Note: Descriptions are shown in the official language in which they were submitted.
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NEUROPROTECTIVE AND ANTI-PROLIFERATIVE COMPOUNDS
FIELD OF THE INVENTION
This invention features pyrrolo-(3-carboline derivatives and derivatives of 3-
(1H indol-3-yl)-
1H pyrrole-2,5-dione which are useful in prevention and treatment of
degenerative and inflammatory
diseases of the central and peripheral nervous systems, by inhibiting axonal
degradation and/or
neuronal apoptosis, as well as in the treatment and prevention of cancer and
inflammation by inducing
apoptosis in proliferating cells.
BACKGROUND OF THE INVENTION
The regulation of cellular responses to ischemic, excitotoxic, pathogenic or
chemotoxic
stresses in the central and peripheral nervous systems (CNS and PNS,
repectively), including for
example, the brain, the spinal cord, the eye, and the peripheral sensory and
motor neurons, is a major
frontier of modern medicine. Neurons are non-proliferating cells whose
progressive or abrupt loss
can result in diseases exemplified by Alzheimer's disease (AD), Parkinson's
disease (PD),
Huntington's disease (HD), amyotrophic laterial sclerosis ("ALS" or "Lou
Gehrig's disease"), and
stroke. These diseases and disorders are individually or collectively referred
to herein as "central
neurodegenerative diseases." Selected symptoms resulting from these diseases
include memory loss,
loss of cognitive function, loss of gross and fine motor control, and
blindness. Peripheral neuronal
loss or neurite damage results in sensory loss exemplified by pain or
discomfort, sensorimotor defects,
and paralysis.
The incidence of central neurodegenerative diseases increases with age. For
example, less
than 5% of the population under the age of 65 displays signs of AD. An
exponential increase is
observed over the age of 65, with as much as 47% of the population displaying
some form of AD over
the age of 85. Many factors (etiological agents) are responsible for the
initiation of neurodegenerative
conditions, factors as varied as genetic DNA damage or loss in the
mitochondria, abnormal amyloid
processing, oxidative stress following ischemia and reperfusion, and loss of
neurotrophic support for
the nerve cells. The mode of action ultimately underlying such irreversible
neuronal loss involves
programmed cell death, or apoptosis. Preventing neuronal apoptosis and neurite
disfunction represents
a new, broad-spectrum, approach to the treatment of progressive central
neurological disorders
Various other neurodegenerative diseases related to the peripheral nervous
system, herein
referred to as "peripheral neuropathies", are characterized by the loss of
feeling, experiencing pain,
and even paralysis of or in the extremities. These peripheral neuropathies
result from disease states
such as ALS, Multiple Sclerosis, AmS, diabetes, and various neuropathies
induced by
chemotherapeutic treatments such as cisplatin, vinblastine and taxane (Taxol~
and TaxotereTM)
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2
treatment for cancer therapy, and D4T for the treatment of HIV (Human
Insufficiency Virus). In most
of these cases, progressive loss of axonal function occurs initially,
resulting in severe symptoms,
followed by the apoptotic loss of the neuron. In these cases, inhibiting
neuronal apoptosis and or
axonal degradation is a new approach to treating these diseases.
Axecently discovered family of genes, known as the IAPs (Inhibitor of
Apoptosis Proteins),
potently inhibit apoptosis in most mammalian cell lines. Members of the IAP
family, specifically
NAIP, HIAP1,2 and XIAP, are used as survival factors by both neurons and
cancer cells to resist
intrinsic apoptosis.
NAIP's (Neuronal Apoptosis Inhibitory Protein) primary function appears to be
the
10' regulation of neuronal apoptosis (Xu, D.G. et al. Nature Medicine 1997, 3,
997). NAIP is primarily
expressed in neurons where it serves to protect these post-mitotic cells
against environmental and
metabolic stresses that lead to premature apoptosis of the neuron. Indeed,
deletions in the NAIP gene
were found to be causally related to the severity of the childhood genetic
neuromuscular disease,
Spinal Muscular Atrophy (SMA).
Enhancement of NAIP expression in the brain was achieved through the systemic
in vivo
administration of a neuroprotective alkaloid, K252a (for the isolation and
identification of K252a see
Sezaki, M. J. A~atibiot. 1986, 39, 1066, US 6,020,127). In vivo studies
demonstrated increased
expression of NAIP in hippocampal neurons after K252a administration to rats.
These results
correlate well with increased protection to ischemic insults provided by this
compound (see Xu, D.G.
et al. Natm°e Medicifae 1997, 3, 997). Knock-out mice lacking the
expression of NAIP displayed
dramatic neuronal sensitivity to such ischemic insults.
The exact mechanism by which K252a upregulates NAIL' gene expression is not
known.
However, it is lrnown that K252a inhibits several classes of protein kinases.
The X-ray crystal
structure of a closely related natural product alkaloid, staurosporine, when
bound to the protein
lcinases CDK2 and cAPK confirmed that staurosporine acts as a competitive
inhibitor for the
conserved binding site of adenosine triphosphate (ATP), which is found in all
lrnown protein kinases
enzymes (for a review see Toledo and Lydon Structure 1997, 5, 1551). Several
groups have suggested
that K252a and its structural analogues, the indolocarbazoles, also bind to
the ATP binding site of
various protein kinases. A large number of natural products related to the
K252a structure
(indolocarbazoles) also inhibit various serine-threonine protein kinases. Most
of these compounds
have undesirable neuronal cytotoxic effects due to their lack of kinase
specificity. Non-specific
kinase inhibitory compounds can interrupt the neuronal survival signaling
pathways for example, by
inhibiting PKB or PKC. In fact, protein kinase deregulation has been
implicated as a contributing
factor to various neurodegenerative disorders (Bradshaw, D. et al. Agents afZd
Actiofas 1993, 38, 137;
Knusel B. & Hefti F. J. Neu~ochenz. 1992, 59, 1987). This class of compounds
is typified by the
following compounds:
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WO 01/87887 PCT/CA01/00718
3
H H R
i ~i i
N. r,
R~ ~. /-\ ~ ,R
HsCi.,
MeOZC'
NHMe
(+)-K252a; R=H Staurostorine; R = H Rebeccamycin; R = H, X = CI NB 506
CEP 1347; R=CHZSEt UNC 01; R = OH R-3; R = OH, X = H
K252a displays significant neuronal cytotoxicity at moderate doses in vitro
which preclude
the measurement of upregulation of NAIP gene expression as a true indication
of its neuroprotective
mechanism in either cultured neuroblastoma cells or cerebellar granule neurons
(CGN). These
findings suggest that highly specific compounds will be required in order to
have pharmaceutical
potential in regulating pro-apoptotic action in various diseases.
Various cancers and cell lines, including colon, lung, and breast, display
elevated levels of
other IAPs, including HIAP1,2 and XIAP, as either mRNA andlor protein (US
5,919,912). Scientist
at Aegera Therapeutics Tnc. have shown that the down-regulation of the IAPs in
cancer cells can
effectively shift the chemotherapeutic dose response required to kill such
cancer cells, in the case of
HAIP 1, or to kill cancer cells outright, in the case of XIAP (US pat
application. Compounds that
down-regulate the expression of these genes would therefore be useful in
treating cancers. In several
cases, the cytotoxic properties of the indolocarbazoles have been exploited to
affect a therapeutic use
in cancer eg. Staurosporine, UNC O1, Rebeccamycin and NB 506 amongst others.
Specifically, UNC
O1 down regulates XIAP expression in B-cell chronic lymphocytic leukemia cell
lines, inducing
apoptosis (Kitada, S. et al Blood 2000, 96, 393).
Indolocarbazole Alkaloids Staurosporine and K252a.
Various derivatives of the natural products Staurosporine and K252a have been
described for
the treatment of neurodegenerative disorders. US Patent No. 6,013,646 to Roder
et al. (issued
January 1 l, 2000) discloses K252a derivatives incorporating a carbon at the
tetrahydrofuran oxygen
position of the K252a sugar moiety prevents tau hyperphosphorylation by the
direct inhibition of the
ERK family of protein kinases, also known as the MAP kinases. Tau
hyperphosphorylation results in
the destabilization of regular microtubular organization and the formation of
neurofibular tangles
(Iqbal, K. et al. FEBS Lett., 1994, 349, 104; Garver, T. D. et al., J.
Neurosci. Res., 1996, 44, 12).
Neurofibulary tangles are associated with neurodegenerative diseases such as
AD and PD. A report
by Murakata, C. et al. (J. Med Chern. 1997, 40, 1863) established that the
semisynthetic K252a
analogue, CEP 1347, is a selective neurotrophic agent in which the undesirable
TrkA and PKC
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4
inhibitory activities have been reduced, as demonstrated in a ChAt assay.
Additionally, this class of
compounds appears to inhibit the production of TNF-a (tumour necrosis factor),
which is intimately
involved in the initiation of neuronal apoptosis. At the same time CEP 1347
and related compounds
upregulated the production of IL-1 (3 (Mallamo et al, W096/31515; Hudkins et
al, WO 97/46565;
Engber et al, W097/49406). Maroney, A. C, et al. showed that CEP 1347
inhibited JNKl activation
(J. Neur°osci.,1998, 18, 104).
Other indolocarbazole derivatives are disclosed by Gliclcsman, M. A. et al.
(WO 95 07911),
Lewis, M. E. (WO 94 02488), Lewis, M. E. et al. (LTS Patents No. 5,756,494,
No. 5,741,808, and No.
5,621,101). Indolocarbazole derivatives have also been reported for use in
treatment of cancer (EP 0
323 171, EP 0 643 966, US 4,923,986, US 4,877,776, WO 94 27982), as
antimicrobial agents
(Prudhomme et al, J. Antibiotics, 1994, 47, 792) and in the treatment of
hypertension (Hachisu et al.
Life Sciences 1989, 44, 1351).
A variety of synthetic procedures have been reported in the literature for the
preparation of
bis(indolyl)pyrrole-2,5-diones and indolocarbazoles. See, for example, Bit et
al., J. Med. Chern.,
1993, 63, 21; Bergman et al., Tetrahedron Lett., 1987, ~8, 4441; Davis et al.,
Tetrahed~oya Lett., 1990,
31, 2353, 5201; Faul, M. M, et al., Tett°ahedron Lett., 1999, 40, 1109;
Faul M.M, et al. US Patents
No. 5,859,261, No. 5,919,946, and No. 6,037,475. For a general review of the
chemistry and
properties of these alkaloids see Gribble, G. W.; Berthel, S. J. "Studies in
Natural Products
Chemistry", 1993, 12, 365. For synthetic studies see Wood, J. L. et al. J. Am.
Chena. Soc. 1997, 119,
9641; and Danishefsky, S. et al. J. Am. Clzem. Soc. 1996, 118, 2825.
A class of indolocarbazoles having fused imidazolyl ring systems are known as
the
granulatimides. Iso-granulatimide has been shown to be an effective G2 check
point inhibitor in p53
deficient cancer cell lines, suggesting its potential in cancer chemotherapy
(PCT WO99/47522, Sept
23 1999). Some members of this class are illustrated below.
H H
O N O O N O
NH
N
N N N
H H
granulatimide iso-granulatimide
H H
O N O
1l ~ ~ ~ N JN
N
H
iso-granulatimide A iso-granulatimide B iso-granulatimide C
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Pyrrolo-[3-Carbazole Derivatives.
Compound "a", below, represents a typical intermediate in the synthesis of
certain disclosed
pyrrolo-(3-carboline compounds, and was reported by Davis et al. ( J. Med.
Chem., 1992, 35, 177) as
an inhibitor of PKC. This compound was prepared using a different chemistry
than that of the instant
invention, and has not been further elaborated or cyclized.
A
Compound " b", above, has been proposed as an undesirable, unstable
intermediate in the
synthesis of staurosporine aglycone. The compound was not isolated or further
elaborated (Wood, J.
L. et al., J. Am. Claem. Soc., 1997,119, 9641).
A variety of synthetic procedures have been reported in the literature for the
preparation of 3-
(1H indol-3-yl)-lHpyrrole-2,5-diones involving the condensation of indole with
maleimide
(Bergman, J. et al. Tet~ahedt°on, 1999, 55). Most of these synthetic
procedures have distinct
limitations with regards to the use of harsh reaction conditions, thereby
limiting the functional groups
tolerated during the coupling reactions, the need for protection of the,
pyrrole-2,5-dione nitrogen, and
the total number of synthetic steps required for the preparation of the
desired indolocarbazole nuclei.
SUMMARY OF THE INVENTION
The present invention provides novel pyrrolo-(3-carboline derivatives and ring-
substitution
and structural derivatives of 3-(1H indol-3-yl)-1H pyrrole-2,5-diones.
Compounds disclosed herein
are useful for the treatment of neurodegenerative diseases, facilitating
regulation of the IAPs,
inhibition of various serine-threonine protein kinases, inhibiting the
degradation, dysfunction, or loss
of neurons of the CNS and PNS, or enhancing the phenotype of neuronal cells
and neuronal progress
and development, either in the CNS or in the PNS.
Compounds disclosed herein are also.useful in the prevention and treatment of
other disorders
and physiological conditions characterized by loss of growth and cellular
differentiation control, as
exemplified in cancer and inflammation, and various human and viral signal
transduction processes.
This utility arises from the inhibition of various protein kinases or by the
down regulation of the IAPs
including, but not limited to HIAP1,~HIAP2, and XIAP. Downregulation of anti-
apoptotic genes in
cancer cell lines, causing cell death, is useful in cancer chemotherapy.
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The invention also relates to a general synthetic route, permitting
preparation of pyrrolo-[3-
carboline derivatives (Structure II) which are distinct from the
indolocarbazole class of compounds
and their synthetic precursors, which represent 3-(indol-3-yl)-4-(1-aza-
heterosubstituted)-1H pyrrole-
2,5-diones (Structure I). Various substituted 3-(indol-3-yl)-1H pyrrole-2,5-
diones can be prepared
using a related, one-pot, chemical procedure. Selected compounds of this class
display potent
biological activity. For example, these compounds display ih vitYO
neuroprotection against multiple
apoptotic stresses. These anti-apoptotic compounds are usefull in the
treatment of acute and chronic
neurodegeneration, which has been further exemplified in animal models of PD.
Also included are inventive methods for the preparation of the compounds
disclosed herein.
The ring-substitution pyrrolo-(3-carboline derivatives and precursors of the
present invention
are compounds of formula I and II:
A2' \ N / 'B2 A2I \ N / 'B2
RvX3~ X4 ~N X9 X8. R$ RvX3~ X4 ~N X9 X8~ R$
'2 O ~ < ~O I 7 '2 0 ~ ~ ~~ I 7
R2~X~ ~ ~ s 6.XwR7 R2~X~ ~ a . 5 6,XwR7
R~ R~ R5 . R6 R~ R4 R5 R6
I II
Formulas I and II have functional groups designated as Al, A2, Bl, BZ, Xl-Xs
and R'-R8 as
defined further herein. The difference between formula I and formula II
resides in the bonds made
with the carbon atom shown above as "a" in formula II. The dashed line in
formula II indicates that
the bond between carbon "a" and XS may be either single or double bond. The
definitions of
functional groups having the same designations are the same for compounds of
formula I and II, but
may differ from functional groups having the same designation in formula III
(below).
The 3-(indol-3-yl)-1H pyrrole-2,5-diones analogues of the present invention
are compounds
of formula III:
H
A2 ~B2
R ~X~
2_X? ~ ~~Rs
R X3 N
III
Formula III has functional groups designated as A', Az, B1, BZ, X'-X3, R'-R5,
and Y as
defined further herein. The definitions of functional groups for compounds of
formula III may differ
from the definitions of functional groups having the same designation with
respect to formula I and II.
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DETAILED DESCRIPTION
Ring-Substitution and Structural Derivatives of Pyrrolo-(3-Carboline
Derivatives
Disclosed herein are pharmaceutically active ring-substitution and structural
derivatives of
pyrrolo-~3-carbolirle derivatives, represented by formulas I and II:
A2' \ N / _B2 ~ A2' \ N / _B2
R ~ X3~ X4 ~ ~N X9 Xg. R$ R3~ X3~ X4 ~N X9 Xg~ R$
n v ,
R2~ X~~ > \ 5 ~ Xs R7 R2~ X~~ a , 5 ~ XW R7
I II
and pharmaceutically acceptable salts thereof wherein:
formula I represents the non-cyclized form of formula II, and formula II is
defined as either
having a single or double bond between carbon "a" and X5;
A1 is H or lower alkyl, AZ is H, ORz°, or SRz°, having S' or R
stereochemistry, wherein Rzo
represents H, lower alkyl, aryl, substituted aryl, heteroaryl, or substituted
heteroaryl; or A' and AZ are
combined to represent oxygen;
B1 is H or lower alkyl, and BZ is H, ORZ°, or SRZ°, having S or
R stereochemistry; or B1 and
BZ are combined to represent oxygen;
Xl - X3 are independently C or N;
X4 is CH or N, wherein not more than two of X' - X4 is N;
XS represents N, C, or S when bound to carbon "a" with a double bond, and XS
represents CH
or N when bound to carbon "a" with a single bond;
X6 - X$ are independently C or N;
X9 is CH or N, wherein not more than two of X6 - X9 is N;
~ Rl - R3 and R6 - R8 represent a lone pair or O when each respective Xl- X3
and X6 - X$ is N;
and
when Xl - X3 or X6 - X$ is C, each respective Rl - R3 and R6 - R8 is
independently selected
from the group consisting of:
a H, lower all 1 lower substituted al 1 hi her al l, halo en, azido c ano
nitro or NRZIRza
eY~ kY~ g k3' g ~ Y > > >
wherein R21 represents H or lower alkyl, and R2z represents H, lower allcyl,
acyl, formyl, lower
alkylcarbonyl, substituted lower alkylcarbonyl, arylcarbonyl, substituted
arylcarbonyl,
heteroarylcarbonyl, substituted heteroarylcarbonyl carbamoyl, lower
allcylaminocarbonyl,
arylaminocarbonyl, or substituted arylaminocarbonyl; '
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b) ORz3, wherein Rz3 is H, acyl, lower alkylcarbonyl, lower alkylcarbonyl,
substituted lower
alkylcarbonyl, arylcarbonyl, substituted arylcarbonyl, alkylcarbamoyl,
arylcarbamoyl,
substituted arylcarbamoyl, heteroarylcarbamoyl, substituted heterocarbamoyl;
C) SRz3;
d) O(CHz)~-Rz4, O(CHz)~-O-Rz4, or O(CHz)~-S-Rz4, wherein j is an integer from
1 to 8, and Rz4 is
selected from the group consisting of H, lower alkyl, substituted lower alkyl,
aryl, substituted
aryl, heteroaryl, or substituted heteroaryl;
e) S(CHz)~Rz4, S(CHz)~-O-Rz4, or S(CHz)~-S-Rz4;
f) C=C-RzS, C---C-ORzs, or C=C-COZRzs, wherein Rz5 is H, lower alkyl,
substituted alkyl, aryl,
substituted aryl, heteroaryl, or substituted heteroaryl;
g) CH=CH-RzS, CH=CH-ORzs, or CH=CH-C02Rz5, having a stereochemistry of E or Z;
h) C---C-NRzSRzs or C=CCONRz5Rz6, wherein Rz6 is defined as Rzs, and Rzs and
Rz6 are selected
independently;
i) CH=CH-NRzSRzs or CH=CHCONRz5Rz6, having a stereochemistry of E or Z;
j) (CHz)kR2s, (CHz)k-COORzs, or (CHz)k-ORzs, wherein k is an integer from 2 to
6;
1e) (CHz)kNRz5Rz6, (CHz)kCONRz5Rz6, wherein Rzs and Rz6 are selected
independently; and
1) CHzXRz', wherein X is O or S and Rz' is H, lowed alkyl or substituted lower
alkyl, aryl, substituted
aryl, heteroaryl, substituted heteroaryl;
R4 is selected from the group consisting of:
m) H, lower alkyl, substituted lower alkyl, lower allcylcarbonyl, substituted
lower allcylcarbonyl,
arylcarbonyl, substituted arylcarbonyl, heteroarylcarbonyl, or substituted
heteroarylcarbonyl;
n) (CHz)kRzB, (CHz)k-COORzB, wherein k is an integer from 1 to 6, and Rz$ is
defined as H, lower
alkyl, aryl, substituted aryl, heteroaryl,or substituted heteroaryl;
o) (CHz)mXRz9 wherein m is an integer from 1 to 8, X is either O or S, and Rz9
is H, lower alkyl,
substituted lower alkyl, acyl, lower alkylcarbonyl, arylcarbonyl, substituted
arylcarbonyl, aryl,
substituted aryl, CHz-substituted aryl, heterocycle, CHz-substituted
heterocycle, or an a- or ~3-
antipoid sugar moiety;
p) (CHz)mNR3°R31 wherein m is an integer from 1 to 8, and R3°
and R31 are independently defined as
Rz9 above, or wherein R3° and R31 together are part of a heteroalkyl,
substituted heteroalkyl,
heteroaryl, or substituted heteroaryl ring system;
c~ (CHz)mXCONHR3z wherein m is an integer from 1 to 8, X is either O, S, or
NH, and R3z is
defined as Rz9;
r) CHzCH(OR33)CH20R34, wherein each R33 and R34 are independently defined as
H, lower alkyl,
substituted lower alkyl, acyl, lower alkylaminocarbonyl, arylaminocarbonyl,
substituted
arylaminocarbonyl, aryl, CHz-substituted aryl, heterocycle, or CHz-substituted
heterocycle;
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s) CO(CHZ)nR3s, wherein n is an integer from 1 to 8, and R~5 is selected from
the group consisting of
H, halogen, aryl, substituted aryl, heterocycle, unsubstituted heterocycle,
COR36, wherein R36 is
selected from the group consisting of H, lower alkyl, substituted lower alkyl,
acyl, carbamoyl,
lower alkylaminocarbonyl, arylaminocarbonyl, substituted arylaminocarbonyl,
aryl, CHZ-
substituted aryl, heterocycle, and CHZ-substituted heterocycle, and CONR3'R38,
wherein R3'
and R38 are independently selected from R29, or R3' and R3$ together comprise
a heteroalkyl,
substituted heteroallcyl, heteroaryl, or substituted heteroaryl ring system;
t) CO(CHa)nXR39, wherein n is an integer from 1 to 8, X is selected from O and
S, and R39 is defined
aS R36;
u) CO-sugar, wherein said sugar comprises 1 to 5 a- or Vii- antipoid sugar
moieties, or a combination
of substituted a- or (3- antipoid sugar moieties;
v) COCRø°R41R42, wherein R4° is H or lower alkyl, and where R41
and R4z together comprise a
substituted alkyl or substituted heteroalkyl ring system;
w) SOZR43, wherein R43 is selected from the group consisting of hydroxyl,
aryl, substituted aryl,
heteroaryl, substituted heteroaryl, (CHZ)pH, (CHZ)pOH, and (CHZ)pR44, wherein
p is an integer
from 1 to 8, and Rø4 is either OR45 , wherein R45 is defined as R29, or
NR~6R4', wherein R46 and
R4' are independently selected from the group consisting of H, lower alkyl,
substituted lower
alkyl, acyl, lower alkylcarbonyl, arylcarbonyl, substituted arylcarbonyl,
aryl, CHz-substituted
aryl, heterocycle, CHZ-substituted heterocycle, and an a- or (3-antipoid sugar
moiety;
x) a sugar comprising from 1 to 5 a- or (3- antipoid sugar moieties, or a
combination of substituted a-
or (3- antipoid sugar moieties; and
y) a polypeptide chain of between 1 and 10 amino acids, comprising protected
or unprotected D- or L-
amino acids, being attached to carbazole nitrogen at the carboxy terminus of
the polypeptide
chain;
RS is selected from the group consisting of
z) a lone pair when XS is S or N; and
aa) when XS is C, a substitution pattern according to any one of a) through
y); or
R4 and RS together are part of a substituted or unsubstituted alkyl, allcenyl,
heteroallcyl or
heteroalkenyl ring system, said ring system having from 5 to 7 ring members in
formula II and 7-9
ring members in formula I, a heteroatom of said ring system being selected
from N, O or S;
substitution patterns of said ring system being selected from the group
consisting of a) through y),
wherein at least one carbon of said ring system is unsubstituted; and
wherein in formula I, when A1 and AZ , and Bl and B2 , respectively combine to
form oxygen,
Rl-R3 and RS-R$ are H, and R4 is H or CH3, at least one of Xl - X9 represents
a ring member other
than carbon.
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Notably, within the structure of formula I or II; up to two of the outer ring
members of either
indole benzene rings, at positions Xl to X4 and X6 to X9 may be N. Thus, each
of the two
indole/indoline benzene rings may have zero, one or two N present at any of
the outer four positions.
Ring Substitution and Derivatives of 3-(1H Indol-3-yl)-1H Pyrrole-2,5-Dione
Disclosed herein are pharmaceutically active ring-substitution and structural
derivatives of 3-
(indol-3-yl)-1H pyrrole-2,5-diones represented by formula III:
H
A1 N B1
A2 ~B2
1
R ~X1,
2_X? ~ ~>--R5
R X3 N
R3 R4
or a pharmaceutically acceptable salt thereof wherein:
10 A1 is H or lower alkyl, Az is H, OR2°, or SRZ°, having S or R
stereochemistry, wherein RZo
represents H, lower alkyl, aryl, substituted aryl, heteroaryl, or substituted
heteroaryl; or A' and AZ are
combined to represent oxygen;
Bi is H or lower allcyl, and BZ is H, ORz°, ox SRZ°, having S or
R stereochemistiy; or B1 and
BZ are combined to represent oxygen or sulfur;
Xl - X3 are independently C or N; wherein not more than two of X', XZ and X3
is N;
Y is hydrogen, halogen, hydroxide, or lower alkyl;
Rl , RZ, and R3 represent a lone pair or O when X', Xz and X3, respectively,
is N;
Rl, R2, R3 (when Xl, XZ and X3, are C, respectively) and RS are independently
selected from
the group consisting of:
a) H, lower alkyl, lower substituted alkyl, higher alkyl, halogen, nitro, or
NRZ'R'z, wherein Rz~
represents H or lower alkyl, and R2z represents H, lower alkyl, formyl, acyl,
lower
allcylcarbonyl, substituted lower allcylcarbonyl, arylcarbonyl, substituted
arylcarbonyl,
heteroarylcarbonyl, substituted heteroarylcarbonyl carbamoyl, lower
alkylaminocarbonyl,
arylaminocarbonyl, or substituted arylaminocarbonyl;
b) ORz3, wherein R23 is H, acyl, lower allcylcarbonyl, lower alkylcarbonyl,
substituted lower
alkylcarbonyl, arylcarbonyl, substituted arylcarbonyl, alkylcarbamoyl,
arylcarbamoyl,
substituted arylcarbamoyl, heteroarylcarbamoyl, substituted heterocarbamoyl;
C) SR23;
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d) O(CHz)~-Rzø, O(CHz);-O-Rz4, or O(CHz)~-S-Rz4, wherein j is an integer from
1 to 8, and Rz4 is
selected from the group consisting of H, lower alkyl, substituted lower alkyl,
aryl, substituted
aryl, heteroaryl, or substituted heteroaryl;
e) S(CHz)~RZa, S(CHz)~-O-Rz4, or S(CHz)~-S-Rz4;
f) C=C-Rzs, C=C-ORzS, or C=C-COZRzs, wherein Rz$ is H, lower alkyl,
substituted alkyl, aryl,
substituted aryl, heteroaryl, or substituted heteroaryl;
g) CH=CH-Rzs, CH=CH-ORzs, or CH=CH-COZRzS, having a stereochemistry of E or Z;
h) C---C-NRz5Rz6 or C---CCONRz5Rz6, wherein Rz6 is defined as Rzs, and Rzs and
Rz6 are selected
independently;
i) CH=CH-NRz5Rz6 or CH=CHCONRzSRzs, having a stereochemistry of E or Z;
j) (CHz)kRzs, (CHz)k-COORzs, or (CHz)k=ORzS, wherein k is an integer from 2 to
6;
k) (CHz)kNRz5R2s, (CHz)kCONRz5Rz6, wherein Rzs and Rz6 are selected
independently; and
1) CHzXRz', wherein X is O or S, and Rz' is H, lowed alkyl or substituted
lower alkyl, aryl, substituted
aryl, heteroaryl, substituted heteroaryl;
R4 is selected from the group consisting of:
m) H, lower alkyl, substituted lower alkyl, lower alkylcarbonyl, substituted
lower alkylcarbonyl,
arylcarbonyl, substituted arylcarbonyl, heteroarylcarbonyl, or substituted
heteroarylcarbonyl;
n) (CHz)kRzB, (CHz)k-COORzs, wherein lc is an integer from 1 to 6, and Rz8 is
defined as H, lower
alkyl, aryl, substituted aryl, heteroaryl,or substituted heteroaryl;
0) (CHz)mXRz9 wherein m is an integer from 1 to 8, X is either O or S, and Rz9
is H, lower alkyl,
substituted lower alkyl, acyl, lower alkylcarbonyl, arylcarbonyl, substituted
arylcarbonyl, aryl,
substituted aryl, CHz-substituted aryl, heterocycle, CHz-substituted
heterocycle, or an oc- or (3-
antipoid sugar moiety;
p) (CHz)mNR3°R3' wherein m is an integer from 1 to 8, and R3°
and R3' are independently defined as
Rz9 above, or wherein R3° and R3' together are part of a heteroallcyl,
substituted heteroallcyl,
heteroaryl, or substituted heteroaryl ring system;
~ (CHz)mXCONHR3z wherein m is an integer from 1 to 8, X is either O, S, or NH,
and R3z is
defined as Rz9;
r) CHZCH(OR33)CHaOR34, wherein each R33 and R34 are independently defined as
H, lower alkyl,
substituted lower alkyl, acyl, lower alkylaminocarbonyl, arylaminocarbonyl,
substituted
arylaminocarbonyl, aryl, CHz-substituted aryl, heterocycle, or CHz-substituted
heterocycle;
s) CO(CHZ)nR3s, wherein n is an integer from 1 to 8, and R35 is selected from
the group consisting of
H, halogen, aryl, substituted aryl, heterocycle, unsubstituted heterocycle,
COR36, wherein R3g is
selected from the group consisting of H, lower alkyl, substituted lower alkyl,
acyl, carbamoyl,
lower alkylaminocarbonyl, arylarr~inocarbonyl, substituted arylaminocarbonyl,
aryl, CHz-
substituted aryl, heterocycle, and CHz-substituted heterocycle, and CONR3'R38,
wherein R3'
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and R3$ are independently selected from R29, or R3' and R3g together comprise
a heteroalkyl,
substituted heteroalkyl, heteroaryl, or substituted heteroaryl ring system;
t) CO(CH2)nXR39, wherein n is an integer from 1 to 8, X is selected from O and
S, and R39 is defined
as R36;
u) CO-sugar, wherein said sugar comprises 1 to 5 a- or (3- antipoid sugar
moieties, or a combination
of substituted a- or (3- antipoid sugar moieties;
v) COCR4°R4'R42, wherein R4° is H or lower allcyl, and where R4'
and R4z together comprise a
substituted alkyl or substituted heteroalkyl ring system;
w) SOZR43, wherein R43 is selected from the group consisting of hydroxyl,
aryl, substituted aryl,
heteroaryl, substituted heteroaryl, (CHZ)pH, (CHZ)pOH, and (CHZ)pR44, wherein
p is an integer
from 1 to 8, and R44 is either OR45 , wherein R45 is defined as Rz9, or
NR46R4', wherein R46 and
R4' are independently selected from the group consisting of H, lower alkyl,
substituted lower
alkyl, acyl, lower alkylcarbonyl, arylcarbonyl, substituted arylcarbonyl,
aryl, CHI-substituted
aryl, heterocycle, CHZ-substituted heterocycle, and an a- or [3-antipoid sugar
moiety;
x) a sugar comprising from 1 to 5 a- or (3- antipoid sugar moieties, or a
combination of substituted a-
or [3- antipoid sugar moieties; and
y) a polypeptide chain of between 1 and 10 amino acids, comprising protected
or unprotected D- or L-
amino acids, attached to carbazole nitrogen at the carboxy terminus of the
polypeptide chain.
Notably, within the structure of formula III, any of the outer three positions
of the indole
benzene (denoted as X' to X3) may be C or N. Thus, the indole benzene may have
zero, one or two N
present at any of these three positions. The compounds represented by formula
I, II, or III are
hereinafter interchangeably referred to as Compound I, II or III,
respectively.
Pharmaceutically Acceptable Salts
Pharmaceutically acceptable salts of formula I, II, and III may be any salt
such as an acid salt,
a basic salt or a neutral salt. For example, a salt may be prepared by the
direct protonation of a
nitrogen found at any of positions X'-X9 in formulas I and II, or Xl-X3 in
formula III with a
pharmaceutically acceptable acid; or by the protonation of a basic nitrogen
found at any of positions
Rl-R9 of formula I and II, or Rl-RS or Y of formula III, with a
pharmaceutically acceptable acid.
These basic nitrogens are exemplified by primary, secondary, or tertiary
amines, and heteroaryl
moieties containing nitrogen, exemplified by pyridyl and quinolinyl ring
systems.
Pharmaceutically acceptable salts of formula I, II, and III may be prepared by
the treatment of
an acidic moiety found in a position such as Rl-Rg of formula I and II, or Rl-
RS or Y of formula III,
with a pharmaceutically acceptable base. These acidic moieties are exemplified
by carboxylic,
sulfonic, and boronic acids.
Pharmaceutically acceptable acid and basic salts are exemplified herein.
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Substitutent Definitions
In the definitions of the functional groups of formula I, II, and III, lower
alkyl means a
straight-chain or branched alkyl group having 1 to 8 carbon atoms, such as
methyl, ethyl, propyl, iso-
propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, pentyl, iso-amyl,
neopentyl, 1-ethylpropyl, hexyl, and
octyl. The lower alkyl moiety of lower alkoxy, lower alkylsulfonyl, lower
alkoxylcarbonyl, lower
allcylaminocarbonyl has the same meaning as lower alkyl defined above. The
acyl moiety of the acyl
and the acyloxy group means a straight-chain or branched alkanoyl group having
1 to 6 carbon atoms,
such as formyl, acetyl, propanoyl, butyryl, valeryl, pivaloyl and hexanoyl,
and arylcarbonyl group
described below, or a heteroarylcarbonyl group described below. The aryl
rizoiety of the aryl, the
arylcarbonyl and arylaminocarbonyl groups means a group having 6 to 12 carbon
atoms such as
phenyl, biphenyl, or naphthyl. The heteroaryl moiety of the heteroarylcarbonyl
groups contain at least
one hetero atom selected from O, N, and S, and includes pyridyl, pyrimidyl,
pyrroleyl, furyl, thienyl,
imidazolyl, triazolyl, quinolyl, iso-quinolyl, benzoimidazolyl, thiazolyl, and
benzothiazolyl. The
aralkyl moiety of the aralkyl and the aralkyloxy groups having 7 to 15 carbon
atoms, such as benzyl,
phenethyl, benzhydryl, and naphthylmethyl. The substituted lower alkyl group
has 1 to 3
independently selected substitutuents, such as hydroxyl, lower alkyloxy,
carboxyl, lower
allcylcarbonyl, nitro, amino, mono- or di-lower alkylamino, dioxolane,
dioxane, dithiolane, and
dithione. The lower alkyl moiety of the substituted lower alkyl, and the lower
alkyl moeity of the
lower alkoxy, the lower allcoxycarbonyl, and the mono- and di-lower
allrylamino in the substituents of
the substituted lower alkyl group have the same meaning as lower alkyl defined
above. By CHZ-
substituted, it is meant that the substituent is present on a CHZ carbon.
As used herein, the following terms denote functional groups: Me = methyl, Bn
= benzyl, Ph
= phenyl, tBu = t-butyl, Ac = acetyl, Ts = tosyl, pyr = pyruvate, Phth =
phthalate, Et = ethyl, Boc =
tent-butoxycarbonyl, Th = thiazyl, dansyl = 1-(5-(dimethylamino)napthyl), cat
= catalytic amount,
DMAP = dimethylaminoaminopyridine, Piv = pivavoyl, Pf = pentafluorophenyl, and
DIC =
diisopropylcarbodiimide.
The substituted aryl, the substituted heteroaryl and the substituted aralleyl
groups each may
have from 1 to 3 independently-selected substitutents, such as lower alkyl,
hydroxy, lower allcoxy,
carboxy, lower allcoxycarbonyl, nitro, amino, mono or di-lower alkylarnino,
and halogen. The lower
alkyl moiety of the lower alkyl, the lower allcoxy, the lower allcylamino, and
the mono- and di-lower
alkylamino groups among the susbtituents has the same meaning as lower alkyl
defined above. The
heterocyclic group formed with a nitrogen atom includes pyrroleyl,
piperidinyl, piperidino,
morpholinyl, morpholino, thiomorpholino, N-methylpiperazinyl, indolyl, and
isoindolyl. The
cycloalkyl moeity means a cycloalkyl group of the indicated number of carbon
atoms, containing one
or more rings anywhere in the structure, such as cycloalkyl groups include
cyclopropyl,
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cyclopropylmethyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-norbornyl, and 1-
adamantyl. The lower
fluoroalkyl moiety means a lower fluoroalkyl group in which one or more
hydrogens of the
corresponding lower alkyl group, as defined above, is replaced by a fluorine
atom, such as CHZF,
CHFz, CF3, CHZCF3. The a-amino acids include alanine, aminobutyric acid,
arginine, asparagine,
aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine,
isoleucine, leucine, lysine,
methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine,
valine, as well as other
amino acids which may occur naturally, or which can be derived from naturally
occurring amino
acids. The amino acids may be in the L-form, or the D-form, or in the form of
racemates. The
polypeptide groups include any linear combination of the above a-amino acids.
Halogen includes
fluorine, chlorine, bromine, and iodine.
Some of the compounds described herein contain one or more chiral centres and
may thus
give rise to diastereomers and optical isomers. The present invention is meant
to comprehend such
diastereomers as well as their racemic, resolved and enantiomerically pure
forms, and
pharmaceutically acceptable salts thereof.
Some of the compounds described herein contain olefinic double bonds, and
unless specified
otherwise, are meant to include both E and Z isomers.
Neuroprotective Profile of Compounds According to Formulas I, II, and III
The following section summarizes the biological profiles of the compounds
defined as
formula I through III, in cultured CNS-derived cerebral granule neurons (CGN),
neuroblastoma cell
lines SHSY-SY and LANS, and cortical cell lines, treated with various~pro-
apoptotic triggers.
The compounds defined by formula I through III protect CGNs against several
pro-apoptotic
triggers, including high/low potassium (HK/L,K), (3-amyloid (A(3) fibril
formation, ceramide,
glutamate, and cisplatin. Additionally, these compounds down regulate the
dramatic increase in
caspase induction observed during IiK/LK treatments, suggesting they prevent
cell death by
interfering in the apoptotic cascade at a point upstream of the caspases, ie,
the inhibition of one or
several of the serine/threonine protein kinases directly upstream of the
caspases, typified by MEKK1,
MKL, JNK, and P53.
Cultured CGNs which are maintained in a medium containing 26 mM potassium
(high K+ or
HK) undergo cell death when the medium is changed to one containing 5 mM
potassium (low K+ or
LK). HK maintains the cells in a highly polarized state, duplicating that of
fully innervated neurons.
The switch to LK (5 mM is more representative of physiological conditions)
results in depolarization
of the cells, mimicking the loss of neuronal conductivity. Cell death under
these conditions displays
typical features of apoptotic cell death, both in morphology and in the
upregulation of various killer
genes including c jun and the caspases 1 and 3 (Ikeuchi, T, Hurn. Cell,1998,
11, 125). As these killer
genes are turned on by various types of neuronal insult as observed in AD, PD,
and stroke, HK/LK is
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a general in vitf°o model for neuronal degradation, blanketing a wide
range of neurodegenerative
diseases. Compounds that protect against HK/LK in CGNs would therefore be
efficacious in
treatment and/or prevention of various neurodegenerative disease states.
The inventive compounds inhibit HK/LK apoptotic cell death in.CGNs with
selected
5 compounds protecting upwards of 100% of the neurons at 10 ~,m drug
concentrations with ICso values
in the range of 1-10 ~M (see Example 112). K252a and CEP 1347 displayed ICso
values of 0.3 and 1
~M, respectively. These compounds, however, were toxic at higher doses, while
the compounds
listed in Example 112 displayed little or no toxicity in untreated controls.
Caspase 3 expression is potently induced during in vitro HK/LK lcilling of
CGNs. At
10 concentrations of 10 ~M (which corresponds to 75-100 % protection of CGN
neurons), compounds 42
and 64 significantly inhibited caspase 3 induction by 50 % and 33 %,
respectively. Caspase 3
induction ultimately leads to cell death and is observed in any number of
neurodegenerative diseases.
Compounds which inhibit caspase 3 induction represent potent therapies for
various
neurodegenerative diseases.
15 Extracellular A(3 fibril formation has been found to be toxic to neurons,
and represents one
trigger for apoptotic death in AD. Various mechanisms have been put forward in
order to account for
the neurotoxicity related to extracellular A(3 fibril formation. Some of these
include altered enzyme
activity and disrupted calcium homeostasis leading to calpain and caspase
activation (Chan, S. L,
Mattson, M. P. J. Neu~osci. Res., 1999, 58, 167), increased free radical
formation, and more recently,
A(3 has been shown to interact with various receptor sites and to physically
insert into the cell
membrane (Kanfer, J: N. et al. Neuroclaern Res., 1999, 24, 1621). Regardless
of the mode of action,
extracellular A(3 fibril formation acts as an effective apoptotic trigger for
neuronal cells and serves as
an in vitro model for various neurodegenerative diseases characterized by
extracellular protein fibril
formation, typified by diseases such as AD and PD.
Linear A(3 rapidly aggregates in CGN cultures, leading to the apoptotic cell
death of
approximately 50% of the neurons after 5 days. Addition of selected compounds
of the formula I
through III saves upwards of 100% of these cells at drug concentrations of 10
~M with IC~o values of
1-10 ~,M (Example 113). These compounds may be added to the CGN culture 24
hours prior to linear
A(3 addition or at the time of linear A(3 addition. Similar saves were
observed under these two
scenarios. The most active compounds displayed limited toxicity, less than 5%,
in their respective in
vitro controls. As observed in the HK/LK assays, CEP 1347 displayed limited
protection (> 10 % at
concentrations below 300 nMJ and severe toxicity at concentrations greater
than 300 nM.
Ceramide is a native protein found in most mammalian and human cells. The
upregulation of
endogenous ceramide has been linked to caspase 1 (ICE) induced apoptosis
(Suzuki, A. et al. Exp.
Gel. Res., 1997, 233, 41). In a similar fashion, the addition of ceramide to
cultured CGNs results in
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16
caspase induced apoptosis, and is therefore considered an effective irz vitro
model for the various
neurodegenerative diseases described above, which are characterized by caspase
induced apoptosis, as
observed in stroke models. Addition of selected compounds of the formula I
through III, 24 hours
pxior to the addition of ceramide, to cultured CGNs provided protection
against apoptosis, with 10 to
20% of the cells being saved at 1-10 ~M drug concentrations (see Example 114).
Glutaminergic neurons secrete the neurotransmitter glutamate as a part of
normal cell
signaling processes. Intracellular glutamate levels are regulated by glial
cell uptake and conversion to
glutamine. Under conditions of oxidative stress, as observed in various
neurodegenerative diseases
such as stroke, ALS, PD, and HD, glutaminergic neurons release massive amounts
of glutamate into
the intracellular fluid, overwhelming the surrounding cells. Stimulation of
both NMDA and non-
NMDA-type glutamate excitory receptors leads to sustained depolarization of
postsynaptic
dendrosomal membranes, increased membrane permeability, and impaired ion
homeostasis, all
leading to either apoptotic or necrotic cell death.
Addition of selected compounds of the formula I through III, either at the
time of glutamate
addition or 24 hours prior to the addition of ceramide, to cultured CGNs
provided protection against
neuronal cell death, with up to 20% of the cells being saved at 1-10 p,M drug
concentrations (Example
115).
Cisplatin has been used extensively in the treatment of various cancers. One
dose-limiting
side effect of this chemotherapeutic agent is related to hearing loss as a
result of its toxicity to
auditory neurons and loss of feeling in the extremities. Cisplatin induces
apoptosis to both cultured
CGN and cortical neurons. Melatonin has been shown to protect auditory neurons
during cisplatin
treatments ifa vivo, and this combination therapy is currently in clinical
trial. We have shown that at
high doses melatonin will also protect CGNs, suggesting that protection of
CGNs may serve as an in
vitro model for cisplatin induced neuropathies.
Addition of selected compounds of the formula I through III, 24 hours prior to
the addition of
cisplatin (25 ~g/mL), to cultured CGNs protected against neuronal apoptosis
(see Example 116).
Compounds 35 and 36 displayed ICSO of 100 nM (80 % survival at 300 nM) against
cisplatin induced
apoptosis in CGNs. Several other compounds of the formula I through III
displayed ICso values in the
range of 3-10 ~tM, with upwards of 80 % neuronal survival. In contrast CEP
1347 displayed limited
protection (> 30%) at 1 p,M.
Cisplatin also induces apoptosis in primary cortical neurons. Compounds 51 and
52 protected
20 and 40%, respectively, of the cultured cortical neurons at concentrations
of 10 p.M. Compounds 35
and 36.protected 10 % of neurons. CEP 1347 protected 35% ofthese neurons at
0.3 pM. These
compounds represent novel therapies for the prevention of neuronal damage
caused by DNA
damaging chemotherapeutic agents (see Example 117).
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Cultured Superiour Cervical Ganglion (SCG) neurons of the PNS which have been
sustained
in media containing Neuron Growth Factor (NGF) undergo apoptotic cell death
upon removal of the
NGF from the cellular medium. Loss of neuronal trofm support has been
implicated in various
neurodegenerative disease states in the PNS, such as diabetic neuropathy and
neuropahies related to
chemotherapeutic and anti-HIV drug treatment.
Compounds of the formula I through III protect SCG neurons against NGF
withdrawal when
applied at concentration of 10-20 pM (see Example 118). The disclosed
compounds are therefore
useful in the treatment of various periferal neuropathies.
Anti-Cancer Profile of Compounds Defined by Formulas I, II, and 11I
Regardless of its neuroprotective capability, if a compound interferes with
chemotherapy by
protecting cancer cells from an apoptotic stimuli then the compound is of less
value as an anti-cancer
therapeutic. Compound 36 according to the invention displayed good protection
of CGNs to cisplatin
induced apoptosis (ICso = 100 nM). Additionally, this compound did not protect
cisplatin sensitive
OV 2008 ovarian cancer cells from cisplatin (20 pg/mL) induced apoptosis.
Etoposide is a well known chemotherapeutic which is toxic to several neuronal
cell lines,
including CGNs. Etoposide is a topoisomerase I inhibitor which induces
cellular apoptosis by the
inhibition of regular cell cycle progression and DNA fragmentation. As
discussed above, compounds
disclosed herein are neuroprotective to various apoptotic insults. Ideally,
these compounds will not
interfere with chemotherapeutic treatments by protect cancer cells from the
same insult.
SHSY-SY cells are members of a neuroblastoma cell line. When SHSY-SYs were
pretreated
for 24 hours with selected compounds of the formula I through III, followed by
etoposide, little or no
protection was observed (see Example 119). This is a very positive result as
etoposide, like cisplatin,
i's a widely used chemotherapeutic agent, and protection against etoposide
lcills in cancer cells would
be detrimental to chemotherapy. As various topoisomerase I inhibitors are
currently in clinical trial as
anti-cancer agents, it is clear that concurrent administration of selected
compounds of the formula I
through III with specific topoisomerase I inhibitors will not interfere with
the topoisomerase I induced
cell death in cancer cells.
HAIP 1 and 2 are members of the IAPs which we have shown to be involved in
apoptotic
regulation in various cancer cell lines. Down-regulation of these proteins,
both at the translational and
protein levels, presents a novel means of inducing apoptosis in cancer cells.
Selected compounds of the formula I to III down-regulate the endogenous levels
of HIAPl
mRNA found in the neuroblastomal cell line LANS. mRNA levels were observed to
drop by as much
as 80% after 24 hours treatments of the respective cell lines with selected
compounds of the formula I
to III, as compared to control. These compounds represent new
chemotherapeutics for treatment of
cancer.
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1S
The term "subject" or "patient" as used herein refers to any mammal including
humans,
primates, horses, cows, pigs, sheep, goats, dogs, cats and rodents.
The pharmaceutical compositions of the invention are administered to subjects
so as to deliver
the compound of formula I to III in an effective amount. An effective amount
means that amount
necessary to delay the onset of, inhibit the progression of, halt altogether
the onset or progression of,
or diagnose the particular condition or symptoms of the particular condition
being treated. In general,
an effective amount for treating a neurological disorder is that amount
necessary to affect any
symptom or indicator of the condition. In general, an effective amount for
treating cancer will be that
amount necessary to favorably affect mammalian cancer cell proliferation ira
situ. When administered
to a subject, effective amounts will depend, of course, on the particular
condition being treated; the
severity of the condition; individual patient parameters including age,
physical condition, size and
weight; concurrent treatment; frequency of treatment; and the mode of
administration. These factors
are well known to those of ordinary skill in the art and can be addressed with
no more than routine
experimentation. Advantageously, a maximum dose is used, that is, the highest
safe dose according to
sound medical judgment.
A variety of administration routes are available. The particular mode selected
will depend, of
course, upon the particular condition being treated, the particular compound
selected, the severity of .
the condition being treated, and the dosage required for therapeutic efficacy.
The methods of this
invention, generally speaking, may be practiced using any mode of
administration that is medically
acceptable, meaning any mode that produces effective levels of the compounds
of any of formulas I to
III without causing clinically unacceptable adverse effects. Such modes of
administration include oral,
rectal, sublingual, topical, nasal, transdermal, intradermal or parenteral
routes. The term "parenteral"
includes subcutaneous, intravenous, intramuscular, or infusion. Oral routes
are advantageous because
of the ease with which a subject can ingest an oral dosage form.
Dosage levels may be adjusted appropriately to achieve desired levels of a
compound of
formula I to III, either locally or systemically. Generally, a daily oral dose
of a compound of formula I
to III will be from about 0.01 mg/kg per day to 1000 mg/kg per day. Three
doses per day, each in the
range of about 1 to 1000 mg/m2 per day would be effective. In the event that
the response in a subject
is insufficient at such doses, even higher doses (or effective higher doses by
a different, more
localized delivery route) may be employed to the extent that a subject's
tolerance permits.
The compositions containing compounds according to the invention may
conveniently be
presented in unit dosage form and may be prepared by any of the methods well
known in the art of
pharmacy. All methods include the step of bringing the compounds of the
invention into association
with a carrier that constitutes one or more accessory ingredients. In general,
the compositions are
prepared by uniformly and intimately bringing the compounds into association
with a liquid carrier, a
finely divided solid carrier, or both, and then, if necessary, shaping the
product.
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Compositions suitable for oral administration may be presented as discrete
units such as
capsules, cachets, tablets, or lozenges, each containing a predetermined
amount of the compound of
formula I to III. Other compositions include suspensions in aqueous liquors or
non-aqueous liquids
such as a syrup, an elixir, or an emulsion.
Other delivery systems can include time-release, delayed release or sustained
release delivery
systems. Such systems can avoid repeated administrations of the compounds of
formula I to III,
thereby increasing convenience to the subject and the physician. Many types of
release delivery
systems are available and known to those of ordinary skill in the art. They
include polymer based
systems such as polylactic and polyglycolic acid, polyanhydrides and
polycaprolactone; nonpolymer
systems that are lipids including sterols such as cholesterol, cholesterol
esters and fatty acids or
neutral fats such as mono-, di- and triglycerides; hydrogel release systems;
silastic systems; peptide
based systems; wax coatings, compressed tablets using conventional binders and
excipients, partially
fused implants and the like. In addition, a pump-based hardware delivery
system can be used, some of
which are adapted for implantation.
A long-term sustained release implant also may be used. "Long-term" release,
as used herein,
means that the implant is constructed and arranged to deliver therapeutic
levels of the active
ingredient for at least 30 days, and preferably 60 days. Long-term sustained
release implants are well
known to those of ordinary skill in the art and include some of the release
systems described above.
Such implants can be particularly useful in treating solid tumors by placing
the implant near or
directly within the tumor, thereby affecting localized, high-doses of the
compounds of the invention.
When administered, the compositions according to the invention may contain
other
pharmaceutically acceptable components. Such compositions may contain salts,
buffering agents,
preservatives, compatible carriers, and optionally other therapeutic
ingredients. When used in
medicine the salts should be pharmaceutically acceptable, but non-
pharmaceutically acceptable salts
may be used in synthetic reactions to prepare pharmaceutically acceptable
salts therefrom, and are not
excluded from the scope of the invention. Such salts include, but are not
limited to, those prepared
from the following acids: hydrochloric, hydrobromic, sulphuric, nitric,
phosphoric, malefic, acetic,
salicylic, p-toluenesulfonic, tartaric, citric, methane sulfonic, formic,
malonic, succinic, naphthalene-
2-sulfonic, and benzene sulfonic. Also, pharmaceutically acceptable salts can
be prepared as alkaline
metal or alkaline earth salts, such as sodium, potassium or calcium salts.
Suitable buffering agents include: acetic acid and salts thereof (1-2% W/~;
citric acid and
salts thereof (1-3% W/V); and phosphoric acid and salts thereof (0.8-2% W/V),
as well as others
known in the art. Suitable preservatives include benzalkonium chloride (0.003-
0.03% W/V);
chlorobutanol (0.3-0.9% W/V); parabens (0.01-0.25% W/V) and thimerosal (0.004-
0.02% W/V), as
well as others known in the art. Suitable carriers are pharmaceutically
acceptable carriers. The term
pharmaceutically acceptable carrier means one or more compatible solid or
liquid filler, dilutents or
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encapsulating substances that are suitable for administration to a human or
other mammal. The term
"carrier" denotes an organic or inorganic ingredient, natural or synthetic,
with which the active
ingredient is combined to facilitate the application. The components of the
pharmaceutically
acceptable carrier are capable of being commingled with the molecules of the
compounds of formula I
5 to III of the present invention, and with each other, in a manner such that
there is no interaction which
would substantially impair the desired pharmaceutical efficacy. Carrier
formulations suitable for oral,
subcutaneous, intravenous, and intramuscular administration etc., are those
known in the art.
The compounds of the invention may be delivered with other therapeutic agents.
The
invention additionally includes co-administration of any of the compounds of
formula I to III with
10 other compounds known to be useful in treating neurodegenerative diseases,
typified by but not
limited to, acetylcholinesterase inhibitors for treatind AD such as tacrine,
doneprizil, and rivastigmin,
and L-dopa for treating PD.
Compounds of formulas I to III have been shown to protect CGNs and cortical
neurons to
cisplatin induced apoptosis. This protection extends to the protection of
other CNS and peripheral
15 neurons to various neurotoxiris and chemotherapeutic agents which induce
CNS and/or peripheral
neurotoxicity.
In the case of peripheral neuropathy induced by a toxic agent, the compounds I
through III are
delivered separately before, simultaneously with (ie. in the form of anti-
cancer coctails, as described
in further detail below), or after exposure to the toxic agent. Optionally,
compounds of formula I
20 through III and the neurotoxic or chemotherapeutic agent are each
administered at effective time
intervals, during an overlapping period of treatment in order to prevent or
restore at least a portion of
the neurofunction destroyed by the neurotoxic or chemotherapeutic agent. The
chemotherapetic agent
can be any causing neurotoxicity, such as dideoxyinosine, cisplatin,
etoposide, vincristine, or taxol.
By "toxic agent" or "neurotoxic agent" is meant a substance that through its
chemical action
injures, impairs, or inhibits the activity of a component of the nervous
system. The list of neurotoxic
agents that cause neuropathies is lengthy (see a list of neurotoxic agents
provided in Table 1). Such
neurotoxic agents include, but are not limited to, neoplastic agents such as
vincristine, vinblastine,
cisplatin, taxol, or dideoxy-compounds, eg., dideoxyinosine; alcohol; metals;
industrial toxins
involved in occupational or environmental exposure; contaminants in food or
medicinals; or over-
doses of vitamines or therapeutic drugs, eg. Antibiotics such as penicillin or
chloramphenicol, or
mega-doses of vitamins A, D, or B6.
Table 1- Neurotoxic Agents
AGENT . ACTIVITY AGENT ACTIVITY
actazolimide Diuretic imipramine antidepressant
acrylamide flocculant, grouting agent indolinethacin anti-inflammatory
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Table 1- Neurotoxic Agents
AGENT ACTIVITY AGENT ACTIVITY
adriamycin Antineoplastic inorganic leadtoxic metal in
paint, etc.
alcohol (ie. solvent, recreationaliso-niazid antituberculousis
ethanol) drug
alinitine respiratory stimulantlithium antidepressant
amiodarone Antiarrtliymic methylxnercuryindustrial waste
amphotericin Antimicrobial metformin antidiabetic
arsenic herbicide, insecticidemethylhydrazinesynthetic intermediate
aurothioglucose Antirheumatic metronidazole antiprotozoal
barbiturates anticonvulsive, misonidazole radiosensitizer
sedative
buckthorn toxic berry nitrofurantoinurinary antiseptic
carbimates Insecticide nitrogen mustardantineoplastic,
nerve gas
carbon disulfideindustrial applicationsnitous oxide anesthetic
chloramphenicol Antibacterial organophosphatesinsecticides
chloroquW a Antimalarial ospolot anticonvulsant
chlorestyramine Antihyperlipoproteinemicpenicillin antibacterial
cisplatin Antineoplastic perhexiline antiarrhytbmic
clioquinol amebicide, antibacterialperhexiline antiarrythmic
maleate
colestipol Antihyperlipoproteinemicphenytoin anticonvulsant
colchicine gout suppressant platnim drug component
colistin Antimicrobial primidone . anticonvulsant
cycloserine Antibacterial procarbazine antineoplastic
cytarabine Antineoplastic pyridoxine vitamin B6
dapsone dermatological sodium cyanateantisickling
ie- leprosy
dideoxycytidine Anatineoplastic streptomycin antimicrobial
dideoxyinosine Antineoplastic sulphonamides antimicrobial
dideoxythymidineAntiviral suramin anteneoplastic
disulfiram Antialcohol tamoxifen antineoplastic
doxorubicin Antineoplastic taxol antineoplastic
ethambutol Antibacterial thalidomide antileprous
ethionamide Antibacterial thallium rat poison
glutethimide sedative, hypnotictriamterene diuretic
gold . Antirheumatic trimethyltin toxic metal
hexacarbons Solvents L-trypophan health food additive
hormonal contraceptives vincristine Antineoplastic
hexamethylolxnelaminefireproofing, creasevinblastine Antineoplastic
proofing
hydralazine Antihypertensive vindesine Antineoplastic
hydroxychloroquineAntirheumatic vitamine A mega doses
or D
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In the case of cancer, the compounds would be delivered separately or in the
form of anti-
cancer cocktails. An anti-cancer cocktail is a mixture of any one of the
compounds having formula I
to III with another anti-cancer agent such as an anti-cancer drug, a cytokine,
and/or supplementary
potentiating agent(s). The use of cocktails in the treatment of cancer is
routine. In this embodiment, a
common administration vehicle (e.g., pill, tablet, implant, injectable
solution, etc.) could contain both
a compound of formula I to IlI and the anti-cancer drug and/or supplementary
potentiating agent.
Thus, cocktails comprising of formula I through III compounds as well as other
compounds
are within the scope of the invention. Compounds having anti-neoplastic
properties include, but are
not limited to: Antineoplastic: Acivicin; Aclarubicin; Acodazole
Hydrochloride; Acronine;
Adozelesin; Aldesleukin; Altretamine; Ambomycin; Ametantrone Acetate;
Aminoglutethimide;
Amsacrine; Anastrozole; Anthramycin; Asparaginase; Asperlin; Azacitidine;
Azetepa; Azotomycin;
Batimastat; Benzodepa; Bicalutamide; Bisantrene Hydrochloride; Bisnafide
Dimesylate; Bizelesin;
Bleomycin Sulfate; Brequinar Sodium; Bropirimine; Busulfan; Cactinomycin;
Calusterone;
1 S Caracemide; Carbetimer; Carboplatin; Carmustine; Carubicin Hydrochloride;
Carzelesin; Cedefingol;
Chlorambucil; Cirolemycin; Cisplatin; Cladribine; Crisnatol Mesylate;
Cyclophosphamide;
Cytarabine; Dacarbazine; Dactinomycin; Daunorubicin Hydrochloride; Decitabine;
Dexormaplatin;
Dezaguanin; Dezaguanine Mesylate; Diaziquone; Docetaxel; Doxorubicin;
Doxorubicin
Hydrochloride; Droloxifene; Droloxifene Citrate; Dromostanolone Propionate;
Duazomycin;
Edatrexate; Eflornithine Hydrochloride ; Elsamitrucin; Enloplatin; Enpromate;
Epipropidine;
Epirubicin Hydrochloride; Erbulozole; Esorubicin Hydrochloride; Estramustine;
Estramustine
Phosphate Sodium; Etanidazole; Ethiodized Oil I 131; Etoposide; Etoposide
Phosphate; Etoprine;
Fadrozole Hydrochloride; Fazarabine; Fenretinide; Floxuridine; Fludarabine
Phosphate; Fluorouracil;
Flurocitabine; Fosquidone; Fostriecin Sodium; Gemcitabine; Gemcitabine
Hydrochloride; Gold Au
198; Hydroxyurea; Idarubicin Hydrochloride; Ifosfamide; lmofosine; Interferon
Alfa-2a; Interferon
Alfa-2b ; Interferon Alfa-nl; Interferon Alfa-n3; Interferon Beta-Ia;
Interferon Gamma-Ib; Iproplatin;
Irinotecan Hydrochloride; Lanreotide Acetate; Letrozole; Leuprolide Acetate;
Liarozole
Hydrochloride; Lometrexol Sodium; Lomustine; Losoxantrone Hydrochloride;
Masoprocol;
Maytansine; Mechlorethamine Hydrochloride; Megestrol Acetate; Melengestrol
Acetate; Melphalan;
Menogaril; Mercaptopurine; Methotrexate; Methotrexate Sodium; Metoprine;
Meturedepa;
Mitindomide; Mitocarcin; Mitocromin; Mitogillin; Mitomalcin; Mitomycin;
Mitosper; Mitotane;
Mitoxantrone Hydrochloride; Mycophenolic Acid; Nocodazole; Nogalamycin;
Ormaplatin; Oxisuran;
Paclitaxel; Pegaspargase; Peliomycin; Pentamustine; Peplomycin Sulfate;
Perfosfamide; Pipobroman;
Piposulfan; Piroxantrone Hydrochloride; Plicamycin; Plomestane; Porfimer
Sodium; Porfiromycin;
Prednimustine; Procarbazine Hydrochloride; Puromycin; Puromycin Hydrochloride;
Pyrazofurin;
Riboprine; Rogletimide; Safingol; Safingol Hydrochloride; Semustine;
Simtrazene; Sparfosate
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Sodium; Sparsomycinl, Spirogermanium Hydrochloride; Spiromustine; Spiroplatin;
Streptonigrin;
Streptozocin; Strontium Chloride Sr 89; Sulofenur; Talisomycin; Taxane;
Taxoid; Tecogalan Sodium;
Tegafur; Teloxantrone Hydrochloride; Temoporfm; Teniposide; Teroxirone;
Testolactone;
Thiamiprine; Thioguar~ine; Thiotepa; Tiazofurin; Tirapazamine; Topotecan
Hydrochloride;
Toremifene Citrate; Trestolone Acetate; Triciribine Phosphate; Trimetrexate;
Trimetrexate
Glucuronate; Triptorelin; Tubulozole Hydrochloride; Uracil Mustard; Uredepa;
Vapreotide;
Vertepor~n; Vinblastine Sulfate; Vincristine Sulfate; Vindesine; Vindesine
Sulfate; Vinepidine
Sulfate; Vinglycinate Sulfate; Vinleurosine Sulfate; Vinorelbine Tartrate;
Vinrosidine Sulfate;
Vinzolidine Sulfate; Vorozole; Zeniplatin; Zinostatin; Zorubicin
Hydrochloride.
Other anti-neoplastic compounds include: 20-epi-1,25 dihydroxyvitamin D3; 5-
ethynyluracil;
abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin;
ALL-TK antagonists; .
altretamine; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin;
amsacrine;
anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist
D; antagonist G;
antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen, prostatic
carcinoma; antiestrogen;
antineoplaston; antisense oligonucleotides; aphidicolin glycinate; apoptosis
gene modulators;
apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; arginine deaminase;
asulacrine; atamestane;
atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin;
azatyrosine; baccatin III
derivatives; balanol; batimastat; BCR/ABL antagonists; benzochlorins;
benzoylstaurosporine; beta
lactam derivatives; beta-alethine; betaclamycin B; betulinic acid; bFGF
inhibitor; bicalutamide;
bisantrene; bisaziridinylspermine; bisnafide; bistratene A; bizelesin;
breflate; bropirimine; budotitane;
buthionine sulfoximine; calcipotriol; calphostin C; camptothecin derivatives;
canarypox IL-2;
capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRest M3;
CARN 700; cartilage
derived inhibitor; carzelesin; casein kinase inhibitors (ICOS);
castanospermine; cecropin B;
cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin;
cladribine; clomifene
analogues; clotrimazole; collismycin A; collismycin B; combretastatin A4;
combretastatin analogue;
conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A
derivatives; curacin A;
cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate;
cytolytic factor; cytostatin;
dacliximab; decitabine; dehydrodidemnin B; deslorelin; dexifosfamide;
dexrazoxane; dexverapamil;
diaziquone; didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;
dihydrotaxol, 9-;
dioxamycin; Biphenyl spiromustine; docosanol; dolasetron; doxifluridine;
droloxifene; dronabinol;
duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab; eflornithine;
elemene; emitefur;
epirubicin; epristeride; estramustine analogue; estrogen agonists; estrogen
antagonists; etanidazole;
etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide;
filgrastim; finasteride;
flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicin
hydrochloride; forfenimex;
formestane; fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate;
galocitabine; ganirelix;
gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;
heregulin; hexamethylene
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bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene; idramantone;
ilmofosine; ilomastat;
imidazoacridones; imiquimod; immunostimulant peptides; insulin-like growth
factor-1 receptor
inhibitor; interferon agonists; interferons; interleukins; iobenguane;
iododoxorubicin; ipomeanol, 4-;
irinotecan; iroplact; irsogladine; isobengazole; isohomohalicondrin B;
itasetron; jasplakinolide;
kahalalide F; laxnellarin-N triacetate; lanreotide; leinamycin; lenograstim;
lentinan sulfate;
leptolstatin; letrozole; leukemia inhibiting factor; leukocyte alpha
interferon;
leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;
lineanpolyamine analogue;
lipophilic disaccharide peptide; lipophilic platinum compounds; lissoclinamide
7; lobaplatin;
lombricine; lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;
lurtotecan; lutetium
texaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A;
marimastat; masoprocol; maspin;
matrilysin inhibitors; matrix metalloproteinase inhibitors; menogaril;
merbarone; meterelin;
methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine;
mirimostim; mismatched
double stranded RNA; mitoguazone; mitolactol; mitomycin analogues; mitonafide;
mitotoxin
fibroblast growth factor-saporin; mitoxantrone; mofarotene; molgramostim;
monoclonal antibody,
human chorionic gonadotrophin; monophosphoryl lipid A+myobacterium cell wall
sk; mopidamol;
multiple drug resistance gene inhibitor; multiple tumor suppressor 1-based
therapy; mustard
anticancer agent; mycaperoxide B; mycobacterial cell wall extract;
myriaporone; N-acetyldinaline; N-
substituted benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin;
naphterpin;
nartograstim; nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase;
nilutamide; nisamycin;
nitric oxide modulators; nitroxide antioxidant; nitrullyn; 06-benzylguanine;
octreotide; okicenone;
oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral
cytolcine inducer; ormiaplatin;
osaterone; oxaliplatin; oxaunomycin; paclitaxel analogues; paclitaxel
derivatives; palauamine;
palmitoylrhizoxin; pamidronic acid; panaxytriol; panomifene; parabactin;
pazelliptine; pegaspargase;
peldesine; pentosan polysulfate sodium; pentostatin; pentrozole; perflubron;
perfosfamide; perillyl
alcohol; phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil;
pilocarpine hydrochloride;
pirarubicin; piritrexim; placetin A; placetin B; plasminogen activator
inhibitor; platinum complex;
platinum compounds; platinum-triamine complex; porfimer sodium; por~romycin;
propyl bis-
acridone; prostaglandin J2; proteasome inhibitors; protein A-based immune
modulator; protein kinase
C inhibitor; protein lcinase C inhibitors, microalgal; protein tyrosine
phosphatase inhibitors; purine
nucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;
pyridoxylated hemoglobin
polyoxyethylene conjugate; raf antagonists; raltitrexed; ramosetron; ras
farnesyl protein transferase
inhibitors; ras inhibitors; ras-GAP inhibitor; retelliptine demethylated;
rhenium Re 186 etidronate;
rhizoxin; ribozymes; RII retinamide; rogletimide; rohitukine; romurtide;
roquinimex; rubiginone B 1;
ruboxyl; safingol; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1
mimetics; semustine;
senescence derived inhibitor 1; sense oligonucleotides; signal transduction
inhibitors; signal
transduction modulators; single chain antigen binding protein; sizofiran;
sobuzoxane; sodium
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borocaptate; sodium phenylacetate; solverol; somatomedin binding protein;
sonermin; sparfosic acid;
spicamycin D; spiromustine; splenopentin; spongistatin l; squalamine; stem
cell inhibitor; stem-cell
division inhibitors; stipiamide; stromelysin inhibitors; sulfmosine;
superactive vasoactive intestinal
peptide antagonist; suradista; suramin; swainsonine; synthetic
glycosaminoglycans; tallimustine;
5 tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur;
tellurapyrylium;
telomerase inhibitors; temoporfin; temozolomide; teniposide;
tetrachlorodecaoxide; tetrazomine;
thaliblastine; thalidomide; thiocoraline; thrombopoietin; thrombopoietin
mimetic; thymalfasin;
thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin
ethyl etiopurpurin;
tirapazamine; titanocene dichloride; topotecan; topsentin; toremifene;
totipotent stem cell factor;
10 translation inhibitors; tretinoin; triacetyluridine; triciribine;
trimetrexate; triptorelin; tropisetron;
turosteride; tyrosine lcinase inhibitors; tyrphostins; UBC inhibitors;
ubenimex; urogenital sinus-
derived growth inhibitory factor; urolcinase receptor antagonists; vapreotide;
variolin B; vector
system, erythrocyte gene therapy; velaresol; veramine; verdins; verteporfm;
vinorelbine; vinxaltine;
vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; zinostatin stimalamer.
15 Anti-cancer supplementary potentiating agents include: tricyclic anti-
depressant drugs (e.g.,
imipramine, desipramine, amitryptyline, clomipramine, trimipramine, doxepin,
nortriptyline,
protriptyline, amoxapine and maprotiline); non-tricyclic anti-depressant drugs
(e.g., sertraline,
trazodone and citalopram); Caz+ antagonists (e.g., verapamil, nifedipine,
nitrendipine and caroverine);
calmodulin inhibitors (e.g., prenylamine, trifluoroperazine and clomipramine);
Amphotericin B;
20 Tripaxanol analogues (e.g., tamoxifen); antiarrhythmic drugs (e.g.,
quinidine); antihypertensive drugs
(e.g., reserpine); Thiol depleters (e.g., buthionine and sulfoximine) and
Multiple Drug Resistance
reducing agents such as Cremaphor EL.
The compounds of the invention also can be administered with cytolcines such
as granulocyte
colony stimulating factor or anti-sense oligonucleotides targeting survival
proteins such as, but not
25 limited to, Bcl-2, HIAPl, HIAP2, XIAP or surviving.
The conjugates of the invention also are useful, in general, for treating
mammalian cell
proliferative disorders other than cancer, including psoriasis, actinic
keratosis, etc.
The use of any of the compounds having the structure of formula I - III for
treatment andlor
prevention of neuological disorders, cancer, inflammation, or symptoms related
thereto is also
encompassed by the invention.
EXAMPLES
Examples of compounds according to formula I are provided in Table 2, based on
the
compound of formula IV (a subset of formula I), provided below. The compound
numbering used in
Table 2 to identify exemplary compounds is made reference to consistently
herein for all subsequent
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examples. For the structures in which XS is noted as meaning CH, this is
consistent with formula I as
XS-R5, wherein XS is C and RS is H.
H
A
A2 B2
R~ Xs Ra
/ N
\ ~ ~ ~~s~ / R~
N X
~4
R
IV
Table 2
Exemplary Structures of Formula VI, a Subset of Formula I
Compound Al/A2 B1/B2 Rl R4 R' Rg XS X9
1 O O H H H H CH CH
2 O O Me0 H H H CH CH
3 O O H H Me0 H CH CH
4 O O Me0 H Me0 H CH CH
5 O O Bn0 H H H CH CH
6 O O Bn0 CH2CHzOH H H CH CH
7 O O Bn0 CH2CHaOH Me0 H CH CH
8 O O Bn0 CH2CH20H H Me0 CH CH
9 O O Bn0 H H Me0 CH CH
10 O O H CHaCHaOH H H CH CH
11 O O Bn0 H Bn0 H CH CH
12 O O H H Bn0 H CH CH
13 O O Br H H H CH CH
14 O O I H H H CH CH
15 O O PhC---C H H H CH CH
16 O O PhCHzCH2 H H H CH CH
17 O O H H H H N CH
18 O O H C02tBu H H N CH
19 O O H COCH3 H H N CH
20 O O H CON(H)Ph H H N CH
21 O O Bn0 H H H N CH
22 O O H H H H CH N
23 O O H C02tBu H H CH N
Examples of compounds according to formula II, having a single bond between
carbon "a"
and XS are provided in Table 3, based on the compound of formula V (a subset
of Formula II),
provided below. The compound numbering used in Table 3 to identify exemplary
compounds is
made reference to consistently herein for all subsequent examples. XS is noted
as meaning CHZ
within this structure, which is consistent with formula II as XS-R5, wherein
XS is CH and RS is H.
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H
A~ N B~
A2 Bz
R~ Ra
IV
a 5~ ~ R~
N X
~4
R X5=CH2
V
Table 3
Exemplary Structures of Formula V, a Subset of Formula II
Compound Al/A2 B1B2 Rl R4 R' Rg
24 O O H H H H
25 sac-H/OH O H H H H
26 O sac-H/OH H H H H
27 HB O H H H H
28 O HB H H H H
29 O O H C02tBu H H
30 rac-HlOH O H C02tBu H H
31. O sac-H/OH H , COZtBu H H
32 HB O H COZtBu H H
33 O HB H C02tBu H H
34 O O H H MeO H
35 O O Bn0 H H H
36 O O Bn0 CH2CH20H H H
37 O O Bn0 CH2CHZOC(O)CHaOAc H H
38 O O Bn0 CHZCH20H Me0 H
39 O O Bn0 CHaCH20H H Me0
40 O O H CHZCH20H H H
41 O O H H Bn0 H
Examples of compounds according to formula II, having a double bond between
carbon "a"
and XS are provided in Table 4, based on the compound of formula VI ,(a subset
of Formula II),
provided below. The compound numbering used in Table 4 to identify exemplary
compounds is
made reference to consistently herein for all subsequent examples. XS is noted
as meaning CH within
this structure, which is consistent with formula II as XS-R5, wherein Xj is C
and RS is H.
R
R~
R'"
VI
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28
Table 4
Exemplary Structures of Formula VI, a subset of Formula II
Compound Al/AZ B1/BZ Rl R4 R~ XS
42 O O H H H CH
43 O rac-H/OH H H H CH
44 Yac-HlOH O H H H CH
~
45 O H/H H H H CH
46 H/H O H H H CH
47 O O Me0 H H CH
48 O O H H Me0 CH
49 O O Me0 H Me0 CH
50 O O Me0 COCH2N(H)C02tBu H CH
51 O O Me0 COCHZOC(O)CH3 H CH
52 O O Bn0 H H CH
53 O O H H H N
54 O O H COCH3 H N
55 O O H ~ COatBu H N
Examples of compounds according to formula III are provided in Table 5, based
on the
compound of formula VII (a subset of Formula III), provided below. The
compound numbering
used in Table 5 to identify exemplary compounds is made reference to
consistently herein for all
subsequent examples. Relative to formula III, the structures according to
formula VII have ring
members X', XZ and X3 represented by C (ring member X3 of formula III is shown
as "X" in formula
VII). RS is H for all exemplified compound in Table 5.
Examples of formula III are:
H
A2 ~B2
R~
i . , Y
~4
R
VII
Table 5
Exemplary Structures of Formula VII, a Subset of Formula III
Compound Al/Aa B1/B2 Rl RZ R3 R4 X Y
56 O O Br H H H C H
57 O ~ Br H H -COCH3 C H
O
58 O O Br H H -CONMea C H
59 ~ O O Br H H -COPh C H
60 O O Br H H Ts C H
61 O O Br H H -502(1-CIOH~) C H
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29
62 O O Br H H -S02(2-CIOH~) C H
63 O O Br H H dansyl C H
64 O O Bn0 H H H C H
65 O O Bn0 H H -COCH3 C H
66 O O Bn0 H H -COPh C H
67 O O BnO H H -CO(2,4-(Me0)2Ph)C H
68 O O Bn0 H H -CO(3,4-(Me0)2Ph)C H
.
69 O O Bn0 H H -COCH2N(H)Boc C H
~
70 O O Bn0 H H Ts C H
71 O O BnO H H -502(4-(N02)Ph) C H
72 O O Bn0 H H -S02(3-(N02)Ph) C H
73 O O Bn0 H H -CH2Ph C H
74 O O Bn0 H H -CH2(4-pyr) C H
75 O O Bn0 H H -CH2(3,5-(Me0)2Ph)C H
76 O O Bn0 H H 3-F(Ph)CH2- C H
77 O O Bn0 H H 4-F(Ph)CH2- C H
78 = O O Bn0 H H -CH2Phth C H
79 O O Bn0 H H -CH2(2-CIOH~) C H
80 O O Bn0 H H -CH2(C6H11) C H
81 O O Bn0 H H -CH2(CH2)6CH3 C H
82 O O Bn0 H H -CH2CH20H C H
83 O O Bn0 H H -CH2CH20Ac C H
84 O O Bn0 H H -CH2CH20- C H
CO(3,4-(Me0)2Ph)
85 O O Bn0 H H -CH2CH20C(O)NHPh C H
86 O O ~ MeO H H H C H
87 O O Me0 H H Ts C H
88 O O Me0 H H -S02(4-(NO2)Ph) C H
89 O O Me0 H H allyl C H
90 O O HO H H allyl C H
91 O O HO H H Ts C H
92 O O H H H H C H
93 O O H H H Ts C -H
94 O O H H H -502(4-(AcNH)Ph) C H
95 O O H H H -502(2-(N02)Ph) C H
96 O O H H H -S02(4-(N02)Ph) C H
97 O O H H H -S02Th C H
98 O O H H H -S02Bu C H
99 O O Cl H H H C H
100 O O H Cl H H C H
101 O O H H C1 H C H
102 O O F H H H C H
103 O O H F H H C H
104 O O N02 H H H C H
105 O S Bn0 H H H C H
106 O O H H H H C OMe
107 O O H H H H C OH
108 O O H H H Me C OH
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The compounds illustrated below as 109, 110 and 111 also fall within the
structure of formula
IH. Notably, for compound 109, the group representing RS is CHZ(CH3)ZCCOZMe.
H H
10 O"/O 111
G 1I
O_ 'NH
GENERAL PREPARATIVE METHODS
The compounds of the invention may be prepared by use of lrnown chemical
reactions and
procedures. Nevertheless, the following general preparative methods are
presented to aid the reader in
synthesizing compounds of formula I to III. Substitution patters have been
minimized' for
10 convenience except where clarification is required. However, all
combinations of substitution are
implicit in the methodologies outlined below.
The invention encompasses intermediates for manufacturing the compounds of
formula I to
III, as described herein. Mixtures including isomeric mixtures also may result
depending upon the
symmetry of the starting molecule. Such mixtures are within the scope of the
invention.
15 To prepare the full range of compounds of the invention, only the chemistry
described below,
together with chemistry lrnown to those of skilled in the art is required. In
particular, modifications of
the core structures can be accomplished using routine chemistry such as
described herein.
Method A: Synthesis of 3-(indol-3-yl)-4-(1N indolyl)-1H pyrrole-2,5-diones
20 Indole, intermediate Al, is treated with oxalyl chloride, in a solvent such
as THF or diethyl
ether, to provide the 3-indolylglyoxalyl chloride hydrochloride salt, which is
further treated with
NaOMe in MeOH, to yield the methyl 3-indolylglyoxylate, intermediate Bl. A
second indole, Al, is
converted to the acetamide intermediate Cl, by stirring A1 with NaH in DMF,
followed by the
addition of iodoacetamide. Intermediates B1 and Cl are suspended in THF and
treated with 3 to 4
25 equivalents of a base such as KOtBu, to provide compound 1, which falls
under the general class of 3-
(indol-3-yl)-4-(1N indolyl)-1H pyrrole-2,5-diones.
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_ O
1 ) (COCI)2 \ / OMe
N 2) NaOMe/MeOH N J O
' KOtBu
H
A1 B1 ~ THF
\ 1 ) NaH, DMF
N 2) ICH2CONH2 N
CH2CONH2
A1 C1
The synthesis according to Method A can be generally described by the above
reactions. The
invention relates to the above-noted method for preparation of 3-(indol-3-yl)-
4-(1N indolyl)-1H
pyrrole-2,5-dione compounds, said method comprising the following steps: (a)
reacting indole with
oxalyl choride in a solvent to form a hydrochloride salt; (b) treating said
hydrochloride salt with
NaOMe in alcohol to form a methyl 3-indolglyoxylate; (c) reacting indole with
a strong base in a
polar solvent; (d) reacting the product of step (c) with haloacetamide to form
an acetamide
intermediate; and (e) treating the products of steps (b) and (d) with excess
base to form a 3-(indol-3-
yl)-4-(1N indolyl)-1H pyrrole-2,5-dione. It is implicit in this method that
the reactants may be
substituted in such a way that the resulting compound is a 3-(indol-3-yl)-4-
(1N indolyl)-1H pyrrole-
2,5-dione, which may for example fall within the structure of formula I.
Further, the product of step
(e) may be further reacted to add such substituents as those noted within the
structure of formula I.
Method B: Synthesis of 3-(indol-3-yl)-4-(1N indolyl)-1H pyrrole-2,5-diones
Intermediate A1 is treated with oxalyl chloride in a solvent such as diethyl
ether or THF. The
resulting 3-indolylglyoxalyl chloride hydrochloride salt is treated with
aqueous ammonium carbonate
to provide the acetamide intermediate Dl. A second appropriately substituted
indole, Al, is treated
with a KOtBu, followed by ethyl bromoacetate to yield intermediate El (in
situ). To this solution of
El is added 1 equiv of solid Dl. The resulting suspension is treated with 5
equiv of KOtBu and
stirried over night. After quenching with conc HZS04, aqueous worlcup and
purification by silica gel
chromatography, compound 1 is isolated in low yield.
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_ o
\ 1) (COCI)2 \ / NH2
2) aqu NH4C03 ~ ~ H
O N O
A1 D1 KOtBu
THF i
1 ) base, THF
/ \ ~ / \
2 BrCH CO Et N
N ) z 2 i
CH2COZEt 1
H
A1 E1 in situ
The synthesis according to Method B can be generally described by the above
reactions. The
invention relates to the above-noted method for preparation of 3-(indol-3-yl)-
4-(1N indolyl)-1H
pyrrole-2,5-dione compounds, said method comprising the following steps: (a)
reacting indole with
oxalyl choride in a solvent to form a hydrochloride salt; (b) treating said
hydrochloride salt with
aqueous ammonia to form an acetamide intermediate; (c) reacting indole with a
base; (d) reacting the
product of step (c) with haloacetate; and (e) adding an equivalent of the
product of step (b) to the
product of step (d) and treating with excess base to form a 3-(indol-3-yl)-4-
(1N indolyl)-1H pyrrole-
2,5-dione. It is implicit in this method that the reactants may be substituted
in such a way that the
resulting compound is a 3-(indol-3-yl)-4-(1N indolyl)-1H pyrrole-2,5-dione,
which may for example
fall within the structure of formula I. Further, the product of step (e) may
be further reacted to add
such substituents as those noted within the structure of formula I.
Method C: Cyclization of 3-(indol-3-yl)-4-(1N indolyl)-1H pyrrole-2,5-diones
Compound 1 (Rl=H) was suspended in a solvent such as methlyene chloride and
treated with
1.2 equiv of TMSOTf. After stirring 0 to 24 hours the solvent is removed to
provide, after silica gel
chromatography, compound 24 (R'=H). When Rl is MeO, compound 2, cyclization is
followed by
autooxidation to provide compound 47.
When R' is BnO, compound 5, BF30Et2 is added to a THF solution of 5, and the
reaction is
not stirred for more than 6 hours, to provide compound 35.
H
i
O N O
R3 ~ - N ~ Lewis Acid R~
~ I N ~ 1
H
H
1; R3=H 24; R3=H
2; R3=Me0 35; R3=Bn0
5; R3=Bn0
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The synthesis according to Method C can be generally described by the above
reactions. The
invention relates to the above-noted method for cyclization of a 3-(indol-3-
yl)-4-(1N indolyl)-l~I
pyrrole-2,5-dione compound, said method comprising the step of reacting a 3-
(indol-3-yl)-4-(1N
indolyl)-1H pyrrole-2,5-dione with a Lewis acid to form a pyrrolo-a-hydro-[3-
carboline. The product
of this method may fall within the structure of formula II. Further, the
product of the method may be
further reacted to add such substituents as those noted within the structure
of formula II.
Method D: DDQ Oxidation of Dihydro-pyrrolo-(3-carbolines
Compound 24 is dissolved in a solvent such as 1,4-dioxane and treated with a
dioxane
solution of an oxidizing agent, in this case 2,3-dichloro-5,6-dicyano-1,4-
benzoquinone (DDQ) (2.2
equiv). Filtration through celite and purification by silica gel
chromatography provides compound 42.
H H
O N O O N O
w DDQ
~N / . ~N \ ~ /
H
H
24 42
The synthesis according to Method D can be generally described by the above
reaction. The
invention relates to the above-noted method for oxidation of a dihydro-pyrrolo-
~3-carbolines, said
method comprising the step of reacting a pyrrolo-a-hydro-(3-carboline of
formula 1I, having a single
bond at carbon (a) with an oxidizing agent. The product of this method may
fall within the structure
of formula II, having a double bond at carbon (a). Further, the product of the
method may be further
reacted to add such substituents as those noted within the structure of
formula II.
Method E: Preparation of N Ethylhydroxy Derivatives
Intermediate A1 was converted to its N indolyl acetic acid derivative,
intermediate Fl, using
phase transfer catalysis. Subsequent LAH reduction to the corresponding acid,
intermediate Gl, and
acylation provided intermediate Hl. Conversion of Hl to its methyl glyoxylate,
intermediate Bll,
followed as described above in Method A.
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Et0 CCH Br
z 2 \ ~ \ LiAIHa \ ~ \
I -
\ ~ cat. n-Bu4NHS04 N N
50%nNaOH ~CO H
z
A1 OH
F1 G1
_ O
Ac20 \ ~ \ ~ ) (COCI)z ~ ~ OMe
N ~ O
2) NaOMe NJ
OAc
H1 HO B11
Coupling of Cl with Bll, as described in method A, yielded compound 10.
Cyclization of 10
using BF30Et2 at-7S °C, provided the desired di-hydro-(3-carboline;
compound 40.
I~OtBu, THF BF30Et2
B11 --
THF, -78 °C
N C1
'CONH2
Method F: Carbonyl Reduction and Sulfonylation Reactions
Compounds 24 and 29 are readily reduced to a mixture of alcohols. Compound 24
(R4=H)
provides a 1:1 mixture of 25 and 26, while compound 29 (R= COZtBu) provides a
3:1 mixture of 30
and 31. These compounds were separable by silica gel chromatography. ,
O N O
NaBH4
w
N ~ / MeOH
N
R4 R R
24; R4=H . 25; R4=H 26; R4=H
29; R4=CO~tBu 30; R4=COZtBu 31; R4=C02tBu
a H H
PhSH PhS N O O N SPh
cat TsOH H20
-v + w
25 + 26 enzene ~ N ~ i N
reflux
N N
H H
27 28
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Treatment of either the mixture of compounds 25 and 26, or each
independently,. with PhSH
and catalytic TsOH yields compound 27 and 28, respectively. This chemistry is
also applicable to the
fully oxidized (3-carbolines such as compound 42.
5 Treatment of either the mixture of 30 and 31, or each independently, with 2
equiv of PhSeH
and a catalytic amount of TsOH yields compound 32 and 33, respectively.
H
O N
2 PhSeH
cat TsOH H20
30 + 31 CH2CI2/MeOH \ I \ N
N
Boc Boc
32 33
Method G: Palladium Catalyzed Indole Functionalization and Glyoxylate
Formation
10 Intermediate A7 was prepared by the treatment of 5-iodoindole, A6, with
(Boc)z0. Palladium
catalyzed coupling of A7 with phenylacetylene proceeded in good yields to
provide intermediate A8,
which was readily deprotected by photolysis in a solvent such as acetonitrile,
with a 250 W light bulb,
to provide intermediate A9. Conversion of A9 was to the methyl glyoxylate
intermediate B9, followed
as described in Method A.
Ph
I ~ \ 5% (PPh)2PdCl2
N + Ph - EtZN'Pr, THF I / N
Boc reflux 24 hrs Boc
A6 A7
Ph Ph O OMe
\ 1 ) (COCI)2 \\
\ \ ~O
ACN, 16 hrs I / N 2) NaOMe
N
H H
A8 gg
Intermediate A8 was reduced using tosylhydrazine and sodium acetate to yield
intermediates
A9, which was then converted to the methyl glyoxylate intermediate B9, as
described in Method A.
Ph Ph
Ph \ \ \ H2 Pd/C ~ \ 1 ) (COC12) O OMe
N~ MeOH , ~ N~ 2) NaOMe l \ ~ O
H H ' N
A8 A9 H B9
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The synthesis according to Method G can be generally described by the above
reactions. The
invention relates to the above-noted method for preparation of a
functionalized methyl glyoxolate
indole, the method comprising the following steps: (a) reacting N Boc-
iodoindole with an acetylene in
the presence of a palladium catalyst under coupling,conditions; (b)
deprotecting the product of step (a)
by photolysis in solvent; (c) reacting the product of step (b) with oxalyl
choride to form a
hydrochloride salt; and (d) forming a methyl 3-(acetyleno)indolglyoxylate by
treating said
hydrochloride salt with NaOMe.
The product of this reaction may be used to form functionalized compounds
according to the
invention, for example as an intermediate in synthetic routes described in
Methods A and B, above.
Method H: Acylation of pyrrolo-(3-Carboline Derivatives with a,-Amino Acids
A THF solution of compound 47, Boc-Gly-OH, DIC, Et3N, DMAP (cat) is refluxed
for 24
hours. This reaction mixture is subjected to standard aqueous worle up and
purification by silica gel
chromatography, to yield the acylated product, compound 50.
H
O N O
Boc-Gly-OH i0 , N
DIC, Et3N, DMAP ~ I N~ ~ l
THF, reflux
H 60% O~ O
47 H,N
50 \\O
Method I: Acylation of (3-Carboline Derivatives with Acid Chlorides
A THF solution of compound 47, acetoxyacetyl chloride, Et3N, DMAP (cat) is
stirred at room
temperature for 24 hours. This reaction mixture is subjected to standard
aqueous work up and
purification by silica gel chromatography, to yield the acylated product,
compound 51.
H
CICOCH20Ac
Et3N, DMAP
THF, reflux
47
Method J: Synthesis of 3-(indol-3-yl)-4-(1N benzyimidazolyl)-1H pyrrole-2,5-
diones
Benzimidazole (2 equiv) was stirred overnight, in a solvent such as THF, with
iodoacetamide
(1 equiv) to yield acetamide Il. Acetamide Il, glyoxylate Bl, KZC03, and CETAB
are susbended in
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benzene and refluxed for 5 days, while removing water with a Dean Stark trap.
Aqueous work up and
purification by silica gel chromatography yields two products, compound 17 and
its methyl ester Jl.
N i O THF
~NHZ over night N
N
~CONHz
11
O -- . O N~ OMe
OMe ~~ KzCOa
N CETAB ~ + ~ N w
O ' benzene ~ ~ ~ y
H CONHZ reflux 5 d N N
H
11 17 ' J1
Alternatively, compounds Bl and Il may be combined in THF and treated with a
THF
solution of KOtBu (3 equiv). The reaction is stirred for 1 hour before aqueous
workup and
purification by silica gel chromatography provides 17 and variable quantities
of 106.
O
OMe + ~ ~ ~ 31<OtBu
NJ O N THF, 1 h .
~CONH2
H
B1 11 17 106
The synthesis according to Method J can be generally described by the above
reactions. The
invention relates to the above-noted method for preparation of 3-(indol-3-yl)-
4-(1N benzyimidazolyl)-
1H pyrrole-2,5-diones compounds, said method comprising the following steps:
(a) reacting indole
with oxalyl choride in a solvent to form a hydrochloride salt; (b) treating
said hydrochloride salt with
NaOMe in alcohol to form a methyl 3-indolglyoxylate; (c) reacting
benzoimidazole with a strong base
in a polar solvent; (d) reacting the product of step (c) with haloacetamide to
form an acetamide
intermediate; and (e) treating the products of steps (b) and (d) with excess
base to form a 3-(indol-3-
yl)-4-(1N benzyimidazolyl)-1H pyrrole-2,5-dione.
Method K: Acylation and Cyclization of 3-(indol-3-yl)-4-(1N benzyimidazolyl)-
1H pyrrole-2,5-diones
Compound 17 ~is readily acylated with acid chlorides, anhydrides, and
isocyanates, to yield
compounds 18, 19, and 20, respectively. Photolysis of compounds 18 and 19 (300
watt bulb, 48 hrs,
acetonitrile) provides compounds 54 and 55, respectively. Photolysis of
compound 20, as above,
yields the cyclized and de-carbonylated compound 53.
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H
O N O
(RCO)~O ~ i N
2COCI CH3CN ~ N N
R~O
18; R=CH3 54; R=CH3
19; R=OC(CH3)3 55; R=OC(CH3)s
17
PhNCO
THF
CH3CN
53
The synthesis according to Method K comprises the following methodological
steps: (a) reacting 3-
(indol-3-yl)-4-(1N benzyimidazolyl)-1H pyrrole-2,5-dione with an acylating
agent selected from the
group consisting of anhydride, acid chloride and isocyanate to provide 3-(N
acylindol-3-yl)-4-(1N
benzyimidazolyl)-1H pyrrole-2,5-dione; and (b) photolysis of the product of
step (a) in a solvent to
form cyclization of 3-(indol-3-yl)-4-(1N benzyimidazolyl)-1H pyrrole-2,5-
diones.
Method L: Synthesis of 3-(1-methylindol-3-yl)-4-(benzyimidazol-1-yl)-1-
acetylpyrrole-2,5-
dione and 3-(1-methylindol-3-yl)-4-(imidazol-1-yl)-1-acetylpyrrole-2,5-dione
To a THF solution of compound 108 and a base such as triethylamine (4 equiv),
is added 3
equiv of acetyl chloride.
Ac Ac
N O 1 ) 3 AcCI, 4 Et3N O N O O N O
THF
w w
or
OH 2) imidazole or i I N i N
\ \ /
\ benzimidazole
m
108 . ~ .. .; ..
Me Me Me
K1 L1
This solution is stirred for 2 hours before either imidazole or benimidazole
is added. After
stirring for 2 to 24 hours intermediates Kl or Ll, respectively, was obtained
after purification by
recrystallization or silica gel chromatography.
Method M: One Pot preparation of 3-(indol-3-yl)-1H pyrrole-2,5-diones
Indole A1 (1.0 equiv) is dissolved in a solvent such as diethyl ether or THF
and treated with
oxalyl chloride ( 1.1 equiv). After stirring at room temperature for 1 to 24
hours the volatiles are
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removed under reduced pressure. Acetamide (3 equiv) is added to the resulting
3-indolylglyoxalyl
chloride hydrochloride salt, and this mixture is taken up in dry THF. After
stiring at room
temperature for 2 to 3 hours a 1.0 M THF solution of I~OtBu (5 equiv) is
added. Ths resulting purple
solution is stirred for 3 to 24 hours before the reaction is quenched with
cone HzSOø (30 minutes at
room temperature), followed by aqueous extraction and purification by
recrystallization or silica gel
chromatography to yield compound 92.
1 ) (COCI)2, Et20
-' 2) acetamide, THF
3) 3 KOtBu, THF
4) conc H2S04
A1 92
The one pot synthesis according to Method M can be generally described by the
above
reaction. The invention relates to the above-noted method for preparation of a
3-(indol-3-yl)-1H
pyrrole-2,5-dione compound, said method comprising the following steps: (a)
dissolving indole in a
solvent and treating with oxalyl choride to form a hydrochloride salt; (b)
treating said hydrochloride
salt with acetamide in solvent; (c) reacting the product of step (b) with
excess strong base in THF; and
(d) reacting the product of step (c) with strong acid to form 3-(indol-3-yl)-
1H pyrrole-2,5-dione.
Method N: . Acylation and Sulfonylation of 3-(1H indol-3-yl)-1H pyrrole-2,5-
diones
The disclosed 3-(1H indol-3-yl)-1H pyrrole-2,5-diones were readily acylated
with a number
of anhydrides, mixed anhydrides, and acid chlorides. Acylations using acetic
anhydride or acetyl
chloride were complete within 1 hour, however, aryl acid chlorides and mixed
anyhydrides required
refluxing over night in THF, with triethylamine and catalytic DMAP.
H
O N O
Ac~O gn0 PhCOCI
i
N, DMAP ~ ~ ~ Et3N, DMAP
THF N THF
H
65 s4
H
O N O
O
Boc(H)N~ Bn0
64 X ~ ~ ~ 69
Et3N, DMAP, THF N
X=OSuc, OPf,
OPiv
or DIC, cat DMA \
P
when X=OH N(H)Boc
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Similarly, Boc-Gly-OH couples with 64 in the presence of DIC and cat. DMAP in
refluxing
THF. Sulfonylation with various sulfonyl chlorides, triethylamine, and up to 2
equiv of DMAP, in
THF, required refuxing for 48 hours and provided the desired sulfonamides in
low to moderate yields.
H
H O N O
O N O
TsCI Bn0
Bn0
Et3N, DMAP \ I N
\ N THF, reflux , _O
, S~
H ~ ~O
64 ~ a 70
5 Method O: Functionalization of Compound 82
Indole A3 was converted to intermediate H3 via the three step process
described in Method E.
Intermediate H3 was converted to compound 82 using Method M.
B
Bn0 1) BrCH2C02Et, n-Bu4NHS04
benzene, 50% NaOH H3
2) LiAIH4, THF
3) Ac20, Et3N, THF
A3 95% for 3 steps H
I
O N O
1 ) (COCI)2, Et20
2) acetamide, THF Bn0
82
3) 3 KOtBu, THF \ N
4) cons HCI
to 65% OH
Compound 82 was acylated in a similar manner as that described in Method N, to
provide
10 compounds 83 and 84. Treatment of 82 with phenylisocyanate provides
compound 85.
H H
Bn RCOCI Bn PhNCO B
_t3N, THF _t3N, THF
reflux
~ OH
R
~~; K=~~s 82 85
84; R=(3,4-(Me0)2Ph)
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PREPARATIVE METHODS FOR INTERMEDIATE COMPOUNDS
Preparation methods of intermediate compounds for synthesis of compounds of
formula I to
III are within the scope of the invention, as are the intermediate compounds
themselves. Various
commercially available indoles were used and are labled as follows; indole,
intermediate Al; 5-
methoxyindole, intermediate A2; ~5-benzyloxyindole, intermediate A3; 5-
bromoindole, intermediate
A4; 5-iodoindole, intermediate A5. Exemplary Preparative methods are provided
below
Intermediate A6: N Boc-5-iodoindole
Di-teYt-butyl dicarbonate (2.2 mL; 9.05 mmol) was added to a solution of 5-
iodoindole (2.00 g, 8.23
mmol) in THF (50 mL) followed by a catalytic amount of DMAP and left to stir
at room temperature
for 10 minutes. The reaction was diluted with ethyl acetate (200 mL), the
organic solution washed
with saturated ammonium chloride solution (100 mL), water (100 mL) and brine
(100 mL). The
organic phase was dried over anhydrous magnesium sulfate and concentrated in
vacuo to yield
Intermediate A6 as an off white solid, which was used without further
purification. 1H NMR
(200MHz, CDCl3) 8 7.91 (d, J=7.1 Hz, 1H), 7.88 (s, 1H), 7.59 (d, J=7.lHz, 1H),
7.52 (d, 2.SHz, 1H),
6.45 (d, J=2.SHz, 1H), 1.65 (s, 3H).
Intermediate A7: N Boc-5-(2-phenethyl)indole
Pd(PPh3)ZCl2 (288 mg, 0.411 mmol) and CuI (157 mg, 0.822 mmol) were suspended
in THF (50 mL)
and the solution purged with NZ for 10 minutes. Intermediate A6 (2.82 g, 8.22
mmol) and triethyl
amine (50 mL) were added and the solution purged with NZ for a further 10
minutes. Phenyl
acetylene (903 p,1; 8.22 mmol) was added and the reaction left to stir at room
temperature 'for 2 hours.
The solvent was removed in vacuo and the material purified by silica gel
chromatography, eluting
with 2:1 hexanes/ethyl acetate, to provide intermediate A7 as an off white
solid (2.05 g, 78.5%). m.p.
106-107 °C. 1H NMR (500 MHz, DMSO-d6) 1.61 (9H, s), 6.72 (1H, d, J3.7),
7.42 (3H, m), 7.48 (1H,
dd, J 8.5, 1.2), 7.55 (1H, dd, J 7.7, 2.1), 7.71 (1H, d, J 3.7), 7.83 (1H, s),
8.07 (1H, d, J 8.5).
Intermediate A8: 1H 5-(2-phenethyl)indole
A solution of intermediate A7 (1.20 g, 3.78 mmol) in acetonitrile (50 mL) was
placed directly above a
150W light bulb and left for 16 hours. The solvent was removed in vacuo and
the residue purified by
silica gel chromatography, eluting with 3:1 hexanes/ethyl acetate, to yield
intermediate A8 as a yellow
solid (700 mg, 85 %).
m.p. 133-135°C. 'H NMR (500 MHz, DMSO-d6) 8 6.47 (1H, m), 7.26 (1H, dd,
J 8.4, 1.5), 7.39 (4H,
m), 7.44 (1H, d, J 8.4), 7.53 (2H, dd, J 8.4, 1.5), 7.79 (1H, d, J 0.6), 11.32
(1H, s).
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Intermediate A9: 5-(2-phenethy)lindole
A solution of intermediate A8 (2.05 g, 6.5 mmol) andp-toluenesulfonohydrazide
(12.1 g, 6.5 mmol)
in 1,2-dimethoxyethane (80 mL) was heated to reflux. A solution of sodium
acetate (8.9 g, 0.1 mol)
in water (80 mL) was added dropwise over a period of 4 hours maintaining the
reaction at reflux. The
reaction was left to reflux for a further 16 hours, cooled to room temperature
and poured onto water
(500 mL). The solution was extracted with dichloromethane (3 x 300 mL), the
organic extracts
combined, washed with water (300 mL), dried over anhydrous magnesium sulfate
and concentrated in
vacuo. Purification by silica gel chromatography, eluting with 3:1
hexanes/ethyl acetate, yielded
intermediate A9 as an off white solid. 1H NMR (200MHz, CDCl3) 8 8.03 (br s,
1H), 7.50 (s, 1H),
7.40-7.15 (m, 7H), 7.06 (dd, J=1.2, S.OHz, 1H), 6.52 (s, 1H), 3.02 (s, 4H).
Intermediate B1: Methyl 3-indoleglyoxylate
Oxalyl chloride (1.50 mL; 17.2 mmol) was added dropwise to an ice cold
solution of indole, A1, (2.00
g; 17.1 mmol) in anhydrous diethyl ether (20 mL). The resulting solution was
allowed to stir on ice
for 1 hour after which time a yellow slurry had formed. A freshly prepared
solution of sodium
methoxide in methanol (780 mg Na metal in 20 mL methanol; 34.1 mmol) was
added, the reaction
allowed to warm to room temperature and stirred for 1 hour. The reaction was
quenched with the
addition of water (30 mL) and the resulting orange solid isolated by ~tration,
washed with diethyl
ether and recrystallised from methanol. Yield 2.94 g; 85 %. mp 226-228
°C. 1H NMR (500 MHz,
DMSO-d6) 12.40 (br s, 1H), 8.17 (dd, J=1.5, 7.6Hz, 1H), 7.55 (dd, J=1.5,
7.6Hz, 1H), 7.29 (dt, J=1.5,
7.6Hz, 1H), 7.26 (dt, J=1.5, 7.6Hz, 1H), 3.89 (s, 3H).
Intermediate B2: Methyl 5-methoxy-3-indoleglyoxylate
5-methoxy-indole (2.00g, 13.6mmol) was dissolved in diethyl ether and stirred
at 0°C for 15 min.
Oxalyl chloride (1.30m1) was added via syringe and the reaction stirred at RT
for 3h. During this time
a precipitate formed. A solution of sodium methoxide in methanol (27.2 mmol)
was then added using
a syringe at -78 °C. The reaction was then brought to RT and stirred
for 3h, until it was quenched with
water (15m1). A solid yellow precipitate formed, and was filtered off, before
being dried in vacuo to
yield intermediate B2 as a yellow solid (2.10 g, 66%). m.p. 251.6-252.7
°C. 1H NMR (200 MHz,
DMSO-d6) 8 12.60 (br s, 1H), 8.35 (d, J=3.OHz, 1H), 7.65 (d, J=2.4Hz, 1H),
7.46 (d, J=8.8Hz, 1H),
6.91 (dd, J=2.6, 8.9Hz, 1H), 3.88 (s, 1H), 3.79 (s, 1H).
Intermediate B3: Methyl 5-benzyloxy-3-indoleglyoxylate ,
Intermediate B3 was prepared according to the method described for
Intermediate Al, using 5-
benzyloxy-indole, A3, (1.91 g; 8.55 mmol), oxalyl chloride (750 ~L; 8.6
rnmol), 5-benzyloxy-indole,
A3, (1.91 g; 8.55 mmol), anhydrous diethyl ether (10 mL), and a freshly
prepared solution of sodium
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methoxide in methanol (390 mg Na metal in 10 mL methanol; 17.1 mmol), to yield
intermediate B3
as a yellow solid (2.07 g, 78 %). mp 254-255 °C.
Intermediate B4: Methyl 5-bromo-3-indoleglyoxylate
Intermediate B3 was prepared according to the method described for
Intermediate Al, using 5-
benzyloxy-indole, A3, (1.91 g; 8.55 mmol), oxalyl chloride (750 ~,L; 8.6
mmol), 5-benzyloxy-indole,
A3, (1.91 g; 8.55 mmol), anhydrous diethyl ether (10 mL), and a freshly
prepared solution of sodium
methoxide in methanol (390 mg Na metal in 10 mL methanol; 17.1 mmol), to yield
intermediate B3
as a yellow solid (2.07 g, 78 %). 'H NMR (200MHz, DMSO-d6) 8 12.66 (br s),
8.51 (dd, J=0.4,
3.3Hz, 1H), 8.28 (s, 1H), 7.50 (t, J=7.6Hz, 1H), 7.45 (t, J=7.6Hz, 1H), 3.89
(t, 3H).
Intermediate B5: Methyl 5-iodo-3-indoleglyoxylate
Intermediate B5 was prepared according to the method described for
Intermediate B1, using 5-
iodoindole, A5, (1.0 g, 4.11 mmol), oxalyl chloride (361 ~L; 4.14 mmol),
anhydrous diethyl ether (5
mL), and a freshly prepared solution of sodium methoxide in methanol 190 mg Na
metal in 5 mL
methanol; 8.22 mmol), to yield intermediate B5 as a yellow solid (501 mg, 37
%). m.p. 280-281°C. 1H
NMR (500 MHz, DMSO-d6) ~ 3.88 (3H, s), 7.40 (1H, d, J 8.6), 7.57 (1H, dd, J
8.5, 1.7), 8.44 (1H, d,
J3.3), 8.49 (1H, d, J 1.7), 12.6 (1H, br).
Intermediate B8: Methyl 5-(2-phenethyl)-3-indole glyoxylate
Intermediate B8 was prepared according to the method described for
Intermediate B1, intermediate
A8 (400 mg, 1.84 mmol), oxalyl chloride (321 ~L; 3.68 mmol), anhydrous diethyl
ether (10 mL), and
a freshly prepared solution of sodium methoxide in methanol (85 mg Na metal in
5 mL methanol;
3.68 mmol), to yield intermediate B5 as a yellow solid (301 mg, 54 %). m.p.
275-279 °C. HRMS
Cal'd for C19HI3NO3 303.0895. Found 303.0899.
Intermediate B9: Methyl 5-(2-phenethyl)-3-indoleglyoxylate
Intermediate B9 was prepared according to the method described for
Intermediate B1, intermediate
A9 (300 mg, 1.40 mmol), oxalyl chloride (147 ~,L; 1.68 mmol), anhydrous
diethyl ether (10 mL), and
a freshly prepared solution of sodium methoxide in methanol (64 mg Na metal in
5 mL methanol,
2.80 mmol), to yield intermediate B5 as a yellow solid (327 mg, 76 %). m.p.
204-207 °C. HRMS
Cal'd for C19H1~N03 307.1208. Found 307.1206.
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Intermediate B 10: Methyl 1-(2-hydroxyethyl)-5-benzyloxy-3-indoleglyoxylate
Intermediate Al l (5.64g, 18.37 mmol) was dissolved in dry ether (250 mL).
Oxalyl chloride (1.77
mL) was added via syringe and the reaction allowed to stir for 3 hours. In a
seperate flask Na metal
(1.39 g, 55.11 mmol) is dissolved slowly into omnisolv methanol (50 mL). After
3 hours the sodium
methoxide solution is slowly added to the original reaction at 0°C. The
reaction is then allowed to stir
over night at room temperature. After 24 hours water (50 mL) is added to
quench the reaction. A large
amount of solid precipitates out. The solid is filtered off and washed with
water (20 mL) and
methanol (3 x 20 mL). This afforded intermediate B11 as a yellow solid (3.35
g, 52%). m.p. 148.2-
149.5 °C. 1H NMR (200MHz, DMSO-d6) 8 10.52 (br s, 1H), 7.98 (s, 1H),
7.43 (m, 6H), 7.11 (d, 9.0
Hz, 2H), 6.96, (m, 2H), 6.72 (m, 2H), 5.30 (dd, 5.2 Hz, 4.5 Hz, 1H), 5.10 (d,
3.7 Hz, 2H), 4.91, (t, 5.2
Hz, 1H), 4.26 (m, 2H), 3.69 (m, 2H).
Intermediate B 1 ~1: Methyl 1-(2-hydroxyethyl)-3-indoleglyoxylate
Oxalyl chloride (663 ~,1; 7.6 mmol) was added dropwise to an ice-cold solution
of intermediate Al l
(780 mg; 3. 8mmo1) in diethyl ether (5 mL). The solution was allowed to warm
to room,temperature
and stirred for 2 hours. The reaction was cooled on ice and a freshly prepared
solution of sodium
methoxide in methanol (262 mg Na metal in 8rnL methanol) added. The reaction
was allowed to
warm to room temperature and stirred for 3 hours. The reaction was quenched by
the addition of
water (10 mL) and the resulting pale brown solid isolated by filtration,
washed with water, diethyl
ether and dried irt vacuo to furnish intermediate B8 (550 mg, 59%). m.p. 131-
132 °C. 1H NMR (500
MHz, DMSO-d6) 8 3.76 (2H, dt, J 5.3, 5.3), 3.89 (3H, s), 4.36 (2H, t, J 5.3),
4.98 (1H, t, J 5.3), 7.30
(1H, ddd, J7.1, 7.1, 1.0), 7.33 (1H, ddd, J7.2, 7.2, 1.4), 7.66 (1H, d, J7.6),
8.18 (1H, d, J7.7), 8.46
(1H, s).
Intermediate C1: 2-(N indolyl) acetamide
A solution of indole (10 g; 0.085 mol) in DMF (50 mL) was added dropwise to an
ice cooled
suspension of NaH (supplied as 60% dispersion in mineral oil) (4.1 g; 0.10
mol) in DMF (100 mL).
The reaction was allowed to warm to room temperature and stirred for 4 hours
until all NaH was
consumed. A solution of iodoacetamide (18.5 g; 0.1 mol) in DMF (50 mL) was
added and the
reaction left to stir at room temperature overnight. The DMF was removed irt
vacuo and the residue
dissolved in ethyl acetate (1 L). The organic solution was washed with water
until the washings were
clear (3 x 500 mL), brine (500 mL), dried over anhydrous magnesium sulfate and
the solvent removed
ita vacuo. Recrystallisation from ethyl acetate provided intermediate C1 as an
off white solid (9.20 g,
62 %). m.p. 177-179 °C. 1H NMR (500 MHz, DMSO-d6) 8 4.79 (2H, s), 6.44
(1H, dd, J 3.1, 0.7), 7.03
(1H, ddd, J7.8, 7.8, 0.9), 7.13 (1H, ddd, J8.2, 8.2, 1.0), 7.24 (1H, br), 7.31
(1H, d, J3.1), 7.35 (1H,
dd, J 8.2, 0.6), 7.50 (1H, br), 7.55 (1H, d, J7.8).
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Intermediate C2: 1H 5-methoxyindol-3-yl acetamide
5-methoxyindole (2.00 g, 13.6 mmol) was dissolved in DMF (75 mL). Sodium
hydride (652 mg,
16.31 mmol) was added quickly and the reaction was stirred for 4 hours.
Iodoacetamide (2.51g, 13.6
5 mmol) was then added and the reaction allowed to stir over night at room
temperature. The DMF is
then removed on the rotary evaporator and the resulting solid worked up in
ethyl acetate and water.
The recovered solid is then purified through 2 recrystallizations in ethyl
acatete. This yields the white
solid intermediate C2 (1.95 g, 71%). m.p. 201.1-202.8 °C.'H NMR
(200MHz, DMSO-db) 8 7.55(br s,
1H), 7.12 (m, 3H), 7.04 (s, 1H), 6.75 (d, 7.25 Hz, 1H), 6.33, (d, 3.4 Hz,lH),
6.74(s, 2H), 6.73(s, 3H).
Intermediate C3: 1H 5-benzyloxyindol-3-yl acetamide
A solution of 5-benzyloxyindole (2.0 g; 8.6 mmol) in DMF (20 mL) was added
dropwise to an ice
cooled suspension of NaH (60% dispersion in mineral oil) (413 mg; 10.32 mmol)
in DMF (30 mL).
The reaction was allowed to warm to room temperature and stirred for 3 hours
until all NaH was
consumed. A solution of iodoacetamide (1.9 g; 10.32 mmol) in DMF (20.mL) was
added and the
reaction left to stir at room temperature for 3 hours. The DMF was removed in
vacuo and the residue
dissolved in ethyl acetate (200 mL). The organic solution was washed with
water until the washings
were clear, brine, dried over anhydrous magnesium sulfate and the solvent
removed in vacuo.
Recrystallisation from ethyl acetate furnished Intermediate C3 as an off white
solid (2.2 g, 87%).
m.p. 204.0-205.0 °C. 1H NMR (500 MHz, DMSO-d6) ~ 4.72 (s, 2H), 5.09 (s,
2H), 6.32 (1H, d, J 3.0),
6.85 (1H, dd, J 8.8, 2.4), 7.12 (1H, d, J2.4), 7.19 (1H, br), 7.24 (2H, m),
7.30 (1H, dd, J7.3, 7.3),
7.37 (2H, dd, J7.4, 7.4), 7.44 (1H, br), 7.45 (2H, d, J7.3).
Intermediate C12: 2-(6-methoxyindol-1-yl) acetamide
Intermediate C12 was prepared as described for Intermediate C2 using 6-
methoxyindole (489 mg,
3.32 mmol), sodium hydride (160 mg, 3.98 mmol) and iodoacetamide (615 mg, 3.32
mmol) in DMF
(50 mL). Two recrystallizations from ethyl acatete yielded intermediate C12 as
a whte solid (252 mg,
37%). m.p. 210.1-211.0 °C. 1H NMR (200MHz, DMSO-D6) 8 7.48 (br s, 1H),
7.41 (d, 8.6Hz, 1H),
7.25 ( br s, 1H), 7.16 (d, 3.2 Hz, 1H), 6.91 (d, 2.0 Hz, 1H), 6.69 (dd, 6.4
Hz, 2.2 Hz, 1H), 6.35 (d, 0.8
Hz, 1H), 4.74 (s, 2H), 3.77 (s, 3H).
Intermediate C13: 7-aza-indol-3-yl acetamide
Intermediate C13 was prepared as described for intermediate C2 using 7-aza-
indole (1.00 g, 8.46
mmol), sodium hydride (406 mg, 10.15 mmol) and iodoacetamide (1.57g, 8.46
mmol) in DMF (100
mL). Two recrystallizations from ethyl acatete yielded Intermediate C5 as a
whte solid (920 mg,
62%). m.p. 180.0-181.0 °C. 1H NMR (200MHz, DMSO-dg) 8 8.21 (dd, 3.lHz,
l.SHz, 1H), 7.95 (dd,
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6.3Hz, l.6Hz, 1H), 7.60 (br s, 1H), 7.48 (d, 3.SHz, 1H), 7.19 (br s, 1H), 7.08
(m, 2H), 6.46 (d, 3.SHz,
1H), 4.88 (s, 3H). MS (EI, m/z) M+= 175.
Intermediate D2:
To a solution of 5-methoxyindole (2.50 g, 17.0 mmol) in 50 mL of anhydrous
diethyl ether cooled to
0 °C was added oxalyl chloride (1.63 mL, 18.68 mmol) and subsequently
warmed to room
temperatue. After 3.5 hours the solution was cooled to 0°C and 50 mL of
an aqueous solution of
ammonium carbonate was added. After stirring overnight the.precipitate was
filtered off and washed
with water, isopropanol and 'diethyl ether to afford the desired acetamide in
78% yield as a bright
yellow solid. m.p. 251.6-252.7 °C iH-NMR (200 MHz, DMSO-d6) 8 11.3
(broad s, 1H), 8.6 (s, 1H),
8.0 (s, 1H), 7.73 (d, 1H, J=2.57 Hz), 7.68 (s, 1H), 7.4 (d, 1H, J=8.81 Hz),
6.8 (dd, 1H, J=2.56;
8.79Hz), 3.8 (s, 3H).
Intermediate Fl: N (indolyl)acetic acid
Indole (1.17g, 10.0 mmol) and h-Bu4NHS03 (339 mg, 1.0 mmol) were dissolved in
benzene (30 mL)
and stirred rapidly with 50% aqueous NaOH (10 mL). After S minutes ethyl
bromoacetate (1.66 mL,
15 mmol) was added and the reaction was stirred for an additional hour. The
aqueous layer was
separated and washed with diethyl ether, acidified and extracted with
methylene chloride under
standard workup conditions to yield intermediate F1 as an off white solid in
72% yield. m.p. 175-178
°C. 1H NMR (500 MHz, DMSO-d6) 8 5.03 (2H, s), 6.47 (1H, d, J3.1), 7.05
(1H, dd, J7.8, 7.8), 7.13
(1H, dd, J8.2, 8.2), 7.35 (1H, d, J3.1), 7.39 (1H, d, J8.2), 7.57 (1H, d,
J7.8), 13.01 (1H, br).
Intermediate F2: N (5-benzyloxyindole) acetic acid
5-Benzyloxyindole (10.00 g, 44.8 mmol) and n-Bu4NHS04 (1.00 g) were
partitioned between toluene
(500 mL) and 50% aqueous NaOH (200 mL). This solution was stirred vigorously
for 30 minutes
before neat ethyl bromoacetate (5.0 mL, 44.8 mmol) was added. After stirring
for an additional 2 h
the mixture was diluted with diethyl ether and water. The aqueous layer was
washed with diethyl
ether before being acidified with 6N HCI. The resulting solid was filtered
off, washed with water, and
dried in vacuo to privide of N (5-benzyloxyindole)acetic acid as an off white
solid (12.21 g, 98 %).
Intermediate G1:
Crude intermediate H1 (l.OOg, 6.21 mmol) was dissolved in THF (100 mL) and
added dropwise to a
cold THF (500 mL) suspension of LiAlH4 (2.70 g, 6.83 mmol). After stirring at
room temperature for
1 hour 2M HCl was added, followed by diethyl ether. The organic layer was
subjected to standard
aqueous workup to provide intermediate G as a clear oil (0.94 g, 94 %).1H NMR
(500 MHz, CDCl3)
8 1.91 (1H, br), 3.79 (2H, t, J 5.3), 4.16 (2H, t, J 5.3), 6.51 (1H, dd, J
3.1, 0.6), 7.10 (1H, d, J 3.1),
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7.14 (1H, ddd, J 7.0, 7.0, 1.0), 7.23 (1H, ddd, J 7.0, 7.0, 1.1), 7.34 (1H,
dd, J 8.3, 0.6), 7.61 (1H, ddd,
J 7.9, 1.0, 1.0).
Intermediate G2:
Crude N (5-benzyloxyindole)acetic acid (12.21 g, 44.2 mmol) was dissolved in
THF (100 mL) and
added dropwise to a cold THF (500 mL) suspension of LiAIH~ (1.78 g, 44.2
mmol). After stirring at
room temperature for 1 hour 2M HCl was added, followed by diethyl ether. The
organic layer was
subjected to standard aqueous workup to provide N (2-hydroxyethyl)-5-
benzyloxyindole as a clear oil
(11.09 g, 94 %). 1H NMR (500 MHz, CDC13) 8 7.51 (d, J=11.8Hz, 1H), 7.50 (d,
J=7.4Hz, 2H), 7.41
(t, J=7.4Hz, 2H), 7.34 (t, J=7.4Hz, 1H), 7.22 (d, J=8.9Hz, 1H), 7.19 (s, 1H),
7.07 (d, J=2.SHz, 1H),
6.99 (d, J=8.9Hz, 1H), 6.42 (d, J=2.SHz, 1H), 5.11 (s, 2H), 4.10 (m, 2H), 3.75
(m, 2H), 2.01 (br s,
1H). MS (EI, m/z) M+=267.
Intermediate H1:
Acetic anhydride (492 ~1, 5.2 mmol) was added to a solution of intermediate
G1(700 mg; 4.34 mmol)
in THF (10 mL) followed by a catalytic amount of DMAP and the reaction left to
stir at room
temperature for 10 minutes. The reaction was diluted with ethyl acetate (50
mL), the organic solution
washed with a saturated solution of sodium bicarbonate (20 mL), water (20 mL),
rinsed with brine (20
mL), dried over anhydrous magnesium sulfate and concentrated in vacuo to yield
intermediate H1 as a
yellow oil (820mg; 93%) which was used without further purification.'H NMR
(500 MHz, CDC13) 8
7.64 (dd, J=0.9, 8.OHz, 1H), 7.36 (dd, J=8.OHz, 1H), 7.23 (dt, J=0.9, 8.OHz,
1H), 7.12 (dt, J=0.9,
8.OHz, 1H), 7.11 (d, J=3.2Hz, 1H), 6.52 (d, H=3.2Hz, 1H), 4.37 (m, 4H), 2.00
(s, 3H).
Intermediate H2:
Crude N (2-hydroxyethyl)-5-benzyloxyindole (11.09 g, 42.1 mmol), acetic
anhydride (4.36 mL, 46.3
mmol), triethylamine (6.50 mL,46.3 mmol), and DMAP (100 mg) were stirred
together in THF for 1
hour before standard aqueous workup provided N (2-acetoxyethyl)-5-
benzyloxyindole as a clear oil
(13.0 g, 99 %).'H NMR (500 MHz, CDCl3) 8 7.51 (d, J=8.lHz, 2H), 7.42 (t,
J=8.lHz, 2H), 7.35 (s,
1H), 7.27 (d, J=8.9Hz, 1H), 7.21 (d, J=2.4Hz, 1H), 7.09 (d, J=3.lHz, 1H), 7.01
(dd, J=2.4, 8.1HZ,
1H), 6.47 (dd, J=0:8, 3.lHz, 1H),5.14(s,2H), 4.37 (t" J=S.OHz 2H), 4.31(t,
J=S.OHz, 2H), 2.02 (s, 3H).
EXEMPLARY PREPARATIVE METHODS FOR FORMULA I TO HI
The following examples provide synthetic methods for preparation of compounds
according
to the invention. Generally, the example number corresponds to the compound
numbers shown in
Tables 2 to 5.
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Example 1: 3-(indol-1-yl)-4-(1H indol-3-yl)-1H pyrrole-2,5-dione
A 1.0M solution of KO'Bu (13.44 mL, 13.44 mmol) was added to a slurry
intermediate B 1
(1.82 g, 8.96 mmol) and intermediate C1 (780 mg; 4.48 mmol) in THF (10 mL).
The reaction slowly
turned red as the reactants started to go into solution. The reaction was
stirred for 3 hours at room
temperature after which time the reaction was quenched by the addition of
concentrated HCl (8 mL)
and diluted with ethyl acetate (100 mL). The organic solution was washed with
water (100 mL),.brine
(100 mL), dried over anhydrous magnesium sulfate and concentrated ih vacuo.
The crude solid was
purified by silica gel chromatography, eluting with 1:1 hexanes/ethyl acetate,
to provide Compound 1
as a red solid (983 mg, 67 %). m.p. 212-213 °C.'H NMR (500 MHz, DMSO-
d6) ~ 6.08 (1H, d, J 8.1),
6.49 (1H, ddd, J 8.1, 8.1, 0.9), 6.75 (1H, dd, J 3.4, 0.7), 6.86 (1H, ddd, J
8.1, 8.1, 1.0), 6.93 (1H, ddd,
J 8.1, 8.1, 1.0), 6.98 (2H, m), 7.33 (1H, d, J 8.1), 7.55 (1H, d, J 3.4), 7.56
(1H, d, J 8.8), 8.04 (1H, d, J
2.9), 11.23 (1H, s), 11.93 (1H, br s).
Example 2: 3-(5-Methoxyindol-3-yl)-4-(indol-1-yl)-1H pyrrole-2,5-dione
Indole was dissolved in THF (25 mL) and treated with a 1.0M THF solution of
KOtBu. This
solution was stirred for 1 hour before ethyl bromoacetate was added. After
stirring hour 3 hours
intermediate D2 was added, followed by a 1.0M THF solution of KOtBu. This
mixture was stirred
over night before being quenced with cone HCl (3 mL) followed by standard
aqueous workup.
Purification by silica gel chromatography, eluting with 3:1 hexane/ethyl
acetate, provided compound
2 as an orange solid (32%). 1H NMR (500 MHz, DMSO-d6) 8 11.83 (s, 1H), 11.18
(s, 1H), 8.05 (d,
J=3.OHz, 1H), 7.55 (d, J=7.8Hz, 1H), 7.50 (d, J=3.3Hz, 1H), 7.19 (d, J=8.7Hz,
1H), 7.07 (d, J=8.2Hz,
1H), 7.01 (t, J=7.2Hz, 1H), 6.93 (t, J=7.2Hz, 1H), 6.69 (d; J=3.3Hz, 1H), 6.50
(dd, J=2.4, 8.7Hz, 1H),
5.49 (d, J=2.3Hz, 1H), 2.93 (s, 3H).
Example 3: 3-(5-Methoxyindol-1-yl)-4-(1H indol-3-yl)-1H pyrrole-2,5-dione
Indole (2.21 g, 18.9 mmol) was dissolved in THF (5 mL) and treated with a 1.0M
solution of
KOtBu (18.9 mL, 18.9 mmol). After stirring for 2 hours neat ethyl bromoacetate
(2.10 mL, 18.9
mmol) was added and the resulting slurry was stirred for an additional 3
hours. Solid intermediate D2
(2.06 g, 9.44 mmol) was added followed by a 1.0M solution of KOtBu (37.8 mL,
37.8 mmol). After
stirring for 16 hours the reaction was quenched with 10 rnL of cone HCl and
subjected to standard
aqueous workup. Purification by silica gel chromatography, eluting with 2.5:1
petroleum
ether/acetone provided compound 4 as an orange/brown solid in 33 % yield. 'H
NMR (200 MHz,
CDC13) 8 8.83 (br s, 1H), 8.00 (br s, 1H), 7.88 (d, J=3.OHz, 1H), 7.39 (d,
J=3.4Hz, 1H), 7.22 (d,
J=8.6Hz, 1H), 7.01 (m, 2H), 6.79 (d, J=9.OHz, 1H), 6.65 (m, 2H), 6.49 (dd,
J=2.5, 9.OHz, 1H), 6.25
(d, J=8.2Hz, 1H), 3.74 (s, 3H).
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Example 4: 3-(5-Methoxy-1H indol-3-yl)-4-(5-methoxyindol-1-yl)-pyrrole-2,5-
dione
5-Methoxyindole (1.00 g, 6.79 mmol) was dissolved in THF (20 mL) and treated
with a 1.0M
solution of KOtBu (6.80 mL, 6.80 mmol). After stirring for 2 hours neat ethyl
bromoacetate (753 ~L,
6.80 mmol) was added and the resulting slurry was stirred for an additional 3
hours. Solid
intermediate D2 (1.48 g, 6.79 mmol) was added followed by a 1.0M solution of
I~OtBu (33.80 mL,
33.80 mmol). After stirring for 3 days the reaction was quenched with 3 mL of
conc HCl and
subjected to standard aqueous workup. Purification by silica gel
chromatography, eluting with 3:2
petroleum ether/acetone provided compound 4 as a deep red solid in 9 % yield.
'H N1VIR (200MHz,
DMSO-d6) 8 11.49 (br s, 1H), 10.43 (br s, 1H), 8.61 (d, J=3.OHz, 1H), 7.95 (d,
J=3.3Hz, 1H), 7.70
(dd, J=5.2, 9.7Hz, 1H), 7.52 (d, J=2.3Hz, 1H), 7.48 (d, J=9.OHz, 1H), 7.25
(dd, J=2.3, 9.OHz, 1H),
7.10 (d, J=0.7Hz, 1H), 7.05 (m, 1H), 6.85 (d, J=0.7Hz, 1H), 6.14 (d, J=2.3Hz,
1H), 4.23 (s, 3H), 4.17
(s, 3H). MS (EI, m/z) M+=387.
Example 5: 3-(5-Benzyloxy-1H indol-3-yl)-4-(indol-1-yl)-pyrrole-2,5-dione
I~O'Bu (1.0M solution in THF; 5.16 mL; 5.16 mmol) was added to a slurry of
intermediate B3
(1.06 8;'3.44 mmol) and Cl (300 mg; 1.72 mmol) in THF (5 mL). The reaction
slowly turned red as
the reactants started to go into solution. The reaction was left for 3 hours
at room temperature before
being quenched by the addition of concentrated HCl (3 mL) and diluted with
ethyl acetate (100 mL).
The organic solution was washed with water (100 mL), brine (100 mL), dried
over anhydrous
magnesium sulfate and concentrated ih vacuo. Purification by silica gel
chromatography using 3:2
hexanes/ethyl acetate as mobile phase yielded Compound 5 as a red solid (689
mg; 92 %). m.p. 217-
220 °C.'H NMR (500 MHz, DMSO-d6) 8 4.03 (2H, s), 5.57 (1H, d, J2.3),
6.60 (1H, dd, J 8.7, 2.3),
6.74 (1H, dd, J3.3, 0.7), 7.22 (1H, d, J8.7), 7.49 (1H, d, J3.3), 7.58 (1H, d,
J7.6), 8.09 (1H, s),
11.19 (1H, s), 11.87 (1H, s).
Example 6: 3-(1=(2-hydoxyethyl)-5-benzyloxyindol-3-yl)-4-(indol-1-yl)-1H
pyrrole-2,5-dione
Intermediate B7 (1.01 g, 0.287 mmol) and Intermediate C1 (0.25 g, 0.144 mmol)
were
suspended in dry THF (100 mL). A 1M solution of potassium test-butoxide (4.31
mL) was added at 0
°C and allowed to stir at room temperature over the weekend. Solvent
was removed in vacuo and the
residue partitioned between ethyl acetate and 1M HCl. The organic layer was
subjected to standard
aqueous workup and further purified using silicon gel chromatography, eluting
with pure ethyl
acetate, to provide Compound 6 as an orange solid (150 mg, 22%). m.p. 205.0-
207.0 °C.'H NMR
(200MHz, DMSO-d6) 8 11.20, (br s, 1H), 8.20 (s, 1H), 7.58 (d, 7.7 Hz, 1H),
7.21, (m, 10H), 6.67 (m,
2H), 5.54, (d, 1.7 Hz, 1H), 5.11 (br s, 1H), 4.23, (m, 2H), 3.99, (s, 2H)..
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Example 7: 3-(1-(2-hydoxyethyl)-5-benzyloxyindol-3-yl)-4-(5-methoxyindol-1-yl)-
1H pyrrole-
2,5-dione
Intermediate B7 (0.5 g, 0.142 mmol) and Intermediate C2 (0.145g, 0.071 mmol)
were
suspended in dry THF (100 mL). A 1M solution of potassium tent-butoxide (2.14
mL) was added at 0
°C and allowed to stir at room temperature over night. Solvent was
removed ire vacuo and the residue
partitioned between ethyl acetate and 1M HCI. The organic layer was subjected
to standard aqueous
workup and further purified using silicon gel chromatography, eluting with
pure ethyl acetate, to
provide Compound 7 as an orange solid was recovered (221 mg, 62%). m.p. 200.0-
201.0 °C. 1H NMR
(200MHz, DMSO-D6) b11.23 (br s, 1H), 8.19 (s, 1H), 7.25 (m, 9H), 6.65 (m, 3H),
5.59 (s, 1H), 5.02
(br s, 1H), 4.27 (br s, 2H), 3.97 (s, 2H), 3.75 (br s, 2H), 3.61 (s, 3H).
Example 8 and 9: 3-(1-(2-hydoxyethyl)-S-benzyloxyindol-3-yl)-4-(6-methoxyindol-
1-yl)-1H
pyrrole-2,5-dione, 8, and 3-(5-benzyloxyindol-3-yl)-4-(6-methoxyindol-1-yl)-
1H pyrrole-2,5-dione, 9.
Intermediate B7 (0.5 g, 0.142 mmol) and Intermediate C3 (0.145g, 0.071 mmol)
was
suspended in dry THF (100 mL). A 1M solution of potassium tent-butoxide (2.14
mL) was added at 0
°C and allowed to stir at room temperature over night. The solvent was
removed ifa vacuo and the
residue partitioned between ethyl acetate and 1M HCI. The organic layer was
subjected to standard
aqueous workup. Purification using silicon gel chromatography, eluting with
2:1 ethyl
acetate/petroleum ether yielded Compound 8 as an orange solid (245 mg, 69%)
along with Compound
9 as an orange solid (24 mg, 8 %). Compound 8: 'H NMR (200MHz, DMSO-d6) 8
11.25 (br s, 1H),
8.17 (s, 1H), 7.30 (m, 8H), 6.65 (m, 4H), 5.63 (s, 1H), 5.00 (t, 5.0 Hz, 1H),
4.36 (br s, 2H), 4.21 (s,
2H), 3.78 (br s, 2H), 3.39 (s, 3H). Compound 9: 1H NMR (200MHz, DMSO-d6) S
11.87 (br s, 1H),
11.21 (br s, 1H), 8.05 (d, 3.0 Hz, 1H), 7.31 (m, 8H), 6.63 (m, 4H), 5.65 (s,
1H), 4.15 (s, 2H).
Example 10: 3-(1-(2-hydoxyethyl)-indol-3-yl)-4-(indol-1-yl)-1H pyrrole-2,5-
dione
KO'Bu (1.0 M solution in THF; 2.73 mL; 2.73 mmol) was added to a slurry of
intermediate
B11 (500 mg; 1.8 mmol) and intermediate C1 (160 mg; 0.91 mmol) in THF (2.5
mL). The reaction
slowly turned red as the reactants started to go into solution. The reaction
was left for 2 hours at room
temperature after which time all C1 had been consumed. The reaction was
quenched by the addition
of concentrated HCl (1.5 mL) and diluted with ethyl acetate (30 mL). The
organic solution was
washed with water (10 mL), brine (10 mL), dried over anhydrous magnesium
sulfate and concentrated
ifa vacuo. Purification by silica gel chromatography using 1:1 hexanes: ethyl
acetate as mobile phase
yielded Compound 10 as a red solid (257 mg; 76%). m.p. 210-212 °C. 1H
NMR (500 MHz, DMSO-
35. d6) ~ 3.73 (2H, dt, J5.3, 5.3), 4.29 (2H, t, J5.3), 4.94 (1H, t, J5.3),
5.99 (1H, d, J 8.1), 6.48 (1H, dd,
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J 8.0, 8.0), 6.72 (1H, d, J 3.3), 6.88 (1H, dd, J 8.2, 8.2), 6.96 (2H, m),
6.99 (1H, d, J 7.3), 7.43 (1H, d,
J8.2), 7.49 (1H, d, J3.3), 7.56 (1H, d, J7.3), 8.14 (1H, s), 11.2 (1H, s).
Example 1 l: 3-(5-benzyloxyindol-1-yl)-4-(1H 5-benzyloxyindol-3-yl)-1H pyrrole-
2,5-dione
KO'Bu (1.0M solution in THF; 1.71 mL; 1.71 mmol) was°added to a slurry
of intermediate B3
(353 mg, 1.14 mmol) and intermediate C3 (160 mg; 0.57 mmol) in THF (2 mL). The
reaction slowly
turned red as the reactants started to go into solution. After stirring for 3
h the reaction was quenched
by addition of concentrated HCl (2 mL) and diluted with ethyl acetate (50 mL).
The organic solution
was washed with water (20 mL), brine (20 mL), dried over anhydrous magnesium
sulfate and
concentrated ih vacuo. Tituration with acetone yielded compound 11 as a red
solid in 68% yield.
Example 12: 3-(5-benzyloxyindol-1-yl)-4-(1H indol-3-yl)-1H pyrrole-2,5-dione
KO'Bu (1.0M solution in THF; 3.21 mL; 3.21 mmol) was added to a slurry of
intermediate Bl
(435 mg; 2.14 mmol) and intermediate C3 (300 mg; 1.07 mmol) in THF (3 mL).
.The reaction slowly
15~ turned red as the reactants started to go into solution. The reaction was
left for 3 hours at room
temperature after which time all intermediate C3 had been consumed by TLC
analysis. The reaction
was quenched by the addition of concentrated HCl (2 mL) and diluted with ethyl
acetate (50 mL).
The organic solution was washed with water (20 mL), brine (20 mL), dried over
anhydrous
magnesium sulfate and concentrated in vacuo. Purification by silica gel
chromatography using 1:1
hexanes/ethyl acetate as mobile phase yielded compound 12 as a red solid (322
mg, 70 %). m.p. 114-
116 °C. 'H NMR (500 MHz, DMSO-d6) 8 5.01 (2H, s), 6.12 (1H, d, J 8.1),
6.52 (1H, dd, J 7.6, 7.6),
6.56 (1H, dd, J8.9, 2.1), 6.65 (1H, d, J3.0), 6.84 (1H, d, J8.9), 6.95 (1H,
dd, J7.6, 7.6), 7.14 (1H, d,
J2.0), 7.34 (6H, m), 7.50 (1H, d, J3.2), 8.00 (1H, d, J2.7), 11.18 (1H, s),
11.90 (1H, br).
Example 13: 3-(indol-1-y1)-4-(1H 5-bromoindol-3-yl)-1H pyrrole-2,5-dione
This structure was prepared according to the procedure described for Example
12 using
intermediate B4 and C1. 1H NMR (200 MHz, DMSO-d6) 8 12.03 (br s, 1H), 11.27
(br s, 1H), 8.05 (s,
1H), 7.59 (d, J=4.SHz, 1H), 7.56 (s, 1H), 7.27 (d, J=8.6Hz, 1H), 7.05 (d,
J=l.BHz, 1H), 6.97 (d,
J=8.6Hz, 1H), 6.86 (t, J=8.6Hz, 2H), 6.79 (d, J=3.3Hz, 1H), 6.02 (d, J=l.BHz,
1H).
Example 14: 3-(indol-1-y1)-4-(1H5-iodoindol-3-yl)-lHpyrrole-2,5-dione
KO'Bu (1.0M solution in THF; 6.9 mL; 6.9 mmol) was added to a slurry of
intermediate BS
(1.5 g; 4.6 mmol) and intermediate C1 (400 mg; 2.3 mmol) in THF (10 mL). The
reaction slowly
turned orange as the reactants started to go into solution. The reaction was
left for 5 hours at room
temperature after which time all Z-ihdol-1 yl-acetamide had been consumed. The
reaction was
quenched by the addition of concentrated HCl (4 mL) and diluted with ethyl
acetate (100 mL). The
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organic solution was washed with water (100 mL), brine (100 mL), dried over
anhydrous magnesium
sulfate and concentrated in vacuo. Purification by silica gel chromatography
using 1:1 hexanes/ethyl
acetate as mobile phase yielded compound 14 as an orange solid (503 mg, 48 %).
m.p. 260-261°C. 1H
NMR (500MHz, DMSO-d6) 8 6.18 (1H, s, ArH), 6.81 (1H, d, J 3.3, ArH), 6.85 91H,
d, J, 7.3, ArH),
6.89 (1H, d, J 8.2, ArH), 7.56 (1H, d, J 3.3, ArH), 7.60 (1H, d, J7.9, ArH),
8.03 (1H, d; J2.9, ArH),
11.25 (1H, s, maleamide NH), 12.04 (1H, br, indole NH).
Example 15: 3-(Indol-1-yl)-4-(5-((3-phenylethynyl)-indol-3-yl)-pyrrole-2,5-
dione
A l .OM THF solution of KO'Bu (1.23 mL, 1.23 mmol) was added to a slurry of
intermediate
B8 (250 mg, 0.82 mmol) and intermediate C1 (71.4 mg, 0.41 mmol) in THF (1.5
mL). The reaction
slowly turned red as the reactants started to go into solution. The reaction
was left for 4 hours at room
temperature until no remaining C1 wasobserved by TLC. The reaction was
quenched by the addition
of concentrated HCl (750 ~L) and diluted with ethyl acetate (30 mL). The
organic solution was
washed with water (20 rnL), brine (20 mL), dried over-anhydrous magnesium
sulfate and concentrated
in vacuo. Puriftcation by silica gel chromatography, eluting with 1:1
hexanes/ethyl acetate, yielded
compound 15 as a red solid (135 mg, 77 %). m.p. 141-144 °C. 1H NMR (500
MHz, DMSO-d6) 8 6.11
(1H, s), 6.79 (1H, d, J3.2), 6.91 (1H, m), 7.01 (2H, dd, J7.0, 7.0), 7.08 (1H,
dd, J 8.3, 1.3), 7.39 (6H,
m), 7.52 (1H, d, J 3.3), 7.58 (1H, d, J 7.8), 8.08 (1H, d, J 1.4), 11.25 (1H,
s), 12.09 (1H, s).
Example 16: 3-(Indol-1-yl)-4-(5-((3 phenethyl)indol-3-yl)-pyrrole-2,5-dione
A 1.0M THF solution of KO'Bu (1.50 mL, 1.50 mmol) was added to a slurry of B9
(307 mg,
1.00 mmol) and C1 (87.0 mg, 0.50 mmol) in THF (1.5 mL). The reaction slowly
turned red as the
reactants started to go into solution. The reaction was left for 16 hours at
room temperature after until
no remaining C1 was observed by TLC.. The reaction was quenched by the
addition of concentrated
HCl (1 mL) and diluted with ethyl acetate (20 mL). The organic solution was
washed with water (15
mL), brine (15 mL), dried over anhydrous magnesium sulfate and concentrated
in,vacuo. Purification
by silica gel chromatography using 1:1 hexanes:ethyl acetate as mobile phase
yielded compound 16 as
a red solid (205 mg, 95 %). m.p. 210-211 °C. 1H NMR (500 MHz, DMSO-d6)
S 2.20 (2H, AB
splitting), 2.28 (2H, AB splitting), 5.68 (1H, s), 6.73 (1H, d, J 8.2), 6.77
(1H, d, J2.8), 6.91 (1H, dd, J
7.6, 7.6), 7.00 (4H, m), 7.14 (1H, dd, J7.3, 7.3), 7.18 (1H, d, J8.2), 7.24
(2H, dd, J7.3, 7.3), 7.53
(1H, d, J3.2), 7.56 (1H, d, J7.8), 8.13 (1H, d, J2.8), 11.19 (1H, s), 11.87
(1H, br).
Example 17: 2-(N benzimidazolyl)-3-(indol-3-yl)-pyrrole-2,5-dione
Benzimidazole (2.56 g, 21.74 mmol) and 3-bromo acetamide (1.5 g, 10.87 mmol)
were added
to dry THF (50 mL). The solution was stirred under nitrogen over night. The
THF is removed in
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vacuo and the resulting solid is partitioned between ethyl acetate and water.
The organic layer, after
removal of the solvent ih vacuo yielded 2-(N-benzimidazolyl)acetamide,
intermediate Kl, as a white
solid. Crude intermediate K1 from above and intermediate Bl (2.20 g, 10.'9
mmol) were suspended in
THF (100 mL) and treated with a 1.0M solution of potassium tart-butoxide (43.5
mL, 43.5 mmol) at 0
°C and stirred for 3 days. Standard workup using ethyl acetate and
aqueous 2M HCl yielded an orange
residue, which was further purified by acetone titration to afford compound 17
as an orange solid (650
mg, 18 %). m.p. 246.4-249.0 °C. 1H NMR (200 MHz, DMSO-d6) b 12.09 (br
s, 1H), 11.41 (br s, 1H),
8.43 (s, 1H), 8.11 (d, J=2.3Hz, 1H), 7.70 (d, 7.9 Hz, 1H), 7.37 (d, J=8.2Hz,
1H), 7.08 (m, 4H), 6.53 (t,
J=7.6 Hz, 1H), 6.08 (d, J=8.lHz, 1H).
Example 18:
Compound 17 (328 mg, 1.0 mmol) was added too a DMF (5 mL) solution of 60% w/w
pariftn/NaH (120 mg, 3 mmol). After stirring for 2 hours Boc20 (273 mg, 1.25
mmol) was added and
the mixture as stirred for 16 hours. Standard aqueous/ethyl acetate workup,
followed by silica gel
chromatography, eluting with 1:1 petroleum ether/ehtyl acetate, yieled
compound 18 was a yellow
solid (165 mg, 39%).
Example 19:
Compound 17 (296 mg, 0.903 mmol), DMAP (5 mg), triethylamine (138 p,L, 0.990
mmol),
and acetyl chloride (64 pL, 0.900 mmol) were stirred together in DMF (5 mL)
for 16 hours. Standard
aqueous/ethyl acetate workup and purification by silica gel chromatography,
eluting with 2:1
petroleum ether/acetone, provided compound 19 as a red solid. 1H NMR (200 MHz,
DMSO-d6)
8 11.40 (br s, 1H), 8.45 (s, 1H), 8.14 (s, 1H), 8.08 (d, J=7.8Hz, 1H), 7.68
(d, J=8.OHz, 1H), 7.20-6.98
(m, 4H), 6.78 (t, J=7.8Hz, 1H), 6.30 (d, J=7.8Hz, 1H), 2.68 (s, 3H).
Example 20:
Compound 17, triethylamine, and phenylisocyanate were refluxed in THF for 24
hours. The
solvent was removed in vacuo and the resulting solid purified by silica gel
chromatography, eluting
with 2:1 hexane/acetone, to provide compound 20 as a yellow solid in 78%
yield. 1H NMR (200 MHz,
DMSO-d6) 8 11.66 (br s, 1H), 10.56 (s, 1H), 8.66 (s, 1H), 8.46 (s, 1H), 8.12
(d, J=8.3Hz, 1H), 7.68 (d,
J=7.8Hz, 3H), 7.41 (t, J=8.OHz, 1H), 7.23-6.99 (m, SH), 6.71 (t, J=7.lHz, 1H),
6.29 (d, J=8.lHz, 1H).
Example 21: 3-(N benzimidazolyl)-4-(5-benzyloxyindol-3-yl)-pyrrole-2,5-dione
Intermediate B3 (1.18 g, 4 mmol), intermediate Kl (700 mg, 4 mmol), KZC03
(4.42 g, 32
mmol), and CETAB (102 mg, 0.28 mmol) are suspended in dry benzene (150 mL).
This mixture was
refluxed for 4 days, removing water using a Dean Stark apparatus. The solvent
was removed ih vacuo
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and the resulting solid was partitioned between ethyl acetate and 2M aqueous
HC1. The organic layer
was washed with water, dried over anhydrous MgS04, altered and dried down
under reduced
pressure. The resulting red solid was further purifified by two silica gel
chromatography columns, the
first eluting with 2.5:1 petroleum ether/acetone, and the second eluting with
1:1 petroleum
ether/acetone, to provide compound 21 as an orange solid (90 mg, 5 %). 1H NMR
(500 MHz, DMSO-
d6) 8 12.05 (br s, 1H), 8.34 (s, 1H), 8.19 (s, 1H), 7.74 (d, J=8.OHz, 1H),
7.26 (m, 9H), 6.65 (dd, J=2.3,
8.8Hz, 1H), 5.53 (d, J=2.2Hz, 1H), 3.97 (s, 2H), 3.08 (s, 3H).
Example 22:
Intermediate B1 (0.25 g, 1.43 mmol) and Intermediate C13 (0.58 g, 2.86 mmol)
were
suspended in dry THF (100 mL). A 1M solution ofpotassium tart-butoxide (4.30
mL) was added at 0
°C and allowed to stir at room temperature over night. Solvent was
removed in vacuo and the residue
partitioned between ethyl acetate and 1M HCl. The organic layer was subjected
to standard aqueous
worlcup and further purified using silicon gel chromatography, eluting with
1:1 ethyl
acetate/petroleum ether, to provide Compound 21 as an orange solid was
recovered (59 mg, 12%). 1H
NMR (200MHz, DMSO-d6) 8 11.99 (br s, 1H), 11.28 (br s, 1H), 8.14 (d, J=2.8Hz,
1H), 8.01 (m, 2H),
7.65 (d, J=3.6Hz, 1H), 7.33 (d, J=8.lHz, 1H), 7.04 (m, 2H), 6.76 (d, J=2.9Hz,
1H), 6.77 (m, 1H), 5.92
(d, J=8.lHz, 1H).
Example 23:
Compound 22 (79 mg, 0.238 mmol), DMAP (29 mg, 0.238 mmol) and Boc20 (55 ~,L,
0.238
mmol) were stirred together in THF (25 mL) for 16 hours. Standard
aqueous/ethyl acetate workup
and purification by preparative TLC, eluting with 3:1 petroleum ether/ethyl
acetate, provided
compound 23 as a red solid (35 mg, 34 %).
Example 24:
Compound 1 (1.6 g; 4.89 mmol) was partially solubilised in dichloromethane
(150 mL).
TMSOTf (1.08 mL, 5.87 mmol) was added and the reaction left to stir at room
temperature for 2
hours. The reaction was split into 3 x 50 mL portions and each portion treated
as follows. The
reaction was diluted with ethyl acetate (500 mL), washed with a saturated
solution of sodium
bicarbonate (250 mL), water (250 mL), brine (250 mL), dried over anhydrous
magnesium sulfate and
concentrated in vacuo. Purification by silica gel chromatography, eluting with
1:1 hexaneslethyl,
provided compound 23 as a dark purple solid (1.41 g, 88 %). m.p. >300
°C. 'H NMR (500 MHz,
DMSO-d6) ~ 3.64 (1H, dd, J 9.9, 16.5), 3.91 (1H, dd, J 2.5, 16.5), 5.10 (1H,
dd, J 2.5, 9.9), 7.16 (3H,
m), 6.98 (1H, dd, J7.6, 7.6), 6.78 (2H, m), 7.26 (1H, d, J7.4), 7.44 (1H, d,
J7:6), 7.98 (1H, d, J2.7),
10.57 (1H, s), 11.81 (1H, br).
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Example 25 and 26:
Compound 24 (500 mg, 1.50 mmol) was suspended in ethanol (50 mL) and treated
with 4
portions of NaBH4 (170 mg, 4.5 minol) over 4 hours. The reaction was quenched
with water and
5 diluted with ethyl acetate. The organic layer was washed with aqueous NH4C1
followed by saturated
aqueous NaHC03, and water, dried over anhydrous MgS04,, filtered and volatiles
removed under
reduced pressure, providing a 1:1 mix of regeoisomers, compounds 25 and 26 in
89% combined yield.
Example 27:
10 ' Compound 27 was placed in a quartz tube, dissolved in acetonitrile, and
placed in front of a
150W light bulb for 16 hrs. Removal of the solvent under reduced pressure
provided compound 27 as
an off white solid.'H NMR (200 MHz, DMSO-d6) 8 11.30 (s, 1H), 7.90 (d,
J=2.4Hz, 1H), 7.80 (s,
1H), 7.37 (d, J=7.8Hz, 1H), 7.28 (d, J=7.lHz, 1H), 7.17-6.99 (m, 4H), 6.84 (t,
J=7.lHz, 1H), 5.29 (dd,
J=8.0, 9.4Hz, 1H), 4.96 (dd, J=8.0, 8.9Hz, 1H), 3.88 (d, J=17.8Hz, 2H), 3.77
(m, 1H).
Example 29:
Compound 24 (1.00 g, 3.05 mmol) was dissolved in THF (50 mL) and treated with
Boc20
(772 ~,I,, 3.36 mmol) and DMAP (5 mg). The solution was stirred for 10 minutes
before standard
aqueous/ethyl acetate workup and purification by silica gel chromatography,
eluting with 3:1
hexane/ethyl acetate, provided compound 29 as a red solid (530 mg, 41 %).
Example 30 and 31:
Compound 29 (3.45 mg, 8.09 mmol) was dissolved in EtOH (150 mL) and treated
with
NaBH4 (1.80 g, 48.5 mmol). After stirring for 3 hours the reation mixture was
quenched with water
and diluted with ethyl acetate. The organic layer was washed with aqueous
NH4C1 and worked up as
usual provided a 3:1 mixture of regeoisomers. Purification by silica gel
chromatography, eluting with
3:1 hexane/ethyl acetate, provided compound 30 (1.09 g, 31%, as a light brown
solid, and compound
31 (340 mg, 10%) as an off white solid.
Compound 30: 'H NMR (200 MHz, DMSO-d6) 8 8.25 (s, 1H), 8.20 (s, 1H), 8.04 (d,
J=7.2Hz, 1H),
7.41-7.20 (m, 4H), 7.06 (t, J=7.2Hz, 1H), 6.84 (t, J-7.4Hz, 1H), 6.30 (d,
J=12.OHz, 1H), 6.14 (d,
J=9.9Hz, 1H), 5.29 (dd, J=5.4, 10.5Hz, 1H), 3.91 (dd, J=5.4, 16.6Hz, 1H), 3.67
(dd, J=10.5, 16.6Hz,
1H), 1.63 (s, 9H).
Compound 31: 'H NMR (200 MHz, DMSO-d6) 8 8.14 (d, J=3.lHz, 1H), 8.01 (d,
J=7.OHz, 1H), 7.95
(d, J=8.4Hz, 1H), 7.33 (m, 3H), 7.13 (t, J=7.6Hz, 1H)~ 6.95 (t, J=7.3Hz, 1H),
6.77 (d, J=9.9Hz, 1H),
5.51 (d, J=9.9Hz, 1H), 5.37 (dd, J=6.8, 9.9Hz, 1H), 3.93 (dd, J=6.8, 18.SHz,
1H), 3.62 (dd, J=9.9,
18.SHz, 1H), 1.64 (s, 9H).
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Example 32: Major isomer
Compound 30 (50 mg, 0.12 mmol) andp-toluenesulfonic acid (5 mg) were dissolved
in
methylene chloride (4 mL) and treated with phenyl selenol (89 ~L, 0.84 mmol).
The solution was
stirred for 1 hour before the volatiles were removed under reduced pressure
and the resulting solid
was purified by silica gel chromatography, eluting with 19:1 methylene
chloride/methanol, to provide
compound 32 as a light brown solid (39 mg, 80%). 1H NMR (200 MHz, DMSO-d6) 8
8.21 (s, 1H),
8.04 (d, J=7.7Hz, 1 H), 7.94 (s, 1 H), 7.41-7.21 (m, 4H), 7.12 (t, J=7. 9Hz, 1
H), 6. 92 (t, J=7.2Hz, 1 H),
5.40 (dd, J=7.2, 10.2Hz, 1H), 4.92 (d, J=1.8.2Hz, 1H), 3.98 (d, J=18.2Hz, 1H),
3.86-3.70 (m, 2H), 1.64
(s, 9H). MS (FAB, m/z) M+1=414.
Example 33: Minor isomer
Compound 31 (100 mg, 0.24 mmol) andp-toluenesulfonic acid (5 mg) were
dissolved in
methylene chloride (5 mL) and treated with phenyl selenol (74 ~L, 0.69 mmol).
The solution was
stirred for 1 hour before the volatiles were removed under reduced pressure
and the resulting solid
was purified by silica gel chromatography, eluting with 19:1 methylene
chloridelmethanol, to provide
compound 32 as a light brown solid (75 mg, 79%).
Example 34:
Compound 3 was partially solubilized in dichloromethane and treated with neat
TMSOT~
The mixture was stirred for 16 hours. The solvent was removed ih vacuo and the
resulting residue
dissolved in ethyl acetate (10 mL). The organic solution was washed with
saturated sodium
bicarbonate solution (10 mL), water (10 mL), brine (10 mL), dried over
anhydrous magnesium sulfate
and concentrated in vacuo. PuriEcation by silica gel chromatography, eluting
with 1:1 hexanes/ethyl
acetate, provided compound 34 as a dark purple solid. m.p. 248.4-250.5
°C. 1H NMR (200 MHz,
DMSO-d6) 8 11.70 (s, 1H), 10.53 (s, 1H), 7.94 (d, J=2.6Hz, 1H), 7.43 (d,
J=7.lHz, 1H), 7.13 (m, 2H),
6.90 (d, J=l.BHz, 1H), 6.76 (d, J=8.6Hz; 1H), 6.59 (dd, J=2.4, 8.6Hz, 1H),
5.00 (d, J=7.SHz, 1H), 3.89
(d, J=7.SHz, 1H), 3.69 (s, 3H), 3.66 (m, 1H). MS (EI, m/z) M+=357.
Example 35:
Compound 5 (20 mg; 0.046 mmol) was partially solubilized in dichloromethane (2
mL). Neat
boron trifluoride diethyl etherate (11 ~,L; 0.055 mmol) added and the reaction
left to stir at room
temperature for 5 hours. The solvent was removed in vacuo and the resulting
residue dissolved in
ethyl acetate (10 mL). The organic solution was washed with saturated sodium
bicarbonate solution
(10 mL), water (10 mL), brine (10 mL), dried over anhydrous magnesium sulfate
and concentrated in
vacuo. Purification by silica gel chromatography, eluting with 1:1
hexanes/ethyl acetate, provided
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compound 35 as a dark purple solid (13 mg; 65 %). mp 258.0-261.0 °C. 1H
NMR (500 MHz, DMSO-
d6) b 11.69 (br s, 1H), 10.49 (br s, 1H), 7.92 (s, 1H), 7.31 (m, 6H), 7.07 (d,
J=8.8 Hz, 1H), 6.86 (d,
J=8.1 Hz, 1H), 6.32 (s, 1H), 6.29 (d, J=5.7 Hz, 1H), 5.31 (t, J=4.9 Hz, 1H),
5.08 (d, J=11.6 Hz, 1H),
5.06 (d, J=11.6Hz, 1H), 5.15 (m, 1H), 3.65 (m, 1H), 3.63 (s, 3H).
Example 36:
Compound 6 (0.5 g, 1.05 mmol) was dissolved in CHZCl2 (50 mL). Neat BF3~Et02
(0.216
mL) was added via syringe at -78 °C. The temperature was allowed to
come to room temperature over
night. Solvent was removed in vacuo, and the resulting solid was suspended in
methanol and placed in
the freezer for 2 hours. Filtration and washing with cold methanol yielded
Compound 36 as a purple
solid (320 mg, 64%). m.p. 238.6-240.5 °C. 1H NMR (SOOMHz, DMSO-d6) ~
10.50 (br s, 1H), 7.98 (s,
1H), 7.48 (m, 3H), 7.41 (m, 2H), 7.36 (m, 1H), 7.10 (d, J=9.OHz, 1H), 6.98 (m,
2H), 6.72 (m, 2H),
5.29 (dd, J=5.2, 4.7Hz, 1H), 5.08 (dd, J=15.9 Hz, 11.6Hz, 2H), 4.89 (t,
J=5.2Hz, 2H), 4.27, (m, 3H),
3.70, (m, 3H).
Example 37:
Compound 36 (15 mg, 0.0314 mmol), DMAP (3.84 mg, 0.0314 mmol), and NEt3 (4.81
~.L)
were dissolved in dry THF (25 mL). Acetoxy acetyl chloride (3.4 uL) was added
via syringe and the
reaction was allowed to stir over night at room temperature. The THF was
removed on a rotary
evaporator and the resulting solid worked up in ethyl acetate and water.
Purification of the crude solid
by silicon gel chromatography, eluting with 1:1 ethyl acetatelpetroleum ether,
provided compound 37
as a light yellow solid (8.2 mg, 45 %). 1H NMR (SOO.IMHz, DMSO-d6) 8 10.52 (br
s, 1H), 7.99 (s,
1H), 7.45 (m, 6H), 7.14 (d, J=9.OHz, 1H), 6.97 (m, 2H), 6.72 (m, 2H), 5.32
(dd, J=4.9, S.OHz, 1H),
5.10 (d, J=16.2 Hz, 2H), 4.52 (m, 4H), 4.40 (m, 2H), 4.23 (dd, J=4.9, 11.9Hz,
1H), 3.72 (rn, 1H), 2.50
(m, 2H), 2.02 (s, 3H).
Example 38:
Compound 38 was prepared as described for Compound 36, using Compound 7 (50.0
mg,
0.0986 mmol) and BF3~Et02 (0.0224 mL) in CHZCIz (50 mL) to provide Compound 38
as a purple
solid (6.3 mg, 12%). m.p. 248.0-249.0 °C. 1H NMR (SOOMHz, DMSO-d6) b
10.41 (br s, 1H), 7.94 (s,
1H), 7.43 (m, 6H), 7.07 (d, J=9.OHz, 1H), 6.71 (d, J=8.4Hz, 1H), 6.56 (m, 2H),
5.27 (dd, J=5.5,
4.3Hz, 1H), 5.07 (q, J=11.6Hz, 2H), 4.88 (t, J=5.3Hz, 1H), 4.28 (m, 3H), 3.65
(m, 3H), 3.65 (s, 3H).
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Example 39:
Compound 39 was prepared as described for Compound 36, using Compound 8 (55
mg, 0.108
mmol) and BF3~Et02 (226 ~L) in CHzCIz (50 mL) to provide Compound 38 as a
purple solid (3.6 mg,
7%). m.p. >300.0 °C. 1H NMR (500MHz, DMSO-d6) S 10.49 (br s, 1H), 7.98
(s, 1H), 7.42 (m, 6H),
7.10 (d, J=9.0 Hz, 1H), 6.86 (d, J=8.0 Hz, 1H), 6.30 (m, 2H), 5.31 (dd, J=4.6,
5.1 Hz, 1H), 5.08 (dd,
J=11.7, 15.4 Hz, 2H), 4.88 (t, J=6.8Hz, 2H), 4.27 (m, 2H), 4.16 (dd, J=4.6,
11.7 Hz, 1H), 3.68 (m,
3H), 3.62 (s, 3H).
Example 40:
Compound 10 (95 mg, 0.26 mmol) was partially solubilised in dichloromethane
(15 mL).
Trimethylsilyl trifluoromethanesulfonate (65 ~L, 0.30 mmol) was added and the
reaction left to stir at
room temperature fox 1 h. Standard aqueous workup and purification by silica
gel chromatography,
eluting with ethyl acetate, provided compound 40 as a dark purple solid (25
mg, 26 %). m.p. 280-282
°C. 1H NMR (500 MHz, DMSO-d6) 3.64 (1H, dd, J 10.0, 16.5), 3.71 (2H,
dt, J 5.3, 5.3), 3.92 (1H, dd,
J2.5, 16.5), 4.30 (m, 2H), 4.89 (1H, t, J5.3), 5.10 (1H, dd, J2.5, 10.0), 6.78
(1H, dd, J7.5, 7.5), 6.98
(1H, dd, J7.6, 7.6), 7.19 (2H, m), 7.26 (1H, d, J7.3), 7.55 (1H, d, J7.8),
8.02 (1H, s), 10.58 (s, 1H).
Example 41:
Compound 12 (20 mg, 0.046 mmol) was partially solubilised in dichloromethane
(2 mL),
boron trifluoride diethyl etherate (11 ~L, 0.055 mmol) added and the reaction
left to stir at room
temperature for 5 hours. The solvent was removed in vacuo and the resulting
residue dissolved in
ethyl acetate (10 mL). The organic solution was washed with saturated sodium
bicarbonate solution
(10 mL), water (10 mL), brine (10 mL), dried over anhydrous magnesium sulfate
and concentrated in
vacuo. Purification by silica gel chromatography, eluting with 1:1
hexanes/ethyl acetate, provided
compound 41 as a dark purple solid. m.p. 238-242 °C.'H NMR (500 MHz,
DMSO-d6) 8 3.64 (1H,
dd, J 10.0, 16.7, CH), 3.87 (1H, dd, J 2.8, 16.8, CH), 5.03 (2H, s, benzylic
CHZ), 5.11 (1H, dd, J 2.8,
9.9, CH), 6.68 (1H, dd, J2.5, 8.7, ArH), 6.75 (1H, d, J 8.7, ArH), 6.98 (1H,
d, J2.2, ArH), 7.11 (1H,
d, J 7.5, ArH), 7.16 ( 1 H, dd, J 7.6, 7.6, ArH), 7.30 ( 1 H, dd, J 7.2, 7.2,
ArH), 7.3 7 (2H, dd, J 7.4, 7.4, 2
ArH), 7.42 (3H, m, 3 ArH), 7.93 (1H, d, J2.7, ArH), 10.49 (1H, s, maleamide
NH), 11.75 (1H, br,
indole NH). HRMS (m/z) Cal'd for CZ~H19N3O3 433.14264. Found 433.1420.
Example 42:
A solution of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (122 mg, 0.540 mmol)
in 1,4-
dioxan (5 mL) was added dropwise to a solution of compound 24 (80.0 mg, 0.240
mmol) in 1,4-
dioxan (5 mL) and the reaction left to stir at room temperature for 2 hours.
The solvent was removed
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in vacuo and the resulting residue purified by silica gel chromatography,
eluting with 1:1 -
hexanes/ethyl acetate, to provide compound 41 as a dark purple solid (43 mg,
54 %).
mp >300°C. HRMS (mlz) Cal'd for CZOH11N3O3 325.0851. Found 325.0855.
Example 43 and 44:
Compound 42 was dissolved in methanol and treated with 3 portions of sodium
borohydride,
at 15 minute intervals. The solution was stirred for an additional 60 minutes
before being quenched
with 1M HCl and extracted with ethyl acetate. The crude product contained a
1:1 mixture of
regeoisomers, one of which was isolated pure by tituration with acetone. Major
Isomer: 1H NMR (200
MHz, DMSO-d6) 8 11.39 (s, 1H), 8.56 (s, 1H), 8.07 (d, J=2.4Hz, 1H), 7.91 (d,
J=8.6Hz, 1H), 7.58 (d,
J=7.7Hz, 1H), 7.51 (s, 1H), 7.49 (d, J=7.7Hz, 1H), 7.25 (d, J=8.OHz, 1H), 7.17-
7.07 (m, 3H), 6.56 (d,
J=lO.OHz, 1H), 6.39 (d, J=lO.OHz, 1H). MS (EI, m/z) M+=327.
Example 47:
Compound 2 (50 mg, 0.14 mmol) was stirred with TMSOTf (32.4 pL, 0.168 mmol) in
CHzCl2
(5 mL) for 3 days. Removal of volatiles and purification by silica gel
chromatography, eluting with
3:2 hexanes\ethyl acetate, yielded a deep purple solid in 67% yield. 1H-NMR
(200 MHz, DMSO-d6) ~
11.82 (s, 1H), 10.93 (s, 1H), 7.97 (d, J=3.SHz, 1H), 7.68 (s, 1H), 7.45
(m,.lH), 7.38 (m, 1H), 7.30 (d,
J=7.8Hz, 1H), 7.06 (m, 3H), 3.95 (s, 3H).
Example 48:
Compound 34 (50 mg, 0.14 mmol) was stirred with TMSOTf (32.4 p,L, 0.168 mmol)
in
CH2Clz (5 mL) for 3 days. Removal of volatiles and purification by silica gel
chromatography,
eluting with 3:2 hexanes\ethyl acetate, yielded compound 48 as deep purple
solid in 42% yield. m.p.
>300 °C. 1H NMR(200 MHz, DMSO-d6) 8 11.94 (s, 1H), 10.96(s, 1H), 8.01
(d, J=2.6Hz, 1H), 7.58 (d,
J=7.2Hz, 1H), 7.40 (d, J=9.lHz, 1H), 7.29 (t, J=9.lHz, 1H), 7.26 (d, J=9.lHz,
1H), 7.17 (t, J=9.lHz,
1H), 6.95 (d, J=2.6Hz, 1H), 6.69 (dd, J=2.6, 9.lHz, 1H), 3.77 (s, 3H). MS (EI,
m/z) M+=355.
Example 49:
Compound 4 (392 mg, 1.00 mmol) was dissolved in methylene chloride (25 mL) and
treated
with neat TMSOTf (215 pL, 1.11 mmol). After stirring over night the volatiles
were removed in
vacuo and the resulting material purified by silica gel chromatography,
eluting with 2:1 petroleum
ether/acetone, to provide compound 49 as a purple solid in 31 % yield. 1H NMR
(200 MHz, DMSO-
d6) 8 11.70 (br s, 1H), 11.00 (br s, 1H), 7.94 (s, 1H), 7.60 (s, 1H), 7.26 (d,
J=9.OHz, 1H), 7.04 (d,
J=9.OHz, 1H), 6.98 (d, J=2.4Hz, 1H), 6.78 (dd, J=2.5, 9.OHz, 1H), 3.93 (s,
3H), 3.76 (s, 1H).
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Example 50:
Compound 47 (50 mg, 0.141 mmol), triethylamine (22 ~L, 0.154 mmol), DIC (24
~L, 0.154
mmol), Boc-Gly-OH (27 mg, 0.154 mmol), and DMAP (2 mg) were dissolved in THF
and refluxed
for 16 hours. Volatiles were removed under reduced pressure and purification
by silica gel
5 chromatography, eluting with 25:1 methylene chloride/methanol, followed by
preparative TLC,
eluting with 25:1 methylene chloride/methanol, provided compound 46 as a deep
red solid in 46
yield. 1H NMR (200 MHz, DMS,O-d6) 8 11.20(br s,lH), 8.24(s,lH), 8.14(d,
J=9.0Hz,lH), 7.72 (s,lH),
7.52-7.30(m, 3H), 7.21(d, J=9.OHz, 1H), 7.11(m, 2H), 4.47(d, J=S.OHz, 2H),
3.96(s, 3H), 1.43 (s, 9H).
10 Example 51:
Compound 47 (25 mg, 0.070), triethylamine (11 ~L, 0.077 mmol), acetoxyacetyl
chloride (8
~,L, 0.077 mmol), and DMAP (1 mg) were dissolved in THF and stirred for 16
hour. Volatiles were
removed under reduced pressure and purification by preparative TLC, eluting
with 15:1 methylene
chloride/methanol, provided compound 46 as a deep red solid in 34 % yield. 1H
NMR (200 MHz,
15 DMSO-d6) 8 11.22 (br s, 1H), 8.24 (s, 1H), 8.16 (d, J=8.lHz, 1H), 7.82 (s,
1H), 7.52 (m, 2H), 7.30 (d,
J=9.3Hz, 1H), 7.12 (m, 2H), 5.48 (s, 2H), 4.01 (s, 3H), 2.19 (s, 3H).
Example 52:
A solution of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (44.5 mg; 0.20 mmol)
in 1,4-dioxan
20 (5 mL) was added dropwise to a solution of compound 35 (38.6 mg; 0.089
mmol) in 1,4-dioxan (5
mL) and the reaction left to stir at room temperature for 2 hours. The solvent
was removed in vacuo
and the resulting residue purified by silica gel chromatography, eluting with
1:1 hexanes/ethyl acetate,
provided compound 52 as a dark purple solid (20 mg; 52 %). m.p. 197-200
°C. 1H NMR (500 MHz,
DMSO-d6) 8 5.27 (2H, s, benzylic CHZ), 7.02 (1H, ddd, J7.5, 7.5, 0.9, ArH),
7.07 (1H, ddd, J7.0,
25 7.0, 1.3, ArH), 7.11 (1H, d, J 8.8, ArH), 7.26 (1H, d, J 8.8, ArH), 7.38
(5H, m, ArH), 7.52 (2H, d, J
7.0, ArH), 7.72 (1H, d, J 0.7, ArH), 7.96 (1H, d, J 2.8, ArH), 10.92 (1H, s;
maleamide NH), 11.83
(1H, br, indole NH). HRMS (m/z) Cal'd for CZ~H1~N303 431.1270. Found 431.1273.
Example 53:
30 Compound 20 (20 mg) was placed in a quartz tube and dissolved in
acetonitrile. This solution
was placed in front of a 150W bulb for 48 h. Compound 53 precipitated from
solution, was removed
by filtration, and washing with aceonitrile, to provide compound 53 as a deep
red solid (2 mg, 10%).
'H NMR (200 MHz, DMSO-d6) 8 13.40 (br s, 1H), 11.48 (br s, 1H), 9.34 (d,
J=8.lHz, 1H), 8.78 (d,
J=8.lHz, 1H), 7.79 (d, J=8.lHz, 1H), 7.70 (t, J=8.lHz, 1H), 7.62-7.55 (m, 4H),
7.41 (t, J=8.lHz, 1H).
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Example 54:
Compound 19 (100 mg) was placed in a quartz tube and dissolved in
acetonitrile. This
solution was placed in front of a 150W light bulb for 16 hours. Compound 53
precipitated from
solution and was removed by filtration, washing with aceonitrile, to provide
compound 53 as an
orange solid (28 mg, 28%). 1H NMR (200 MHz, DMSO-d6) 8 11.75 (br s, 1H), 9.36
(d, J=7.9Hz, 1H),
8.92 (d, J=7.7Hz, 1H), 8.13 (d, J=8.3Hz, 1H), 8.02 (d, J=7.7Hz, 1H), 7.70-7.52
(m, 4H), 3.15 (s, 3H).
Example 55:
Compound 18 (20 mg) was placed in a quartz tube and dissolved in acetonitrile.
This solution
was placed in front of a 150W light bulb for 48 hours. Compound 53
precipitated from solution and
was removed by filtration, and titurated with methylene chloride, to provide
compound 55 as a deep
red solid (10 mg, 50%). MS,(EI, mlz) M+=426.
Example 56: Synthesis of 3-(1H 5-bromoindol-3-yl)-1H pyrrole-2,5-dione
Oxalyl chloride (445 uL, 5.10 mmol) ) was added to a THF (25 ) solution of 5-
bromoindole
(1.00 g, 5.10 mmol). After stirring at room temperature for 3 hours, volatiles
were removed under
reduced pressure. Solid acetamide (903 mg, 15.3 mmol) was added to the
resulting solid, and the
mixture was dissolved in THF (10 mL). After stirring for 3 hours, a 1M
solution of I~OtBu (15.3 mL,
15.3 mmol) was added. The resulting, deep purple solution was stirred
overnight, quenched with conc
HZS04 (1 mL), stirred for 30 minutes before being diluted with water (20 mL)
and ethyl acetate. The
aqueous layer was washed with ethyl acetate and the combined organic layers
were dried over
anhydrous MgS04, filtered, and the solvent removed under reduced pressure.
Purification by silica
gel chromatography, eluting with 3:1 to 1:1 petroleum ether/ethyl acetate,
provided compound 56 in
45% yield. Compound 56 could be further purified, where necessary, by
recrystallization from
methanol. m.p. 270.5-271.0 °C.'H NMR (200MHz, acetone-d6) d 11.28 (br
s, 1H), 9.66 (br s, 1H),
8.48 (d, J=l.SHz, 1H), 8.15 (s, 1H), 7.53 (d, J=8.5Hz, 1H), 7.42 (d, J=8,5Hz,
1H), 6.82 (s, 1H).
Example 57: Synthesis of 3-(N acetyl-5-bromoindol-3-yl)-1H pyrrole-2,5-dione
Compound 56 (80 mg, 0.275 mmol), triethylamine (42 pL, 0.3 mmol), DMAP (5 mg),
and
actetic anhydride (30 pL, 0.3 mmol) were stirred together in THF (5 mL) over
night. Standard
aqueous worlcup and purification by silica gel chromatography, eluting with
3:1 petroleum ether/ethyl
acetate, provided compound 57 in 51% yield, as a yellow solid. m.p. 271.0-
272.9 °C. 1H NMR (200
MHz, DMSO-d6) 8 11.07 (br s, 1H), 8.49 (s, 1H), 8.25 (d, J=8.5Hz, 1H), 8.18
(s, 1H), 7.56 (d,
J=8.5Hz, 1H), 7.24 (s, 1H), 2.70 (s, 3H).'3C NMR (74.5MHz, DMSO-d6) 8 172.4,
172.2, 169.7,
136.3, 133.9, 131.0, 129.1, 128.6, 123.0, 117.8, 117.3, 109.4.
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Example 58: 3-(N (N,N dimethylcarbamoyl)-5-bromoindol-3-yl)-1H pyrrole- 2,5-
dione
Compound 56 (80 mg, 0.275 mmol), triethylamine (42 ~L, 0.3 mmol), DMAP (5 mg),
and
N,N dimethylcarbamoyl chloride (26 p,L, 0.3 mmol) were stirred together in THF
(5 mL) over night.
Standard aqueous workup and recrystallization from ethyl acetate, provided
compound 58 in 21%
yield, as a yellow solid. The filtrate was approximately 95% pure. m.p. 287.0-
288.0 °C. 1H NMR
(200 MHz, DMSO-d6) 8 10.98 (br s, 1H), 8.43 (s, 1H), 8.22 (s, 1H), 7.57 (d,
J=8.7Hz, 1H), 7.48 (d,
J=8.7Hz, 1H), 3.02 (s, 6H).
Example 59: Synthesis of 3-(N benzoyl-5-bromoindol-3-yl)-1H pyrrole-2,5-dione
Compound 56 (80 mg, 0.275 mmol), triethylamine (42 ~L, 0.3 mmol), DMAP (5 mg),
and
benzoyl chloride (52 p,I,, 0.3 mmol) were stirred together in THF (5 mL) over
night. Standard
aqueous workup and recrystallization from ethyl acetate, provided compound 59
in 51% yield, as a
yellow solid.
Example 60: 3-(N (p-toluenesulfonyl)-5-bromoindol-3-yl)-1H pyrrole-2,5-dione
Compound 56 (50 mg, 0.172 mmol), DMAP (44 mg, 260 mmol), andp-toluenesulfonyl
chloride (33 mg, 0.172 mmol) were refluxed in THF (5 mL) for 48 hours.
Standard aqueous workup
and purification by silica gel chromatography, eluting with 4:1 petroleum
ether/ethyl acetate, provided
compound 60 in 91 % yield, as a yellow solid. m.p. 262.0-263.8 °C. 1H
NMR (200 MHz, DMSO-d6) 8
11.13 (s, 1H), 8.54 (s, 1H), 8.23 (s, 1H), 7.93 (d, J=8.2Hz, 2H), 7.93 (d,
J=8.8Hz, 1H), 7.58 (dd,
J=2.1, 8.8Hz, 1H), 7.40 (d, J=8.2HZ, 2H), 7.32 (s, 1H), 2.31 (s, 3H).
Example 61: 3-(N (1-naphthylenesulfonyl)-5-bromoindol-3-yl)-1H pyrrole-2,5-
dione
Compound 56 (146 mg, 0.50 mmol), triethylamine (77 ~L, 0.55 mmol), DMAP (10
mg), and
1-naphthylenesulfonyl chloride (113mg, 0.5 mmol) were refluxed in THF (5 mL)
for 48 hours.
Standard aqueous workup and purification by silica gel chromatography, eluting
with 3:1 petroleum
ether/ethyl acetate, provided compound 61 in 73% yield, as an off white
solid.'H NMR (200 MHz,
DMSO-d6) 8 11.14 (br s, 1H), 8.78 (s, 1H), 8.54 (d, J=7.4Hz, 1H), 8.50 (d,
J=8.2Hz, 1H), 8.36 (d,
J=8.2Hz, 1H), 8.20 (s, 1H), 8.08 (d, J=7.8Hz, 1H), 7.79-7.63 (m, 4H), 7.49 (d,
J=9.OHz, 1H), 7.30 (s,
1H).'3C NMR (74.SMHz, DMSO-d6) 8 172.4, 172.2, 169.7, 136.3, 133.9, 131.0,
129.1, 128.6, 123.0,
117.8, 117.3, 109.4.
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Example 62: 3-(N (2-naphthylenesulfonyl)-5-bromoindol-3-yl)-1H pyrrole-2,5-
dione
Compound 56 (146 mg, 0.50 mmol), triethylamine (77 q,L, 0.55 mmol), DMAP (10
mg), and
2-naphthylenesulfonyl chloride (113mg, 0.5 mmol) were refluxed in THF (5 mL)
for 48 hours.
Standard aqueous workup and purification by silica gel chromatography, eluting
with 3:1 petroleum
ether/ethyl acetate, provided compound 62 in 48% yield, as an off white
solid.'H NMR (200 MHz,
DMSO-d6) 8 11.14 (br s, 1H), 8.96 (s, 1H), 8.64 (s, 1H), 8.26-8.22 (m, 2H),
8.10 (d, J=9.OHz, 1H),
8.05-7.98 (m, 1H), 7.99 (dd, J=2.0, 8.7Hz, 1H), 7.74-7.65 (m, 3H), 7.57 (dd,
J=1.7, 9.OHz, 1H), 7.31
(s, 1H).'3C NMR (74.SMHz, DMSO-d6) 8 172.4, 172.2, 135.8, 135.1, 132.9, 132.7,
131.5, 130.5,
130.3, 130.2, 129.8, 129.6, 129.3, 128.8, 128.3, 128.0, 123.8 (2 C's), 121.1,
117.8, 115.2, 110.5.
Example 63: Synthesis of 3-(N dansyl-5-bromoindol-3-yl)-1H pyrrole-2,5-dione
Compound 56 (146 mg, 0.50 mmol), triethylamine (77 q,L, 0.55 mmol), DMAl' (10
mg), and
dansyl chloride (202 mg, 0.75 mmol) were refluxed in THF (5 mL) for 48 hours.
Standard aqueous
workup and purification by silica gel chromatography, eluting with 3:1
petroleum ether/ethyl acetate,
provided compound 63 in 42% yield, as an off white solid. 'H NMR (200 MHz,
DMSO-d6) 8 11.14
(br s, 1H), 8.78 (s, 1H), 8.57 (d, J=7.2Hz, 1H), 8.54 (d, J=7.2Hz, 1H), 8.24
(s, 1H), 8.11 (d, J=8.8Hz,
1H), 7.74 (J, J=8.4Hz, 2H), 7.66 (d, J=8.OHz, 1H), 7.58 (d, J=8.6Hz, 1H), 7.53
(d, J=8.8Hz, 1H), 7.33
(s, 1H), 7.20 (d, J=7.7Hz, 1H), 2.74 (s, 6H).'3C NMR (74.SMHz,. DMSO-d6) 8
171.8, 171.6, 151.6,
135.3, 132.4, 132.1, 130.8, 129.8, 129.2, 128.5, 128.4, 128.1, 127.9, 123.4,
123.3, 117.0, 115.9, 115.4,
114.3, 113.1, 109.1, 44.4.
Example 64: Synthesis of 3-(1-H 5-benzyloxyindol-3-yl)-1H pyrrole-2,5-dione
Compound 64 was prepared in a manner similar manner to compound 56, using 5-
benzyloxyindole (2.00 g, 8.96 mmol), oxalyl chloride (0.82 mL, 9.41 mmol),
acetamide (1.70 g, 28.7
mmol), and a 1M solution of I~O'Bu (45 mL, 44.8 mmol). Standard workup and
purification first by
silica gel chromatography, eluting with 3:1 petroleum etherlethyl acetate,
followed by
recrystallization from MeOH, provided compound 64 as an orange solid in 53%
yield. ~H NMR (200
MHz, DMSO-d6) 8 11.92 (s, 1H), 10.72 (s, 1H), 8.31 (d, J=3.OHz, 1H), 7.53-7.31
(m, 7H), 6.95 (dd,
J=3.0, 8.OHz, 1H), 6.82 (s, 1H), 5.22 (s, 2H). 13C NMR (74.SMHz, DMSO-d6) 8
173.5, 173.3, 154.3,
139.4, 137.7, 131.6, 131.2, 128.4, 127.7, 126.2, 114.8, 113.6, 113.3, 105.3,
103.9, 69.9.
Example 65: Synthesis of 3-(N acetyl-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Compound 64 (50 mg, 0.164 mmol), triethylamine (23 ~L, 0.164 mmol), acetic
anhydride (16
~,L, 0.164 mmol), and DMAP (2 mg) were' stirred together in THF (5 mL) over
night. Standard
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aqueous workup and purification by silica gel chromatography, eluting with 3:1
petroleum ether/ethyl
acetate, provided compound 65 in 78 % yield, as a yellow solid.
Example 66: Synthesis of 3-(N benzoyl-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Compound 64 (50 mg, 0.164 mmol), triethylamine (23 ~,L, 0.164 mmol), benzoyl
chloride (19
~L, 0.164 mmol), and DMAP (2 mg) were stirred together in THF (5 mL) over
night. Standard
aqueous workup and purification by silica gel chromatography, eluting with 4:1
petroleum ether/ethyl
acetate, provided compound 66 in 51% yield, as a yellow solid. 1H NMR (500
MHz, DMSO-d6) 8
10.99 (s, 1H), 8.27 (s, 1H), 8.24 (d, J=9.OHz, 1H), 7.80 (d, J=7.4Hz, 1H),
7.73 (t, J=7.4Hz, 1H), 7.63
(t, J=7.4Hz, 2H), 7.60 (d, J=2.2Hz, 1H), 7.51 (d, J=7.4Hz, 2H), 7.41 (t,
J=7.4Hz, 2H), 7.33 (t,
J=7.4Hz, 1H), 7.29 (s, 1H), 7.18 (dd, J=2.3, 9.OHz, 1H), 5.28 (s, 2H).'3C NMR
(74.8MHz, DMSO-
d6) 8 172.7, 172.5, 168.0, 156.2, 137.2, 136.7, 133.3, 132.6, 131.8, 130.2,
129.3, 128.83, 128.79,
128.4, 127.83, 127.79, 122.3, 117.0, 155.2, 110.2, 105.0, 70Ø
Example 67: 3-(N (2,4-dimethoxybenzoyl)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Compound 64 (76 mg, 0.25 mmol), triethylamine (35 ~L, 0.25 mmol), 2,4-
dimethoxybenzoyl
chloride (30 ~L, 0.3 mmol), and DMAP (2 mg) were stirred together in THF (5
mL) over night.
Standard aqueous workup and recrystallization from methanol, provided compound
67 in 62% yield,
as a yellow solid. 1H NMR (500 MHz, DMSO-d6) 8 10.97 (s, 1H), 8.24 (d,
J=9.OHz, 1H), 8.09 (s,
1H), 7.56 (d, J=l.4Hz, 1H), 7.52 (d, J=9.9Hz, 1H), 7.50 (d, J=8.6Hz, 1H), 7.40
(t, J=7.5Hz, 2H), 7.33
(t, J=7.SHz, 1H), 7.25 (s, 1H), 7.14 (dd, J=2.4, 9.OHz, 1H), 6.79 (d, J=2.2Hz,
1H), 6.73 (dd, J=2.2,
9.OHz, 1H), 5.26 (s, 2H), 3.89 (s, 3H), 3.73 (s, 3H). 13C NMR (74.8MHz, DMSO-
d6) 8 172.6, 172.3,
166.4, 163.4, 157.9, 156.0, 137.2, 136.7, 131.9, 131.3, 129.6, 128.8, 128.4,
127.8, 127.7, 122.0, 116.6,
115.4, 115.0, 110.0, 106.1, 104.9, 98.84, 69.9, 55.9, 55.7.
Example 68: 3-(N (3,4-dimethoxybenzoyl)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Compound 64 (76 mg, 0.25 mmol), triethylamine (35 ~L, 0.25 mmol), 3,4-
dimethoxybenoyl
chloride (30 ~L, 0.3 mmol), and DMAP (2 mg) were stirred together in THF (5
mL) over night.
Standard aqueous worlcup and recrystallization from methanol, provided
compound 68 in 74% yield,
as a yellow solid.'H NMR (200 MHz, DMSO-d6) 8 11.04 (s, 1H), 8.42 (s, 1H),
8.20 (d, J=9.OHz,
1H), 7.51-7.22 (m, 8H), 7.21 (s, 1H), 7.15 (d, J=9.OHz, 1H), 5.29 (s, 2H),
3.90 (s, 3H), 3.84 (s, 3H).
Example 69: 3-(N (N Boc-2-aminoacetyl)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Compound 64 (22 mg) was dissolved in THF (5 mL) and refluxed with N Boc-Gly-
OPf (50
mg), and DMAP (2 mg) for 48 hrs. The solvent was removed ifa vacuo and
purified by silica gel
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chromatography, eluting with 3:1 hexane/ethyl acetate, to provide a 3:1
inseparable mixture of
compound 69 and compound 64 (34 mg). 1H NMR(200 MHz, DMSO-d6) 811.10 (s,lH),
8.60(s,lH),
8.40(m,2H), 8.08(m,lH), 7.60-7.35(m,SH), 7.20(s,lH), 4.50(m 2H), 3.50(s,2H),
1.42(s, 9H).
5 Example 70: Synthesis of 3-(N tosyl-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Compound 64 (100 mg, 0.314 mmol), triethylamine (48 ~L, 0.346 mmol), DMAP (10
mg),
and p-toluenesulfonyl chloride (66 mg, 0.346 mmol) were refluxed in THF (5 mL)
for 48 hours.
Standard aqueous workup and purification by silica gel chromatography, eluting
with 3:1 petroleum
ether/ethyl acetate, provided compound 70 in 86% yield, as an off white solid.
1H NMR (200 MHz,
10 DMSO-d6) 8 11.11 (s, 1H), 8.52 (s, 1H), 7.90 (d, J=8.2Hz, 2H), 7.88 (d,
J=9.4Hz, 1H), 7.52-7.31 (m,
8H), 7.27 (s, 1H), 7.12 (dd, J=2.2, 9.lHz, 1H), 5.19 (s, 2H), 2.31 (s, 3H).
13C NMR (125.7MHz,
DMSO-d6) 8 172.5, 172.3, 156.1, 146.2, 137.0, 136.3, 133.3, 130.5, 129.7,
128.7, 128.4, 127.8, 127.7,
127.0, 122.8, 115.5, 114.3, 113.6, 111.0, 105.1, 70.0, 21Ø
15 Example 71: 3-(N (4-nitrobenzenesulfonyl)-5-benzyloxyindol-3-yl)-1H pyrrole-
2,5-dione
Compound 64 (50 mg, 0.157 mmol), triethylamine (24 ~,L, 0.157 mmol), DMAP (2
mg), and
4-nitrobenzenesulfonyl chloride (38 mg, 0.173 mmol) were refluxed in THF (5
mL) for 48 hours.
Standard aqueous workup and purification by silica gel chromatography, eluting
with 4:1 to 1:1
petroleum ether/ethyl acetate, provided compound 71 in 27% yield, as a light
yellow solid. 1H NMR
20 (200 MHz, DMSO-d6) 8 11.16 (s, 1H), 8.53 (s, 1H), 8.33 (br s, 4H), 7.92 (d,
J=9.OHz, 1H), 7.53-7.41
(m, 4H), 7.36 (d, J=7.6Hz, 2H), 7.32 (s, 1H), 7.15 (dd, J=1.2, 9.OHz, 1H),
5.20 (s,'2H). 13C NMR
(74.8MHz, DMSO-d6) 8 172.5, 172.2, 156.4, 151.1, 141.1, 137.0, 136.0, 129.4,
128.9, 128.7, 128.4,
127.9, 127.8, 125.3, 123.5, 120.1, 115.8, 114.3, 112.0, 105.3, 70Ø
25 Example 72: 3-(N (3-nitrobenzenesulfonyl)-5-benzyloxyindol-3-yl)-1H pyrrole-
2,5-dione
Compound 64 (50 mg, 0.157 mmol), triethylamine (24 ~,L, 0.157 mmol), DMAP (2
mg), and
3-nitrobenzenesulfonyl chloride (38 mg, 0.173 mmol) were refluxed in THF (5
mL) for 48 hours.
Standard aqueous worlcup and purification by silica gel chromatography,
eluting with 4:1 to 1:1
petroleum etherlethyl acetate, provided compound 72 in 42% yield, as a light
yellow solid.
Example 73: Synthesis of 3-(N benzyl-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Step 1: 5-Benzyloxyindole, intermediate A1 (500 mg, 2.24 mmol), was dissolved
in benzene
(15 mL) and treated with a 50% sodium hydroxide solution (5 mL) in the
presence of
tetrabutylammonium hydrogensulfate (76 mg, 0.22 mmol) for 30 minutes, followed
by the addition of
benzyl bromide (0.40 mL, 3.36 mmol). After stirring for 48 hours, standard
aqueous workup
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provided 800 mg of crude material. Purification by silica gel chromatography,
eluting with 10:1
hexanes\ethyl acetate, afforded clean product in 50 % yield. 1H NMR (200 MHz,
CDCl3) ~ 7.30 (m,
15H), 6.59 (d, J=2.2Hz, 1H), 5.22 (m, 4H).
St-ep 2: Compound 73 was prepared from N benzyl-5-benzyloxyindole in a similar
fashion as
that described for compound 56 using N Benzyl-5-benzyloxyindole (0.96 mmol),
oxalyl chloride
(83.5 ~L, 0.96 mmol), acetamide (0.15 g, 2.55 mmol), and 1M THF solution of
KOtBu (4.1 mL, 4.07
mmol). The reaction mixture was stirred for 3 days at room temperature.
Standard workup and
purification using silica gel chromatography, eluting with 3:1 hexanes/ethyl
acetate, provided
compound 73 as a yellow solid in 21% overall yield. 1H NMR (200 MHz, DMSO-d6)
b 10.77 (s, 1H),
8.50 (s, 1H), 7.38 (m, 12H), 6.95 (dd, J=2.15; 8.97Hz, 1H), 6.86 (s, 1H), 5.51
(s, 2H), 5.20 (s, 2H).
Example 74: 3-(N (3-pyridinylmethylene)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Step l: 5-Benzyloxyindole (500 mg, 2.24 mmol) was dissolved in THF (20 mL) and
treated
with 1M KOtBu in THF (4.48 mL, 4.48 mmol). After stirring for 3 hours, 3-
(bromomethyl)pyridine
hydrobromide (1.13 g, 4.48 mmol) was added and the reaction mixture was
stirred for an additional
24 hrs. Standard aqueous workup and puriftcation by silica gel chromatography,
eluting with 4:1
hexane/ethyl acetate, provided N (3-pyridinylmethylene)-5-benzyloxyindole (421
mg, 68 %).'H
NMR (200 MHz, CDC13) 8 8.50 (s, 2H), 7.55-7.25 (m, SH), 7.54-7.04 (m, SH),
6.84 (dd, J=2.3,
8.9Hz, 1H), 6.49 (d, J=3.2Hz, 1H), 5.24 (s, 2H), 5.00 (s, 2H). MS (EI, m/z)
M+=314.
Step 2: Compound 74 was prepared from N (3-pyridinylmethylene)-5-
benzyloxyindole in a
similar fashion as that described for compound 56 using N (2,3-
dimethoxybenzyl)-5-benzyloxyindole
(100 mg, 0.318 mmol), oxalyl chloride (27 ~L, 0.318 mmol), acetamide (56 mg,
0.954 mmol), and
1M THF solution of KOtBu (1.60 mL, 1.60 mmol). The reaction mixture was
stirred for 3 days at
room temperature. Standard workup and purification using silica gel
chromatography, eluting with
3:1 hexanes/ethyl acetate, provided compound 74 as a yellow solid in 4% yield.
MS (EI, m/z)
M+=409.
Example 75: 3-(N (2,3-dimethoxybenzyl)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Step 1: 5-Benzyloxyindole (500 mg, 2.24 mmol) was dissolved in THF (20 mL) and
treated
with 1M KOtBu in THF (2.24 mL, 2.24 mmol). After stirring for 3 hours, 3,5-
dimethoxybenzyl
bromide (460 mg, 2.46 mmol) was added and the reaction mixture was stirred for
an additional 24 hrs.
Standard aqueous workup and purification by silica gel chromatography, eluting
with 5:1
hexane/ethyl acetate, provided N (2,3-dimethoxybenzyl)-5-benzyloxyindole (465
mg, 76 %). 'H
NMR (200 MHz, CDC13) 8 7.46 (d, J=7.9Hz, 2H), 7.38 (t, J=7.9Hz, 2H), 7.32 (t,
J=7.9Hz, 1H), 7.23
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(s, 1H), 7.18 (m, 2H), 7.09 (d, J=2.SHz, 1H), 6.91 (m, 1H), 6.45 (d, J=2.OHz,
1H), 6.34 (m, 1H), 6.23
(d, J=2.SHz, 1H), 5.19 (s, 2H), 5.08 (s, 2H), 3.80 (s, 6H).
Step 2: Compound 75 was prepared from N (2,3-dimethoxybenzyl)-5-
benzyloxyindole in a
similar fashion as that described for compound 56 using N (2,3-
dimethoxybenzyl)-5-benzyloxyindole
(250 mg, 0.56 mmol), oxalyl chloride (49 wL, 0.56 mmol), acetamide (99 mg,
1.68 mmol), and 1M
THF solution of KOtBu (2.80 mL, 2.80 mmol). The reaction mixture was stirred
for 3 days at room
temperature. Standard workup and purification using silica gel chromatography,
eluting with 3:1
hexanes/ethyl acetate, provided compound 75 as a yellow solid in 56% yield. MS
(EI, m/z) M+=444.
Example 76: 3-(N (3-fluorobenzyl)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-dione
Step 1: 5-Benzyloxyindole (500 mg, 2.24 mmol) was dissolved in THF (20 mL) and
treated
with 1M KOtBu in THF (2.24 mL, 2.24 mmol). After stirring for 3 hours, 3-
fluorobenzyl bromide
(274 ~,L, 2.24 mmol) was added and the reaction mixture was stirred for an
additional 24 hrs.
Standard aqueous workup and purification by silica gel chromatography, eluting
with 5:1
hexanes/ethyl acetate, provided N (2-fluorobenzyl)-5-benzyloxyindole (325 mg,
44 %). 'H NMR (200
MHz, CDC13) S 7.55-7.29 (m, SH), 7.28-7.05 (m, 3H), 7.04-6.81 (m, SH), 6.49
(dd, J=0.7, 3.lHz, 1H),
5.26 (s, 2H), 5.10 (s, 2H). MS (EI, m/z) M+=331.
Step 2: Compound 76 was prepared from N (2-fluorobenzyl)-5-benzyloxyindole in
a similar
fashion as that described for compound 56 using N 2-(fluorobenzyl)-5-
benzyloxyindole (250 mg, 0.75
mmol), oxalyl chloride (65 ~,L, 0.75 mmol), acetamide (132 mg, 2.25 mmol), and
1M THF solution of
KOtBu (3.75 mL, 3.75 mmol). The reaction mixture was stirred for 3 days at
room temperature.
Standard workup and purification using silica gel chromatography, eluting with
3:1 hexanes/ethyl
acetate, provided compound 76 as a yellow solid in 43% overall yield'H NMR
(200 MHz, DMSO-d6)
8 10.78 (s, 1H), 8.52 (s, 1H), 7.46 (m, 3H), 7.36 (m, 4H), 7.09 (m, 2H), 7.05
(s, 1H), 7.00 (m, 2H),
6.87 (s, 1H), 5.53 (s, 2H), 5.20 (s, 2H). 13C NMR (DMSO-d6) 8 172.2, 171.9,
153.4, 138. MS (EI,
m/z) M+=426.
Example 77: 3-(N (4-fluorobenzyl)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-dione
S-tep 1: 5-Benzyloxyindole (500 mg, 2.24 mmol) was dissolved in THF (20 mL)
and treated
with 1M KOtBu in THF (2.24 mL, 2.24 mmol). After stirring for 3 hours, 4-
fluorobenzyl bromide
(274 p,L, 2.24 mmol) was added and the reaction mixture was stirred for an
additional 24 hrs.
Standard aqueous workup and purification by silica gel chromatography, eluting
with 5:1
hexane/ethyl acetate, provided N (4-fluoro)benzyl-5-benzyloxyindole (455 mg,
61 %). 1H NMR (200
MHz, CDC13) 8 7.60-7.38 (m, SH), 7.23 (d, J=3.lHz, 1H), 7.18-6.92 (m, 7H),
6.50 (d, J=3.lHz, 1H),
5.23 (s, 2H), 5.12 (s, 2H). MS (EI, m/z) M~=331.
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Step 2: Compound 77 was prepared from N (4-fluorobenzyl)-5-benzyloxyindole in
a similar
fashion as that described for compound 56 using N (4-fluorobenzyl)-5-
benzyloxyindole (250 mg, 0.75
mmol), oxalyl chloride (65 ~L, 0.75 mmol), acetamide (132 mg, 2.25 mmol), and
1M THF solution of
KOtBu (3.75 mL, 3.75 mmol). The reaction mixture was stirred for 3 days at
room temperature.
Standard workup and purification using silica gel chromatography, eluting with
3:1 hexanes/ethyl
acetate, provided compound 77 as a yellow solid in 42% overall yield. 1H NMR
(200 MHz, DMSO-
d6) ~ 10.77 (s, 1H), 7.56-7.44 (m, 10H) 7.13 (t, J=8.9Hz, 2H), 6.96 (dd,
J=2.2, 6.8Hz, 1H), 6.86 (s,
1H), 5.50 (s, 2H), 5.20 (s, 2H). MS (EI, m/z) M+=426.
Example 78: 3-(N (methylenephthalimido)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
. Step 1: 5-Benzyloxyindole (500 mg, 2.24 mmol) was dissolved in THF (20 mL)
and treated
with 1M KOtBu in THF (2.24 mL, 2.24 mmol). After stirring for 3 hours, N
(bromomethyl)
phthalimide (538 mg, 2.24 mmol) was added and the reaction mixture was stirred
for an additional 24
hrs. Standard aqueous workup and purification by silica gel chromatography,
eluting with 3:1
hexane/ethyl acetate, provided N (methylenephthalimido)-5-benzyloxyindole (650
mg, 76 %). 1H
NMR (200 MHz, CDC13) 8 7.83 (d, J=5.5 Hz, 1H), 7.81 (d, J=S.SHz, 1H), 7.72-
7.65 (m, 2H), 7.45 (m,
1H), 7.42 (d, 3.OHz, 2H), 7.40-25 (m, 2H), 7.09 (d, J=2.2Hz, 1H), 7.00 (dd,
J=2.2, 9.OHz, 1H), 6.49
(d, J=3.2Hz, 1H), 5.91 (s, 2H), 5.07 (s, 2H). MS (EI. m/z) M+=382.
Step 2: Compound 78 was prepared from N (methylenephthalimido)-5-
benzyloxyindole in a
similar fashion as that described for compound 56 using N
(methylenephthalimido)-5-
benzyloxyindole (250 mg, 0.654 mmol), oxalyl chloride (75 mL, 0.654 mmol),
acetamide (116 mg,
1.96 mmol), and 1M THF solution of KOtBu (3.30mL, 3.30 mmol). The reaction
mixture was stirred
for 3 days at room temperature. Standard workup and purification using silica
gel chromatography,
eluting with 2:1 hexanes/ethyl acetate, provided compound 78 as a yellow solid
in 32% yield.1H,
NMR (200 MHz, DMSO-d6) 8 10.80 (s, 1H), 9.50 (br t, 1H), 8.50 (s, 1H), 7.75
(m, 2H), 7.60-7.25 (m,
10H), 7.06 (dd, J=0.7, 8.3Hz, 1H), 6.86 (s, 1H), 5.64 (br d, J=5.3Hz, 2H),
5.21 (s, 2H).
Example 79: ' 3-(N (2-naphthylmethylene)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Step 1: 5-Benzyloxyindole (500 mg, 2.24 mmol) was dissolved in THF (20 mL) and
treated
with 1M KOtBu in THF (2.24 mL, 2.24 mmol). After stirring for 3 hours, 2-
(bromomethyl)naphthylene (246 mg, 2.24 mmol) was added and the reaction
mixture was stirred for
an additional 24 hrs. Standard aqueous workup and purification by silica gel
chromatography, eluting
with 5:1 hexane/ehtyl acetate, provided N (2-naphthylmethylene)-5-
benzyloxyindole.
1H NMR (200 MHz, CDCl3) b 7.95-7.81 (m, 4H), 7.64-7.50 (m, 4H), 7.7.47 (m,
1H), 7.43 (m, 1H),
7.39 (m, 1H), 7.30 (dd, J=4.0, 9.8Hz, 1H), 7.21 (d, J=2.7Hz, 1H), 7.07 (td,
J=2., 8.9Hz, 1H0, 6.98 (s,
1H), 6.65 (d, J=3.lHz, 1H0, 6.65 (d, J=11.6Hz, 2H), 5.18 (d, J=14.6Hz, 2H). MS
(EI, m/z) M+=363.
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Step 2: Compound 79 was prepared from N (2-naphthylmethylene)-5-
benzyloxyindole in a
similar fashion as that described for compound 56 using N (2-naphthylmethyl)-5-
benzyloxyindole
(250 mg, 0.690 mmol), oxalyl chloride (60 ~L, 0.69 mmol), acetamide (122 mg,
2.07 mmol), and 1M
THF solution of KOtBu (3.45 mL" 3.45 mmol). The reaction mixture was stirred
for 3 days at room
temperature. Standard workup and purification using silica gel chromatography,
eluting with 3:1
hexanes/ethyl acetate, provided compound 79 as a yellow solid in 49 % yield.'H
NMR (200 MHz,
DMSO-d6) & 10.77 (s, 1H), 8.57 (s, 1H), 7.82 (m, 3H), 7.6-7.40 (m, 6H), 7.39-
7.24 (m, 6H), 6.93 (dd,
J=2.1, 9.OHz, 1H), 6.88 (s, 1H), 5.68 (s, 2H), 5.12 (s, 2H). MS (EI, m/z)
M+=458.
Example 80 Synthesis of 3-(N (cyclohexylmethylene)-5-benzyloxyindol-3-yl)-1H
pyrrole-
2,5-dione
Step 1: 5-Benzyloxyindole (500 mg, 2.24 mmol) was dissolved in THF (20 mL) and
treated
with 1M KOtBu in THF (2.24 mL, 2.24 mmol). After stirring for 3 hours,
(bromomethyl)cyclohexane
(312 ~L, 2.24 mmol) was added and the reaction mixture was stirred for an
additional 24 hrs.
Standard aqueous worlcup and purification by silica gel chromatography,
eluting with 5:1
hexane/ethyl acetate, provided N (2-naphthylmethylene)-5-benzyloxyindole as a
white solid (475 mg,
66 %). 1H NMR (200 MHz, CDC13) 8 7.56 (m, 2H), 7.51-7.38 (m, 3H), 7.25 (m,
2H), 7.07 (dd, J=2.5,
6.4Hz, 1H), 7.02 (dd, J=2.5, 6.4Hz, 1H), 6.46 (d, J=2.SHz, 1H), 5.17 (s, 2H),
3.93 (d, J=4.lHz, 2H,
1.91 (m, 1H), 1.88-1.60 (m, SH), 1.56 (m, 3H), 1.11 (m, 2H). MS (EI, m/z)
M+=319.
Step 2: Compound 80 was prepared from N (cyclohexylmethylene)-5-
benzyloxyindole in a
similar fashion as that described for compound 56 using N (2-naphthylmethyl)-5-
benzyloxyindole
(250 mg, 0.78 mmol), oxalyl chloride (68 ~,L, 0.78 mmol), acetamide (138 mg,
2.34 mmol), and 1M
THF solution of KOtBu (3.90 mL, 3.90 mmol). The reaction mixture was stirred
for 3 days at room
temperature. Standard workup and purification using silica gel chromatography,
eluting with 3:1
hexanes/ethyl acetate, provided compound 80 as a yellow solid in 41% yield. 1H
NMR (200 MHz, .
CDCl3) 8 8.23 (s, 1H), 7.52-7.24 (m, 8H), 7.02 (dd, J=2.4, 9.OHz, 1H), 6.42
(s, 1H), 5.13 (s, 2H), 3.93
(d, J=7.lHz, 2H), 1.85 (m, 1H), 1.79-.1.57 (m, 4H), 1.30-0.95 (m, 6H). MS (EI,
m/z) M+'=415.
Example 81: Synthesis of 3-(N octyl-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-
dione
Step 1: N Octyl-5-benzyloxyindole'was prepared by general method A. 5-
Benzyloxyindole
(500 mg, 2.24 mmol) was dissolved in THF (20 mL) and treated with 1M KOtBu in
THF (3.81 mL,
3.8 mmol). After stirring for 3 hours, 1-bromooctane (0.39 mL, 2.24 mmol) was
added and the
reaction mixture was stirred for an additional 24 hrs. Standard aqueous workup
and purification by
silica gel chromatography, eluting with 3:1 hexane/ethyl acetate, provided N
octyl-5-benzyloxyindole.
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1H NMR (200 MHz, CDC13) 8 7.68 (m, 7H), 7.35 (m, 2H), 6.78 (d, J=2.9Hz, 1H),
5.40 (s, 2H), 4.23
(t, J=7.OHz, 2H), 2.05 (broad t, J=6.SHz, 2H), 1.62 (broad s, 10H), 1.31 (t, J-
-6.2Hz, 3H)
Step 2: Compound 81 was prepared from N octyl-5-benzyloxyindole in a similar
fashion as
that described for compound 56. To a solution of N octyl-5-benzyloxyindole in
TI-~' was added oxalyl
5 chloride (0.12 mL, 1.4 mmol), stirred for 24 hours followed by addition of
acetamide (0.22 g, 3.74
mmol), and 1M THF solution of I~OtBu (5.95 mL, 5.95 mmol). The reaction
mixture was stirred for 3
days at room temperature. Standard workup and purification using silica gel
chromatography, eluting
with 5:1 hexanes/ethyl acetate, yielded compound 81 as an orange solid in 30%
overall yield.
'H NMR (200 MHz, DMSO-d6) 8 10.73 (s, 1H), 8.33 (s, 1H), 7.41 (m, 7H), 7.01
(d, J=2.4Hz, 1H),
10 6.80 (s, 1H), 5.21 (s, 2H), 4.23 (t, J=2.7Hz, 2H), 1.72 (m, 2H), 1.21 (s,
10H), 0.83 (m, 3H).
Example 82: 3-(N (2-hydroxyethyl)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-dione
Step 1: 5-Benzyloxyindole (10.00 g, 44.8 mmol) and h-Bu~NHSO~ (1.00 g) were
partitioned
between toluene (500 mL) and 50% aqueous NaOH (200 mL). This solution was
stirred vigerously
15 for 30 minutes before neat ethyl bromoacetate (5.0 mL, 44.8 mmol) was
added. After stirring for an
additional 2 hours the mixture was diluted with diethyl ether and water. The
aqueous layer was
washed with diethyl ether before being acidified with 6N HCI. The resulting
solid was filtered off,
washed with water, and dried ifa vacuo to privide of N (5-
benzyloxyindole)acetic acid as an off white
solid (12.21 g, 98 %).
20 Step 2: Crude N (5-benzyloxyindole)acetic acid (12.21 g, 44.2 mmol) was
dissolved in THF
(100 mL) and added dropwise to a cold THF (500 mL) suspension of LiAlH4 (1.78
g, 44.2 mmol).
After stirring at room temperature for 1 hour 2M HCl was added, followed by
diethyl ether. The
organic layer was subjected to standard aqueous workup to provide N (2-
hydroxyethyl)-5-
benzyloxyindole in yield as a clear oil (11.09 g, 94 %).
25 1H NMR (500 MHz, CDC13) 8 7.51 (d, J=11.BHz, 1H), 7.50 (d, J=7.4Hz, 2H),
7.41 (t, J=7.4Hz, 2H),
7.34 (t, J=7.4Hz, 1H), 7.22 (d, J=8.9Hz, 1H), 7.19 (s, 1H), 7.07 (d, J=2.SHz,
1H), 6.99 (d, J=8.9Hz,
1H), 6.42 (d, J=2.SHz, 1H), 5.11 (s, 2H), 4.10 (m, 2H), 3.75 (m, 2H), 2.01 (br
s, 1H). MS (EI, m/z)
M+=267.
Step 3: Crude N (2-hydroxyethyl)-5-benzyloxyindole (11.09 g, 42.1 mmol),
acetic anhydride
30 (4.36 mL,, 46.3 mmol), triethylamine (6.50 mL,46.3 mmol), and DMAP (100 mg)
were stirred
together in THF for 1 hour before standard aqueous workup provided N (2-
acetoxyethyl)-5-
benzyloxyindole in yield as a clear oil (13.0 g, 99 %). 1H NMR (500 MHz,
CDC13) ~ 7.51 (d,
J=8.lHz, 2H), 7.42 (t, J=8.lHz, 2H), 7.35 (s, 1H), 7.27 (d, J=8.9Hz, 1H), 7.21
(d, J=2.4Hz, 1H), 7.09
(d, J=3.lHz, 1H), 7.01 (dd, J=2.4, 8.1HZ, 1H), 6.47 (dd, J=0.8, 3.lliz, 1H),
5.14 (s, 2H), 4.37 (t"
35 J=S.OHz, 2H), 4.31 (t, J=S.OHz, 2H), 2.02 (s, 3H).13C NMR (50 MHz, CDC13) 8
170.6, 153.3, 137.7,
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131.6, 128.9, 128.5 (2 C's), 128.4, 127.7, 127.5 (2 C's), 112.8, 109.8, 104.,3
101.4, 70.9, 63.0, 45.0,
20.7. MS (EI, m/z) M+=309. '
Step 4: Compound 82 was prepared in a manner similar manner to compound 69,
using N (2-
acetoxyethyl)-5-benzyloxyindole (2.00 g, 13.5 mmol), oxalyl chloride (1.18
rnL, 13.5 mmol),
acetamide (2.67 g, 40.5 mmol), and a 1M solution of KOtBu (68.0 mL, 68.0
mmol). Standard workup
and purification first by silica gel chromatography, eluting with 2:1
petroleum ether/ethyl acetate,
provided compound 82 as a deep red solid in 62% yield. 1H NMR (200 MHz, DMSO-
d6) 8 10.72 (s,
1H), 8.36 (s, 1H), 7.55-7.31 (m, 7H), 6.98 (dd, J=2.0, 9.OHz, 1H), 6.80 (s,
1H), 5.22 (s, 2H), 4.94 (t,
J=S.IHz, 1H), 4.28 (m, 2H), 3.70 (m, 2H).
Example 83: 3-(N (O-acetoxyethyl)-5-benzyloxyindol-3-yl)-1H pyrrole-2,5-dione
Compound 82 (76 mg, 0.25 mmol), triethylamine (35 q,L, 0.25 mmol), acetic
anhydride (30
~,L, 0.3 mmol), and DMAP (2 mg) were stirred together in THF (5 mL) over
night. Standard aqueous
worlcup and recrystallization from methanol, provided compound 83 in 62%
yield, as a yellow solid.
1H NMR (200 MHz, CDCl3) 8 8.31 (d, J=12.1Hz, 1H), 7.48-7.24 (m, 7H), 7.06 (td,
J=2.0, 8.2Hz, 1H),
6.44 (d, J=12.1Hz, 1H), 5.13 (s, 2H), 4.38 (s, 4H), 1.98 (s, 3H).
Example 84: 3-(N (2-(3,4-dimethoxy)benzoyl)oxyethyl)-5-benzyloxyindol-3-yl)-1H
pyrrole-2,5-
dione
Compound 82 (76 mg, 0.25 mmol), triethylamine (35 ~.L, 0.25 mmol), 3,4-
dimethoxybenzoyl
chloride (60 mg, 0.3 mmol), and DMAP (2 mg) were stirred together in THF (5
mL) over night.
Standard aqueous workup and recrystallization from methanol, provided compound
84 in 75% yield,
as a light yellow solid. 1H NMR (200 MHz, CDC13) 8 10.72 (s, 1H), 8.49 (s,
1H), 7.67 (d, J=9.OHz,
1H), 7.52-7.31 (m, 7H), 7.27 (d, J=l.4Hz, 1H), 6.99 (d, J=8.7Hz, 1H), 6.85 (s,
1H), 5.21 (s, 2H), 4.68
(m, 2H), 4.53 (m, 2H), 3.81 (s, 3H), 3.71 (s, 3H).
Example 85:
Compound 82 (76 mg, 0.25 mmol), triethylamine (35 ~,L, 0.25 mmol), and phenyl
isocyanate
were refluxed together in THF (5 mL) over night. Standard aqueous workup and
recrystallization
from methanol, provided compound 85 in 62% yield, as a yellow solid.
Example 86: Synthesis of 3-(1H 5-methoxyindol-3-yl)-1H pyrrole-2,5-diones
Compound 86 was prepared in a manner similar manner to compound 56, using 5-
methoxyindole (2.00 g, 13.5 mmol), oxalyl chloride (1.18 mL, 13.5 mrnol),
acetamide (2.67 g, 40.5
mmol), and a 1M solution of KOtBu (68.0 rnL, 68.0 mmol). Standard workup and
purification first by
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silica gel chromatography, eluting with 2:1 petroleum ether/ethyl acetate
provided compound 86 as an
light orange solid in 62% yield. 1H NMR (200 MHz, DMSO-d6) 8 11.95 (s, 1H),
10.71 (s, 1H), 8.31
(s, 1H), 7.40 (d, J=8.7Hz, 1H), 7.32 (d, J=2.2Hz, 1H), 6.87 (dd, J=2.2, 8.7Hz,
1H), 6.81 (s, 1H), 3.84
(s, 3H). 13C NMR (50 MHz, acetone-d6) 8 172.7, 172.5, 155.9, 140.1, 131.8,
131.2, 126.7, 114.94,
114.8, 113.2, 113.0, 105.9, 102.4, 55.2. .
Example 87: Synthesis of 3-(N tosyl-5-methoxyindol-3-yl)-1H pyrrole-2,5-dione
Compound 86 (70 mg, 0.289 mmol), triethylamine (48 ~,L, 0.347 mmol), DMAP (10
mg), and
p-toluenesulfonyl chloride (66 mg, 0.347 mmol) were refluxed in THF (5 mL) for
48 hours. Standard
aqueous workup and purification by silica gel chromatography, eluting with 2:1
petroleum ether/ethyl
acetate, provided compound 87 in 43% yield, as a light yellow solid.
Example 88: 3-(N (4-nitrobenzenesulfonyl)-5-methoxyindol-3-yl)-1H pyrrole-2,5-
dione
Compound 86 (44 mg, 0.182 mmol), triethylamine (32 q,L, 0.227 mmol), DMAP (2
mg), and
4-nitrobenzenesulforlyl chloride (50 mg, 0.227 mmol) were refluxed in THF (5
mL) for 48 hours.
Standard aqueous workup and purification by silica gel chromatography, eluting
with 4:1 to 1:1
petroleum ether/ethyl acetate, provided compound 88 in 17% yield, as a light
yellow solid. 1H NMR
(200 MHz, DMSO-d6) 8 11.15 (s, 1H), 8.52 (s, 1H), 8.33 (br s, 4H), 7.91 (dd,
J=1.1, 9.OHz, 1H), 7.39
(s,lH), 7.32 (s, 1H), 7.07 (d, J=9.lHz, 1H), 3.84 (s, 3H). 13C NMR (74.8Hz,
DMSO-d6) 8172.4, 172.2,
157.3, 151.1, 141.1, 136, 129.4, 128.9, 128.7, 128.2, 125.3, 123.6, 115.2,
114.3, 112, 104.2, 55.9.
Example 89: Synthesis of 3-(N allyl-5-methoxyindol-3-yl)-1H pyrrole-2,5-dione
Step 1: 5-Methoxyindole (294 mg, 2.0 mmol) was dissolved in THF (10 mL) and
treated with
1M KOtBu in THF (2.2 mL, 2.2 mmol). After stirring for 1 hour, allyl bromide
(190 ~,L, 2.2 mmol)
was added and the reaction mixture was stirred for an additional 2 hrs.
Standard aqueous workup
provided N Allyl-5-methoxyindole as an off white solid, which was used without
further purification.
1H NMR (200 MHz, CDC13) 8 7.25 (d, J=9.OHz, 1H), 7.13 (d, J=2.SHz, 1H), 7.09
(d, J=2.OHz, 1H),
6.93 (dd, J=2.0, 9.OHz, 1H), 6.51 (d, J=2.SHz, 1H), 6.03 (tdd, J=5.4, 10.4,
19.7Hz, 1H), 5.23 (dd,
J=l.l, 10.4Hz, 1H), 5.14 (dd, J=1.1, 16.9Hz, 1H), 4.90 (d, J=5.4Hz, 2H), 3.91
(s, 3H).
Step 2: Compound 89 was prepared from N Allyl-5-methoxyindole in a similar
fashion as
that described for compound 56, using the crude N Allyl-5-methoxyindole from
above, oxalyl
chloride (192 ~L, 2.2 mmol), acetamide (360 mg, 6.0 mmol), and 1M THF solution
of KOtBu (20
mL, 20.0 mmol). Standard workup and purification using silica gel
chromatography, eluting with 4:1
to 1:1 petroleum ether/ethyl acetate, yielded a light yellow solid in 60%
overall yield. 'H NMR (200
MHz, DMSO-d6) 8 10.75 (s, 1H), 8.34 (s, 1H), 7.45 (d, J=9.OHz, 1H), 7.33 (d,
J=2.OHz, 1H), 6.91
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(dd, J=2.0, 9.OHz, 1H), 6.83 (s, 1H), 6.00 (tdd, J=5.4, 10.4, 19.7Hz, 1H),
5.19 (dd, J=l.l, 10.4Hz,
1H), 5.09 (dd, J=l.l, 16.9Hz, 1H), 4.90 (d, J=5.4Hz, 2H), 3.85 (s, 3H). 13C
NMR (74.8Hz, DMSO-d6)
d 173.4, 173.2, 155.5, 138.9, 133.7, 131.4, 126.9, 120.1, 117.6, 115.0, 112.8,
112.1, 104.5, 102.8,
55.7, 48.6.
Example 90: Synthesis of 3-(N allyl-5-hydroxyindol-3-yl)-1H pyrrole-2,5-dione
Compound 89 (103 mg, 0.366 mmol) and BBr3 (346 mL, 3.66 mmol) were refluxed
together
in CHZCl2 for 16 hours. The resulting solution was treated with 2M HCl (5 mL)
and extracted with
ethyl acetate. A deep blue solid was filtered off, and the organic layer was
washed with water, dried
over anhydrous Mg2S03, filtered, and the solvent removed under reduced
pressure. The resulting
solid was purified by silica gel chromatography, eluting with 2:1 petroleum
ether/ethyl acetate, to
yield compound 90 as a pail yellow solid in 32% yield.
Example 91: Synthesis of 3-(N tosyl-5-hydroxyindol-3-yl)-1H pyrrole-2,5-dione
Compound 87 (15 mg, 0.038 mmol) and BBr3 (7.2 ~,L, 0.076 mmol) were refluxed
together in
CHZCIz for 16 hours. An additional 8 ~,L of BBr3 was added and the reaction
mixture was refluxed
for an additional 24 hours. Standard aqueous/ethyl acetate workup and
purification of the crude solid
by silica gel chromatography, eluting with 2:1 petroleum ether/ethyl acetate,
provided compound 91
as a pail yellow solid in 57% yield. 1H NMR (200Hz, DMSO-d6) 8 11.21 (s, 1H),
9.59 (s, 1H), 8.43 (s,
1H), 8.91 (m, 3H), 8.85 (d, J=7.lHz, 1H), 7.41 (d, J=7.8Hz, 2H), 7.25 (m, 1H),
6.94 (s, 1H), 2.38 (s,
3H). 13C NMR (74.8Hz, DMSO-d6) b 172.3, 172.2, 154.9, 146.0, 136.7, 133.5,
130.4, 129.5, 128.9,
127.5, 126.9, 122.2, 114.9, 114.2, 110.7, 106.1, 21Ø
Example 92: Synthesis of 3-(1H indol-3-yl)-1H pyrrole-2,5-dione
Compound 92 was prepared in a manner similar manner to compound 56, using
indole (4.00
g, 19.7 mmol), oxalyl chloride (1.47 mL, 19.7 mmol), acetamide (1.16 g, 19.7
mmol), and a 1M
solution of KOtBu (59.1 mL, 59.1 mmol). Standard worlcup and purification by
titration with acetone
provided compound 92 as an orange solid 45% yield. 1H NMR (200 MHz, DMSO-d6) 8
12.01 (s,
1H), 10.76 (s, 1H), 8.36 (d, J=7.2Hz, 1H), 7.95 (d, J=6.8Hz, 1H), 7.52 (d,
J=7.OHz, 1H), 7.28-7.15
(m, 2H), 6.77 (s, 1H).
Example 93: Synthesis of 3-(N (p-toluenesulfonyl)indol-3-yl)-1H pyrrole-2,5-
dione
Compound 92 (52 mg, 0.25 mmol), triethylamine (52 ~L, 0.375 mmol), DMAP (10
mg), and
p-toluenesulfonyl chloride (72 mg, 0.375 mmol) were refluxed in THF (5 mL) for
48 hours. Standard
aqueous worlcup and purification by silica gel chromatography, eluting with
4:1 petroleum ether/ethyl
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acetate, provided compound 93 in 66% yield, as a light yellow solid.1H NMR
(200 MHz, DMSO-d6)
8 11.12 (s, 1H), 8.56 (s, 1H), 8.07 (d, J=7.7Hz, 1H), 8.02 (d, J=7.7Hz, 1H),
7.9 (d, J=8.7Hz, 2H), 7.42
(t, J=7.7Hz, 1H), 7.40 (d, J=8.3Hz, 2H), 7.37 (t, J=7.7Hz, 1H), 7.21 (s, 1H),
2.31 (s, 3H). 13C NMR
(74.8Hz, DMSO-d6) S 172.4, 172.3, 146.3, 136.4, 133.9, 133.4, 130.6, 129.1,
127.6, 127.1, 126.0,
124.6, 123.0, 121.5, 113.4, 111.0, 21.1.
Example 94: 3-(N (4-acetamindobenzenesulfonyl)indol-3-yl)-1H pyrrole-2,5-dione
Compound 92 (52 mg, 0.25 mmol), triethylamine (52 ~L, 0.375 mmol), DMAP (10
mg), and
4-acetamindobenzenesulfonyl chloride (88 mg, 0.375 mmol) were refluxed in THF
(5 mL) for 48
hours. Standard aqueous workup and purification by silica gel chromatography,
eluting with 2:1 to
1:1 petroleum ether/ethyl acetate, provided compound 94 in 33% yield, as a
light yellow solid. 1H
NMR (200 MHz, DMSO-d6) b 11.11 (s, 1H), 10.44 (s, 1H), 8.56 (s, 1H), 8.15 (d,
J=6.5Hz, 1H), 8.02
(d, J=9.OHz, 2H), 7.95 (d, J=6.5Hz, 1H), 7.75 (d, J=9.OHz, 2H), 7.46 (t,
J=6.5Hz, 1H), 7.37 (t,
J=6.SHz, 1H), 7.20 (s, 1H), 2.03 (s, 3H). 13C NMR (74.8Hz, DMSO-d6) ~ 172.5,
172.3, 169.3, 145.2,
136.5, 133.8, 129.2, 128.7, 125.9, 124.5, 122.8, 121.9, 121.6, 121.5, 119.0,
113.3, 110.8, 24.1.
Example 95: Synthesis of 3-(N (2-nitrobenzenesulfonyl)indol-3-yl)-1H pyrrole-
2,5-dione
Compound 92 (52 mg, 0.25 mmol), triethylamine (52 ~L, 0.375 mmol), DMAP (10
mg), and
2-nitrobenzenesulfonyl chloride (56 mg, 0.25 mmol) were refluxed in THF (5 mL)
for 48 hours.
Standard aqueous worlcup and purification by silica gel chromatography,
eluting with 2:1 petroleum
ether/ethyl acetate, provided compound 95 in 44% yield, as a light yellow
solid. 1H NMR (200 MHz,
DMSO-d6) 8 11.15 (br s, 1H), 8.71 (s, 1H), 8.65 (s, 1H), 8.53 (d, J=8.2Hz,
2H), 8.07 (dd, J=5.0,
7.6Hz, 1H), 7.91 (t, J=8.OHz, 1H), 7.55-7.36 (m, 2H), 7.24 (s, 1H).
Example 96: Synthesis of 3-(N (4-nitrobenzenesulfonyl)indol-3-yl)-1H pyrrole-
2,5-dione
Compoun..d 92 (52 mg, 0.25 mmol), triethylamine (52 ~L, 0.375 mmol), DMAP (10
mg), and
2-nitrobenzenesulfonyl chloride (56 mg, 0.25 mmol) were refluxed in THF (5 mL)
for 48 hours.
Standard aqueous workup and purification by silica gel chromatography, eluting
with 2:1 petroleum
ether/ethyl acetate, provided compound 96 in 11% yield, as a light yellow
solid. 1H NMR (200 MHz,
DMSO-d6) 8 11.16 (s, 1H), 8.58 (s, 1H), 8.36 (br s, 4H), 8.09 (d, J=7.OHz, 1H)
8.03 (d, J=7.OHz, 1H),
7.51 (t, J=7.OHz, 1H), 7.41 (t, J=7.OHz, 1H), 7.26 (s, 1H).'3C NMR (74.8Hz,
DMSO-d6) 8 172.3,
172.1, 151.1, 141.1, 133.8, 130.9, 128.8, 128.7, 127.6, 126.4, 125.3, 125.0,
123.7, 121.7, 113.3, 111.9.
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Example 97: Synthesis of 3-(N (2-thiophenesulfonyl)indol-3-yl)-1H pyrrole-2,5-
dione
Compound 92 (52 mg, 0.25 mmol), triethylamine (52 ~L, 0.375 mmol), DMAP (10
mg), and
2-thiophenesulfonyl chloride (66 mg, 0.375 mmol) were refluxed in THF (5 mL)
for 48 hours.
Standard aqueous workup and purification by silica gel chromatography, eluting
with 2:1 petroleum
5 ether/ethyl acetate, provided compound 97 in 5 8% yield, as an off white
solid. 1H NMR (200 MHz,
DMSO-d6) 8 11.12 (br s, 1H), 8.47 (s, 1H), 8.04-7.97 (m, 3H), 7.45 (t,
J=7.SHz, 1H), 7.38 (t. J=7.SHz,
1H), 7.19-7.15 (m, 2H). 13C NMR (74.8Hz, DMSO-d6) 8 172.5, 172.3, 137.4,
136.3, 135.7 (2), 133.9,
128.8 (2), 127.7, 126.2, 125.0, 123.2, 121.7, 113.5, 111.6.
10 Example 98: Synthesis of 3-(N octanesulfonylindol-3-yl)-1H pyrrole-2,5-
dione
Compound 92 (52 mg, 0.25 mmol) was dissolved in THF (5 mL) and treated with a
1.0M
THF solution of KOtBu (250 q,L, 0.25 mmol). To the resulting deep red solution
was added neat
butanesulfonyl chloride (36 ~,L, 0.257 mmol) and this mixture was stirred for
16 hours. Standard
aqueous workup and purification by silica gel chromatography, eluting with 3:1
petroleum ether/ethyl
15 acetate, provided compound 98 in 10% yield, as red semi-solid.
Example 99: Synthesis of 3-(1H 5-chloroindol-3-yl)-1H pyrrole-2,5-dione
Compound 99 was prepared in a manner similar manner to compound 56, using 5-
chloroindole (1.00 g, 6.60 mmol), oxalyl chloride (633 ~.L, 7.26 mmol),
acetamide (1.19 g, 19.8
20 mmol), and a 1M solution of KOtBu (38.0 mL, 33.0 mmol). Standard workup and
purification by
tituration with acetone provided compound 99 as a light orange solid in 5%
yield.1H NMR (200
MHz, DMSO-d6) S 12.14 (br s, 1H), 10.78 (br s, 1H), 8.38 (s, 1H), 8.00 (s,
1H), 7.51 (d, J=8.6 Hz,
1H), 7.25 (d, J=1.6 Hz, 1H), 6.89 (s, 1H). MS (EI, m/z) M+=246.
25 Example 100: Synthesis of 3-(1H 6-chloroindol-3-yl)-1H pyrrole-2,5-dione
Compound 100 was prepared in a manner similar manner to compound 56, using 6-
chloroindole (500 mg, 3.30 mmol), oxalyl chloride (317 q,L, 3.62=3 mmol),
acetamide (590 mg, 9.90
mmol), and a 1M solution of KOtBu (16.5 mL, 16.5 mmol). Standard workup and
purification by
tituration with acetone provided compound 100 as a light orange solid in 43%
yield. m.p. 280.2-282.2
30 °C. 1H NMR (200 MHz, DMSO-d6) 8 12.07 (br s, 1H), 10.79 (r s, 1H),
8.37 (d, J=2.4 Hz, 1H), 7.99
(d, J=8.SHz, 1H), 7.56 (s, 1H), 7:18 (d, J=8.5 Hz, 1H), 6.83 (s, 1H). 13C NMR
(57.3 MHz, DMSO-d6)
8 173.2, 172.9, 138.9, 137.1, 131.7, 127.5, 124.3, 121.7, 121.4, 116.2, 112.2,
105.4. MS (EI, m/z)
M+=246.
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Example 101: Synthesis of 3-(1H 7-chloroindol-3-yl)-1H pyrrole-2,5-dione
Compound 101 was prepared in a manner similar manner to compound 56 using 7-
chloroindole (500 mg, 3.30 mmol), oxalyl chloride (317 pL, 3.62 mmol),
acetamide (590 mg, 9.90
minol), and a 1M solution of KOtBu (16.5 mL, 16.5 mmol). Standard workup and
purification by
tituration with acetone provided compound 101 as a light orange solid in 37%
yield. m.p. 247.6-249.8
°C.'H NMR (200 MHz, DMSO-d6) 8 12.39 (br s, 1H), 10.84 (br s, 1H), 8.32
(d, J=3.lHz, 1H), 7.98
(d, J=8.0 Hz, 1H), 7.35 (d, J=7.6Hz, 1H), 7.20 (t, J=7.9Hz, 1H), 6.90 (s, 1H).
13C NMR (57.3 MHz,
DMSO-d6) 8 171.9, 171.7, 137.5, 132.2, 130.2, 130.1, 126.2, 121.3, 121.1
118.2, 115.7, 105.1. MS
(EI, m/z) M+=246.
Example 102: Synthesis of 3-(1H 5-flouroindol-3-yl)-1H pyrrole-2,5-dione
Compound 102 was prepared in a manner similar manner to compound 56, using 5-
fluoroindole (500
mg, 3.70 mmol), oxalyl chloride (355 ~,L, 4.07 mmol), acetamide (655 mg, 11.1
mmol), and a 1M
solution of KOtBu (18.5 mL, 18.5 mmol). Standard workup and purification by
titration with acetone
provided compound 102 as a light orange solid 36% yield. 1H NMR (200 MHz, DMSO-
d6) ~ 12.07
(br s, 1H), 10.74 (br s, 1H), 8.40 (d, J=2.7 Hz, 1H), 7.52 (dd, J=4.8Hz,
8.9Hz, 1H), 7.09 (m, 1H), 6.86
(s, 1H). MS (EI,m/z) M+=230.
Example 103: Synthesis of 3-(1H 6-floixroindol-3-yl)-1H pyrrole-2,5-dione
Compound 103 was prepared in a manner similar manner to compound 56, using 6-
fluoroindole (500 mg, 3.70 mmol), oxalyl chloride (355 ~L, 4.07 mmol),
acetamide (655 mg, 11.1
mmol), and a 1M solution of KOtBu (18.5 mL, 18.5 mmol). Standard workup and
purification by
titration with acetone provided compound 103 as a light orange solid 15%
yield. m.p. 248.5-249.0 °C.
'H NMR (200MHz, DMSO-d6) 8 12.02 (br s, 1H), 10.77 (br s, 1H), 8.33 (d,
J=3.OHz, 1H), 7.96 (dd,
J=8.9Hz, 5,2Hz, 1H), 7.30 (dd, J=10.4Hz, 2.3Hz, 1H), 7.04 (dt, J=7.3Hz, 2.3Hz,
1H), 6.83 (s, 1H).
13C NMR (SOMHz, DMSO-d6) 8 173.1, 172.9, 139.0, 129.5, 121.6, 121.5, 115.8,
109.6, 109.3, 105.4,
98.9, 98.5. MS (EI,m/z) M+=230.
Example 104: Synthesis of 3-(1H 5-nitroindol-3-yl)-1H pyrrole-2,5-dione
Compound 104 was prepared in a manner similar manner to compound 56, using 5-
nitroindole (1.00 mg, 6.17 mmol), oxalyl chloride (565 p,L, 6.48 mmol),
acetamide (1.15 g, 19.4
mmol), and a 1M solution of KOtBu (31.00 mL, 31.0 mmol). Standard workup and
purification by
titration with acetone provided compound 104 as a light orange solid 59 %
yield. IH NMR (200MHz,
DMSO-d6) S 12.56 (br s, 1H), 10.90 (s, 1H), 8.72 (d, J=l.9Hz, 1H), 8.44 (s,
1H), 8.08 (d, J=8.4Hz,
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1H), 7.64 (d, J=8.8Hz, 1H), 6.93 (s, 1H). 13C NMR (SOMHz, DMSO-d6) 8 172.8,
172.6, 139.8, 133.7,
130.3, 124.7, 118.3, 118.1, 116.8, 113.0, 107.0, 92.4.
Example 105: Synthesis of 3-(1-H 5-benzyloxyindol-3-yl)-1H pyrrole-2-one-5-
thione
Compound 105 was prepared in a manner similar manner to compound 56, using 5-
benzyloxyindole (223 mg, 1.0 rnmol), oxalyl chloride (96 pL, 1.1 mmol),
thioacetamide (250 mg, 3.3
mmol), and a 1M solution of KOtBu (5.0 mL, 5.0 mmol). Standard workup and
purification by
tituration with acetone provided compound 105 as a red solid 19% yield.
1H NMR (200 MHz, DMSO-d6) 8 12.07 (s, 1H), 11.93 (s, 1H), 8.38 (d, J=2.8Hz,
1H), 7.53-7.31 (m,
7H), 7.01 (s, 1H), 6.97 (dd, J=2.0, 8.8Hz, 1H), 5.22 (s, 2H). 13C NMR (SOMHz,
DMSO-d6) 8 175.1,
154.5, 137.6, 133.9, 132.2, °131.8, 128.3, 127.7, 127.6, 126.1, 122.0,
113.6, 113.5, 104.9, 104.2, 70Ø
Example 106:
Compound 106 was prepared as a by-product of the reaction described for
Compound 17.
Example 107: 3-hydroxy-4-(indol-3-yl)-pyrrole-2,5-dione
Indole-3-acetamide and dimethyl oxylate were dissolved in THF (10 mL) and
treated with a
1.0M solution of KOtBu (3 equiv). After stirring for 1 hour the reaction
mixture was diluted with
water and the resulting solid filtered, washed with water and dried in vacuo,
to provide a compound
108 as a deep red solid in 65%. 'H NMR (200MHz, acetone-d6) 8 10.67 (br s,
1H), 9.44 (br s, 1H),
8.31 (dd, J=0.8, 8.OHz, 1H), 8.03 (s, 1H), 7.42 (dd, J=0.8, 8.OHz, 1H); 7.16
(dt, J=1.1, 7.3Hz, 1H),
7.08 (dt, J=1.1, 7.3Hz, 1H). 13C NMR (SOMHz, acetone-d6) b 172.2, 169.1,
147.1, 137.3, 127.6,
127.4, 126.8, 123.3, 122.8, 120.4, 112.2. MS (EI, m/z) M+=228.
Example 108: 3-hydroxy-4-(N methylindol-3-yl)-pyrrole-2,5-dione
N Methylindole-3-acetamide and dimethyl oxylate were dissolved in THF (10 mL)
and
treated with a l .OM solution of KOtBu (3 equiv). After stirring for 1 hour
the reaction mixture was
diluted with water and the resulting solid filtered, washed with water and
dried in vacuo, to provide a
compound 108 as a deep red solid in 87%.
1H NMR (200MHz, DMSO-d6) 8 8.90 (br s, 1H), 8.53 (d, 3=8.OHz, 1H), 7.59 (s,
1H), 7.23 (d,
J=8.2Hz, 1H), 7.00 (dt, J=1.2, 6.9Hz, 1H), 6.89 (dt, J=1.2, 6.9Hz, 1H), 3.70
(s, 3H).
Example 109:
Compound 122 was prepared according to the procedure described for compound
56, using 3-
(1-(4-chlorobenzyl)-5-(1-quinolinylmethyl)indolyl)-2,2-dimethylpropianoate (1
equiv), oxalyl
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chloride (1.1 equiv), acetamide (3 equiv) and 1.0M KOtBu in THF (3 equiv).
Standard workup and
purification using silica gel chromatography, eluting with 10:1 methylene
chloride/methanol,
provided compound 109 as an orange solid in 55%. 1H NMR (200MHz, DMSO-d6) 8
10.87 (br s, 1H),
8.39 (d, J=5.6Hz, 1H), 8.06 (d, J=5.6Hz, 1H), 7.98 (d, J=5.6Hz, 1H), 7.79 (d,
J=5.6Hz, 1H), 7.75 (t,
J=5.6Hz, 1H), 7.51 (t, J=5.6Hz, 1H), 7.51 (m, 4H), 6.95 (m, 3H), 6.75 (s, 1H),
5.72 (s, SH), 5.51 (s,
2H), 5.40 (s, 2H).
Example 110:
Compound 84 was refluxing in methylene chloride with 5 equiv of BBr3 for 16
hours.
Aqueous worlcup and purification by silica gel chromatography, eluting with
1:1 acetone/hexane,
provided compound 110 in 54 % yield.
Example 111:
Compound 111 was prepared according to the procedure described for compound
56, using
intermediate H1, oxalyl chloride, acetamide, and 1M KotBu in THF. Purification
by silica gel
chromatography, eluting with 2:1 hexane/acetone, provided compound 111 in 32%
yield.
Example 112: . High/Low Potassium Assay
CGNs were harvested from day 8/9 post-natal CD1 mice, plated on Poly-D-Lycine
coated
plates and incubated for 6 days at 37 °C, with 25 ~tM potassium, under
5% COz. Pretreated cells were
treated with drug 24 hours prior to changing the media to one containing 5 ~,M
potassium and drug.
Cells were assayed 16 hours after the final media change using cell TITER96
(Promega). ICSO was
evaluated as the concentration at which cell death was inhibited by 50%.
Example ICso
K252a 0.30
CEP 1347 - 1
24 10
34 10
42 3
47 3 (100% inhibition at 10
~,M)
48 3
60 10
77 10
95 10
96 10
The compounds formed according to this invention inhibit HK/LK apoptotic cell
death in
CGNs with selected compounds protecting upwards of 100% of the neurons at 10
~m drug
concentrations with ICSO values in the range of 1-10 N,M'. K252a and CEP 1347
displayed ICso values
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of 0.3 and 1 ~,M, respectively. These compounds, however, were toxic at higher
doses, while the
compounds tested herein displayed little or no toxicity in untreated controls.
Example 113: (3-Amyloid Aggregation Assay
CGNs were harvested from day 8/9 post-natal CD1 mice, plated on Poly-D-Lycine
coated
plates and incubated for 6 days at 37 °C under 5% CO2. Pretreated cells
were treated with drug 24
hours prior to A-(3 (25 uM) and drug addition. Cells were assayed 5 days after
A(3 addition using cell
TITER96 (Promega).
Ex- ample ICSO~M)
CEP 1347 toxic X300 nM
2 10
24 10
42 1
64 1
87 10
Linear A(3 rapidly aggregates in CGN cultures, leading to the apoptotic cell
death of
approximately 50% of the neurons after 5 days. Addition of selected compounds
of the formula I
through III saves upwards of 100% of these cells at drug concentrations of 10
~M with ICSO values of
1-10 E.~M. These compounds may be added to the CGN culture 24 hours prior to
linear A[3 addition or
at the time of linear A(3 addition. Similar saves were observed under these
two scenarios. The most
active compounds displayed limited toxicity, less than 5%, in their respective
in vitro controls. As
observed in the HK/LK assays, CEP 1347 displayed limited protection (> 10 % at
concentrations
below 300 nM) and severe toxicity at concentrations greater than 300 nM.
Example 114: Ceramide Killing of CGNs
CGNs were harvested from day 8/9 post-natal CD 1 mice, plated on Poly-D-Lycine
coated
plates and incubated for 6 days at 37 °C under 5% C02. Cells were
pretreated with drug 24 hours prior
to ceramide (100 uM) and drug addition. Cells were assayed 16 hours after
final drug treatments
using cell TITER96 (Promega).
% CGN Survival
at Various
Concentrations
Example 10 ~,M 3 uM 1 .~~M 300 nM 100 nM 30 nM
K252a - - - 0 (250 nM) - -
CEP 1347 - - 1.8 0 (250 nM) - -
24 10.7 - 11.4 - - -
42 18.0 - 20.0 - - -
61 - - 5.1 - 0
64 5.1 - 18.0 - - -
87 9.1 - 0 - - -
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Addition of selected compounds of the formula I through III, 24 hours prior to
the addition of
ceramide, to cultured CGNs provided protection against apoptosis, with 10 to
20% of the cells being
saved at 1-10 ~,M drug concentrations.
Example 115: Glutamate Killing of CGNs
CGNs were harvested from day 8/9 post-natal CD1 mice, plated on Poly-D-Lycine
coated
plates and incubated for 6 days at 37 °C under 5% CO2. Cells were
pretreated with drug 24 hours
prior to glutamate (100 uM) and drug addition. Cells were assayed 16 hours
after final drug treatments
using cell TITER96 (Promega).
% CGN Survival at Various Concentrations
Example 10 pM ~ 1 uM 300 nM 100 nM 30 nM
K252a - - - 4.3 (250 riM) - -
CEP 1347 - - 7.0 50.0 (250 nM) 0 -
24 22.3 - 17.0 - 0 -
64 32.5 - 4.0 - 0
87 - - 11.5 - 0 -
Addition of selected compounds of the formula I through III, either at the
time of glutamate
addition or 24 hours prior to the addition of ceramide, to cultured CGNs
provided protection against
neuronal cell death, with up to 20% of the cells being saved at l-10 ~M drug
concentrations.
Example 116: Cisplatin Killing of CGNs
CGNs were harvested from day 8/9 post-natal CD1 mice, plated on Poly-D-Lycine
coated
plates and incubated for 6 days at 37 °C under 5% COz. The cells were
treated with media containing
drug and cisplatin (25 mg/mL). Cells were assayed 48 hours after final drug
treatments using cell
TITER96 (Promega).
Example ICSO (,~M)
CEP 1347 30 % at 1 ~M
1 10
2 10
3 10
5 10
24 3
34 10
35 0.1
36 0.1
42 3
47 10
48 10
51 1
52 3
64 3
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Addition of selected compounds of the formula I through III, 24 hours prior to
the addition of
cisplatin (25 p,g/mL), to cultured CGNs protected against neuronal apoptosis.
Compounds 35 and 36
displayed ICso of 100 nM (80 % survival at 300 nM) against cisplatin induced
apoptosis in. CGNs.
Several other compounds of the formula I through III displayed ICso values in
the range of 3-10 mM,
with upwards of 80 % neuronal survival. CEP 1347 displayed limited protection
(> 30%) at 1 pM.
Example 117: Cisplatin Filling of Cortical Neurons
Cortical neurons were harvested from day E18 female Sprague-Dawley rats,
plated on Poly-
L-Lycine coated plates and incubated for 14 days at 37 °C under 5% COZ.
The cells pre-treated with
with media containing drug (10 ~M) for 24 hours, followed by cisplatin (35
mg/mL). Cells were
assayed 16-24 hours after final drug treatments using cell TITER96 (Promega).
Exam 1e % Protection at 10 ~M
35 11
36 9
51 23
53 41
CEP 1347 32 at 0.3 ~M
Compound 52 protected cultured cortical neurons protecting 50% of the neurons
at
concentrations of 10 E,tM.
Example 118: NGF Withdrawal Filling of SCG Neurons
SCG neurons were harvested from day 1 post natal Sprague-Dawley rats, plated
on collagen
coated plates and incubated for 5 days in the presence of 50 ng/mL NGF at 37
°C under 5% CO2. The
cells were washed with NGF free media, 4 times at 1 hour intervals, at which
time drug was added.
Cells were assayed 48 hours after final drug treatments using cell TITER96
(Promega).
Exam 1e ICso f uM)
1 20
33 10
42 20
47 10
67 10
97 10
91 20
109 10
Compounds of the formula I through III protect SCG neurons against NGF
withdrawal when
applied at concentration of 10-20 pM.
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Example 119: Etoposide Filling of SHSY-SY
SHSY-SY cells were grown in growth media. Cells are placed at 50,000 cells per
96 well.
Four days latter the cells were treated with drug for 48 hours prior to
etoposide (32 uM) treatment.
On day 6 media was changed to that containing etoposide and drug for 4 hours,
at which point the
media was changed to media containing drug only. Cells are allowed to survive
overnight and then
assessed for viability with metabolic activity measured (WST-1- Beohringer
Mannheime).
CGN Survival at Various Concentrations
Example 10 ~M 3 uM l ~u,M 300 nM 100 nM 30 nM
K252a - - 13.9 - -
CEP 1347 - - 12.5 - -
1 12.5 - 0 - -
69 - - 5.6 - -
70 7.6 - _ _ _
71 26.4 - - - -
73 toxic - - -
99 4.7 - _ _ _
121 - - 5.5 - - -
SHSY-SY cells are members of a neuroblastoma cell line. When SHSY-SYs were
pretreated for 24 hours with selected compounds of the formula I through III,
followed by etoposide,
little or no protection was observed.
Example 120: LANS HAIP 1 Down Regulators
LANS cells were grown in growth media. Cells were plated at cells 50,000 per
well in 96
cells per 96 wells. Five days latter drug was added and the cells were liced
24 hours latter using RLT
buffer. The licate was processed for RNA extraction and RNA levels were
measured on TAQUMEN.
Example Fold Induction
control 1.0
F252a 0.51
56 0.31
57 ° 0.45
58 0.55
59 0.40
60 0.13
61 0.16
62 0.18
63 0.22
70 0.21
87 0.37
Cancer cells became sensitized to apoptosis by the down-regulation of the
IAPs. The use of
small molecules for the down-regulation of the IAPs represents a novel
approach to cancer
chemotherapy.
