3.1 ml, 20 mmol)
under stirring, the resulting solution was heated at 40.degree. C.
for 5 minutes in an oil bath under an argon atmosphere. The
reaction mixture was cooled down to -5.degree. C. in an ice-salt
bath under stirring, added with triethoxy methane (0.5 ml, 3.0
mmol) and stirred for 2 minutes. Then the ice-salt bath was
removed, and the reaction mixture was allowed to warm up to room
temperature and stirred for another 1 hour. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was added with ice water (3 ml) and
dichloromethane (10 ml), adjusted to pH 11 with aqueous sodium
hydroxide solution (2 mol/L) and extracted with dichloromethane (10
ml.times.3). The combined organic extracts were dried over
anhydrous magnesium sulfate, filtered to remove the drying agent
and concentrated under reduced pressure to obtain a henna oil. The
residue was purified by silica gel column chromatography to obtain
the title compound
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c (223 mg, yield 37%) as a yellow
oil.
[0266] MS m/z (ESI): 264.2[M+1]
Step 4
(Z)-2-((5-Chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-dimethylami-
no-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
[0267]
5-(2-Dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrr-
olo[3,2-c]azepine-2-carbaldehyde 23c (53 mg, 0.2 mmol) was
dissolved in 1 ml of methanol under stirring, and added with
5-chloro-1,3-dihydro-indol-2-one (34 mg, 0.2 mmol) and piperidine
(0.1 ml) to the solution. Upon the completion of the addition, the
mixture was stirred to mix well in dark, heated to reflux for 2
hours and lots of precipitate was formed. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was naturally cooled down to room temperature, and
filtered. The filter cake was washed with ethanol and dried to
obtain the title compound
(Z)-2-((5-chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-dimethylam-
ino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
23 (62 mg, yield 75%) as an orange powder.
[0268] MS m/z (ESI): 413.1[M+1]
[0269] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.667 (s, 1H,
pyrrole-NH), 11.004 (s, 1H, indole-NH), 7.990 (s, 1H, --ArH), 7.799
(s, 1H, --CH.dbd.C), 7.134.about.7.159 (m, 1H, --ArH),
6.869.about.6.890 (m, 1H, --ArH), 3.533.about.3.567 (t, 2H, N
seven-membered ring-CH.sub.2), 3.336.about.3.364 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.909.about.2.945 (t, 2H,
--CH.sub.2C.dbd.C), 2.463 (s, 3H, pyrrole-CH.sub.3),
2.401.about.2.434 (t, 2H, --CH.sub.2N), 2.204 (s, 6H,
2.times.-CH.sub.3N), 2.035.about.2.079 (m, 2H, seven-membered ring
intra --CH.sub.2).
Example 24
(Z)-2-((5-Bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-dimethylamin-
o-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00128##
[0271] The title compound was prepared under the same conditions as
described in step 4 of Example 23 with
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c obtained from step 3 of Example 23
and 5-bromo-1,3-dihydro-indol-2-one as starting materials to obtain
(Z)-2-((5-bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-dimethylami-
no-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
24 (71 mg, yield 77%) as a red solid.
[0272] MS m/z (ESI): 457.0[M+1]
[0273] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.663 (s, 1H,
pyrrole-NH), 11.011 (s, 1H, indole-NH), 8.118 (s, 1H, --ArH), 7.804
(s, 1H, --CH.dbd.C), 7.262.about.7.287 (m, 1H, --ArH),
6.826.about.6.847 (m, 1H, --ArH), 3.533.about.3.567 (t, 2H, N
seven-membered ring-CH.sub.2), 3.336.about.3.364 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.909.about.2.945 (t, 2H,
--CH.sub.2C.dbd.C), 2.465 (s, 3H, pyrrole-CH.sub.3),
2.401.about.2.434 (t, 2H, --CH.sub.2N), 2.204 (s, 6H,
2.times.-CH.sub.3N), 2.035.about.2.079 (m, 2H, seven-membered ring
intra-CH.sub.2).
Example 25
(Z)-5-(2-Dimethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-yliden-
emethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00129##
[0275] The title compound was prepared under the same conditions as
described in step 4 of Example 23 with
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c obtained from step 3 of Example 23
and 5-fluoro-1,3-dihydro-indol-2-one as starting materials to
obtain
(Z)-5-(2-dimethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-ylide-
nemethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
25 (205 mg, yield 68%) as a red solid.
[0276] MS m/z (ESI): 397.0[M+1]
[0277] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.714 (s, 1H,
pyrrole-NH), 10.904 (s, 1H, indole-NH), 7.760.about.7.783 (m, 1H,
--ArH), 7.744 (s, 1H, --CH.dbd.C), 6.915.about.6.943 (m, 1H,
--ArH), 6.836.about.6.868 (m, 1H, --ArH), 3.533.about.3.567 (t, 2H,
N seven-membered ring-CH.sub.2), 3.336.about.3.364 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.909.about.2.945 (t, 2H,
--CH.sub.2C.dbd.C), 2.457 (s, 3H, pyrrole-CH.sub.3),
2.401.about.2.434 (t, 2H, --CH.sub.2N), 2.204 (s, 6H,
2.times.-CH.sub.3N), 2.035.about.2.079 (m, 2H, seven-membered ring
intra-CH.sub.2).
Example 26
(Z)--N-{3-[5-(2-Dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro--
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-7--
yl}-formamide
##STR00130##
[0279] The title compound was prepared under the same conditions as
described in step 4 of Example 23 with
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c obtained from step 3 of Example 23
and N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-7-yl)-formamide 20c
obtained from step 3 of Example 20 as starting materials to obtain
(Z)--N-{3-[5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-
-pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-7-
-yl}-formamide 26 (71 mg, yield 79%) as a red solid.
[0280] MS m/z (ESI): 440.1[M+1]
[0281] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.653 (s, 1H,
pyrrole-NH), 10.437 (s, 1H, indole-NH), 9.814 (s, 1H, --NHCO),
8.339 (s, 1H, --CH.dbd.O), 7.768 (s, 1H, --CH.dbd.C), 7.621-7.649
(m, 1H, --ArH), 7.440.about.7.473 (m, 1H, --ArH), 3.533.about.3.567
(t, 2H, N seven-membered ring-CH.sub.2), 3.336.about.3.364 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 2.919.about.2.955 (t,
2H, --CH.sub.2C.dbd.C), 2.463 (s, 3H, pyrrole-CH.sub.3),
2.403.about.2.436 (t, 2H, --CH.sub.2N), 2.204 (s, 6H,
2.times.-CH.sub.3N), 2.035.about.2.079 (m, 2H, seven-membered ring
intra-CH.sub.2).
Example 27
(Z)--N-{3-[5-(2-Dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro--
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6--
yl}-2-hydroxy-acetamide
##STR00131##
##STR00132##
[0282] Step 1
Acetic acid
(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-ylcarbamoyl)-methyl
ester
[0283] 6-Amino-5-fluoro-1,3-dihydro-indol-2-one 3d (500 mg, 3.0
mmol) was dissolved in 10 ml of tetrahydrofuran under stirring, and
added with 0.4 ml of pyridine to the solution at room temperature.
After stirring to mix well, the mixture was cooled down to
-40.degree. C. in a dry ice-acetone bath. A solution of acetic acid
chlorocarbonylmethyl ester (420 mg, 3.0 mmol) in 10 ml of
tetrahydrofuran was added dropwise to the above reaction system.
Upon completion of the addition, the dry ice-acetone bath was
removed, and the reaction mixture was allowed to warm up to room
temperature and stirred overnight. After thin lay chromatography
showed the disappearance of starting materials, the reaction
mixture was filtered. The resulting solid was washed with water for
three times and dried to obtain the title compound acetic acid
(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-ylcarbamoyl)-methyl ester
27a (562 mg, yield 70.4%) as a gray solid.
[0284] MS: 265.3[M-1]
Step 2
N-(5-Fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-acetamide
[0285] Acetic acid
(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-ylcarbamoyl)-methyl ester
27a (58 mg, 0.22 mmol) was dissolved in 1 ml of methanol under
stirring, and added with 1 ml of water and sodium hydroxide (15 mg,
0.375 mmol) to the solution at room temperature. Upon completion of
the addition, the reaction mixture was stirred for another 1 hour.
After thin lay chromatography showed the disappearance of starting
materials, the reaction mixture was filtered. The resulting solid
was washed with water for three times and dried to obtain the title
compound
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-acetamide
27b (46 mg, yield 93.8%) as a gray solid.
[0286] MS: 223.7 [M-1]
Step 3
(Z)--N-{3-[5-(2-Dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro--
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6--
yl}-2-hydroxy-acetamide
[0287] The title compound was prepared under the same conditions as
described in step 4 of Example 23 with
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c obtained from step 3 of Example 23
and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-acetamide
27b as starting materials to obtain
(Z)--N-{3-[5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-
-pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-
-yl}-2-hydroxy-acetamide 27 (80 mg, yield 83.4%) as an orange
solid.
[0288] MS m/z (ESI): 470.1[M+1]
[0289] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.599 (s, 1H,
pyrrole-NH), 10.893 (s, 1H, indole-NH), 9.233 (s, 1H, --NHCO),
7.854.about.7.879 (m, 1H, --ArH), 7.725 (s, 1H, --CH.dbd.C),
7.663.about.7.688 (m, 1H, --ArH), 5.950 (s, 1H, --OH), 4.053 (s,
2H, --CH.sub.2O), 3.544 (t, 2H, N seven-membered ring-CH.sub.2),
3.315.about.3.340 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 2.924 (t, 2H, --CH.sub.2C.dbd.C), 2.464 (t, 2H,
--CH.sub.2N), 2.442 (s, 3H, pyrrole-CH.sub.3), 2.199 (s, 6H,
2.times.-CH.sub.3N), 2.044 (m, 2H, seven-membered ring
intra-CH.sub.2).
Example 28
(Z)-2-[4-(2,3-Difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-3--
methyl-5-(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]aze-
pin-4-one
##STR00133##
##STR00134##
[0290] Step 1
3-Methyl-5-[3-(2-pyrrolidin-1-yl-ethylamino)-propyl]-1H-pyrrole-2,4-dicarb-
oxylic acid 2-tert-butyl ester 4-ethyl ester
[0291]
5-(3-Methanesulfonyloxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxyli-
c acid 2-tert-butyl ester 4-ethyl ester 1g (8.462 g, 21.75 mmol)
was dissolved in 2-pyrrolidin-1-yl-ethylamine (6.3 ml, 49.79) under
stirring, the resulting solution was stirred at room temperature
overnight. After thin lay chromatography showed the disappearance
of starting materials, the reaction mixture was added with ethyl
acetate (200 ml) and a little methanol until a clear solution was
obtained. The mixture was washed with water (30 ml.times.3),
saturated brine (40 ml.times.2) and concentrated under reduced
pressure to obtain light brown oil. The oil was purified by silica
gel column chromatography to obtain the title compound
3-methyl-5-[3-(2-pyrrolidin-1-yl-ethylamino)-propyl]-1H-pyrrole-2,4-dicar-
boxylic acid 2-tert-butyl ester 4-ethyl ester 28a (4.488 g, yield
63.5%) as a yellow oil.
[0292] MS m/z (ESI): 406.5[M-1]
Step 2
3-Methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carboxylic acid tert-butyl ester
[0293]
3-Methyl-5-[3-(2-pyrrolidin-1-yl-ethylamino)-propyl]-1H-pyrrole-2,4-
-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester 28a (6.754 g,
16.6 mmol) was dissolved in 150 ml of toluene under stirring, and
added slowly with a solution of trimethyl aluminum in toluene (16.6
ml, 2 mol/L, 33.2 mmol) to the solution under an argon atmosphere.
The reaction mixture was stirred for 20 minutes at room temperature
until no white smoke was released, and refluxed for another 3.5
hours in an oil bath. After thin lay chromatography showed the
disappearance of starting materials, the oil bath was removed. The
reaction mixture was quenched with little ethanol (95%), added with
ethyl acetate (100 ml), filtered through a pad of Celite and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography to obtain the title compound
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carboxylic acid tert-butyl ester 28b (3.894 g,
yield 65%) as a yellow oil.
[0294] MS m/z (ESI): 362.2[M+1]
Step 3
3-Methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde
[0295]
3-Methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-py-
rrolo[3,2-c]azepine-2-carboxylic acid tert-butyl ester 28b (3.562
g, 9.87 mmol) was dissolved in 50 ml of dichloromethane under
stirring, and added with trifluoroacetic acid (19.7 ml, 260 mmol)
to the solution at room temperature. Upon the completion of the
addition, the mixture was heated to reflux for 30 minutes in an oil
bath. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was cooled down to
-5.degree. C. in an ice-salt bath, added with triethoxy methane
(2.96 ml, 14.8 mmol) in one portion, stirred at -5.degree. C. for 5
minutes, for another 1 hour at room temperature. The reaction
system was added with water (25 ml), adjusted to about pH 11 with
dilute sodium hydroxide solution (2 mol/L) and extracted with
dichloromethane (100 ml.times.3). The combined organic extracts
were concentrated under reduced pressure to obtain a henna oil. The
oil was purified by silica gel column chromatography to obtain the
title compound
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c (1.116 g, yield 49%) as a yellow
solid.
[0296] MS m/z (ESI): 290.2[M+1]
Step 4
(Z)-2-[4-(2,3-Difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-3--
methyl-5-(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]aze-
pin-4-one
[0297]
3-Methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-py-
rrolo[3,2-c]azepine-2-carbaldehyde 28c (40 mg, 0.134 mmol) and
4-(2,3-difluoro-phenyl)-1,3-dihydro-indol-2-one 6d obtained from
step 3 of Example 6 were dissolved in 0.3 ml of methanol under
stirring, and added with piperidine (0.03 ml, 0.3 mmol) to the
solution. Upon the completion of the addition, the mixture was
stirred at room temperature overnight. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was concentrated under reduced pressure to obtain
the title compound
(Z)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-3-
-methyl-5-(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]az-
epin-4-one 28 (40 mg, yield 57%) as a yellow solid.
[0298] MS m/z (ESI): 517.2[M+1]
[0299] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.532 (s, 1H,
pyrrole-NH), 11.132 (s, 1H, indole-NH), 7.598.about.7.620 (m, 1H,
--ArH), 7.421.about.7.434 (m, 1H, --ArH), 7.307.about.7.323 (m, 1H,
--ArH), 7.232.about.7.271 (m, 1H, --ArH), 7.001.about.7.021 (d, 1H,
--ArH), 6.865.about.6.884 (d, 1H, --ArH), 6.698 (s, 1H,
--CH.dbd.C), 3.499.about.3.533 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 3.273.about.3.302 (t, 2H, N seven-membered
ring-CH.sub.2), 2.854.about.2.891 (t, 2H, --CH.sub.2C.dbd.C),
2.536.about.2.570 (t, 2H, --CH.sub.2N), 2.498.about.2.515 (m, 4H,
five-membered ring-CH.sub.2N), 1.982.about.2.012 (m, 2H,
seven-membered ring-CH.sub.2), 1.777 (s, 3H, pyrrole-CH.sub.3),
1.657 (m, 4H, five-membered ring-CH.sub.2).
Example 29
(Z)--N-{5-Fluoro-3-[3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-
-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol--
7-yl}-formamide
##STR00135##
[0301] The title compound was prepared under the same conditions as
described in step 4 of Example 28 with
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c obtained from step 3 of Example 28
and N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-7-yl)-formamide 20c
obtained from step 3 of Example 20 as starting materials to obtain
(Z)--N-{5-fluoro-3-[3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-
-7-yl}-formamide 29 (59 mg, yield 95%) as a yellow solid.
[0302] MS m/z (ESI): 466.2[M+1]
[0303] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.679 (s, 1H,
pyrrole-NH), 10.868 (s, 1H, indole-NH), 8.324 (s, 1H, --HCO), 7.796
(s, 1H, --ArH), 7.747 (s, 1H, --CH.dbd.C), 7.601.about.7.629 (dd,
1H, --ArH), 7.424.about.7.454 (dd, 1H, --ArH), 3.543.about.3.577
(t, 2H, amide N seven-membered ring outer-CH.sub.2),
3.330.about.3.357 (t, 2H, N seven-membered ring-CH.sub.2),
2.907.about.2.944 (t, 2H, --CH.sub.2C.dbd.C), 2.576.about.2.610 (t,
2H, --CH.sub.2N), 2.498.about.2.515 (m, 4H, five-membered
ring-CH.sub.2N), 2.449 (s, 3H, pyrrole-CH.sub.3), 2.026.about.2.055
(m, 2H, seven-membered ring-CH.sub.2), 1.679 (m, 4H, five-membered
ring-CH.sub.2).
Example 30
(Z)--N-{3-[3-Methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydr-
o-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-5-yl}-ace-
tamide
##STR00136##
##STR00137##
[0304] Step 1
5-Nitro-indol-2-one
[0305] 1,3-Dihydro-indol-2-one 30a (20.0 g, 150 mmol) was dissolved
in sulfuric acid (100 ml, 98%) in an ice-water bath under stirring,
and added dropwise with nitric acid (10 ml, 65%-68%) while
maintaining the temperature below 0.degree. C. Upon completion of
the addition, the mixture was stirred for 1 hour at 0.degree. C.
After thin lay chromatography showed the disappearance of starting
materials, the reaction mixture was added with ice and filtered
after ice-out. The filter cake was washed with water (20
ml.times.3), and the resulting solid was recrystallized to obtain
the title compound 5-nitro-indol-2-one 30b (25.3 g, yield 92.4%) as
an orange solid.
[0306] MS m/z (ESI): 177.3[M-1]
Step 2
5-Amino-1,3-dihydro-indol-2-one
[0307] 5-Nitro-1,3-dihydro-indol-2-one 30b (3.56 g, 20 mmol) was
dissolved in 200 ml of acetic acid under stirring, and added with
palladium on activated carbon (1.0 g, 5%) to the solution at room
temperature. The reaction mixture was stirred under a hydrogen
atmosphere. After thin lay chromatography showed the disappearance
of starting materials, the reaction mixture was filtered, and
concentrated under reduced pressure to obtain the title compound
5-amino-1,3-dihydro-indol-2-one 30c (2.04 g, yield 68.9%) as a
white solid.
[0308] MS m/z (ESI): 149.4[M+1]
Step 3
N-(2-Oxo-2,3-dihydro-1H-indol-5-yl)-acetamide
[0309] 5-Amino-1,3-dihydro-indol-2-one 30c (3.5 g, 23.6 mmol) was
dissolved in 20 ml of tetrahydrofuran under stirring, and added
with triethylamine (3.6 ml, 26 mmol) to the solution at room
temperature. Upon completion of the addition, the mixture was
cooled down to -30.degree. C. in a dry ice-acetone bath, and added
slowly with acetyl chloride (1.8 ml, 24.8 mmol) while maintaining
the temperature below -20.degree. C. Upon completion of the
addition, the dry ice-acetone bath was removed, and the reaction
mixture was allowed to warm up to room temperature and stirred for
20 minutes. After thin lay chromatography showed the disappearance
of starting materials, the reaction mixture was added with ethyl
acetate (20 ml), gray solids were formed and filtered. The filter
cake was washed with water (70 ml.times.3) to obtain 2.5 g of
solids. The filtrate was extracted with ethyl acetate (200
ml.times.3). The combined organic extracts were concentrated under
reduced pressure, combined with the above solids to obtain the
title compound N-(2-oxo-2,3-dihydro-1H-indol-5-yl)-acetamide 30d
(4.0 g, yield 89%) as a gray solid.
[0310] MS m/z (ESI): 191.2[M+1]
Step 4
(Z)--N-{3-[3-Methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydr-
o-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-5-yl}-ace-
tamide
[0311] The title compound was prepared under the same conditions as
described in step 4 of Example 28 with
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c obtained from step 3 of Example 28
and N-(2-oxo-2,3-dihydro-1H-indol-5-yl)-acetamide 30d as starting
materials to obtain
(Z)--N-{3-[3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7-
,8-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indo-
l-5-yl}-acetamide 30 (50 mg, yield 80%) as a yellow solid.
[0312] MS m/z (ESI): 462.2[M+1]
[0313] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.679 (s, 1H,
pyrrole-NH), 10.868 (s, 1H, indole-NH), 9.806 (s, 1H, --NHCO),
7.796 (s, 1H, --ArH), 7.447 (s, 1H, --CH.dbd.C), 7.231.about.7.256
(dd, 1H, --ArH), 6.789.about.6.710 (s, 1H, --ArH),
3.513.about.3.547 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 3.303.about.3.330 (t, 2H, N seven-membered
ring-CH.sub.2), 2.866.about.2.903 (t, 2H, --CH.sub.2C.dbd.C),
2.540.about.2.574 (1, 2H, --CH.sub.2N), 2.461.about.2.513 (m, 4H,
five-membered ring-CH.sub.2N), 2.388 (s, 3H, pyrrole-CH.sub.3),
2.002.about.2.024 (m, 2H, seven-membered ring-CH.sub.2), 2.024 (s,
3H, --CH.sub.3), 1.648 (m, 4H, five-membered ring --CH.sub.2).
Example 31
(Z)--N-{5-Fluoro-3-[3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-
-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol--
6-yl}-2-hydroxy-acetamide
##STR00138##
[0315] The title compound was prepared under the same conditions as
described in step 4 of Example 28 with
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c obtained from step 3 of Example 28
and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-acetamide
27b obtained from step 2 of Example 27 as starting materials to
obtain
(Z)--N-{5-fluoro-3-[3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-
-6-yl}-2-hydroxy-acetamide 31 (50 mg, yield 76%) as a yellow
solid.
[0316] MS m/z (ESI): 496.2[M+1]
[0317] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.658 (s, 1H,
pyrrole-NH), 10.857 (s, 1H, indole-NH), 9.426 (s, 1H, --NHCO),
7.826.about.7.854 (d, 1H, --ArH), 7.694.about.7.710 (d, 1H, --ArH),
7.640 (s, 1H, --CH.dbd.C), 5.717 (s, 1H, --HO), 4.035 (d, 2H,
--CH.sub.2O), 3.536.about.3.570 (t, 2H, N seven-membered
ring-CH.sub.2), 3.339 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 2.889.about.2.926 (t, 2H, --CH.sub.2C.dbd.C),
2.562.about.2.596 (t, 2H, --CH.sub.2N), 2.483.about.2.513 (m, 4H,
five-membered ring --CH.sub.2N), 2.427 (s, 3H, pyrrole-CH.sub.3),
2.034 (m, 2H, seven-membered ring-CH.sub.2), 1.673 (m, 4H,
five-membered ring-CH.sub.2).
Example 32
(Z)-2-((5-Fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-pi-
peridin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00139##
##STR00140## ##STR00141##
[0318] Step 1
3-Methyl-5-[3-(2-piperidin-1-yl-ethylamino)-propyl]-1H-pyrrole-2,4-dicarbo-
xylic acid 2-tert-butyl ester 4-ethyl ester
[0319]
5-(3-Methanesulfonyloxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxyli-
c acid 2-tert-butyl ester 4-ethyl ester 1g (10.64 g, 27.3 mmol) was
dissolved in 2-piperidin-1-yl-ethylamine (7 ml, 49.2 mmol), the
reaction solution was stirred at room temperature overnight. After
thin lay chromatography showed the disappearance of starting
materials, the reaction mixture was added with ethyl acetate (200
ml) and a little methanol until a clear solution was obtained. The
mixture was washed with water (30 ml.times.3), the organic phase
was washed with saturated brine (40 ml.times.2), and concentrated
under reduced pressure. The residue was purified by silica gel
column chromatography to obtain the title compound
3-methyl-5-[3-(2-piperidin-1-yl-ethylamino)-propyl]-1H-pyrrole-2,4-dicarb-
oxylic acid 2-tert-butyl ester 4-ethyl ester 32a (5.35 g, yield
46.5%) as a yellow oil.
[0320] MS m/z (ESI): 422.3[M+1]
Step 2
3-Methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carboxylic acid tert-butyl ester
[0321]
3-Methyl-5-[3-(2-piperidin-1-yl-ethylamino)-propyl]-1H-pyrrole-2,4--
dicarboxylic acid 2-tert-butyl ester 4-ethyl ester 32a (225 mg,
0.534 mmol) was dissolved in 5 ml of dry toluene under stirring,
the reaction system was cooled in an ice-water bath, and added with
a solution of trimethyl aluminum in toluene (0.534 ml, 2 mol/L,
1.07 mmol) under an argon atmosphere. Upon completion of the
addition, the reaction system was stirred for 20 minutes at room
temperature until no white smoke was released, refluxed for another
3 hours in an oil bath. After thin lay chromatography showed the
disappearance of starting materials, the oil bath was removed, the
reaction mixture was added with saturated brine (10 ml) and ethyl
acetate (20 ml), stirred for 15 minutes at room temperature and
filtered. The filter cake was washed with ethyl acetate (10
ml.times.3). The filtrate was extracted with ethyl acetate (10
ml.times.2). The combined organic extracts were washed with
saturated brine (10 ml.times.2), dried over anhydrous magnesium
sulfate, filtered to remove the drying agent and concentrated under
reduced pressure to obtain the title compound
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carboxylic acid tert-butyl ester 32b (105 mg) as a
colorless oil to be used directly in the next step.
[0322] MS m/z (ESI): 376.2[M+1]
Step 3
3-Methyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]az-
epin-4-one
[0323]
3-Methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyr-
rolo[3,2-c]azepine-2-carboxylic acid tert-butyl ester 32b (953 mg,
2.54 mmol) was dissolved in 3 ml of ethanol under stirring, and
added dropwise with hydrochloric acid (3.2 ml, 12 mol/L) in an
ice-water bath under an argon atmosphere. Upon completion of the
addition, the ice-water bath was removed, and the reaction mixture
was stirred at 60.degree. C. in an oil bath for 1 hour. After thin
lay chromatography showed the disappearance of starting materials,
the reaction mixture was adjusted to about pH 7 with aqueous sodium
hydroxide solution (10 mol/L) and concentrated under reduced
pressure to evaporate ethanol. The residue was adjusted to pH 10
with aqueous sodium hydroxide solution (10 mol/L), extracted with
dichloromethane (20 ml.times.3). The combined organic extracts were
washed with saturated brine (20 ml), dried over anhydrous magnesium
sulfate, filtered to remove the drying agent and concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to obtain the title compound
3-methyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]a-
zepin-4-one 32c (395 m, yield 57%) as a white solid.
[0324] MS m/z (ESI): 276.1[M+1]
Step 4
3-Methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde
[0325] Dichloromethane (36 ml, 559 mmol) and N,N-dimethylformamide
(1.637 ml, 20.9 mmol) were stirred for 5 minutes at -15.degree. C.
in an ice-salt bath under an argon atmosphere. The solution was
added dropwise with phosphorus oxychloride (1.07 ml, 11.5 mmol) and
stirred for 15 minutes while maintaining the temperature at
-10.degree. C.
3-methyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]a-
zepin-4-one 32c (1.26 g, 4.58 mmol) was dissolved in 10 ml of
dichloromethane, the resulting solution was added dropwise to the
above solution. Upon completion of the addition, the ice-salt bath
was removed, and the reaction mixture was stirred for 3 hours at
room temperature. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
quenched with iced water, adjusted to about pH 10 with sodium
hydroxide solution (10 mol/L) and stirred for 30 minutes, extracted
with dichloromethane (30 ml.times.3). The combined organic extracts
were washed with saturated brine (30 ml), dried over anhydrous
magnesium sulfate, filtered to remove the drying agent and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography to obtain the title compound
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d (993 mg, yield 71.4%) as a light
yellow solid.
[0326] MS m/z (ESI): 304.1[M+1]
Step 5
(Z)-2-((5-Fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-pi-
peridin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
[0327]
3-Methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyr-
rolo[3,2-c]azepine-2-carbaldehyde 32d (50 mg, 0.165 mmol) and
5-fluoro-1,3-dihydro-indol-2-one (22.4 mg, 0.15 mmol) were
dissolved in 0.3 ml of ethanol under stirring, and added with
piperidine (0.05 ml, 0.5 mmol) to the solution at room temperature.
Upon completion of the addition, the mixture was stirred at
40.about.50.degree. C. in an oil bath for 5 hours. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was filtered to obtain yellow solid. The solid was
dissolved in ethanol (2 ml), heated to reflux for 30 minutes,
cooled down to room temperature and filtered to obtain the title
compound
(Z)-2-((5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-meth-
yl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-
-one 32 (38 g, yield 58%) as a yellow solid.
[0328] MS m/z (ESI): 437.1[M+1]
[0329] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.720 (s, 1H,
pyrrole-NH), 10.900 (s, 1H, indole-NH), 7.749.about.7.779 (dd, 1H,
--ArH), 7.740 (s, 1H, --CH.dbd.C), 6.912.about.6.963 (m, 1H,
--ArH), 6834.about.6.867 (d, 1H, --ArH), 3.539.about.3.572 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.326.about.3.3540, 2H, amide
N seven-membered ring outer-CHO, 2.922.about.2.958 (t, 2H,
seven-membered ring intra-CH.sub.2C.dbd.C), 2.452 (s, 3H,
pyrrole-CH.sub.3), 2.386.about.2.431 (m, 6H, 3.times.-CH.sub.2N),
2.027.about.2.091 (t, 2H, seven-membered ring intra-CH.sub.2),
1.474.about.1.499 (m, 4H, six-membered ring 2.times.-CH.sub.2),
1.379.about.1.391 (m, 2H, six-membered ring-CH.sub.2).
Example 33
(Z)-2-[4-(2,3-Difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-3--
methyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azep-
in-4-one
##STR00142##
[0331] The title compound was prepared under the same conditions as
described in step of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and 4-(2,3-difluoro-phenyl)-1,3-dihydro-indol-2-one 6d obtained
from step 3 of Example 6 as starting materials to obtain
(Z)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-3-
-methyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]aze-
pin-4-one 33 (51 mg, yield 80.9%) as a yellow solid.
[0332] MS m/z (ESI): 531.2[M+1]
[0333] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.540 (s, 1H,
pyrrole-NH), 11.130 (s, 1H, indole-NH), 7.618.about.7.640 (m, 1H,
--ArH), 7.429.about.7.461 (m, 1H, --ArH), 7.306.about.7.360 (m, 1H,
--ArH), 7.232-7.2710 (m, 1H, --ArH), 6.998.about.7.017 (d, 1H,
--ArH), 6.874.about.6.893 (d, 1H, --ArH), 6.709 (s, 1H, 3.504-3.536
(t, 2H, seven-membered ring intra-CH.sub.2N), 3.289.about.3.312 (t,
2H, amide N seven-membered ring outer-CH.sub.2), 2.887.about.2.923
(t, 2H, seven-membered ring intra-CH.sub.2C.dbd.C),
2.365.about.2.417 (m, 6H, 3.times.-CH.sub.2N), 2.009.about.2.038
(t, 2H, --CH.sub.2), 1.792 (s, 3H, pyrrole-CH.sub.3),
1.456.about.1.468 (m, 4H, six-membered ring-2.times.-CH.sub.2),
1.368.about.1.377 (m, 2H, six-membered ring-CH.sub.2).
Example 34
(Z)-2-[5-(4-Methoxy-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-3-met-
hyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin--
4-one
##STR00143##
[0335] The title compound was prepared under the same conditions as
described in step 5 of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and 5-(4-methoxy-phenyl)-1,3-dihydro-indol-2-one as starting
materials to obtain
(Z)-2-[5-(4-methoxy-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethy-
l]-3-methyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c-
]azepin-4-one 34 (68 mg, yield 68.7%) as a yellow solid.
[0336] MS m/z (ESI): 525.2[M+1]
[0337] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.653 (s, 1H,
pyrrole-NH), 11.003 (s, 1H, indole-NH), 7.930.about.7.850 (d, 1H,
--ArH), 7.668 (s, 1H, --CH.dbd.C), 7.583.about.7.605 (d, 2H,
--ArH), 7.251.about.7.274 (d, 1H, --ArH), 7.017.about.7.071 (m, 3H,
--ArH), 3.804 (s, 3H, --CH.sub.3O), 3.541.about.3.574 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.343 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.926.about.2.962 (t, 2H,
seven-membered ring intra-CH.sub.2C.dbd.C), 2.386.about.2.449 (m,
9H, pyrrole-CH.sub.3, 3.times.-CH.sub.2N), 2.062 (m, 2H,
seven-membered ring intra-CH.sub.2), 1.475.about.1.487 (m, 4H,
six-membered ring-2.times.-CH.sub.2), 1.379.about.1.391 (m, 2H,
six-membered ring-CH.sub.2).
Example 35
(Z)-2-((5-Chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-pi-
peridin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00144##
[0339] The title compound was prepared under the same conditions as
described in step 5 of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and 5-chloro-1,3-dihydro-indol-2-one as starting materials to
obtain
(Z)-2-((5-chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-p-
iperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
35 (44 mg, yield 64.8%) as a yellow solid.
[0340] MS m/z (ESI): 453.2[M+1]
[0341] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.674 (s, 1H,
pyrrole-NH), 11.003 (s, 1H, indole-NH), 7.989 (s, 1H, --ArH), 7.799
(s, 1H, --CH.dbd.C), 7.135.about.7.155 (d, 1H, --ArH),
6869.about.6.890 (d, 1H, --ArH), 3.545.about.3.576 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.313.about.3.342 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 2.925.about.2.962 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.399.about.2.459
(m, 9H, pyrrole-CH.sub.3, 3.times.-CH.sub.2N), 2.047.about.2.074
(1, 2H, seven-membered ring intra-CH.sub.2), 1.490 (m, 4H,
six-membered ring-2.times.-CH.sub.2), 1.385 (m, 2H, six-membered
ring-CH.sub.2).
Example 36
(Z)-2-((5-Bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-pip-
eridin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00145##
[0343] The title compound was prepared under the same conditions as
described in step of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and 5-bromo-1,3-dihydro-indol-2-one as starting materials to obtain
(Z)-2-((5-bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-pi-
peridin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
36 (43 mg, yield 57.6%) as a yellow solid.
[0344] MS m/z (ESI): 497.2[M+1]
[0345] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.670 (s, 1H,
pyrrole-NH), 11.010 (s, 1H, indole-NH), 8.114 (s, 1H, --ArH), 7.802
(s, 1H, --CH.dbd.C), 7.262.about.7.283 (d, 1H, --ArH),
6.826.about.6.846 (d, 1H, --ArH), 3.542.about.3.574 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.315.about.3.339 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 2.924.about.2.960 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.389.about.2.460
(m, 9H, pyrrole-CH.sub.3, 3.times.-CH.sub.2N), 2.045.about.2.074
(t, 2H, seven-membered ring intra-CH.sub.2), 1.476.about.1.489 (m,
4H, six-membered ring-2.times.-CH.sub.2), 1.381 (m, 2H,
six-membered ring-CH.sub.2).
Example 37
(Z)--N-{5-Fluoro-3-[3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8--
hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-6-
-yl}-2-methoxy-acetamide
##STR00146##
[0347] The title compound was prepared under the same conditions as
described in step of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-methoxy-acetamide 7a
obtained from step 1 of Example 7 as starting materials to obtain
(Z)--N-{5-fluoro-3-[3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-
-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol--
6-yl}-2-methoxy-acetamide 37 (59 mg, yield 75%) as a yellow
solid.
[0348] MS m/z (ESI): 524.2[M+1]
[0349] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.613 (s, 1H,
pyrrole-NH), 10.890 (s, 1H, indole-NH), 9.316 (s, 1H, --NHCO),
7.835.about.7.863 (d, 1H, --ArH), 7.668 (s, 1H, --CH.dbd.C),
7.542.about.7.558 (d, 1H, --ArH), 4.064 (s, 2H, --CH.sub.2O),
3.540.about.3.572 (t, 2H, seven-membered ring intra-CH.sub.2N),
3.406 (s, 3H, CH.sub.3O), 3.315.about.3.340 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.918.about.2.954 (t, 2H,
seven-membered ring intra-CH.sub.2C.dbd.C), 2.390.about.2.467 (m,
9H, pyrrole-CH.sub.3, 3.times.-CH.sub.2N), 2.043.about.2.072 (t,
2H, seven-membered ring intra-CH.sub.2), 1.478.about.1.490 (m, 4H,
six-membered ring-2.times.-CH.sub.2), 1.382.about.1.393 (m, 2H,
six-membered ring-CH.sub.2).
Example 38
(S,Z)--N-{5-Fluoro-3-[3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-
-6-yl}-2-hydroxy-propionamide
##STR00147##
[0351] The title compound was prepared under the same conditions as
described in step 5 of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-hydroxy-propionamide
8b obtained from step 2 of Example 8 as starting materials to
obtain
(S,Z)--N-{5-fluoro-3-[3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7-
,8-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indo-
l-6-yl}-2-hydroxy-propionamide 38 (49 mg, yield 62.5%) as a yellow
solid.
[0352] MS m/z (ESI): 524.2[M+1]
[0353] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.603 (s, 1H,
pyrrole-NH), 10.900 (s, 1H, indole-NH), 9.246 (s, 1H, --NH),
7.853.about.7.881 (d, 1H, --ArH), 7.726.about.7.743 (d, 1H, --ArH),
7.659 (s, 1H, --CH.dbd.C), 6.058.about.6.070 (d, 1H, --OH),
4.207.about.4.237 (m, 1H, --CHO), 3.541.about.3.573 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.326.about.3.354 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 2.917.about.2.954 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.392.about.2.440
(m, 9H, pyrrole-CH.sub.3, 3.times.-CH.sub.2N), 2.025.about.2.089
(t, 2H, seven-membered ring intra-CH.sub.2), 1.478.about.1.490 (m,
4H, six-membered ring-2.times.-CH.sub.2), 1.383 (m, 2H,
six-membered ring-CH.sub.2), 1.330.about.1.347 (d, 3H,
CH.sub.3O).
Example 39
(Z)--N-{3-[5-(2-Dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro--
pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6--
yl}-2-methoxy-acetamide
##STR00148##
[0355] The title compound was prepared under the same conditions as
described in step 4 of Example 23 with
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c obtained from step 3 of Example 23
and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-methoxy-acetamide 7a
obtained from step 1 of Example 7 as starting materials to obtain
(Z)--N-{3-[5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-
-pyrrolo[3,2-c]azepin-2-ylmethylene]-5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-
-yl}-2-methoxy-acetamide 39 (75 mg, yield 76.5%) as a brown
solid.
[0356] MS m/z (ESI): 484.1[M+1]
[0357] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.607 (s, 1H,
pyrrole-NH), 10.894 (s, 1H, --NH), 9.320 (s, 1H, --NHCO),
7.840.about.7.868 (d, 1H, --ArH), 7.673 (s, 1H, --CH.dbd.C),
7.540.about.7.557 (d, 1H, --ArH), 4.064 (s, 2H, --CH.sub.2O),
3.531.about.3.564 (t, 2H, N seven-membered ring-CH.sub.2), 3.406
(s, 3H, --CH.sub.3O), 3.333.about.3.359 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.904.about.2.941 (t, 2H,
--CH.sub.2C.dbd.C), 2.445 (s, 3H, pyrrole-CH.sub.3),
2.404.about.2.420 (t, 2H, --CH.sub.2N), 2.206 (s, 6H,
2.times.-CH.sub.3N), 2.029.about.2.057 (m, 2H, seven-membered ring
intra --CH.sub.2).
Example 40
(Z)-2-[4-(2,6-Difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-5--
(2-dimethylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepi-
n-4-one
##STR00149##
[0359] The title compound was prepared under the same conditions as
described in step 4 of Example 23 with
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c obtained from step 3 of Example 23
and 4-(2,6-difluoro-phenyl)-1,3-dihydro-indol-2-one as starting
materials to obtain
(Z)-2-[4-(2,6-difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylideneme-
thyl]-5-(2-dimeth
ylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
40 (36 mg, yield 36.4%) as an orange solid.
[0360] MS m/z (ESI): 491.1[M+1]
[0361] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.552 (s, 1H,
pyrrole-NH), 11.075 (s, 1H, indole-NH), 7.661 (m, 1H, --ArH),
7.338.about.7.378 (m, 2H, --ArH), 7.238.about.7.277 (m, 1H, --ArH),
7.008.about.7.027 (m, 1H, --ArH), 6.895.about.6.914 (d, 1H, --ArH),
6.652 (s, 1H, --CH.dbd.C), 3.488.about.3.522 (t, 2H, N
seven-membered ring-CH.sub.2), 3.280.about.3.316 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.868.about.2.904 (t, 2H,
--CH.sub.2C.dbd.C), 2.355.about.2.388 (t, 2H, --CH.sub.2N), 2.171
(s, 6H, 2.times.-CH.sub.3N), 1.992.about.2.021 (m, 2H,
seven-membered ring intra-CH.sub.2), 1.764 (s, 3H,
pyrrole-CH.sub.3).
Example 41
(Z)-5-(2-Dimethylamino-ethyl)-2-[4-(3-fluoro-phenyl)-2-oxo-1,2-dihydro-ind-
ol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4--
one
##STR00150##
[0363] The title compound was prepared under the same conditions as
described in step 4 of Example 23 with
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c obtained from step 3 of Example 23
and 4-(3-fluoro-phenyl)-1,3-dihydro-indol-2-one as starting
materials to obtain
(Z)-5-(2-dimethylamino-ethyl)-2-[4-(3-fluoro-phenyl)-2-oxo-1,2-dih-
ydro-indol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]a-
zepin-4-one 41 (37 mg, yield 38.9%) as a yellow solid.
[0364] MS m/z (ESI): 473.2[M+1]
[0365] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.533 (s, 1H,
pyrrole-NH), 11.075 (s, 1H, indole-NH), 7.595.about.7.610 (m, 1H,
--ArH), 7.286.about.7.343 (m, 3H, --ArH), 7.191.about.7.229 (m, 1H,
--ArH), 6.942.about.6.962 (d, 1H, --ArH), 6.830.about.6.844 (d, 1H,
--ArH), 6.811 (s, 1H, --CH.dbd.C), 3.488.about.3.521 (t, 2H, N
seven-membered ring-CH.sub.2), 3.277.about.3.315 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.859.about.2.896 (t, 2H,
--CH.sub.2C.dbd.C), 2.361.about.2.394 (t, 2H, --CH.sub.2N), 2.176
(s, 6H, 2.times.-CH.sub.3N), 1.989.about.2.018 (m, 2H,
seven-membered ring intra-CH.sub.2), 1.774 (s, 3H,
pyrrole-CH.sub.3).
Example 42
(Z)-2-[4-(2,3-Difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-5--
(2-dimethylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepi-
n-4-one
##STR00151##
[0367] The title compound was prepared under the same conditions as
described in step 4 of Example 23 with
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c obtained from step 3 of Example 23
and 4-(2,3-difluoro-phenyl)-1,3-dihydro-indol-2-one 6d obtained
from step 3 of Example 6 were used as starting materials to obtain
(Z)-2-[4-(2,3-difluoro-phenyl)-2-oxo-1,2-dihydro-indol-3-ylidenemethyl]-5-
-(2-dimethylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azep-
in-4-one 42 (25 mg, yield 25.2%) as a yellow solid.
[0368] MS m/z (ESI): 491.2[M+1]
[0369] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.532 (s, 1H,
pyrrole-NH), 11.132 (s, 1H, indole-NH), 7.618.about.7.640 (m, 1H,
--ArH), 7.429.about.7.442 (m, 1H, --ArH), 7.306.about.7.340 (m, 1H,
--ArH), 7.232.about.7.271 (m, 1H, --ArH), 6.998.about.7.017 (d, 1H,
--ArH), 6.874.about.6.893 (d, 1H, --ArH), 6.712 (s, 1H,
--CH.dbd.C), 3.491.about.3.525 (t, 2H, N seven-membered
ring-CH.sub.2), 3.292.about.3.315 (t, 2H, amide N seven-membered
ring outer-CH.sub.2), 2.869.about.2.905 (t, 2H, --CH.sub.2C.dbd.C),
2.362.about.2.395 (t, 2H, --CH.sub.2N), 2.176 (s, 6H,
2.times.-CH.sub.3N), 1.993.about.2.018 (m, 2H, seven-membered ring
intra-CH.sub.2), 1.795 (s, 3H, pyrrole-CH.sub.3).
Example 43
(Z)-5-(2-Dimethylamino-ethyl)-2-[5-fluoro-6-(4-fluoro-benzylamino)-2-oxo-1-
,2-dihydro-indol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3-
,2-c]azepin-4-one
##STR00152##
[0371] The title compound was prepared under the same conditions as
described in step 4 of Example 23 with
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c obtained from step 3 of Example 23
and 5-fluoro-6-(4-fluoro-benzylamino)-1,3-dihydro-indol-2-one 3e
obtained from step 4 of Example 3 were used as starting materials
to obtain
(Z)-5-(2-dimethylamino-ethyl)-2-[5-fluoro-6-(4-fluoro-benzylamino)-2-oxo--
1,2-dihydro-indol-3-ylidenemethyl]-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[-
3,2-c]azepin-4-one 43 (50 mg, yield 43.7%) as a red solid.
[0372] MS m/z (ESI): 520.1[M+1]
[0373] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.421 (s, 1H,
pyrrole-NH), 10.523 (s, 1H, indole-NH), 7.574.about.7.603 (d, 1H,
--ArH), 7.367.about.7.402 (m, 2H, --ArH), 7.513 (s, 1H,
--CH.dbd.C), 7.142.about.7.186 (m, 2H, --ArH), 6.41 (t, 1H, --NH),
6.041.about.6.059 (d, 1H, --ArH), 4.348.about.4.362 (d, 2H,
aniline-CH.sub.2), 3.982.about.3.996 (d, 2H, --CH.sub.2O),
3.531.about.3.564 (t, 2H, N seven-membered ring-CH.sub.2),
3.316.about.3.344 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 2.856.about.2.893 (t, 2H, --CH.sub.2C.dbd.C),
2.398.about.2.415 (t, 2H, --CH.sub.2N), 2.390 (s, 3H,
pyrrole-CH.sub.3), 2.192 (s, 6H, 2.times.-CH.sub.3N),
2.002.about.2.030 (m, 2H, seven-membered ring intra-CH.sub.2).
Example 44
(Z)--N-{3-[5-(2-Dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro--
pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-5-yl}-2-hyd-
roxy-acetamide
##STR00153##
##STR00154##
[0374] Step 1
Acetic acid (2-oxo-2,3-dihydro-1H-indol-5-ylcarbamoyl)-methyl
ester
[0375] 5-Amino-1,3-dihydro-indol-2-one 30c (500 mg, 3.38 mmol) was
dissolved in 10 ml of dichloromethane under stirring at room
temperature, and pyridine (470 .mu.l, 5 mmol) was added to the
solution at -40.degree. C. in a dry ice-acetone bath. Acetic acid
chlorocarbonylmethyl ester (473 mg, 3.48 mmol) was dissolved in 10
ml of dichloromethane, the resulting solution was added dropwise to
the above reaction solution. Upon completion of the addition, the
dry ice-acetone was removed, and the reaction mixture was allowed
to warm up to room temperature and stirred overnight. After thin
lay chromatography showed the disappearance of starting materials,
the reaction mixture was filtered. The filter cake was washed with
water (10 ml.times.3) and recrystallized to obtain the title
compound acetic acid
(2-oxo-2,3-dihydro-1H-indol-5-ylcarbamoyl)-methyl ester 44a (510
mg, yield 60.7%) as a yellow solid.
[0376] MS m/z (ESI): 247.7[M-1]
Step 2
2-Hydroxy-N-(2-oxo-2,3-dihydro-1H-indol-5-yl)-acetamide
[0377] Acetic acid
(2-oxo-2,3-dihydro-1H-indol-5-ylcarbamoyl)-methyl ester 44a (2.43
g, 10 mmol) was dissolved in 60 ml of methanol under stirring, and
sodium hydroxide solution (20 ml, 2 mol/L) was added to the
solution and stirred at room temperature overnight. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was neutralized with hydrochloric acid solution (6
mol/L) in ice-water bath, concentrated under reduced pressure. The
resulting solid was purified by silica gel column chromatography to
obtain the title compound
2-hydroxy-N-(2-oxo-2,3-dihydro-1H-indol-5-yl)-acetamide 44b (402
mg, yield 19.5%) as a yellow solid.
[0378] MS m/z (ESI): 205.3 [M-1]
Step 3
(Z)-2-Hydroxy-N-{3-[3-methyl-5-(2-methylamino-ethyl)-4-oxo-1,4,5,6,7,8-hex-
ahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-5-yl-
}-acetamide
[0379] The title compound was prepared under the same conditions as
described in step 4 of Example 23 with
5-(2-dimethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,-
2-c]azepine-2-carbaldehyde 23c obtained from step 3 of Example 23
and 2-hydroxy-N-(2-oxo-2,3-dihydro-1H-indol-5-yl)-acetamide 44b as
starting materials to obtain
(Z)-2-hydroxy-N-{3-[3-methyl-5-(2-methylamino-ethyl)-4-oxo-1,4,5,6,7,8-he-
xahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-5-y-
l}-acetamide 44 (52 mg, yield 50.5%) as an orange solid.
[0380] MS m/z (ESI): 452.1[M+1]
[0381] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.658 (s, 1H,
pyrrole-NH), 10.857 (s, 1H, --NH), 9.426 (s, 1H, NHCO),
7.939.about.7.942 (d, 1H, --ArH), 7.513 (s, 1H, --CH.dbd.C),
7.485.about.7.489 (d, 1H, --ArH), 6.820.about.6.841 (d, 1H, --ArH),
5.717 (s, 1H, --HO), 3.982.about.3.996 (d, 2H, --CH.sub.2O),
3.531.about.3.564 (t, 2H, N seven-membered ring-CH.sub.2),
3.337.about.3.365 (t, 2H, amide N seven-membered ring
outer-CH.sub.2), 2.908.about.2.944 (t, 2H, --CH.sub.2C.dbd.C),
2.433 (s, 3H, pyrrole-CH.sub.3), 2.394.about.2.411 (t, 2H,
--CH.sub.2N), 2.199 (s, 6H, 2.times.-CH.sub.3N), 2.031.about.2.059
(m, 2H, seven-membered ring intra-CH.sub.2).
Example 45
(Z)-2-((5-Fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-py-
rrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00155##
[0383] The title compound was prepared under the same conditions as
described in step 4 of Example 28 with
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c obtained from step 3 of Example 28
and 5-fluoro-1,3-dihydro-indol-2-one as starting materials to
obtain
(Z)-2-((5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-p-
yrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
45 (61 mg, yield 80.8%) as an orange solid.
[0384] MS m/z (ESI): 423.1[M+1]
[0385] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.715 (s, 1H,
pyrrole-NH), 10.902 (s, 1H, indole-NH), 7.752.about.7.782 (m, 1H,
--ArH), 7.743 (s, 1H, --CH.dbd.C), 6.937.about.6.965 (m, 1H,
--ArH), 6.835.about.6.867 (d, 1H, --ArH), 3.548.about.3.582 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.337.about.3.365 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 3.314 (m, 4H,
five-membered ring-2.times.-CH.sub.2N), 2.990.about.3.027 (t, 2H,
seven-membered ring intra-CH.sub.2C.dbd.C), 2.573.about.2.607 (t,
2H, --CH.sub.2N), 2.473 (s, 3H, pyrrole-CH.sub.3),
2.101.about.2.129 (m, 2H, seven-membered ring intra-CH.sub.2),
1.751 (m, 4H, five-membered ring-CH.sub.2).
Example 46
(Z)-2-((5-Bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-pyr-
rolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00156##
[0387] The title compound was prepared under the same conditions as
described in step 4 of Example 28 with
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c obtained from step 3 of Example 28
and 5-bromo-1,3-dihydro-indol-2-one as starting materials to obtain
(Z)-2-((5-bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-py-
rrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
46 (61 mg, yield 70.4%) as a yellow solid.
[0388] MS m/z (ESI): 485.1[M+1]
[0389] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.664 (s, 1H,
pyrrole-NH), 11.007 (s, 1H, indole-NH), 8.113.about.8.114 (d, 1H,
--ArH), 7.803 (s, 1H, --CH.dbd.C), 7.261.about.7.286 (m, 1H,
--ArH), 6.825.about.6.846 (d, 1H, --ArH), 3.548.about.3.582 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.337.about.3.364 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 3.312 (m, 4H,
five-membered ring-2.times.-CH.sub.2N), 2.905.about.2.942 (t, 2H,
seven-membered ring intra-CH.sub.2C.dbd.C), 2.573.about.2.606 (t,
2H, --CH.sub.2N), 2.462 (s, 3H, pyrrole-CH.sub.3),
2.028.about.2.057 (m, 2H, seven-membered ring intra-CH.sub.2),
1.751 (m, 4H, five-membered ring-CH.sub.2).
Example 47
(Z)-2-((5-Chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-py-
rrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00157##
[0391] The title compound was prepared under the same conditions as
described in step 4 of Example 28 with
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c obtained from step 3 of Example 28
and 5-chloro-1,3-dihydro-indol-2-one as starting materials to
obtain
(Z)-2-((5-chloro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-p-
yrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
47 (61 mg, yield 77.7%) as a yellow solid.
[0392] MS m/z (ESI): 439.2[M+1]
[0393] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.714 (s, 1H,
pyrrole-NH), 11.046 (s, 1H, indole-NH), 8.038 (s, 1H, --CH.dbd.C),
7.838.about.7.845 (d, 1H, --ArH), 7.179.about.7.205 (dd, 1H,
--ArH), 6.915.about.6.935 (d, 1H, --ArH), 3.612.about.3.629 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.384.about.3.412 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 3.337.about.3.384 (m,
4H, five-membered ring-2.times.-CH.sub.2N), 2.990.about.3.027 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.650.about.2.684
(t, 2H, --CH.sub.2N), 2.473 (s, 3H, pyrrole-CH.sub.3),
2.101.about.2.129 (m, 2H, seven-membered ring intra-CH.sub.2),
1.751 (m, 4H, five-membered ring-CH.sub.2).
Example 48
(Z)-3-Methyl-2-(2-oxo-5-phenyl-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-pyr-
rolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00158##
[0395] The title compound was prepared under the same conditions as
described in step 4 of Example 28 with
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c obtained from step 3 of Example 28
and 5-phenyl-1,3-dihydro-indol-2-one as starting materials to
obtain
(Z)-3-methyl-2-(2-oxo-5-phenyl-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-py-
rrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
48 (41 mg, yield 62.3%) as a yellow solid.
[0396] MS m/z (ESI): 481.2 [M+1]
[0397] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.706 (s, 1H,
pyrrole-NH), 10.070 (s, 1H, indole-NH), 8.174 (s, 1H, --CH.dbd.C),
7.843 (s, 1H, --ArH), 7.722.about.7.741 (d, 2H, --ArH),
7.443.about.7.480 (m, 3H, --ArH), 7.314.about.7.351 (t, 1H, --ArH),
6.961.about.6.981 (d, 1H, --ArH), 3.554.about.3.588 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.362.about.3.376 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 3.289.about.3.347 (m,
4H, five-membered ring-2.times.-CH.sub.2N), 2.990.about.3.027 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.650.about.2.684
(t, 2H, --CH.sub.2N), 2.473 (s, 3H, pyrrole-CH.sub.3),
2.101.about.2.129 (m, 2H, seven-membered ring intra-CH.sub.2),
1.751 (m, 4H, five-membered ring-2.times.-CH.sub.2).
Example 49
(Z)-2-(4-Bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-pyrr-
olidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00159##
[0399] The title compound was prepared under the same conditions as
described in step 4 of Example 28 with
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c obtained from step 3 of Example 28
and 4-bromo-1,3-dihydro-indol-2-one as starting materials to obtain
(Z)-2-(4-bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-pyr-
rolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
49 (40 mg, yield 60.5%) as a yellow solid.
[0400] MS m/z (ESI): 483.2[M-1]
[0401] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.697 (s, 1H,
pyrrole-NH), 11.244 (s, 1H, indole-NH), 8.650 (s, 1H, --CH--C),
7.281.about.7.301 (d, 1H, --ArH), 7.118.about.7.158 (m, 1H, --ArH),
7.000.about.7.190 (d, 1H, --ArH), 3.618.about.3.652 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.410.about.3.438 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 3.356.about.3.378 (m,
4H, five-membered ring-2.times.-CH.sub.2N), 2.990.about.3.027 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.650.about.2.684
(t, 2H, --CH.sub.2N), 2.473 (s, 3H, pyrrole-CH.sub.3),
2.101.about.2.129 (m, 2H, seven-membered ring intra-CH.sub.2),
1.751 (m, 4H, five-membered ring-CH.sub.2).
Example 50
(Z)-2-(7-Bromo-5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl--
5-(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-o-
ne
##STR00160##
[0403] The title compound was prepared under the same conditions as
described in step 4 of Example 28 with
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c obtained from step 3 of Example 28
and 7-bromo-5-fluoro-1,3-dihydro-indol-2-one 4b obtained from step
1 of Example 4 as starting materials to obtain
(Z)-2-(7-bromo-5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-
-5-(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4--
one 50 (51 mg, yield 73.2%) as an orange solid.
[0404] MS m/z (ESI): 501.1[M+1]
[0405] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.660 (s, 1H,
pyrrole-NH), 11.177 (s, 1H, indole-NH), 7.857.about.7.875 (d, 1H,
--ArH), 7.798 (s, 1H, --CH.dbd.C), 7.251.about.7.269 (d, 1H,
--ArH), 3.556.about.3.590 (t, 2H, seven-membered ring
intra-CH.sub.2N), 3.317.about.3.359 (t, 2H, amide N seven-membered
ring outer-CH.sub.2), 3.294.about.3.359 (m, 4H, five-membered
ring-2.times.-CH.sub.2N), 2.935.about.2.971 (t, 2H, seven-membered
ring intra-CH.sub.2C.dbd.C), 2.587.about.2.620 (t, 2H,
--CH.sub.2N), 2.473 (s, 3H, pyrrole-CH.sub.3), 2.059 (m, 2H,
seven-membered ring intra --CH.sub.2), 1.751 (m, 4H, five-membered
ring-CH.sub.2).
Example 51
(Z)--N-{3-[5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-p-
yrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-5-yl}-acetam-
ide
##STR00161##
[0407] The title compound was prepared under the same conditions as
described in step 10 of Example 1 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-
-c]azepine-2-carbaldehyde 1j obtained from step 9 of Example 1 and
N-(2-oxo-2,3-dihydro-1H-indol-5-yl)-acetamide 30d obtained from
step 3 of Example 30 as starting materials to obtain
(Z)--N-{3-[5-(2-diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro--
pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-5-yl}-aceta-
mide 51 (12 mg, yield 18.9%) as an orange solid.
[0408] MS m/z (ESI): 464.2[M+1]
[0409] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.679 (s, 1H,
pyrrole-NH), 10.868 (s, 1H, indole-NH), 9.806 (s, 1H, amide-NH),
7.841 (s, 1H, --ArH), 7.472 (s, 1H, --CH.dbd.C), 7.251.about.7.256
(d, 1H, --ArH), 6.806.about.6.827 (s, 1H, --ArH), 3.499 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.322.about.3.347 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 2.902.about.2.939 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.530.about.2.562
(m, 6H, 3.times.-CH.sub.2N), 2.423 (s, 3H, pyrrole-CH.sub.3),
2.029.about.2.051 (m, 2H, seven-membered ring intra-CH.sub.2),
2.029 (s, 3H, --CH.sub.3CO), 0.958.about.0.993 (t, 6H,
2.times.-CH.sub.3).
Example 52
(Z)--N-{5-Fluoro-3-[3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-
-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol--
6-yl}-2-methoxy-acetamide
##STR00162##
[0411] The title compound was prepared under the same conditions as
described in step 4 of Example 28 with
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[-
3,2-c]azepine-2-carbaldehyde 28c obtained from step 3 of Example 28
and
N-(5-fluoro-2-oxo-2,3-dihydro-1H-indol-6-yl)-2-methoxy-acetamide 7a
obtained from step 1 of Example 7 as starting materials to obtain
(Z)--N-{5-fluoro-3-[3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepin-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indol-
-6-yl}-2-methoxy-acetamide 52 (45 mg, yield 63.7%) as a yellow
solid.
[0412] MS m/z (ESI): 510.1[M+1]
[0413] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.652 (s, 1H,
pyrrole-NH), 10.936 (s, 1H, indole-NH), 9.362 (s, 1H, --NHCO),
7.882.about.7.910 (d, 1H, --ArH), 7.714 (s, 1H, --CH.dbd.C),
7.582.about.7.598 (d, 1H, --ArH), 4.110 (s, 2H, --CH.sub.2O),
3.597.about.3.631 (t, 2H, seven-membered ring intra-CH.sub.2N),
3.447 (s, 3H, --CH.sub.3O), 3.381.about.3.408 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 3.331.about.3.355 (m, 4H,
five-membered ring-2.times.-CH.sub.2N), 2.945.about.2.981 (t, 2H,
seven-membered ring intra-CH.sub.2C.dbd.C), 2.660 (m, 2H,
--CH.sub.2N), 2.489 (s, 3H, pyrrole-CH.sub.3), 2.069.about.2.099
(m, 2H, seven-membered ring intra-CH.sub.2), 1.731 (m, 4H,
five-membered ring-CH.sub.2).
Example 53
(R,Z)-2-(5-Fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-hydroxy-3--
morpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-
-4-one
##STR00163##
##STR00164##
[0414] Step 1
4-Oxiranylmethyl-morpholine
[0415] Morpholine 53a (8.712 ml, 0.1 mol) was dissolved in
tert-butanol (4.5 ml) under stirring at room temperature, and
(R)-(-)-1-chloro-2,3-epoxypropane (8.05 ml, 0.1 mol) was added
slowly to the solution at 0.degree. C. in an ice-water bath. Upon
completion of the addition, the ice-water bath was removed, and the
reaction mixture was allowed to warm up to room temperature and
stirred for 24 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was added
dropwise with a solution of potassium tert-butoxide in
tetrahydrofuran (60 ml, 1.67 mol/L, 100 mmol) while maintaining the
temperature below 10.degree. C. in an ice-water bath, gradually
turned from light yellow to white turbid solution and stirred for
30 minutes. After thin lay chromatography showed the disappearance
of starting materials, the reaction mixture was concentrated under
reduced pressure, added with water (50 ml), exacted with
dichloromethane (100 ml.times.2). The combined organic extracts
were washed with saturated brine (100 ml), dried over anhydrous
magnesium sulfate, filtered to remove the drying agent and
concentrated under reduced pressure to obtain the title compound
4-oxiranylmethyl-morpholine 53b (12.7 g, yield 88.8%) as a yellow
oil.
[0416] MS m/z (ESI): 144.4[M+1]
Step 2
1-Amino-3-morpholin-4-yl-propan-2-ol
[0417] 4-Oxiranylmethyl-morpholine 53b (6.3 g, 44 mmol) was added
slowly with ammonia (450 ml, 25%, 6.6 mol) while maintaining the
temperature below 0.degree. C. in an ice-water bath. Upon
completion of the addition, the reaction mixture was allowed to
warm up to room temperature and stirred for 18 hours. After thin
lay chromatography showed the disappearance of starting materials,
the reaction mixture was concentrated under reduced pressure to
obtain the title compound 1-amino-3-morpholin-4-yl-propan-2-ol 53c
(7 g, yield 99%) as a white solid.
[0418] MS m/z (ESI): 161.1[M+1]
Step 3
5-[3-(2-Hydroxy-3-morpholin-4-yl-propylamino)-propyl]-3-methyl-1H-pyrrole--
2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester
[0419]
5-(3-methanesulfonyloxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxyli-
c acid 2-tert-butyl ester 4-ethyl ester 1 g (1.13 g, 2.9 mmol) was
dissolved in 5.6 ml of dichloromethane under stirring, and
1-amino-3-morpholin-4-yl-propan-2-ol 53c (0.93 g, 5.8 mmol) was
added to the solution at room temperature. Upon completion of the
addition, the reaction mixture was stirred at room temperature
overnight and heated for 14 hours at 45.degree. C. in an oil bath.
After thin lay chromatography showed the disappearance of starting
materials, the reaction mixture was added with saturated brine (15
ml), exacted with dichloromethane (20 ml.times.3). The combined
organic extracts were concentrated under reduced pressure, purified
by silica gel column chromatography to obtain the title compound
5-[3-(2-hydroxy-3-morpholin-4-yl-propylamino)-propyl]-3-methyl-1H-pyrrole-
-2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester 53d (600
mg, yield 72.5%) as a colorless oil.
[0420] MS m/z (ESI): 454.2[M+1]
Step 4
5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrro-
lo[3,2-c]azepin-4-one
[0421]
5-[3-(2-hydroxy-3-morpholin-4-yl-propylamino)-propyl]-3-methyl-1H-p-
yrrole-2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester 53d
(580 mg, 1.28 mmol) was dissolved in 6 ml of toluene under
stirring, and trimethyl aluminum in toluene (1.9 ml, 2 mol/L, 3.84
mmol) was added dropwise in an ice-water bath under an argon
atmosphere. Upon completion of the addition, the ice-water bath was
removed, and the reaction mixture was heated to reflux for 24
hours. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was concentrated under
reduced pressure, added with hydrochloric acid solution (20 ml, 6
mol/L) and stirred for 20 minutes at room temperature. The mixture
was adjusted to about pH 12 with aqueous sodium hydroxide solution
(12 mol/L) in an ice-water bath and extracted with dichloromethane
(50 ml.times.2). The combined organic extracts were concentrated
under reduced pressure, purified by silica gel column
chromatography to obtain the title compound
5-(2-hydroxy-3-morpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrr-
olo[3,2-c]azepin-4-one 53e (300 mg, yield 57.6%) as a white
solid.
[0422] MS m/z (ESI): 308.2[M+1]
Step 5
5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-
-pyrrolo[3,2-c]azepine-2-carbaldehyde
[0423] (Chloromethylene)dimethylammoniumchloride (130 mg, 0.977
mmol) was dissolved in 3 ml of dichloromethane under stirring, the
resulting solution was cooled down to 0.degree. C. in ice-water
bath under an argon atmosphere.
5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrr-
olo[3,2-c]azepin-4-one 53e (300 mg, 0.977 mmol) was dissolved in 2
ml of dichloromethane, the resulting solution was added dropwise to
the above reaction system while maintaining the temperature below
0.degree. C. and stirred for 20 minutes. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was quenched with aqueous sodium hydroxide
solution (12 mol/L), added with saturated brine (10 ml), extracted
with the solvent mixture (V:V=10:1, 100 ml.times.3) of
dichloromethane and methanol. The combined organic extracts were
washed with saturated brine (100 ml), dried over anhydrous
magnesium sulfate, filtered to remove the drying agent and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography to obtain the title compound
5-(2-hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydr-
o-pyrrol o[3,2-c]azepine-2-carbaldehyde 53f (200 mg, yield 61%) as
a white solid.
[0424] MS m/z (ESI): 336.2[M+1]
Step 6
(R,Z)-2-(5-Fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-hydroxy-3--
morpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-
-4-one
[0425]
5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-he-
xahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 53f (50 mg, 0.149
mmol) was dissolved in 261 .mu.l of ethanol under stirring, and
added with 5-fluoro-1,3-dihydro-indol-2-one (20.28 mg, 0.134 mmol)
and piperidine (7.3 .mu.l, 0.074 mmol) to the solution at room
temperature. Upon completion of the addition, the reaction mixture
was stirred for 2 hours in dark at 80.degree. C. in an oil bath.
After thin lay chromatography showed the disappearance of starting
materials, the oil bath was removed, and the reaction mixture was
naturally cooled down to room temperature, filtered and dried to
obtain the title compound
(R,Z)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-hydroxy-3-
-morpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepi-
n-4-one 53 (40 g, yield 57%) as a yellow solid.
[0426] MS m/z (ESI): 469.2[M+1]
[0427] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.725 (s, 1H,
pyrrole-NH), 10.907 (s, 1H, indole-NH), 7.760.about.7.783 (m, 1H,
--ArH), 7.747 (s, 1H, --CH.dbd.C), 6.914.about.6.939 (m, 1H,
--ArH), 6.835.about.6.867 (m, 1H, --ArH), 4.719.about.4.731 (d, 1H,
--OH), 3.897 (m, 1H, --CHO), 3.749.about.3.792 (dd, 1H, amide N
seven-membered ring outer-CH.sub.2), 3.570.about.3.592 (t, 4H,
morpholine-2.times.-CH.sub.2O), 3.384.about.3.351 (t, 2H, N
seven-membered ring-CH.sub.2), 3.138.about.3.191 (dd, 1H, amide N
seven-membered ring outer-CH.sub.2), 2.917.about.2.953 (t, 2H,
--CH.sub.2C.dbd.C), 2.457 (s, 3H, pyrrole-CH.sub.3),
2.418.about.2.507 (m, 4H, --CH.sub.2N, morpholine-CH.sub.2N),
2.289.about.2.313 (t, 2H, morpholine-CH.sub.2N), 2.076 (m, 2H,
seven-membered ring-CH.sub.2).
Example 54
(Z)-3-Methyl-2-(2-oxo-4-pyridin-4-yl-1,2-dihydro-indol-3-ylidenemethyl)-5--
(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00165##
[0429] The title compound was prepared under the same conditions as
described in step of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and 4-pyridin-4-yl-1,3-dihydro-indol-2-one as starting materials to
obtain
(Z)-3-methyl-2-(2-oxo-4-pyridin-4-yl-1,2-dihydro-indol-3-ylidenemethyl)-5-
-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
54 (40 mg, yield 54%) as a yellow solid.
[0430] MS m/z (ESI): 496.2[M+1]
[0431] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.530 (s, 1H,
pyrrole-NH), 11.115 (s, 1H, indole-NH), 8.738.about.8.753 (d, 2H,
--CH.dbd.N), 7.498 (s, 2H, pyridine-CH.dbd.C), 7.219.about.7.258
(m, 1H, --ArH), 6.976.about.6.996 (d, 1H, --ArH), 6.808.about.6.830
(d, 1H, --ArH), 6.808 (s, 1H, --CH.dbd.C), 3.499.about.3.532 (t,
2H, seven-membered ring intra-CH.sub.2N), 3.270.about.3.298 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 2.879.about.2.916 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.361.about.2.413
(m, 6H, 3.times.-CH.sub.2N), 2.055.about.2.084 (t, 2H,
seven-membered ring intra-CH.sub.2), 1.732 (s, 3H,
pyrrole-CH.sub.3), 1.453.about.1.478 (m, 4H, six-membered
ring-2.times.-CH.sub.2), 1.365.about.1.377 (m, 2H, six-membered
ring-CH.sub.2).
Example 55
(Z)-2-[5-Fluoro-6-(4-fluoro-benzylamino)-2-oxo-1,2-dihydro-indol-3-ylidene-
methyl]methyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-
-c]azepin-4-one
##STR00166##
[0433] The title compound was prepared under the same conditions as
described in step of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and 5-fluoro-6-(4-fluoro-benzylamino)-1,3-dihydro-indol-2-one 3e
obtained from step 4 of Example 3 as starting materials to obtain
(Z)-2-[5-fluoro-6-(4-fluoro-benzylamino)-2-oxo-1,2-dihydro-indol-3-yliden-
emethyl]-3-methyl-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo-
[3,2-c]azepin-4-one 55 (77 mg, yield 90.8%) as a yellow solid.
[0434] MS m/z (ESI): 560.1[M+1]
[0435] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.425 (s, 1H,
pyrrole-NH), 10.520 (s, 1H, --NH), 7.572.about.7.602 (d, 1H,
--ArH), 7.349 (s, 1H, --CH.dbd.C), 7.367.about.7.402 (m, 2H,
--ArH), 7.141.about.7.186 (m, 2H, --ArH), 6.398.about.6.422 (m, 1H,
--NH), 6.040.about.6.059 (m, 1H, --ArH), 4.347.about.4.361 (d, 2H,
aniline-CH.sub.2), 3.524.about.3.557 (t, 2H, seven-membered ring
intra-CH.sub.2N), 3.314.about.3.337 (t, 2H, amide N seven-membered
ring outer-CH.sub.2), 2.869.about.2.906 (t, 2H, seven-membered ring
intra-CH.sub.2C.dbd.C), 2.390.about.2.467 (m, 9H, pyrrole-CH.sub.3,
3.times.-CH.sub.2N), 1.999.about.2.063 (t, 2H, seven-membered ring
intra-CH.sub.2), 1.476.about.1.489 (m, 4H, six-membered
ring-2.times.-CH.sub.2), 1.383.about.1.393 (m, 2H, six-membered
ring-CH.sub.2).
Example 56
(Z)-2-(7-Bromo-5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl--
5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-on-
e
##STR00167##
[0437] The title compound was prepared under the same conditions as
described in step of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and 7-bromo-5-fluoro-1,3-dihydro-indol-2-one 4b obtained from step
1 of Example 4 as starting materials to obtain
(Z)-2-(7-bromo-5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-
-5-(2-piperidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-o-
ne 56 (63 mg, yield 76.44%) as a yellow solid.
[0438] MS m/z (ESI): 515.1[M+1]
[0439] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.661 (s, 1H,
pyrrole-NH), 11.184 (s, 1H, indole-NH), 7.848.about.7.876 (dd, 1H,
--ArH), 7.794 (s, 1H, --CH.dbd.C), 7.242.about.7.240 (dd, 1H,
--ArH), 3.545.about.3.571 (t, 2H, amide N seven-membered ring
intra-CH.sub.2), 3.331.about.3.358 (t, 2H, amide N seven-membered
ring outer-CH.sub.2), 2.950.about.2.986 (t, 2H, seven-membered ring
intra-CH.sub.2C.dbd.C), 2.390.about.2.467 (m, 9H, pyrrole-CH.sub.3,
3.times.-CH.sub.2N), 2.055.about.2.084 (1, 2H, seven-membered ring
intra-CH.sub.2), 1.476.about.1.489 (m, 4H, six-membered
ring-2.times.-CH.sub.2), 1.383.about.1.393 (m, 2H, six-membered
ring-CH.sub.2).
Example 57
(Z)-2-(4-Bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-pipe-
ridin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00168##
[0441] The title compound was prepared under the same conditions as
described in step of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and 4-bromo-1,3-dihydro-indol-2-one as starting materials to obtain
(Z)-2-(4-bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-3-methyl-5-(2-pip-
eridin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
57 (68 mg, yield 91.2%) as a yellow solid.
[0442] MS m/z (ESI): 499.1[M+1]
[0443] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.682 (s, 1H,
pyrrole-NH), 11.232 (s, 1H, indole-NH), 8.634 (s, 1H, --CH.dbd.C),
7.267.about.7.287 (m, 1H, --ArH), 7.105.about.7.144 (m, 1H, --ArH),
6.988.about.7.007 (d, 1H, --ArH), 3.595.about.3.627 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.402 (t, 2H, amide N
seven-membered ring outer-CH.sub.2), 2.991.about.3.028 (t, 2H,
seven-membered ring intra-CH.sub.2C.dbd.C), 2.454.about.2.556 (m,
9H, pyrrole-CH.sub.3, 3.times.-CH.sub.2N), 2.102.about.2.131 (t,
2H, seven-membered ring intra-CH.sub.2), 1.523.about.1.550 (m, 4H,
six-membered ring-2.times.-CH.sub.2), 1.429.about.4.441 (m, 2H,
six-membered ring-CH.sub.2).
Example 58
(Z)-3-Methyl-2-(4-methyl-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-pip-
eridin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00169##
[0445] The title compound was prepared under the same conditions as
described in step of Example 32 with
3-methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyrrolo[3-
,2-c]azepine-2-carbaldehyde 32d obtained from step 4 of Example 32
and 4-methyl-1,3-dihydro-indol-2-one as starting materials to
obtain
(Z)-3-methyl-2-(4-methyl-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-pi-
peridin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
58 (46 mg, yield 70.9%) as a yellow solid.
[0446] MS m/z (ESI): 433.2[M+1]
[0447] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.703 (s, 1H,
pyrrole-NH), 10.926 (s, 1H, indole-NH), 7.567 (s, 1H, --CH.dbd.C),
7.035.about.7.074 (m, 1H, --ArH), 6.769.about.6.838 (dd, 2H,
--ArH), 3.544.about.3.576 (t, 2H, seven-membered ring
intra-CH.sub.2N), 3.331.about.3.359 (t, 2H, amide N seven-membered
ring outer-CH.sub.2), 2.926.about.2.962 (t, 2H, seven-membered ring
intra-CH.sub.2C.dbd.C), 2.591 (s, 3H, benzene-CH.sub.3),
2.383.about.2.437 (m, 9H, --CH.sub.3, pyrrole-3.times.-CH.sub.2N),
2.030.about.2.092 (t, 2H, seven-membered ring intra-CH.sub.2),
1.479.about.1.491 (m, 4H, six-membered ring-2.times.-CH.sub.2),
1.383.about.1.394 (m, 2H, six-membered ring-CH.sub.2).
Example 59
(R,Z)-2-(5-Bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-hydroxy-3-m-
orpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin--
4-one
##STR00170##
[0449] The title compound was prepared under the same conditions as
described in step 6 of Example 53 with
5-(2-hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydr-
o-pyrrolo[3,2-c]azepine-2-carbaldehyde 53f obtained from step 5 of
Example 53 and 5-bromo-1,3-dihydro-indol-2-one as starting
materials to obtain
(R,Z)-2-(5-bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-hydroxy-3--
morpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-
-4-one 59 (30 mg, yield 63%) as a yellow solid.
[0450] MS m/z (ESI): 529.1[M+1]
[0451] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.674 (s, 1H,
pyrrole-NH), 11.014 (s, 1H, indole-NH), 8.116.about.8.120 (d, 1H,
--ArH), 7.807 (s, 1H, --CH.dbd.C), 7.262.about.7.287 (dd, 1H,
--ArH), 6.826.about.6.846 (d, 1H, --ArH), 4.719.about.4.731 (d, 1H,
--OH), 3.897 (m, 1H, --CHO), 3.748.about.3.758 (dd, 1H, amide N
seven-membered ring outer-CH.sub.2), 3.570.about.3.593 (t, 4H,
morpholine-2.times.-CH.sub.2O), 3.433 (t, 2H, N seven-membered
ring-CH.sub.2), 3.159 (dd, 1H, amide N seven-membered ring
outer-CH.sub.2), 2.917.about.2.954 (t, 2H, --CH.sub.2C.dbd.C),
2.465 (s, 3H, pyrrole-CH.sub.3), 2.418.about.2.465 (m, 4H,
--CH.sub.2N, morpholine-CH.sub.2N), 2.290.about.2.314 (t, 2H,
morpholine-CH.sub.2N), 2.061.about.2.092 (m, 2H, seven-membered
ring-CH.sub.2).
Example 60
(Z)-5-(2-Diethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidene-
methyl)-3-methyl-6,7,8,9-tetrahydro-1H,5H-1,5-diaza-cyclopentacycloocten-4-
-one
##STR00171##
##STR00172## ##STR00173##
[0452] Step 1
5-(Cyclopropyl-hydroxy-methyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester
[0453] Cyclopropylmagnesium bromide (15 m, 0.5 mol/L) was cooled
down to -10.degree. C. in an ice-salt bath under an argon
atmosphere. 5-Formyl-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester 1b (1.26 g, 4.5 mmol) was
dissolved in 10 ml of tetrahydrofuran under stirring, the resulting
solution was added dropwise to the above solution while maintaining
the temperature at -10.degree. C. Upon completion of the addition,
the ice-salt bath was removed, and the reaction mixture was stirred
for 1 hour at room temperature. After thin lay chromatography
showed the disappearance of starting materials, the reaction
mixture was quenched with water, added with sulfuric acid solution
(20 ml, 10%) and stirred for 30 minutes, and extracted with ethyl
acetate (50 ml.times.3). The combined organic extracts were washed
with saturated brine (50 ml), dried over anhydrous magnesium
sulfate, filtered to remove the drying agent and concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to obtain the title compound
5-(cyclopropyl-hydroxy-methyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester 60a (576 mg, yield 39.6%) as
a white solid.
[0454] MS m/z (ESI): 322.2[M-1]
Step 2
5-(4-Bromo-but-1-enyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester
[0455]
5-(Cyclopropyl-hydroxy-methyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester 60a (323 mg, 1 mmol) was
dissolved in 4 ml of ethanol under stirring, and added with
hydrobromic acid (2.8 ml, 40%) to the solution and stirred for 30
minutes at room temperature. After thin lay chromatography showed
the disappearance of starting materials, the reaction mixture was
extracted with ethyl acetate (10 ml.times.5). The combined organic
extracts were washed with saturated brine (15 ml), dried over
anhydrous magnesium sulfate, filtered to remove the drying agent
and concentrated under reduced pressure. The residue was purified
by silica gel column chromatography to obtain the title compound
5-(4-bromo-but-1-enyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester 60b (345 mg, yield 89.5%) as a
white solid.
[0456] MS m/z (ESI): 329.4[M+1]
Step 3
5-(4-Bromo-butyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester
[0457] 5-(4-Bromo-but-1-enyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester 60b (30 mg, 0.08 mmol) was
dissolved in 3 ml of ethanol under stirring, and added with
palladium on activated carbon (6 mg, 5%) to the solution at room
temperature. The reaction mixture was stirred for 45 minutes under
a hydrogen atmosphere. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
filtered, concentrated under reduced pressure to obtain the title
compound 5-(4-bromo-butyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester 60c (21 mg, yield 70%) as a
colorless oil.
[0458] MS m/z (ESI): 388.0[M+1]
Step 4
5-(4-Diethylamino-butyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester
[0459] 5-(4-Bromo-butyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester 60c (220 mg, 0.57 mmol) was
dissolved in 5 ml of dichloromethane under stirring, and added with
N,N-diethylethylenediamine (164 .mu.l, 1.13 mmol) to the solution
and refluxed for 30 minutes in an oil bath. The reaction mixture
was concentrated to evaporate solvent and refluxed for another 1
hour. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was concentrated under
reduced pressure, purified by silica gel column chromatography to
obtain the title compound
5-(4-diethylamino-butyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester 60d (187 mg, yield 78%) as a white
solid.
[0460] MS m/z (ESI): 424.3 [M+1]
Step 5
5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-4,5,6,7,8,9-hexahydro-1H-1,5-diaza-
-cyclopentacyclooctene-2-carbaldehyde
[0461] Trimethyl aluminum (489 .mu.l, 2 mol/L) was dissolved in 3
ml of toluene under stirring, and added with the solution of
5-(4-diethylamino-butyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic acid
2-tert-butyl ester 4-ethyl ester 60d (345 mg, 0.82 mmol) in 6 ml of
toluene to the above solution at room temperature. Upon completion
of the addition, the reaction mixture was stirred for 30 minutes at
room temperature, heated to reflux for 2 hours in an oil bath,
added with another trimethyl aluminum (900 .mu.l, 2 mol/L) and
refluxed for another 7 hours. After thin lay chromatography showed
the disappearance of starting materials, the reaction mixture was
quenched with water, added with hydrochloric acid solution (1 ml, 2
mol/L) and stirred for 30 minutes at room temperature. The mixture
was adjusted to about pH 10 with aqueous sodium hydroxide solution
(10%) and extracted with ethyl acetate (25 ml.times.3). The
combined organic extracts were washed with saturated brine (25 ml),
dried over anhydrous magnesium sulfate, filtered to remove the
drying agent and concentrated under reduced pressure. The residue
was purified by silica gel column chromatography to obtain the
title compound
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-4,5,6,7,8,9-hexahydro-1H-1,5-diaz-
a-cyclopentacyclooctene-2-carbaldehyde 60e (60 mg, yield 26.7%) as
a white solid.
[0462] MS m/z (ESI): 278.2[M+1]
Step 6
(Z)-5-(2-Diethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidene-
methyl)-3-methyl-6,7,8,9-tetrahydro-1H,5H-1,5-diaza-cyclopentacycloocten-4-
-one
[0463]
5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-4,5,6,7,8,9-hexahydro-1H-1,-
5-diaza-cyclopentacyclooctene-2-carbaldehyde 60e (20 mg, 0.066
mmol) was dissolved in 1 ml of ethanol under stirring, and added
with 5-fluoro-1,3-dihydro-indol-2-one (9.9 mg, 0.066 mmol) to the
solution at room temperature. The reaction mixture was stirred in
dark until dissolved, added with 0.1 ml of piperidine, and heated
to reflux for 2 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature, filtered to obtain the
title compound
(Z)-5-(2-diethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-yliden-
emethyl)-3-methyl-6,7,8,9-tetrahydro-1H,5H-1,5-diaza-cyclopentacycloocten--
4-one 60 (14 mg, yield 48.8%) as a yellow solid.
[0464] MS m/z (ESI): 439.2 [M+1]
[0465] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.462 (s, 1H,
pyrrole-NH), 10.874 (s, 1H, indole-NH), 7.739.about.7.758 (d, 1H,
--ArH), 7.715 (s, 1H, --CH.dbd.C), 6.832.about.6.933 (m, 2H,
--ArH), 3.406 (m, 4H, 2.times.-CH.sub.2NCO), 2.874 (t, 2H,
--CH.sub.2C.dbd.C), 2.597.about.2.630 (t, 2H, --CH.sub.2N),
2.486.about.2.538 (q, 4H, ethyl 2.times.-CH.sub.2N), 2.322 (s, 3H,
pyrrole-CH.sub.3), 1.733 (m, 4H, eight-membered ring
intra-2.times.-CH.sub.2), 0.963.about.0.968 (t, 6H,
2.times.-CH.sub.3).
Example 61
(Z)-2-((5-Bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-diethylamino-
-ethyl)-3-methyl-6,7,8,9-tetrahydro-1H,5H-1,5-diaza-cyclopentacycloocten-4-
-one
##STR00174##
[0467] The title compound was prepared under the same conditions as
described in step 6 of Example 60 with
5-(2-diethylamino-ethyl)-3-methyl-4-oxo-4,5,6,7,8,9-hexahydro-1H-1,5-diaz-
a-cyclopentacyclooctene-2-carbaldehyde 60e obtained from step 5 of
Example 60 and 5-bromo-1,3-dihydro-indol-2-one as starting
materials to obtain
(Z)-2-((5-bromo-2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-5-(2-diethylamin-
o-ethyl)-3-methyl-6,7,8,9-tetrahydro-1H,5H-1,5-diaza-cyclopentacycloocten--
4-one 61 (16 mg, yield 68%) as a yellow solid.
[0468] MS m/z (ESI): 499.2[M+1]
[0469] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.660 (s, 1H,
pyrrole-NH), 11.008 (s, 1H, indole-NH), 8.113.about.8.117 (d, 1H,
--ArH), 7.803 (s, 1H, --CH.dbd.C), 7.260.about.7.286 (dd, 1H,
--ArH), 6.825.about.6.845 (d, 1H, --ArH), 3.406 (m, 41-1,
2.times.-CH.sub.2NCO), 2.874 (t, 2H, --CH.sub.2C.dbd.C),
2.597.about.2.630 (t, 2H, --CH.sub.2N), 2.486.about.2.538 (q, 4H,
ethyl 2.times.-CH.sub.2N), 2.322 (s, 3H, pyrrole-CH.sub.3), 1.733
(m, 4H, eight-membered ring intra-2.times.-CH.sub.2),
0.963.about.0.968 (t, 6H, 2.times.-CH.sub.3).
Example 62
(Z)-5-(2-Ethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-ylideneme-
thyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
##STR00175##
##STR00176## ##STR00177##
[0470] Step 1
5-[2-(2-Acetylamino-ethylcarbamoyl)-ethyl]-3-methyl-1H-pyrrole-2,4-dicarbo-
xylic acid 2-tert-butyl ester 4-ethyl ester
[0471] 5-(2-Carboxy-ethyl)-3-methyl-1H-pyrrole-2,4-dicarboxylic
acid 2-tert-butyl ester 4-ethyl ester le (9.85 g, 30.3 mmol)
obtained from step 4 of Example 1 was dissolved in acetonitrile (50
ml) under stirring, and added with 1-hydroxy-1H-benzotriazole (8.2
g, 60.6 mmol) and N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide
(11.6 g, 60.6 mmol) to the solution at room temperature. Upon
completion of the addition, the reaction mixture was stirred at
room temperature overnight. After thin lay chromatography showed
the disappearance of starting materials, the reaction mixture was
concentrated under reduced pressure, added with water (200 ml), and
exacted with ethyl acetate (200 ml.times.4). The combined organic
extracts were washed with saturated brine (100 ml), dried over
anhydrous magnesium sulfate, filtered to remove the drying agent
and concentrated under reduced pressure. The residue was purified
by silica gel column chromatography to obtain the title compound
5-[2-(2-acetylamino-ethylcarbamoyl)-ethyl]-3-methyl-1H-pyrrole-2,4-dicarb-
oxylic acid 2-tert-butyl ester 4-ethyl ester 62a (8 g, yield 65%)
as a white solid.
[0472] MS m/z (ESI): 410.1[M+1]
Step 2
2-[2-(2-Acetylamino-ethylcarbamoyl)-ethyl]-4-methyl-1H-pyrrole-3-carboxyli-
c acid ethyl ester
[0473]
5-[2-(2-Acetylamino-ethylcarbamoyl)-ethyl]-3-methyl-1H-pyrrole-2,4--
dicarboxylic acid 2-tert-butyl ester 4-ethyl ester 62a (818 mg, 2
mmol) was dissolved in 5 ml of ethanol under stirring, and added
with hydrochloric acid solution (5 ml, 12 mol/L) to the solution at
room temperature. Upon completion of the addition, the reaction
mixture was heated at 60.degree. C. in an oil bath for 2 hours.
After thin lay chromatography showed the disappearance of starting
materials, the reaction mixture was concentrated under reduced
pressure to evaporate ethanol, adjusted to about pH 12 with aqueous
sodium hydroxide solution (10%) and extracted with ethyl acetate
(20 ml.times.3). The combined organic extracts were dried over
anhydrous magnesium sulfate, filtered to remove the drying agent
and concentrated under reduced pressure to obtain the title
compound
2-[2-(2-acetylamino-ethylcarbamoyl)-ethyl]-4-methyl-1H-pyrrole-3-carboxyl-
ic acid ethyl ester 62b (600 mg, yield 97%) as a white solid.
[0474] MS m/z (ESI): 310.1[M+1]
Step 3
2-[3-(2-Ethylamino-ethylamino)-propyl]-4-methyl-1H-pyrrole-3-carboxylic
acid ethyl ester
[0475]
2-[2-(2-Acetylamino-ethylcarbamoyl)-ethyl]-4-methyl-1H-pyrrole-3-ca-
rboxylic acid ethyl ester 62b (600 mg, 1.94 mmol) was dissolved in
4 ml of tetrahydrofuran under stirring, and added with a solution
of borane in tetrahydrofuran (7.79 ml, 1 mol/L, 7.79 mmol) to the
solution in an ice-water bath under an argon atmosphere. Upon
completion of the addition, the reaction mixture was heated to
reflux for 2 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
quenched with hydrochloric acid, stirred for 30 minutes at room
temperature, adjusted to about pH 12 with aqueous sodium hydroxide
solution (10%) and extracted with ethyl acetate (30 ml.times.3).
The combined organic extracts were washed with saturated brine (30
ml), dried over anhydrous magnesium sulfate, filtered to remove the
drying agent and concentrated under reduced pressure. The residue
was purified by silica gel column chromatography to obtain the
title compound
2-[3-(2-ethylamino-ethylamino)-propyl]-4-methyl-1H-pyrrole-3-carboxylic
acid ethyl ester 62c (170 mg, yield 31%) as a white solid.
[0476] MS m/z (ESI): 282.2[M+1]
Step 4
5-(2-Ethylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-
-4-one
[0477]
2-[3-(2-Ethylamino-ethylamino)-propyl]-4-methyl-1H-pyrrole-3-carbox-
ylic acid ethyl ester 62c (350 mg, 1.245 mmol) and trimethyl
aluminum (1.25 ml, 2.49 mmol) were dissolved in 75 ml of toluene
under stirring, the resulting mixture was heated to reflux at
140.degree. C. in an oil bath for 24 hours. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was concentrated under reduced pressure, adjusted
to about pH 3 with hydrochloric acid (6 mol/L) and stirred for 30
minutes. The mixture was adjusted to pH 14 with aqueous sodium
hydroxide solution (12 mol/L) and extracted with dichloromethane
(100 ml.times.4). The combined organic extracts were washed with
saturated brine (150 ml), dried over anhydrous magnesium sulfate,
filtered to remove the drying agent and concentrated under reduced
pressure to obtain the title compound
5-(2-ethylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepi-
n-4-one 62d (450 mg) as a yellow oil to be used directly in the
next step.
[0478] MS m/z (ESI): 236.0[M+1]
Step 5
Ethyl-[2-(3-methyl-4-oxo-4,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-5-yl)--
ethyl]-carbamic acid tert-butylester
[0479]
5-(2-Ethylamino-ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c-
]azepin-4-one 62d (450 mg, 1.91 mmol) was dissolved in 20 ml of
dichloromethane under stirring, and added with di-tert-butyl
dicarbonate (834 mg, 3.83 mmol), potassium carbonate (528 mg, 3.83
mmol) and tetrahydrofuran (30 ml) to the solution. Upon completion
of the addition, the reaction mixture was stirred at room
temperature overnight. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was added
with water (50 ml), and exacted with ethyl acetate (50 ml.times.3).
The combined organic extracts were washed with saturated brine (50
ml), dried over anhydrous magnesium sulfate, filtered to remove the
drying agent and concentrated under reduced pressure. The residue
was purified by silica gel column chromatography to obtain the
title compound
ethyl-[2-(3-methyl-4-oxo-4,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-5-yl)-
-ethyl]-carbamic acid tert-butylester 62e (40 mg, yield 6%) as a
yellow solid.
[0480] MS m/z (ESI): 336.2[M+1]
Step 6
Ethyl-[2-(2-formyl-3-methyl-4-oxo-4,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azep-
in-5-yl)-ethyl]-carbamic acid tert-butyl ester
[0481]
Ethyl-[2-(3-methyl-4-oxo-4,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-
-5-yl)-ethyl]-carbamic acid tert-butylester 62e (40 mg, 0.119 mmol)
was dissolved in 25 ml of dichloromethane under stirring, and added
with (chloromethylene)dimethylammoniumchloride (15.92 mg, 0.12
mmol) to the solution. Upon completion of the addition, the
reaction mixture was stirred at room temperature for 10 minutes.
After thin lay chromatography showed the disappearance of starting
materials, the reaction mixture was quenched with aqueous sodium
hydroxide solution (12 mol/L), stirred for another 15 minutes at
room temperature and extracted with dichloromethane (20
ml.times.3). The combined organic extracts were washed with
saturated brine (20 ml), dried over anhydrous magnesium sulfate,
filtered to remove the drying agent and concentrated under reduced
pressure to obtain the title compound
ethyl-[2-(2-formyl-3-methyl-4-oxo-4,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]aze-
pin-5-yl)-ethyl]-carbamic acid tert-butyl ester 62f (54 mg) as a
yellow solid to be used directly in the next step.
[0482] MS m/z (ESI): 364.1[M+1]
Step 7
5-(2-Ethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-c]-
azepine-2-carbaldehyde
[0483]
Ethyl-[2-(2-formyl-3-methyl-4-oxo-4,6,7,8-tetrahydro-1H-pyrrolo[3,2-
-c]azepin-5-yl)-ethyl]-carbamic acid tert-butyl ester 62f (43 mg,
0.118 mmol) was dissolved in 10 ml of dichloromethane under
stirring, and added with trifluoroacetic acid (272 .mu.A, 3.55
mmol) to the solution and stirred for 15 minutes in an ice-water
bath. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was concentrated under
reduced pressure to obtain the title compound
5-(2-ethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo[3,2-c-
]azepine-2-carbaldehyde 62j to be used directly in the next
step.
[0484] MS m/z (ESI): 264.1[M+1]
Step 8
(Z)-5-(2-Ethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-ylideneme-
thyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one
[0485]
5-(2-Ethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrrolo-
[3,2-c]azepine-2-carbaldehyde 62j (45 mg, 0.118 mmol) was dissolved
in 5 ml of ethanol under stirring, and added with
5-fluoro-1,3-dihydro-indol-2-one (16 mg, 0.106 mmol) and piperidine
(0.15 ml, 1.49 mmol) to the solution. Upon completion of the
addition, the reaction mixture was refluxed at 90.degree. C. in an
oil bath for 1 hour. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
concentrated under reduced pressure, added with a little ethanol,
and filtered to obtain the title compound
(Z)-5-(2-ethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidenem-
ethyl)-3-methyl-5,6,7,8-tetrahydro-1H-pyrrolo[3,2-c]azepin-4-one 62
(34 mg, yield 76%) as a yellow solid.
[0486] MS m/z (ESI): 397.1[M+1]
[0487] .sup.1H NMR (400 MHz, DMSO-d6) 8 (s, 1H, pyrrole-NH), 10.920
(s, 1H, indole-NH), 7.781.about.7.788 (d, 1H, --ArH), 7.759 (s, 1H,
--CH.dbd.C), 6.925.about.6.976 (td, 1H, --ArH), 6.845.about.6.877
(dd, 1H, --ArH), 3.637.about.3.688 (t, 2H, N seven-membered
ring-CH.sub.2), 3.371.about.3.398 (t, 2H, amide N seven-membered
ring outer-CH.sub.2), 2.959.about.3.026 (m, 4H, --CH.sub.2C.dbd.C,
--CH.sub.2N), 2.864.about.2.918 (q, 2H, ethyl-CH.sub.2), 2.488 (s,
3H, pyrrole-CH.sub.3), 2.056.about.2.083 (m, 2H, seven-membered
ring-CH.sub.2), 1.136.about.1.172 (t, 3H, ethyl-CH.sub.3)
Example 63
(Z)-5-(2-Diethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-ylidene-
methyl)-methyl-3a,5,6,7,8,8a-hexahydro-1H-pyrrolo[3,2-c]azepin-4-one
malate
##STR00178##
[0489]
(Z)-5-(2-Diethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3--
ylidenemethyl)-3-methyl-3a,5,6,7,8,8a-hexahydro-1H-pyrrolo[3,2-c]azepin-4--
one 1 (2.01 g, 4.75 mmol) obtained from Example 1 was dissolved in
279 ml of methanol under stirring, and added with
2-hydroxy-succinic acid (0.953 g, 7.11 mmol) to the solution in one
portion. The orange solution was concentrated under reduced
pressure, added with 45 ml of acetonitrile, heated to reflux for 30
minutes in an oil bath. Then the oil bath was removed, and the
reaction mixture was naturally cooled down to room temperature,
filtered and dried to obtain the title compound
(Z)-5-(2-diethylamino-ethyl)-2-(5-fluoro-2-oxo-1,2-dihydro-indol-3-yliden-
emethyl)-3-methyl-3a,5,6,7,8,8a-hexahydro-1H-pyrrolo[3,2-c]azepin-4-one
malate 63 (2.02 g, yield 76.2%) as an orange solid.
[0490] MS m/z (ESI): 425.1[M+1]
[0491] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.742 (s, 1H,
pyrrole-NH), 10.925 (s, 1H, indole-NH), 7.784.about.7.790 (dd, 1H,
--ArH), 7.755 (s, 1H, --CH.dbd.C), 6.922.about.6.951 (m, 1H,
--ArH), 6.840.about.6.873 (m, 1H, --ArH), 3.631.about.3.665 (t, 2H,
seven-membered ring intra-CH.sub.2N), 3.374.about.3.401 (t, 2H,
amide N seven-membered ring outer-CH.sub.2), 2.911.about.2.958 (t,
2H, seven-membered ring intra-CH.sub.2C.dbd.C), 2.536.about.2.575
(m, 6H, 3.times.-CH.sub.2N), 2.471 (s, 3H, pyrrole-CH.sub.3),
2.053.about.2.079 (m, 2H, seven-membered ring intra-CH.sub.2),
1.137 (t, 6H, 2.times.-CHO.
Example 64
(Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-3-me-
thyl-5-(2-morpholinoethyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-on-
e
##STR00179##
##STR00180##
[0492] Step 1
5-Chlorosulfonyl-indol-2-one
[0493] 1,3-Dihydro-indol-2-one 64a (13.3 g, 100 mmol) was added
slowly with chlorosulfuric acid (26.6 ml, 400 mmol) in an ice-water
bath. Upon completion of the addition, the reaction mixture was
stirred for 1 hour in an ice-water bath, for another 1 hour at room
temperature, and heated to 68.degree. C. for 1 hour. The reaction
mixture was cooled down to room temperature, added slowly with
water (400 ml), stirred and yellow precipitates were formed. After
standing for 1 hour at room temperature, the filter cake was washed
with water (20 ml.times.4) and dried to obtain the title compound
5-chlorosulfonyl-indol-2-one 64b (15.0 g, yield 65%) as a yellow
solid. [0494] Reference: Acta Pharmacol Sin; 2007, 28(1),
140-152.
Step 2
5-(2,6-Dichloro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one
[0495] Sodium monohydrogen phosphate dodecahydrate (142 g, 1.0 mol)
and sodium sulfite (252 g, 2.0 mol) were dissolved in 2 L of water
at room temperature, heated to 30.degree. C., and added with
2-oxo-2,3-dihydro-1H-indole-5-sulfonyl chloride 64b (232 g, 1.0
mol). Upon completion of the addition, the reaction mixture was
stirred at 60.degree. C. for 16 hours. A solution of
2,6-dichlorobenzyl bromide (240 g, 1.0 mol) in acetone (1.8 L) was
added to the above solution, stirred at 60.degree. C. for 1 hour,
added with another acetone (200 ml) and stirred for another 2
hours. The reaction mixture was quenched with water (5 L), stirred
at room temperature for 1 hour and filtered. The filter cake was
washed with water (1 L) and acetone (1 L), and dried in vacuo to
obtain the title compound
5-(2,6-dichloro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 64c
(314 g, yield 88%) as a white solid.
[0496] MS m/z (ESI): 357.3 [M+1]
[0497] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 10.90 (s, 1H,
indole-NH), 7.56 (m, 4H, --ArH), 7.43 (m, 1H, --ArH), 6.99 (d, 1H,
--ArH), 4.59 (s, 2H, --ArCH.sub.2) [0498] Reference: Organic
Process Research & Development; 2003, 7, 313-317.
Step 3
(Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-3-me-
thyl-5-(2-morpholinoethyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-on-
e
[0499]
3-Methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyr-
rolo[3,2-c]azepine-2-carbaldehyde 10c (100 mg, 0.325 mmol) and
5-(2,6-dichloro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 64c
(104 mg, 0.293 mmol) were dissolved in 3 ml of ethanol, and added
with 52 .mu.l of piperidine to the solution at room temperature.
Upon completion of the addition, the reaction mixture was heated to
reflux for 2 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature and filtered. The filter
cake was washed with anhydrous ethanol (3 ml.times.2) and dried to
obtain the title compound
(Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-3-m-
ethyl-5-(2-morpholinoethyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-o-
ne 64 (166 mg, yield 88%) as an orange solid.
[0500] MS m/z (ESI): 643.3[M+1]
[0501] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.63 (s, 1H,
pyrrole-NH), 11.42 (s, 1H, indole-NH), 8.28 (s, 1H, --ArH), 7.90
(s, 1H, --ArH), 7.51 (m, 3H, --ArH), 7.42 (s, 1H, --CH.dbd.C), 7.06
(m, 1H, --ArH), 4.88 (s, 2H, --ArCH.sub.2), 3.58-2.08 (m, 18H,
aliphatic H), 2.44 (s, 3H, pyrrole-CH.sub.3)
Example 65
(Z)-2-[5-(4-Fluoro-phenylmethanesulfonyl)-2-oxo-1,2-dihydro-indol-3-yliden-
emethyl]-3-methyl-5-(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrol-
o[3,2-c]azepin-4-one
##STR00181##
##STR00182##
[0502] Step 1
5-(4-Fluoro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one
[0503] Sodium monohydrogen phosphate dodecahydrate (3.58 g, 10
mmol) and sodium sulfite (2.52 g, 20 mmol) were dissolved in 20 ml
of water, heated to 30.degree. C., and added with
2-oxo-2,3-dihydro-1H-indole-5-sulfonyl chloride 64b (2.32 g, 10
mmol). Upon completion of the addition, the reaction mixture was
stirred at 60.degree. C. for 16 hours. A solution of 5-fluorobenzyl
bromide (1.9 g, 10 mmol) in acetone (18 ml) was added slowly to the
above solution and stirred at 60.degree. C. for 2 hours. The
reaction mixture was quenched with water (50 ml) and lots of
precipitates were formed. The mixture was stirred at room
temperature for another 1 hour, filtered, washed with the solvent
mixture (20 ml, V:V=1:1) of water and acetone, and dried in vacuo
to obtain the title compound
5-(4-fluoro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 65a (1.8
g, yield 59%) as a light yellow solid.
[0504] MS m/z (ESI): 304.1[M-1]
[0505] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 10.85 (s, 1H,
indole-NH), 7.42 (m, 2H, --ArH), 7.21 (m, 3H, --ArH), 6.95 (d, 1H,
--ArH), 4.59 (s, 2H, --ArCH.sub.2) [0506] Reference: Organic
Process Research & Development; 2003, 7, 313-317.
Step 2
2-[5-(4-Fluoro-phenylmethanesulfonyl)-2-oxo-1,2-dihydro-indol-3-ylidenemet-
hyl]-3-methyl-5-(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrrolo[3,-
2-c]azepin-4-one
[0507]
3-Methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-py-
rrolo[3,2-c]azepine-2-carbaldehyde 28c (60.2 mg, 0.21 mmol) and
5-(4-fluoro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 65a
(57.6 mg, 0.19 mmol) were dissolved in 2 ml of ethanol, and added
with 50 .mu.l of piperidine to the solution at room temperature.
Upon completion of the addition, the reaction mixture was heated to
reflux for 3 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature, and filtered. The filter
cake was washed with anhydrous ethanol (3 ml.times.2) and dried to
obtain the title compound
(Z)-2-[5-(4-fluoro-phenylmethanesulfonyl)-2-oxo-1,2-dihydro-indol-3-ylide-
nemethyl]-3-methyl-5-(2-pyrrolidin-1-yl-ethyl)-5,6,7,8-tetrahydro-1H-pyrro-
lo[3,2-c]azepin-4-one 65 (90 mg, yield 81.8%) as an orange
solid.
[0508] MS m/z (ESI): 577.3[M+1]
[0509] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 12.133 (s, 1H,
indole-NH), 8.99 (s, 1H, --ArH), 8.61 (s, 1H, --ArH), 7.88 (s, 1H,
--CH.dbd.C), 8.14-7.76 (m, 4H, --ArH), 7.75 (d, 1H, --ArH), 5.39
(s, 2H, --ArCH.sub.2), 4.33 (m, 2H, --NCH.sub.2), 4.11.about.3.25
(m, 12H, aliphatic H), 3.70 (m, 2H, --NCH.sub.2), 2.81 (m, 2H,
--NCH.sub.2), 2.46 (s, 3H, pyrrole-CH.sub.3).
Example 66
(Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-3-me-
thyl-5-(2-(pyrrolidin-1-yl)ethyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4-
(1H)-one
##STR00183##
[0511]
3-methyl-4-oxo-5-(2-pyrrolidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-py-
rrolo[3,2-c]azepine-2-carbaldehyde 28c (75 mg, 0.262 mmol) and
5-(2,6-dichloro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 64c
(84 mg, 0.236 mmol) were dissolved in 2.5 ml of ethanol, and added
with 42 .mu.l of piperidine to the solution at room temperature.
Upon completion of the addition, the reaction mixture was heated to
reflux for 3 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature, and filtered. The filter
cake was washed with anhydrous ethanol (3 ml.times.2) and dried to
obtain the title compound
(Z)-2-.beta.5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-
-3-methyl-5-(2-(pyrrolidin-1-yl)ethyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]aze-
pin-4(1H)-one 66 (117 mg, yield 79.6%) as an orange solid.
[0512] MS m/z (ESI): 627.3[M+1]
[0513] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 12.18 (s, 1H,
indole-NH), 9.04 (s, 1H, --ArH), 8.66 (s, 1H, --ArH), 8.25 (s, 1H,
--CH.dbd.C), 8.23-8.16 (m, 3H, --ArH), 7.99 (d, 1H, --ArH), 5.64
(s, 2H, --ArCH.sub.2), 4.34 (m, 2H, --NCH.sub.2), 4.12-3.26 (m,
12H, aliphatic H), 3.70 (m, 2H, --NCH.sub.2), 2.81 (m, 2H,
--NCH.sub.2), 2.46 (s, 3H, pyrrole-CH.sub.3).
Example 67
(Z)-2-((5-(4-fluorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-3-methyl-
-5-(2-(piperidin-1-yl)ethyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)--
one
##STR00184##
[0515]
3-Methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyr-
rolo[3,2-c]azepine-2-carbaldehyde 32d (57.6 mg, 0.21 mmol) and
5-(4-fluoro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 65a
(57.6 mg, 0.19 mmol) were dissolved in 2 ml of ethanol, and added
with 42 .mu.l of piperidine to the solution at room temperature.
Upon completion of the addition, the reaction mixture was heated to
reflux for 3 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature, and filtered. The filter
cake was washed with anhydrous ethanol (3 ml.times.2) and dried to
obtain the title compound
(Z)-2-.beta.5-(4-fluorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-3-m-
ethyl-5-(2-(piperidin-1-yl)ethyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4-
(1H)-one 67 (84 mg, yield 76%) as an orange solid.
[0516] MS m/z (ESI): 591.3[M+1]
[0517] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.61 (s, 1H,
pyrrole-NH), 11.39 (s, 1H, indole-NH), 8.24 (s, 1H, --HCO), 7.86
(s, 1H, --ArH), 7.17 (s, 1H, --CH.dbd.C), 7.39-7.13 (m, 4H, --ArH),
6.99 (d, 1H, --ArH), 4.64 (s, 2H, --ArCH.sub.2), 3.58 (m, 2H,
--NCH.sub.2), 3.29-2.40 (m, 11H, aliphatic H), 2.97 (m, 2H,
--NCH.sub.2), 2.08 (m, 2H, --NCH.sub.2), 1.49 (s, 3H,
pyrrole-CH.sub.3).
Example 68
(Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-3-me-
thyl-5-(2-(piperidin-1-yl)ethyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(-
1H)-one
##STR00185##
[0519]
3-Methyl-4-oxo-5-(2-piperidin-1-yl-ethyl)-1,4,5,6,7,8-hexahydro-pyr-
rolo[3,2-c]azepine-2-carbaldehyde 32d (57.6 mg, 0.21 mmol) and
5-(2,6-dichloro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 64c
(67.2 mg, 0.19 mmol) were dissolved in 2.5 ml of ethanol, and added
with 42 .mu.l of piperidine to the solution at room temperature.
Upon completion of the addition, the reaction mixture was heated to
reflux for 3 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature and filtered. The filter
cake was washed with anhydrous ethanol (3 ml.times.2), and dried to
obtain the title compound
(Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-3-m-
ethyl-5-(2-(piperidin-1-yl)ethyl)-5,6,7,8-tetrahydro
pyrrolo[3,2-c]azepin-4(1H)-one 68 (93 mg, yield 77.5%) as an orange
solid.
[0520] MS m/z (ESI): 641.2[M+1]
[0521] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 8.79 (s, 1H, --ArH),
7.90 (s, 1H, --CH.dbd.C), 7.51-7.41 (m, 3H, --ArH), 7.05 (d, 1H,
--ArH), 4.89 (m, 2H, --ArCH.sub.2), 3.57 (m, 2H, --NCH.sub.2), 3.35
(m, 2H, --NCH.sub.2), 2.97 (m, 2H, --NCH.sub.2), 3.31-2.39 (m, 15H,
aliphatic H), 2.07 (m, 2H, --NCH.sub.2).
Example 69
(Z)-2-((5-(4-fluorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-3-methyl-
-5-(2-morpholinoethyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
##STR00186##
[0523]
3-Methyl-5-(2-morpholin-4-yl-ethyl)-4-oxo-1,4,5,6,7,8-hexahydro-pyr-
rolo[3,2-c]azepine-2-carbaldehyde 10c (80 mg, 0.26 mmol) and
5-(4-fluoro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 65a (72
mg, 0.24 mmol) were dissolved in 2.5 ml of ethanol, and added with
42 .mu.l of piperidine the solution at room temperature. Upon
completion of the addition, the reaction mixture was heated to
reflux for 3 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature, and filtered. The filter
cake was washed with anhydrous ethanol (3 ml.times.2) and dried to
obtain the title compound
(Z)-2-((5-(4-fluorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-3-methy-
l-5-(2-morpholinoethyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
69 (120 mg, yield 85.7%) as an orange solid.
[0524] MS m/z (ESI): 593.5[M+1]
[0525] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.61 (s, 1H,
pyrrole-NH), 11.39 (s, 1H, indole-NH), 8.24 (s, 1H, --ArH), 7.86
(s, 1H, --ArH), 7.40 (m, 1H, --ArH), 7.24 (m, 3H, 3.times.-ArH),
7.21 (s, 1H, --CH.dbd.C), 7.01 (d, 1H, --ArH), 4.64 (s, 2H,
--ArCH.sub.2), 3.58-2.07 (m, 18H, aliphatic H), 2.44 (s, 3H,
pyrrole-CH.sub.3).
Example 70
(Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-5-(2-
-(diethylamino)ethyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H-
)-one
##STR00187##
[0527]
5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrro-
lo[3,2-c]azepine-2-carbaldehyde 1j (60 mg, 0.21 mmol) and
5-(2,6-dichloro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 64c
(67 mg, 0.19 mmol) were dissolved in 3 ml of ethanol, and added
with 52 .mu.l of piperidine the solution at room temperature. Upon
completion of the addition, the reaction mixture was heated to
reflux for 2 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature, and filtered. The filter
cake was washed with anhydrous ethanol (3 ml.times.2) and dried to
obtain the title compound
(Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-5-(-
2-(diethylamino)ethyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1-
H)-one 70 (95 mg, yield 79%) as an orange solid.
[0528] MS m/z (ESI): 629.3[M+1]
[0529] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.62 (s, 1H,
pyrrole-NH), 11.42 (s, 1H, indole-NH), 8.29 (s, 1H, --ArH), 7.90
(s, 1H, --ArH), 7.49 (s, 1H, --CH.dbd.C), 7.52 (m, 2H,
--NCH.sub.2), 7.06 (s, 1H, --ArH), 4.89 (s, 2H, --ArCH.sub.2),
3.52-2.07 (m, 14H, aliphatic H), 2.50 (s, 3H, pyrrole-CH.sub.3),
1.00 (m, 6H, 2.times.-CH.sub.3).
Example 71
(Z)-2-((5-(4-fluorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-5-(2-(di-
ethylamino)ethyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-on-
e
##STR00188##
[0531]
5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrro-
lo[3,2-c]azepine-2-carbaldehyde 1j (60 mg, 0.21 mmol) and
5-(4-fluoro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 65a (58
mg, 0.19 mmol) were dissolved in 2 ml of ethanol, and added with 34
.mu.l of piperidine the solution at room temperature. Upon
completion of the addition, the reaction mixture was heated to
reflux for 3 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature, and filtered. The filter
cake was washed with anhydrous ethanol (3 ml.times.2) and dried to
obtain the title compound
(Z)-2-((5-(4-fluorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-5-(2-(d-
iethylamino)ethyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-o-
ne 71 (73 mg, yield 66%) as an orange solid.
[0532] MS m/z (ESI): 579.3[M+1]
[0533] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.59 (s, 1H,
pyrrole-NH), 11.39 (s, 1H, indole-NH), 8.24 (s, 1H, --ArH), 7.86
(s, 1H, --ArH), 7.21 (s, 1H, --CH.dbd.C), 7.24-7.13 (m, 3H, --ArH),
7.01 (d, 1H, --ArH), 4.64 (s, 2H, --ArCH.sub.2), 3.52-2.06 (m, 14H,
aliphatic H), 2.50 (s, 3H, pyrrole-CH.sub.3), 1.00 (m, 6H,
2.times.-CH.sub.3).
Example 72
(R,Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-5--
(2-hydroxy-3-morpholinopropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]az-
epin-4(1H)-one
##STR00189##
[0535]
5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-he-
xahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 53f (80 mg, 0.24 mmol)
and 5-(2,6-dichloro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one
64c (75 mg, 0.21 mmol) were dissolved in 3 ml of ethanol, and added
with 30 .mu.l of piperidine to the solution at room temperature.
Upon completion of the addition, after stirring for 4 minutes, the
reaction mixture was reacted at 120.degree. C. for 4 minutes under
microwave and became orange turbid solution. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was naturally cooled down to room temperature, and
filtered. The filter cake was washed with anhydrous ethanol (2
ml.times.2) and dried to obtain the title compound
(R,Z)-2-((5-(2,6-dichlorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-5-
-(2-hydroxy-3-morpholinopropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]a-
zepin-4(1H)-one 72 (120 mg, yield 85%) as a yellow solid.
[0536] MS m/z (ESI): 673.2[M+1]
[0537] .sup.1HNMR (400 MI-1z, DMSO-d6) .delta. 13.63 (s, 1H,
pyrrole-NH), 11.42 (s, 1H, indole-NH), 8.29 (d, 1H, --ArH), 7.90
(s, 1H, --ArH), 7.52 (m, 4H, --ArH), 7.42 (s, 1H, --CH.dbd.C), 7.06
(d, 1H, --ArH), 4.89 (s, 2H, --ArCH.sub.2), 4.74 (m, 1H, --CHO),
3.92-2.33 (m, 18H, aliphatic H), 2.44 (s, 3H,
pyrrole-CH.sub.3).
Example 73
(R,Z)-2-((5-(4-fluorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-5-(2-h-
ydroxy-3-morpholinopropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-
-4(1H)-one
##STR00190##
[0539]
5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-he-
xahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 53f (80 mg, 0.24 mmol)
and 5-(4-fluoro-phenylmethanesulfonyl)-1,3-dihydro-indol-2-one 65a
(64 mg, 0.21 mmol) were dissolved in 3 ml of ethanol, and added
with 30 .mu.l of piperidine to the solution at room temperature.
Upon completion of the addition, after stirring for 4 minutes, the
reaction mixture was reacted at 120.degree. C. for 4 minutes under
microwave and became orange turbid solution. After thin lay
chromatography showed the disappearance of starting materials, the
reaction mixture was naturally cooled down to room temperature, and
filtered. The filter cake was washed with anhydrous ethanol (2
ml.times.2) and dried to obtain the title compound
(R,Z)-2-((5-(4-fluorobenzylsulfonyl)-2-oxoindolin-3-ylidene)methyl)-5-(2--
hydroxy-3-morpholinopropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepi-
n-4(1H)-one 73 (100 mg, yield 76%) as a yellow solid.
[0540] MS m/z (ESI): 621.3[M-1]
[0541] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.61 (s, 1H,
pyrrole-NH), 11.38 (s, 1H, indole-NH), 8.24 (d, 1H, --ArH), 7.86
(s, 1H, --ArH), 7.40 (m, 1H, --ArH), 7.24 (m, 1H, --ArH), 7.17 (m,
1H, --ArH), 7.13 (s, 1H, --CH.dbd.C), 7.01 (d, 1H, --ArH), 4.74 (m,
1H, --CHO), 4.64 (s, 2H, --ArCH.sub.2), 3.92-2.32 (m, 18H,
aliphatic H), 2.44 (s, 3H, pyrrole-CH.sub.3).
Example 74
(Z)-5-(2-(diethylamino)ethyl)-2-((4-(2,3-difluorophenyl)-5-fluoro-2-oxoind-
olin-3-ylidene)methyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1-
H)-one
##STR00191##
##STR00192##
[0542] Step 1
1-Iodo-2-methyl-3-nitro-benzene
[0543] 2-Methyl-3-nitro-phenylamine 74a (21.28 g, 0.14 mol) was
dissolved in 70 ml of concentrated hydrochloric acid in an
ice-water bath, added with water (40 ml), stirred at
0.about.5.degree. C. and yellow-green precipitates were formed. The
reaction mixture was added dropwise with nitric acid solution (40
ml, 3.6 M), stirred for 15 minutes and filtered. The filtrate was
added dropwise to potassium iodide solution (280 ml, 5.25 M) at
0.about.5.degree. C. Upon completion of the addition, the reaction
mixture was stirred for 1 hour. After thin lay chromatography
showed the disappearance of starting materials, the reaction
mixture was filtered. The filter cake was dissolved in ethyl
acetate, and washed with aqueous sodium hydroxide solution (10%),
water, sodium thiosulfate (5%), saturated brine. The organic phase
was dried over anhydrous magnesium sulfate to obtain the brown oil
(34.4 g). The crude product was purified by silica gel column
chromatography to obtain the title compound
1-iodo-2-methyl-3-nitro-benzene 74b (30.1 g, yield 81.7%) as a
white solid.
Step 2
3-(2-Iodo-6-nitro-phenyl)-2-oxo-propionic acid
[0544] Sodium ethoxide solution (35 ml, 44 mmol) in an ice-water
bath was added dropwise with a solution of
1-iodo-2-methyl-3-nitro-benzene 74b in ethanol (35 ml, 40 mmol)
under an argon atmosphere. Upon completion of the addition, the
reaction mixture was stirred until lots of brown precipitates were
formed, and added with diethyl oxalate (6 ml, 44 mmol) in one
portion. The reaction mixture was refluxed at 100.degree. C. in an
oil bath for 0.5 hour, added with water (70 ml) and refluxed for
another 1 hour. The reaction mixture was concentrated under reduced
pressure to evaporate ethanol, washed with ethyl acetate (50 ml)
under base condition, adjusted to pH 3 with hydrochloric acid (1M),
and extracted with ethyl acetate (30 ml.times.3). The combined
organic extracts were washed with saturated brine (30 ml), dried
over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure to obtain the title compound
3-(2-iodo-6-nitro-phenyl)-2-oxo-propionic acid 74c (2.94 g) as a
brown oil to be used directly in the next step.
[0545] MS: 334.2[M-1].
Step 3
(2-Iodo-6-nitro-phenyl)-acetic acid
[0546] 3-(2-Iodo-6-nitro-phenyl)-2-oxo-propionic acid 74c (2.086 g,
6.2 mmol) was dissolved in 6 ml of methanol under stirring, and
added with 20 ml of water and 7 ml of hydrogen peroxide to the
solution at room temperature. Upon completion of the addition, the
reaction mixture was stirred at room temperature for 1 hour. After
thin lay chromatography showed the disappearance of starting
materials, the reaction mixture was filtered, extracted with ethyl
acetate (20 ml.times.2). The combined organic extracts were
concentrated under reduced pressure, combined with the above filter
cake, and dried to obtain the title compound
(2-iodo-6-nitro-phenyl)-acetic acid 74d (1.77 g, yield 70%) as a
brown solid.
Step 4
1-Hydroxy-4-iodo-1,3-dihydro-indol-2-one
[0547] (2-Iodo-6-nitro-phenyl)-acetic acid 74d (0.91 g, 3 mmol) was
dissolved in ethanol (30 ml, 95%), and added with palladium on
activated carbon (30 mg, 3%) to the solution. Upon completion of
the addition, the reaction mixture was stirred for 2 hours under a
hydrogen atmosphere. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
filtered, and concentrated under reduced pressure. The residue was
purified by silica gel column chromatography to obtain the title
compound 1-hydroxy-4-iodo-1,3-dihydro-indol-2-one 74e (516 mg,
yield 63.4%) as a white solid.
[0548] MS: 274.1[M-1]
Step 5
5-Fluoro-4-iodo-1,3-dihydro-indol-2-one
[0549] 1-Hydroxy-4-iodo-1,3-dihydro-indol-2-one 74e (326 mg, 1.19
mmol) was dissolved in 24 ml of dichloromethane in a dry
ice-acetone bath, and added slowly with (diethylamino)sulfur
trifluoride (0.16 ml, 1.19 mmol) while maintaining the temperature
at -25.degree. C. Upon completion of the addition, the reaction
mixture was stirred at -25.degree. C. for 15 minutes until it
became clear. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
quenched with saturated sodium bicarbonate solution, added with
saturated brine (30 ml), and extracted with dichloromethane (30
ml.times.3). The combined organic extracts were washed with
saturated brine (30 ml), and dried over anhydrous sodium sulfate to
obtain a yellow solid (284 mg). The crude product was purified by
silica gel column chromatography to obtain the title compound
5-fluoro-4-iodo-1,3-dihydro-indol-2-one 74f (114 mg, yield 34.7%)
as a white solid.
[0550] MS: 276.6[M-1]
Step 6
4-(2,3-Difluoro-phenyl)-5-fluoro-1,3-dihydro-indol-2-one
[0551] 5-Fluoro-4-iodo-1,3-dihydro-indol-2-one 74f (277 mg, 1 mmol)
was dissolved in 10 ml of N,N-dimethylformamide, and added with
2,3-fluoro phenylboric acid (158 mg, 1 mmol), sodium bicarbonate
(168 mg, 2 mmol) and water (10 ml) to the solution under an argon
atmosphere. Upon completion of the addition, the mixture was
stirred to mix well, added with
tetrakis(triphenylphosphine)palladium (109 mg, 0.15 mmol) and
heated to reflux overnight. After thin lay chromatography showed
the disappearance of starting materials, the reaction mixture was
concentrated under reduced pressure, added with hydrochloride acid
solution (10 ml, 1 M), and extracted with ethyl acetate (10
ml.times.3). The combined organic extracts were dried over
anhydrous magnesium sulfate, filtered and concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to obtain the title compound
4-(2,3-difluoro-phenyl)-5-fluoro-1,3-dihydro-indol-2-one 74 g (42
mg, yield 16%) as a gray solid.
[0552] MS: 262.0 [M+1].
Step 7
(Z)-5-(2-(diethylamino)ethyl)-2-(4-(2,3-difluorophenyl)-5-fluoro-2-oxoindo-
lin-3-ylidene)methyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H-
)-one
[0553]
5-(2-Diethylamino-ethyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexahydro-pyrro-
lo[3,2-c]azepine-2-carbaldehyde 1j (73 mg, 0.25 mmol) and
4-(2,3-difluoro-phenyl)-5-fluoro-1,3-dihydro-indol-2-one 74 g (60
mg, 0.23 mmol) were dissolved in 2 ml of ethanol, and added with 35
.mu.l of piperidine to the solution at room temperature. Upon
completion of the addition, the mixture was heated to reflux for 2
hours. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was naturally cooled down
to room temperature, and filtered. The filter cake was washed with
anhydrous ethanol (3 ml.times.2) and dried to obtain the title
compound (Z)-5-(2-(diethylamino)ethyl)-2-((4-(2,3-di
fluorophenyl)-5-fluoro-2-oxoindolin-3-ylidene)methyl)-3-methyl-5,6,7,8-te-
trahydropyrrolo[3,2-c]azepin-4(1H)-one 74 (23 mg, yield 19%) as an
orange solid.
[0554] MS m/z (ESI): 537.2 [M+1]
[0555] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.58 (s, 1H,
pyrrole-NH), 11.178 (s, 1H, indole-NH), 8.24 (s, 1H, --HCO),
7.70-6.998 (m, 5H, --ArH), 6.65 (s, 1H, --CH.dbd.C), 4.36 (m, 2H,
--NCH.sub.2), 2.91 (m, 2H, --NCH.sub.2), 2.54 (s, 3H,
pyrrole-CH.sub.3), 1.2-3.4 (m, 14H, aliphatic H), 2.20 (m, 2H,
--NCH.sub.2).
Example 75
2-((Z)-(4-(2,3-difluorophenyl)-5-fluoro-2-oxoindolin-3-ylidene)methyl)-5-(-
(R)-2-hydroxy-3-morpholinopropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c-
]azepin-4(1H)-one
##STR00193##
[0557]
5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-he-
xahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 53f (84 mg, 0.25 mmol)
and 4-(2,3-difluoro-phenyl)-5-fluoro-1,3-dihydro-indol-2-one 74 g
(60 mg, 0.23 mmol) were dissolved in 2 ml of ethanol, and added
with 35 .mu.l of piperidine to the solution at room temperature.
Upon completion of the addition, the mixture was heated to reflux
for 2 hours. After thin lay chromatography showed the disappearance
of starting materials, the reaction mixture was naturally cooled
down to room temperature, and filtered. The filter cake was washed
with anhydrous ethanol (3 ml.times.2) and dried to obtain the title
compound
2-((Z)-(4-(2,3-difluorophenyl)-5-fluoro-2-oxoindolin-3-ylidene)methyl)-5--
(R)-2-hydroxy-3-morpholinopropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c-
]azepin-4(1H)-one 75 (100 mg, yield 78%) as an orange solid.
[0558] MS ink (ESI): 581.3[M+1]
[0559] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.57 (s, 1H,
pyrrole-NH), 11.12 (s, 1H, indole-NH), 7.704.about.6.98 (m, 5H,
--ArH), 6.65 (s, 1H, --CH.dbd.C), 4.69 (m, 1H, --CHOH), 3.86 (m,
2H, --NCH.sub.2), 2.89 (m, 2H, --NCH.sub.2), 2.33 (m, 2H,
--NCH.sub.2), 3.86.about.2.28 (m, 12H, aliphatic H)
Example 76
(R,Z)-5-(2-hydroxy-3-morpholinopropyl)-3-methyl-2-((4-methyl-2-oxoindolin--
3-ylidene)methyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
##STR00194##
[0561]
5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-he-
xahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 53f (30 mg, 0.09 mmol)
and 4-methyl-1,3-dihydro-indol-2-one (12 mg, 0.08 mmol) were
dissolved in 156 .mu.l of ethanol, and added with 4.4 .mu.l of
piperidine to the solution at room temperature. Upon completion of
the addition, the reaction mixture was stirred at 45.degree. C. for
16 hours. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was naturally cooled down
to room temperature, and filtered. The filter cake was washed with
anhydrous ethanol (1 ml.times.2) and dried to obtain the title
compound
(R,Z)-5-(2-hydroxy-3-morpholinopropyl)-3-methyl-2-((4-methyl-2-oxoindolin-
-3-ylidene)methyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
76 (12 mg, yield 30%) as a orange solid.
[0562] MS m/z (ESI): 465.2 (M+1)
[0563] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.71 (s, 1H,
pyrrole-NH),10.92 (s, 1H, indole-NH), 7.57 (s, 1H, --CH.dbd.C),
7.07 6.77 (m, 3H, --ArH), 4.72 (d, 1H, --OH),3.90 (m, 1H, --CHOH),
3.78 (dd, 1H, seven membered ring outer amide-NCH.sub.2), 3.58 (t,
4H, morpholin intra 2.times.-CH.sub.2O), 3.40 (m, 2H, seven
membered ring --NCH.sub.2), 3.17 (dd, 1H, seven membered ring outer
amide-NCH.sub.2), 2.94 (t, 2H, pyrrole-CH.sub.2), 2.59 (s, 3H,
benzylmethyl), 2.44 (m, 4H, morpholin intra 2.times.CH2N), 2.39 (s,
3H, pyrrole --CH.sub.3), 2.29 (m, 2H, morpholin outer-NCH.sub.2),
2.08 (m, 2H, seven membered ring CH.sub.2--CH.sub.2--CH.sub.2).
Example 77
(R,Z)-5-(2-hydroxy-3-morpholinopropyl)-2-((6-methoxy-2-oxoindolin-3-yliden-
e)methyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
##STR00195##
[0565]
5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-he-
xahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 53f (30 mg, 0.09 mmol)
and 6-methoxyl-1,3-dihydro-indol-2-one (13 mg, 0.08 mmol) were
dissolved in 1560 of ethanol, and added with 4.40 of piperidine to
the solution at room temperature. Upon completion of the addition,
the reaction mixture was stirred at 45.degree. C. for 16 hours.
After thin lay chromatography showed the disappearance of starting
materials, the reaction mixture was naturally cooled down to room
temperature, and filtered. The filter cake was washed with
anhydrous ethanol (1 ml.times.2) and dried to obtain the title
compound
(R,Z)-5-(2-hydroxy-3-morpholinopropyl)-2-((6-methoxy-2-oxoindolin-3-ylide-
ne)
methyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
77 (22 mg, yield 51%) as a yellow solid.
[0566] MS m/z (ESI): 481.2 (M+1)
[0567] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.47 (s, 1H,
pyrrole-NH),10.94 (s, 1H, indole-NH), 7.68 (d, 1H, --ArH), 7.40 (s,
1H, --CH.dbd.C), 6.60 (d, 1H, --ArH), 6.46 (s, 1H, --ArH), 4.70 (d,
1H, --OH),3.89 (m, 1H, --CHOH), 3.77 (dd, 1H, seven membered ring
outer amide-NCH.sub.2), 3.58 (t, 4H, morpholinintra
2.times.-CH.sub.2O), 3.41 (m, 2H, seven membered ring --NCH.sub.2),
3.17 (dd, 1H, seven membered ring outer amide-NCH.sub.2), 2.93 (t,
2H, pyrrole-CH.sub.2), 2.41 (s, 3H, pyrrole-CH.sub.3), 2.34 (m, 4H,
morpholinintra 2.times.-CH.sub.2N), 2.29 (m, 2H, morpholin
outer-NCH.sub.2), 2.07 (m, 2H, seven membered ring
CH.sub.2--CH.sub.2--CH.sub.2).
Example 78
(S,Z)-2-(5-fluoro-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3-morpholino-
propyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
##STR00196##
##STR00197##
[0568] Step 1
(S)-4-Oxiranylmethyl-morpholine
[0569] Morpholine-78a (4.356 m-1.50 mmol) was dissolved in 2.5 ml
of tert-butyl alcohol at room temperature, the solution was cooled
down to 0.degree. C., and added slowly with
(S)-(+)-2-chloromethyl-oxirane (4.02 ml, 50 mmol) to the solution.
Upon the completion of the addition, the reaction system was
naturally warmed up to room temperature and stirred overnight.
After thin lay chromatography showed the disappearance of starting
materials, the reaction system was cooled down to 10.degree. C. in
an ice-water bath, added with a solution of potassium tert-butoxide
in tetrahydrofuran (30 ml, 1.67 mol/L, 50 mmol), the color of the
solution turned from light yellow to white suspension. Upon the
completion of the addition, the reaction mixture was stirred for
another 30 minutes. After thin lay chromatography showed the
disappearance of starting materials, the reaction was stopped. The
reaction mixture was concentrated under reduced pressure, added
with 20 ml of water, extracted with dichloromethane (100
ml.times.3). The combined organic phase was washed with saturated
brine (100 ml.times.1), dried with anhydrous magnesium sulfate,
filtered to remove the drying agent, the filtrate was concentrated
to obtain the title compound (S)-4-oxiranylmethyl-morpholine 78b
(5.52 g, yield 77.2%) as a yellow solid.
[0570] MS m/z (ESI): 144.4 (M+1)
Step 2
(R)-1-Amino-3-morpholin-4-yl-propan-2-ol
[0571] (S)-4-oxiranylmethyl-morpholine 78b (5.52 g, 38.6 mmol) was
added slowly with 395 ml of aqueous ammonia (25%, 5.8 mol) in an
ice-water bath while maintaining the temperature below 0.degree. C.
Upon the completion of the addition, the reaction mixture was
naturally warmed up to room temperature, and stirred for another 18
hours. After thin lay chromatography showed the disappearance of
starting materials, the reaction was stopped. The reaction solution
was concentrated under reduced pressure to remove the reaction
solvent and to obtain the title compound
(R)-1-Amino-3-morpholin-4-yl-propan-2-ol 78c (6.1 g, yield 99%) as
a light yellow oil.
[0572] MS m/z (ESI): 161.3 (M+1)
Step 3
(S)-5-[3-(2-Hydroxy-3-morpholin-4-yl-propylamino)-propyl]-3-methyl-1H-pyrr-
ole-2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester
[0573]
5-(3-methanesulfonyloxy-propyl)-3-methyl-1H-pyrrole-2,4-dicarboxyli-
c acid 2-tert-butyl ester 4-ethyl ester 1g (1.13 g, 2.9 mmol) was
dissolved in 5.6 ml of dichloromethane under stirring, and added
with (R)-1-amino-3-morpholin-4-yl-propan-2-ol 78c (0.93 g, 5.8
mmol) to the solution at room temperature. Upon the completion of
the addition, the reaction mixture was heated to 45.degree. C. for
14 hours in an oil bath. After thin lay chromatography showed the
disappearance of starting materials, the reaction was stopped. The
reaction solution was added with 15 ml of saturated brine,
extracted with dichloromethane (20 ml.times.3), the combined
organic phase was concentrated under reduced pressure, and the
residue was purified by silica gel column chromatography to obtain
the title compound
(S)-5-[3-(2-hydroxy-3-morpholin-4-yl-propylamino)-propyl]-3-methyl-1H-pyr-
role-2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester 78d
(600 mg, yield 72.5%) as a yellow oil.
[0574] MS m/z (ESI): 454.2 (M+1)
Step 4
(S)-5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahydro-1H-p-
yrrolo[3,2-c]azepin-4-one
[0575]
(S)-5-[3-(2-Hydroxy-3-morpholin-4-yl-propylamino)-propyl]-3-methyl--
1H-pyrrole-2 and 4-dicarboxylic acid 2-tert-butyl ester 4-ethyl
ester 78d (580 mg, 1.28 mmol) were dissolved in 6 ml of toluene
under an argon atmosphere, the reaction mixture was cooled in an
ice-water bath, meanwhile a solution of trimethylaluminum in
toluene (1.9 ml, 2 mol/L, 3.84 mmol) was added to the solution.
Upon the completion of the addition, the ice-water bath was
removed, and the reaction solution was heated to reflux for 24
hours. After thin lay chromatography showed the disappearance of
starting materials, the reaction was stopped. The reaction solution
was concentrated under reduced pressure to remove the reaction
solvent, added with 20 ml hydrochloric acid (6 mol/L) and stirred
for 20 minutes, adjusted to about pH 12 with sodium hydroxide
solution (12 mol/L) in an ice-water bath. The resulting mixture was
extracted with dichloromethane (50 ml.times.2), the combined
organic phase was concentrated under reduced pressure, and the
residue was purified by silica gel column chromatography to obtain
the title compound
(S)-5-(2-hydroxy-3-morpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahydro-1H--
pyrrolo[3,2-c]azepin-4-one 78e (300 mg, yield 57.6%) as a white
solid.
[0576] MS m/z (ESI): 308.2 (M+1)
Step 5
(S)-5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexah-
ydro-pyrrolo[3,2-c]azepine-2-carbaldehyde
[0577] (Chloromethylene)dimethylammonium chloride (130 mg, 0.977
mmol) was dissolved in 3 ml of dichloromethane under stirring under
an argon atmosphere, the solution was cooled down to 0.degree. C.
in ice-water bath.
(S)-5-(2-hydroxy-3-morpholin-4-yl-propyl)-3-methyl-5,6,7,8-tetrahyd-
ro-1H-pyrrolo[3,2-c]azepin-4-one 78e (300 mg, 0.977 mmol) was
dissolved in 2 ml of dichloromethane under stirring, the resulting
solution was added to the above solution while maintaining the
temperature below 0.degree. C. Upon the completion of addition, the
reaction mixture was stirred for 20 minutes at room temperature.
After thin lay chromatography showed the disappearance of starting
materials, the reaction solution was added with sodium hydroxide
solution (12 mol/L) to quench the reaction. The reaction solution
was added with 10 ml of saturated brine, extracted with the mixture
solvent of dichloromethane and methanol (V:V=10:1) (100
ml.times.3), the combined organic phase was washed with saturated
brine (100 ml.times.1), dried over anhydrous magnesium sulfate,
filtered to remove the drying agent, the filtrate was concentrated
under reduced pressure, the residue was purified by silica gel
column chromatography to obtain the title compound
(S)-5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,8-hexa-
hydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 78f (200 mg, yield 61%)
as a white solid.
[0578] MS m/z (ESI): 336.2 (M+1)
Step 6
(S,Z)-2-(5-fluoro-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3-morpholino-
propyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
[0579]
(S)-5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 78f (50 mg, 0.149
mmol) was dissolved in 261 .mu.l of ethanol under stirring, and
added with 5-fluoro-1,3-dihydro-indole-2-one (20.28 mg, 0.134 mmol)
and piperidine (7.3 .mu.l, 0.074 mmol) to the solution at room
temperature. Upon the completion of the addition, the reaction
mixture was stirred for 2 hours in dark at 80.degree. C. in an oil
bath. After thin lay chromatography showed the disappearance of
starting materials, the reaction was stopped, and the oil bath was
removed. The reaction system was cooled down to room temperature,
the reaction solution was filtered and the filter cake was dried to
obtain the title compound
(S,Z)-2-((5-fluoro-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3-morpholi-
nopropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
78(40 mg, yield 57%) as a yellow solid.
[0580] MS m/z (ESI): 469.2 (M+1)
[0581] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.73 (s, 1H,
pyrrole-NH),10.91 (s, 1H, indole-NH), 7.79.about.6.84 (m, 3H,
--ArH), 7.56 (s, 1H, --CH.dbd.C),4.73 (d, 1H, --OH),3.90 (m, 111,
--CHOH), 3.76 (dd, 1H, seven-membered ring outer amide-NCH.sub.2),
3.58 (t, 4H, morpholin 2.times.-CH.sub.2O), 3.42 (m, 2H,
seven-membered ring --NCH.sub.2), 3.15 (dd, 1H, seven-membered ring
outer amide --NCH.sub.2), 2.94 (t, 2H, --CH.sub.2C.dbd.C), 2.457
(s, 3H, pyrrole-CH.sub.3), 2.413 (m, 4H, morpholin intra
2.times.-CH.sub.2N), 2.306 (m, 2H, morpholin outer-NCH.sub.2), 2.08
(m, 2H, seven-membered ring CH.sub.2--CH.sub.2--CH.sub.2)
Example 79
(S,Z)-2-((5-chloro-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3-morpholin-
opropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
##STR00198##
[0583]
(S)-5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 78f (40 mg, 0.12
mmol 1) and 5-chloro-1,3-dihydro-indol-2-one (20 mg, 0.12 mmol)
were dissolved in 1.5 ml of ethanol, and added with 6 .mu.l of
piperidine to the solution at room temperature. Upon completion of
the addition, the reaction mixture was stirred at 45.degree. C. for
16 hours. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was naturally cooled down
to room temperature, and filtered. The filter cake was washed with
anhydrous ethanol (1 ml.times.2) and dried to obtain the title
compound
(S,Z)-2-((5-chloro-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3-morpholi-
nopropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
79 (46 mg, yield 79%) as a yellow solid
[0584] MS m/z (ESI): 485.2 (M+1)
[0585] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.679 (s, 1H,
pyrrole-NH),11.008 (s, 1H, indole-NH), 7.994 (s, 1H, --ArH), 7.803
(s, 1H, --CH.dbd.C), 7.159.about.6.869 (m, 2H, --ArH), 4.727 (d,
1H, --OH), 3.90 (m, 1H, --CHOH), 3.76 (dd, 1H, seven-membered ring
outer amide --NCH.sub.2), 3.58 (t, 4H,
morpholin2.times.-CH.sub.2O), 3.418 (m, 2H, seven-membered ring
--NCH.sub.2), 3.15 (m, 1H, seven-membered ring outer amide
--NCH.sub.2), 2.937 (t, 2H, --CH.sub.2C.dbd.C), 2.464 (s, 3H,
pyrrole-CH.sub.3), 2.428 (m, 4H, morpholin intra
2.times.-CH.sub.2N), 2.299 (m, 2H, morpholin outer --NCH.sub.2),
2.076 (m, 2H, seven-membered ring CH.sub.2--CH.sub.2--CH.sub.2)
Example 80
(S,Z)-2-((5-bromo-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3-morpholino-
propyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
##STR00199##
[0587]
(S)-5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 78f (40 mg, 0.12
mmol) and 5-bromo-1,3-dihydro-indol-2-one (25 mg, 0.12 mmol) were
dissolved in 1.5 ml of ethanol, and added with 6 .mu.l of
piperidine to the solution at room temperature. Upon completion of
the addition, the reaction mixture was stirred at 45.degree. C. for
16 hours. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was naturally cooled down
to room temperature, and filtered. The filter cake was washed with
anhydrous ethanol (1 ml.times.2) and dried to obtain the title
compound
(S,Z)-2-((5-bromo-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3-morpholin-
opropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
80 (51 mg, yield 81%) as a yellow solid.
[0588] MS m/z (ESI): 529.1 (M+1)
[0589] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.673 (s, 1H,
pyrrole-NH), 11.014 (s, 1H, indole-NH), 8.120.about.8.115 (s, 1H,
--ArH), 7.807 (s, 1H, --CH.dbd.C), 7.287.about.7.262 (dd, 1H,
--ArH), 6.847.about.6.826 (d, 1H, --ArH), 4.734.about.4.722 (d, 1H,
--OH), 3.90 (m, 1H, --CHOH), 3.792.about.3.748 (dd, 1H, seven
membered ring outer amide-NCH.sub.2), 3.58 (t, 4H,
morpholin2.times.-CH.sub.2O), 3.437.about.3.398 (m, 2H, seven
membered ring --NCH.sub.2), 3.193.about.3.140 (m, 1H, seven
membered ring outer amide-NCH.sub.2), 2.936 (t, 2H,
--CH.sub.2C.dbd.C), 2.465 (s, 3H, pyrrole-CH.sub.3),
2.431.about.2.420 (m, 4H, morpholinintra 2.times.-CH.sub.2N), 2.315
(m, 2H, morpholin outer-NCH.sub.2), 2.09 (m, 2H, seven membered
ring CH.sub.2--CH.sub.2--CH.sub.2)
Example 81
(S,Z)-2-((4-bromo-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3-morpholino-
propyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
##STR00200##
[0591]
(S)-5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 78f (40 mg, 0.12
mmol) and 4-bromo-1,3-dihydro-indol-2-one (25 mg, 0.12 mmol) were
dissolved in 1.5 ml of ethanol, and added with 6 .mu.l of
piperidine to the solution at room temperature. Upon completion of
the addition, the reaction mixture was stirred at 45.degree. C. for
16 hours. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was naturally cooled down
to room temperature, and filtered. The filter cake was washed with
anhydrous ethanol (1 ml.times.2) and dried to obtain the title
compound
(S,Z)-2-((4-bromo-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3-morpholin-
opropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
81 (34 mg, yield 54%) as a yellow solid.
[0592] MS m/z (ESI): 529.3 (M+1)
[0593] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.647 (s, 1H,
pyrrole-NH),11.185 (s, 1H, indole-NH), 8.588 (s, 1H, --CH.dbd.C),
7.238.about.7.218 (d, 1H, --ArH), 7.095.about.7.055 (t, 1H, --ArH),
6.956.about.6.936 (d, 1H, --ArH), 4.739.about.4.726 (d, 1H, --OH),
3.90 (m, 1H, --CHOH), 3.800.about.3.757 (dd, 1H, seven membered
ring outer amide-NCH.sub.2), 3.593.about.3.570 (t, 4H, morpholin
2.times.-CH.sub.2O), 3.446 (m, 2H, seven membered ring
--NCH.sub.2), 3.192 (dd, 1H, seven membered ring outer
amide-NCH.sub.2), 2.956 (t, 2H, --CH.sub.2C.dbd.C),
2.447.about.2.428 (m, 4H, morpholin intra 2.times.-CH.sub.2N),
2.411 (s, 3H, pyrrole-CH.sub.3), 2.301 (m, 2H, morpholin
outer-NCH.sub.2), 2.08 (m, 2H, seven membered ring
CH.sub.2--CH.sub.2--CH.sub.2)
Example 82
(S,Z)-2-((7-bromo-5-fluoro-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3-m-
orpholinopropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
##STR00201##
[0595]
(S)-5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 78f (40 mg, 0.12
mmol) and 7-bromo-5-fluoro-1,3-dihydro-indol-2-one 4b (27 mg, 0.12
mmol) were dissolved in 1.5 ml of ethanol, and added with 6 .mu.l
of piperidine to the solution at room temperature. Upon completion
of the addition, the reaction mixture was reacted at 45.degree. C.
for 16 hours. After thin lay chromatography showed the
disappearance of starting materials, the reaction mixture was
naturally cooled down to room temperature, and filtered. The filter
cake was washed with anhydrous ethanol (1 ml.times.2) and dried to
obtain the title compound
(S,Z)-2-((7-bromo-5-fluoro-2-oxoindolin-3-ylidene)methyl)-5-(2-hydroxy-3--
morpholinopropyl)-3-methyl-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-on-
e 82 (48 mg, yield 73.8%) as a yellow solid.
[0596] MS m/z (ESI): 547.5 (M+1)
[0597] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.665 (s, 1H,
pyrrole-NH), 11.188 (s, 1H, indole-NH), 7.876.about.7.848 (dd, 1H,
--ArH), 7.797.about.7.055 (s, 1H, --ArH), 7.270.about.7.241 (dd,
1H, --ArH), 4.74 (m, 1H, --OH), 3.93 (m, 1H, --CHOH), 3.75 (dd, 1H,
seven membered ring outer amide-NCH.sub.2), 3.583 (t, 4H,
morpholin2.times.-CH.sub.2O), 3.480.about.3.415 (m, 2H,
seven-membered ring --NCH.sub.2), 3.15 (dd, 1H, seven membered ring
outer amide-NCH.sub.2), 2.471 (s, 3H, pyrrole-CH.sub.3), 2.430 (m,
4H, morpholin intra 2.times.-CH.sub.2N), 2.310 (t, 2H, morpholin
outer-NCH.sub.2), 2.086 (m, 2H, seven membered ring
CH.sub.2--CH.sub.2--CH.sub.2)
Example 83
(S,Z)-5-(2-hydroxy-3-morpholinopropyl)-3-methyl-2-((4-methyl-2-oxoindolin--
3-ylidene)methyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
##STR00202##
[0599]
(S)-5-(2-Hydroxy-3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5,6,7,-
8-hexahydro-pyrrolo[3,2-c]azepine-2-carbaldehyde 78f (40 mg, 0.12
mmol 1) and 4-methyl-1,3-dihydro-indol-2-one (18 mg, 0.12 mmol)
were dissolved in 1.5 ml of ethanol, and added with 6 .mu.l of
piperidine to the solution at room temperature. Upon completion of
the addition, the reaction mixture was stirred at 45.degree. C. for
16 hours. After thin lay chromatography showed the disappearance of
starting materials, the reaction mixture was naturally cooled down
to room temperature, and filtered. The filter cake was washed with
anhydrous ethanol (1 ml.times.2) and dried to obtain the title
compound
(S,Z)-5-(2-hydroxy-3-morpholinopropyl)-3-methyl-2-((4-methyl-2-oxoindolin-
-3-ylidene)methyl)-5,6,7,8-tetrahydropyrrolo[3,2-c]azepin-4(1H)-one
83 (35 mg, yield 63.6%) as a yellow solid.
[0600] MS m/z (ESI): 465.2 (M+1)
[0601] .sup.1HNMR (400 MHz, DMSO-d6) .delta. 13.715 (s, 1H,
pyrrole-NH),10.934 (s, 1H, indole-NH),7.572 (s, 1H, --CH.dbd.C),
7.057 (t, 1H, --ArH), 6.840.about.6.821 (d, 1H, --ArH),
6.790.about.6.771 (d, 1H, --ArH), 4.737.about.4.725 (d, 1H, --OH),
3.92 (m, 1H, --CHOH), 3.75 (dd, 1H, seven-membered ring intra amide
--NCH.sub.2), 3.58 (t, 4H, morpholin2.times.-CH.sub.2O), 3.441 (m,
2H, seven-membered ring --NCH.sub.2), 3.15 (m, 1H, seven-membered
ring outer amide-NCH.sub.2), 2.939 (t, 2H, --CH.sub.2CH.dbd.C),
2.594 (s, 3H, pyrrole-CH.sub.3), 2.426 (m, 4H, morpholin intra
2.times.-CH.sub.2N), 2.388 (s, 3H, pyrrole-CH.sub.3),
2.309.about.2.293 (m, 2H, morpholin outer-NCH.sub.2), 2.078 (m, 2H,
seven-membered ring CH.sub.2--CH.sub.2--CH.sub.2)
Biological Assays
Example 1
Inhibition of Cell Proliferation Assay
[0602] The following in vitro assay may be used to determine the
level of activity and effect of different compounds of the present
invention on the proliferation inhibition (cell toxic) of
endothelium growth factor receptor (VEGFR) high expressing Homo
sapiens cancer cell-HUVEC cell.
[0603] The cellular assay described here is to test
anti-angiogenesis and proliferation inhibition activity and effect
of the compounds through VEGFR on the cancer cells in vitro. The
effect and activity is represented by the IC.sub.50 value that
kills the cancer cell. The general procedures for the assay is as
follows: The Homo sapiens cells highly expressing VEGFR are chosen
and seeded to 96-well cell culture plate at a suitable
concentration (exp 5000 cells/ml medium). The cells then are
cultured in carbon dioxide (CO.sub.2) incubator till when they
confluence to about 85%. Then, the cell culture medium is replaced
by fresh one with tested compounds added in it at serial
concentrations (general 6 to 7 concentrations). Then the cells are
put back to the incubator and cultured for continuous 72 hours. 72
hours later, the cell exposed to compounds and control cell are
assayed for their proliferation using Sulforhodamine B (SRB)
method. Compounds IC.sub.50 on tested cells are calculated by the
data of inhibition rates of serial concentrations of the tested
compounds.
Material and Methods:
[0604] a. Dimethyl sulfoxide (Sinophma chemical reagent company,
catalog No. T20050806) [0605] b. HUVEC cells (Purchased from
Institute of biochemistry and cell biology) [0606] c. Falcon 100 mm
cell culture plates (Baton Dickison Labware, Baton Dickison and
company, Catalog No. 18677) [0607] d. Corning 96-well culture
cluster (Corning Incorporated, Catalog No. 3599) [0608] e. Fisher
Pipette (Fisher scientific, Catalog No. 03-692-164) [0609] f.
DMEM/F12 cell medium (Gibco, Catalog No. 12400-024) [0610] g. Fetal
bovine serum, Australia origin (Gibco, Catalog No. 10099-141)
[0611] h. Phosphate Buffered Saline (Gibco, Catalog No. 10010-072)
[0612] i. 0.25% Trypsin-EDTA (Gibco, Catalog No. 25200-056) [0613]
j. Sulforhodamine B (Sigma, Catalog No. 3520-42-1) [0614] k. Acetic
Acid (Sinophma chemical reagent company, Catalog No. T20060508)
[0615] l. Trichloroacetic Acid (Sinophma chemical reagent company,
Catalog No. T20060305) [0616] m. Tris base (Amresco, Catalog No.
0826) [0617] n. Class II A/B3 Biological safety cabinet
(ThermoForma, Catalog No. HB0053-03) [0618] o. Series II water
jacketed CO.sub.2 incubator (ThermoForma, Model: 3111) [0619] p.
Centrifuge (Fisher Scientific Marathon 8 k, Catalog No. 0027-02)
[0620] q. Novastar Plate reader (BMG Labtech, Catalog No. 700-0081)
[0621] r. Orbital Shaker (Qilinbeier, Catalog No. TS-1) [0622]
s.
Protocol:
[0623] The following protocol is used to assay the cell toxic
activity of IC.sub.50 value of tested compounds of the invention on
HUVEC cell: [0624] 1. HUVEC cells were grown in growth media
(DMEM/F12, supplemented with 10% FBS) in 100 mm corning culture
plates till confluence at 37.degree. C., 5% CO.sub.2. [0625] 2.
HUVEC cells were washed in 100 mm plates with FBS, then the cells
were harvested by trypsinization and seeded to corning 96-well cell
culture plates at concentration of 50000 cells/ml, leaving 6
wells/each plate empty as background. [0626] 3. The cells were
grown in 96-well plates at 37.degree. C., 5% CO.sub.2, till 85%
confluence. [0627] 4. The compounds stock solution was prepared,
using DMSO to solve candidate compounds to a concentration of 20
mM. Then DMSO was used to dilute the stock solution to a serious
concentration of tested compounds solution (namely, 2 mM, 1 mM, 0.2
mM, 20 .mu.M, 2 .mu.M, 0.2 .mu.M). [0628] 5. Cell culture medium
was used (in this case, DMEM/F 12, supplemented with 10% FBS) to
dilute the compounds solution prepared above. Each DMSO serial
concentration compound solution was diluted by 20 times with
culture medium by adding 5 .mu.l DMSO compound solution to 95 .mu.l
culture medium, then mixed well by vortex. This promised that the
DMSO concentration at that the HUVEC cell will exposed to will not
surpass 0.5%. [0629] 6. After HUVEC cell had attached to the dish
bottom and confluence about 85%, the culture medium was replaced by
fresh one with fresh DMEM/F12, supplemented with 10% FBS. Each well
was added 180 .mu.l medium, then 20 .mu.l medium solution of tested
compounds prepared at step 5 was added to each well. For negative
control group cell, 20 .mu.l culture medium containing 0.5% pure
DMSO was added. So, HUVEC cells were exposed to each tested
compound at a serial final concentration of 100 .mu.M, 10 .mu.M, 5
.mu.M, 1 .mu.M, 0.1 .mu.M, 0.01 .mu.M, and 0.001 .mu.M. [0630] 7.
The culture plates were put back to incubator, and cultured for 72
hours at 37.degree. C., 5% CO.sub.2. [0631] 8. 72 hours later,
cultures were removed from incubator into sterile work area. [0632]
9. The fixative (50% Trichloroacetic Acid-TCA) was prepared by
adding reagent grade water to the TCA, fixing the cells by gently
layering 50 .mu.l of cold TCA solution on top of the growth medium.
[0633] 10. The plates was incubated for 1 hour at 4.degree. C. and
then rinsed with water several times to remove TCA, serum proteins,
etc. Plates were air dried and stored until use. Bland background
optical density was measured in wells incubated with growth medium
without cells. [0634] 11. 0.4% Sulforhodamine B solution was
prepared by using 10% acetic acid solution. 50 .mu.l sulforhodamine
B solution was added to each well of 96-well plates. [0635] 12. The
cells were allowed to stain for 30 minutes. [0636] 13. The wash
solution of 10% acetic acid was prepared. At the end of the
staining period, the stain was removed and the cells were rinsed
quickly with 1% acetic acid. Repeat until unincorporated dye was
removed. Wash times were kept to a minimum to reduce desorption of
protein-bound dye. After being rinsed, the cultures were air dried.
[0637] 14. The incorporated dye was then dissolved in a volume of
Sulforhodamine B. Solubilization solution (10 mM Tris) was equal to
the original volume of culture medium. Then cultures were allowed
to stand for 5 minutes at room temperature, and the mixing of the
dye was enhanced by gentle stirring in a gyratory shaker. [0638]
15. The absorbance was measured by spectrophotometry at a
wavelength of 565 nm. The background absorbance of 96-well plates
at 690 nm was measured and subtracted from the measurement at 565
nm. [0639] 16. The inhibition rate (IR) was calculated as
follows:
[0639] IR=100.times.(Absorbance of control cells-Absorbance of
cells exposed to tested compounds at each concentration)/Absorbance
of control cells %. [0640] The IC.sub.50 value can be derived from
the IRs of compounds at different concentration gradients.
The Activity of the Compounds of the Invention:
[0641] The biological activity of the compounds of the invention is
tested using the assay described above. The IC.sub.50 values are
measured and showed in table 1 below:
TABLE-US-00002 Example No. IC.sub.50 (VEGFR/HUVEC) (.mu.M) 1 0.09 2
0.288 3 0.1 4 0.404 5 0.1 6 0.28 8 0.308 9 0.068 10 0.123 11 0.207
12 0.3 13 0.804 14 0.457 15 1.2 16 0.21 17 0.129 19 0.146
Example 2
VEGFR-2 Kinase Assay
[0642] This assay is used to measure the in vitro kinase activity
of recombinant human VEGF-R2 in an ELISA assay.
Materials and Reagents:
[0643] a. Wash Buffer (PBS-T Buffer): 1.times.PBS (137 mM NaCl, 2.7
mM KCl, 4.3 mM Na.sub.2HPO.sub.4, 1.4 mM KH.sub.2PO.sub.4, the pH
adjusted to 7.2) and 0.05% Tween-20. [0644] b. 1% Bovine Serum
Albumin (BSA, Calbiochem #136593) in PBS-T Buffer. [0645] c. Stop
Buffer: 50 mM EDTA, pH 8.0. [0646] d. DELFIA.RTM. Europium-labeled
Anti-mouse IgG (PerkinElmer Life Sciences #ADO 124). [0647] e.
DELFIA.RTM. Enhancement Solution (PerkinElmer Life Sciences
#1244-105). [0648] f. DELFIA.RTM. Streptavidin coated, 96-well,
yellow plate (PerkinElmer Life Sciences #AAAND-0005). [0649] g.
recombinant human VEGF-R2 kinase (supplied in 50 mM Tris-HCl (pH
8.0), 100 mM NaCl, 5 mM DTT, 15 mM reduced glutathione and 20%
glycerol) (Cell signaling technology #7787). [0650] h. 10 mM ATP
solution (Cell signaling technology #9804). [0651] i.
Biotin-Gastrin Precursor (Tyr87) Peptide (Cell signaling technology
#1310). [0652] j. Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Cell
signaling technology #9411). [0653] k. HTScan.TM. Tyrosine Kinase
Buffer (4.times.) [0654] 1.times. Kinase Buffer: [0655] 60 mM HEPES
[0656] 5 mM MgCl.sub.2 [0657] 5 mM MnCl.sub.2 [0658] 3 .mu.M
Na.sub.3VO.sub.4 [0659] (Cell signaling technology #9805). [0660]
1. 1.25 M DTT (1000.times.) (Cell signaling technology).
Procedure:
[0661] The following protocol was used: [0662] 1. The test compound
was diluted with DMSO to the desired final assay concentration. 1
.mu.l of test compound, the negative control (sample which does not
receive any test compound), 1 .mu.l DMSO were added for each assay.
[0663] 2. 6 .mu.M substrate peptide (Tyr87) was diluted with
dH.sub.2O (1:1), 15 .mu.l was added to every assay. [0664] 3.
VEGFR-2 enzyme was immediately transferred from -80.degree. C. to
ice, and the enzyme was allowed to thaw on ice. [0665] 4. 2.2 .mu.g
VEGFR-2 enzyme was taken to the enzyme tube. [0666] 5. 10 .mu.l of
DTT (1.25 M) was added to 2.5 ml of 4.times. HTScan.TM. Tyrosine
Kinase Buffer (240 mM HEPES pH 7.5, 20 mM MgCl.sub.2, 20 mM
MnCl.sub.2, 12 .mu.M Na.sub.3VO.sub.4) to prepare DTT/Kinase
buffer. [0667] 6. 0.75 ml of DTT/Kinase buffer was transferred to
each enzyme tube to prepare 4.times. reaction cocktail, and 7.5
.mu.l 4.times. reaction cocktail was added to every assay. [0668]
7. 2 .mu.l ATP (10 mM) was added to 498 .mu.A dH.sub.2O, 7.5 .mu.l
was added for every assay. Final Assay Conditions for a 30 .mu.l
Reaction [0669] 60 mM HEPES pH 7.5 [0670] 5 mM MgCl.sub.2 [0671] 5
mM MnCl.sub.2 [0672] 3 .mu.M Na.sub.3VO.sub.4 [0673] 1.25 mM DTT
[0674] 10 .mu.M ATP [0675] 1.5 .mu.M substrate peptide [0676] 22 ng
VEGFR-2 Kinase [0677] 8. The reaction tube was incubated at
25.degree. C. for 30 minutes. [0678] 9. 30 .mu.l/assay stop buffer
(50 mM EDTA, pH 8.0) was added to stop the reaction. [0679] 10. 25
.mu.l of each reaction and 75 .mu.l dH.sub.2O/well was transferred
to a 96-well streptavidin coated plate, with shaking at room
temperature for 60 minutes. [0680] 11. Each well was washed three
times with 200 .mu.l PBS-T buffer. Plate was patted on paper towel
to remove excess liquid. [0681] 12. Primary antibody,
Phospho-Tyrosine mAb (P-Tyr-100) was diluted, 1:1000 in PBS-T
buffer with 1% BSA, and 100 .mu.l diluted primary antibody was
added to each well. [0682] 13. The reaction tube was incubated,
with shaking at room temperature for 60 minutes. [0683] 14. Washing
was carried out as described above in step 11. [0684] 15. Europium
labeled anti-mouse IgG was diluted 1:500 in PBS-T buffer with 1%
BSA, 100 .mu.l diluted antibody was added to each well. [0685] 16.
The reaction tube was incubated, with shaking at room temperature
for 30 minutes. [0686] 17. Each well was washed five times with 200
.mu.l PBS-T buffer. Plate was patted on paper towel to remove
excess liquid. [0687] 18. 100 .mu.l/well DELFIA.RTM. Enhancement
Solution was added. [0688] 19. The reaction tube was incubated,
with shaking at room temperature for 5 minutes. [0689] 20. The
fluorescence emission was detected at 615 nm with appropriate
Time-Resolved Plate Reader. [0690] Calculate the inhibition rate:
IR (%)=100-100*(X-B)/(N-B) [0691] X=the fluorescence value of the
well contained test compound [0692] N=negative control [0693]
B=blank
[0694] The IC.sub.50 value can be derived from the IRs of compounds
at different concentration gradients.
The Activity of the Compounds of the Invention
[0695] The biochemical activity of the compounds of the invention
is tested using the assay described above. The IC.sub.50 values are
measured and showed in table 2 below:
TABLE-US-00003 Example No. IC.sub.50 (VEGFR-2/bio) (.mu.M) 1 0.014
2 0.021 3 0.28 4 0.041 5 0.0085 6 0.023 7 0.69 8 0.115 9 0.49 10
0.014 11 0.001 12 0.001 13 0.003 14 0.004 15 0.002 16 0.0013 17
0.001 18 0.01 19 0.012 20 0.072 21 0.16
Example 3
Inhibition of Receptor Tyrosine Kinases Activity Assay
[0696] The in vitro kinase activity of Receptor tyrosine including
kinases of VEGFR-2, c-Kit, PDGFR.beta., Flt1, VEGFR-3, Ret, or
Flt3, was tested by the following assay.
[0697] Kinases used in this assay are human-derived recombinant
proteins. Their article number information is given as follows:
TABLE-US-00004 VEGFR-2: invitrogen PV3660 c-Kit: invitrogen PV3081
PDGFR.beta.: invitrogen PV3082 Flt1: invitrogen PV3666 VEGFR-3:
invitrogen PV4129 Ret: invitrogen PV3819 Flt3: invitrogen
PV3182
[0698] The following assay may be used to determine the activity of
the compound of Example 53 for inhibiting kinase activity of
VEGFR-2, c-Kit, PDGFR.beta., Flt1, VEGFR-3, Ret, or Flt3. The half
maximal inhibitory concentration IC.sub.50 (the concentration of
the tested compound showing 50% inhibition of the enzyme activity)
of each kinase was determined by incubating several different
concentrations of the tested compound with a specific enzyme and
substrate. Each kinase was reacted with peptide substrate and
tested compound in a buffer solution containing 60 mM HEPES
(pH7.5), 5 mM MgCl.sub.2, 5 mM MnCl.sub.2, 3 .mu.M
Na.sub.3VO.sub.4, 1.25 M DTT (1000.times.) and 20 .mu.M ATP at
25.degree. C., for 45 minutes. The kinase activity was determined
by using a Time-Resolved fluorescence method.
The Activity of the Compounds of the Invention:
[0699] The biological activity of the compound of Example 53 was
tested by using the assay described above. The IC.sub.50 values
were measured and showed in table 3 below.
TABLE-US-00005 TABLE 3 Inhibition activity of the compound of
Example 53 on Receptor tyrosine kinases Example 53 Kinase VEGFR-2
c-Kit PDGFR.beta. Flt1 VEGFR-3 Ret Flt3 IC50 0.5 .+-. 0.4 3.3 .+-.
3.6 41.5 .+-. 22.7 30.9 .+-. 5.7 9.6 .+-. 7.8 182.5 .+-. 6.4 19.3
.+-. 3.1 nM, mean .+-. SD
[0700] Conclusion: the compound of Example 53 had obvious activity
for inhibiting the kinase activity of receptor tyrosine including
kinases of VEGFR-2, c-Kit, PDGFR.beta., Flt1, VEGFR-3, Ret, or
Flt3.
Example 4
Inhibition of Cancer Cell Proliferation Assay
[0701] The cellular assay described here is to test the
proliferation inhibition activity of the compound of Example 53 on
the cancer cells in vitro using Sulforhodamine B (SRB) method. The
cancer cells include HUVEC (human umbilical vein endothelial
cells), HT-29 (human colon cancer cells), Mo7e (human
megakaryoblastic leukemia cell line), A431 (human epidermoid
carcinoma cells), NCI--H526 (small-cell lung cancer cells), and
SK--BR-3 (human breast cancer cells).
[0702] Article number information of cells used in this assay is
given as follows:
[0703] HUVEC cancer cell: American Type Culture Collection, Cat.
No. CRL-1730
[0704] HT-29 cancer cell: Cell Bank, Chinese Academy of Sciences,
TCHu103;
[0705] Mo7e cancer cell: originally described by Avanzi et al.
((1988) Br. J. Haematol. 69:359-366), was obtained from Genetics
Institute (Boston, Mass.) and was maintained in Iscove's modified
Dulbecco's medium (Life Technologies, Inc.);
[0706] A431 cancer cell: American Type Culture Collection, Cat. No.
CRL-2592;
[0707] NCI--H526 cancer cell: American Type Culture Collection,
Cat. No. CRL-5811;
[0708] SK--BR-3 cancer cell: Cell Bank, Chinese Academy of
Sciences, TCHu225.
[0709] The following in vitro assay is to determine the activity of
the tested compound for inhibiting the proliferation of the cancer
cells mentioned above. The activity is represented by the IC.sub.50
value. The general procedures of the assay are given as follows:
Cancer cells mentioned above were chosen and seeded to 96-well cell
culture plate at a suitable concentration (e.g., 5000 cells/mL
medium). These cells then were incubated in carbon dioxide
(CO.sub.2) incubator until they reached 85% confluency. Then, the
cell culture medium was replaced by fresh one with tested compound
added in it at serial concentrations (general 6 to 7
concentrations). Then the cells were put back to the incubator and
cultured continuously. 72 hours later, the activity of the tested
compound for inhibiting the cell proliferation was determined by
using Sulforhodamine B (SRB) method. IC.sub.50 value on tested
cells is calculated by the data of inhibition rates of serial
concentrations of the tested compound.
The Activity of the Compounds of the Invention:
[0710] The biological activity of the compounds of the invention
was tested by using the assay described above. The IC.sub.50 values
were measured and showed in table 4 below.
TABLE-US-00006 TABLE 4 Proliferation inhibition activity of the
compound of Example 53 on the cancer cells in vitro Example IC50
(.mu.M, mean .+-. SD) No. HUVEC HT-29 Mo7e A431 NCI-H526 SK-BR-3 53
1.1 2.4 .+-. 1.6 0.8 .+-. 0.2 2.0 .+-. 0.5 8.3 .+-. 0.3 7.8 .+-.
1.5
[0711] The results showed that the compound of Example 53 obviously
inhibited the proliferation of HUVEC, Mole highly expressing
receptor tyrosine kinase VEGFR, c-Kit, IC.sub.50 values are
respectively 1.1 .mu.M and 0.8 .mu.M. In addition, the compound of
Example 53 inhibited the proliferation of A431, SK--BR-3 highly
expressing EGFR, HER2, and inhibited the proliferation of NCI--H526
highly expressing c-kit.
[0712] In summary, the assay proveed the method of the present
invention can inhibit protein kinase VEGFR, c-Kit, EGFR, HER2.
Moreover, the method of the present invention obviously inhibited
the proliferation of colon cancer cells, leukemia cells, squamous
cell carcinoma, small cell lung cancer cells, breast cancer
cells.
Example 5
Anticancer Pharmacodynamic Assay in vivo
[0713] The test method and steps are the same with those disclosed
in "PHARMACODYNAMIC ASSAYS IN VIVO" of the present invention. The
therapeutic effects of the compound of Example 53 against
xenografts of colon cancer, non-small cell lung cancer, liver
cancer, melanoma, renal cancer in nude mice are estimated.
[0714] Article number information of cells used in this assay is as
follow:
[0715] HT-29(colon cancer): Cell Bank, Chinese Academy of Sciences,
TCHu103;
[0716] Calu-3(non-small cell lung cancer): Cell Bank, Chinese
Academy of Sciences, TCHu157;
[0717] NCI--H460 (non-small cell lung cancer): Cell Bank, Chinese
Academy of Sciences, TCHu205;
[0718] Bel-7402(liver cancer): Cell Bank, Chinese Academy of
Sciences, TCHu10;
[0719] A375 (melanoma): Cell Bank, Chinese Academy of Sciences,
TCHu4;
[0720] Caki-1 (renal cancer): Cell Bank, Chinese Academy of
Sciences, TCHu135.
[0721] The test results were shown in table 5.
TABLE-US-00007 TABLE 5 Anticancer activity of the compound of
Example 53 in vivo Original volume of Cancer Dose Admini- tumor T/C
cells Drug (mg/kg) stration (mm.sup.3) (%) Note HT-29 Control
solvent p.o, d0-20 146 .+-. 8 Example 53 15 p.o, d0-20 147 .+-. 7
49.4 Example 53 30 p.o, d0-20 151 .+-. 4 48.5 Example 53 60 p.o,
d0-20 156 .+-. 18 31.6 Calu-3 Control solvent p.o, d0-20 227 .+-.
27 Example 53 15 p.o, d0-20 223 .+-. 29 26.3 Example 53 30 p.o,
d0-20 252 .+-. 44 14.7 3/6(PR) Example 53 60 p.o, d0-20 200 .+-. 37
16.1 3/6(PR) NCI- Control solvent p.o, d0-13 184 .+-. 37 H460
Example 53 15 p.o, d0-13 185 .+-. 27 41.1* Example 53 30 p.o, d0-13
181 .+-. 21 46.3* Example 53 60 p.o, d0-13 176 .+-. 28 58.4* Bel-
Control solvent p.o d0-17 119 .+-. 20 7402 Example 53 15 p.o d0-17
114 .+-. 15 39.8 Example 53 30 p.o d0-17 120 .+-. 21 29.2 Example
53 60 p.o d0-17 119 .+-. 12 13.3 A375 Control solvent p.o d0-16 160
.+-. 10 Example 53 15 p.o d0-16 147 .+-. 25 82.5 Example 53 30 p.o
d0-16 146 .+-. 7 58.5 Example 53 60 p.o d0-16 155 .+-. 6 46.4
Caki-1 Control solvent p.o, d0-22 242 .+-. 68 Example 53 15 p.o,
d0-22 292 .+-. 78 73.0 Example 53 30 p.o, d0-22 303 .+-. 58 45.5
Example 53 60 p.o, d0-22 212 .+-. 62 36.8 *P < 0.01 vs Control;
Control: n = 12; Treatment group: n = 6; PR: partial regression;
p.o: per os; d0-17: from 0 day to 17 day; T/C (%): relative tumor
volume (%).
[0722] The results showed the anticancer activity of the compound
of Example 53 against xenografts of colon cancer (HT-29), non-small
cell lung cancer (NCI--H460, Calu-3), liver cancer (Bel-7402),
melanoma (A375), renal cancer (Caki-1) in nude mice in vivo. The
compound of Example 53 had obvious anticancer activity against
xenografts listed above, reduced the tumor volume, and it was a
broad spectrum antitumor medicine.
Pharmacodynamic Assays In Vivo
The Therapeutic Effects of the Compound of Example 63 Against
Xenografts of HT-29 Human Colon Cancer in Nude Mice
1. ABSTRACT
[0723] The therapeutic effects of Example 63 against xenografts of
HT-29 human colon cancer in nude mice were estimated. Continuous
oral the compound of Example 63 markedly inhibited the growth of
HT-29 human colon cancer and reduced the tumor volume, mice can be
well tolerant to the compound.
2. PURPOSE
[0724] The therapeutic effects of the compound of Example 63
against xenografts of HT-29 human colon cancer in nude mice were
estimated.
3. TEST DRUG
[0725] Drug name and batch: the compound of Example 63 is a yellow
powder.
[0726] Preparation method: the compound of Example 63 was prepared
to corresponding concentration with distilled water.
4. TEST ANIMALS
[0727] Six to seven-week-old BALB/cA-nude female mice was purchased
from Slaccas Experimental Animal. Certificate No.: SCXK(Shanghai)
2003.about.0003. Raising Conditions: SPF level.
5. TEST PROTOCOL
[0728] Nude mice were hypodermic inoculated HT-29 human colon
cancer cell. After tumors grew to 100-300 mm.sup.3, mice were
randomly divided into teams (d0). The dose and dosage regimen were
shown in table 6. The volume of tumors and weigh of the mice were
measured for 2-3 times per week. The calculation formula of the
volume of tumor (V) is: V=1/2.times.a.times.b.sup.2, a: length of
tumor, b: width of tumor.
TABLE-US-00008 TABLE 6 The therapeutic effect of the compound of
Example 63 against xenografts of HT-29 human colon cancer in nude
mice. Animal TV Dose Numbers (X .+-. SD, mm.sup.3) RTV T/C Group
(mg/kg) Administration d0/dn d0 dn X .+-. SD (%) Control p.o 9/9
291 .+-. 66 729 .+-. 298 2.54 .+-. 1.01 Example 63 40 d0~12 5/5 364
.+-. 46 300 .+-. 154 0.80 .+-. 0.42 31.5* d0: the time of the first
dosage; dn: the 13th day after the first dosage; TV: volume of
tumor; RTV: relative tumor volume; *P < 0.01 vs control.
6. CONCLUSION
[0729] Example 63 obviously inhibited the growth of HT-29 human
colon cancer and reduced the tumor volume; mice can be well
tolerant to the compound without obvious toxicity.
* * * * *