Indian Patents
Title of Invention

SUBSTITUTED PYRAZOLOPYRIMIDINES
Abstract The present invention is related to chemical compositions, processes for the preparation thereof and uses of the composition. Particularly, the present invention relates to compositions that include substituted heterobicyclic pyrimidines of Formula (I): wherein R<1>, R<2>, R<3>, R<4>, R<5>, X, W, and ring A are as defined herein; pharmaceutical compositions of substituted heterobicyclic pyrimidines of Formula (I); and their use in the treatment of chronic neurodegenerative diseases, neurotraumatic diseases, depression and/or diabetes. More particularly, the present invention relates to substituted pyrazolopyrimidines of Formula (I).
Full Text SUBSTITUTED PYRAZOLOPYRIMIDINES
FIELD OF THE INVENTION
The present invention is related to chemical compositions, processes for the
preparation thereof and uses of the composition. Particularly, the present invention
relates to compositions that include substituted heterobicyclic pyrimidines of Formula
(I):
wherein R1, R2, R3, R4, R5, X, W, and ring A are as defined herein; pharmaceutical
compositions of substituted heterobicyclic pyrimidines of Formula (I); and their use in
the treatment of chronic neurodegenerative diseases, neurotraumatic diseases,
depression and/or diabetes. More particularly, the present invention relates to
substituted pyrazolopyrimidines of Formula (I).
BACKGROUND OF THE INVENTION
This invention relates to novel substituted heterobicyclic pyrimidine compounds,
in particular substituted pyrazolopyrimidine oxindoles, that act as inhibitors of glycogen
synthase kinase 3 and cyclin dependant kinase 5.
Glycogen synthase kinase 3 (GSK3) is a serine/threonine protein kinase
composed of two isoforms (a and ß encoded by different genes. GSK3 is highly
expressed in the central and peripheral nervous system, with GSK3ß predominating in
the brain. Both isoforms of GSK3 phosphorylate and regulate the activity of several
protein substrates, including glycogen synthase, ß-catenin, pyruvate dehydrogenase,
elongation intiation factor 2b, and tau. GSK3 is regulated by insulin, which stimulates
glycogen synthesis via receptor activation of PI3 kinase and protein kinase B. PKB
phosphorylates GSK3ß on serine 9, resulting in its inactivation. Insulin also activates
protein phosphatase 1. Both of these actions of insulin lead to dephosphorylation and
activation of glycogen synthase (Srivastava and Pandey, Mol Cell Biochem.l82:135-
141,1998; Cohen, Biochem See Trans., 21:555-567,1993), and production of glycogen
from glucose. ß-catenin degradation is increased following phosphorylation by GSK3
(Ikeda, et al., EMBO, 17:1371-1384,1998). The reduction in available ß-catenin may
increase the sensitivity of neurons to amyloid ß (Aß) toxicity (Zhang, et al., Nature,
395:698-702,1998). GSK3ß also phosphorylates pyruvate dehydrogenase and prevents
the conversion of pyruvate to acetyl CoA (Hoshi, et al., PNAS, 93:2719-2723,1996).
This acetyl CoA is critical for the synthesis of acetylcholine, the loss of which is
implicated in the cognitive decline in Alzheimer's Disease (AD). GSK3a regulates
production of Aß from the amyloid precursor protein (Phiel, et al., Nature. 423:435-9,
2003). Two proteases, ß- and ?-secreatase liberate the amino and carboxy terminus
(respectively) of Aß. In a concentration dependant manner, Aß precipitates into toxic,
fibrillary species in the AD brain and is thought to lead to additional sequalae of the
disease. Phosphorylation of eIF2B by GSK-3ß reduces protein translation. eIF2B
activation by IGF1 is mediated by the inactivation of GSK3ß (Welsh, et al., FEBS Letts,
421:125-130,1997). The role of tau phosphorylation by GSK3 will be discussed
following description of CDK5.
Cyclin Dependant Kinase 5 (CDK5) is also a serine/threonine protein kinase,
and is structurally related to GSK 3. CDK5 activation predominates in the nervous
system due to expression of p35, an accessory protein related to cyclins and necessary
for CDK5 activity (Dhavan and Tsai, Nat Rev Mol Cell Biol, 2:749-759,2001). Unlike
CDK1,2,4, and 6 which are active in the cell cycle, CDK5 is activated in neurons after
cell division has ended, following differentiation and expression of p35. CDK5 activity
is regulated by expression of p35 and a calpain-cleaved form of p35, known as p25
(Patzke and Tsai, J Biol Chem, 277:8054-8060,2002). The generation of p25 leads to
increased and mislocalized CDK5 activity since 1) p25 is missing the membrane
localizing portion found in p35, and 2) p25 has a longer resident half life in the
cytoplasm. CDK5 phosphorylates a number of substrates including DARPP-32, NR2a
(NMDA receptor subunit), MEF-2, PSD-95, synaptojanin-1, CRMP2, and tau. DARPP-
32 phosphorylation by CDK5 at thr75 leads to the inhibition of PKA in the dopamine 1
receptor (Dl) signaling cascade, thereby inhibiting Dl signaling (Bibb, et al., Nature,
402:669-671 1997). Facilitation of Dl signaling may be useful for the treatment of
depression or Parkinson's Disease (Chergui, et al., PNAS, 10:2191-2196,2004). NR2a
phosphorylation by CDK5 modulates long term potentiation and may induce apoptotic
cell death following ischemia (Wang, et al., Nat Neurosci., 6:1039-47,2003). CDK5-
dependent phosphorylation of PSD-95 dynamically regulates the clustering of PSD-
95/NMDA receptors at synapses, providing a possible mechanism for rapid changes in
density and/or number of synaptic receptors (Morabito, et. al., J Neurosci., 24:865-876,
2004). CDK5 also phosphorylates the presynaptic phosphatase synaptojanin 1 and
regulates its function both in vitro and in intact synaptosomes (Lee, et. al., PNAS,
101:546-551,2004). CRMP2 is also phosphorylated by CDK5, leading to a reduction in
CRMP-tubulin binding affinity and modulating growth cone collapse. CDK5 primarily
phosphorylates CRMP2 at Ser.522 and GSK3ß secondarily phosphorylates at Thr509.
Dual-phosphorylated CRMP2 s recognized with the antibody 3F4, highly reactive with
the neurofibrillary tangles (NFT) of AD brain (Uchida, et al., Genes Cells, 10:165-179,
2005). Overall, the role of CDK5 in synaptic formation and function is well
substantiated.
Experimental evidence supports a role for both GSK3 and CDK5 in the tangle
and plaque pathology of AD, namely in the tau hyperphosphorylation that leads to NFT
formation. AD brain is characterized by intracellular NFTs and extracellular senile
plaques consisting of Aß deposits. Both of these protein aggregates are thought to
precipitate the neuronal and synaptic loss, leading to the memory loss and cognitive
decline of AD (Hardy, J Mol Ncurosci, 20:203-6, 2003).
NFTs are composed of hyperphosphorylated, aggregated forms of the neuron
specific, cytoskeletal protein tau (Cairns, et. al., J Pathol, 204:438-449,2004). The
primary function of tau is to stabilize neuronal microtubules, to maintain axonal
architecture, and to allow transport of materials both from the cell body to the synapse,
and from the synapse back to the cell body. In AD, tau is hyperphosphorylated at many
serine/threonine residues, leading to poor binding of tau to the microtubule and loss of
trophic interplay between the cell body and the synapse. NFTs represent one of the
characteristic features of the AD brain, and are also present in the brains of individuals
with progressive supranuclear pafcsy, frontotemporal dementia with parkinsonism-17,
Neimann-Pick's disease, corticobasal degeneration, amyotrophic lateral sclerosis,
dementia puglistica, etc.
NFTs are composed of insoluble aggretates of tau protein, hyperphosphorylated
on many serine and threonine residues and formed into paired helical filaments. The
hyperphosphorylation of tau results in a lower affinity for the microtubule and may
represent the first step toward aggregate formation. Both CDK5 and GSK3
phosphorylate tau in both cell-free and cell-based in vitro systems at many of the same
sites present in the AD brain. Antibodies directed against both GSK3 (Pei, et. al., J.
Neuropath. Exp. Neurol., 58: 1010-1019,1999) and CDK5 (Pei, et. al., Brain Res,
797:267-277,1998) decorate the NFTs in the AD brain, demonstrating the close
association between these kinases and the hyperphosphorylated tau that comprises the
tangles. Overexpression of either kinase activity in transgenic animal models (Lucas, et
al., EMBO J., 20:27-39,2001; Cruz, et al., Neuron, 40:471-83,2003) also demonstrates
their ability to hyperphosphorylate tau (both CDK5 and GSK3) and cause the formation
of mature NFTs and neuronal toss (CDK5). Phosphorylation by GSK3 at many epitopes
requires prior phosphorylation by a so called "priming" kinase C-terminal to the GSK3
phosphorylation site (Cohen and Goedert, Nat Rev Drug Discov, 3:479-487,2004).
Interestingly, CDK5 has been implicated as a priming kinase (at phosphoserine 235) for
GSK3, which acts to phosphorylate threonine 231, a site that is phosphorylated early in
the progression of AD NFT pathology (Augustinack, et. al., Acta Neuropathol (Berl).
103:26-35,2002; Li T, Hawkes C, Qureshi HY, Kar S, Paudel HK, Biochemistry,
45:3145-4154,2006).
Other disease states in which GSK3 is thought to play a role include cerebral
ischemia. GSK3 activity is increased in cellular and animal models of both
neurodegeneration and apoptosis, such as cerebral ischemia (Bhat, et al., PNAS,
97:11074-11079,2000). Lithium, as a representative GSK3 inhibitor, is neuroprotective
in these models (Ren, et al., PNAS, USA, 100:6210-6215,2003). Lithium inhibits
GSK3 at concentrations also known to be therapeutic in bipolar disorder (Gould, et al., J
Clin Psychiatry, 65:10-21,2004), implicating GSK3 inhibition as a therapeutic avenue
in this disease.
GSK3 activity is increased in peripheral lymphocytes and brains of patients with
schizophrenia, as evidenced by reduced levels of both the upstream inhibiting kinase
AKT1, and the inhibitory ser9 phosphorylation of GSK3ß (Emamian, et al., Nat Genet,
36:131-137,2004) Clinical treatment leads to normalization of this pathway.
Diabetes mellitus type 2 is characterized by reduced insulin production due to
loss of pancreatic beta cells following a period of reduced insulin sensitivity. With the
insulin receptor signaling dysfunction that is also present in the disease, direct inhibition
of GSK3 has been hypothesized 1o relieve the hyperglycemia and allow for normal
glycogen synthesis and glucose utilization (Wagman, et al., Curr Pharm Des.,10:l 105-
1137,2004).
These compounds as GSK3 inhibitor are indicated to be useful for the treatment
and/or prophylaxis of conditions in which mere is a need for inhibition of GSK3, such
as diabetes, conditions associated with diabetes, chronic neurodegenerative diseases
such as Alzheimer's Disease, Parldnson's Disease, progressive supranuclear palsy,
subacute panencephalitic parkinsonism, postencephalitic parkinsonism, dementia
puglistica, guan-parkinsonial dementia complex, Pick's disease, corticobasal
degeneration, frontotemporal dementia with parkinsonism, Huntington's disease, AIDS
associated dementia, amyotrophic lateral sclerosis, multiple sclerosis, and
neurotraumatic diseases such as acute stroke, mood disorders such as schizophrenia and
bipolar disorders, promotion of functional recovery post stroke, cerebral bleeding
(solitary cerebral amyloid anigopathy), hair loss, obesity, atherosclerotic cardiovascular
disease, hypertension, polycystic ovary syndrome, syndrome X, ischemia, traumatic
brain injury, cancer, leukopenia, Down's syndrome, Lewy body disease, inflammation
and immunodeficiency.
These compounds as CDK5 inhibitors are indicated to be useful for the treatment
and/or prophylaxis of conditions in which there is a need for inhibition of CDK5 such as
the chronic neurodegenerative diseases Alzheimer's disease, Parkinson's Disease,
progressive supranuclear palsy, subacute panencephalitic parkinsonism, postencephalitic
parkinsonism, dementia puglistica, guan-parkinsonial dementia complex, Pick's disease,
corticobasal degeneration, frontotemporal dementia with parkinsonism, Huntington's
disease, AIDS-associated dementia, amyotrophic lateral sclerosis and mood disorders
such as depression.
Thus, there is a need for aovel classes of compounds that possess the beneficial
properties. It has been discovered that a class of compounds, referred to herein as
substituted heterobicyclic pyrimidine compounds, in particular substituted
pyrazolopyrimidine oxindoles, are useful as agents for treating or preventing various
diseases or disorders disclosed herein.
SUMMARY OF THE INVENTION
The present invention in one aspect is directed to various novel compounds of
structure:
wherein R1, R2, R3, R4, R5, X and ring A are as defined herein; and its stereoisomeric
forms, mixtures of stereoisomeric forms, tautomeric forms, or pharmaceutically
acceptable salt forms thereof, wherein the constituent members are defined infra.
Another object of the present invention is to provide pharmaceutical
compositions comprising the compounds of the present invention wherein the
compositions comprise one or more pharmaceutically acceptable excipients and a
therapeutically effective amount of at least one of the compounds of the present
invention, or a pharmaceutically acceptable salt or ester form thereof.
Another object of the present invention is to provide methods of treating or
preventing diseases or disorders, including chronic neurodegenerative diseases is
selected from Alzheimer's Disease, Parkinson's Disease, progressive supranuclear
palsy, subacute panencephalitic parkinsonism, postencephalitic parkinsonism, dementia
puglistica, guan-parkinsonial dementia complex, Pick's disease, corticobasal
degeneration, firontotemporal dementia with parkinsonism, Huntington's disease, AIDS
associated dementia, amyotrophic lateral sclerosis, and multiple sclerosis.
Another object of the present invention is to provide methods of treating or
preventing diseases or disorders, including neurotraumatic disease selected from acute
stroke, mood disorders such as schizophrenia and bipolar disorders, promotion of
functional recovery post stroke, cerebral bleeding (solitary cerebral amyloid
anigopathy), hair loss, obesity, atherosclerotic cardiovascular disease, hypertension,
polycystic ovary syndrome, syndrome X, ischemia, traumatic brain injury, cancer,
leukopenia, Down's syndrome, Lewy body disease, inflammation and
immunodeficiency.
Another object of the present invention is to provide methods of treating
depression.
Another object of the present invention is to provide methods of treating
diabetes.
These and other objects, features and advantages of the substituted
pyrazolopyrimidines will be disclosed in the following detailed description of the patent
disclosure.
DETAILED DESCRIPTION OF THE INVENTION
In a first embodiment, the present invention provides novel compounds of
Formula (I):
and stereoisomeric forms, mixtures of stereoisomeric forms, tautomeric forms, prodrugs,
or pharmaceutically acceptable salt forms thereof, wherein:
W is CH or N;
ring A is
R1, R2, R3, and R4 at each occurrence are independently selected from
H, halo, -OR11, -NO2, -CN, -CF3, -CHF2, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4
haloalkyl, -NR13R14, -NHOR13a, -C(=O)R15, -C(=O)OR15, -OC(=O)R15, -
C(=O)NR13R14, -NR13aC(=O)R15, -NR13aCO2R15, -OC(=O)NR13R14, -
NR13aC(=S)R15, -SR15, -S(=O)R15, -S(=O)2R15, -S(=O)2NR13R14, and C1-C4
alkyl substituted with 0-1 R19;
R5 is H, C1-C6 alkyl or a prodrug of an amino group;
R6 is selected from H;
C1-C6 alkyl substituted by 0-2 R22;
C2-C6 alkenyl substitute.! by 0-2 R22;
C2-C6 alkynyl substituted by 0-2 R22; and
C3-C7 cycloalkyl substituted by 0-3 R22;
R7 is H, -NO2, halo, C1-C4 alkyl or -NR23R24;
X is selected from H, -NR9R10, halo, OR12, -NO2, -CN, -CF3, -CHF2, C1-C4 alkyl,
C2-C4 alkenyl, C2-C4 alkynyl, C1-C4 haloalkyl, -CH2NR9R10, -CH2OR12,
-NHOR16, -C(0)R18, -C(=O)OR18, -OC(O)R18, -C(=O)NR9R10, -
NR16C(=O)R18,
-NR16CO2R18, -OC(=O)I4R9R10, -NR16C(=S)R18, -SR18, -S(=O)R18, -S(=O)2R18,
-S(=O)2NR9R10, and -NE.16S(=O)2R18;
R9 and R10 at each occurrence are; each independently selected from H, -NH2;
C1-C6 alkyl substituted by 0-1 R19;
C2-C6 alkenyl substituted by 0-1 R19;
C2-C6 alkynyl substituted by 0-1 R19;
C6-C10 aryl substituted by 0-5 R19;
C3-C7 carbocyclyl substituted by 0-5 R19; and
5 to 14 membered heterocyclyl group substituted by 0-5 R19, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaiyl group substituted by 0-5 R19, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O, and S;
alternatively, R9 and R10, together with the nitrogen to which they are attached, form a
3-7 membered heterocyclic ring, wherein said 3-7 membered heterocyclic ring
contains a nitrogen atom and optionally a second atom selected from N, O, S,
S(=O) , and S(=O)2, wherein said 3-7 membered heterocyclic ring is substituted
with 0-1 R17;
R11 at each occurrence is independently selected from H, C1-C4 alkyl, and C1-C4
haloalkyl;
R12 at each occurrence is independently selected from H, C1-C4 haloalkyl and C1-C4
alkyl substituted with 0-1 R19;
R13 and R14 , at each occurrence, are independently selected from H, C1-C4 alkyl
substituted with 0-3 R30; and C6-C10 aryl substituted with 0-5 R30;
R 3a at each occurrence is independently selected from H, C1-C4 alkyl, and C6-C10 aryl;
R15 at each occurrence is independently selected from H,
C1-C6 alkyl substituted by 0-1 R30;
C2-C6 alkenyl substituted by 0-1 R30;
C2-C6 alkynyl substituted by 0-1 R30;
C6-C10 aryl substituted by 0-5 R30;
C3-C7 carbocyclyl substituted by 0-5 R30; and
5 to 14 membered heterocyclyl group substituted by 0-5 R30, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R30, wherein said
heteroaryl group comp rises one, two, or three heteroatoms selected from N,
O, and S;
R16 at each occurrence is independently selected from H and C1-C4 alkyl;
R17 is H, -NR23R24, halo, -NO2, -CN, -CF3, C1-C4 haloalkyl, -NHOH, OR25, C(=O)R25,
C(=O)OR25, OC(=O)R25, C(=O)NR23R24, NR23aC(=O)R25, NR23aC02R25,
OC(=O)NR23R24, NR23aC(=-S)R25, SR25, S(=O)R2S, S(=O)2R25; S(=O)2NR23R24,
-NR23aS(=O)2R25, or C1-C4 alkyl substituted by 0-1 R19;
R18 at each occurrence is independently selected from H;
C1-C6 alkyl substituted by 0-1 R30;
C2-C6 alkenyl substituted by 0-1 R30;
C2-C6 alkynyl substituted by 0-1 R30;
C6-C10 aryl substituted by 0-5 R30;
C3-C7 carbocyclyl substituted by 0-5 R30; and
5 to 14 membered heterocyclyl group substituted by 0-5 R30, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R30, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O, and S;
R19 at each occurrence is independently selected from H, -NR23R24, halo, -NO2, -CN, -
CF3, C1-C4 haloalkyl, -NHOH, OR25, C(=O)R25, C(=O)OR25, OC(=O)R25,
C(=O)NR23R24, NR23,C(=O)R25, NR23aCO2R25,OC(=O)NR23R24,
NR23aC(=S)R25, SR25 S(=O)R25, S(=O)2R25; S(=O)2NR23R24, -NR23aS(=O)2R25,
C1-C4 alkyl substituted by 0-1 R30;
C2-C4 alkenyl substituted by 0-1 R30;
C2-C4 alkynyl substituted by 0-1 R30;
C6-C10 aryl substituted by 0-5 R30;
C3-C7 carbocyclyl substituted by 0-5 R30; and
5 to 14 membered hetetocyclyl group substituted by 0-5 R30, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R30, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O, and S;
R22 is H, -NR23R24, -N3, halo, -NO2, -CN, -CF3, CrC4 alkyl, C2-C4 alkenyl, C2-C4
alkynyl, C1-C4 haloalkyl, C3-C7 carbocyclyl, phenyl, -NHOH, OR25, -CH2OR25,
C(=O)R25, C(=O)OR25,OC(=O)R25, C(=O)NR23R24, NR23aC(=O)R25,
NR23aCO2R25, OC(=O)NR23R24, NR23aC(=S)R25, SR25, S(=O)R25, S(=O)2R25;
S(=O)2NR23R24, or -NR23lS(=O)2R25;
R23 and R24 at each occurrence are each independently selected from H or C1-C6 alkyl;
alternatively, R23 and R24, together with the nitrogen to which they are attached, form a
3-7 membered heterocyclic ring, wherein said 3-7 membered heterocyclic ring
contains a nitrogen atom and optionally a second atom selected from N, O and S;
wherein said 3-7 membered heterocyclic ring is substituted with 0-1 C1-C4 alkyl;
R23a at each occurrence is each independently selected from H or C1-C4 alkyl;
R25 at each occurrence is each independently selected from H or C1-C6 alkyl; and
R30 is H, F, Cl, Br, -CF3, C1-C6 alkyl, and C1-C6 alkoxy;
provided when ring A is

then X is -NR9R10.
In a preferred embodiment, W is CH.
In a preferred embodiment, ring A is

In a preferred embodiment, ring A is
In a preferred embodiment, ling A is
In a preferred embodiment, ring A is
In a preferred embodiment, R1, R2, R3, and R4 at each occurrence are
independently selected from H, halo, -OR11, -NO2, -CN, and -CF3.
In a preferred embodiment, Rl is H and R2, R3, and R4 at each occurrence are
independently selected from II, halo, -OR11, -NO2, -CN, and -CF3.
In a preferred embodiment, R1, R3, and R4 are each H and R2 is selected from H,
F, Cl, Br, -OCH3, -NO2, -CN, and -CF3.
In a preferred embodiment, X is H, -NR9R10, halo, C1-C4 alkyl, or OR12.
In a preferred embodiment, X is -NR9R10.
In a preferred embodiment, X is -NHR9.
In a preferred embodiment, R5 is H.
In another first embodiment, the present invention provides novel compounds of
Formula (II);
and stereoisomeric forms, mixtures of stereoisomeric forms, tautomeric forms, prodrugs,
or pharmaceutically acceptable salt forms thereof, wherein:
ring A is
R1, R2, R3, and R4 at each occurrence are independently selected from
H, halo, -OR11, -NO2, -CN, -CF3, -CHF2, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4
haloalkyl, and C1-C4 alkyl;
R5 is H, C1-C6 alkyl or a prodrug of an amino group;
R6 is selected from H;
C1-C6 alkyl substituted by 0-2 R22;
C2-C6 alkenyl substituted by 0-2 R22;
C2-C6 alkynyl substituted by 0-2 R22; and
C3-C7 cycloalkyl substituted by 0-2 R22;
R7 is H, -NO2, halo, C1-C4 alkyl or -NR23R24;
X is H, -NR9R10, halo, OR12, C1-C4 alkyl, or C2-C4 alkenyl;
R9 and R10 at each occurrence are each independently selected from H, -NH2;
C1-C6 alkyl substituted by 0-1 R19;
C2-C6 alkenyl substituted by 0-1 R19;
C2-C6 alkynyl substituted by 0-1 R19;
C6-C10 aryl substituted by 0-5 R19;
C3-C7carbocyclyl substituted by 0-5 R19; and
5 to 14 membered heterocyclyl group substituted by 0-5 R19, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteronuyl group substituted by 0-5 R19, wherein said
heteroaryl group comiprises one, two, or three heteroatoms selected from N,
O,and S;
alternatively, R9 and R10, together with the nitrogen to which they are attached, form a
3-7 membered heterocyclic ring, wherein said 3-7 membered heterocyclic ring
contains a nitrogen atom ;uid optionally a second atom selected from N, O, S,
S(=O), and S(=O)2, wherein said 3-7 membered heterocyclic ring is substituted
with 0-1 R17;
R11 at each occurrence is independently selected from H, C1-C4 alkyl, and C1-C4
haloalkyl;
R12 at each occurrence is independently selected from H, C1-C4 haloalkyl and C1-C4
alkyl substituted with 0-1 R19;
R17 is H or C1-C4 alkyl substituted by 0-1 R19;
R19 at each occurrence is independently selected from H, -NR23R24, halo, -NO2, -CN, -
CF3, C1-C4 haloalkyl, -NHOH, OR25, C(=O)R25, C(=O)OR25, OC(=O)R25,
C(=O)NR23R24, NR23aC(=O)R25, NR23aCO2R25, OC(=O)NR23R24,
NR23aC(=S)R25, SR25, S(=O)R25, S 2R25; S(=O)2NR23R24, -NR23aS(=O)2R25,
C1-C4 alkyl substituted by 0-1 R30;
C2-C4 alkenyl substituted by 0-1 R30;
C2-C4 alkynyl substituted by 0-1 R30;
C6-C10 aryl substituted by C-5 R30;
C3-C7 carbocyclyl substituted by 0-5 R ; and
5 to 14 membered heterocyolyl group substituted by 0-5 R30, wherein said
heterocyclyl group comprises one., two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R30, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O,and S;
R22 is H, -NR23R24, -N3, halo, -NO2, -CN, -CF3, C1-C4 alkyl, C2-C4 alkenyl, C2-C4
alkynyl, C1-C4 haloalkyl, C3-C7 carbocyclyl, phenyl, -NHOH, OR25, -CH2OR25,
C(=O)R25, C(=O)OR25, OC(=O)R25, C(=O)NR23R24, NR23aC(=O)R25,
NR23aC02R25, OC(=O)NR23R24, NR23aC(=S)R25, SR25, S(=O)R25, S(=O)2R25;
S(=O)2NR23R24, or -NR23aS(=O)2R25;
R23 and R24 at each occurrence are each independently selected from H or C1-C6 alkyl;
alternatively, R23 and R24, togetner with the nitrogen to which they are attached, form a
3-7 membered heterocyclic ring, wherein said 3-7 membered heterocyclic ring
contains a nitrogen atom and optionally a second atom selected from N, O and S,
wherein said 3-7 membered heterocyclic ring is substituted with 0-1 C1-C4 alkyl;
R23a at each occurrence is each independently selected from H or C1-C4 alkyl;
R25 at each occurrence is each independently selected from H or C1-C6 alkyl; and
R30 is H, F, Cl, Br, -CF3, C1-C6 alkyl, and C1-C6 alkoxy.
In a preferred embodiment, ring A is
In a preferred embodimen:, ring A is
In a preferred embodiment, ring A is
In preferred embodiment, ring A is
In a preferred embodiment, R1, R2, R3 and R4 at each occurrence are
independently selected from H, F, Cl, Br, -OCH3, -NO2, -CN, and -CF3.
In a preferred embodiment, R1 is H and R2, R3, and R4 at each occurrence are
independently selected from H, F, Cl, Br, -OCH3, -NO2, -CN, and -CF3.
In a preferred embodiment, R1, R3, and R4 are each H and R2 is selected from H,
F, Cl, Br, -OCH3, -NO2, -CN, and -CF3.
In a preferred embodiment, X is -NR9R10.
In a preferred embodiment, X is -NHR9.
In a preferred embodiment, R6 is C1-C6 alkyl substituted by 0-2 R22.
In a preferred embodiment, R6 is C3-C7 cycloalkyl substituted by 0-2 R22.
In a preferred embodiment, R6 is cyclopentyl.
In another first embodiment, the present invention provides novel compounds of
Formula (III):
and stereoisomeric forms, mixtures of stereoisomeric forms, tautomeric forms, prodrugs,
or pharmaceutically acceptable salt forms thereof, wherein:
ring A is
R is selected from
H, halo, -OR11, -NO2, -CN, -CF3, -CHF2, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4
haloalkyl, and C1-C4 alkyl
R5 is H, methyl or a prodrug of an amino group;
R6 is selected from H;
C1-C6 alkyl substituted by 0-2 R22;
C2-C6 alkenyl substituted by 0-2 R22;
C2-C6 alkynyl substituted by 0-2 R22; and
C3-C7 cycloalkyl substituted by 0-2 R22;
X is H, -NR9R10, halo, OR12, C1-C4 alkyl, or C2-C4 alkenyl;
R9 and R10 at each occurrence are each independently selected from H, -NH2;
C1-C6 alkyl substituted oy 0-1 R19;
C2-C6 alkenyl substituted by 0-1 R19;
C2-C6 alkynyl substituted by 0-1 R19;
C6-C10 aryl substituted by 0-5 R19;
C3-C7 carbocyclyl substituted by 0-5 R19; and
5 to 14 membered heterocyclyl group substituted by 0-5 R19, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R19, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O, and S;
alternatively, R9 and R10, together with the nitrogen to which they are attached, form a
3-7 membered heterocyclic ring, wherein said 3-7 membered heterocyclic ring
contains a nitrogen atom and optionally a second atom selected from N, O, S,
S(=O), and S(=O)2, wherein said 3-7 membered heterocyclic ring is substituted
with 0-1 R17;
R11 at each occurrence is independently selected from H, C1-C4 alkyl, and C1-C4
haloalkyl;
R12 at each occurrence is independently selected from H, C1-C4 haloalkyl and C1-C4
alkyl substituted with 0-1 R19;
R17 is H or C1-C4 alkyl substituted by 0-1 R19;
R19 at each occurrence is independently selected from H, -NR23R24, halo, -NO2, -CN, -
CF3, C1-C4 haloalkyl, -NHOH, OR25, C(=O)R25, C(=O)OR25, OC(=O)R25,
C(=O)NR23R24, NHC(=O)R25, NHCO2R25, OC(=O)NR23R24, NHC(=S)R25,
SR25, S(=O)R25, S(=O)2R25; S(=O)2NR23R24, -NHS(=O)2R25,
C1-C4 alkyl substituted by 0-1 R30;
C2-C4 alkenyl substituted by 0-1 R30;
C2-C4 alkynyl substituted by 0-1 R30;
C6-C10 aryl substituted by 0-5 R30;
C3-C7 carbocyclyl substituted by 0-5 R30; and
5 to 14 membered heterocyclyl group substituted by 0-5 R30, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R30, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O, and S;
R22 is H, -NR23R24, -N3, halo, -NO2, -CN, -CF3, C1-C4 alkyl, C2-C4 alkenyl, C2-C4
alkynyl, C1-C4 haloalkyl, C3-C7 carbocyclyl, phenyl, -NHOH, OR25, C(=O)R25,
C(=O)OR25, OC(=O)R25, C(=O)NR23R24, NHC(=O)R25, NHCO2R25,
OC(=O)NR23R24, NHC(=S)R25, SB25, S(=O)R25, S(=O)2R25; S(=O)2NR23R24, or
-NHS(=O)2R25;
R23 and R24 at each occurrence are each independently selected from H or C1-C4 alkyl;
R25 at each occurrence is each independently selected from H or C1-C4 alkyl; and
R30 is H, F, Cl, Br, -CF3, C1-C4 alkyl, and C1-C4 alkoxy.
In a preferred embodiment, R2 is selected from H, F, Cl, Br, -OCH3, -NO2, -CN,
and -CF3.
In a preferred embodiment. R6 is cyclobutyl, cyclopentyl, or cyclohexyl.
In a preferred embodiment, R6 is cyclopentyl.
In a preferred embodiment, X is -NR9R10.
In a preferred embodiment, X is -NHR9.
In a preferred embodiment, R6 is cyclopentyl and X is -NR9R10.
In a preferred embodiment, R2 is selected from F, Cl, Br, -OCH3, -CN, and -CF3;
and X is -NR9R10.
In another first embodiment the present invention provides novel compounds of
Formula (II):
and stereoisomeric forms, mixtures of stereoisomeric forms, tautomeric forms or
pharmaceutically acceptable salt forms thereof, wherein:
ring A is
R1, R2, R3, and R4 at each occurrence are independently selected from H, F, Cl, Br, -
OCH3, -NO2, -CN, and -CF3;
R5 is H;
R6 is selected from H, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, n-pentyl, i-
pentyl, allyl, cyclopentyl, c yclohexyl,
-CH2CH2OCH3, -CH2CH2OCH2CH3, -CH2CH2CH2N3, and -
CH2CH2CH2NHCH3;
R7 is H or -NO2;
X is selected from H, Cl, methyl, ethyl, propyl, butyl,
-OH; -OCH2CH2N(CH3)2; -OCH2CH2(pyrid-3-yl);
-NHCH3; -NCH2CH3; -NHCH(CH3)2; -NHCH2CH2CH2CH3; -
NHCH2CH(CH3)2;
-NHCH2CH2CF3; -NHCH=CH2; -NHCH2CH=CH2;
-NHCH2CH2N(CH3)2; -N(CH3)CH2CH2N(CH3)2; -NHCH2CH2CH2N(CH3)2;
-NHCH2CH2CH2NH(CH3);
-NHCH2CH2NH2; -NHCH2CH2CH2NH2; -N(H)CH2CH(NH2)CH3;
-N(CH3)CH2CH2N(CH2CH3)2; -NHNH2; -NHCH2CH2NHC(=O)CH3;
-N(CH2CH2OCH3)2; -N(H)CH2CH2OCH3; -N(H)CH2CH2CH2OCH3;
-N(H)CH2CH2OCH2CH3;-N(H)CH2CH2OCH2CH2CH3;
In another first embodiment, the present invention provides novel compounds of
Formula (I) selected from the following Examples:
and pharmaceuticaliy acceptable salt forms thereof.
In a second embodiment, the present invention provides a method for treatment
of diseases comprising administering to a subject in need thereof a therapeutically
effective amount of a compound of Formula (I), or a pharmaceuticaliy acceptable salt
thereof, wherein the disease is selected from a chronic neurodegenerative disease, a
neurotraumatic disease, depression and diabetes.
In a preferred embodiment, the present invention provides a method of treating
or preventing chronic neurodegenerative diseases selected from Alzheimer's Disease,
Parkinson's Disease, progressiva supranuclear palsy, subacute panencephalitic
parkinsonism, postencephalitic parkinsonism, dementia puglistica, guan-parkinsonial
dementia complex, Pick's disease, corticobasal degeneration, frontotemporal dementia
with parkinsonism, Huntington's disease, AIDS associated dementia, amyotrophic
lateral sclerosis, and multiple sclerosis.
In a more preferred second embodiment the present invention provides a method
wherein the compound is administered for the treatment of Alzheimer's Disease (AD).
In a third embodiment, the present invention provides a pharmaceutical
composition comprising a compound of Formula (I), or a pharmaceuticaliy acceptable
salt or ester form thereof, and one or more pharmaceuticaliy acceptable excipients.
In a preferred third embodiment, the present invention provides a pharmaceutical
composition comprising a therapeutically effective amount of compound of Formula (I),
or a pharmaceuticaliy acceptable salt or ester form thereof, and one or more
pharmaceuticaliy acceptable exepients.
In a fourth embodiment, the present invention provides for the use of compounds
of formula (I) or pharmaceuticaliy acceptable salts thereof for the manufacture of a
medicament for the treatment of a disease or disorder, as disclosed herein.
These and other objects, features and advantages of the substituted
pyrazolopyrimidines will be disclosed in the following detailed description of the patent
disclosure.
Definitions
The following terms and expressions contained herein are defined as follows:
As used herein, the term "about" refers to a range of values from ± 10% of a
specified value. For example, the phrase "about 50 mg" includes ± 10% of 50, or from
45 to 55 mg.
As used herein, a range of values in the form "x-y" or "x to y", or "x through
y", include integers x, y, and the integers there between. For example, the phrases "1-
6", or "1 to 6" or "1 through 6" are intended to include the integers 1,2, 3,4,5, and 6.
Preferred embodiments include each individual integer in the range, as well as any
subcombination of integers. For example, preferred integers for "1-6" can include 1,2,
3, 4, 5, 6,1-2,1-3,1-4,1-5, 2-3, 2-4, 2-5, or 2-6, etc.
As used herein "stable compound" or "stable structure" refers to a compound
that is sufficiently robust to survive isolation to a useful degree of purity from a reaction
mixture, and preferably capable of formulation into an efficacious therapeutic agent.
The present invention is directed only to stable compounds.
As used herein, the term "alkyl" refers to a straight-chain, or branched, alkyl
group having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, sec-butyl, tert-butyl, pentyl, isoamyl, neopentyl, 1-ethylpropyl, 3-
methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, hexyl, etc. The alkyl moiety of
alkyl-containing groups, such as alkoxy, alkoxycarbonyl, and alkylaminocarbonyl
groups, has the same meaning as alkyl defined above. Lower alkyl groups, which are
preferred, are alkyl groups as defined above which contain 1 to 4 carbons, such as
methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl. A designation
such as "C1-C4 alkyl" refers to an alkyl radical containing from 1 to 4 carbon atoms.
As used herein, the term "alkenyl" refers to a straight-chain, or branched,
hydrocarbon group of 2 to 6 carbon atoms having at least one carbon-carbon double
bond. A designation "C2-C6 alkenyl" refers to an alkenyl radical containing from 2 to 6
carbon atoms. Examples of alkenyl groups include, but are not limited to, ethenyl,
propenyl, isopropenyl, butenyl, pentenyl, 2,4-pentadienyl, etc. Preferred alkenyl groups
include ethenyl and propenyl.
As used herein, the term "alkynyl" refers to a straight chain, or branched
hydrocarbon chains of 2 to 6 carbon atoms having at least one carbon-carbon triple
bond. A designation "C2-C6 alkynyl" refers to an alkynyl radical containing from 2 to 6
carbon atoms. Examples include, but are not limited to, ethynyl, propynyl, isopropynyl,
3,5-hexadiynyl, etc.
As used herein, the term "alkylene" refers to a substituted or unsubstituted,
branched or straight chained hydrocarbon of 1 to 6 carbon atoms, which is formed by
the removal of two hydrogen atoms. A designation such as "C1-C4 alkylene" refers to
an alkylene radical containing from 1 to 4 carbon atoms. Examples include, but are not
limited to, methylene (-CH2-), ethylene (-CH2CH2-), ethylidene (-CH(CH3)-), propylene
(-CH2CH2CH2-), iso-propylene (-CH(CH3)CH2-), propylidene (-CH(CH2CH3)-),
butylene (-CH2CH2CH2CH2-), etc.
As used herein, the term "cycloalkylene" refers to a saturated or partially
saturated mono- or bicyclic alkyl ring system containing 3 to 10 carbon atoms, which is
formed by the removal of two hydrogen atoms. A designation such as "C3-C6
cycloalkylene" refers to a cycloalkyl radical containing from 3 to 6 ring carbon atoms.
Preferred cycloalkylene groups include those containing 3,4, 5, or 6 ring carbon atoms.
Examples of cycloalkylene groups include such groups as cyclopropylene (-C3H4-),
cyclobutylene (-C4H6-), cyclopentylene (-C5H8-), cyclopentenylene (-C5H6-),
cyclohexylene (-C6H10-), and cyclohexenylene (-C6H8-).
As used herein, the term "phenylene" refers to a phenyl group with an
additional hydrogen atom removed, i.e. a moiety with the structure of (-C6H4-).
As used herein, the terms "carbocycle", "carbocyclic" or "carbocyclyl" refer
to a substituted or unsubstituted, stable monocyclic or bicyclic hydrocarbon ring system
which is saturated, partially saturated or unsaturated, and contains from 3 to 10 ring
carbon atoms. Accordingly the carbocyclic group may be aromatic or non-aromatic,
and includes the cycloalkyl and aryl compounds defined herein. The bonds connecting
the endocyclic carbon atoms of a carbocyclic group may be single, double, triple, or part
of a fused aromatic moiety.
As used herein, the term "cycloalkyl" refers to a saturated or partially saturated
mono- or bicyclic alkyl ring system containing 3 to 10 carbon atoms. A designation
such as "C3-C7 cycloalkyl" refers to a cycloalkyl radical containing from 3 to 7 ring
carbon atoms. Preferred cycloalkyl groups include those containing 3,4, 5, or 6 ring
carbon atoms. Examples of cycle alkyl groups include such groups as cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, pinenyl, and adamantanyl.
As used herein, the term "cycloalkenyl" refers to partially unsaturated mono- or
bicyclic alkenyl ring system containing 5 to 10 carbon atoms. A designation such as
"C5-C10 cycloalkenyl" refers to a cycloalkenyl radical containing from 5 to 10 ring
carbon atoms and one or more double bonds. Preferred cycloalkenyl groups include
those containing 5 or 7 ring carton atoms. Examples of cycloalkenyl groups include
such groups as cyclopentenyl, cyclohexenyl, and cycloheptenyl.
As used herein, the term "aryl" refers to a substituted or unsubstituted, mono- or
bicyclic hydrocarbon aromatic r ng system having 6 to 10 ring carbon atoms. Examples
include phenyl and naphthyl. Preferred aryl groups include unsubstituted or substituted
phenyl and naphthyl groups. Included within the definition of "aryl" are fused ring
systems, including, for example, ring systems in which an aromatic ring is fused to a
cycloalkyl ring. Examples of such fused ring systems include, for example, indane,
indene, and tetrahydronaphthalere.
As used herein, the term "arylene" refers to an aryl group with an additional
hydrogen atom removed, i.e. an aryl group bonded through two carbon atoms, for
example phenylene.
As used herein, the term "heteroarylene" refers to a heteroaryl group with an
additional hydrogen atom removed, i.e. a heteroaryl group bonded through two carbon
atoms, for example furan-2,5-diyl; or a heteroaryl group bonded through a carbon atom
and a nitrogen atom, for example pyrrol-l,2-diyl.
As used herein, the term "laeterocycloalkylene" refers to a heterocycloalkyl
group with an additional hydroger. atom removed, i.e. a heterocycloalkyl group bonded
through two carbon atoms or a heterocycloalkyl group bonded through a carbon atom
and a nitrogen atom.
As used herein, the terms "heterocycle", "heterocyclic" or "heterocyclyl" refer
to a substituted or unsubstituted carbocyclic group in which the ring portion includes at
least one heteroatom such as O, N, or S. The nitrogen and sulfur heteroatoms may be
optionally oxidized, and the nitrogen may be optionally substituted in non-aromatic
rings. Heterocycles are intended to include heteroaryl and heterocycloalkyl groups.
Examples of heterocyclic groups include pyrrolyl, furanyl, thienyl, pyrazolyl,
imidazolyl, thiazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxathiolyl, oxadiazolyl,
triazolyl, oxatriazolyl, furazanyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl,
pyridazinyl, triazinyl, indolyl, isoindolyl, indazolyl, benzofuranyl, isobenzofuranyl,
purinyl, quinazolinyl, quinolyl, isoquinolyl, benzoimidazolyl, benzothiazolyl,
benzothiophenyl, thianaphthenyl, benzoxazolyl, benzisoxazolyl, cinnolinyl,
phthalazinyl, naphthyridinyl, and quinoxalinyl, as well as, pyrrolidinyl, pyrrolinyl,
imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, pyrazalinyl, piperidyl,
piperazinyl, morpholinyl, thiomorpholinyl, tetrahydromranyl, dithiolyl, oxathiolyl,
dioxazolyl, oxathiazolyl, pyranyl, oxazinyl, oxathiazinyl, and oxadiazinyl.
As used herein, the term "heterocycloalkyl" refers to a 3 to 7 membered
cycloalkyl group in which one or more ring carbon atoms are replaced by at least one
hetero atom such as -O-, -N-, or -S-. Examples of heterocycloalkyl groups include
pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl,
pyrazalinyl, piperidyl, piper azinyl, morpholinyl, thiomorpholinyl, tetrahydrofuranyl,
dithiolyl, oxathiolyl, dioxazolyl, oxathiazolyl, pyranyl, oxazinyl, oxathiazinyl, and
oxadiazinyl.
As used herein, the tt:rm "heteroaryl" refers to an aromatic group containing 5
to 14 ring carbon atoms in which one or more ring carbon atoms are replaced by at least
one hetero atom such as -O-, -N-, -S-, or -Se-. Examples of heteroaryl groups include
pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, isoxazolyl,
oxazolyl, oxathiolyl, oxadiazolyl, triazolyl, oxatriazolyl, furazanyl, tetrazolyl, pyridyl,
pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, thiadiazolyl, picolinyl, indolyl, isoindolyl,
indazolyl, benzofuranyl, isobenzofuranyl, purinyl, quinazolinyl, quinolyl, isoquinolyl,
benzoimidazolyl, benzothiazolyl, benzothiophenyl, thianaphthenyl, benzoxazolyl,
benzisoxazolyl, cinnolinyl, phthalazinyl, naphthyridinyl, and quinoxalinyl. Included
within the definition of "heteroaryl" are fused ring systems, including, for example, ring
systems in which an aromatic ring is fused to a heterocycloalkyl ring. Examples of such
fused ring systems include, for example, phthalamide, phthalic anhydride, indoline,
isoindoline, tetrahydroisoquinoline, chroman, isochroman, chromene, and isochromene.
As used herein, "halo" or "halogen" includes fluoro, chloro, bromo, and iodo.
As used herein, the term "haloalkyl" refers to an alkyl group having one or
more halogen substituents. Example haloalkyl groups include CF3, C2F5, CHF2, CCl3,
CHC12, C2Cl5, and the like. An alkyl group in which all of the hydrogen atoms are
replaced with halogen atoms can be referred to as "perhaloalkyl." Examples
perhaloalkyl groups include CF3 and C2F3.
As used herein, the term "subject" or "mammal" refers to a warm blooded
animal such as a mammal, preferably a human, or a human child, which is afflicted
with, or has the potential to be afflicted with, one or more diseases and conditions
described herein.
As used herein, a "therapeutically effective amount" refers to an amount of a
compound of the present invention effective to prevent or treat the symptoms of
particular disorder. Such disorders include, but are not limited to, those pathological
and neurological disorders associated with the aberrant activity of the receptors
described herein, wherein the treatment or prevention comprises inhibiting, inducing, or
enhancing the activity thereof by contacting the receptor with a compound of the present
invention.
As used herein, the term "pharmaceutically acceptable" refers to those
compounds, materials, compositions, and/or dosage forms which are, within the scope
of sound medical judgment, suitable for contact with the tissues of human beings and
animals without excessive toxicity, irritation, allergic response, or other problem
complications commensurate vith a reasonable benefit/risk ratio.
As used herein, the term "unit dose" refers to a single dose which is capable of
being administered to a patient, and which can be readily handled and packaged,
remaining as a physically and chemically stable unit dose comprising either the active
compound itself, or as a pharmaceutically acceptable composition, as described
hereinafter.
All other terms used in the description of the present invention have their
meanings as is well known in the art.
In another aspect, the present invention is directed to pharmaceutically
acceptable salts of the compounds described above. As used herein, "pharmaceutically
acceptable salts" includes salts of compounds of the present invention derived from the
combination of such compounds with non-toxic acid or base addition salts.
Acid addition salts include inorganic acids such as hydrochloric, hydrobromic,
hydroiodic, sulfuric, nitric and phosphoric acid, as well as organic acids such as acetic,
citric, propionic, tartaric, glutamic, salicylic, oxalic, methanesulfonic, para-
toluenesulfonic, succinic, and benzoic acid, and related inorganic and organic acids.
Base addition salts include those derived from inorganic bases such as
ammonium and alkali and alkaline earth metal hydroxides, carbonates, bicarbonates, and
the like, as well as salts derived from basic organic amines such as aliphatic and
aromatic amines, aliphatic diamines, hydroxy alkamines, and the like. Such bases
useful in preparing the salts of this invention thus include ammonium hydroxide,
potassium carbonate, sodium bicarbonate, calcium hydroxide, methylamine,
diethylamine, ethylenediamine, cyclohexylamine, ethanolamine and the like.
In addition to pharmaceutically-acceptable salts, other salts are included in the
invention. They may serve as intermediates in the purification of the compounds, in the
preparation of other salts, or in the identification and characterization of the compounds
or intermediates.
The pharmaceutically acceptable salts of compounds of the present invention can
also exist as various solvates, such as with water, methanol, ethanol,
dimethylformamide, ethyl acetate and the like. Mixtures of such solvates can also be
prepared. The source of such solvate can be from the solvent of crystallization, inherent
in the solvent of preparation or crystallization, or adventitious to such solvent. Such
solvates are within the scope of the present invention.
The present invention also encompasses the pharmaceutically acceptable
prodrugs of the compounds disclosed herein. As used herein, "prodrug" is intended to
include any compounds which are converted by metabolic processes within the body of
a subject to an active agent that has a formula within the scope of the present invention.
Since prodrugs are known to enhance numerous desirable qualities of pharmaceuticals
(e.g., solubility, bioavailability, manufacturing, etc.) the compounds of the present
invention may be delivered in prodrug form. Conventional procedures for the selection
and preparation of suitable prodrug derivatives are described, for example, in Prodrugs,
Sloane, K. B., Ed.; Marcel Dekker: New York, 1992, incorporated by reference herein
in its entirety. Accordingly, prodrugs include, for example, compounds of the present
invention wherein a hydroxy, amino, or carboxy group is bonded to any group that,
when the prodrug is administered to a mammalian subject, cleaves to form a free
hydroxyl, free amino, or carboa ylic acid, respectively. Examples include, but are not
limited to, acetate, formate and benzoate derivatives of alcohol and amine functional
groups; and alkyl, carbocyclic, aryl, and alkylaryl esters such as methyl, ethyl, propyl,
iso-propyl, butyl, isobutyl, sec-butyl, tert-butyl, cyclopropyl, phenyl, benzyl, and
phenethyl esters, and the like.
As used herein, "prodrug of an amino group" is intended to include a chemical
moiety bonded to an amino group on a compound of the present invention, wherein
when the compound of the present invention is administered to a mammalian subject,
the chemical moiety bonded to :he amino group cleaves to form a free amino,
respectively. Examples include, but are not limited to, acetate, formate and benzoate
derivatives of amine functional groups, as well as alkyl-C(=O)-, alkenyl-C(=O)-,
alkynyl-C(=O)-, carbocyclyl-C(=O)-, carbocyclylalkyl-C(=O)-, alkyl-S(=O)2-,
carbocyclyl-S(=O)2-, carbocyclylalkyl-S(=O)2-, alkyl-NHC(=O)-, carbocyclyl-
NHC(=O)-, carbocyclylalkyl-NHC(=O)-, alkyl-OC(=O)-, carbocyclyl-OC(=O)-,
carbocyclylalkyl-OC(=O)-, alkyl-NH-C(=O)-NHS(=O)2-, carbocyclyl-NH-C(=O)-
NHS(=O)2-, alkyl-S(=O)2-NH-C(=O)-, and carbocyclyl-S(=O)2-NH-C(=O)-groups.
It is recognized that compounds of the present invention may exist in various
stereoisomeric forms. As such, the compounds of the present invention include both
diastereomers and enantiomers. The compounds are normally prepared as racemates
and can conveniently be used as such, but individual enantiomers can be isolated or
synthesized by conventional techniques if so desired. Such racemates and individual
enantiomers and mixtures thereof form part of the present invention.
It is well known in the art how to prepare and isolate such optically active forms.
Specific stereoisomers can be prepared by stereospecific synthesis using
enantiomerically pure or enantiomerically enriched starting materials. The specific
stereoisomers of either starting materials or products can be resolved and recovered by
techniques known in the art, such as resolution of racemic forms, normal, reverse-phase,
and chiral chromatography, recrystallization, enzymatic resolution, or fractional
recrystallization of addition salts formed by reagents used for that purpose. Useful
methods of resolving and recovering specific stereoisomers described in Eliel, E. L.;
Wilen, S.H. Stereochemistry of Organic Compounds; Wiley: New York, 1994, and
Jacques, J, et al. Enantiomen, Racemates, and Resolutions; Wiley: New York, 1981,
each incorporated by reference herein in their entireties.
It is further recognized that functional groups present on the compounds of
Formula (I) may contain protecting groups. For example, the amino acid side chain
substituents of the compounds of Formula (I) can be substituted with protecting groups
such as benzyloxycarbonyl or t-butoxycarbonyl groups. Protecting groups are known
per se as chemical functional groups mat can be selectively appended to and removed
from functionalities, such as hydroxyl groups and carboxyl groups. These groups are
present in a chemical compound to render such functionality inert to chemical reaction
conditions to which the compound is exposed. Any of a variety of protecting groups
may be employed with the present invention. Preferred protecting groups include the
benzyloxycarbonyl (Cbz; Z) group and the tert-butyloxycarbonyl (Boc) group. Other
preferred protecting groups according to the invention may be found in Greene, T.W.
and Wuts, P.G.M., "Protective Groups in Organic Synthesis" 2d. Ed., Wiley & Sons,
1991.
Synthesis
The compounds of the present invention may be prepared in a number of
methods well known to those skilled in the art, including, but not limited to those
described below, or through modifications of these methods by applying standard
techniques known to those skilled in the art of organic synthesis. All processes
disclosed in association with the present invention are contemplated to be practiced on
any scale, including milligram, gram, multigram, kilogram, multikilogram or
commercial industrial scale.
It will be appreciated that he compounds of the present invention may contain
one or more asymmetrically substituted carbon atoms, and may be isolated in optically
active or racemic forms. Thus, all chiral, diastereomeric, racemic forms and all
geometric isomeric forms of a structure are intended, unless the specific stereochemistry
or isomeric form is specifically indicated. It is well known in the art how to prepare
such optically active forms. For example, mixtures of stereoisomers may be separated
by standard techniques including, but not limited to, resolution of racemic forms,
normal, reverse-phase, and chiral chromatography, preferential salt formation,
recrystallization, and the like, or by chiral synthesis either from active starting materials
or by deliberate chiral synthesis of target centers.
As will be readily understood, functional groups present on the compounds of
Formula (I) may contain protecting groups. For example, the amino acid side chain
substituents of the compounds of Formula (I) can be substituted with protecting groups
such as benzyloxycarbonyl or t-butoxycarbonyl groups. Protecting groups are known
per se as chemical functional groups that can be selectively appended to and removed
from functionalities, such as hydroxyl groups and carboxyl groups. These groups are
present in a chemical compound to render such functionality inert to chemical reaction
conditions to which the compound is exposed. Any of a variety of protecting groups
may be employed with the present invention. Preferred protecting groups include the
benzyloxycarbonyl (Cbz; Z) group and the tert-butyloxycarbonyl (Boc) group. Other
preferred protecting groups according to the invention may be found in Greene, T.W.
and Wuts, P.G.M., "Protective Groups in Organic Synthesis" 2d. Ed., Wiley & Sons,
1991.
General routes to prepare the Examples of the present invention are shown in the
Schemes and examples that follow. The reagents and starting materials are
commercially available and/or, using well-known techniques, can be readily synthesized
by one of ordinary skill in the art. All substituents in the synthetic Schemes, unless
otherwise indicated, are as previously defined.
Compounds of invention can be synthesized following various synthetic
schemes disclosed herein.
Patent Experimental
The following Examples are exemplary only, and are not intended to limit the invention.
Compound 2 and 3
3-Amino-1-cyclopentyl-1H-pyrazole-4-carbonitrile
5-Amino-1-cyclopentyl-lH-pyrazole-4-carbonitrile
3-Amino-4-pyrazolecarbonitrile 1 (Acros, 4.32g, 40.0 mmol), cyclopentylbromide
(Acros, 7.15g, 48 mmol) and anhydrous potassium carbonate (Fisher, 6.60g, 48 mmol)
were suspended in 30 mL anhydrous DMF and heated at 80°C under argon overnight.
An additional 3.5g (23.5 mmol) of cyclopentylbromide and 3.3g (24 mmol) of
potassium carbonate were added and the reaction was subjected to an additional six
hours at 80°C. The reaction was permitted to cool and the DMF was removed on a
rotary evaporator. Water was added (100 mL) and the organics were extracted with
dichloromethane (3 X 100 mL). The combined dichloromethane fractions were washed
with water (50 mL) and brine (50 mL) and were dried (magnesium sulfate).
Concentration of the organics afforded a solid which was subjected to flash
chromatography on silica gel (2:1 hexane: ethyl acetate). Two white solids were
obtained: 3 (1.67g, 24%) elutes first and 2 (4.56g, 65%) elutes second. Compound 2:
mp 129-131°C; MS (ES+calculated: 176.22; found: 177.05 M+H). HPLC (100% purity,
Pure compound 2 may be obtained without chromatography in 44% yield by trituration
of the crude solid with a minimum amount of dichloromethane. Compound 2 is
relatively insoluble in dichloromethane whereas 3 dissolves easily.
Compound 3
5-Amino-1-cyclopentyl-1H-pyrazole-4-carbonitrile
Alternate Synthesis of 3: Cyclopentylhydrazine hydrochloride1 (1.08g, 10 mmol) was
dissolved in 100 mL anhydrous ethanol. Sodium methoxide (540 mg, 10 mmol) was
added in one portion and the reaction mixture was stirred for ten minutes.
Ethoxymethylenemalononitrile (Acros, 1.22 g, 10 mmol) was then added in small
portions over several minutes. The reaction mixture was heated at 70°C under argon
overnight. The reaction mixture was concentrated and subjected to flash
chromatography on silica gel (stepwise elution: dichloromethane followed by 1:1
hexane: ethyl acetate) to afford 600 mg (34%) of compound 3 - identical in all respects
with the material obtained above.
Compound 5 and 6
1-Allyl-3-amino-1H-pyrazole-4-carbonitrile
1 -Allyl-5-amino-1H-pyrazole-4-carbonitrile
3-Amino-4-pyrazolecarbonitrile 1 (Acros, 1.08g, 10.0 mmol), allylbromide (Acros,
1.45g, 12 mmol) and anhydrous potassium carbonate (Fisher, 1.65g, 12 mmol) were
suspended in 10 mL anhydrous DMF and heated at 80°C under argon overnight. The
solution was concentrated. Water was added (100 mL) and the organics were extracted
with dichloromethane (3 X 100 mL). The combined dichloromethane fractions were
washed with water (50 mL) and brine (50 mL) and were dried (magnesium sulfate).
Concentration of the organics afforded a solid which was subjected to flash
chromatography on silica gel (gradient elution 2:1 to 3:2 hexane: ethyl acetate). 589 mg
(40%) of a white solid was obtained which was seen by NMR to contain an inseparable
mixture of 5 and 6 in a 2.1.1 ratio. This mixture was used without further purification.
Compound 7 and 8
3-Amino-l-cyclohexyl-1H-pyrazole-4-carbonitrile
5-Amino-l-cyclohexyl-1H-pyrazole-4-carbonitrile
3-Amino-4-pyrazolecarbonitrile 1 (Acros, 6.48g, 60.0 mmol), cyclohexylbromide
(Acros, 11.74g, 72 mmol), and anhydrous potassium carbonate (Fisher, 9.9g, 72 mmol
were combined in 40 mL of DMF and heated at 80°C under argon overnight. The
reaction was permitted to cool to room temperature. Water was added (100 mL) and the
organics were extracted with dichloromethane (3 X 100 mL). The combined
dichloromethane fractions were washed with water (50 mL) and brine (50 mL) and were
dried (magnesium sulfate). Concentration of the organics afforded a solid which was
subjected to flash chromatography on silica gel (gradient elution 3:1 to 2:1 hexane: ethyl
acetate). Separation of the isomers was not fully achieved but a few fractions containing
pure product were combined to afford 7 - a white solid (1.18g, 10%). Compound 7:
mp 169-171°C; HPLC (100% purity, retention time 8.416 minutes - Method B); 1H
NMR (400 MHz, DMSO-d6) d 8 08 (s, 1H), 5.48 (br s, 2H), 3.87 (m, 1H), 1.92 (m, 2H),
1.75 (m, 2H), 1.61 (m, 2H), 1.31 (m, 2H), 1.16 (m, 2H).
Alternate Synthesis of Compound 8: Cyclohexanone (Acros, 19.6g, 200 mmol) and
tert-butylcarbazate (Acros, 26.4g, 200 mmol) were combined in 350 mL dry hexane and
stirred under argon for 54 hour. The mixture was then subjected to reflux temperature
for 1.5 hours and permitted to cool to room temperature. A white solid formed on
cooling which was removed by filtration and dried in vacuo - 40.31 g (95%): mp 147-
149°C ; MS (ES+calculated: 212.29; found: 235.05 M+Na). 1H NMR (400 MHz,
t-Butylcarboxycyclohexanone hydrazone generated above (40.02g, 189 mmol) was
dissolved in a mixture of 175 mL tetrahydrofuran and 225 mL anhydrous methanol.
Sodium cyanoborohydride (Acros, 14.3g, 227 mmol) was added in portions over ten
minutes and the mixture was stirred under argon overnight at room temperature. 135
mL of 6N hydrochloric acid were then added dropwise and the mixture was refluxed for
one hour. The reaction was permitted to cool to room temperature and a white solid was
removed by filtration. The mother liquor was concentrated and residual water was
removed by azeotroping with ethanol on a rotary evaporator (3 X 100 mL). The mixture
was concentrated to dryness and was taken up into hot isopropanol (~300 mL). The
solid that was present was removed by filtration and the mother liquor was concentrated
to ½ volume at which point an equal volume of ethyl ether was added. This caused
cyclohexylhydrazine hydrochloride to precipitate as a white solid. Yield (20.0g, 93%).
To cyclohexylhydrazine hydrochloride (7.52g, 50 mmol) generated above in 500 mL
absolute ethanol was added sodium methoxide (Aldrich, 2.7g, 50 mmol). The mixture
was stirred briefly and ethoxymethylmalononitrile (Acros, 6.11 g, 50 mmol) was added
in small portions over twenty minutes. The reaction mixture was then heated at 70°C
under argon overnight. On cooling, the reaction was concentrated and subjected to flash
chromatography on silica gel (1:1 hexane: ethyl acetate) affording 6.3g (66%) of a light
brown solid. Compound 8: mp 99-103°C; MS (ES+calculated: 190.25; found: 191.15
M+H). HPLC (97% purity, retention time 11.135 minutes - Method A); 1H NMR (400
MHz, DMSO-d6) d 7.50 (s, 1H), 6.49 (br s, 2H), 4.02 (m, 1H), 1.80-1.10 (m, 10H).
Compound 10 and 11
3-Amino-l-cyclohexyl-1H-pyrazole-4-carboxylic acid ethyl ester
5-Amino-l-cyclohexyl-1H-pyrazole-4-carboxylic acid ethyl ester
Ethyl-3-Aminopyrazole-4-carboxylate 9 (Acros, 15.5g, 100.0 mmol),
cyclohexylbromide (Acros, 21.9 g, 130 mmol), anhydrous potassium carbonate (Fisher,
27.6g, 200 mmol), Adogen 464 (Acros, 2.5g) and aqueous sodium hydroxide (0.1 mL of
a 12.5M solution) were combined in 250 mL of toluene and refluxed under argon
overnight. Additional cyclohexylbromide (21.9g, 130 mmol) and potassium carbonate
(27.6g, 200 mmol) were then added and the reaction mixture was resubjected to the
reaction conditions for an additional 24 hours. The reaction was permitted to cool to
room temperature and the organics were washed with 100 mL water. The organic layer
was separated and dried (magnesium sulfate). Concentration of the organics afforded a
solid which was subjected to flash chromatography on silica gel (gradient elution 9:1 to
6:1 to 3:1 hexane: ethyl acetate) to afford two principle products. Compound 11 (923
mg, 4%) elutes first and Compound 10 (1.755g, 7%) elutes second. A considerable
amount of material was present in mixed fractions. Compound 10: MS (ES calculated:
237.30; found: 238.10 M+H). HPLC (98% purity, retention time 13.623 minutes -
Method A); 1H NMR (400 MHz, DMSO-d6) d 7.44 (s, 1H), 6.19 (br s, 2H), 4.43 (q,
J=7Hz, 2H), 4.05 (m, 1H), 1.83-1.22 (m, 10H), 1.23 (t, J=7Hz, 3H). Compound 11:
MS (ES+calculated: 237.30; found: 238.14 M+H). HPLC (100% purity, retention time
13.408 minutes - Method A); 1H NMR (400 MHz, DMSO-d6) d 7.87 (s, 1H), 5.29 (br
s, 2H), 4.15 (q, J=7Hz, 2H), 3.89 (m, 1H), 1.97-1.25 (m, 10H), 1.24 (t, J=7Hz, 3H).
Compound 12
3-Amino-1-cyclopertyl-1H-pyrazole-4-carboxylic acid amide
To concentrated sulfuric acid (Fisher, 8 mL) at 0°C was added 2 (4.24g, 24.0 mmol) in
small portions. The reaction was permitted to warm to room temperature and was
stirred for two hours. At the end of this period all solid had dissolved. This viscous
mixture was then added slowly (violent) to 100 mL concentrated ammonium hydroxide
solution (Fisher). The mixture was stirred for ten minutes and the white solid mat
formed was collected by filtration, was washed with water, and was dried in vacuo.
Yield: 3.838 g (82%). Compound 12: mp 179-181°C; MS (ES+calculated: 194.24;
found: 195.12 M+H). HPLC (100% purity, retention time 5.225 minutes - Method B);
Compound 13
l-Allyl-3-amino-1H-pyrazole-4-carboxylic acid amide
l-Allyl-5-amino-1H-pyrazole-4-carboxylic acid amide
The mixture of solids derived from the preparation of 5 and 6 (589 mg, 4.0 mmol) was
treated with concentrated sulfuric acid (1 mL) as for the preparation of 12 above.
Following neutralization with concentrated ammonium hydroxide (10 mL) and filtration
a mass of white solid was obtained which shrank considerably when washed with water.
The resulting product was dried in vacuo affording 282 mg (42%) of 13 as a white solid.
Compound 14 was determined to be present in the water wash but was not isolated from
this reaction. Compound 13: mp 100-101°C; MS (ES+calculated: 166.18; found:
167.12 M+H). HPLC (94% puriiy, retention time 5.141 minutes - Method A); 1H NMR
(400 MHz, DMSO-d6) d 7.87 (s, 1H), 7.19 (br s, 1H), 6.69 (br s, 1H), 5.95 (m, 1H),
5.34 (br s, 2H), 5.22 (d, J=lHz, 1H), 5.22 (m, 1H), 5.17 (m, 1H), 4.48 (d, J=6Hz, 2H).
Compound 15
5-Arnino-l-cyclohexyl-1H-pyrazole-4-carboxylic acid amide
Compound 8 (6.3g, 33.2 mmol) was treated with concentrated sulfuric acid (12 mL) as
for the preparation of 12 above. The product was isolated via neutralization with
concentrated ammonium hydroxide (225 mL), washing with water, and drying in vacuo
as described for 12. 4.04g (58%) of a white solid, Compound 15 were obtained.
Compound 15: mp 274-278°C; MS (ES+calculated: 208.27; found: 209.14 M+H).
HPLC (97% purity, retention time 6.498 minutes - Method A); 1H NMR (400 MHz,
DMSO-d6) d 7.61 (s, 1H), 7.08 (or s, 1H), 6.58 (br s, 1H), 6.14 (br s, 2H), 3.97 (m, 1H),
1.86-1.58 (m, 7H), 1.44-1.28 (m, 2H), 1.28-1.06 (m, 1H).
Compound 16
5-Amino-1-cyclopentyl-1H-pyrazole-4-carboxylic acid amide
Compound 3 (3.34g, 19.0 mmol) was treated with concentrated sulfuric acid (6 mL) as
for the preparation of 12 above. The product was isolated via neutralization with
concentrated ammonium hydrox de (80 mL), washing with water, and drying in vacuo
as described for 12. 3.64g (99%) of a fluffy white powder, Compound 16, was
obtained. Compound 16: MS [ES+calculated: 194.24; found: 195.10 M+H). HPLC
(97%) purity, retention time 1.729 minutes; - Method C); 1H NMR (400 MHz, DMSO-
d6) d 7.62 (s, 1H), 7.10 (br s, 1H), 6.60 (br s, 1H), 6.14 (br s, 2H), 4.52 (m, 1H), 1.97-
1.67 (m, 6H), 1.63-1.50 (m, 2H).
Compound 17
2-Cyclopentyl-2,5 dihydro-pyrazolo[3,4-d]pyrimidin-4-one
Compound 12 (3.87g, 20 mmol) was suspended in 60 mL triethylorthoformate (Acros)
and refluxed under argon overnight (~150°C). The reaction was concentrated and the
solid obtained was triturated in ether, collected by filtration, and dried in vacuo to afford
3.782g (93%) of a white solid. Compound 17: mp 271-273°C; MS (ES+calculated:
204.23; found: 205.06 M+H). HPLC (100%) purity, retention time 6.104 minutes -
Method B); 1H NMR (400 MHz, DMSO-d6) d 11.65 (br s, 1H), 8.55 (s, 1H), 7.91 (s,
1H), 4.86 (m, 1H), 2.12 (m, 2H), 2.00 (m, 2H), 1.83 (m, 2H), 1.66 (m, 2H). Compound
17 may also be prepared from 12 by refluxing with 2 equivalents of formamidine acetate
in methoxyethanol. Concentration of the solution at the end of the reaction and
neutralizing with ammonium hydroxide solution generates 17 identical in all respects
with that derived from the triethylorthoformate procedure.
Compound 18
2-Cyclohexyl-2,5dihydro-pyrazolo[3,4-d]pyrimidin-4-one
Compound 10 (1.8g, 7.6 mmol) was combined with formamidine acetate (Acros, 1.58g,
15.2 mmol) in 50 mL methoxyethanol (Acros) and refluxed under argon overnight.
Starting material was still evident. Additional formamidine acetate (700 mg, 6.7 mmol)
was added and the mixture was refluxed an additional 24 hours. The reaction was
concentrated and the solid was treated with 100 mL 0.1N ammonium hydroxide
solution. The product was then isolated by filtration, was washed with water, and was
dried in vacuo to afford 1.03g (62%) of a tan solid. Compound 18: mp 287-289°C; MS
(ES calculated: 218.26; found: 2 .7.54 M-H). HPLC (97%) purity, retention time 7.959
minutes - Method A); 1H NMR (400 MHz, DMSO-d6) d 11.64 (br s, 1H), 8.53 (s, 1H),
7.90 (s, 1H), 4.30 (m, 1H), 2.03 (m, 2H), 1.86-1.58 (m, 5H), 1.39 (m, 2H), 1.22 (m, 1H).
Compound 19
2-Allyl-2,5-dihydro-pyrazolo[3,4-d]pyrimidin-4-one
Compound 13 (282 mg, 1.70 mmol) was refluxed under argon in 5 mL
triethylorthoformate overnight. The reaction was concentrated and the crude product
was triturated with cold ethanol. Filtration afforded a white solid which was dried in
Compound 20
1-Cyclopentyl-1,5 -dihydro-pyrazolo[3,4-d]pyrimidin-4-one
In a similar fashion as for the preparation of 19, compound 16 (3.639g, 18.8 mmol) was
refluxed in 75 mL triethylorthoformate affording 3.1 llg (81%) of 20 as white crystals.
Compound 20: mp 235-237°C; MS (ES+calculated: 204.23; found: 205.19 M+H).
HPLC (100%) purity, retention lime 2.674 minutes - Method C); 1H NMR (400 MHz,
DMSO-d6) d 12.12 (br s, 1H), 8.05 (s, 1H), 5.13 (m, 1H), 2.07 (m, 2H), 1.96 (m, 2H),
1.86 (m,2H), 1.67 (m, 2H).
Compound 21
1-Cyclohexyl-1,5- dihydro-pyrazolo[3,4-d]pyrimidin-4-one
In a similar fashion as for the preparation of 18, compound 15 (4.04g, 19.4mmol) was
reacted with formamidine acetate: (4.03g, 38.8 mmol) in 100 mL methoxyethanol to
afford 3.717g (88%) of 21 as a van solid. Compound 21: mp 255-257°C; MS (ES~
calculated: 218.26; found: 217.15 M-H). HPLC (98%) purity, retention time 7.564
minutes - Method B); 1H NMR (400 MHz, DMSO-d6) d 12.13 (br s, 1H), 8.14 (s, 1H),
4.56 (m, 1H), 1.98-1.58 (m, 6H), 1 69 (m, 1H), 1.42 (m, 2H), 1.22 (m, 1H).
Compound 22
4-Chloro-2-cyclopenty]-2H-pyrazolo[3,4-d]pyrimidine
N,N-Dimethylaniline (Acros, 1(1 mL) was added to Compound 17 (3.10g, 13.9 mmol)
dissolved in phosphorus oxychloride (Acros, 90 mL) and the mixture was refluxed
under argon at 110°C for 90 minutes. Excess phosphorus oxychloride was removed in
vacuo and the dark syrup was poured into ice water. The organics were extracted with
three 50 mL portions of ether. The ether extracts were combined, were washed with
water and brine, and were dried (magnesium sulfate). Concentration of the ether
afforded a dark oil which was purified by flash chromatography on silica gel (gradient
elution: 1-3% methanol: dichloromethane) to afford 2.74g (88%) of a green oil.
Compound 22: MS (ES+calculaled: 222.68; found: 223.12 M+H). HPLC (85%) purity,
retention time 9.882 minutes - Method B); 1H NMR (400 MHz, CDC13) d 8.83 (s, 1H),
8.17 (s, 1H), 4.99 (m, 1H), 2.28 (m, 2H), 2.19 (m, 2H), 2.00 (m, 2H), 1.78 (m, 2H).
Compound 23
4-Chloro-2-cyclohexyl-2H-pvrazolo[3,4-d]pyrimidine
In a similar fashion as for the preparation of Compound 22, Compound 23 (0.50g, 2.29
mmol) was treated with 25 mL phosphorus oxychloride and 3 mL N,N-dimethylaniline.
Following flash chromatography on silica gel (gradient elution: 1-3% methanol:
dichloromethane) there was obtained 360mg (66%) of a yellow-green oil. Compound
23: MS (ES+calculated: 236.71; Found: 237.24 M+H). HPLC (94%) purity, retention
time 10.416 minutes - Method B); 1H NMR (400 MHz, DMSO-d6) d 8.97 (s, 1H), 8.79
(s, 1H), 4.59 (m, 1H), 2.12 (m, 2H), 1.93-1.86 (m, 4H), 1.70 (m, 1H), 1.46 (m, 2H), 1.27
(m, 1H).
Compound 24
4-Chloro-2-cyclohexyl-2H-pyrazolo[3,4-d]pyrimidine
In a similar fashion as for the preparation of Compound 22, Compound 19 (110mg, 0.63
mmol) was treated with 5 mL phosphorus oxychloride and 0.5 mL N,N-dimethylaniline.
Following flash chromatography on silica gel (gradient elution: 1-3-5% methanol:
dichloromethane) 77mg (63%) of a dark yellow oil was obtained. Compound 24: MS
(ES+calculated: 194.62; found: 95.07 M+H). 1H NMR (400 MHz, DMSO-d6) d 8.50
(s, 1H), 7.92 (s, 1H), 6.04 (m, 1H), 5.33-5.11 (m, 2H), 4.92 (m, 2H).
Compound 25
4-Chloro-1-eyclopentyl-1H-pyrazolo[3,4-d]pyrimidine
In a similar fashion as for the preparation of compound 22, Compound 20 (1.5g, 7.35
mmol) was treated with 75 mL pnosphorus oxychloride and 6 mL N,N-dimethylaniline.
The reaction was subjected to an ether work-up as for 22 above (without columning) to
afford 1.734g (quantitative) of a yellow solid which was used without further
purification. Compound 25: MS (ES+calculated: 222.68; found: 223.19 M+H).
Compound 26
4-Chloro-1-cyclohexyl-1H-pyrazolo[3,4-d]pyrimidine
In a similar fashion as for the preparation of Compound 22, Compound 21 (1.5g, 6.88
mmol) was treated with 70 mL phosphorus oxychloride and 6 mL N,N-dimethylaniline.
The reaction was subjected to an ether work-up as for 22 above (without columning) to
afford 800mg (of a dark yellow oil which was used without further purification.
Compound 26: MS (ES+calculated: 236.71; found: 237.18 M+H).
Example 29
3-[9-((1R,2S,3R,4R)-2,3-Dihydroxy-4-methoxymethyl-cyclopentyl)-9H-purin-6-yl]-2-
oxo-2,3-dihydro-1H-indole-5-carbonitrile
To 5-cyanooxindole (Combiblocks, 50.6mg, 0.32 mmol) and N,N,N',N'-
tetramethylethylenediamine (Acros, 0.10 mL, 0.64 mmol) in 10 mL anhydrous THF
under argon at -78°C was added lithium diisopropylamine (Acros, 0.32 mL of a 2.0M
solution in THF/heptane, 0.64 mmol). The solution was stirred for fifteen minutes at
which point a solution of Compound 27 (104mg, 0.307 mmol)2 in 10 mL THF was
added dropwise at such a rate as to maintain the temperature below -50°C. The reaction
was stirred for ten minutes, external cooling was removed, and the reaction was
permitted to warm to room temperature. An oil bath was applied and the mixture was
refluxed overnight. The reaction was concentrated, water (50 mL) was added and the
organics were extracted into dichloromethane (3 X 50 mL). The dichloromethane
extracts were combined, were washed with water (50 mL) and brine (50 mL) and were
dried (magnesium sulfate). Removal of the dichloromethane followed by flash
chromatography on silica gel (gradient elution: 1-3% methanol: dichloromethane)
afforded ~60 mg of an acetonice [MS (ES+calculated: 460.50; found: 461.24 M+H)].
which was treated immediately with a mixture of trifluoroacetic acid (24 mL) and water
(3 mL) for 30 minutes at room temperature. The reaction mixture was concentrated and
partitioned between saturated sodium bicarbonate solution (50 mL) and
dichloromethane (50 mL). A solid formed at the interface which was filtered off. [The
dichloromethane layer was determined to contain only polar impurities.] The material
isolated at the interface was wished with water and was dried in vacuo to afford 15 mg
(12%) of the diol as a yellow solid. Example 29: mp 230-236°C (dec); MS
(ES+calculated: 420.43: found: 421.21 M+H). HPLC (95%) purity, retention time 8.089
Example 30
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
To 5-cyanooxindole (Combiblocks, 158.2mg, 1.00 mmol) in 10 mL anhydrous
tetrahydrofuran under argon was added N,N,N',N'-tetramethylethylenediamine (Acros,
0.30 mL, 2.00 mmol). The solution was cooled to -78°C and lithium diisopropylamide
(Acros, 1.0 mL of a 2.0M solution in THF/hexane, 2.00 mmol) was added dropwise.
The reaction was stirred for fifteen minutes at which point a solution of Compound 22
(236mg, 1.06 mmol) in 10 mL anhydrous tetrahydrofuran was added dropwise. The
reaction was stirred an additional fifteen minutes and was warmed to room temperature
for ½ hour. The mixture was then refluxed overnight. The reaction was quenched by
addition of a small amount of a saturated ammonium chloride solution and concentrated.
Dichloromethane (50 mL) and water (50 mL) were added and undissolved solid was
filtered off. The solid was washed with dichloromethane and was taken up into a small
amount of N,N-dimethylformamide and concentrated onto a small amount of silica gel.
The silica containing product was applied to the top of a silica gel column and flash
chromatography was effected (gradient elution: 1-3-5-10% methanol: dichloromethane
to 1% ammonium hydroxide: 10% methanol: 89% dichloromethane) to afford 124 mg
(36%) of a yellow-orange solid. Example 30: mp >300°C (dec); MS (ES+calculated:
344.28: found: 345.17 M+H). HPLC (100%) purity, retention time 12.154 minutes -
J=9 Hz), 7.42 (m),7.07 (d, J=8 Hz), 6.92 (m), 5.22 (m), 4.97 (m), 3.25 (m), 2.25-1.53
(m, 8H).
Example 31
5-Bromo-3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-1,3-dihydro-
indol-2-one
In a similar fashion as for the preparation of 30 above, 5-bromooxindole (Cornbiblocks,
99.6 mg, 0.47 mmol) was reacted with Compound 22 (110 mg, 0.5 mmol). Upon
completion, the reaction was concentrated and the crude product obtained was purified
by flash chromatography on silica gel (gradient elution: 1-3-5-10% methanol:
dichloromethane) to afford 143 mg (76%) of a yellow solid. Example 31: mp 319-
322°C (dec); MS (ES+calculated: 398.27; found: 398.44, 399.81 M+H). HPLC (100%)
Example 32
6-Chloro-3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-1,3-dihydro-
indol-2-one
In a similar fashion as for the preparation of 31 above, 6-chlorooxindole (Cornbiblocks,
78.7mg, 0.47 mmol) was reacted with Compound 22 (110 mg, 0.5 mmol). The crude
product was purified by flash chromatography on silica gel (gradient elution: 1-3-5%
methanol: dichloromethane) to afford 67 mg (40%) of a rust colored solid. Example 32:
Example 33
3-(2-Cyclopentyl-2H-pyrazclo[3,4-d]pyrimidin-4-yl)-5-nitro-1,3-dihydro-indol-2-one
In a similar fashion as for the preparation of 31 above, 5-nitrooxindole (Combiblocks,
50 mg, 0.28 mmol) was reacted with Compound 22 (63 mg, 0.284 mmol). The crude
product was purified by flash chromatography on silica gel (gradient elution: 1-3-5%
methanol: dichloromethane) to afford 30 mg (29%) of a rust colored solid. Example 33:
MS (ES- calculated: 364.37; fcund: 364.03 M+). HPLC (76%) purity, retention time
Example 34
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-trifluoromethyl-l,3-dihydro-
indol-2-one
In a similar fashion as for the preparation of 31 above, 5-trifluoromethyloxindole
(Combiblocks, 55.3 mg, 0.275 mmol) was reacted with Compound 22 (63 mg, 0.284
mmol). The crude product was purified by flash chromatography on silica gel (gradient
elution: 1-3-5% methanol: dichloromethane) to afford 90 mg (84%) of a golden solid.
Example 34: mp 292-293°C; MS (ES+calculated: 387.37; found: 388.18 M+H). HPLC
Example 35
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5,7-difluoro-l,3-dihydro-indol-2-
one
In a similar fashion as for the preparation of 31 above, 5,7-difluoromethyloxindole
(Combiblocks, 47 mg, 0.28 mmol) was reacted with Compound 22 (65 mg, 0.293
mmol). The crude product was purified by flash chromatography on silica gel (gradient
elution: 1-3-5% methanol: dichloromethane) to afford 19 mg (19%) of a yellow solid.
Example 35: mp 296-297°C; MS (ES+calculated: 355.35; found: 386.16 M+H). HPLC
Example 36
5-Chloro-3-(2-Cyclopentyl-2H- pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-1,3-dihydro-
indol-2-one
In a similar fashion as for the preparation of 31 above, 5-chlorooxindole (Combiblocks,
47 mg, 0.28 mmol) was reacted with Compound 22 (65 mg, 0.293 mmol). The crude
product was purified by flash chromatography on silica gel (gradient elution: 1-3-5%
methanol: dichloromethane) to afford 73 mg (74%) of a yellow solid. Example 36: mp
290-292°C; MS (ES+calculated: 353.81; found: 354.26 M+H). HPLC (80%) purity,
Example 37
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indo-2-one
In a similar fashion as for the preparation of 31 above, 5-hydroxyindole (Combiblocks,
45 mg, 0.338 mmol) was reacted with Compound 22 (75 mg, 0.338 mmol). The crude
product was purified by flash chromatography on silica gel (gradient elution: 1-3-5%
methanol: dichloromethane) to afford 70 :mg (65%) of a yellow-orange solid. Example
37: mp 289-292°C; MS (ES+cilculated: 319.37; found: 320.18 M+H). HPLC (87%)
Example 38
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yI)-2-oxo-2,3-dihydro-1H-indole-6-
carbonitrile
In a similar fashion as for the preparation of 31 above, 6-cyanooxindole (Combiblocks,
80 mg, 0.506 mmol) was reacted with Compound 22 (111 mg, 0.5 mmol). The crude
product was purified by flash chromatography on silica gel (gradient elution: 1-3-5-
10% methanol: dichloromethane) to afford 49 mg (28%) of a yellow solid. Example 38:
mp 340-344°C; MS (ES+calculated: 344.38; found: 345.17 M+H). HPLC (98%) purity,
Example 39
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-7-
carbonitrile
In a similar fashion as for the preparation of 31 above, 7-cyanooxindole (Combiblocks,
80 mg, 0.506 mmol) was reacted with Compound 22 (111 mg, 0.5 mmol). The crude
product was purified by flash chromatography on silica gel (gradient elution: 1-3-5%
methanol: dichloromethane) to afford 31 mg (18%) of a yellow solid. Example 39: mp
>380°C; MS (ES+calculated: 344.38; found: 345.16 M+H). HPLC (100%) purity,
Example 40
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-fluoro-l,3-dihydro-indol-2-one
In a similar fashion as for the preparation of 31 above, 5-fluorooxindole (Combiblocks,
76 mg, 0.50 mmol) was reacted with Compound 22 (111 mg, 0.5 mmol). The crude
product was purified by flash cltromatography on silica gel (gradient elution: 1-3-5%
methanol: dichloromethane) to afford 82 mg (49%) of a yellow solid. Example 40: mp
256-260°C; MS (ES+calculated: 337.36; found: 338.18 M+H). HPLC (99%) purity,
Example 41
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-6-fluoro-l,3-dihydro-indol-2-one
In a similar fashion as for the preparation of 31 above, 6-fluorooxindole (Combiblocks,
76 mg, 0.50 mmol) was reacted with Compound 22 (111 mg, 0.5 mmol). The crude
product was purified by flash chromatography on silica gel (gradient elution: 1-3-5%
methanol: dichloromethane) to afford 29 mg (17%) of a yellow solid. Example 41: mp
258-263°C; MS (ES+calculated: 337.36; found: 338.15 M+H). HPLC (96%) purity,
Example 42
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-4,5-difluoro-l,3-dihydro-indol-2-
one
In a similar fashion as for the preparation of 31 above, 4,5-difluorooxindole
(Combiblocks, 85mg, 0.50 mmol) was reacted with Compound 22 (111 mg, 0.5 mmol).
The crude product was purified by flash chromatography on silica gel (gradient elution:
1-3-5% methanol: dichlorome thane) to afford 94 mg (53%) of a yellow solid. Example
42: mp 280-284°C; MS (ES+ calculated: 355.35; found: 356.18 M+H). HPLC (99%)
Example 43
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5,7-dinitro-1,3-dihydro-indol-2-
one
In a similar fashion as for the preparation of 31 above, 5,7-dinirrooxindole
(Combiblocks, 112 mg, 0.506 mmol) was reacted with Compound 22 (111 mg, 0.5
mmol). The crude product was purified by flash chromatography on silica gel (gradient
elution: 1-3-5% methanol: the dichloromethane) to afford 41 mg (20%) of a yellow solid.
Example 43: mp >300°C; MS. (ES+calculated: 409.36; found: 410.19 M+H). HPLC
Example 44
l-(2-Cyclopentyl-2H-pyrazoIo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-benzoimidazol-2-one
In a similar fashion as for the preparation of 31 above, 2-hydroxybenzimidazole (Acros,
68 mg, 0.506 mmol) was reacted with Compound 22 (111 mg, 0.5 mmol). The crude
product was purified by flash chromatography on silica gel (gradient elution: 1-3%
methanol: dichloromethane) to afford 47 mg (29%) of a yellow solid. Example 44: mp
236-240°C: MS (ES+calculatt,d: 320.36: found: 321.17 M+H). HPLC (95%) purity.
Example 45
3-(2-Cyclohexyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
To 5-cyanooxindole (Combibbcks, 34mg, 0.21 mmol) in 10 mL anhydrous
tetrahydrofuran under argon was added N,N,N',N'-tetramethylethylenediamine (Acros,
0.06 mL, 0.40 mmol). The solution was cooled to -78°C and lithium diisopropylamide
(Acros, 0.21 mL of a 2.0M solution in THF/hexane, 0.42 mmol) was added dropwise.
The reaction was stirred for fifteen minutes at which point a solution of Compound 23
(50mg, 0.21 mmol) in 10 mL anhydrous tetrahydrofuran was added dropwise. The
reaction was stirred an additional fifteen minutes and was warmed to room temperature
for 0.5 hour. The mixture was then refiuxed overnight. The reaction was concentrated
and subjected to flash chromatography on silica gel (gradient elution: 1-3-5% methanol:
dichloromethane to afford 60mg (80%) of a yellow solid. Example 45: mp >325°C;
Example 46
5-Bromo-3-(2-cyclohexyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-l,3-dihydro-indol-2-one
In a similar fashion as for the preparation of 45 above, 5-bromooxindole (Combiblocks,
45mg, 0.21 mmol) was reacted with Compound 23 (50mg, 0.21 mmol). The reaction
was concentrated and the crude product was purified by flash chromatography on silica
gel (gradient elution: 1-3-5% methanol: dichloromethane) to afford 50mg (58%) of a
brown solid: Example 46: mp 285-290°C; MS (ES+calculated: 412.29; found: 412.48,
414.04 M+H). HPLC (85%) purity, retention time 12.686 minutes - Method B); 1H
Example 47
3-(2-cyclohexyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-fluoro-1,3-dihydro-indol-2-one
In a similar fashion as for the preparation of 45 above, 5-fluorooxindole (Combiblocks,
32mg, 0.21 mmol) was reacted with Compound 23 (50mg, 0.21 mmol). The reaction
was concentrated and the crude product was purified by flash chromatography on silica
gel (gradient elution: 1-3-5% methanol: dichloromethane) to afford 39mg (53%) of a
yellow solid. Example 47: mp 294-298°C; HPLC (88%) purity, retention time 12.604
minutes - Method B); 1H NMR (400 MHz, trifluoroacetic acid-d) d 8.72 (s), 8.54 (s),
7.13 (d, J=8 Hz), 6.88 (m), 6.78 (m), 3.72 (s), 2.15-0.85 (m, 10H).
Example 48
5-Chloro-3-(2-cyclohexyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
In a similar fashion as for the preparation of 45 above, 5-chlorooxindole (Combiblocks,
35mg, 0.21 mmol) was reacted with Compound 23 (50mg, 0.21 mmol). The reaction
was concentrated and the crude product was purified by flash chromatography on silica
gel (gradient elution: 1-3-5% methanol: dichloromethane) to afford 22mg (28%) of an
orange-yellow solid. Example 48: mp 294-298°C; MS (ES+calculated: 367.84; found:
368.36 M+H). HPLC (78%) purity, retention time 13.406 minutes - Method B); 1H
NMR (400 MHz, trifluoroacetic acid-d) d 8.97 (s), 8.78 (s), 7.64 (s), 7.18 (d, J=8 Hz),
7.10 (d, J=8 Hz), 4.43 (m), 2.301.11 (m, 10H).
Example 49
3-(2-Cyclohexyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-nitro-1,3-dihydroindol-2-one
In a similar fashion as for the preparation of 45 above, 5-nitrooxindole (Combiblocks,
30mg, 0.17 mmol) was reacted with Compound 23 (40mg, 0.17 mmol). The reaction
was concentrated and the crude product was purified by flash chromatography on silica
gel (gradient elution: 1-3-5% methanol: dichloromethane) to afford 18mg (28%) of a
yellow-brown solid. Example 49: mp 301-304°C; MS (ES+calculated: 378.39; found:
379.25 M+H). HPLC (100%) purity, retention time 12.798 minutes - Method B); 1H
NMR (400 MHz, trifluoroacetic ncid-d) d 9.21 (s, 1H), 9.00 (s, 1H), 8.73 (s, 1H), 8.42
(s, 1H), 7.52 (s, 1H), 5.28 (s, 1H), 4.62 (m, 1H), 2.56-1.11( m, 10H).
Example 50
3-(2-cyclohexyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-6-fluoro-1,3-dihydro-indol-2-one
In a similar fashion as for the preparation of 45 above, 6-fluorooxindole (Combiblocks,
32mg, 0.21 mmol) was reacted with Compound 23 (50mg, 0.21 mmol). The reaction
was concentrated and the crude product was purified by flash chromatography on silica
gel (gradient elution: 1-3-5% -methanol: dichloromethane) to afford 22mg (30%) of an
orange solid. Example 50: mp 297-300°C; MS (ES+calculated: 351.39; found: 352.18
M+H). HPLC (89%) purity, retention time 12.457 minutes - Method B); 1H NMR (400
MHz, trifluoroacetic acid-d) d 9.00 (s, 1H), 8.79 (s, 1H), 7.70 (s, 1H), 6.97 (m, 2H),
4.48 (m, 1H), 2.47-1.24 (m, 10H).
Example 51
6-Chloro-3-(2-cyclohexyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
In a similar fashion as for the preparation of 45 above, 6-chlorooxindole (Combiblocks,
35mg, 0.21 mmol) was reacted with Compound 23 (50mg, 0.21 mmol). The reaction
was concentrated and the crude product was purified by flash chromatography on silica
gel (gradient elution: 1-3-5% methanol: dichloromethane) to afford 19mg (25%) of an
orange solid. Example 51: mp 299-303oC; MS (ES+calculated: 367.84; found: 368.38
M+H). HPLC (97%) purity, retention time 13.852 minutes - Method B); 1H NMR (400
MHz, trifluoroacetic acid-d) d 9 00 (s, 1H), 8.77 (s, 1H), 7.63 (m, 1H), 7.28 (m, 2H),
4.50 (m, 1H), 2.50-1.30 (m, 10H)
Example 52
3-(2-Cyclohexyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-methoxy-1,3-dihydro-indol-2-
one
In a similar fashion as for the preparation of 45 above, 5-methoxyoxindole (36mg, 0.22
mmol) was reacted with Compound 23 (50mg, 0.21 mmol). The reaction was
concentrated and the crude product was purified by flash chromatography on silica gel
(gradient elution: 1-2-3% methanol: dichloromethane) to afford 17mg (22%) of a
yellow solid. Example 52: MS (ES+calculated: 363.42; found: 364.18 M+H). HPLC
(62%) purity, retention time 12.048 minutes - Method B); 1H NMR (400 MHz, DMSO-
d6) d 10.54 (s, 1H), 8.88 (s, 1H), 8.29 (s, 1H), 7.26 (s, 2H), 6.83 (d, J=8 Hz, 1H), 6.69
(d, J=8 Hz, 1H), 4.56 (m, 1H), 2.::0-1.15 (m, 10H).
Example 53
3-(2-Cyclohexyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-trifluoromethyl-1,3-dihydro-
indol-2-one
In a similar fashion as for the preparation of 45 above, 5-trifluoromethyloxindole
(Combiblocks, 44mg, 0.22 mmol) was reacted with Compound 23 (50mg, 0.21 mmol).
The reaction was concentrated and the crude product was purified by flash
chromatography on silica gel (gradient elution: 1-5% methanol: dichloromethane) to
afford 58mg (69%) of a yellow solid. Example 53: mp >300°C; MS (ES+calculated:
401.39; found: 402.20 M+H). HPLC (92%) purity, retention time 14.161 minutes -
Method B); 1H NMR (400 MHz, trifluoroacetic acid-d) d 9.10 (s, 1H), 8.84 (s, 1H),
8.03 (s, 1H), 7.68 (s, 1H), 7.40 (s, 1H), 4.50 (m, 1H), 2.60-1.23 (m, 10H).
Example 54
5-Bromo-3-(1-cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
To 5-bromooxindole (Combi-Blocks, 136mg, 0.64mmol) and N,N,N',N'-
tetramethylethylenediamine (Acros, 0.193mL, 1.28mmol) in 10 mL anhydrous THF
under argon at -78°C was added lithium diisopropylamine (Acros, 0.64mL of a 2.0M
solution in THF/heptane, 1.28nunol). The solution was stirred for fifteen minutes at
which point a solution of Compound 25 (150mg, 0.676mmol) in 10mL THF was added
dropwise at such a rate as to maintain the temperature below -50°C. The reaction was
stirred for ten minutes, external cooling was removed, and the reaction was permitted to
warm to room temperature. An oil bath was applied and the mixture was refluxed for 6
hours. The reaction was concentrated. The residue was dissolved in dichloromethane
and applied to flash chromatography on silica gel (gradient elution: 1-3% methanol:
dichloromethane). Fractions determined to contain 54 by LC/MS were concentrated and
dried in vacuo to afford 80mg (30%) of an orange solid. Example 54: mp 287-290°C,
270 °C (soft); MS (ES+calculate I: 398.27; found: 398.39,399.85 M+H). HPLC (88%)
Example 55
3-(1-Cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
Using the procedure outlined for 54, 5-cyanooxindole (Combi-Blocks, 116mg,
0.736mmol) and Compound 25 (I72mg, 0.775mmol) were refluxed overnight affording
30mg (11%) of yellow crystals. Example 55: mp >300°C; MS (ES+calculated: 344.38;
found: 345.21 M+H). HPLC (95%) purity, retention time 4.946 minutes - Method C);
Example 56
3-(1-Cyclopentyl-1H-pyrazolo[2,4-d]pyrinndine-4-yl)-5-methyl-1,3-dihydro-indol-2-
one
Using the procedure outlined for 54, 5-trifluoromethyloxindole (Combi-Blocks, 68mg,
0.338 mmol) and Compound 25 (75mg, 0.338mmol) were refiuxed overnight affording
25mg (19%) of yellow solid. Example 56: mp >300°C; MS (ES+calculated: 387.37;
found: 388.19 M+H). HPLC (96%) purity, retention time 6.279 minutes - Method C);
Example 57
3 -(1-Cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-nitro-1,3-dihydro-indol-2-one
Using the procedure outlined for 54, 5-nitrooxindole (Combi-Blocks, 60.2mg,
0.338mmol) and Compound 25 (75mg, 0.338mmol) were refluxed overnight affording
25mg (20%) of orange solid Example 57: mp >300 °C (dec); MS (ES+calculated:
364.37; found: 365.19 M+H). HPLC (85%) purity, retention time 5.504 minutes -
Example 58
5-Chloro-3-(1-cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Using the procedure outlined :br 54, 5-chlorooxindole (Aldrich, 56.6mg, 0.338mmol)
and Compound 25 (75mg, 0.328mmol) were refluxed overnight affording 17mg (14%)
of yellow solid. Example 58: mp 304-306 °C; MS (ES+calculated: 353.81; found:
Example 59
6-Chloro-3-(1-cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3 -dihydro-indol-2-one
Using the procedure outlined for 54, 6-chlorooxindole (Combi-Blocks, 56.6mg,
0.338mmol) and Compound 25 (75mg, 0.338mmol) were refluxed overnight affording
Example 60
3-(1-cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-5,7-dinitro-1,3-dihydro-indol-2-
one
Using the procedure outlined for 54, 5,7-dinitrooxindole (Combi-Blocks, 100mg,
0.45mmol) and Compound 25 (100mg, 0.45mmol) were refluxed overnight affording
7mg (4%) of yellow solid. Example 60: mp 242-246°C; MS (ES+calculated: 409.36;
found: 410.24 M+H). HPLC (95%) purity, retention time 6.416 minutes - Method C);
1H NMR (400 MHz, DMSO-d6) d 13.15 (br s, 1H), 9.02 (s, 1H), 8.81 (s, 1H), 8.65 (s,
1H), 8.40 (s, 1H), 6.95 (s, 1H), 5.26 (m, 1H), 2.18 (m, 2H), 2.05 (m, 2H), 1.92 (m, 2H),
1.75 (m,2H).
Example 61
3-(1-cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-5,7-difluoro-1,3-dihydro-indol-2-
one
Using the procedure outlined for 54, 5,7-difluorooxindole (Oakwood, 76mg, 0.45mmol)
and Compound 25 (100mg, 045mmol) were refluxed overnight affording 20mg (13%)
of bright yellow solid. Example 61: mp >300°C; MS (ES+calculated: 355.35; found:
Example 62
3-(1Cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Using the procedure outlined for 54, oxindole (Combi-Blocks, 59.9mg, 0.45mmol) and
Compound 25 (100mg, 0.45mmol) were refluxed overnight affording 32mg (22%) of
yellow solid. Example 62: mp 228-231°C; MS (ES+calculated: 319.37; found: 320.16
Example 63
3-(1-Cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)2-oxo-2,3-dihydro-1H-indole-6-
carbonitrile
Using the procedure outlined for 54, 6-cyanooxindole (Combi-Blocks, 80mg,
0.505mmol) and Compound 25 filling, 0.5mmol) were refluxed overnight affording
75mg (44%) of orange solid. Example 63: mp 335-338°C; MS (ES+calculated: 344.38;
found: 345.19 M+H). HPLC (94%) purity, retention time 12.942 minutes - Method B):
Example 64
3-(1-Cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)2-oxo-2,3-dihydro-1H-indole-7-
carbonitrile
Using the procedure outlined for 54, 7-cyanooxindole (Combi-Blocks, 80mg,
0.505mmol) and Compound 25 (11mg, 0.5mmol) were refluxed overnight affording
47mg (27%) of orange solid. Example 64: mp 327-332°C; MS (ES+calculated: 344.38;
found: 345.18 M+H). HPLC (95%) purity, retention time 13.114 minutes - Method B);
Example 65
3-(1-Cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-fluoro-1,3-dihydro-indol-2-one
Using the procedure outlined for 54, 5-fluorooxindole (Combi-Blocks, 76mg,
0.503mmol) and Compound 25 (11 mg, 0.5mmol) were refluxed overnight affording
25mg (15%) of orange-yellow solid. Example 65: mp 254-257°C; MS (ES+calculated:
337.36; found: 338.21 M+H). HPLC (80%) purity, retention time 13.366 minutes -
Example 66
3-(l -Cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-6-fluoro-1,3-dihydro-indol-2-one
Using the procedure outlined for 54, 6-fluorooxindole (Combi-Blocks, 76mg,
0.503mmol) and Compound 2:5 (111mg, 0.5mmol) were refluxed overnight affording
42mg (25%) of orange-yellow solid. Example 66: mp 230-233°C; MS (ES+calculated:
337.36; found: 338.24 M+H). HPLC (77%) purity, retention time 13.591 minutes -
Example 67
3-(1-Cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-4,5-difluoro-1,3-dihydro-indol-2-
one
Using the procedure outlined for 54, 4,5-difluorooxindole (Combi-Blocks, 85mg,
0.503mmol) and Compound 25 (111mg, 0.5mmol) were refluxed overnight affording
27mg (15%) of yellow solid. Example 67: mp >300°C; MS (ES+calculated: 355.35;
found: 356.20 M+H). HPLC (90%) purity, retention time 13.323 minutes - Method B);
!H *1H NMR (400 MHz, DMSO d6) d 10.87 (br s, 1H), 8.49 (s, 1H), 8.16 (d, J=8 Hz,
1H), 7.03 (m, 1H), 6.7 (m, 1H), 5.22 (m, 1H), 2.09 (m, 2H), 2.00 (m, 2H), 1.89 (m, 2H),
1.70 (m,2H).
Example 68
3-(1-Cyclohexyl-1H-pyrazolo[3,4-de]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
To 5-cyanooxindole (Comb-Blocks, 67mg, 0.424mmol) and N,N,N',N'-
tetramethylethylenediamine (Acros, 0.128mL, 0.848mmol) in 5 mL anhydrous THF
under argon at -78°C was added lithium diisopropylamine (Acros, 0.424mL of a 2.0M
solution in THF/heptane, 0.848nunol). The solution was stirred for fifteen minutes at
which point a solution of Compound 26 (100mg, 0.424mmol) was added as a solid. The
reaction was stirred for ten minutes, external cooling was removed, and the reaction was
permitted to warm to room temperature. An oil bath was applied and the mixture was
refluxed overnight. The reaction was concentrated. The residue was dissolved in
dichloromethane and applied to flash chromatography on silica gel (gradient elution: 1-
3% methanol: dichloromethane). Fractions determined to contain 68 by LC/MS were
concentrated and the resulting solid was triturated in methanol and filtered. The solid
was dried in vacuo to afford 60mg (39%) of a yellow solid. Example 68: mp >300°C;
MS (ES+calculated: 358.41; found: 359.19 M+H). HPLC (96%) purity, retention time
Example 69
3-(1-Cyclohexyl-1H-pyrazolo[3,4-d]pyrimidin-4yl)-2-oxo-2,3-dihydro-1H-indole-6-
carbonitrile
Using the procedure outlined for 68, 6-cyanooxindole (Combi-Blocks, 67mg,
0.424mmol) and Compound 26 (100mg, ().424mmol) were refluxed overnight affording
30mg (20%) of dark yellow solid. Example 69: mp >300°C; MS (ES+calculated:
358.41: found: 359.22 M+HY HPLC (98%) purity retention time 5.526 minutes -
Example 70
3-(1-Cyclohexyl-1H-pyrazolo[3,4-d]pyrimidin-4yl)-2-oxo-2,3-dihydro-1H-indole-7-
carbonitrile
Using the procedure outlined for 68, 7-cyanooxindole (Combi-Blocks, 67mg,
0.424mmol) and Compound 26 (100mg, 0.424mmol) were refluxed overnight affording
27mg (18%) of yellow solid. Example 70: mp >300°C; MS (ES+calculated: 358.41;
found: 359.28 M+H). HPLC (96%) purity, retention time 5.814 minutes - Method C);
1H NMR (400 MHz, DMSO-d6) d 14.58 (br s, 1H), 11.65 (s, 1H), 8.67 (s, 1H), 8.55 (s,
1H), 8.08 (d, J=8 Hz, 1Hz), 7.39 (d, J=8 Hz, 1H), 7.15 (t, J=8 Hz), 4.71 (m, 1H), 1.91
(m, 6H), 1.71 (m, 1H), 1.47 (m, 2H), 1.29 (m, 1H).
Example 71
3-(1-Cyclohexyl-1H-pyrazole[3,4-d]pyrimidin-4-yl)-5-trifluoromethyl-1,3-dihydro-
indol-2-one
Using the procedure outlined for 68, 5-trifluoromethyloxindole (Combi-Blocks, 85mg,
0.424mmol) and Compound 26 (100mg, 0.424mmol) were refluxed overnight affording
45mg (26%) of bright yellow solid. Example 71: mp >300°C; MS (ES+calculated:
401.39; found: 402.25 M+H). HPLC (93%) purity, retention time 6.578 minutes -
Example 72
3-(1-Cyclohexyl-1H-pyrazolo[34-d]pyrimidin-4-yl)-5-fluoro-1,3-dihydro-indol-2-one
Using the procedure outlined for 68, 5-fluorooxindole (Combi-Blocks, 64mg,
0.424mmol) and Compound 26 (100mg, 0.424mmol) were refluxed overnight affording
50mg (34%) of yellow solid. Example 72: mp 282-286°C; MS (ES+calculated: 351.39;
found: 352.21 M+H). HPLC (98%) purity, retention time 5.780 minutes.- Method C):
Example 73
3-(1-Cyclohexyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-6-trifluoromethyl-1,3-dihydro-
indol-2-one
Using the procedure outlined for 68., 6-fluorooxindole (Combi-Blocks, 64mg,
0.424mmol) and Compound 26 (100mg, 0.424mmol) were refluxed overnight affording
15mg (10%) of bright yellow solid. Example 73: mp 277-280°C; MS (ES+calculated:
351.39; found: 352.18 M+H). HPLC (94%) purity, retention time 5.820 minutes -
Example 74
5-Chloro-3-(1-Cyclohexyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Using the procedure outlined for 68, 5-chlorooxindole (Aldrich, 70.4mg, 0.424mmol)
and Compound 26 (100mg, 0.424mmol) were refluxed overnight affording 21mg (13%)
of yellow solid. Example 74: mp 300-305°C; MS (ES+calculated: 367.84; found:
368.33 M+H). HPLC (97%) purity, retention time 15.299 minutes - Method B); 1H
NMR (400 MHz, DMSO-d6) d 10.36 (br s, 1H), 8.49 (s, 1H), 8.48 (s, 1H), 7.66 (s, 1H),
7.07 (d, J=8 Hz, 1H), 6.93 (d, J=8 Hz, 1H), 4.70 (m, 1H), 1.93-1.28 (m, 10H).
Example 75
5-Bromo-3-(1-Cyclohexyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Using the procedure outlined for 68, 5-bromooxindole (Combi-Blocks, 89mg,
0.424mmol) and Compound 26 (100mg, 0.424mmol) were refluxed overnight affording
50mg (29%) of yellow solid. Example 75: mp 305-308°C; MS (ES+calculated: 412.29;
found: 412.51, 413.84 M+H). HPLC (96%) purity, retention time 15.590 minutes -
Method B); 1H NMR (400 MHz, DMSO-d6) d 10.87 (br s, 1H), 8.50 (s, 1H), 8.45 (s,
1H), 7.78 (s, 1H), 7.20 (d, J=8 Hz, 1H), 6.90 (d, J=8 Hz, 1H), 4.70 (m, 1H), 2.08-1.25
(m, 10H).
Example 76
3-(2-Allyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
In a similar fashion as for the preparation of 45 above, 5-cyanooxindole (Combiblocks,
60mg, 0.38 mmol) was reacted with Compound 24 (77mg, 0.40 mmol). The reaction
was concentrated and the crude product was triturated successively with methanol and
water. A yellow-orange solid was obtained which was dried in vacuo. Yield: 67mg
(56%). Example 76: mp >300°C; MS (ES+calculated: 316.32: found: 317.09 M+H).
Compound 77
2-Cyclopentyl-2,7-dihydro-pyrazolo[3,4-d]pyrimidine-4,6-dione
Compound 12 (1g, 5.15 mmol) was mixed with urea (Fisher, 3g, 50 mmol) and fused at
200°C for ninety minutes. The solution was allowed to cool briefly and 10 mL water
was added. The solution was boiled for one hour. The white solid present was removed
by filtration and was determined to contain additional urea. The solid was recombined
with 20 mL water and resubjected to boiling for an additional hour. On cooling and
filtration, a white solid was obtained which was dried in vacuo. Yield: 837mg, (74%).
Compound 77: mp >300°C; MS (ES+calculated: 220.23; found: 221.17 M+H). HPLC
(92%) purity, retention time 5.112 minutes - Method B); 1H NMR (400 MHz, DMSO-
d6) d 11.30 (br s, 1H), 10.63 (br s, 1H), 8.37 (s, 1H), 4.69 (m, 1H), 2.06 (m, 2H),
1.92(m, 2H), 1.77 (m, 2H), 1.63 (m, 2H).
Scheme 2
Compound 79
4-Chloro-1H-pyrazolo[3,4-d]pyrimidine
To a mixture of commercially available: 4-hydroxypyrazolo[3,4-d]pyrimidine (78)
(Acros, 14.5g, 106.5mmol) stirred in POCl3 (375mL) was added N,N-dimethylaniIine
(21mL). The mixture was refluxed for 1.5h. After cooling, excess POCl3 was removed
by rotarty evaporation and pumped on high vacuum before pouring over 500mL ice
while stirring. The mixture was stirred for 10min before extracting with ethyl ether (6 x
250mL). The combined organic layer was washed with ice water (3 x 100mL) and dried
over MgSO4 and filtered. The ethyl ether was stripped and the resulting pale yellow
solid (10g, 61%) was pumped on high vacuum overnight. Compound 79: mp >300°C,
dec. 125°C; MS (ES+ calculated: 154.56; found: 156.21 M+H). HPLC (98% purity,
retention time 6.033 minutes - method D) '1H NMR (400 MHz, DMSO-d6) d 14.55 (bs,
1H), 8.84 (s, 1H), 8.46 (s, 1H).
Compound 80
4-Chloro-2-methyl-2H-pyrazolo[3,4-d]pyrimidine
Compound 81
4-Chloro-l-methyl-2H-pyrazolo[3,4-d]pyrimidine
Cesium carbonate (Acros, 2.12 g, 6.51 mmol) was added to compound 79 (100g, 5.9
mmol) in N,N-dimethylformamide (Acros, 30 mL) at 0°C followed immediately by
methyl iodide (Acros, 1.01g, 7.1 mmol). The mixture was stirred for three hours.
Cesium carbonate was removed by filtration and the filter cake was washed with a small
amount of DMF. The filtrate and washings were concentrated and the reaction mixture
was subjected to flash chromatography on silica gel (gradient elution 9:1 to 4:1 to 0:1
dichloromethane:ethyl acetate) to afford two white solids: Compound 80 (220mg, 22%)
elutes second and Compound 81 (663mg, 67%) elutes first. Compound 80: mp 196-
200°C; MS (ES+calculated: 168 59; found: 169.57 M+H). HPLC (100% purity,
retention time 4.627 minutes - Method B); 1H NMR (300 MHz, DMSO-d6): 8.91 (s,
1H), 8.90 (s, 1H), 4.25 (s, 3H). Compound 81: mp 97-99°C ; MS (ES+calculated:
168.59; found: 169.37 M+H). EPLC (100% purity, retention time 6.582 minutes -
Method B); 1H NMR (400 MHz, DMSO-d6) d 8.98 (s, 1H), 8.48 (s, 1H), 4.09 (s, 3H).
Example 82
5-Bromo-3-(2-methyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
5-Bromooxindole (Combiblocks, 63 mg, 0.30 mmol) in 10 mL anhydrous
tetrahydrofuran (Acros) at -78°C was treated dropwise with lithium diisopropylamide
(Acros, 0.3 mL of a 2.0M solution in THF/heptane, 0.60 mmol) and the reaction mixture
was stirred for fifteen minutes. Compound 80 (50 mg, 0.30 mmol) in 5 mL anhydrous
tetrahydrofuran was added dropwise and the reaction was permitted to warm to room
temperature over four hours. The reaction was concentrated and purified by flash
chromatography on silica gel (gradient elution, 1-3-5-10-20%
methanol.dichloromethane to 1:20:79 ammonium hydroxide:methanol:dichloromethane)
to afford 82 as a yellow solid (81 mg, 78%). Example 82: mp 360-5°C, MS
(ES+calculated: 344.17; found: 345.85 M+H). HPLC (96% purity, retention time 9.548
minutes - Method B); 1H NMR (400 MHz, DMSO-d6) d 10.90 (s), 9.48 (s), 8.89 (s),
8.34 (s), 7.74 (s), 7.22 (d, J=8 Hz), 6.89 (d, J=8 Hz), 4.17 (s, 3H).
Example 83
5-Chloro-3-(2-methyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Preparation of Example 83: In an identical fashion as for the synthesis of Example 82,
Example 83 was prepared as a yellow solid from compound 80 in quantitative yield.
Example 83: mp 341-4°C, MS (ES+calculated: 299.72; found: 300.16 M+H). HPLC
(100% purity, retention time 9.323 minutes - Method B); 1H NMR (400 MHz, DMSO-
d6) d 10.89 (s), 9.48 (s), 8.93 (s), 8.34 (s), 7.64 (s), 7.09 (d, J=8 Hz), 6.93 (d, J=8 Hz),
4.17 (s,3H).
Example 84
3-(2-Methyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
In an identical fashion as for the synthesis of Example 82, Example 84 was prepared as
a yellow solid from compound 80 in 50% yield. Example 84: mp >350°C, MS
(ES+calculated: 290.29; found: 291.07 M+H). HPLC (100% purity, retention time
7.467 minutes - Method B); 1H NMR (400 MHz, DMSO-d6) d 11.27 (s), 9.49 (s), 9.11
(s), 8.39 (s), 7.99 (s), 7.50 (d, J=8 Hz), 7.08 (d, J=8 Hz), 4.19 (s, 3H).
Example 85
5-Bromo-3-(1-methyl-1H-pyrazolo[3,4- d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
In an identical fashion as for the synthesis of 82, Example 85 was prepared as a yellow
solid from compound 81 in 20% yield. Example 85: mp >350°C, MS (ES+calculated:
344.17; found: 345.86 M+H). HPLC (95% purity, retention time 11.124 minutes -
Method B); 1H NMR (400 MHz, DMSO-d6) d 10.91 (s), 8.51 (s), 8.43 (s), 7.77(s), 7.20
(d, J=8 Hz), 6.90 (d, J=8 Hz), 3.99 (s, 3H), 3.93 (s).
Example 86
5-Chloro-3-(1-methyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
In an identical fashion as for the synthesis of 82, Example 86 was prepared as a yellow
solid from compound 81 in 49% yield. Example 86: mp >320°C, MS (ES+calculated:
299.72; found: 300.09 M+H). HPLC (99% purity, retention time 10.772 minutes -
Method B); 1H NMR (400 MHz, DMSO-d6) d 10.90 (s), 8.52 (s), 8.47 (s), 7.65(s), 7.08
(d, J=8 Hz), 6.94 (d, J=8 Hz), 3.99 (s, 3H), 3.93 (s).
Compound 87 and 88
2-(propyl-3-azido)- 4-Chloro-2H-pyrazolo[3,4-d]pyrimidine
1-(propyl-3-azido)-4-Chloro-1H-pyrazolo[3,4-d]pyrimidine
In an identical fashion as for the alkylation of 79 with iodomethane, Compound 87 and
Compound 88 were prepared as yellow solids in 10% and 40% yields, repectively.
Compound 87: MS (ES+calculated: 237.65; found: 238.29 M+H). HPLC (100% purity,
Example 89
3-[2-(3-azido-propyl)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-bromo-1,3-dihydro-indol-2-
one
In an identical fashion as for the preparation of Example 82, Compound 87 was
alkylated to produce Example 89 m 91% yield. Compound 89: mp 252-6°C, MS
(ES+caIculated: 413.24; found: 414.87 M+H). HPLC (97% purity, retention time
11.337 minutes - Method B); 1H NMR (400 MHz, DMSO-d6) d 0.90 (s), 9.53 (s), 8.93
(s), 8.34 (s), 7.75 (s), 7.22 (d, J=8 Hz, 1H), 6.89 (d, J=8 Hz, 1H), 4.53-4.38 (m, 2H),
3.44-3.38 (m, 2H), 2.17-2.11 (m, 2H).
Example 90
3-[2-(3-methylamino-propyl)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-bromo-1,3-dihydro-
indol-2-one
Example 89 (20mg, 48.4µmol) was suspended in 5 mL anhydrous 1,2-dichloroethane
(Acros) and treated with bromodimethylborane (Acros, 5µL, 50 (µmol). The mixture
was stirred overnight. Additional bromodimethylborane (in 50 µmol aliquots) was
added on three occasions over three days with a drop of N-methylpyrrolidinone (Acros,
anhydrous) being added on each occasion to promote solubility. The reaction was
concentrated and ethanol was adtled (5 mL). The reaction was stirred ten minutes and
reconcentrated. The solid was taken up into a small amount of methanol (approximately
1-2 mL) and dichloromethane was added to turbidity. The solution was allowed to sit
for several hours and a yellow solid was isolated by filtration. The solid was washed
with a small amount of anhydrous dichloromethane and was dried in vacuo. Yield: 12
mg (51%) of the hydrobromide salt. Example 90: mp 240-3°C, MS (ES+calculated:
401.27; found: 402.90 M+H). HPLC (87% purity, retention time 8.813 minutes -
Method A); 1H NMR (400 MHz, DMSO-d6) d 10.92 (s), 9.537 (s), 8.92 (s), 8.36 (s),
7.75 (s), 7.23 (m), 7.14 (m), 6.90 (n), 6.77 (m), 4.55-4.42 (m), 2.92 (m), 2.51 (m), 2.32-
2.18 (m).
Compound 91 and 92
4-Chloro-1-propyl-1H-pyrazolo[3,4-d]pyrimidine
4-Chloro-2-propyl-1H-pyrazolo[3,4-d]pyrimidine
Cs2CO3 was added to a solution of 79 (2.0g, 13mmol) in 60mL anhydrous DMF at 0°C.
1-Iodopropane (Acros, 1.52mL, 15.6mmol) was added to the suspension and allowed to
stir for 3.5h at 0°C. The reaction mixture was filtered and the solid was washed with
dichloromethane. The filtrate was concentrated to dryness and the residue was
dissolved in 9:1 hex/EtOAc and applied to flash chromatography on silica gel (gradient
elution: 9:1, 4:1, 0:1 hexane: ethylacetate). Fractions determined to contain 91 and 92
by LC/MS were concentrated to afford 830mg (33%) of clear colorless oil 91 and
296mg (12%) yellow solid 92. Compound 91: MS (ES+ calculated: 196.64; found:
197.09 M+H). HPLC (100% purity, retention time 10.314 minutes - method D) 1H
NMR (300MHz, DMSO-d6): 8.87 (s, 1H), 8.48 (s, 1H), 4.45 (t, J=7Hz, 2H), 1.91 (m,
2H), 0.85 (t, J=7, 3H). Compound 92: mp 93-95°C; MS (ES+ calculated: 196.64; found:
197.09 M+H). HPLC (90% purity, retention time 7.931 minutes - method D) 1H NMR
(400 MHz, DMSO-d6) d 8.67 (s, 1H), 8.53 (s, 1H), 4.37 (t, J=7,2H), 1.92 (m, 2H), 0.83
(t,J=7,3H).
Example 93
3-(1-propyl-1H-pyrazolo[3,4-d]pyrimidin-4yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
5-cyanooxindole (Combiblocks, 161mg, 1.02 mmol) in 5 mL anhydrous tetrahydrofuran
was stirred under argon. The solution was cooled to -78ºC and lithium
diisopropylamide (Acros, 1.02 mL of a 2.0M solution in THF/hexane, 2.04 mmol) was
added dropwise. The reaction wa:: stirred for fifteen minutes at which point Compound
91 (200mg, 1.02 mmol) was added. The reaction was stirred an additional fifteen
minutes and was warmed to room temperature over 0.5 hour. The mixture was then
stirred overnight at room temperature. The reaction was concentrated and subjected to
flash chromatography on silica gel (gradient elution: 1-3-5% methanol:
dichloromethane) to afford 30mg (9%) of a yellow solid after trituration in MeOH.
Example 93: mp >300°C; MS (ES+calculated: 318.34; found: 319.18 M+H). HPLC
(99%) purity, retention time 4.068 mnutes - Method C); 1H NMR (400 MHz, DMSO-
d6) d 11.25 (s, 1H), 8.71 (s, 1H), 8.55 (s, 1H), 8.02 (s, 1H), 7.48 (d, J=8, 1H), 7.07 (d,
J=8,1H), 4.35 (t, 2H), 1.88 (m, 2H), 0.87 (t, J=7, 3H).
Example 94
5-Bromo-3-(1-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 93, 5-bromooxindole (Combiblocks, 108mg,
0.51mmol) and Compound 91 (100mg, 0.51mmol) were stirred overnight to afford
86mg (45%) of a yellow solid Example 94: mp 295-300°C; MS (ES+calculated:
372.23; found: 372.47 M+H). HPLC (99%) purity, retention time 5.243 minutes -
Method C); 1H NMR (400 MHz, DMSO-d6) d 14.58 (s), 10.90 (s, 1H), 8.51 (s, 1H),
8.45 (s, 1H), 7.78 (s, 1H), 7.20 (d, 1H), 6.90 (d, J=8, 1H), 4.33 (t, 2H), 1.88 (m, 2H),
0.87 (t, J=7, 3H).
Example 95
5-Chloro-3-(1-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 93, 5-chlorooxindole (Combiblocks, 85.5mg,
0.51mmol) and Compound 91 (100mg, 0.51mmol) were stirred overnight to afford
62mg (37%) of a yellow solid. Example 95: mp 293-297°C; MS (ES+calculated:
327.78; found: 328.28 M+H). HPLC (99%) purity, retention time 5.056 minutes -
Method C); 1H NMR (400 MHz, DMSO-d6) d 14.61 (s), 10.89 (s, 1H), 8.49 (d, J=7,
2H), 7.65 (s, 1H), 7.08 (d, J=8, 1H), 6.93 (d, J=8), 4.33 (t, J=7, 2H), 1.88 (m, 2H), 0.87
(t,J=7,3H).
Example 96
3-(2-propyl-1H-pyrazolo[3,4-d ]pyrimidin-4yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
5-cyanooxindole (Combiblocks, 40.4mg, 0.255 mmol) in 1.5 mL anhydrous
tetrahydrofuran was stirred under argon. The solution was cooled to -78°C and lithium
diisopropylamide (Acros, 0.255 mL of a 2.0M solution in THF/hexane, 0.51 mmol) was
added dropwise. The reaction was stirred for fifteen minutes at which point Compound
92 (50mg, 0.255 mmol) was added. The reaction was stirred an additional fifteen
minutes and was warmed to room temperature over 0.5 hour. The mixture was then
stirred overnight at room temperature. The reaction was concentrated and subjected to
flash chromatography on silica gel (gradient elution: 1-3-5% methanol:
dichloromethane) to afford 18 mg (22%) of a yellow solid after trituration in MeOH.
Example 96: mp >300°C; MS (ES+calculated: 318.34; found: 319.14 M+H). HPLC
(98%) purity, retention time 3.532 minutes - Method C); 1H NMR (400 MHz, DMSO-
d6) d 11.25 (s), 9.52 (s), 9.13 (s), 8.40 (s), 7.98 (s), 7.49 (d), 7.09 (d), 4.40 (m), 4.36 (bs),
4.10 (bs), 1.90 (m), 0.88 (m).
Example 97
5-Bromo-3-(2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Using the procedure outlined for 96, 5-bromooxindole (Combiblocks, 54mg,
0.255mmol) and Compound 92 (50mg, 0.255mmol) were stirred overnight to afford
35mg (37%) of an orange solid. Example 97: mp >300°C; MS (ES+calculated: 372.23;
found: 373.79 M+H). HPLC (98%) purity, retention time 4.271 minutes - Method C);
1H NMR (400 MHz, DMSO-d6) d 14.09 (s), 10.89 (s), 9.50 (s). 8.90 (s), 8.34 (s), 7.73
(s), 7.23 (d), 6.90 (d), 4.41 (t, J=7), 1.93 (m), 0.86 (m).
Example 98
5-Chloro-3-(2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Using the procedure outlined for 96, 5-chlorooxindole (Combiblocks, 42.7mg,
0.255mmol) and Compound 92 (50mg, 0.255mmol) were stirred overnight to afford
55mg (66%) of a yellow solid. Example 98: mp 302-306°C; MS (ES+calculated:
327.78; found: 328.27 M+H). HPLC (98%) purity, retention time 4.145 minutes -
Method C); 1H NMR (400 MHz DMSO-d6) d 10.88 (s), 9.52 (s), 8.95 (s), 8.35 (s), 7.62
(s), 7.10 (d), 6.95 (d), 4.42 (t, J=7), 1.93 (m), 0.90 (m).
Example 99
3-(1-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-trifluoromethyl-1,3-dihydro-indol-2-
one
Using the procedure outlined for 96, 5-trifluoromethyloxindole (Combiblocks, 36mg,
0.179 mmol) and Compound 92 (35mg, 0.179mmol) were stirred overnight to afford
35mg (54%) of a white solid. Example 99: mp 293-295°C; MS (ES+calculated: 361.33;
found: 362.17 M+H). HPLC (98%) purity, retention time 4.145 minutes - Method C);
1H NMR (400 MHz, DMSO-d6) d 11.12 (s), 9.53 (s), 8.85 (s), 8.35 (d), 8.06 (t), 7.87 (s),
7.40 (d), 7.10 (d), 4.23 (m), 1.87 (m), 0.87 (m).
Example 100
3-(1-propyl-2H-pyrazolo[3,4-d]pyrimidm-4-yl)-5-fluoro-1,3-dihydro-indol-2-one
Using the procedure outlined for 96, 5-fluorooxindole (Combiblocks, 27mg,
0.179mmol) and compound 92 (35mg, 0.179mmol) were stirred overnight to afford
30mg (54%) of a yellow solid. Example 100: mp 288-292°C; MS (ES+calcuIated:
311.32; found: 312.13 M+H). HPLC (93%) purity, retention time 3.762 minutes -
Method C); 1H NMR (400 MHz, DMSO-d6) d 14.32 (s), 10.79 (s), 9.03 (s), 8.34 (s),
7.50 (d, J=10), 6.91 (s), 4.42 (s), 1.92 (s), 0.89 (m).
Compounds 101 and 102
3-Amino-1-propyl-1H-pyrazole-4-carbonitrile
5-Amino-1-propyl-1H-pyrazole-4-carbonitrile
3-Amino-4-pyrazolecarbonitrile I (Acros, 3.24g, 30.0 mmol), propylbromide (Acros,
4.43g, 36 mmol) and anhydrou;: potassium carbonate (Fisher, 5.0g, 36 mmol) were
suspended in 20 mL anhydrous DMF and heated at 80°C in a sealed tube under argon
overnight. The reaction was permitted to cool and the DMF was removed on a rotary
evaporator. Water was added (100 mL) and the organics were extracted with
dichloromethane (3 X 100 mL). The combined dichloromethane fractions were washed
with water (50 mL) and brins (50 mL) and were dried (magnesium sulfate).
Concentration of the organics afforded an oil which was subjected to flash
chromatography on silica gel (1-3% methanol/dichloromethane). Two white crystals
were obtained: compound 101 (l.88g, 42%) elutes first and compound 102 (711mg,
16%) elutes second. Compound 101: mp 85-90°C; MS (ES+calculated: 150.18; found:
151.15 M+H). HPLC (99%) purity, retention time 5.8 minutes - Method A); 1H NMR
(400 MHz, DMSO-d6) d 8.07 (s, 1H), 5.52 (s, 2H), 3.82 (t, J=7, 2H), 1.70 (m, 2H), 0.79
(t, J=7, 3H). Compound 102: mp 162-164°C; MS (ES+calculated: 150.18; found:
151.18 M+H). HPLC (95%) purity, retention time 6.4 minutes - Method A); 1H 1H
NMR (400 MHz, DMSO-d6) d 7.51 (s, 1H), 6.53 (s, 2H), 3.82 (t, J=7, 2H), 2.95 (m,
2H), 0.81 (t, J=7,3H).
Compound 103
3-Amino-l-propyl-1H-pyrazole-4-carboxylic acid amide
To concentrated sulfuric acid (Fisher, 1.5 mL) at 0°C was added 101 (931mg,
6.21mmol). The reaction was permitted to warm to room temperature and was stirred
for three hours. At the end of this period all solid had dissolved. This viscous mixture
was then added slowly (violent) o 15 mL concentrated ammonium hydroxide solution
(Fisher). The mixture was stirred for ten minutes and the white solid that formed was
collected by filtration, was washed with water, and was dried in vacuo. The filtrate was
also concentrated, triturated in water and filtered. Both white crystal solids were
compound 103. Yield: 850mg (82%). Compound 103: mp 160-163°C; MS
(ES+calculated: 168.20; found: 169.31 M+H). HPLC (99% purity, retention time 5.358
minutes - Method D); 1H NMR (400 MHz, DMSO-d6) d 7.86 (s, 1H), 7.18 (bs, 1H),
6.70 (bs, 1H), 5.33 (s, 2H), 3.78 (t, J=7, 2H); 1.70 (m, 2H), 0.81 (t, J=7, 3H).
Compound 104
2-Propyl-2H-pyrazolo [3,4-d]pyrimidine-4,6-diol
Compound 103 (510mg, 3.03mmol) and urea (Fisher, 1.5g, 25mmol) were melted at
200°C with stirring. After 1.5h the reaction was allowed to cool to 100°C when 10mL
water was added and the reaction mixture was boiled overnight. The mixture was
cooled and filtered followed by water wash. The white solid was dried in vacuo to
afford 412mg (70%) of compound 104. Compound 104: mp >300°C; MS
(ES+calculated: 194.19; found: 195.19 M+H). HPLC (99% purity, retention time 5.828
minutes - Method D); 1H NMR (400 MHz, DMSO-d6) d 11.34 (s, 1H), 10.66 (s, 1H),
8.33 (s, 1H), 4.05 (t, J=7, 2H), 1.78 (m, 2H), 0.82 (t, J=7, 3H).
Compound 105
4,6-Dichloro-2- propyl-1H-pyrazolo[3,4-d]pyrimidine
A mixture of commercially available 4,6-dihydroxypyrazolo[3,4-d]pyrimidine 104
(Acros, 400mg, 2.06mmol) stirred in POCl3 (10mL) was refluxed overnight. After
cooling, excess POCl3 was removed by rotarty evaporation and pumped on high vacuum
before adding ice chips while stilting. The mixture was stirred for 10min before the
white precipitate was filtered and iried in vacuo. Yield 407mg (85%). Compound 105:
mp 80-84°C; MS (ES+ calculated: 232.10; found: 233.00 M+H). HPLC (99%) purity,
retention time 10.091 minutes - method D) 1H NMR (400 MHz, DMSO-d6) d 9.07 (s,
1H), 4.46 (t, J=7, 2H), 1.95 (m, 2H), 0.85 (t, J=7, 3H).
Compound 107
4,6-Dichloro-5-[l,3]dioxolan-2-yl-pyrimidine
A mixture of 300 ml benzene and 8.2 ml ethylene glycol was heated to reflux and 100
ml solution was distilled off. To the hot solution was added 4,6-dichloro-5-
pyrimidinecarbaldehyde (Bionet, 8.6g, 48.6 mmol) and p-toluenesulfonic acid
monohydrate (Aldrich, 150mg, 0.8 mmol). The mixture was returned to reflux and
water was removed via Dean-Stark trap over 3 h. After cooling, the solvents were
removed under vacuum to yield a dry, yellow solid. The solids were slurried in H2O (30
ml) / saturated NaHCO3 solution (30 ml). White solid 107 was filtered off and dried
under vacuum (8.8g, 82%). Compound 107: mp 108-110°C; MS (ES+calculated:
221.04; found: 221/223 M+H). 1H NMR (400 MHz, DMSO-d6) d 8.95 (s, 1H), 6.26 (s,
1H), 4.23 (m, 2H), 4.05 (m, 2H).
Example 108
3-(1-methyl-1H-pyrazolo[3,4-d]pyrimidin-4yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
A solution of 5-cyanooxindole (Combiblocks, 31.6 mg, 0.2 mmol) in 2 mL anhydrous
tetrahydrofuran under argon was cooled to -78°C and lithium diisopropylamide (Acros,
0.2 mL of a 2.0M solution in THF/hexane, 0.4 mmol) was added dropwise. The
reaction was stirred for fifteen minutes at which point a solution of compound 107
(46.8mg, 0.21 mmol) in 2 ml, anhydrous tetrahydrofuran was added dropwise. The
reaction was stirred an additional fifteen minutes and was wanned to room temperature
for 1 hour. The orange solution was quenched with methylhydrazine (100µl) and stirred
at ambient temperature 16 h. The solvents were removed under vacuum and methanol
(1ml) was added to the solids. After 10 min. of stirring, the tan solids were filtered off
(5.0 mg, 8.6%). Example 108: mp >300°C (dec); MS (ES+calculated: 290.29; found:
291 M+H). 1H NMR (400 MHz, DMSO-d6) d 14.59 (br s, 1H), 11.27 (s, 1H), 8.70(s,
1H), 8.56 (s, 1H), 8.01 (s, 1H), 7.48 (d, J=8.6 Hz, 1H), 7.08 (d, J=8.6 Hz, 1H), 4.01 (s,
3H).
Example 109
2-Oxo-3-(1H-pyrazolo[3,4-d]pyrimidin-4yl)-2,3-dihydro-1H-indole-5-carbonitrile
A solution of 5-cyanooxindole (Combiblocks, 79.0mg, 0.5 mmol) in 5 mL anhydrous
tetrahydrofuran under argon was tooled to -78°C and lithium diisopropylamide (Acros,
0.5 mL of a 2.0M solution in THF/hexane, 1.0 mmol) was added dropwise. The
reaction was stirred for fifteen minutes at which point a solution of compound 107
(113mg, 0.53 mmol) in 5 mL anhydrous tetrahydrofuran was added dropwise. The
reaction was stirred an additional fifteen minutes and was warmed to room temperature
for 1 hour. The orange solution was quenched with hydrazine monohydrate (120µl) and
stirred at ambient temperature 24 h, then at reflux for 16 h. The solvents were removed
under vacuum and methanol (lml) was added to the solids. After 10 min. of stirring, the
tan solids were filtered off. Methanol addition and filtration was repeated 3 times to
give a dark brown solid (21 mg, 15%). Example 109: mp >300°C (dec); MS
(ES+calculated: 276.26; found: 277 M+H). 1H NMR (400 MHz, DMSO-d6) d 14.29
(br s), 11.57 (s), 11.25 (s), 10.01 (br s), 9.45 (s), 9.27 (s), 8.70 (m), 8.50 (s), 8.37 (s),
8.02 (s), 7.82 (s), 7.47 (d, J=9 Hz), 7.06 (d, J=8 Hz), 6.79 (m).
Compound 110
4-Chloro-2-pentyl-2H-pyrazolo[3,4-d]pyrimidine
To a suspension of 79 (3.00 g, 19.41 mmol) and CsCO3 (6.95 g, 21.34 mmol) in 50 mL
of anhydrous DMF was charged pentyliodicle (3.04 mL, 23.29 mmol) drop wise with a
syringe. Allow reaction to stir at room temperature for 36h. After filtering the solids,
the remaining DMF was concentrated in vacuo. The residue was resuspended in water
and dichloromethane, the layers separated and the aqueous layer extracted with 2 x 20
mL portions of dichlormethane. The combined organic was dried over MgS04, filtered
and concentrated in vacuo. The resulting reddish solid was purified via column
chromatography on 150 g SiO2, elated sequentially with the following concentrations of
ethyl acetate in dichloromethane: 10%, 20%,, 50%, 100%. The most polar eluted
compound was collected, stripped to a brown oily solid, (189 mg, 4.3 %) and used in the
next reaction. MS (m/e) 225 (M+l).
Example 111
3-(2-pentyl-1H-pyrazolo[3,4-d]pyrimidin-4yl)-2-oxo-2,3-dihydro-1H-indole-6-
carbonitrile
To the 5-cyanooxindole 110 (71 mg, 0.45 mmol) in 2 mL of anhydrous THF stirring at -
78°C, was added LDA (Acros 2.0m in THF/n-pentane, 560µL, 1.125 mmol) drop wise
with a syringe. The anionic solution was stirred at -78°C for 45 minutes. To the
solution was added the isolated 4-chIoropyrazolo-2-n-pentyl[3,4-d]pyrimidine in 1 mL
anhydrous THF. The solution was stirred at -78°C for 1 hour and then allowed to warm
and stir at room temperature for 2 hours. The reaction was quenched with 2 mL of
saturated NH4Cl, transferred to a separatory funnel and partitioned between
dichloromethane and water. After extracting the water layer with 2 x 20 mL portions of
dichloromethane, the combined organic was dried over MgSO4, filtered and
concentrated in vacuo. The resulting residue was purified via flash chromatography on
60 g SiO2, and eluted with 5% methanol in dichloromethane yielding a yellow solid (44
mg, 28%). Example 111: mp = 290°C dec, 1H NMR (400 MHz, DMSO-d6) d 11.25
(s, 1H), 9.50 (s, 1H), 9.10 (s,lH), 8.35 (d, J= 4 Hz), 7.95 (s, 1H), 7.5 (d, J= 8 Hz, 1H),
7.05 (d, J= 8 Hz, 1H), 4.45 (t, J = 7 Hz, 1H), 4.20 (t, J= 7 Hz, 1H), 1.90 (m, 2H), 1.3
(m, 4H), 0.85 (t, J= 7 Hz, 3H); MS m/e 347 (M+l), HPLC (99% purity, retention time
9.82 minutes, method B).
Compound 112
4,6-Dichloro-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrrnidine
To a 50 mL flask was added compound 77 (500 mg, 2.27 mmol) and POCl3 (8 mL) and
the mixture was heated to reflux for 1.5 h. After cooled to room temperature, the
reaction was concentrated in vacuo. The residue was quenched with ice-water (30 mL)
and basified with NaOH (10N) solution to pH 9. The precipitation was filtered and
washed with water, dried to give 5(4 mg (86%) of the desired compound 112.
Compound 112: 1H NMR (400 MHz, DMSO-d6) d 9.10 (s, 1H), 5.11 (m, 1H), 2.22-
1.71 (m, 8H); MS (m/e) 257 (M + 1).
Example 113
3-(6-Chloro-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-
indole-5-carbonitrile
To a stirring solution of 5-cyanooxindole (435 mg, 2.75 mmol) and THF (25 mL) in 125
mL flask at -78°C was added IDA (3.5 mL, 7.0 mmol). After the reaction was stirred
for 45 min, a solution of compound 112 (707 mg, 2.75 mmol) in THF (5 mL x 2) was
added and continued to stir for 1 h at -78°C. The reaction was allowed to warm to room
temperature and stirred for additional 2 h. It was quenched with water (30 mL) and was
acidified to pH 2 with concentrated HC1. The resulting precipitate was filtered, washed
with water, and dried under house vacuum at 50°C overnight to give 551 mg (53%) of
the desired product Example 113. Example 113: 1H NMR (400 MHz, DMSO-d6) d
10.88 (s, 1H), 9.41 (s, 1H), 8.41 (s, 1H), 7.45 (d, 1H), 6.97 (d, 1H), 5.01 (m, 1H), 2.19-
1.69 (m, 9H); -MS (m/e) 379 (M + 1).
Example 113A
5-Chloro-3-(6-chloro-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-
indol-2-one
Scheme 3 discloses a general procedure for the preparation of compounds of Formula
XI. To a reaction vessel can be added a compound of Formula X, about 10 equivalents
of a compound of formula NHR9R10, and 2-methoxyethanol. The reaction mixture can
be heated to reflux for about 6 to 7 hours. The reaction when complete can be cooled to
room temperature, can be concentrated and the residue can be purified by column
chromatography to give the desirtid product.
Example 114
3-(2-cyclopentyl-6-(2-pyridin-3-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-
oxo-2,3-dihydro-1H-indole-5-carbonitrile
To a Carousel tube was added Example 113 (200 mg, 0.528 mmol), 3-(2-
aminoethyl)pyridine (645 mg, 5.23 mmol), and 2-methoxyethanol (8 mL). The reaction
mixture was heated to reflux for 6.5 h. After cooled to room temperature, the reaction
was concentrated and the residue was purified by Biotage (CH2Cl2/MeOH 10:1) to give
The following Examples 115-144 in Table 1 were prepared according to procedures
disclosed herein, using the appropriate starting materials, including the general
procedure for the preparation of compounds of Formula XI, disclosed herein,
displacement of halogens by amines, alcohols or water, and/or using methods generally
known to one skilled in the art.
Example 115
3-(2-cyclopentyl-6-methylamino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-
1H-indole-5-carbonitrile
Example 116
3-(6-butylamino-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-
1H-indole-5-carbonitrile
Example 117
3-(2-cyclopentyl-6-(4-methyl-piperazin-l-yl)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-
2,3-dihydro-1H-indole-5-carbonitrile
Example 118
3-(2-cyclopentyl-6-morpholin-4-yl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-
dihydro-1H-indole-5-carbonitrile
Example 119
3-(2-cyclopentyl-6-(2-dimethyl,amino-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-
2-oxo-2,3 -dihydro-1H-indole-5-carbonitrile
Example 120
3-(2-cyclopentyl-6-hydroxy-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-
indole-5-carbonitrile
Example 121
3-{2-cyclopentyl-6-[(2-diethylamino-ethyl)-methyl-amino]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl} -2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Example 122
3-[2-cyclopentyl-6-(2-dimethylamino-ethoxy)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-2-
oxo-2,3-(dihydro-1H-indole-5-carbonitrile
Example 123
3-{2-cyclopentyl-6-[(pyridin-2-yl-methyl)amino]-2H-pyrazolo[3,4-d]pyrimidin-4-yl}-2-
oxo-2,3-dihydro-1H-indole-5-carbonitrile
Example 124
3-(2-cyclopentyl-6-hydrazino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-
1H-indole-5-carbonitrile
Example 125
3-[2-cyclopentyl-6-(2-pyridin-2-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-2-
oxo-2,3-dihydro-1H-indole-5-carbonitrile
Example 126
3-(2-cyclopentyl-6-(2-pyridin-4-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-
oxo-2,3-dihydro-1H-indole-5-carbonitrile
Example 127
3-{2-cyclopentyl-6-[4-(2-pyrrolidin-l-yl-ethyl)-piperazin-1-yl]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl}-2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Example 128
3-{2-cyclopentyl-6-[4-(3-pheny]-propyl)-piperazin-1-yl]-2H-pyrazolo[3,4-d]pyrimidin-
4-yl}-2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Example 129
3-{2-cyclopentyl-6-[4-(2-pyridin-2-yl-ethyl)-piperazin-1-yl]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl}-2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Example 130
3-{2-cyclopentyl-6-[4-(2-thiophen-2-yl-ethyl)-piperazin-1-yl]-2H-pyrazolo[3,4
d]pyrimidin-4-yl}-2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Example 131
3-(2-cyclopentyl-6-piperidin-1-yl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-
dihydro-1H-indole-5-carbonitrile
Example 132
3-(2-cyclopentyl-6-pyrrolidin-1-yl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-
dihyclro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 11.66 (s, 1H), 10.39 (s, 1H), 9.51
(s, 1H), 8.59 (s, 1H), 7.29 (m, 1H), 6.86 (m, 1H), 4.85 (m, 1H), 3.66 (s, 4H), 2.19-1.68
(m, 12H); MS(m/e)414(M +1).
Example 133
3-{2-cyclopentyl-6-[(pyridin-3-ylmemyl)amino]-2H-pyrazolo[3,4-d]pyrimidin-4-yl}-2-
oxo-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.18 (s, 1H), 9.45 (s, 1H), 8.60
(m, 2H), 7.81 (m, 2H), 7.48 (s, 1H), 7.16 (m, 1H), 6.79 (m, 1H), 4.78 (m, 1H), 4.71 (s,
2H), 3.93 (s, 1H), 2.13-1.66 (m, 9H); MS (m/e) 451 (M + 1).
Example 134
3-(2-cyclopentyl-6-2-phenethylamino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-
dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 11.59 (s, 1H), 10.39 (s, 1H), 9.50
(s, 1H), 8.74 (s, 1H), 7.32-7.29 (m, 6H), 6.86 (m, 1H), 4.83 (m, 1H), 3.74 (s, 2H), 2.98
(m, 2H), 2.17-1.67 (m, 9H); MS (m/e) 464 (M + 1).
Example 135
3-{2-Cyclopentyl-6-[4-(1,1-dioxo-l?6-thiomorpholin-4-yl)-phenylamino]-2H-
pyrazolo[3,4-d]pyrimidin-4 -yl} -2-oxci-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.40 (s, 1H), 9.52 (s, 1H), 8.51
(s, 1H), 7.47 (m, 2H), 7.37 (m, 1H), 7.09 (m, 3H), 6.82 (m, 1H), 4.85 (m, 1H), 3.80 (s,
4H), 3.15 (s, 4H), 2.16-1.67 (m, 9H); MS (m/e) 569 (M + 1).
Example 136
3-[2-cyclopentyl-6-(2-pyrrolidin-1-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H 1H NMR (400 MHz, DMSO-d6) d 10.19 (s, 1H), 9.45 (s, 1H),
8.72 (s, 1H), 7.18 (m, 1H), 6.81 (m, 1H), 4.78 (m, 1H), 2.71 (m, 3H), 2.46 ( m, 6H),
2.20-1.60 (m, 13H); MS (m/e) 457 (M +1).
Example 137
3- {2-cyclopentyl-6-[6-(4-methyl-piperazin-1-yl)-pyridin-3-yl-amino]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.31 (s, 1H), 9.65 (s, 1H), 8.32
(m, 1H), 7.61 (m, 1H), 7.25 (m, 1H), 6.95 (m, 1H), 6.86 (m, 1H), 6.73 (m, 1H), 4.94 (m,
1H), 4.64 (m, 3H), 4.06 (m, 2H), 3.90 (m, 2H), 3.55 (s, 3H), 3.20 (m, 2H), 2.18-1.67 (m,
9H); MS (m/e) 535 (M + 1).
Example 138
3-[2-cyclopentyl-6-(3-dimethylamino-propylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.01 (s, 1H), 9.43 (s, 1H), 8.65
(s, 1H), 7.09 (m, 1H), 6.79 (m, O), 4.75 (m, 1H), 3.69 (m, 2H), 3.17 (s, 2H), 2.84 (m,
2H), 2.51 (s, 6H), 2.15-1.62 (m, 1 OH); MS (m/e) 445 (M + 1).
Example 139
3-[2-cyclopentyl-6-(3-morpholin-4-yl-propylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-2-oxo-2,3 -dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.36 (s, 1H), 9.48 (s, 1H), 8.70
(s, 1H), 7.27 (m, 1H), 6.85 (m, 1H), 4.82 (in, 1H), 3.63 (s, 4H), 3.32 (s, 4H), 2.46-2.31
(m, 6H), 2.18-1.67 (m, 10H); MS (m/e) 487 (M + 1).
Example 140
5-Chlorp-3-{2-cyclopentyl-6-[(2-dimethylamino-ethyl)-methyl-amino]-2H-
pyrazolo[3,4-d]pyrimidin-4-yl}-1,3-dihydro-indol-2-one
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 9.90 (s, 1H), 9.51 (s, 1H), 8.28
(s, 1H), 6.83 (m, 1H), 6.69 (m, l.H), 4.81 (m, 1H), 3.81 (s, 2H), 3.26 (s, 3H), 2.69 (s,
2H), 2.31 -1.69 (m, 15H); MS (m/e) 454 (M + 1).
Example 141
5-Chloro-3-[2-cyclopentyl-6-(2-diethylamino-ethoxy)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 11.66 (s, 1H), 9.98 (s, 1H), 9.48
(s, 1H), 8.27 (s, 1H), 6.87 (m, 1H), 6.70 (m, 1H), 4.83 (m, 1H), 3.81 (s, 4H), 2.21-1.65
(m, 14H).
Example 142
5-Chloro-3-[2-cyclopentyl-6-(methylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-1,3-
dihydro-indol-2-one
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 11.54 (s, 1H), 9.97 (s, 1H), 9.48
(s, 1H), 8.44 (s, 1H), 6.86 (m, 1H), 6.70 (m, 1H), 4.80 (m, 1H), 3.00 (s, 3H), 2.13-1.68
(m, 9H); MS (m/e) 383 (M + 1).
Example 143
5-Chloro-3-(2-cyclopentyl-6-morpholin-4-yl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-
dihydro-indol-2-one
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.00 (s, 1H), 9.49 (s, 1H), 8.22
(s, 1H), 6.87 (m, 1H), 6.71 (m, 1H), 4.84 (m, 1H), 3.77 (m, 8H), 2.14-1.69 (m, 9H); MS
(m/e) 439 (M + 1).
Example 144
5-Chloro-3-{2-cyclopentyl-6-[4-(2-dirnethylamino-ethyl)-piperazin-1-yl]-2H-
pyrazolo[3,4-d]pyrimidin-4 -yl}-1,3-dihydro-indol-2-one
Experimental data: '1H NMR (400 MHz, DMSO-d6) d 9.72 (s, 1H), 9.44 (s, 1H), 8.25
(s, 1H), 6.74 (m, 1H), 6.67 (m, 1H), 4.77 (m, 1H), 3.80 (s, 4H), 2.56 (s, 6H), 2.36 (s,
8H), 2.18-1.68 (m, 9H); MS (m/e) 509 (M + 1).
Compound 145
1 -Methyl-1,5-dihydro-pyrazolo[3,4-d]pyrimidin-4-one
A stirred solution of 107 (1 g, 0.0056 mols ) in methanol (5 mL) was stirred overnight at
room temperature. The reaction mixture was concentrated in vacuo, triturated with
ether and collected by filtration to yield a yellow solid (0.8 g, 95% yield). HPLC (82%
purity, retention time 2.52 min.-method F), 1H NMR (400 MHz, DMSO-d6) d 12.15 (s,
1H), 8.07 (d, 1H), 8.04 (s, 1H), 3.80 (s, 3H).
Compound 146
1-Methyl-3-nitro-l,5-dihydro-pyrazolo[3,4-d]pyrimidin-4-one
The nitration to form compound 146 was accomplished following a literature
procedure.3 To a stirred solution of 145 (0.75 g, 0.005 mols) in HNO3 (1.42 d, 6 mL)
and concentrated H2SO4 (12 mL) was heated to 100 °C for 2 h. The mixture was cooled
to room temperature and and poured over ice. The resulting precipitate was collected by
filtration and dried in vacuo to give a yellow solid (0.5 g, 52% yield), m.p. 291-296 °C,
HPLC (86% purity, retention tim; 0.872 min.-method F), 1H NMR (400 MHz, DMSO-
d6) d 12.69 (s, 1H), 8.25 (d, 1H),3.1 (s, 3 H).
Compound 147
4-Chloro-1-methyl -3-nitro-1H-pyrazolo[3,4-d]pyrimidine
Following the procedure for the preparation of compound 25, compound 146 (0.25g,
1.28 mmol) was treated with phosphorous oxychloride (10 mL) and N,N-
dimethylaniline (1 mL). Concentration of the ether afforded 0.270g (98%) of a red solid
which was used without further purification. MS(ES+ calculated - 213.58; found -
214.20 M+H) HPLC (85%) purity, retention time 4.700 minutes-(Method D); 1H NMR
(400 MHz, DMSO-d6) d 9.09 (s, 1H), 4.21 (s, 3H).
Example 148
3-(1-Methyl-3-nitro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-
5-carbonitrile
Utilizing the same procedure for the preparation of Example 29, Example 148 was
prepared with some modifications. To 5-cyanooxindole (0.192g, 1.21mmol) in
anhydrous THF (10 mL) under nitrogen at -78 °C was added lithium diisopropylamine
(Acros, 1.22mL of a 2.0M solution in THF/heptane, 2.43mmoI). The solution was
stirred for fifteen minutes at which point a solution of compound 147 (0.27g, 1.29
mmol) in THF (10 mL) was added dropwise. After addition was complete the external
cooling bath was removed and the reaction was allowed to warm to room temperature.
After 2 hours the reaction was complete. The reaction was quenched by the addition of a
small amount of a saturated ammonium chloride solution and concentrated.
Dichloromethane (5mL) and water (5mL) were added and undissolved solid was
filtered. The solid was washed with minimal dichloromethane, as slight product
solubility was observed. A red solid (0.285g, 70%) was obtained and was used without
further purification: mp>300 C (dec); MS (ES+ calculated - 335.28; found - 336.16
M+H). HPLC (84%) purity; retention time 4.491 minutes-Method D); 1H NMR (400
MHz, DMSO-d6) d 8.39 (s,lH), 7 32 (s, 1H) 6.99 (d, J= 6.24 Hz), 4.03 (s,lH), 4.00 (s,
3H). HPLC Method E:10-100% Acetonitrile over 7 minutes. HPLC purity determined
at 290nm.
The following Examples 149-168 in Table 2 were prepared according to procedures
disclosed herein including the general procedure for the preparation of compounds of
Formula XI, disclosed herein, and using methods generally known to one skilled in the
art.
Example 149
3-(6-Allylamino-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-
1H -indole-5-carbonitrile
A mixture of compound 113 (50 ng, 0.13 mmol), in allylamine (0.99 mL, 1.3 mmol)
and 2-methoxyethanol (5 mL) were heated to 130 °C for 3 h. The reaction was
concentrated, treated with methariol, and filtered. The solid was washed with methanol
and ethyl ether to give 32mg (61%) of the desired product. Example 149: 1HNMR
(400MHz, DMSO-d6) d 11.6 (bs, 1H), 10.4 (s, 1H), 9.5 (s, 1H), 8.7 (s, 1H), 7.3 (d, 1H),
6.8 (d, 1H), 6.0 (m, 1H), 5.0-5.3 (m, 2H), 4.8 (m, 2H), 4.2 (s, 2H), 1.6-2.2 (m, 8H); MS
(m/e) 400 (M + 1); HPLC (99%) purity, retention time 4.212 minutes - Method C; mp
>300°C.
Example 150
3-(2-cyclopentyl-6-isopropylamino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-
dihydro-1H-indole-5-carbonitrile
Example 150 was made in a similar manner to Example 149 using the approriate
starting materials. Experimental c ata: 1H NMR (400MHz, DMSO-d6) d 11.3 (bs, 1H),
10.4 (s, 1H), 9.5 (s, 1H), 8.7 (s, 1H), 7.3 (d, 1H), 6.9 (d, 1H), 4.8 (m, 1H), 4.3 (m, 1H),
3.2 (m, 1H), 1.6-2.2 (m, 8H), 1.3 (d, 6H); MS (m/e) 402 (M + 1); HPLC (98%) purity,
retention time 4.227 minutes - Method C; mp >300°C.
Example 151
3-(6-(2-acetylamino-ethylamino)-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-
oxo-2,3-dihydro-1H-indole-5-carbomtrile
Example 151 was made in a similar manner to Example 149 using the approriate
starting materials. Experimental data: 1H NMR (400MHz, DMSO-d6) d 11.7 (bs, 1H),
10.4 (s, 1H), 9.5 (s, 1H), 8.7 (bs, 1H), 8.0 (m, 1H), 7.3 (d, 1H), 6.8 (d, 1H), 4.8 (m, 1H),
3.6 (m, 2H), 3.4 (m, 3H), 1.6-2.2 (in, 11H); MS (m/e) 445 (M + 1); HPLC (99%) purity,
retention time 3.538 minutes - Method C; mp >300°C.
Example 152
3-(6-Chloro-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
To a solution of Oxindole (260 mg, 1.95 mmol) and THF (5mL) in a 125mL flask at -78
°C was added 2M LDA in THF/Heptane (1.95 mL, 3.9 mmol). After the reaction was
stirred for 30 min, a solution of compound 112 (500mg, 1.95 mmol) in THF (5 mL) was
added and the reaction was continued for 15 min at -78 °C. Next, the reaction was let
come to room temperature and stirred for an additional 2 h. It was quenched with water
(2 mL) and concentrated. The solid was redissolved in methanol and concentrated onto
silica gel. The silica gel was placed onto a column saturated with methylene chloride.
The compound was eluted with a gradient of methylene chloride to 2% methanol /
methylene chloride. The most pure fractions were concentrated, treated with ethyl ether,
and filtered to give 530mg (77%) of the desired product. Example 152: 1HNMR
(400MHz, DMSO-d6) d 11.0-11.2 (bs, 1H), 9.0-9.2 (bs, 1H), 7.7-7.9 (bs, 1H), 6.8-7.2
(m, 4H), 4.8-5.2 (bs, 1H), 1.6-2.3 (m, 8H) ; MS (m/e) 354 (M + 1); HPLC (96%) purity,
retention time 5.206 minutes - Method C; mp 270-273°C.
Example 153
3-(6-Chloro-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-trifluromethyl-1,3-
dihydro-indol-2-one
To a solution of 5-Trifluoromethyloxindole (390 mg, 1.95 mmol) and THF (5mL) in a
125mL flask at -78 °C was added 2M LDA in THF/Heptane (1.95 mL, 3.9 mmol). After
the reaction was stirred for 30 min, a solution of compound 112 (500mg, 1.95 mmol) in
THF (5 mL) was added and the reaction was continued for 15 min at -78 °C. Next, the
reaction was let come to room temperature and stirred for an additional 2 h. It was
quenched with water (2 mL) and concentrated. The solid was redissolved in methanol
and concentrated onto silica gel. The silica gel was placed onto a column saturated with
methylene chloride. The compound was eluted with a gradient of methylene chloride to
2% methanol / methylene chloride. The most pure fractions were concentrated, treated
with ethyl ether, and filtered to give 517mg (63%) of the desired product. Example 153:
1HNMR (400MHz, DMSO-d6) d 10.7 (bs, 1H), 9.5 (bs, 1H), 8.5 (bs, 1H), 7.3 (d,
1H), 7.0 (d, 1H), 4.9-5.0 (m, 1H), 1.6-2.2 (m, 8H); MS (m/e) 422 (M + 1); HPLC (98%)
purity, retention time 5.979 minutes - Method C; mp >300°C.
Example 154
3-(2-cyclopentyl-6-methylamino- 2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-
2-one
A mixture of Compound 152 (40 mg, 0.11 mmol), 2M Methylamine in THF (0.55 mL,
1.1 mmol) and 2-methoxyethanol (2 mL) were heated to 130 °C overnight. The reaction
was concentrated, treated with methanol, and filtered. The solid was washed with
methanol and ethyl ether to give 23mg (61%) of the desired product. Example 154:
lHNMR (400MHz, DMSO-d6) d 13.8 (s, 1H), 10.7 (s, 1H), 8.8 (s, 1H), 7.7 (d, 1H), 7.5
(s, 1H), 6.9-7.1 (m, 2H), 5.0 (m, 1H), 3.0 (m, 1H), 2.8 (s, 3H), 1.6-2.2 (m, 8H); MS
(m/e) 349 (M + 1); HPLC (99%) purity, retention time 3.632 minutes - Method C; mp
>300°C.
Example 155
3-[2-cyclopentyl-6-(2-pyridin-3-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-1,3-
dihydro-indol-2-one
Example 155 was prepared in a similar mariner to Example 154 as disclosed herein
using the approriate starting materials. Experimental data: 1HNMR (400MHz, DMSO-
d6) d 13.8 (s, 1H), 10.7 (s, 1H), 8 8 (s, 1H), 8.4-8.6 (m, 2H), 7.6-7.8 (m, 2H), 7.2-7.5 (m,
1H), 6.9-7.1 (m, 2H), 6.8 (d, 1H) 5.0 (m, 1H), 3.8 (m, 1H), 3.6 (m, 2H), 2.8-3.0 (m,
2H), 1.6-2.2 (m, 8H); MS (m/e) 440 (M + 1); HPLC (90%) purity, retention time 3.376
minutes - Method C; mp 258-260°C.
Example 156
3-{2-cyclopentyl-6-[(pyridin-3-ylmethyl)-amino]-2H-pyrazolo[3,4-d]pyrimidin-4-yl}-
1,3-dihydro-indo1-2-one
Example 156 was prepared in a similar manner to Example 154 as disclosed herein
using the approriate starting materials. Experimental data: 1HNMR (400MHz, DMSO-
d6) d 13.8 (s, 1H), 10.7 (s, 1H), 8.8 (s, 1H), 8.6 (s, 1H), 8.4 (m, 1H), 8.3 (m, 1H), 7.7-7.8
(m, 2H), 7.0 (m, 1H), 6.9 (m, 1H), 5.0 (m, 1H), 4.8 (m, 1H), 4.6 (d, 2H), 1.6-2.2 (m,
8H); MS (m/e) 426 (M + 1); HPLC (97%) purity, retention time 3.476 minutes -
Method C;mp 216-218°C.
Example 157
3-[2-cyclopentyl-6-(3-morpholin-4-yl-propylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one
Example 157 was prepared in a similar manner to Example 154 as disclosed herein
using the approriate starting materials. Experimental data: 1HNMR (400MHz, DMSO-
d6) d 13.8 (s, 1H), 10.7 (s, 1H), 8 8 (s, 1H), 7.7 (m, 2H), 7.0 (m, 2H), 5.0 (m, 1H), 3.6
(m, 4H), 3.5 (m, 1H), 2.4 (m, 6H) 1.6-2.2 (m, 12H); MS (m/e) 462 (M + 1); HPLC
(99%) purity, retention time 3.457 minutes - Method C; mp 242-245°C.
Example 158
3-(2-cyclopentyl-6-(2-acetylamino-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-
dihydro-indol-2-one
Example 158 was prepared in a similar manner to Example 154 as disclosed herein
using the approriate starting materials. Experimental data: lHNMR (400MHz, DMSO-
d6) d 13.8 (s, 1H), 10.7 (s, 1H), 8.8 (s, 1H), 7.7-8.0 (m, 3H), 6.8-7.0 (m, 2H), 5.0 (m,
1H), 3.3 (m, 1H), 3.0 (s, 3H), 1.6-2.2 (m, 12H); MS (m/e) 420 (M + 1); HPLC (95%)
purity, retention time 3.558 minutes - Method C; mp 238-240°C.
Example 159
3(-2-cyclopentyl-6-methylamino-2H-pyra2olo[3,4-d]pyrimidin-4-yl)-5-trifluoromethyl-
1,3-dihydro-indol-2-one
A mixture of Compound 153 (40 mg, 0.095 mmol), 2M Methylamine in THF (0.55 mL,
0.95 mmol) and 2-methoxyethanol (2 mL) were heated to 130 °C overnight. The
reaction was concentrated, treated with methanol, and filtered. The solid was washed
with methanol and ethyl ether to give 25mg (63%) of the desired product. Example
159: 1HNMR (400MHz, DMSO- d6) d 11.6 (s, 1H), 10.3 (s, 1H), 9.5 (s, 1H), 8.8 (s,
1H), 7.2 (d, 1H), 6.9 (m, 2H), 4.8 (m, 1H), 3.0 (d, 3H), 1.6-2.2 (m, 8H); MS (m/e) 417
(M + 1); HPLC (99%) purity, retention time 4.444 minutes - Method C; mp >300°C.
Example 160
3-[2-cyclopentyl-6-(2-pyridin-3-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-
trifluoromethyl-1,3-dihydro-indol-2-one
Example 160 was prepared in a similar manner to Example 153 as disclosed herein
using the approriate starting materials. Experimental data: 1HNMR (400MHz, DMSO-
d6) d 11.4 (s, 1H), 10.3 (s, 1H), 9.5 (s, 1H), 8.7 (s, 1H), 8.4-8.6 (m, 3H), 7.7 (m, 1H),
7.4 (m, 1H), 7.3 (m, 1H), 7.0 (m, 1H), 4.8 (m, 1H), 3.8 (m, 2H), 2.9 (m, 2H) 1.6-2.2 (m,
8H); MS (m/e) 508 (M + 1); HPLC (97%) purity, retention time 3.989 minutes -
Method C;mp278-280°C.
Example 161
3-{2-cyclopentyl-6-[(pyridin-3-ylmethyl)-amino]-2H-pyrazolo[3,4-d]pyrimidin-4-yl}-5-
trifluoromethyl-1,3-dihydro-indol-2-one
Example 161 was prepared in a similar manner to Example 153 as disclosed herein
using the approriate starting materials. Experimental data: 1HNMR (400MHz, DMSO-
d6) d 11.7 (s, 1H), 10.3 (s, 1H), 9.5 (s, 1H), 8.7 (m, 2H), 8.5 (m, 1H), 7.8 (m, 1H), 7.5
(m, 1H), 7.3 (m, 1H), 7.2 (m, 1H), 6.8 (d, 1H), 4.8 (m, 3H), 1.6-2.2 (m, 8H); MS (m/e)
494 (M + 1); HPLC (96%) purity, retention time 4.041 minutes - Method C; mp
>300°C.
Example 162
3-[2-cyclopentyl-6-(3-morpholin-4-yl-propylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-5-trifluoromethyl-1,3-dihydro-indol-2-one
Example 162 was prepared in a similar manner to Example 153 as disclosed herein
using the approriate starting materials. Experimental data: 1HNMR (400MHz, DMSO-
d6) d 11.4 (s, 1H), 10.3 (s, 1H), 9.5 (s, 1H), 8.8 (s, 1H), 7.2 (d, 1H), 7.0 (m, 1H), 6.8 (d,
1H), 4.9 (m, 1H), 3.6 (m, 4H), 2.4 (m, 6H), 1.6-2.2 (m, 12H); MS (m/e) 530 (M + 1);
HPLC (99%) purity, retention time 4.041 minutes - Method C; mp 288-291°C.
Example 163
3-(2-cyclopentyl-6-(2-acetylamino-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-
trifluoromethyl-1,3-dihydro-indol-2-one
Example 163 was prepared in a similar manner to Example 153 as disclosed herein
using the approriate starting materials. Experimental data: 1HNMR (400MHz, DMSO-
d6) d 11.5 (s, 1H), 10.3 (s, 1H), 9.5 (s, 1H), 8.8 (s, 1H), 8.0 (m, 1H), 7.2 (d, 1H), 6.9 (m,
2H), 4.8 (m, 1H), 3.6 (m, 2H), 3.4 (m, 2H), 1.6-2.2 (m, 11H); MS (m/e) 488 (M + 1);
HPLC (99%) purity, retention time 4.314 minutes - Method C; mp >300°C.
Example 164
3-[2-cyclopentyl-6-(2-pyridin-3-y -ethoxy)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-2-oxo-
2,3-dihydro-1H-indole-5-carbonitrile
Example 164 was prepared in a similar manner to Example 153 as disclosed herein
using the approriate starting materials. Experimental data: m.p. 302-304°C; MS
(ES+calculated: 465.52; found: 466 03 M+H). HPLC (98.5% purity, retention time
9.067 minutes - Method B); 1HNMR (400MHz, DMSO-d6) d 10.33 (s, 1H), 9.97 (s,
1H), 9.90 (d, 1H), 9.68 (s,lH), 8.72 (d, 1H), 8,53 (s, 1H), 8.27 (t, 1H), 7.25 (d, 1H), 6.88
(d, 1H), 4.98 (m, 1H), 4.86 (br s, 1H), 3.84 (t, 2H), 3.34 (m, 1H), 3.15 (t, 2H), 2.20 (m,
2H), 2.06 (m, 2H), 1.89 (m, 2H), 1.72 (m, 2H).
Example 165
3-(2-cyclopentyl-6-isobutylamino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-
dihydro-1H-indole-5-carbonitrile
Example 165 was prepared in a similar manner to Example 153 as disclosed herein
using the approriate starting materials. Experimental data: m.p. 297-298.5°C; MS
(ES+calculated: 415.50; found: 415.27 M+H). HPLC (99% purity, retention time
11.175 minutes - Method B); 1HNMR (400MHz, DMSO-d6) d 11.43 (br s.lH), 10.39
(s, 1H), 9.50 (s, 1H), 8.70 (s, 1H), 7.29 (d, 1H), 6.86 (d, 1H), 4.82 (m, 1H), 3.32 (m,
3H), 2.13 (m, 2H), 1.97 (m, 3H), 1.82 (m, 211), 1.68 (m, 2H), 1.00 (d, 6H).
Example 166
3-(2-cyclopentyl-6-ethylamino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-
1H-indole-5 -carbonitrile
Example 166 was prepared in a similar manner to Example 153 as disclosed herein
using the approriate starting materials. Experimental data: m.p. 373°C (dec); MS
(ES+calculated: 387.45; found: 388.22 M+H). HPLC (99% purity, retention time 9.721
minutes-Method B); 1HNMR (400MHz, DMSO-d6) d 11.59 (s, 1H), 10.37 (s, 1H),
9.48 (s, 1H), 8.74 (s, 1H), 7.27 (d, 1H), 6.85 (d, 1H), 4.83 (m, 1H), 3.51 (m, 2H), 3.31
(s, 1H), 2.14 (m, 2H), 1.97 (m, 2HJ, 1.82 (m, 2H), 1.68 (m, 2H), 1.28 (t, 3H).
Example 167
3- {6-[bis-(2-methoxyethyl)amino] - 2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl} -
2-oxo-2,3 -dihydro-1H- indole-5-carbonitrile
Example 167 was prepared in a similar manner to Example 153 as disclosed herein
using the approriate starting materials. Experimental data: m.p. 248.5-249°C; MS
(ES+calculated: 475.55; found: 476 29 M+H). HPLC (99% purity, retention time
11.720 minutes - Method B); 1HNMR (400MHz, DMSO-d6) d 11.64 (s, 1H), 10.42 (s,
1H), 9.52 (s, 1H), 8.42 (s, 1H), 7.29 (d, 1H), 6.87 (d, 1H), 4.87 (m, 1H), 3.93 (s, 4H),
3.65 (m, 4H), 3.61 (m, 1H), 3.32 (s; 6H), 2.16 (m, 2H), 1.98 (m, 2H), 1.84 (m, 2H), 1.70
(m,2H).
Example 168
3-{6-[bis(2-hydroxyethyl)amino]-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl}-2-
oxo-2,3-dihydro-1H-indole-5-carbonitrile
Example 168 was prepared in a similar manner to Example 153 as disclosed herein
using the approriate starting materials. Experimental data: m.p. 243-243.5°C; MS
(ES+calculated: 447.50; found: 448.23 M+H). HPLC (95% purity, retention time 9.084
minutes - Method B); 1HNMR (400MHz, DMSO-d6) d 11.56 (br s, 1H), 10.40 (s, 1H),
9.51 (s, 1H), 8.43 (s, 1H), 7.28 (d, 1H), 6.86 (d, 1H), 5.07 (br s, 2H), 4.84 (m, 1H), 3.87
(s, 4H), 3.78 (m, 4H), 3.70 (m, 1H), 1.97 (m, 2H), 1.83 (m, 2H), 1.69 (m, 2H).
The following Examples 169-182 in Table 3 were prepared according to procedures
disclosed herein using appropriate starting materials and including methods generally
known to one skilled in the art.
Table 3
Example 169
3-[2-Cyclopentyl-6-(pyridin-3-ylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-2-oxo-2,3-
dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.31 (s, 1H), 9.66 (s, 1H), 9.32
(m, 1H), 9.08 (m, 1H), 8.49 (m, 1H), 7.94-7.80 (m, 2H), 7.24-7.22 (m, 1H), 6.88-6.81
(in, 3H), 4.97 (m, 1H), 2.22-1.59 (m, 8H); MS (m/e) 437 (M + 1).
Example 170
3-[2-Cyclopentyl-6-(3-piperidin-1-yl-propylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.27 (s, 1H), 9.46 (s, 1H), 8.73
(s, 1H), 8.03 (s, 1H), 7.22 (s, 1H), 6.82 (m, 1H), 4.81 (m, 1H), 3.46 (m, 2H), 3.08 (m,
2H), 2.63 (m, 2H), 2.43 (m, 2H), 2.15-1.38 (m, 17H); MS (m/e) 485 (M + 1).
Example 171
3-{6-[(6-Chloro-pyridin-3ylmethyl)-amino]-2-cyclopentyl-2H-pyrazolo[3,4-
d]pyrimidin-4-yl}-2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 11.92 (s, 1H), 10.32 (s, 1H), 9.47
(s, 1H), 8.44 (m, 2H), 7.88 (m, 1H), 7.47 (m, 1H), 7.21 (m, 1H), 6.82 (m, 1H), 4.82-4.74
(m, 3H), 2.14-1.66 (m, 9H); MS (m/e) 485 (M + 1).
Example 172
3-[2-Cyclopentyl-6-(methyl-pyri(lin-3-ylmethyl-amino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-2-oxo-2,1-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 11.95 (s, 1H), 10.42 (s, 1H), 9.54
(s, 1H), 8.56 (s, 1H), 8.47 (m, 1H), 8.31 (s, 1H), 7.75 (m, 1H), 7.36 (m, 1H), 7.22 (m,
1H), 6.82 (m, 1H), 5.09 (s, 2H), 4 88 (m, 1H), 3.28 (s, 3H), 2.17-1.68 (m, 8H); MS
(m/e)465(M+l).
Example 173
3-{2-Cyclopentyl-6-[(6-trifluoromethyl-pyridin-3-ylmethyl)-amino]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl} -2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 11.95 (s, 1H), 10.40 (s; 1H), 9.50
(s, 1H), 8.80 (s, 1H), 8.41 (s, 1H), 8.09 (m, 1H), 7.88 (m, 2H), 7.25 (m, 1H), 6.83 (m, 1H), 4.89 (s, 2H), 4.84 (m, 1H), 2.17-1.69 (mi, 8H); MS (m/e) d19 (M + 1).
Example 174
3-[2-Cyclopentyl-6-(2-methoxy-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-2-oxo-
2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 11.53 (s, 1H), 10.39 (s, 1H), 9.49
(s, 1H), 8.66 (s, 1H), 7.29 (m, 1H), 6.86 (m, 1H), 4.83 (m, 1H), 3.65-3.51 (m, 4H); 3.33
(s, 3H), 2.17-1.67 (m, 9H); MS (m/e) 418 (M + 1).
Example 175
3-[2-Cyclopentyl-6-(3-methoxypropylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-2-
oxo-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (40G MHz, DMSO-d6) d 10.14 (s, 1H), 9.44 (s, 1H), 8.71
(s, 1H), 7.16 (m, 1H), 6.80 (m, 1H), 4.77 (m, 1H), 3.48 (m, 4H), 3.32 (m, 2H), 3.23 (s,
3H), 2.14-1.60 (m, 10H); MS (m/s) 432 (M + 1).
Example 176
3-{2-Cyclopentyl-6-[(pyridin-4-ylmethyl)-amino]-2H-pyrazolo[3,4-d]pyrimidin-4-yl}-
2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.27 (s, 1H), 9.47 (s, 1H), 8.55
(m, 2H), 8.48 (m, 2H), 7.40 (m, 2H), 7.19 (m, 1H), 6.79 (m, 1H), 4.80 (m, 1H), 4.74 (s,
2H), 3.94 (s, 1H), 2.14-1.66 (m, 8H); MS (m/e) 451 (M + 1).
Example 177
3-[2-Cyclopentyl-6-(2-morpholin-4-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
2-oxo-2,3-dihydro-1H-indole-5 -carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.37 (s, 1H), 9.48 (s, 1H), 8.70
(s, 1H), 7.27 (m, 1H), 6.85 (m, 1H), 4.83 (m, 1H), 3.68 (m, 4H), 3.59 (m, 2H), 3.31 (m,
4H), 2.65 (m, 2H), 2.53 (m, 2H), 2.16-1.67 (m, 8H); MS (m/e) 473 (M + 1).
Example 178
3-[2-Cyclopentyl-6-(2-thiomorpholin-4-yl-emylammo)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.34 (s, 1H), 9.48 (s, 1H), 8.70
(m, 1H), 7.27 (m, 1H), 6.85 (m, 1H), 4.83 (m, 1H), 3.57 (m, 2H), 2.85-2.55 (m, 12H),
2.16-1.67 (m, 8H); MS (m/e) 489 (M + 1).
Example 179
5-Bromo-3-{2-cyclopentyl-6-[(pyridin-2-ylmethyl)-amino]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl}-1,3-dihydro-1H-indol-2-one
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.82 (s, 1H), 9.97 (s, 1H), 9.49
(s, 1H), 8.57 (m, 1H), 8.44 (m, 1H), 7.79 (m, 1H), 7.50 (m, 1H), 7.43 (m, 1H), 7.30 (m,
1H), 6.97 (m, 1H), 6.65 (m, 1H), 4.85 (m, 2H), 4.80 (m, 1H), 2.15-1.67 (m, 8H); MS
(m/e) d04 (M).
Example 180
5-Bromo-3-{2-cyclopentyl-6-[(pyridin-4-ylmethyl)-amino]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl}-1,3-dihydro-1H-indol-2-one
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 9.88 (s, 1H), 9.48 (s, 1H), 8.49
(m, 2H), 8.28 (s, 1H), 7.39 (m, 3H), 6.91 (m, 1H), 6.62 (m, 1H), 4.77 (m,.3H), 2.15-1.66
(m, 9H); MS (m/e) 504 (M).
Example 181
5-Bromo-3-[2-cyclopentyl-6-(2-pyridin-3-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-
4-yl]-1,3-dihydro-1H-indol-2-one
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 11.37 (s, 1H), 9.98 (s, 1H), 9.49
(s, 1H), 8.62-8.42 (m, 3H), 7.72 (m, 2H), 7.33 (m, 1H), 6.99 (m, 1H), 6.67 (m, 1H), 4.80
(m, 1H), 3.78 (m, 2H), 3.03 (m, 2H), 2.16-1.68 (m, 8H); MS (m/e) d18 (M).
Example 182
5-Bromo-3-(6-chloro-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-
indol-2-one
Experimental data: 1H NMR (400 MHz, DMSO-d6) d 10.48 (s, 1H), 9.34 (s, 1H), 8.26
(s, 1H), 7.17 (m, 1H), 6.78 (m, 1H), 4.98 (m, 1H), 2.20-1.20 (m, 9H); MS (m/e) 433 (M
+ 1).
Example 183
3-(6-Chloro-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-
5-carbonitrile
To a stirring solution of 5-cyanooxindole (103 mg, 0.649 mmol) and anhydrous THF (5
mL) in a 15 mL flask at -78°C was added LDA (649 µL, 1.3 mmol). The reaction
mixture was stirred for 15 min before adding the compound 105 (150 mg, 0.649 mmol)
as a solid. Following an additional 15 min at -78°C, the reaction was allowed to warm
to room temperature and stirred for additional 5 h. The reaction mixture was
concentrated to dryness, taken up into MeOH and concentrated onto silica gel and
pumped dry before subjecting it to flash chromatography on silica gel (gradient elution:
1-10% methanol: dichloromethane) to afford 115 mg (50%) of a yellow solid after
trituration in MeOH. Example 183: mp >300°C; MS (ES+calculated: 352.79; found:
353.29 M+H). HPLC (99%) purity, retention time 11.253 minutes - Method D); 1H
NMR (400 MHz, DMSO-d6) d 10.9 (br s, 1H), 9.37 (s, 1H), 8.41 (bs, 1H), 7.46 (d, J=
8 Hz, 1H), 6.97 (d, J= 8 Hz, 1H), 4.30 (t, J= 7 Hz, 2H), 1.88 (q, J= 3 Hz, 2H), 0.87 (t,
J=7Hz, 3H).
Example 184
2-Oxo-3-{2-propyl-6-[(pyridin-3-ylmethyl)-amino]-2H-pyrazolo[3,4-d]pyrimidin-4-yl}-
2,3-dihydro-1H-mdole-5-carbonitrile
Example 183 (25 mg, 0.071 mmol) and 3-(aminomethyl) pyridine (71.8 µL, 0.71 mmol)
were stirred in 1 mL methoxyethanol overnight at 130°C in an aluminum block. The
reaction mixture was concentrated to dryness and triturated in 3:1 ether/MeOH. The
resulting solid was filtered and pumped dry to afford 23 mg (76%) of a dark yellow
solid. Example 184: mp 291-295°C; MS (ES+calculated: 424.47; found: 425.21 M+H).
HPLC (99%) purity, retention time 8.438 minutes - Method D); 1H NMR (400 MHz,
DMSO-d6) d 11.8 (br s, 1H), 10.39 (s, 1H), 9.43 (s, 1H), 8.64 (s, 1H), 8.46 (d, 1H), 7.83
(d, 1H), 7.37 (dd, J = 5 Hz, J = 3 Hz, 1H), 7.25 (d, J= 8 Hz, 1H), 4.79 (s 2H), 4.16 (t, 7
= 7 Hz, 2H), 1.83 (m, 2H), 1.09 (t, J= 7 Hz, 2H), 0.86 (t, J= 7 Hz, 3H).
Example 185
2-Oxo-3-[2-propyl-6-(2-pyridin-3-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
2,3-dihydro-1H-indole-5-carbonitrile
Using the procedure outlined for Example 184, Example 183 (25 mg, 0.071 mmol) and
3-(aminoethyl) pyridine (86.7 mg, 0.71 mmol) were stirred in 1 mL methoxyethanol
overnight at 130°C in an aluminum block. The reaction mixture was concentrated to
dryness and triturated in a small amount of MeOH. The resulting solid was filtered and
pumped dry to afford 15 mg (48%) of a yellow solid. Example 185: mp 302-305°C;
MS (ES+calculated: 438.50; found: 439.20 M+H). HPLC (99%) purity, retention time
2.996 minutes - Method C); 1H NMR (400 MHz, DMSO-d6) d 11.2 (br s, 1H), 10.40 (s,
1H), 9.43 (s, 1H), 8.71 (s, 1H), 8.51 (s, 1H), 8.42 (d, J= 5 Hz, 1H), 7.74 (d, J = 8 Hz,
1H), 7.33 (m, 2H), 6.86 (d, J= 8 Hz, 1H), 4.16 (t, J= 7 Hz, 2H), 3.77 (br s, 2H), 3.01
(m 2H), 1.84 (m, 2H), 0.86 (t, J= 7 Hz, 3H).
Example 186
2-Oxo-3-{2-propyl-6-[(pyridine-4-ylmethyl)-amino]-2H-pyrazolo[3,4-d]pyrimidin-4-
yl}-2,3-dihydro-1H-indole-5-carbonitrile
Using the procedure outlined for Example 184, Example 183 (25 mg, 0.071 mmol) and
4-(aminomethyl) pyridine (71.8 µL, 0.71 mmol) were stirred in 1 mL methoxyethanol
overnight at 130°C in an aluminum block. The reaction mixture was concentrated to
dryness and triturated in a small amount of MeOH. The resulting solid was filtered and
pumped dry to afford 24 mg (79%) of a yellow solid. Example 186: mp 299-302°C;
MS (ES+calculated: 424.47; found: 425.20 M+H). HPLC (92%) purity, retention time
2.90minutes-MethodC); 1H NMR (400 MHz, DMSO-d6) d 11.87 (brs, 1H), 10.38 (s,
1H), 9.44 (s, 1H), 8.49 (d, 7= 5 Hz, 2H), 8.33 (br s, 1H), 7.40 (d, 7= 5 Hz, 1H), 7.23 (d,
J= 8 Hz, 1H), 6.80 (d, J= 8 Hz, 1H), 6.86 (d, J= 8 Hz, 1H), 4.16 (t, J= 7 Hz, 2H), 3.17
(s, J= 5 Hz, 2H), 1.84 (m, 2H), 0.87 (t, J= 7 Hz, 3H).
Example 187
2-Oxo-3-[2-propyl-6-(2-pyridin-2-yl-emylanuno)-2H-pyrazolo[3,4-d]pyrirnidin-4-yl]-
2,3-clihydro-1H-indole-5-carbonitrile
Using the procedure outlined for Example 184, Example 183 (25 mg, 0.071 mmol) and
2-(aminoethyl) pyridine (84.5 µL, 0.71 mmol) were stirred in 1 mL methoxyethanol
overnight at 130°C in an aluminum block. The reaction mixture was concentrated to
dryness and triturated in a small amount of MeOH. The resulting solid was filtered and
pumped dry to afford 23 mg (74%) of a yellow solid. Example 187: mp >300°C; MS
(ES+calculated: 438.50; found: 439.22 M+H). HPLC (96%) purity, retention time 3.040
minutes - Method C); 1H NMR (400 MHz, DMSO-d6) d 11.55 (br s, 1H), 10.40 (s,
1H), 9.43 (s, 1H), 8.73 (s, 1H), 8.55 (m, 1H), 7.73 (m, 1H) 7.35 (d, J = 8,1H), 7.25 (m,
3H), 6.86 (d, J = 8 Hz, 1H), 4.16 (t, J= 7 Hz, 2H), 3.90 (br s, 2H), 3.16 (m, 2H), 1.84
(m, 2H), 0.86 (t, J= 7 Hz, 3H|.
Example 188
5-Bromo-3-(6-chloro-2-propy1-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-
one
To a stirring solution of 5-bromoc xindole (229 mg, 1.08 mmol) and anhydrous THF (10
mL) in a 25 mL flask at -78°C was added LDA (1.08 mL, 2.16 mmol). The reaction
mixture was stirred for 15 min before adding the substrate 105 (250 mg, 1.08 mmol) as
a solid. Following an additional [5 min at -78°C, the reaction was allowed to warm to
room temperature and stirred overnight (reaction may have been complete after 2 h).
The reaction mixture was concentrated to dryness, taken up into MeOH and
concentrated onto silica gel and pumped dry before subjecting it to flash
chromatography on silica gel (gradient elution: 1-2-3% methanol: dichloromethane) to
afford 210 mg (48%) of a yellow solid that precipitated in the reaction tubes and was
triturated in MeOH. Example 188: mp >300°C; MS (ES+calculated: 406.67; found:
407.87 M+H). HPLC (99%) purity, retention time 12.31 minutes - Method D); 1H
NMR (400 MHz, DMSO-d6) d 10. 5 (br s, 1H), 9.31(s, 1H), 8.26 (br s, 1H), 7.17 (d, J=
8 Hz, 1H), 6.79 (d, J = 8 Hz, 1H), 4.28 (br s, 2H), 1.86 (q, J= 3 Hz, 2H), 0.87 (t, J= 8
Hz, 3H).
Example 189
5-Bromo-3-[2-propyl-6-(2-pyridin-3-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1 3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (25 mg, 0.0614 mmol) and
3-(aminoethyl) pyridine (75 mg, 0.614 mmol) were stirred in 1 mL methoxyethanol
overnight at 130°C in an aluminum block. The reaction mixture was concentrated to
dryness and triturated in a small amount of MeOH. The resulting solid was filtered and
pumped dry to afford 15 mg (50%) of a yellow solid. Example 189: mp 294-297°C;
MS (ES+calculated: 492.38; found: 492.50 M+H). HPLC (80%) purity, retention time
3.437 minutes - Method C); 1H NMR (400 MHz, DMSO-d6) d 11.3 (br s, 1H), 10.01 (s,
1H), 9.42 (s, 1H), 8.52 (m, 2H), S.43 (s, 1H), 7.73 (m, 1H), 7.65 (m, 1H), 7.32 (m, 2H),
6.98 (m, 1H), 6.67 (d, J= 8 Hz) 4.14 (t, J= 7 Hz, 2H), 3.78 (br s, 2H), 3.38 (m), 3.03 (m
2H), 1.84 (m, 2H), 0.86 (m, 3H).
Example 190
5-Bromo-3-{2-propyl-6-[(pyridine-4-ylmethyl)-aimno]-2H-pyrazolo[3,4-d]pyrimidin-4-
yl}-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (25 mg, 0.0614 mmol) and
4-(aminomethyl) pyridine (62 µL, 0.614 mmol) were stirred in 1 mL methoxyethanol
overnight at 130°C in an aluminum block. The reaction mixture was concentrated to
dryness and triturated in a small amount of MeOH. The resulting solid was filtered and
pumped dry to afford 4 mg (14%) of a yellow solid. Example 190: mp >300°C; MS
(ES+calculated: 478.36; found: 478.61 M+H). HPLC (74%) purity, retention time 3.461
minutes - Method C); 1H NMR [400 MHz, DMSO-d6) d 11.69 (br s, 1H), 9.4 (s, 1H),
8.50 (d, J= 6 Hz), 8.24 (s, 1H), 7.42 (d, J= 7 Hz, 1H), 6.93 (d, J = 8 Hz, 1H), 4.79 (d, J
= 6 Hz, 1H), 4.14 (m, 2H), 1.84 (n, 2H), 0.87 (t, J = 7 Hz, 3H).
Example 191
5-Bromo-3-[2-propyl-6-(2-pyridin-2-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (25 mg, 0.0614 mmol) and
2-(aminoethyl) pyridine (73 µL, 0.614 mmol) were stirred in 1 mL methoxyethanol
overnight at 130°C in ah aluminum block. The reaction mixture was concentrated to
dryness and triturated in a small amount of MeOH. The resulting solid was filtered and
pumped dry to afford 14 mg (46%) of a yellow solid. Example 191: mp 283-291 °C;
MS (ES+calculated: 492.38; found: 492.41 M+H). HPLC (86%) purity, retention time
3.60 minutes - Method C); 1HNMR (400 MHz, DMSO-d6) d 11.32 (br s, 1H), 10.02 (s,
1H), 9.42 (s, 1H), 8.55 (d, 2H), 7.75 (m, 1H) 7.37 (d, J= 8,1H), 7.26 (m, 3H), 7.00 (d,
1H), 6.67 (d, J= 8 Hz, 1H), 4.13 (t, J= 7 Hz, 2H), 3.91 (br s, 2H), 3.19 (m, 2H), 1.83
(m, 2H), 0.86 (m, 3H).
Example 192
5-Bromo-3-[2-propyl-6-(3,3,3-trifluoro-propylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one
Example 188 (25 mg, 0.0614 mmol) and 3,3,3-trifluro-n-propylamine hydrochloride.
(89 mg, 0.614 mmol) were stirred in 1 mL methoxyethanol before adding NaH (19.6
mg, 0.491 mmol) and heated o\emight at 130°C in an aluminum block. The reaction
mixture was concentrated to dryness and triturated in a small amount of MeOH. The
resulting solid was filtered and pumped dry to afford 24 mg (81%) of a yellow solid.
Example 192: mp >300°C; MS (ES+calculated: 483.29; found: 483.61 M+H). HPLC
(99%) purity, retention time 4.915 minutes - Method C); 1H NMR (400 MHz, DMSO-
d6) d 11.53 (br s, 1H), 10.86 (s 1H), 10.02 (s, 1H), 9.43 (s, 1H), 8.45 (s, 1H), 7.01 (d,
2H) 6.67 (d, 1H), 4.14 (t, J= 7 Hz, 2H), 3.76 (br s, 2H), 2.70 (m, 2H), 1.83 (m, 2H),
0.86 (m, 3H).
Example 193
3-(6-Allylamino-2-propyl-2H-pyrazolo[3,4-dpyrimidin-4-yl)-5-bromo-1,3-dihydro-
indol-2-one
Example 188 (34 mg, 0.0835 mmol) and allylamine (62.7 µL, 0.835 mmol) were heated
in 1 mL EtOH at 130°C in microwave for 10 min. Upon cooling, the product
precipitated in the reaction tube. The resulting solid was filtered and pumped dry to
afford 30 mg (84%) of a bright yellow solid. Example 193: mp 322-326°C; MS
(ES+calculated: 427.31; found: 428.3 M+H). HPLC (100%) purity, retention time 4.223
minutes - Method C); 1H NMR (400 MHz, DMSO-d6) d 11.37 (s, 1H), 9.98 (s, 1H),
9.41 (s, 1H), 8.52 (s, 1H), 7.11 (br s, 1H), 7.01 (d, 1H), 6.66 (d, J= 8 Hz, 1H), 6.03 (m,
1H), 5.30 (d, 1H), 5.15 (d, 1H) 4.13 (s, 4H), 1.83 (m, 2H), 0.86 (m, 3H).
Example 194
5-Bromo-3-[6-((S)-2-hydroxy-propylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-
yl-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (30 mg, 0.0737 mmol) and
(S)-(+)-1-amino-2-propanol (58 ,xL, 0.737 mmol) were heated in 1 mL EtOH at 130°C
in microwave for 10 min. Upon cooling, the product precipitated in the reaction tube.
The resulting solid was filtered and pumped dry to afford 32 mg (97%) of a bright
yellow solid. Example 194: mp 322-326°C; MS (ES+calculated: 445.32; found: 445.65
M+H). HPLC (97%) purity, retention time 3.735 minutes - Method C); 1H NMR (400
MHz, DMSO-d6) d 11.12 (br s, 1H), 9.99 (s, 1H), 9.41 (s, 1H), 8.52 .(s, 1H), 7.00 (d,
1H), 6.68 (d, 7= 8 Hz, 1H), 4.97 (br s, 1H), 4.13 (t, J= 7 Hz, 2H), 3.94 (br s, 1H), 3.65
(br s, 1H) 3.23 (m, 1H), 1.83 (m, 2H), 1.22 (d, J = 6 Hz, 2H), 1.10 (d, J = 6 Hz, 1H),
0.86 (m, 3H).
Example 195
5-Bromo-3-[6-((R)-2-hydroxy-propylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (30 mg, 0.0737 mmol) and
(R)-(-)-1-amino-2-propanol (58, µL, 0.737 mmol) were heated in 1 mL EtOH at 130°C
in microwave for 10 min. Upon cooling, the product precipitated in the reaction tube.
The resulting solid was filtered and pumped dry to afford 27 mg (82%) of a yellow
solid. Example 195: mp 310°C (dec); MS (ES+calculated: 445.32; found: 445.66
M+H). HPLC (95%) purity, retention time 3.741 minutes - Method C); 1H NMR (400
MHz, DMSO-d6) d 11.12 (br s, 1H), 9.99 (s, 1H), 9.41 (s, 1H), 8.52 (s, 1H), 7.00 (d,
1H), 6.68 (d, J= 8 Hz, 1H), 4.97 (br s, 1H), 4.13 (t, J= 7 Hz, 2H), 3.94 (br s, 1H), 3.65
(br s, 1H) 3.23 (m, 1H), 1.83 (m, 2H), 1.22 (d, J= 6 Hz, 2H), 1.10 (d, J= 6 Hz, 1H),
0.86 (m, 3H).
Example 196
5-Bromo-3-{2-propyl-6-[(pyridin-3-ylmethyl)-amino]-2H-pyrazolo[3,4-d]pyrimidin-4-
yl}-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (30 mg, 0.0737 mmol) and
3-(aminoethyl) pyridine (74.6 µL, 0.737 mmol) were stirred in 1 mL EtOH at 130°C in
microwave for 10 min. Upon cooling, the product precipitated in the reaction tube. The
resulting solid was filtered and pumped dry to afford 15 mg (43%) of a yellow solid.
Example 196: mp 277-281°C; MS (ES calculated: 478.36; found: 478.54 M+H).
HPLC (92%) purity, retention time 3.490 minutes - Method C); 1H NMR (400 MHz,
DMSO-d6) d 9.98 (s, 1H), 9.42 (s 1H), 8.66 (s, 1H), 8.45 (s, 1H), 8.40 (m, 1H), 7.83 (d,
1H), 7.48 (br s, 1H), 7.35 (m, 1H), 6.66 (d, 1H) 4.78 (d, 2H), 4.14 (t, J= 7 Hz, 2H), 1.84
(m, 2H), 0.86 (m, 3H).
Example 197
5-Bromo-3-[6-(2-dimethylamino-ethylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]- 1,3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (30 mg, 0.0737 mmol) and
N,N-dimethylaminoethylamine (65 µL, 0.737 mmol) were heated in 1 mL EtOH at
130°C in microwave for 10 min. Upon cooling, the product precipitated in the reaction
tube. The resulting solid was filtered and pumped dry to afford 16 mg (47%) of a
yellow solid. Example 197: mp 270-274°C; MS (ES+calculated: 458.36; found: 458.69
M+H). HPLC (96%) purity, retiaition time 3.534 minutes - Method C); 1H NMR (400
MHz, DMSO-d6) d 9.97 (s, 1H), 9.41 (s, 1H), 8.51 (s, 1H), 6.99 (d, J= 8 Hz, 1H), 6.85
(br s, 1H), 6.66 (d, J = 8 Hz, 1H), 4.13 (t, J= 7 Hz, 2H), 3.58 (br d, 1H), 2.61 (m, 2H)
2.28 (s, 6H), 1.83 (m, 2H), 0.861 m, 3H).
Example 198
5-Bromo-3-[6-(3-dimethylaminc-propylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-
4-y ]-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (30 mg, 0.0737 mmol) and
N,N-dimethylaminopropylamine (93 µL, 0.737 mmol) were heated in 1 mL EtOH at
130°C in microwave for 10 min. Upon cooling, the product precipitated in the reaction
tube. The resulting solid was filtered and pumped dry to afford 25 mg (79%) of a
yellow solid. Example 198: mp 286-290°C; MS (ES+calculated: 472.39; found: 472.62
M+H). HPLC (98%) purity, retention time 3.464 minutes-Method C); 1H NMR (400
MHz, DMSO-d6) d 9.92 (s, 1H), 9.41 (s, 1H), 8.54 (s, 1H), 7.10 (br s, 1H), 6.97 (d, 7= 8
Hz, 1H), 6.65 (d, J = 8 Hz, 1H), 4.13 (t, J = 7 Hz, 2H), 3.49 (br d, 1H), 2.24 (s, 6H),
1.83 (m, 4H), 0.86 (m, 3H).
Example 199
3-[6-(3-Amino-propylammo)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-bromo-
1,3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (40 mg, 0.0983 mmol) and
1,3-propanediamine (83 µL, 0.983 mmol) were heated in 1 mL EtOH at 130°C in
microwave for 10 min. Upon cooling, the product precipitated in the reaction tube. The
resulting solid was filtered and pumped dry to afford 33 mg (76%) of a yellow solid.
Example 199: mp >300°C; MS (ES+calculated: 444.34; found: 444.61 M+H). HPLC
(94%) purity, retention time 3.297 minutes - Method C); 1H NMR (400 MHz, DMSO-
d6) d 9.50 (s, 1H), 9.37 (s, 1H), 8.57 (s, 113), 7.75 (br s), 6.77 (d, J= 8 Hz, 1H), 6.57 (d,
J= 8 Hz, 1H), 6.35 (br s, 1H) 4.07 (t, J= 7 Hz, 2H), 3.50 (s, 2H), 2.84 (m, 1H), 1.83 (m,
4H), 0.86 (m, 3H).
Example 200
3-[6-(2-Amino-ethylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-bromo-1,3-
dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (40 mg, 0.0983 mmol) and
1,3-ethanediamine (65.7 µL, 0 983 mmol) were heated in 1 mL EtOH at 130°C in
microwave for 10 min. Upon cooling, the product precipitated in the reaction tube. The
resulting solid was filtered and pumped dry to afford 28 mg (66%) of a yellow solid.
Example 200: mp 272-276°C; MS (ES+calculated: 430.31; found: 430.70 M+H).
HPLC (87%) purity, retention time 3.383 minutes - Method C); 1H NMR (400 MHz,
DMSO-d6) d 9.56 (s, 1H), 9.38 (s, 1H), 8.55 (s, 1H), 7.45 (br s), 6.80 (d, J= 8 Hz, 1H),
6.58 (d, J= 8 Hz, 1H), 6.47 (br s, 1H) 4.09 (m, 2H), 3.50 (s, 2H), 3.03 (s, 2H), 1.83 (m,
4H), 0.86 (m, 3H).
Example 201
5-Bromo-3-[6-(3-methylamino-propylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one hydrochloride
Using the procedure outlined for Example 184, Example 188 (50 mg, 0.123 mmol) and
N-(3-aminopropyl)-N-methyl carbamic acid t-butyl ester (231 mg, 1.23 mmol) were
heated in 1 mL EtOH at 130°C in microwave for 10 min. Upon cooling, the product
precipitated in the reaction tube. The resulting solid was filtered and pumped dry before
stirring in 5 mL of 4N HCl/dioxane for 1 h at RT. The reaction mixture was pumped
dry, triturated in ether and filtered to afford 39 mg (64%) of a yellow solid. Example
201: mp 271-273°C; MS (ES+calculated: 458.61; found: 459.4 M+H). HPLC (94%)
purity, retention time 3.406 minutes - Method C); 1H NMR (400 MHz, DMSO-d6) d
8.73 (s, 1H), 7.07 (s, 1H), 6.76 (s, 1H), 4.07 (s, 1H), 3.50 (s, 2H), 3.02 (s, 1H), 2.55 (m,
3H), 2.02 (s, 2H), 1.84 (m, 2H), 0.86 (m, 3H).
Example 202
5-Bromo-3-[6-(2-morpholin-4-yl-eihylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-l 3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (30 mg, 0.0737 mmol) and
N-(2-aminoethyl) morpholine (96 µL, 0.737 mmol) were heated in 1 mL EtOH at
130°C in microwave for 10 min. Upon cooling, the product precipitated in the reaction
tube. The resulting solid was filtered and pumped dry to afford 30 mg (81%) of a yellow
solid. Example 202: mp 299-303°C; MS (ES+calculated: 500.40; found: 500.60 M+H).
HPLC (95%) purity, retention time 3.739 minutes - Method C); 1H NMR (400 MHz,
DMSO-d6) d 9.97 (s, 1H), 9.41 (s, 1H), 8.51 (br s, 1H), 7.00 (d, 1H), 6.87 (m, 1H), 6.66
(d, J = 8 Hz, 1H), 4.13 (t, J= 7 Hz 2H), 3.64 (m, 8H), 2.65 (t, 2H), 1.83 (m, 4H), 0.86
(m, 3H).
Example 203
5-Bromo-3-[6-(3-raorpholin-4-yl-propylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-
4-y}]-l ,3-dihydro-indol-2-one
Using the procedure outlined for Example 184, Example 188 (30 mg, 0.0737 mmol) and
N-(3-aminopropyl) morpholine (107 µL, 0.737 mmol) were heated in 1 mL EtOH at
130°C in microwave for 10 min. Upon cooling, the product precipitated in the reaction
tube. The resulting solid was filtered and pumped dry to afford 30 mg (95%) of a
yellow solid. Example 203: mp 298-303°C; MS (ES+calculated: 514.43; found: 514.51
M+H). HPLC (98%) purity, retention time 3.569 minutes - Method C); 1H NMR (400
MHz, DMSO-d6) d 9.56 (s, 1H), 9.38 (s, 1H), 8.55 (s, 1H), 7.45 (br s), 6.80 (d, J= 8 Hz,
1H), 6.58 (d, J= 8 Hz, 1H), 6.47 (br s, 1H) 4.09 (m, 2H), 3.50 (s, 2H), 3.03 (s, 2H), 2.37
(m, 2H), 1.83 (m, 4H), 0.86 (m, 3H).
Example 204
3-(6-Chloro-2-propyl-2H-pyrazob[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-
5-sulfonic acid dimethylamide
To a stirring solution of 5-dimeihylsulfonamideoxindole (208 mg, 0.866 mmol) and
anhydrous THF (7.5 mL) in a 15 mL flask at -78°C was added LDA (0.866 mL, 1.732
mmol). The reaction mixture was stirred for 15 min before adding the substrate 105
(200 mg, 0.866 mmol) as a solid. Following an additional 15 min at -78°C, the reaction
was allowed to warm to room temperature and stirred over weekend (reaction may have
been complete after 2 h). The reaction mixture was concentrated to dryness, taken up
into MeOH and concentrated onto silica gel and pumped dry before subjecting it to flash
chromatography on silica gel (gradient elution: 1-5% methanol: dichloromethane) to
afford 196 mg (52%) of a yellow solid that precipitated in the reaction tubes and was
triturated in ether. Example 204: mp >300°C; MS (ES+calculated: 434.91; found:
435.42 M+H). HPLC (83%) purity, retention time 4.24 minutes- Method C); 1HNMR
Example 205
3-[6-(2-Dimethylamino-ethylamino)-2-propyl-2H-pyrozolo[3,4-d]pyrimidin-4-yl]-2-
oxo-2,3-dihydro-1H-indole-5-sulfonic acid dimethylamide
Using the procedure outlined for Example 184, Example 188 (50 mg, 0.115 mmol) and
N,N-dimethylaminoethylamine (101 mg, 1.15 mmol) were heated in 1 mL EtOH at
130°C in microwave for 10 min. Upon cooling, the product precipitated in the reaction
tube. The resulting solid was filtered and pumped dry to afford 39 mg (70%) of a
yellow solid. Example 205: mp 259-263°C: MS (ES+calculated: 486.60; found: 487.30
Scheme 4 discloses a general procedure for the preparation of compounds of the
invention wherein R6 is an alkoxyalkyl group and R2 is chloro.
Scheme 4
Compound 206 and 207
5-Amino-1-(2-ethoxyethyl)-1H-pyrazole-4-carbonitrile
3-Amino-1-(2-ethyoxyethyl)-1H-pyrazole-4-carbonitrile
3-Amino-4-cyanopyrazole 1 (3.24g, 30.0 mmol), 2-bromoethoxyethylether (6.12g, 40.0
mmol), and potassium carbonate (5.53ft 40.0 mmol) were combined in 20 mL
anhydrous N,N-dimethylformamide and heated under argon at 80°C overnight. Solids
were removed by filtration and the mother liquor was concentrated to afford after
chromatography on silica (gradient elution 2:1 to 0:1 petroleum ether:ethyl acetate) two
products: KA - a higher RF white solid (2.08g, 39%) and KB - a lower RF pale yellow
solid (2.21g, 41%). Compound 206: rap 130-132°C; MS (ES+calculated: 180.21;
found: 181.16 M+H). HPLC (92% purity, retention time 6.533 minutes - Method A);
*1H NMR (400 MHz, DMSO-d6) d 7.53 (s, 1H), 6.48 (s, 2H), 4.03 (t, J=6Hz, 2H), 3.62
(t, J=6Hz, 2H), 3,42 (q, J=7Hz, 2H), 1.06 (t, J=7Hz, 3H). Compound 207: mp 65-67°C;
MS (ES+calculated: 180.21; found: 181.16 M+H). HPLC (100% purity, retention time
5.277 minutes - Method A); 1H NMR (400 MHz, DMSO-d6): d 8.04 (s, 1H), 5.51 (s,
2H), 4.00 (t, J=6Hz, 2H), 3.64 (t, J=6Hz, 2H), 3.41 (q, J=7Hz, 2H), 1.06 (t, J=7Hz, 3H).
Compound 208
3-Amino-1-(2-ethoxyethyl)-1H-pyrazole-4-carboxylic acid amide
Compound 207 (2.39g, 13.3 mmol) was added in one portion to 3 mL concentrated
sulfuric acid. The mixture was stirred for two hours at which point the mixture had
become homogeneous. The sulfuric acid solution was added dropwise (violent) to
30mL cold concentrated ammoni urn hydroxide solution. The mixture was stirred under
an air stream to dryness over 72 hours. Several milliliters of water were added and a
light brown solid was collected b y filtration. The solid was dried in vacuo to afford
1.977g(75%). Compound 208: VIS (ES+calculated: 198.23; found: 199.80 M+H).
HPLC (73% purity, retention time 2.944 minutes - Method B); 1H NMR (400 MHz,
DMSO-d6): d 7.87 (s, 1H), 7.20 (br s, 1H), 6.67 (br s, 1H), 5.33 (s, 2H), 3.96 (t, J=5Hz,
2H), 3.64 (t, J=5Hz, 2H), 3.39 (q, J=7Hz, 2H), 1.07 (t, J=7Hz, 3H).
Compound 209
2-(2-Ethoxyethyl)-2,7-dihydropyrazolo[3,4-d]pyrimidine-4,6-dione
Compound 208 (1.85g, 0.93 mmol) and urea (5.55g, 92.5 mmol) were mixed and heated
at 200°C to form a melt for two hours. The solution was permitted to cool to room
temperature and 10 mL water wis added. The mixture was refluxed for one hour, was
permitted to cool, and the product was collected by filtration to afford a tan solid
(0.995g, 47%). Compound 209: mp >300°C; MS (ES+calculated: 224.22; found:
224.21 M+). HPLC (75% purit), retention time 3.785 minutes - Method A); 1H NMR
(400 MHz, DMSO-d6): d 9.60 (br s, 2H):, 8.27 (s, 1H), 4.23 (t, J=6Hz, 2H), 3.72 (t,
J=6Hz, 2H), 3.43 (q, J=7Hz, 2H), 1.05 (t, J-7Hz, 3H).
Compound 210
4,6-Dichloro-2-(2-ethoxyethyl)-2H-pyrazolo[3,4-d}pyrimidine
Compound 209 (lg, 4.5 mmol) was suspended in 50 mL phosphorus oxychloride and
was refluxed under argon overnight. The now homogeneous solution was concentrated
in vacuo. Ice was added and the mixture was basified by the additin of 10N sodium
hydroxide solution. The organic s were extracted into ether. The ether was dried
(magnesium sulfate) and was concentrated to afford 0.982g (84%) of a white solid.
Compound 210: MS (ES+calculated: 261.11; found: 261.57 M+). HPLC (95% purity,
retention time 11.686 minutes - Method A); 1H NMR (400 MHz, DMSO-d6): d 9.02 (s,
1H), 4.65 (t, J=5Hz, 2H), 3.90 (t, J=5Hz, 2H), 3.43 (q, J=7Hz, 2H), 1.03 (t, J=7Hz, 3H).
Example 211
5-Chloro-3-[6-chloro-2-(2-ethoxy-ethyl)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-1,3-
dihydro-indol-2-one
To 5-chlorooxindole (168mg, 1.0 mmol) in 5 mL anhydrous tetrahydrofuran under
argon at -78°C was added lithium diisopropylamide (1.05 mL of a 2.0M solution in
THF/hexane, 2.1 mmol) dropwise. The solution was stirred fifteen minutes at which
point Compound 210 (261mg, 1.0 mmol) was added in one portion. The solution was
permitted to warm to room temperature and was stirred for two hours. The solution was
then concentrated and subjected to chromatography on silica (gradient elution 1 to 3%
methanolrdichloromethane). Fractions containing the desired product were further
purified by trituration with methanol to afford following filtration 290mg (74%) of a
yellow solid. Example 211: ntp >300°C; MS (ES+calculated: 392.25; found: 392.61
M+). HPLC (100% purity, retention time 11.993 minutes - Method B); 1H NMR (400
MHz, DMSO-d6): d 10.44 (br s, 1H), 9.30 (br s, 1H), 8.07 (br s, 1H), 7.04 (m, 1H), 6.80
(m, 1H), 4.46 (m, 2H), 3.80 (m, :H), 3.47 (m, 2H), 1.08 (t, J=7Hz, 3H).
Example 212
5-Chloro-3-[2-(2-ethoxy-ethyl)-6-(2-pyridin-3-yl-ethylamino)-2H-pyrazolo[3,4-
d]pyrimidia-4-yl]-1,3-dihydro-indol-2-one
Example 211 (30mg, 0.076 mmol) and 3-(2'-aminoethyl)pyridine (93mg, 0.76 mmol)
were combined in 2 mL ethanol and subjected to reaction in a microwave at 200°C for
ten minutes. On cooling, a brown yellow solid formed which was isolated by filtration.
The solid Was dried in vacuo affording 6mg( 17%). Example 212: mp224-6°C;MS
(ES+calculated: 477.96; found: 478 49 M+H). HPLC (86% purity, retention time 7.996
Example 213
5-Chloro-3-{2-(2-ethoxy-ethyl)-6[(pyridin-3-ylmethyl)-amino]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl}-1,3-dihydro-indol-2-one
Example 211 (30mg, 0.076 mmol) and 3-aminomethylpyridine (82mg, 0.76 mmol) were
combined in 2 mL ethanol and subjected to reaction in a microwave at 130°C for ten
minutes. On cooling a yellow solid formed which was isolated by filtration. The solid
was dried in vacuo affording 22mg (62%). Example 213: mp 294-6°C; MS
(ES+calculated: 463.93; found: 464.44 M+H). HPLC (92% purity, retention time 8.063
Example 214
5-Chloro-3-{2-(2-ethoxy-ethyl)-6-[(pyridine-4-yImethyl)-amino]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl}-1,3-dihydro-indol-2-one
Example 211 was reacted with 4-aminoniethylpyridine to afford a yellow solid. Yield:
45%. Example 214: mp 298-9°0; MS (^calculated: 463.93; found: 464.46 M+H).
HPLC (81% purity, retention time 7.855 minutes - Method B); 1H NMR (400 MHz,
DMSO-d6): d 10.90 (s, 1H), 10.CO (s, 1H), 9.45 (s, 1H), 8.50 (m, 2H), 8.10 (s, 1H), 7.38
(m, 2H), 6.81 (m, 1H), 6.65 (m, 1H), 4.80-4.24 (m, 4H), 3.76 (m, 2H), 3.48 (m, 2H),
1.08 (m,3H).
Example 215
5-Chloro-3-[2-(2-ethoxy-ethyl)-6-(3-methylamino-propylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Example 211 was reacted with N-(3-aminopropyl)-N-methylcarbamic acid-t-butyl ester.
The product obtained by filtration from the ethanolic solution was taken up into 4 mL
4N hydrochloric acid:dioxane and stirred at room temperature for one hour. The
reaction was concentrated and the solid was triturated with ethyl ether to afford after
filtering 28mg (77%) of a yellow solid - isolated as the hydrochloride salt. Example
215: mp 275-7°C; MS (ES+calculated: 443.94; found: 444.46 M+H). HPLC (100%
purity, retention time 7.847 minutes - Method B); 1H NMR (400 MHz, DMSO-d6): d
10.15 (br s, 1H), 9.40 (br s, 1H), 9.83 (m, 2H), 8.34 (br s, 1H), 7.66 (br s, 1H), 7.00 (m,
1H), 6.84 (m, 1H), 4.45 (m, 4H), 3.80 (m, 2H), 3.60-3.45 (m, 5H), 3.04 (m, 2H), 2.56
(m, 2H), 2.02 (m, 2H), 1.08 (m, 3H).
Example 216
5-Chloro-3-[6-(2-dimethylamino-ethylamino)-2-(2-ethoxy-ethyl)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Example 211 was reacted with N,N-dimethylethylenediamine to afford a yellow solid.
Yield: 59%. Example 216: mp 293-5°C; MS (ES+calculated: 443.94; found: 444.49
M+H). HPLC (96% purity, retention time 8.086 minutes - Method B); 1H NMR (400
MHz, DMSO-d6): d 9.97 (s, IB), 9.43 (s, 1H), 8.34 (br s, 1H), 6.88 (m, 2H), 6.69 (d,
J=8Hz, 1H), 4.30 (m, 2H), 3.76 (m, 2H), 3.67 (m, 2H), 3.40 (m, 2H), 3.27 (m, 2H), 2.64
(m, 2H), 2.30 (s, 6H), 1.07 (m, 3H).
Example 217
5-Chloro-3-[2-(2-ethoxy-ethyl)-6-2-morpholin-4-yl-emylamino)-2H-pyrazolo[3,4-
d]pyrimdin-4-yl]-1,3-dihydro-indol-2-one
Example 211 was reacted with N-aminoethylmorpholine to afford a yellow solid. Yield:
62%. Example 217: mp 293-4°C; MS (ES+calculated: 485.98; found: 486.45 M+H).
HPLC (100% purity, retention time 8.513 minutes - Method B); 1H NMR (400 MHz,
DMSO-d6): d 10.84 (s, 1H), 10.00 (s, 1H), 9.47 (s, 1H), 8.38 (br s, 1H), 6.88 (m, 2H),
6.69 (m, 1H), 4.32 (m, 2H), 3.78 (m, 2H), 3.63 (m, 8H), 3.43 (m, 2H), 2.63 (m, 2H),
2.37 (m, 2H), 1.07 (m, 3H).
Example 218
5-Chloro-3-[2-(2-ethoxy-ethyl)-6-((R)-2-hydroxy-propylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Example 211 was reacted with (R)-2-hydroxy-1-aminopropane to afford a yellow solid.
Yield: 95%. Example 218: mp >300°C; MS (ES+calculated: 430.90; found: 431.46
M+H). HPLC (99% purity, retention time 11.949 minutes - Method A); 1H NMR (400
MHz, DMSO-d6): d 11.08 (br s, 1H), 9.95 (s, 1H), 9.42 (s, 1H), 8.32 (s, 1H), 6.85 (m,
1H), 6.68 (m, 1H), 4.96 (br s, IE), 4.30 (m, 2H), 3.78 (m, 2H), 3.67 (br s, 1H), 3.49 (m,
2H), 1.20 (d, J=6Hz, 3H), 1.07 (m, 3H).
Example 219
5-Chloro-3-[2-(2-ethoxy-ethyl)-6((S)-2-hydroxy-propylamino)-2H-pyrazolo[3,4-
d]pyrimicin-4-yl]-1,3-dihydro-indol-2-one
Example 211 was reacted with (S)-2-hydroxy-1-aminopropane to afford a yellow solid.
Yield: 89%. Example 219: mp >300°C; MS (ES+calculated: 430.90; found: 431.43
M+H). HPLC (99% purity, retention time 11.924 minutes - Method A); 1H NMR (400
MHz, DMSO-d6): d 11.10 (br s, 1H), 9.95 (s, 1H), 9.43 (s, 1H), 8.32 (s, 1H), 6.82 (m,
1H), 6.66 (m, 1H), 4.96 (br s, IB), 4.30 (m, 2H), 3.78 (m, 2H), 3.67 (br s, 1H), 3.43 (m,
2H), 1.20 (d, J=6Hz, 3H), 1.07 (m, 3H).
Scheme 5 discloses a general procedure for the preparation of compounds of the
invention wherein R6 is an alkoxyalkyl group and R2 is bromo.
Scheme 5
Compound 220 and 221
5-Amino-1-(2-methoxyethyl)-1H-pyrazole-4-carbonitrile
3-Amino-1-(2-methoxyethyl) -1H-pyrazole-4-carbonitrile
3-Amino-4-cyanopyrazole (1) was reacted wiith bromomethylmethyl ether to afford LA
(43%) and LB (49%) as white solids. Compound 220: mp 120-122°C; MS
(ES+calculated: 166.18; found: 167.26 M+H). HPLC (90% purity, retention time 4.716
minutes - Method A); 1H NMR (400 MHz, DMSO-d6): d 7.52 (s, 1H), 6.49 (br s, 2H),
Compound 222
3-Amino-1-(2-methoxyethyl)-1H-pyrazole-4-carboxylic acid amide
Compound 221 was reacted with sulfuric acid to afford LC as a white solid (100%).
Compound 222: MS (ES+calculated: 184.20; found: 185.64 M+H). HPLC (92% purity,
retention time 2.138 minutes - Method A); 1H NMR (400 MHz, DMSO-d6): d 7.87 (s,
1H), 7.20 (br s, 1H), 6.69 (br s, 1H), 5.33 (s, 2H), 3.97 (t, J=5Hz, 2H), 3.60 (t, J=5Hz,
2H), 3.22 (s, 3H).
Compound 223
2-(2-Methoxyethyl)-2,7-dihydropyrazolo[3,4-d]pyrimidine-4,6-dione
Compound 222 was reacted with urea to afford a white solid. Yield: 81%. Compound
223: mp 295-302°C; MS (ES+calculated: 210.19; found: 211.20 M+). HPLC (90%
purity, retention time 2.833 minutes - Method B); 1H NMR (400 MHz, DMSO-d6): d
11.06 (br s, 1H), 10.64 (br s, 1H), 8.28 (s, 1H), 4.25 (t, J=5Hz, 2H), 3.69 (t, J=5Hz, 2H),
3.23 (s, 3H).
Compound 224
4,6-Dichloro-2-(2-methoxyethyl)-2H-pyrazolo[3,4-d]pyrimidine
Compound 223 was reacted witf i phosphorus oxychloride to afford a white solid. Yield:
83%. Compound 224: MS (ES" calculated: 247.09; found: 247.97 M+). HPLC (94%
purity, retention time 7.685 minutes - Method B); 1H NMR (400 MHz, DMSO-d6): d
9.03 (s, 1H), 4.66 (t, J=5Hz, 2H), 3.86 (t, J=5Hz, 2H), 3.24 (s, 3H).
Example 225
5-Bromo-3-[6-chloro-2-(2-methoxy-ethyl)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-1,3-
dihydro- indol-2-one
5-bromooxindole was condensed with Compound 224 to afford an orange solid. Yield:
93%. Example 225: mp >300°C MS (ES+calculated: 422.67; found: 423.90 M+H).
HPLC (96% purity, retention time 11.474 minutes - Method B); 1H NMR (400 MHz,
DMSO-d6): d 10.48 (br s, 1H), 9.26 (br s, 1H), 8.26 (br s, 1H), 7.16 (m, 1H), 6.72 (m,
1H), 4.46 (m, 2H), 3.77 (m, 2H), 3.27 (s, 3H).
Example 226
5-Bromo-3-[2-(2-methoxy-ethyl)-6-(3-methylamino-propylamino)-2H-pyrazolo[3,4-
d]pyrimdin-4-yl]-1,3-dihydro-indol-2-one
Example 225 was reacted with N-(3-aminoptropyl)-N-methylcarbamic acid-t-butyl ester.
The product obtained by filtration from the ethanolic solution was taken up into 4 mL
4N hydrochloric acid:dioxane and stirred at room temperature for one hour. The
reaction was concentrated and this solid was triturated with ethyl ether to afford after
filtering 28mg (77%) of a yellow solid - isolated as the hydrochloride salt. Example
226: mp 232-6°C; MS (ES+calculated: 474.36; found: 475.88 M+H). HPLC (85%
Example 227
5-Bromo-3-[2-(2-methoxy-ethyl)-6-(2-piperidin-4-yl-ethylamino)-2H-pyrazolo[3J4-
djpyrimidin-4-yl]-1,3-dihydro-indol-2-one
Example 225 was reacted with 4-aminoethyl-1-N-BOC piperidine to afford after
removal of the BOC group a yellow solid - the hydrochloride salt. Yield: 74%.
Example 227: mp 248-51°C; MS (ES+calcuIated: 514.43; found: 515.92 M+H). HPLC
Example 228
5-Bromo-3-[2-(2-methoxy-ethyll-6-(2-piperidin-3-yl-ethylamino)-2H-pyrazolo[3,4-
d]pyrirmdin -4-yl]-l ,3-dihydro-indol-2-one
Example 225 was reacted with 3-aminoethyl-1-N-BOC piperidine to afford after
removal of the BOC group a yellow solid - the hydrochloride salt. Yield: 59%.
Example 228: mp 270-3°C; MS (ES+calculated: 514.43; found: 515.92 M+H). HPLC
(100% purity, retention time 7.674 minutes - Method B); 1H NMR (400 MHz, DMSO-
Example 229
5-Bromo-3-[6-(2-dimethylamino-ethylamino)-2-(2-methoxy-ethy])-2H-pyrazolo[3,4-
d]pyrimdm-4-yl]-1,3-dihydro-indol-2-one
Example 225 was reacted with N,N-dimethylethylenediamine to afford a yellow solid.
Yield: 65%. Example 229: mp 291-2°C; MS (ES+calculated: 474.36; found: 475.84
M+H). HPLC (95% purity, retention time 7.827 minutes - Method B); 1H NMR (400
MHz, DMSO-d6): d 10.80 (br s, 1H), 9.95 (br s, 1H), 9.38 (s, 1H), 8.47 (br s, 1H), 6.98
(d, J=8Hz, 1H), 6.64 (d, J=8Hz, HI), 4.30 (m, 2H), 3.72 (m, 2H), 3.56 (m, 2H), 3.26 (s,
3H), 2.60 (m, 2H), 2.24 (s, 6H).
Example 230
5-Bromo-3-[6-((S)-2-hydroxy-propylamino)-2-(2-methoxy-ethyl)-2H-pyrazolo[3,4-
d]pyrimidin- 4-yl]-1,3-dihydro-indol-2-one
Example 225 was reacted with (S)-2 -hydroxy- 1-aminopropane to afford a yellow solid.
Yield: 82%. Example 230: mp >300°C; MS (ES+calcuIated: 461.32; found: 462.79
M+H). HPLC (94% purity, retentioia time 8.225 minutes - Method B); 1H NMR (400
MHz, DMSO-d6): d 11.16 (br s, 1H), 10.00 (s„ 1H), 9.39 (s, 1H), 8.50 (s, 1H), 6.98 (d,
J=8Hz, 1H), 6.63 (d, J=8Hz, 1H), 4.97 (br s, 1H), 4.41 (m, 1H), 4.30 (m, 2H), 3.90 (br s,
1H), 3.76 (m, 1H), 3.68 (m, 2H), 3.23 (s, 3H), 1.20 (d, J=8Hz, 3H).
Example 231
5-Bromo-3-[6-((R)-2-hydroxy-propylamino)-2-(2-methoxy-ethyl)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Example 225 was reacted with (R)-2-hydroxy-1-arninopropane to afford a yellow solid.
Yield: 64%. Example 231: mp >300°C; MS (ES+calculated: 461.32; found: 462.79
M+H). HPLC (98% purity, retention time 8.242 minutes - Method B); 1H NMR (400
MHz, DMSO-d6): d 11.10 (br s, 1H), 9.96 (s, 1H), 9.40 (s, 1H), 8.48 (s, 1H), 6.98 (d,
J=8Hz, 1H), 6.63 (d, J=8Hz, 1H), 4.97 (br s, 1H), 4.46 (m, 1H), 4.31 (m, 2H), 3.92 (br s,
1H), 3.77 (m, 1H), 3.72 (m, 2H), 3.24 (s, 3H), 1.19 (d, J=8Hz, 3H).
Example 232
5-Bromo-3-{2-(2-methoxy-ethyl)-6-[(pyridin-3-ylmethyl)-amino]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl}-1,3-dihydro-indol-2-one
Example 225 was reacted with 3-aminomethylpyridine to afford a yellow solid. Yield:
65%. Example 232: mp 300-301°C; MS (ES+calculated: 494.35; found: 495.82 M+H).
HPLC (89% purity, retention time 7.618 minutes - Method B); 1H NMR (400 MHz,
DMSO-d6): d 10.87 (s, 1H), 1001 (s, 1H), 9.44 (s, 1H), 8.70-8.40 (m, 4H), 7.37 (m,
1H), 6.96 (m, 1H), 6.60 (m, 1H), 4.80 (m, 2H), 4.34 (m, 2H), 3.77 (m, 2H), 3.24 (s, 3H).
Example 233
5-Bromo-3-[6-(3-dimethylamino-propylainino)-2-(2-methoxy-ethyl)-2H-pyrazolo[3,4-
d1pyrimklin-4-yl]-1,3-dihydro-indol-2-one
Compound 225 was reacted with N,N-dimethylpropylenediamine to afford a yellow
solid. Yield: 58%. Example 233: mp 297-8°C; MS (ES+calculated: 488.39; found:
489.89 M+H). HPLC (94% purity, retention time 7.519 minutes - Method B); 1H NMR
(400 MHz, DMSO-d6): d 10.80 (s, 1H), 9.90 (s, 1H), 9.40 (s, 1H), 8.52 (s, 1H), 67.0 (d,
J=8Hz, 2H), 6.68 (d, J=8Hz, 1H), 4.30 (m, 2H), 3.74 (m, 2H), 3.50 (m, 2H), 3.35 (m,
2H), 3.21 (s, 3H), 2.27 (s, 6H), 1.32 (m, 2H).
Example 234
5-Bromo-3-[2-(2-methoxy-ethyl)-6-(2-morpholin-4-yl-ethylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-l ,3 -dihydro-indol-2-one
Example 225 was reacted with N-aminoethylmorpholine to afford a yellow solid. Yield:
55%. Example 234: mp >300°C; MS (ES+calculated: 516.40; found: 517.82 M+H).
HPLC (97% purity, retention time 8.150 minutes - Method B); 1H NMR (400 MHz,
DMSO-d6): d 10.82 (s, 1H), 10.00 (s, 1H), 9.42 (s, 1H), 8.48 (br s, 1H), 7.02 (m, 2H),
6.62 (m, 1H), 4.32 (m, 2H), 3.72 (m, 2H), 3.63 (m, 8H), 3.31 (m, 5H), 2.70 (m, 2H).
Example 235
5-Bromo-3-[2-(2-methoxy-emyl)-6-(2-methoxy-ethylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Example 225 was reacted with 2-methoxyethylamine to afford a yellow solid. Yield:
100%. Example 235: mp >300ºC; MS (ES+calculated: 461.32; found: 462.77 M+H).
HPLC (97% purity, retention time 8.518 minutes - Method B); 1H NMR (400 MHz,
DMSO-d6): d 11.20 (br s, 1H), 10.00 (s, 1H), 9.40 (s, 1H), 8.46 (s, 1H), 6.96 (d, J=8Hz,
2H), 6.64 (d, J=8Hz, 1H), 4.30 (m, 2H), 3.70 (m, 2H), 3.68 (m, 2H), 3.50 (m, 2H), 3.29
(s, 3H), 3.22 (s, 3H).
Example 236
5-Bromo-3-[6-chloro-2-(2-ethoxy-ethyl)-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-
dihydro-indol-2-one
To a stirring solution of 5-bromooxindole (163 mg, 0.769 mmol) and anhydrous THF (7
mL) in a 15 mL flask at -78°C was added LDA (0.769 mL, 1.538 mmol). The reaction
mixture was stirred for 15 min before adding the Compound 210 (200 mg, 0.769 mmol)
as a solid. Following an additional 15 min at -78°C, the reaction was allowed to warm
to room temperature and stirred overnight. The reaction mixture was concentrated to
dryness, taken up into MeOH and concentrated onto silica gel and pumped dry before
subjecting it to flash chromatography on silica gel (gradient elution: 1-10% methanol:
dichloromethane) to afford 228 mg (68%) of a yellow solid after concentrated and was
triturated in ether. Example 236: mp 260-263°C; MS (ES+calculated: 436.70; found:
437.92 M+H). HPLC (91%) purity, retention time 4.946 minutes - Method C); 1H
NMR (400 MHz, DMSO-d6) d 10.50 (br s, 1H), 9.3(br s, 1H), 8.3 (br s, 1H), 7.17 (d, J
Example 237
5-Bromo-3-[6-(2-dimethylamino-ethylamino)-2-(2-ethoxy-ethyl)2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 212, Example 236 (30 mg, 0.0687 mmol) and
N,N-dimethylaminoethylamine (60.5 µL, 0.687 mmol) were heated in 1 mL EtOH at
130°C in microwave for 10 miia. Upon cooling, the product precipitated in the reaction
tube. The resulting solid was filtered and pumped dry to afford 24 mg (71%) of a
yellow solid. Example 237: nip 247-251°C; MS (ES+calculated: 488.39; found: 489.4
M+H). HPLC (94%) purity, retention time 3.410 minutes-Method C); 1HNMR(400
MHz, DMSO-d6) d 9.99 (s, 1H), 9.44 (s, 1H), 8.63 (s, 1H), 7.02 (d, 1H), 6.67 (d, 1H),
4.31 (t, 2H), 3.76 (m, 2H), 3.61 (br s, 1H), 3.42 (m),'2.33 (s, 2H) 2.18 (s, 1H), 0.86 (m,
9H).
Example 238
5-Bromo-3-[2-(2-ethoxy-ethyl)-6-((S)-2-hydroxy-propylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 212, Example 236 (30 mg, 0.0687 mmol) and
(S)-(+)-1-amino-2-propanol (54 µL, 0.687 mmol) were heated in 1 mL EtOH at 130°C
in microwave for 10 min. Upon cooling, the product precipitated in the reaction tube.
The resulting solid was filtered and pumped dry to afford 26 mg (80%) of a yellow
solid. Example 238: mp 297-299°C; MS (ES+calculated: 475.35; found: 475.2 M+H).
HPLC (94%) purity, retention time 2.785 minutes - Method C); 1H NMR (400 MHz,
Example 239
5-Bromo-3-[2-(2-ethoxy-ethyl)-6-((R)-2-hydroxy-propylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 212, Example 236 (30 mg, 0.0687 mmol) and
(R)-(-)-1-amino-2-propanol (54 µL, 0.687 mmol) were heated in 1 mL EtOH at 130°C
in microwave for 10 min. Upon cooling, the product precipitated in the reaction tube.
The resulting solid was filtered aitd pumped dry to afford 28 mg (86%) of a yellow
solid. Example 239: mp 298-300' C; MS (ES+calculated: 475.35; found: 475.2 M+H).
Example 240
5-Bromo-3-[2-(2-ethoxy-ethyl)-6-(2-morpholin-4-yl-ethylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Using the procedure outlined for Example 212, Example 236 (30 mg, 0.0737 mmol) and
N-(2-aminoethyl) morpholine (90 µL, 0.737 mmol) were heated in 1 mL EtOH at 130°C
in microwave for 10 min. Upon cooling, the product precipitated in the reaction tube.
The resulting solid was filtered and pumped dry to afford 29 mg (80%) of a yellow
solid. Example 240: mp 276-278°C; MS (ES+calculated: 530.43; found: 530.61 M+H).
HPLC (96%) purity, retention time 2.757 minutes - Method C); 1H NMR (400 MHz,
DMSO-d6) ): d 10.00 (s, 1H), 9.45 (s, 1H), 8.53 (s, 1H), 7.00 (d, 1H), 6.87 (m, 1H), 6.66
The following Examples 241-254 in Table 4 were prepared according to procedures
disclosed herein including using methods generally known to one skilled in the art.
Table 4
Example 241
3-{2-Cyclopentyl-6-[(pyridin-4-ylmethyl)-amino]-2H-pyrazolo[3,4-d]pyrimidin-4-yl}-
1,3-dihydro-indol-2-one
A mixture of Example 152 (40 mg, 0.11 mmol), in 4-aminomethylpyridine (119mg, 1.1
mmol) and 2-methoxyethanol (2 mL) were heated to 130 °C for 5 h. The reaction was
concentrated. The Example was purified via a flash silica gel column eluting with 5% to
10% methanol / methylene chloride. The most pure fractions were concentrated, treated
with ethyl ether and filtered to give 23mg (50%) of Example 241. Example 241:
1HNMR (400MHz, DMSO-d6) d 13.9 (s, 1H), 10.7 (s, 1H), 8.6 (m, 2H), 8.4 (m, 1H), 7.8
(m, 1H), 7.3 (m, 2H), 7.0 (m, 2H), 6.9 (m, 1H), 5.0 (m, 1H), 4.8 (m, 1H), 4.6 (d, 2H),
1.6-2.2 (m, 8H); MS (m/e) 426 (M + 1); HPLC (95%) purity, retention time 3.472
minutes - Method C; mp 198-200°C.
Example 242
3-[2-Cyclopentyl-6-(2-pyridin-2 yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-1,3-
dihydro-indol-2-one
Examples 242 was synthesized i i a similar manner to Example 241 as disclosed herein
using the approriate starting materials. Example 242: 1HNMR (400MHzj DMSO-d6)
d 13.7 (s, 1H), 10.7 (s, 1H), 8.8 is, 1H), 8.6 (m, 1H), 7.8 (m, 3H), 7.3 (m, 1H), 7.2 (m,
1H), 7.0 (m, 1H), 5.8 (s, 2H), 5.0 (m, 1H), 3.7 (m, 2H), 3.1 (m, 2H), 1.6-2.2 (m, 8H);
MS (m/e) 440 (M + 1); HPLC (99%) purity, retention time 3.397 minutes - Method C;
mp 246-248°C.
Example 243
3-(6-Allylamino-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-
one
Examples 243 was synthesized in a similar manner to Example 241 as disclosed herein
using the approriate starting materials. Example 243: 1HNMR (400MHz, DMSO-d6)
d 13.8 (s, 1H), 10.7 (s, 1H), 8.8 (s, 1H), 7.9 (m, 1H), 7.7 (m, 1H), 6.7-7.0 (m, 4H), 5.9-
6.1 (m, 1H), 5.0-5.3 (m, 2H), 3.8 (m, 2H), 1.6-2.2 (m, 8H); MS (m/e) 375 (M + 1);
HPLC (99%) purity, retention time 4.082 minutes - Method C; mp >300°C.
Example 244
3-(2-Cyclopentyl-6-isobutylamino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-
indol-2-one
Examples 244 was synthesized in a similar manner to Example 241 as disclosed herein
using the approriate starting materials. Example 244: 1HNMR (400MHz, DMSO-d6)
Example 245
3-(2-Cyclopentyl-6-isopropy[amino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-
indol-2-one
Examples 245 was synthesized in a similar manner to Example 241 as disclosed herein
using the approriate starting materials. Example 245: 'jHNMR (400MHz, DMS0-d6)
d 13.6 (s, 1H), 10.7 (s, 1H), 8.7 (s, 1H), 7.7 (d, 1H), 7.5 (d, 1H), 6.7-7.1 (m, 3H), 5.0 (m,
1H), 4.1 (m, 1H); 1.6-2.2 (m, ?H), 1.2 (m, 6H); MS (m/e) 377 (M + 1); HPLC (92%)
purity, retention time 4.124 mir.utes - Method C; mp 303-305°C.
Example 246
3-{2-Cyclopentyl-6-[(pyridin-4-ylmethyl)-amino]-2H-pyrazolo[3,4-d]pyrimidin-4-yl}-
5-trifluoromethyl-1,3-dihydro-indol-2-one
A mixture of Example 153 (40 mg, 0.095 mmol), in 4-Arninomethylpyridine (103mg,
0.95 mmol) and 2-methoxyethaiiol (2 mL) were heated to 130 °C for 5 h. The reaction
was concentrated. The Example was purified via a flash silica gel column eluting with
5% methanol / methylene chloride. The most pure fractions were concentrated, treated
with ethyl ether and filtered to give 6mg (13%) of Example 246. Example 246:
1HNMR (400MHz, DMSO-d6) d 11.7 (s, 1H), 10.3 (s, 1H), 9.5 (s, 1H), 8.5 (m, 3H), 7.5
(m, 1H), 7.3 (m, 2H), 7.1 (m, IB), 6.8 (d, 1H), 4.8 (m, 3H), 1.6-2.2 (m, 8H); MS (m/e)
494 (M + 1); HPLC (94%) purity, retention time 3.979 minutes - Method C; mp
>300°C.
Example 247
3-[2-Cyclopentyl-6-(2-pyridin-2-yl-emylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-
trifluoromethyl-1,3-dihydro-indol-2-one
508 (M + 1); HPLC (97%) parity, retention time 4.140 minutes - Method C; mp
>300°C.
Example 248
3-(6-Allylamino-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-trifluoromethyl-
1,3-dihydro-indol-2-one
Example 248 was synthesized in a similar manner to Example 246 as disclosed herein
Example 249
3-(2-Cyclopentyl-6-isobutylamino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-
trifluoromethyl-1,3-dihydro-indol-2-one
Example 249 was synthesized in a similar manner to Example 246 as disclosed herein
Example 250
3-(2-Cyclopentyl-6-isopropylamino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-
trifluoromethyl-1,3-dihydro-indol-2-one
Example 251
3-[2-Cyclopentyl-6-(2-dimethylamino-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
5-trifluoromethyl-1,3-dihydro-indol-2-one
Example 251 was synthesized ia a similar manner to Example 246 as disclosed herein
using the approriate starting materials. Example 251: 1HNMR (400MHz, DMSO-d6) d
10.3 (s, 1H), 9.5 (s, 1H), 8.7 (s, LH), 7.2 (d, 1H), 6.9 (d, 1H), 4.8 (m, 1H), 3.4 (m, 2H),
2.9 (t, 2H), 2.4 (t, 2H), 2.2 (s, «3H), 1.6-2.2 (m, 8H); MS (m/e) 474 (M + 1); HPLC
(91%) purity, retention time 4.076 minutes - Method C; mp 211-213°C.
Example 252
3-[6-(2-Amino-emylamino)-2-cyclopentyl-2H-pyrazolo[3)4-d]pyrimidin-4-yl]-5-
trifluoromethyl-l,:i-dihydroindol-2-one hydrochloride salt
A mixture of Example 153 (63mg, 0.15mmol), N-BOC-ethylenediamine (0.24gm,
1.5mmol), and ethanol (lmL) wers heated in a microwave at 130°C for 10 minutes. The
reaction was cooled to room temperature and a precipitate formed. The solid was
filtered, washed with ethanol and ethyl ether to give 56mg (68%) of the intermediate.
Next, the material was stirred in 4N HC1 in 1,4-dioxane (2mL). After 1 hour the reaction
was concentrated. The solid was treated with ether and filtered to give 35mg (70%) of
Example 252. Example 252: lHNMR (400MHz, DMSO-dg) d 12.0 (s, 1H), 10.3 (s, 1H), 9.5 (s, 1H), 8.6 (s, 1H), 8.0 (m, 2H), 7.4 (m, 2H), 6.9 (m, 1H), 4.8 (m, 1H), 3.7 (m,
2H), 3.2 (m, 2H), 1.6-2.2 (m, 8H); MS (m/e) 446 (M + 1); HPLC (99%) purity, retention
time 3.917 minutes - Method C; mp 290-293°C.
Example 253
3-[2-Cyclopentyl-6-(3-methylamino-propylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
5-trifluoromethyl-1,3 -dihydro-indol-2-one hydrochloride salt
Example 253 was synthesized in a similar manner to Example 252 as disclosed herein
using the approriate starting materials. Example 253: 1HNMR (400MHz, DMSO-d6)
d 11.8 (bs, 1H), 10.3 (bs, 1H), 9.5 (bs, 1H), 8.8 (s, 1H), 8.7 (m, 1H), 7.4 (m, 1H), 7.2
(m, 1H), 6.9 (m, 1H), 4.8 (m, 1H), :i.6 (m, 2H), 3.0 (m, 4H), 1.6-2.2 (m, 11H); MS (m/e)
475 (M + 1); HPLC (99%) purity, retention time 3.977 minutes - Method C; mp 260-
262°C.
Example 254
3-[6-(3-Amino-propylamino)-2-cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-
trifluoromethyl-1,3-dihydro-indol-2-one hydrochloride salt
Example 254 was synthesized in a similar manner to Example 252 as disclosed herein
using the approriate starting materials. Example 254: 1HNMR (400MHz, DMSO-d6)
d 11.9 (bs, 1H), 10.3 (bs, 1H), 9.5 (bs, 1H), 8.8 (s, 1H), 8.1(m, 2H), 7.7 (m, 1H), 7.3
(m, 1H), 6.9 (m, 1H), 4.8 (m, 1H), 3.6 (m, 2H), 3.0 (m, 4H), 1.6-2.2 (m, 8H); MS (m/e)
460 (M + 1); HPLC (97%) purity, retention time 3.850 minutes - Method C; mp 304-
306°C.
Compound 255 and 256
5-Amino-1-isobutyl-1H-pyrazole-4-carbonitrile
3-Amino-1-isobutyl-1H-pyrazole-4-carbonitrile
3-Amino-4-pyrazolecarbonitrili; 1 (Aldrich, 24.0g, 0.222 mole), l-bromo-2-methyl-
propane (Lancaster, 36.48g, 0.266 mole) and anhydrous potassium carbonate (Acros,
36.8g, 0.266 mole) were suspended in 240 mL reagent grade acetonitrile and heated at
80°C under nitrogen for 22 hours. LC/MS indicated 1 remained. Therefore, added an
additional 3 mL (0.027 mole) 1-bromo-2-methylpropane and 4.25g (0.031 mole) K2CO3
After 24 hours, the reaction was filtered and the filtrate concentrated in vacuo. The
solid was stirred in 250 mL of water for 3.5 hours at room temp. The solid was filtered,
washed with 50 mL thethyl ethisr, and dried to yield 16.28g (45%) of a tan powder
which was seen by 1H NMR to contain a mixture of 255 and 256 in approximately a 2:1
ratio. This mixture was used without further purification. Mixture of 255 and 256: mp
91.3-106; MS (ES+calculated: 164.21; found: 165.28 M+H). HPLC (99% purity,
retention times 6.920 and 7.086 minutes - Method B); 1H NMR (400 MHz, DMSO-d6) d
8.05 and 7.51 (s, 1H), 6.50 and 5.49 (s, 2H), 3.68 (t, 2H), 2.05 (m, 1H), 0.82 and 0.81
(d,6H).
Compound 257 and 258
5-Amino-1-isobutyl-1H-pyrazole-4-carboxylic acid amide
3-Amino-1-isobutyl-1H-pyrazole-4-carboxylic acid amide
To concentrated sulfuric acid (Fisher, 48 mL) at 0°C was added 229 and 230 (5.89g,
36.0 mmol) in small portions. The reaction was allowed to warm to room temperature
and was stirred over 4.25 hours. The viscous reaction was added slowly (violent) to 240
mL concentrated ammonium hydroxide solution (Fisher) over 25 minutes. Added
300mL of water to the mixture and extracted with EtOAc, dried over MgSCu, filtered,
and concentrated in vacuo to afford 5.48g (84%) of an off-white solid which was seen
by 1H NMR to contain a mixtuie of 257 amd 258 in approximately a 2:1 ratio. Mixture
of 257 and 258: mp 154-157.8ºC; MS (Eg calculated: 182.23; found: 183.23 M+H).
HPLC (99% purity, retention time 6.498 minutes - Method B); 1H NMR (400 MHz,
DMSO-d6) d 7.84 and 7.61 (s, 1H), 7.10 (br s, 1H), 6.70 (br s, 1H), 6.14 and 5.31 (s,
1H), 3.64 (m, 2H), 2.05 (m, 1H) 0.83 (d, 6H).
Compound 259
2-Isobutyl-2H-pyrazolo[3,4-d]pyrimidine-4,6-diol
Heated 1.575g (8.64 mmol) and 4.725g (78.7 mmol) urea at 200°C in a selade tube for
3.5 hours in a sealed tube. The mixture was cooled to 100°C, and 34 mL water was
added. The mixture was refluxed at 100°C for 20 hours. The reaction was cooled to
room temperature, additional water was added, and the product was extrated using ethyl
acetate. The organic layer was dried with MgSCM, filtered, and cone, in vacuo to give
989 mg (55%) of a white solid which was seen by 1H NMR to contain only 259: mp
327.5-330°C; MS (ES+calculated: 208.22; found: 209.25 M+H). HPLC (99% purity,
retention time 5.631 minutes - Method B); 1H NMR (400 MHz, DMSO-d6) d 11.32 (s,
1H), 10.64 (s, 1H), 8.32 (s, 1H), 3.90 (d, 2H), 2.12 (m, 1H), 0.84 (d, 6H).
Compound 260
4,6-Dichloro-2-isobutyl-2H-pyrazolo[3,4-d]pyrimidine
Compound 259 (500mg, 2.4 mraol) was dissolved in phosphorus oxychloride (Acros, 8
mL) and the mixture was refhxed under argon at 110°C for 29.5 hours. Excess
phosphorus oxychloride was removed in vacuo and ice then water was added to the dark
orange syrup. Next, 10N NaOH was added until the pH equaled 14. The product was
then extracted with methylene chloride, dried over MgSO4, filtered and cone, in vacuo.
afford a white solid which was purified by flash chromatography on silica gel eluting
with 500:11 dichloromethane:methanol to afford 388mg (25%) of a white solid.
Compound 260: MS (ES+calcuhted: 245.11; found: 245.40 M). HPLC (99%) purity,
retention time 10.794 minutes - Method B); 1H NMR (400 MHz, DMSO-d6) d 9.05 (s,
1H), 4.34 (d, 2H), 2.33 (m, 1H), C .90 (d, 6H).
Example 261
5-Chloro-3-(6-chloro-2-isobutyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-
2-one
To 5-cyanooxindole (CombiBlocks, 238mg, 1.42 mmol) in 7.0 mL anhydrous THF
under argon at -78°C was added lithium diisopropylamine (Acros, 1.42 mL of a 2.0M
solution in THF/heptane, 2.84 mmol). The solution was stirred for fifteen minutes at
which point compound 260 (348mg, 1.42 mmol) was added. The reaction was stirred
for fifteen minutes, external cooling was removed, and the reaction was allowed to
warm to room temperature. After 2.75 hours the solution was quenched with water.
The reaction was concentrated in vacuo and the residue was purified by flash
chromatography on silica gel eluting with 99:1 dichloromethane:methanol to afford 125
mg of Example 261 as an orange solid: mp 284°C (dec); MS (ES+calculated: 376.25;
found: 376.62 M). HPLC (95%) purity, retention time 13.170 minutes - Method B); lH
Example 262
5-Bromo-3-(6-chloro-2-isobu tyl-2H-pyiazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-
2-one
To 5-bromooxindole (CombiBlocks, 216mg, 1.02 mmol) in 7.0 mL anhydrous THF
under argon at -78°C was added lithium diisopropylamine (Acros, 1.02 mL of a 2.0M
solution in THF/heptane, 2.04 mmol) over 4 minutes. The solution was stirred for thirty
minutes at which point a solution of compound 260 (250mg, 1.02 mmol) in 3.13 mL dry
THF was added over 3 minutes. The reaction was stirred for twenty-three minutes, the
external cooling was removed, and the reaction was permitted to warm to room
temperature. After 3 hours the solution was quenched with water. The reaction was
concentrated in vacuo and the residue was purified by flash chromatography on silica
gel eluting with 98:2 then 95:5 dichloromethane:methanol to afford Example 262 as a
burnt orange solid in two separate lots totaling 183 mg (43%): m.p. 295°C (decomp.);
MS (ES+calculated: 420.70; found: 421.94 M+H). HPLC (89% purity, retention time
Example 263
5-Bromo-3-[2-isobutyl-6-(2-methoxy-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
1,3-dihydro-indol-2-one
Example 264
5-Bromo-3-[2-isobutyl-6-(2-morpholin-4-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-
4-y1]-1,3-dihydro-indol-2-one
Example 265
5-Bromo-3-[2-isobutyl-6-(3morpholin-4-yl-propylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Example 267
5-Bromo-3-[2-isobutyl-6-(3-methylamino-propylamino)-2H-pyrazolo[3,4-d]pyrimidin-
4-yl]-l ,3-dihydro-indol-2-one
Example 268
5-Bromo-3-[6-(2-dimethylamino-ethylamino)-2-isobutyl-2H-pyrazolo[3,4-d]pyrimidin-
4-yl]-l,3-dihydro-indol-2-one
Example 269
5-Bromo-3-[6-(3-dimethylamino-propylamino)-2-isobutyl-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Example 270
5-Chloro-3-[6-(2-dimethylamino-ethylamino)-2-isobutyl-2H-pyrazolo[3,4-d]pyrimidin-
4-yl]-1,3-dihydro-indol-2-one
Example 271
5-Chloro-3-[2-isobutyl-6-(3-morpholin-4-yl-propylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-l ,3-dihydro-indol-2-one
Example 272
5-Chloro-3-[2-isobutyl-6-(2-methoxy-ethyhimino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
1,3-dihydro-indol-2-one
Example 273
5-Chloro-3-(6-chloro-2-isobutyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-
2-one
Example 275
3- {6-[(1-Butyl-piperidin-4-yImethyl)-amino]-2-cyclopentyl-2H-pyrazolo[3,4-
d]pyrimidin-4-yl}-5-chloro-1,3-dihydro-indol-2-one
Example 276
N-{2-[4(5-Chloro-2-oxo-2,3-dihydro-1H-indol-3-yl)-2-cyclopentyl-2H-pyrazolo[3,4-
d]pyrimidin-6-ylamino]-ethyl}-acetamide
Example 277
5-Chloro-3-[2-cyclopentyl-6-(2-methyoxy-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-4,3-dihydro-indol-2-one
Example 278
5-Chloro-3-[2-cyclopentyl-6-(3-morpholin-4-yl-propylamino)-2H-pyrazolo[3,4-
d]pyrimidin 4-yl]-l ,3-dihydro-indole-2-one
Example 279
5-Chloro-3-[2-cyclopentyl-6-(2-pyridin-3-yl-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-
4-yl] -1,3-dihydro-indol-2-one
Example 280
5-Chloro-3-(2-cyclopentyl-6-isobutylamino-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-
dihydro-indol-2-one
Compound 281 and 282
3-Amino-1-pentyl-1H-pyrazole-4-carbonitri le
5-Amino-1-pentyl-1H-pyrazole-4-carbonitrile
3-Amino-4-pyrazolecarbonitrile 1 (Acros, 15 g, 0.138 mol), pentyliodide (Acros, 24.45
ml, 0.187 mol) and anhydrous potassium carbonate (Fisher, 25.88 g, 0.187 mol) were
suspended in 100 mL anhydrous DMF and heated at 80°C under nitrogen overnight.
HPLC analysis (Method D) showed starting material still present. An additional 9 ml
(69 mmol) of petnyliodide and 9.5 g (69 mrnol) K2CO3 were added to the reaction and
heating continued for 5 more hours. The reaction was permitted to cool and the DMF
was removed on a rotary evaporator. Water was added (100 mL) and the organics were
extracted with dichloromethane (3 X 100 mL). The combined dichloromethane
fractions were washed with water (50 mL) and brine (50 mL) and were dried
(magnesium sulfate). Concentration of the organics afforded orange solid which
contained both pentyl isomers and residual DMF by NMR analysis (27.65 g, greater
than theoretical yield). This was carried on crude. HPLC (1 peak, 63 %) elutes at 9.5
minutes (Method D). 1H NMR (400 MHz, DMSO-d6) d 7.87 (s, 1H), 7.60 (s, 1H), 6.15
(s, 1H), 5.33 (s, 2H), 3.80 (t, J=7,2H), 1.68 (m, 2H), 1.25 (m, 4H) 0.75 (t, 7=7,3H).
Compound 283 and 284
3-Amino-1-pentyl-1H-pyrazole-4-caroboxylic acid amide
5-Amino-1-pentyl-1H-pyrazole-4-carboxylic acid amide
To 4 ml cone. H2SO4 at 0° C, was added 1.6 g (8.9 mmol) of the crude mixture
(Example 254 and Compound 255) from the above alkylation reaction. The reaction
was allowed to stir and warm to room temperature. After 4 h stirring, the entire solid
had been dissolved. The thick, acid solution was added dropwise to stirring, ice cold
NH4OH (aq). The resulting white precipitate (1.23 g, 70 %), as a mixture of the two
isomers, was collected via vacuum filtration, washed with water and dried in vacuo. 1H
NMR (400 MHz, DMSO-d6) d 7.82 (s, 1H), 7.58 (s, 1H), 7.15 (br s, 2H), 6.65 (br s,
2H), 5.32 (s, 1H), 3.80 (t, 7=7 Hz, 2H), 1.68 (m, 2H), 1.25 (m, 4H), 0.75 (t, 7=7 Hz,
3H).
Compound 285
2-Pentyl-2Hpyrazole[3,4-d]pyrimidine-4,6-diol
In a sealed tube 539 mg (2.75 mmol) of the mixture of isomers (Compound 283 and
Compound 284) from the above reaction and 1.65 g (27.5 mmol) urea were combined
and heated to 180°C. The solids became an off white liquid and then returned to solid at
approximately 3 h of heating. The reaction was cooled to 130° C and 10 ml water
added. The aqueous solution was refluxed overnight, cooled to room temperature in the
morning, and filtered. Only the desired isomer was collected as a white solid (177 mg,
29%). HPLC (87 %) elutes at 7.8 3 min. (Method B). 1H NMR (400 MHz, DMSO-d6) d
11.35 (s, 1H), 11.65 (s, 1H), 8.35 (s, 1H), 4.10 (t, J=7 Hz, 2H), 1.80 (m, 2H), 1.25 (m,
4H),0.75(t,J=7Hz,3H).
Compound 286
4,6-Dichloro-2-pentyl-2H-pyrazolo[3,4-d]pyrimidine
To 30 ml POCI3, was added 1.12g compound 285 prepared as above. The solution was
refluxed for 5 h and monitored by removing aliquots, quenching them into saturated
NaHC03 and extracting with ether. The POCI3 was first removed via rotary evaporation
and subsequent high vacuum for approximately 1 h. The resulting dark syrup was
quenched into stirring ice water and the aqueous solution made basic with 5 % NaOH.
The solution was transferred to a separatory funnel and extracted with 3 x 50 ml
portions of ether. The combined ether was dried over MgSO4, filtered and evaporated to
a white to off white solid 900 me, 69%). HPLC (99 %) elutes at 11.78 min. (Method
Example 287
3-(6-Chloro-2-pentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-trifluoromethyl-1,3-dihydro-
indol-2-one
To a stirring solution of 5-trifluoromethyloxindole (300 mg, 1.49 mmol) and THF (5
mL), in 50 mL flask at -78°C was added LDA (1.86 mL, 3.73 mmol). After the reaction
was stirred for 45 min, a solution of compound 286 prepared above (402 mg, 1.55
mmol) in THF (3 mL x 2) was added and continued to stir for 1 h at -78°C. The
reaction was allowed to warm to loom temperature and stirred for additional 2 h. It was
quenched with saturated NH4Cl (10 mL). The resulting precipitate was filtered, washed
with water, and dried under house vacuum at 50°C overnight to give 587 mg (93%) of
General Procedure for Aminations using Microwave -
In a CEM 10 ml disposable microwave tube equipped with a stir bar, was placed 75 mg
(0.18 mmol) of the oxidolepyrazolopyrimidine prepared above and 2 ml EtOH. To the
tube was added 10 eq. of the correfjpqnding amine and the tube heated to 130°C for 10
min in the microwave reactor. The resulting solid was filtered and washed with ether
and dried in vacuo. If no solid was formed, the EtOH was evaporated and the resulting
solid triturated with ether, filtered and dried in vacuo.
Example 288
3-[6-(2-Amino-ethylamino)-2-pentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-
trifluoromethyl-1,3-dihydro-indol-2-one
Example 289
3-[6-(3-Amino-propylamino)-2-pentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-
trifluorom ethyl-1,3-dihydro-indol-2-one
Example 290
3-[6-(3-Morpholin-4-yl-propylamino)-2-pentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-
trifluorometltyl-1,3-dihydro-indol-2-one
Example 291
3-(6-Chloro-2-pentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-
5-carbonitrile
To a stirring solution of 5-cyanoloxindole (166 mg, 1.05 mmol) and THF (2 mL) in 50
mL flask at -78°C was added IDA (1.31 mL, 2.62 mmol). After the reaction was stirred
for 45 min, a solution of compound 286 prepared above (290 mg, 1.11 mmol) in THF (1
mL x 2) was added and continued to stir for 1 h at -78°C. The reaction was allowed to
warm to room temperature and stirred for additional 2 h. It was quenched with saturated
NH4CI (10 mL). The resulting precipitate was filtered, washed with water, and dried
under house vacuum at 50°C overnight to give 115 mg (29%) of the desired product as a
General Procedure for Aminations in Radley's tube -
In a Radley's Tube equipped with a stir bar, was placed 50 mg (0.18 mmol) of the 5-
cyano oxidolepyrazolopyrimidine prepared above and 2 ml 2-methoxyethanol. To the
tube was added 10 eq. of the corresponding amine and the tube heated to 130°C and the
reaction followed by LC/MS. When reaction was complete, the solvent was removed
via Speedi-Vac overnight. The resulting solid suspended in MeOH, filtered, washed
with ether and dried in vacuo.
Example 292
2-Oxo-3-{2-pentyl-6-[(pyridin-4-ylmethyl)-amino]-2H-pyrazolo[3,4-d]pyrimidin-4-yl}-
2,3-dihydro-1H-indole-5-carbonitrile
Example 293
3-(6-Allylamino-2-pentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-
indole-5-carbonitrile
Example 294
3-(6-Methylamino-2-pentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-
indole-5-carbonitrile
Example 295
3-(6-Isopropylamino-2-pentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-
1H-indole-5-carbonitrile
Scheme 6
Compound 296
3-Amino-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester
Methylhydrazine (5.90g, 128 mmo ) and p-ardsaldehyde (17.43g, 128 mmol) were
refluxed in 100 mL dry benzene employing a Dean-Start trap to remove water. After 24
hours the organics were concentrated and the reaction was reconstituted by the addition
of 50 mL anhydrous benzene. Ethyl ethoxymethylenecyanoacetate (21.65g, 128 mmol)
in SO mL anhydrous benzene was added dropwise and the mixture was refluxed for one
hour. The reaction was concentrated and the remaining organics were triturated with
ethanol to afford a solid after filtering. To this solid was added approximately 100 mL
ethanol and 17 mL concentrated hydrochloric acid. The mixture was stirred !4 hour at
80°C at which point the reaction became homogeneous. The reaction was concentrated
and the product obtained was triturated with atpproximately 500 mL boiling ethyl ether
for one hour to remove anisaldehyde. The suspended solid was filtered off and
dissolved in chloroform (approximately 250 mL). The chloroform solution was washed
with saturarated sodium bicarbonate solution and was dried (magnesium sulfate) and
concentrated to afford 19.66g (91%) of a peach solid Compound 296 which was used in
succeeding steps without purification.
Compound 297
2-Methyl-2,7-dihydropyrazolo[3,4-d]pyrimidine-4,6-dione
Compound 296 (2g, 11.8 mmoll was melted at 200°C with 6 g urea (large excess) for
two hours. The reaction was permitted to cool to 40°C and 20 mL water was added.
The mixture was then boiled foi 1 hour and stirred at room temperature overnight.
Filtration and drying in vacuo afforded 1.86 g (95%) of a white solid. Compound 297:
mp>300°C;MS(ES+calculated 166.14; found: 167.24 M+H). HPLC (100% purity,
retention time 2.103 minutes - Method A):; 1H NMR (400 MHz, DMSO-d6): d 11.34 (br
s, 1H), 10.66 (br s, 1H), 8.27 (s, 1H), 3.82 (s, 3H).
Compound 298
2-Methyl-6-thioxo-2,5,6,7-tetrahydropyrazolo[3,4-d]pyrimidin-4-one
Compound 296 (2g, 11.8 mmol) was melted at 200°C with 6 g thiourea (large excess)
for two hours. The reaction was permitted to cool to 40°C and 50 mL water was added.
The mixture was then boiled overnight. Filtration and drying in vacuo afforded 1.06 g
(49%) of a white solid. Compound 298: mp >300°C; MS (ES+calculated: 182.20;
found: 183.18 M+H). HPLC (100% purity, retention time 5.328 minutes - Method D);
1H NMR (400 MHz, DMSO-d6): d 13.05 (br s, 1H), 12.86 (br s, 1H), 8.70 (s, 1H), 3.88
(s, 3H).
Compound 299
4,6-Dichloro-2-methyl-2H-pyrazolo[3,4-d]pyrimidine
Compound 297 (1.04g, 5.7 mmol) was refluxed under argon with 50 mL phosphorous
oxychloride for 24 hours. Following introduction of a pipette for analysis, the
suspension became homogeneous after approximately an hour. Following homogeneity
the reaction was refluxed an additional hoar. Excess phosphorus oxychloride was then
scrupulously removed in vacuo (high vacuum). Ice was added and the mixture
neutralized to basic by addition of ION sodium hydroxide solution. A yellow solid was
removed by filtration and dried in vacuo to afford 0.76g (66%). Compound 299: MS
(ES+calculated: 203.03; found: 203.19 M+H). HPLC (67% purity-pdt decomposes on
hplc column, retention time 6.895 minutes - Method A); 1H NMR (400 MHz, DMSO-
d6): d 9.02 (s, 1H), 4.23 (s, 3H).
Example 300
5-Bromo-3-(6-chloro-2-methyl-2H-pyrazo lo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-
one
To 5-bromooxindole (212mg, 1.0 mmol) in 5 mL anhydrous tetrahydrofuran at -78°C
under argon was added lithium di isopropylamide (1.05mL of a 2.0M solution in
THF/hexane, 2.1 mmol) dropwise. The solution was stirred ten minutes and Compound
299 (202mg, 1.0 mmol) was added in one portion. The reaction was permitted to warm
to room temperature at which point 2 mL of N-methylpyrrolidinone was added to
promote homogeneity. Dissolution occurred and the reaction was stirred for 90 minutes,
The reaction was concentrated and residual solvent was removed via lyophilization.
The resulting organics were applied to silica gel and eluted (gradient from 1 to 10%
methanohdichloromethane) to afford 275mg (73%) of an orange solid. Example 300:
mp >300°C; MS (ES+calculated: 378.62; found: 380.02 M+H). HPLC (78% purity,
retention time 11.002 minutes - Method B); 1H NMR (400 MHz, DMSO-d6): d 10.47
Example 301
3-(6-Allylamino-2-methyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-bromo-1,3-dihydro-
indol-2-one
Example 300 (30mg, 0.08 mmol), allylamine (46mg, 0.90 mmol) and methoxyethanol
(2mL) were combined and heated in a sealed tube at 130°C overnight. The reaction was
concentrated to afford a solid which was triturated with 1 mL methanol. Filtration and
drying afforded 19 mg (60%) of a yellow solid. Example. 301: mp 336-340°C; MS
(ES+calculated: 399.25; found: 400.77 M+H). HPLC (96% purity, retention time 9.036
Example 302
5-Bromo-3-{2-methyl-6-[(pyridin-4-ylmethyl)-amino]-2H-pyrazolo[3,4-d]pyrimidin-4-
yl}- l,3-dihydro-indol-2-one
Example 302 was prepared as a gum from the reaction of Example 300 and 4-
aminomethylpyridine. Yield: 12mg (33%). Example 302: MS (ES+calculated: 450.30;
found: 451.79 M+H). HPLC (93% purity, retention time 7.220 minutes - Method B);
Example 303
3-[6-(2-Amino-ethylamino)-2-methyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-bromo-1,3-
dihydro-indol-2-one
Example 303 was prepared by reacting Example 300 and ethylenediamine. Yield: 19
mg (59%). Example 303: mp 260-2°C; MS (ES+calculated: 402.26; found: 403.79
M+H). HPLC (86% purity, retention time 7.036 minutes - Method B); 1H NMR (400
MHz, DMSO-d6): d 9.56 (br s, 111), 9.37 (br s, 1H), 8.55 (br s, 1H), 6.79 (m, 1H), 6.57
(m, 1H), 6.43 (br s, 1H), 3.88 (s, 3H), 3.60-3.20 (m, 7H).
Example 304
5-Bromo-3~[6-(2-hydroxypropy]amino)-2-methyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
1.3-dihydro-indol-2-one
Example 304 was prepared from the reaction of Example 300 and (S)-2-hydroxy-1-
aminopropane. Yield: 16 mg (48%). Example 304: mp 338-40°C; MS (ES+calculated:
417.27; found: 418.72 M+H). HP1-C (97% purity, retention time 7.743 minutes -
Method B); lH NMR (400 MHz, DMSO-d6): d 10.00 (s, 1H), 9.37 (s, 1H), 8.50 (s, 1H),
7.00 (d, J=8Hz, 1H), 6.67 (d, J=8He, 1H), 3.91 (s, 3H), 3.70-3.20 (m, 4H), 1.22 (m, 3H).
Example 305
5-Bromo-3-[2-methyl-6-(2-pyridin-3-yl-emylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one
Example 300 (30mg, 0.08 mmol) and 3-(2:aminoethyl)pyridine (98mg, 0.80 mmol) in 2
mL ethanol was subjected to reaction at 130°C in a microwave for ten minutes. On
cooling a mustard brown solid was collected by filtration. It was washed with ethanol
and dried in vacuo to afford 24n\g (65%). Example 305: mp 314-7°C; MS
(ES+calculated: 464.33; found: 465.78 M+H). HPLC (96% purity, retention time 7.201
minutes - Method B); 1H NMR (400 MHz, DMSO-d6): d 9.89 (s, 1H), 9.36 (s, 1H),
8.60-8.40 (m, 3H), 7.72 (m, 1H), 7.32 (m, 1H), 6.94 (d, J=8Hz, 1H), 6.80 (br s, 1H),
6.64 (d, J=8Hz, 1H), 3.90 (s, 3H), 3.85-2.80 (m, 5H).
Example 306
5-Bromo-3-[2-methyl-6-(3-methylamino-propylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one
In a similar fashion to Example 305, BOC protected Example 306 was prepared from
the reaction of Example 300 and N-(3-aminopropyl)-N-methylcarbamic acid-t-butyl
ester. The product obtained by filtration from the ethanolic solution was taken up into 4
mL 4N hydrochloric acidrdioxane and stirred at room temperature for one hour. The
reaction was concentrated and the solid was triturated with ethyl ether to afford after
filtering 17mg (46%) of a yellow solid - isolated as the hydrochloride salt, mp 271-
273°C; MS (ES+calculated: 430.31; found: 431.91 M+H). HPLC (92% purity, retention
time 8.228 minutes - Method D); 1H NMR (400 MHz, DMSO-d6): d 10.10 (br s, 1H),
9.34 (br s, 1H), 8.64 (br s, 1H), 7.27 (br s, 1H), 7.04 (m, 1H), 6.72 (m, 1H), 3.95 (s, 3H),
3.38 (d, J=7Hz, 3H), 3.78-1.80 (m, 9H).
Example 307
5-Bromo-3-[6-(3-dimethylamino-propylamino)-2-methyl-2H-pyrazolo[3,4-d]pyrimidin-
4-yl]-1,3-dihydro-indol-2-one
In a similar manner as for the preparation Example 305, Example 307 was prepared
from the reaction of 300 and N,N-dimethylpropylenediamine in ethanol to afford 10mg
(28%) of a yellow solid. Example 307: mp 310-12°C (dec); MS (ES+calculated:
444.34; found: 445.88 M+H). HPLC (97% purity, retention time 7.155 minutes -
Example 308
3-(6-Chloro-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-methyl-1,3-dihydroindol-2-
one
A solution of 5-methyloxindole (147 mg, 1.0 mmol) and compound 105 (231 mg, 1.0
mmol) in 2 mL dry THF and 2 mL dry DMF was cooled to 0 ºC, and a dispersion of
60% NaH in mineral oil (80 mg, 2.0 mmol) was added. After the hydrogen evolution
ceased, the reaction mixture was allowed to warm to rt. The reaction mixture was stirred
under N2 for 2 days. The dark yellow homogeneous reaction was quenched with sat.
NH4C1, and a yellow precipitate formed. The precipitate was washed with water, and the
product allowed to air dry. The crude product was then suspended in thethyl ether, and
the yellow solid recovered by filtration affording g (80%) of the product: mp 275-80 ºC.
Example 309
3-[6-(2-Dimethylamino-ethylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-
methyl-1,3-dihydro-indol-2-one
Using microwave procedure as described previously, the homogeneous reaction mixture
was triturated with hexanes to precipitate the product from solution. Filtratration and
Example 310
3-[6-(3-Methoxy-propylamino)-2-propyl-2H-pyrazolo[3,4-d]pryrimidin-4-yl]-5-methyl-
1 3-dihydro-indol-2-one
Using microwave procedure as described for Example 309, the yellow precipitate was
recovered by filtration, and the solid washed with water to provide the product (25 mg,
Compound 312
6,7-Dihydro-5H-cyclopentapyrimidine-2,4-diol
To a 100 mL of flask was added ethyl 2-oxocyclopentanecarboxylate 311 (10 mL, 67.2
mmol), urea (6.07 g, 101 mmol), ethanol (20 mL) and concentrated HC1 (1 mL). After
the mixture was heated to reflux for 2 h, it was cooled to room temp. The ethanol was
decanted and remain white crystalline was heated to reflux in 5% NaOH solution (25
mL) for 30 min. The reaction was cooled to room temp and the precipitate was
collected by filtration. It was washed with water and dried to give 6.77 g (66%) of the
title compound 312._Compound 312: 1H NMR (400 MHz, DMSO-d6) d 11.03 (s, 1H),
10.72 (s, 1H), 2.63 (m, 2H), 2.44 (tn, 2H), 1.98-1.91 (m, 2H); MS (m/e) 153 (M + 1).
Compound 313
2,4-Dichloro 6,7-dihyro-5H-cyclopentapyrimidine
To a 100 mL flask was added compound 312 (3.00 g, 19.7 mmol) and POCl3 (15 mL).
The reaction mixture was heated to reflux for 5 h. After cooled to room temp, the
reaction was concentrated in vacuo. The gummy residue was quenched with ice-water
and the resulted precipitation was collected by filtration. It was washed with water and
dried to give 3.10 g (83%) the title compound 313. Compound 313: 1HNMR(400
MHz, DMSO-d6) d 3.03 (m, 2H), 2.93 (m, 2H), 2.17-2.09 (m, 2H); MS (m/e) 190 (M +
1)-
Example 314
5-Bromo-3-(2-chloro-6,7-dihydro-5H-cyclopentapyrimidin-4-yl)-1,3 -dihydro-indol-2-
one
To a stirring mixture of NaH (480 mg, 12.0 mmol) in THF (20 mL) at 0°C was added 5
bromooxindole (1.00 g, 4.72 mmol) in portion. Additional THF (5mL x 3) was used to
make sure all the oxindole was added into the reaction flask. After stirred for 50 min, a
solution of compound 313 (892 nig, 4.72 mmol) in THF (5 mL x 3) was added. The
reaction was continued stir for 1 h at 0°C and 2.5 h at room temp. A saturated NH4CI
solution (50 mL) was added into the reaction and the mixture was extracted with EtOAc
(50 mL x 3). The combined organic extracts was washed with brine and concentrated.
The residue was triturated with MeOH and (dried to give 1.27 g (74%) the title Example
314. Example 314: 1H NMR (400 MHz, DMSO-d6) d 10.83 (s, 1H), 7.43 (m, 1H),
7.27 (s, 1H), 6.87 (m, 1H), 5.11 (s, 1H), 3.02-2.76 (m, 4H), 2.13-2.03 (m, 2H); MS
(m/e) 365 (M + 1), 366 (M + 2).
Example 315
3-(2-Chloro-6,7-dihydro-5H-cyclopentapyrimidin-4-yl)-5-fluoro-1, 3-dihydro-indol-2-
one
Example 316
5-Chloro-3-(2-chloro-6,7-dihyd ro-5H-cyclopentapyrimidin-4-yl)-l ,3-dihydro-inodol-2-
one
Example 317
3-(2-Chloro-6,7-dihydro-5H-cyclopentapyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
To mixture of 5- cyanooxindole (1.00 g, 6.32 mmol) and NaH (650 mg, 16.3 mmol) was
added THF (20 mL). After the reacton mixture was stirred for 45 min at it, a solution of
313 (1.20 g, 6.32 mmol) in THF (10 mL) was added. The reaction was heated to reflux
for 2 h, and was cooled to room temp. Water (30 mL) was slowly added to the reaction,
and was acidified to pH ~ 3 with concentrated HC1. The resulted precipite was collected
by filtration. It was washed with water, MeOH, and dried to give 1.31 g (67%) of the
desired Example 317. 1H NMR (400 MHz, DMSO-d6) d 11.18 (s, 1H), 7.73 (m, 1H),
7.57 (s. 1H), 7.06 (m, 1H), 5.17 (s, 1H), 3.01 (m, 2H), 2.83 (m, 2H), 2.10 (m, 2H); MS
(m/e)311(M+l).
The following Examples 318-354 in Table 5 were prepared according to procedures
disclosed herein including using methods generally known to one skilled in the art.
Table 5
A mixture of 2-aminonicotinic acid (5.00 g, 36.2 mmol) and urea (10.9 g, 181 mmol) in
a 100 mL flask was heated to 195ºC for 1.5 h. After the reaction was cooled to room
temp, NaOH (1.45g, 36.2 mmol) and water (50 mL) was added. The mixture was
heated to reflux for 1 h and cooled to room temp. The reaction solution was acidified to
pH 4, the resulting precipitate was collected by filtration. It was washed with water and
dried to give 4.49 g (76%) the title Compound 355. '1H NMR (400 MHz, DMSO-^j) d
11.66 (s, 1H), 11.45 (s, 1H), 8.60 |m, 1H), 8.26 (m, 1H), 7.24 (m, 1H).
Compound 356
2,4-Dicnloro-pyrido[2,3-d]pyrimidine
To a 100 mL flask was added Compound 355 (500 mg, 3.06 mmol), N,N-diethylaniline
(1 mL), and POCl3 (10 mL). The reaction mixture was heated to reflux for 5.5 h. After
cooled to room temp, the reaction was concentrated in vacuo. The residue was
quenched with ice-water (50 mL) and was immediately extracted with CHCl3 (50 mL x
3). The combined organic extracts was washed with water (50 mL), dried (MgSO4),
filtered and concentrated to give crude the title Compound 356. The material was used
for next step without purification.
Example 357
5-Bromo-3-(2-chloro-pyrido[2,3-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
To a stirring mixture of NaH (260 mg, 6.50 mmol) in THF (20 mL) at 0°C was added 5-
bromooxindole (551 g, 2.60 mmol) in portion. Additional THF (5mL x 2) was used to
make sure all the oxindole was added into the reaction flask. After stirred for 1 h, a
solution of crude compound 356 in THF (5 mL x 3) was added. The reaction was
continued stir for 1 h at 0°C and 24 h at room temp. A saturated NH4Cl solution (30
mL) was added into the reaction and the resulted red precipitate was collected by
filtration. It was dried to give 361 mg (37%) the title Example 357. 1H NMR (400
MHz, DMSO-d6) d 10.41 (s, 1H), 10.11 (s, 1H), 8.61 (m, 1H), 8.50 (s, 1H), 7.47 (m,
1H), 7.18 (m, 1H), 6.73 (m, 1H); MS (m/e) 376 (M +1).
Example 358
5-Bromo-3-[6-(2-hydroxy-propylamino)-2-methyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
1,3-dihydroindol-2-one
Example 300 (30 mg, 0.079 mmol) and (R)-(-)-1-amino-2-propanol (62 µL, 0.79 mmol)
were heated in 1 mL EtOH at 130°C in microwave for 10 min. Upon cooling, the
product precipitated in the reaction tube. The resulting solid was filtered and pumped
dry to afford 22 mg (67%) of a bright yellow solid, mp 298-301 °C; MS (ES+calculated:
417.27; found: 417.61, 418.73 M+H). HPLC (91%) purity, retention time 2.484
minutes - Method C); 1H NMR (400 MHz, TFA) d 8.75 (s, 1H), 8.4 (s, 1H), 7.55 (br d,
1H), 7.1 (br s, 1H), 4.5 (m, 1H), 4.2 (s, 2H), 3.8 (m, 1H), 3.7 (m, 1H), 1.6 (d, 2H).
Example 359
5-Bromo-3-[6-(2-methylamino-ethylamino)-2-methyl-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one
Example 300 (30 mg, 0.079 mmol) and N,N-dimethylaminoethylamine (69 mg, 0.79
mmol) were heated in 1 mL EtOH at 130°C in microwave for 10 min. Upon cooling,
the product precipitated in the reaction tube. The resulting solid was filtered and
pumped dry to afford 14 mg (41%) of a brown solid, mp 279-289°C; MS
(ES+calculated: 430.31; found: 430.67, 431.82 M+H). HPLC (88%) purity, retention
Example 360
5-Bromo-3-[2-methyl-6-(2-methylamino-ethylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one hydrochloride salt
Example 300 (40 mg, 0.106 mmol) and N-(3-aminoethyl)-N-methyl carbamic acid t-
butyl ester (184 mg, 1.06 mmol) wore heated in 1 mL EtOH at 130°C in microwave for
10 min. Upon cooling, the product precipitated in the reaction tube. The resulting solid
was filtered and pumped dry before stirring in 5 mL of 4N HCl/dioxane for 1 h at RT.
The reaction mixture was pumped dry, triturated in ether and filtered to afford 19 mg
(40%) of a yellow solid, mp 269-272°C; MS (ES+calculated: 416.28; found: 416.58,
417.79 M+H). HPLC (88%) purity, retention time 2.357 minutes - Method C); 1H
NMR (400 MHz, TFA) d 8.74 (s, 1H), 7.89 (s„ 1H), 7.54 (d, J= 8 Hz, 1H), 7.17 (d, J=
8 Hz, 1H), 4.32 (m, 1H), 4.25 (s, 4H), 4.07 (m, 1H), 3.85 (m, 3H), 3.11 (br s, 2H), 3.07
(s, 1H).
Example 361
5-Bromo-3-{2-propyl-6-[((S)-1-pyrrolidin-2-ylmethyl)-amino]-2H-pyrazolo[3,4-
d]pyrimidin-4-yl}-1,3-dihydro-indol-2-one
Example 188 (30 mg, 0.0737 nunol) and (S)-(+)-2-(aminomethyl)pyiTolidine (79 µL,
0.737 mmol) were heated in 1 mL EtOH at 130°C in microwave for 10 min. Upon
cooling, the product precipitated in the reaction tube. The resulting solid was filtered
and pumped dry to afford 15 mg (43%) of a yellow solid, mp 286-293°C; MS
(ES+calculated: 470.38; found: 470.65, 471.81 M+H). HPLC (97%) purity, retention
time 3.66 minutes - Method C); 1H NMR (400 MHz, DMSO-d6) d 9.57 (s, 1H), 9.39 (s,
1H), 8.57 (s, 1H), 7.85 (br s, 4 H), 6.80 (d, .J= 8 Hz, 1H), 6.59 (d, J = 8 Hz, 1H), 4.18
(m, 1H), 4.09 (t, J= 7 Hz, 2H), 186 (m, 1H), 3.74 (m, 1H), 3.07 (d, 7= 12 Hz, 1H),
2.94 (m, 1H), 2.14 (m, 1H), 2.01 (n, 1H), 1.98 (m, 1H), 1.85 (m, 3H), 0.85 (t, J= 8 Hz,
3H).
Example 362
5-Bromo-3-[2-propyl-6-((S)-pyrrolidin-3-ylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
1,3 -dihydro-indol-2-one
Example 188 (40 mg, 0.0983 nunol) and (S)-(-)-3-aminopyrrolidine (87 µL, 0.983
mmol) were heated in 1 mL EtOH at 130°C in microwave for 10 min. Upon cooling,
the product precipitated in the miction tube. The resulting solid was filtered and
pumped dry to afford 44 mg (98%) of a yellow solid, mp 315-320°C; MS
(ES+calculated: 456.35; found: 456.63, 457.79 M+H). HPLC (99%) purity, retention
time 3.40 minutes - Method C); 1H NMR (400 MHz, DMSO-d6) d 9.70 (s, 1H), 9.40 (s,
1H), 8.55 (s, 1H), 6.86 (d, J= 8 Hz, 1H), 6.61 (d, J = 8 Hz, 1H), 5.9 (br s, 2H), 4.11 (t, J
Example 363
5-Bromo-3-[2-propyl-6-((R)-pyrrolidin-3-ylamino)-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-
l,3-dihydro-indol-2-one
Example 188 (45 mg, 0.11 mmol) and (R)-(+)-3-aminopyrrolidine (96 µL, 1.1 mmol)
were heated in 1 mL EtOH at 130°C in microwave for 10 min. Upon cooling, the
product precipitated in the reaction tube. The resulting solid was filtered and pumped
dry to afford 46 mg (92%) of a yellow solid, mp 314-320°C; MS (ES+calculated:
456.35; found: 456.63, 457.78 M+H). HPLC (96%) purity, retention time 3.389
minutes - Method C); 1H NMR (400 MHz, DMSO-d6) d 9.71 (s, 1H), 9.40 (s, 1H), 8.55
(s, 1H), 6.87 (d, J= 8 Hz, 1H), 6.52 (d, J= 8 Hz, 1H), 4.11 (t, 7= 7 Hz, 2H), 3.77 (br s,
2H), 3.70 (br s, 2H), 3.43 (m, 2H) 2.18 (m, 1H), 1.86 (m, 4H), 0.85 (t, J= 7 Hz, 3H).
Example 364
5-Bromo-3-[6-((S)-2,3-dihydroxy-propylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-
4-yl]-1,3-dihydro-indol-2-one
Example 188 (40mg, 0.0983 mmol) and (S)-(-)-3-amino-l,2-propanediol (89 mg, 0.983
mmol) were heated in 1 mL EtOH at 130°C in microwave for 10 min. Upon cooling,
the product precipitated in the reaction tube. The resulting solid was filtered and
pumped dry to afford 28 mg (62%) of a yellow solid, mp 295-297°C; MS
(ES+calculated: 461.32; found: 461.64,462.71 M+H). HPLC (93%) purity, retention
time 3.517 minutes - Method C); 1H NMR (400 MHz, TFA) d 8.74 (s, 1H), 7.90 (s,
Example 365
5-Bromo-3-[6-(2-methylaminoethylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-
yl]-1,3-dihydro-indol-2-one hydrochloride salt
Example 188 (50 mg, 0.123 mmol) and N-(3-aminoethyl)-N-methyl carbamic acid t-
butyl ester (214 mg, 1.23 mmol) were heated in 1 mL EtOH at 130°C in microwave for
10 min. Upon cooling, the product precipitated in the reaction tube. The resulting solid
was filtered and pumped dry before stirring in 5 mL of 4N HCl/dioxane for 1 h at r.t.
The reaction mixture was pumped dry, triturated in ether and filtered to afford 38 mg
(65%) of a yellow solid, mp 269-271°C; MS (ES+calculated: 444.34; found: 444.63,
Example 366
5-Bromo-3 -(2-propyl-6- {[(R)-1-(tetraliydro-furan-2-yl)methyl]-amino} -2H-
pyrazolo[3 J4-d]pytimidin-4-yl)-1,3-dihydro-indoI-2-one
Example 188 (40 mg, 0.0983 mmol) and (R)-(0-tetrahydrofurfurylamine (101 µL, 0.983
mmol) were heated in 1 mL EtOH at 130°C in microwave for 10 min. Upon cooling,
the product precipitated in the reaction tube. The resulting solid was filtered and
pumped dry to afford 26 mg (56%) of a yellow solid, mp 309-312°C; MS
Example 367
5-Bromo-3-(2-propyl-6-{[(S)-1-(tetrahydro-furan-2-yl)methyl]-amino}-2H-
pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Example 188 (40 mg, 0.0983 nimol) and (S)-(+)-tetrahydrofurfurylamine (101 µL,
0.983 mmol) were heated in 1 mL EtOH at 130°C in microwave for 10 min. Upon
cooling, the product precipitated m the reaction tube. The resulting solid was filtered
and pumped dry to afford 32 nig (69%) of a yellow solid, mp 309-312°C; MS
(ES+calculated: 471.36; found: 471.63, 472.78 M+H). HPLC (98%) purity, retention
Example 368
5-Bromo-3-[6-((R)-2,3-dihydroxy-propylamino)-2-propyl-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one
Example 188 (40mg, 0.0983 mmol) and (R)-(+)-3-amino-l,2-propanediol (89 mg, 0.983
mmol) were heated in 1 mL EtOH at 130°C in microwave for 10 min. Upon cooling,
the product precipitated in the reaction tube. The resulting solid was filtered and
Example 369
3-[6-(2-Amino-propylamino)-2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl]-5-bromo-
1,3-dihydro-indol-2-one
(R)-(+)-l,2-diaminopropane dihyirochloride (144 mg, 0.983 mmol) was stirred with
triethylamine in 2 mL EtOH. Once homogeneous Example 188 (40mg, 0.0983 mmol)
was added and the reaction wati heated at 130°C in microwave for 10 min. The
resulting reaction mixture was concentrated onto silica gel and flash columned (10%
MeOH / 1% NH4OH in CH2Cl2). The resulting fractions were concentrated to afford 13
mg (30%) of a yellow solid, mp 247-250°C; MS (ES+calculated: 444.34; found:444.65,
445.76M+H). HPLC (100%) purity, retention time 11.643 minutes - Method B); 1H
NMR (400 MHz, TFA) d 8.74 (s, 1H), 7.90 (s, 1H), 7.56 (d, J= 8 Hz, 1H), 7.17 (d, J=
8 Hz, 1H), 4.44 (m, 2H), 4.23 (br s, 1H), 4.12 (br s, 2H), 2.12 (m, 2H), 1.72 (m, 3H),
1.13(t,J=7Hz,3H).
The following Examples 270-379 in Table 6 were prepared according to procedures
disclosed herein including using methods generally known to one skilled in the art.
Table 6
Example 370
5-Bromo-3-[2-cyclopentyl-6-(2-dimethylamino-ethylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-l ,3-dihydro-indol-2-one
A mixture of 5-Bromo-3-(6-chloro-2-cyclopentyl-2-H-pyrazolo[3,4-d]pyrimidin-4-yl)-
l,3-dihydro-indol-2-one (30 mg, 0.07 mmol) (Example 182), N,N-
dimethylethylenediamine (61mg, 0.7 mmol) and ethanol (2 mL) were heated to 130 °C
in a microwave for 10 minutes. Tht: reaction was concentrated, treated with ethyl ether
and filtered to give 15mg (44%) of Example 370. Example 370: 1HNMR (400MHz,
TFA-d) d 8.8 (s, 1H), 8.0 (s, 1H), 7 6 (d, 1H), 7.3 (d, 1H), 5.1 (m, 1H), 4.4 (m, 2H), 4.0
(m, 3H), 3.3-3.4 (m, 6H), 2.0-2.7 (m, 8H); MS (m/e) 484 (M + 1); HPLC (87%) purity,
retention time 3.052 minutes - Method C; mp 190-192°C.
Example 371
A mixture of 5-Bromo-3-(6-chloro-2-cyclopentyl-2-H-pyrazolo[3,4-d]pyrimidin-4-yl)-
l,3-dihydro-indol-2-one (50 mg, 0.115 ramol) (Example 182), N'-BOC-N'-
methylethylenediamine (200 nig, 1.15 mmol) and ethanol (2 mL) were heated to 130 °C
in a microwave for 10 minutes. The reaction was concentrated, treated with ethyl ether
and filtered to give 36mg (55%) of the product. The product was dissolved in 4N HC1 in
dioxane (3mL) and stirred at rt for lhr. The reaction was concentrated, treated with
acetone and filtered. The solid was washed with acetone, ethyl ether and pumped dry to
give 9mg (28%) of Example 378. Example 378: 1HNMR (400MHz, TFA-d) d 8.9 (s,
1H), 8.1 (s, 1H), 7.7 (d, 1H), 7.4 (d, 1H), 5.1 (m, 1H), 4.3 (m, 2H), 4.0 (m, 4H), 3.1 (m,
3H), 2.1-2.8 (m, 8H); MS (m/e) 470 (M + 1); HPLC (99%) purity, retention time 3.040
minutes - Method C; mp 201-204°C.
Example 379
5-Bromo-3-[2-cyclopentyl-6-(3-memylamino-propylamino)-2H-pyrazolo[3,4-
d]pyrimidin-4-yl]-1,3-dihydro-indol-2-one hydrochloride salt
Example 379 was synthesized ir a similar manner to Example 378 using the approriate
starting materials. Example 379: 1HNMR (400MHz, TFA-d) d 8.9 (s, 1H), 8.1 (s, 1H),
7.7 (d, 1H), 7.4 (d, 1H), 5.1 (m, 1H), 4.3 (in, 2H), 3.5 (m, 8H), 2.1-2.8 (m, 8H); MS
(m/e) 484 (M + 1); HPLC (99%) purity, retention time 3.003 minutes - Method C; mp
208-211°C.
Scheme 9 discloses a general procedure for the preparation of compounds of the
invention wherein R6 is an alklyl group.
Scheme 9
Example 380
5-Bromo-3-(2-isopropyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Example 380 was synthesized in using the general procedure of Scheme 9, wherein R2 =
Br, and the approriate starting materials. Example 380: 1HNMR (400MHz, TFA-d)
d 9.1 (s, 1H), 8.8 (s, 1H), 7.9 (s, 1H), 7.6 (d, 1H), 7.2 (d, 1H), 5.0 (m, 1H), 1.8 (m, 6H);
MS (m/e) 373 (M + 1); HPLC (99%) purity retention time 3.298 minutes - Method C;
mp >300°C.
Example 381
5-Chloro-3-(2-isopropyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one
Example 381 was synthesized in using the general procedure of Scheme 9, wherein R2 =
Cl, and the approriate starting materials. Example 381: !HNMR (400MHz, TFA-d)
d 9.1 (s, 1H), 8.9 (s,lH), 7.8 (s, l H), 7.4 (d, 1H), 7.3 (d, 1H), 5.0 (m, 1H), 1.8 (m, 6H);
MS (m/e) 328 (M + 1); HPLC (99%) purity retention time 3.189 minutes - Method C;
mp >300°C.
Example 382
3-(2-Isopropyl-2H-pyrazolo[3,4-d]pyri;midin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile
Example 382 was synthesized in using the general procedure of Scheme 9, wherein R2 =
CN, and the approriate starting materials. Example 382: lHNMR (400MHz, TFA-d)
d 10.3 (s, 1H), 9.0 (s, 1H), 8.3 (s, 1H), 7.9 (d, 1H), 7.5 (d, 1H), 5.0 (m, 1H), 1.8 (m, 6H);
MS (m/e) 319 (M + 1); HPLC (99%) purityretention time 3.189 minutes - Method C;
mp >300°C.
Example 383
5-Bromo-3-fluoro-3-(2-propyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-l,3-dihydroindol-2-
one
To a solution of Example 97 (85 mg, 0.23 mmol) in a mixture of THF7 dioxane (1:1.
12.8 mL) was added a 1M solution of sodium bis(trimethylsilyl)amide (0.23 ml) at -40
oC followed by addition of l-fluoro-2,4,6-trimethylpyridinium triflate (67 mg, 0.23
mmol). The reaction was allowed to warm to room temperature where it stirred
overnight. The reaction was heated to ~50 °C for 4h then quenched with ammonium
chloride. The reaction was concentrated in vacuo and purified prep HPLC on a Rainin
Dynamax system with a Higgins Analytical Clipeus 10 um CI8 column (250 x 20 mm).
Example 383: 1HNMR (400MHz, DMSO-d6) d 0.91 (s, 3H), 2.02 (m, 2H), 4.54 (m,
2H), 6.98 (d, 1H), 7.53 (s, 1H), 7.61 (d, 1H), 8.85 (s, 1H), 9.03 (s, 1H), 11.2 (s, 1H); MS
(m/e) 391 (M + 1); HPLC (91%) purity retention time 3.87 minutes - Method F; mp
184-186 °C.
HPLC Methods:
Wavelengths monitored included 254,290 and /or 215 nm
Method A: Flow rate: 1.6 mL/min, gradient over 15 minutes from 10 to 50%
[acetonitrile (0.1% TFA added):water (0.1% TFA added)] ramping with a gradient from
50% to 100% acetonitrile:water from 15 to 20 minutes, column: 5 micron Zorbax RX-
C8 (4.6X150 mm).
Method B: Flow rate 1.6 mL/min, gradient over 20 minutes from 10 to 100%
[acetonitrile (0.1% TFA added):water (0.1% TFA added)], column: 5 micron Zorbax
RX-C8 (4.6X150 mm).
Method C: Flow rate 2.4 mL/min, gradient over 8 minutes at 30°C from 10 to 100%
[acetonitrile (0.1% TFA added):water (0.1% TFA added)], column: 3.5 micron Zorbax
SB-C18 (4.6X75 mm).
Method D: Flow rate 1.6 mL/min, 100% water (0.1% TFA added) for 1 minute,
gradient over 15 minutes from 0 to 100% [acetonitrile (0.1% TFA added):water] 100%
acetonitrile for 4 minutes, column 5 micron Zorbax RX-C8 (4.6 X 150 mm).
Method E: Flow rate 1.6 mIV min, 10-100% Acetonitrile/ water (both with 0.1% TFA)
over 7 minutes.
Method F: Flow rate 1.6 mL/ min, 10-100% Acetonitrile/ water (both with 0.1% TFA)
over 8 minutes.
It is noted that NMRs of some oxindole containing products (particularly in the N-2
series) are very complicated due to possible restricted rotational isomerism as well as
possible tautomeric forms - signal positions only and no integrations are given for the
N-2 series due to inability to assign protons.
Utility
The present invention relates to novel substituted heterobicyclic pyrimidine
compounds, in particular substituted pyrazolopyrimidine oxindoles, that act as inhibitors
of glycogen synthase kinase 3 and/or cyclin dependant kinase 5, and their use in the
treatment of chronic neurodegenerative diseases, neurotraumatic diseases, depression
and/or diabetes. Compounds of the present invention are well suited as inhibitors of
GSK-3ß activity and/or CDK5 activity. Representative compounds of the invention
have exhibited good in vitro potency against GSK-3ß kinase and/or CDK5 kinase. Table
7 below provides data related to several example compounds of the invention with
respect to, for example, ability to inhibit GSK-3ß activity and/or CDK5 activity.
Accordingly, the compounds of the present invention are expected to be useful in the
prention and/or treatment of conditions mediated by GSK-30 activity and/or CDK5
activity.
Cloning, Expression and Purification of CDK5/GST-p25
Two recombinant baculoviral constructs were created, one encoding for human
CDK5 and the other encoding for human p25 with an arnino-terminal glutathione-S-
transferase (GST) tag. PCR amplification of full-length CDK5 was performed utilizing
human brain cDNA as a template and Pfu Turbo polymerase (Strategene). The PCR
product from this reaction was subcloned into the baculoviral expression vector
pFASTBACl (Gibco/BRL). The final construct, encoding for full-length human CDK5
(base pairs 25-903 of GenBank Accession #NM_004935), is 292 amino acids long with
a predicted MW of 33.3 kDa.
For p25, the active truncated form of p35, amino acids 108-307 were PCR-
amplified from human fetal brain cDNA (Clontech QUICK-Clone cDNA) using the
Advantage 2 PCR system (Clontech). The PCR product was subcloned into the
baculoviral transfer vector pFBGSTP (an engineered derivative of the baculoviral
transfer vector pFASTBACl). The final baculoviral construct encodes for base pairs
419-1021 of GenBank Accession ;MNM_003885, with an amino-terminal GST tag. The
expressed GST-p25 is 444 amino acids long with a predicted MW of 50.3 kDa.
The CDK5/GST-p25 complex was generated by coexpression. Sf21 cells were
cultured in TNM-FHS media at a density of 1.5 x 106 cells/mL and infected with each
recombinant virus at MOI values of 5 (for CDK5) and 10 (for GST-p25). The cells
were harvested 40 h after infection For purification, the 100,000 x g supernatant
solution was used. Expression was confirmed by running samples on SDS-PAGE,
followed by immunoblot analysis utilizing antibodies against CDK5 (anti-CDK5 (268-
283); Calbiochem #219449) and p35 (Santa Cruz #sc820). The CDK5/GST-p25
complex was purified by glutathione affinity chromatography.
Inhibition of CDK5/GST-p25 Kinase Activity
Compounds were tested for their ability to inhibit the kinase activity of .
recombinant baculoviral CDK5/GST-p25 using an enzyme-linked immunosorbent assay
(ELISA) with time-resolved fluorescence (TRF) readout. Briefly, each 384-well
FluoroNunc Maxisorp plate (Cat # 460372) plate was coated with 50 µl/well of 50
ug/ml substrate solution (recombinant GST-Rb(773-928)) in Tris-buffered saline (TBS).
The CDK5/GST-p25 assay mixture (total volume = 50 µl/well) consisting of 20 mM
HEPES (pH 7.2), 10 uM ATP, 10 mM MgCl2,5 mM EGTA, 25 mM p-
glycerophosphate, 0.05% BSA, 2.5% DMSO, and various concentrations of test
compound were then added to the assay plate. Enzyme (2 ng/ml CDK5/GST-p25) was
added and the reaction was allowed to proceed at 37°C for 20 minutes. Detection of the
phosphorylated product was performed by adding 50 µl/well of phospho-Rb (Ser-780)
antibody (Cell Signaling # 9307) diluted 1:10,000 in antibody dilution buffer (0.1%
BSA in TBST). After 1-hour incubation at 37°C, 50 µl/well of Eu-Nl labelled anti-
rabbit antibody (Wallac # AD01C5; 1:50,000 in antibody dilution buffer) was added.
Incubation at 37°C then proceeded for 1 hour, followed by addition of 50 µl
enhancement solution (Wallac #1244-105). The plate was gently agitated and after a few
minutes, the fluorescence of the resulting solution was measured using a Multilabel
Reader (Victor2 Model # 1420-018 or Envision Model # 2100). Inhibition data were
analyzed using ActivityBase and IC50 curves were generated using XLFit 3.0.5.
Cloning, Expression and Purification of His6-GSK-3ß
Full-length GSK-30 was amplified from a sequence-verified I.M.A.G.E. EST
acquired from Research Genetics (Invitrogen, Clone ID# CS0DB003YJ02). The final
sequence-verified cDNA contained the coding region for a NH2-terminal tag, which
encoded for 6 histidines and then eight vector-encoded amino acids prior to the start of
GSK3ß, which contained bp # 43-342 of Genbank Accession # NM_002093, encoding
amino acids #2-419. The predicted molecular weight of the tagged, 435 amino acid,
full-length protein is 48.5 kDa. The major structural element of this protein is the kinase
domain, which is from amino acids #56-340. Recombinant baculoviral DNA was
prepared by transposition in E. coli (BAC-TO-BAC system: Invitrogen) and the virus
generated and amplified in Sf21 insect cells. A suspension culture of Sf21 cells was
infected at an MOI of 0.7 and cell density of 1.5 x 106 cells/mL in Excell 420 serum-free
media (JRH BioScience) and harvested 65 h after infection. The 100,000 x g
supernatant solution was used for purification. Expression was confirmed by running
samples on SDS-PAGE, followed by immunoblot analysis utilizing both a Penta-HIS
antibody (Qiagen #34660) and a GSK-3a/GSK-3ß antibody (Calbiochem #368662, data
not shown). The His6-tagged protei a was purified to in one step by Ni-NTA affinity
chromatography.
Inhibition of His6-GSK-3ß Kinase Activity
Inhibitory effects of compounds on baculoviral GSK-3ß kinase activity were
evaluated using an ELISA-based format in a 384-well FluoroNunc Maxisorp plate (Cat
# 460372) with a time-resolved fluorescence readout. Briefly, each plate was coated with 50 µl/well of 20 µg/ml substrate solution (recombinant GST-Rb) in Tris-buffered
saline (TBS). The GSK-3ß assay mixture (total volume = 50 µl/well) consisting of 50
mM HEPES (pH 7.2), 20 µM ATP, 10 mM MgCl2,5 mM EGTA, 25 mM ß-
glycerophosphate, 0.05% BSA, 2.5% DMSO, and various concentrations of test
compound were then added to the: assay plate. Enzyme (200 ng/ml His6-GSK-3ß) was
then added and the reaction was allowed to proceed at 37°C for 30 minutes. Detection of
the phosphorylated product was performed by adding 50 µl/well of phospho-Rb (Ser-
780) antibody (Cell Signaling # 9307) diluted 1:10,000 in antibody dilution buffer
(0.1% BSA in TBST). After 1-hour incubation at 37°C, 50 µl/well of Eu-Nl labelled
anti-rabbit antibody (Wallac # AD0105; 1:50,000 in antibody dilution buffer) was
added. Incubation at 37°C then proceeded for 1 hour, followed by addition of 50 µl
enhancement solution (Wallac #1244-105). The plate was gently agitated and after a few
minutes, the fluorescence of the resulting solution was measured using a Multilabel
Reader (Victor2 Model # 1420-018 or Envision Model 2100). Inhibition data were
analyzed using ActivityBase and IC50 curves were generated using XLFit 3.0.5.
Compound Activity
Using the assays disclosed herein the following Table 3 demonstrates the utility
of compounds of the invention for tau kinase inhibition. Compounds of the present
invention are considered active if their IC50 values are less than 50 uM. In the following
Table, for the inhibition of CDK5, compounds of the present invention with a "+" are
less than 10000 nM; compounds of the present invention with a "++" are less than 3000
nM; and compounds of the present invention with a "+++" are less than 300 nM in IC50
for CDK5 inhibition. In the following Table, for the inhibition of GSK3ß, compounds of
the present invention with a "+" are less than 10000 nM; compounds of the present
invention with a "++" are less than 3000 nM; and compounds of the present invention
with a "+++" are less than 300 nM in IC50 for GSK3ß inhibition. Where ">+" occurs
activity was greater than the limits of the assay. Where no IC50 value is represented, data
has yet to be determined.
Accordingly, these results demonstrate that compounds of the present invention exhibit
inhibitory activity against GSK3ß kinase and/or CDK5 kinase.
References:
1. Bacon, Edward R.; Singh, Baldev; L esher, George Y. 6-(heterocyclyl)pvrazolo[3,4-d]pvrirnidin-4-
one phosphodiesterase inhibitors. (1994), US 5294612 A.
2. Herling, Andreas; Maguire, Martin P.; Spada, Alfred P.; Myers, Michael R.; Chot-Sledeski, Yong
Mi; Pauls, Heinz W.; Ewing, William R. Adenosine analogues for the treatment of insulin resistance
syndrome and diabetes. (2001), Ep Appl. 1 258 247 Al
3. Chu, I. Lynch, B.M. Synthesis and Biological evaluation of Xanthine Oxidase Inhibitors. Pyrazolo[3,4-
d]pyrimidines andPyrazolo[3,4-b]pyndines. J. Med Chem. 1975, 18, 161-165.
Pharmaceutical Formulations and Dosage Forms
When employed as pharmaceuticals, the compounds of Formula (I) can be
administered in the form of pharmaceutical compositions. These compositions can be
administered by a variety of routes including oral, rectal, transdermal, subcutaneous,
intravenous, intramuscular, and intranasal, and can be prepared in a manner well known
in the pharmaceutical art.
This invention also include.1: pharmaceutical compositions which contain, as the
active ingredient, one or more of the compounds of Formula (I) above in combination
with one or more pharmaceutically acceptable carriers. In making the compositions of
the invention, the active ingredient is typically mixed with an excipient, diluted by an
excipient or enclosed within such a carrier in the form of, for example, a capsule, sachet,
paper, or other container. When the excipient serves as a diluent, it can be a solid, semi-
solid, or liquid material, which acts as a vehicle, carrier or medium for the active
ingredient. Thus, the compositions can be in the form of tablets, pills, powders,
lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols
(as a solid or in a liquid medium), ointments containing, for example, up to. 10% by
weight of the active compound, soft and hard gelatin capsules, suppositories, sterile
injectable solutions, and sterile packaged powders.
In preparing a formulaaon, the active compound can be milled to provide the
appropriate particle size prior to combining with the other ingredients. If the active
compound is substantially insoluble, it can be milled to a particle size of less than 200
mesh. If the active compound is substantially water soluble, the particle size can be
adjusted by milling to provide a substantially uniform distribution in the formulation,
e.g. about 40 mesh.
Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol,
mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin,
calcium silicate, microcrystallme cellulose, polyvinylpyrrolidone, cellulose, water,
syrup, and methyl cellulose. The formulations can additionally include: lubricating
agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying
and suspending agents; preserving agents such as methyl- and propylhydroxy-benzoates;
sweetening agents; and flavoring agents. The compositions of the invention can be
formulated so as to provide quick, sustained or delayed release of the active ingredient
after administration to the patient by employing procedures known in the art.
The compositions can be formulated in a unit dosage form, each dosage
containing from about 5 to about 100 mg, more usually about 10 to about 30 mg, of the
active ingredient. The term "unit dosage forms" refers to physically discrete units
suitable as unitary dosages for human subjects and other mammals, each unit containing
a predetermined quantity of active material calculated to produce the desired therapeutic
effect, in association with a suitable pharmaceutical excipient.
The active compound can be effective over a wide dosage range and is generally
administered in a pharmaceutical ly effective amount. It will be understood, however,
that the amount of the compound actually administered will usually be determined by a
physician, according to the relevant circumstances, including the condition to be treated,
the chosen route of administration, the actual compound administered, the age, weight,
and response of the individual patient, the severity of the patient's symptoms, and the
like.
For preparing solid compositions such as tablets, the principal active ingredient
is mixed with a pharmaceutical excipient to form a solid preformulation composition
containing a homogeneous mixture of a compound of the present invention. When
referring to these preformulation compositions as homogeneous, the active ingredient is
typically dispersed evenly throughout the composition so that the composition can be
readily subdivided into equally effective unit dosage forms such as tablets, pills and
capsules. This solid preformulation is then subdivided into unit dosage forms of the type
described above containing fiom, for example, 0.1 to about 500 mg of the active
ingredient of the present invention.
The tablets or pills of the present invention can be coated or otherwise
compounded to provide a dosage form affording the advantage of prolonged action. For
example, the tablet or pill can comprise an inner dosage and an outer dosage component,
the latter being in the form of an envelope over the former. The two components can be
separated by an enteric layer which serves to resist disintegration in the stomach and
permit the inner component to pass intact into the duodenum or to be delayed in release.
A variety of materials can be used for such enteric layers or coatings, such materials
including a number of polymeric acids and mixtures of polymeric acids with such
materials as shellac, cetyl alcohol, and cellulose acetate.
The liquid forms in which me compounds and compositions of the present
invention can be incorporated foi administration orally or by injection include aqueous
solutions, suitably flavored syrup:!, aqueous or oil suspensions, and flavored emulsions
with edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as
elixirs and similar pharmaceutical vehicles.
Compositions for inhalation or insufflation include solutions and suspensions in
pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and
powders. The liquid or solid compositions may contain suitable pharmaceutically
acceptable excipients as described supra. In some embodiments, the compositions are
administered by the oral or nasal respiratory route for local or systemic effect.
Compositions in can be nebulized by use of inert gases. Nebulized solutions may be
breathed directly from the nebulizing device or the nebulizing device can be attached to
a face masks tent, or intermittent positive pressure breathing machine. Solution,
suspension, or powder composition; can be administered orally or nasally from devices
which deliver the formulation in an appropriate manner.
The amount of compound or composition administered to a patient will vary
depending upon what is being administered, the purpose of the administration, such as
prophylaxis or therapy, the state of the patient, the manner of administration, and the
like. In therapeutic applications, compositions can be administered to a patient already
suffering from a disease in an amount sufficient to cure or at least partially arrest the
symptoms of the disease and its complications. An amount adequate to accomplish this
is referred to as "therapeutically effective amount." Effective doses will depend on the
disease condition being treated as well as by the judgement of the attending clinician
depending upon factors such as the severity of the disease, the age, weight and general
condition of the patient, and the like.
The compositions administered to a patient can be in the form of pharmaceutical
compositions described above;. These compositions can be sterilized by conventional
sterilization techniques, or may be sterile filtered. Aqueous solutions can be packaged
for use as is, or lyophilized, the lyophilized preparation being combined with a sterile
aqueous carrier prior to administration. The pH of the compound preparations typically
will be between 3 and 11, mors preferably from 5 to 9 and most preferably from 7 to 8.
It will be understood that use of certain of the foregoing excipients, carriers, or
stabilizers will result in the formation of pharmaceutical salts.
The therapeutic dosage of the compounds of the present invention can vary
according to, for example, the particular use for which the treatment is made, the
manner of administration of the compound, the health and condition of the patient, and
the judgment of the prescribing physician. The proportion or concentration of a
compound of the invention in a pharmaceutical composition can vary depending upon a
number of factors including dosage, chemical characteristics (e.g., hydrophobicity), and
the route of administration. For example, the compounds of the invention can be
provided in an aqueous physiological buffer solution containing about 0.1 to about 10%
w/v of the compound for parenteral adminstration. Some typical dose ranges are from
about 1 ug/kg to about 1 g/kg of body weight per day. In some embodiments, the dose
range is from about 0.01 mg/kg >:o about 100 mg/kg of body weight per day. The dosage
is likely to depend on such variables as the type and extent of progression of the disease
or disorder, the overall health status of Ihe particular patient, the relative biological
efficacy of the compound selected, formulation of the excipient, and its route of
administration. Effective doses can be extrapolated from dose-response curves derived
from in vitro or animal model test systems.
The present invention also includes pharmaceutical kits useful, for example, in
the treatment or prevention of inflammatory diseases, which comprise one or more
containers containing a pharmaceutical composition comprising a therapeutically
effective amount of a compound of Formula (I). Such kits can further include, if desired,
one or more of various conventional pharmaceutical kit components, such as, for
example, containers with one or more pharmaceutically acceptable carriers, additional
containers, etc., as will be readily apparent to those skilled in the art. Instructions, either
as inserts or as labels, indicating quantities of the components to be administered,
guidelines for administration, and'or guidelines for mixing the components, can also be
included in the kit.
Various modifications of the invention, in addition to those described herein,
will be apparent to those skilled in the art from the foregoing description. Such
modifications are also intended to fall within the scope of the appended claims. Each
reference cited in the present application, including patents, published patent
applications, and journal articles, is incorporated herein by reference in its entirety.
As those skilled in the art will appreciate, numerous changes and modifications
maybe made to the embodiments of the invention without departing from the spirit of
the invention. It is intended that all such variations fall within the scope of the
invention.
AMENDED CLAIMS
received by the International Bureau on 04 August 2008 (04.08.2008)
1. A compound of Formula (T):
and stereoisomeric forms, mixtures of stereoisomeric forms, tautomeric forms, prodrugs,
or pharmaceutically acceptable salt forms thereof, wherein:
W is CH or N;
ring A is
R1, R2, R3, and R4 at each occurrence are independently selected from
H, halo, -OR11, -NO2, -CN, -CF3, -CHF2, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4
haloalkyl, -NR13R14, -NHOR13a, -C(=O)R15, -C(=O)OR15, -OC(O)R15, -
C(=O)NR13R14, -NR13aC(=O)R15, -NR13aCO2R15, -OC(=O)NR13R14, -
NR13aC(=S)R15, -SR15, -S(=O)R15, -S(=O)2R15, -S(=O)2NR13R14, and C1-C4
alkyl substituted with 0-1 R19;
R5 is H, C1-C6 alkyl or a prodrug of an amino group;
R6 is selected from H;
C1-C6 alkyl substituted by 0-2 R22;
C2-C6 alkenyl substituted by 0-2 R22;
C2-C6 alkynyl substituted by 0-2 R22; and
C3-C7 cycloalkyl substitu ted by 0-3 R22;
R7 is H,-NO2, halo, C1-C4 alkyl or-NR23R24;
X is selected from H, -NR9R10, halo, OR12, -NO2, -CN, -GF3, -CHF2, C,-C4 alkyl,
C2-C4 alkenyl, C2-C4 alkynyl, C1-C4 haloalkyl, -CH2NR9R10, -CH2OR12,
-NHOR16, -C(=O)R18, -C(=O)OR18, -OC(=O)R18, -C(=O)NR9R10, -
NRI6C(=O)R18,
-NR16CO2R18, -OC(=O)NR9R10, -NR16C(=S)R18, -SR18, -S(=O)R18, -S(=O)2R18,
-S(=O)2NR9R10, and -NR16S(=O)2R18;
R9 and R10 at each occurrence are each independently selected from H, -NH2;
C1-C6 alkyl substituted by 0-1 R19;
C2-C6 alkenyl substituted by 0-1 R19:;
C2-C6 alkynyl substituted by 0-1 R19;
C6-C10 aryl substituted by 0-5 R19;
C3-C7 carbocyclyl substituted by 0-5 R19; and
5 to 14 membered heterocyclyl group substituted by 0-5 R19, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaiyl group substituted by 0-5 R19, wherein said
heteroaryl group compiises one, two, or three heteroatoms selected from N,
O, and S;
alternatively, R9 and R10, together with the nitrogen to which they are attached, form a
3-7 membered heterocyclic ring, wherein said 3-7 membered heterocyclic ring
contains a nitrogen atom and optionally a second atom selected from N, O, S,
S(=O), and S(=O)2, wherein said 3-7 membered heterocyclic ring is substituted
with 0-1 R17;
R11 at each occurrence is indepmdently selected from H, C1-C4 alkyl, and C1-C4
haloalkyl;
R12 at each occurrence is independently selected from H, C1-C4 haloalkyl and C1-C4
alkyl substituted with 0-1 R19;
R13 and R14, at each occurrence, are independently selected from H, C1-C4 alkyl
substituted with 0-3 R30; and C6-C10 aryl substituted with 0-5 R30;
R13 a at each occurrence is independently selected from H, C1-C4 alkyl, and C6-C10 aryl;
R15 at each occurrence is independently selected from H,
C1-C6 alkyl substituted by 0-1 R30;
C2-C6 alkenyl substituted by 0-1 R30;
C2-C6 alkynyl substituted by 0-1 R30;
C6-C10 aryl substituted by 0-5 R30;
C3-C7 carbocyclyl substituted by 0-5 R30; and
5 to 14 membered heteroc yclyl group substituted by 0-5 R30, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N,O, andS;
5 to 14 membered heteroaryl group substituted by 0-5 R30, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O, and S;
R16 at each occurrence is independently selected from H and C1-C4 alkyl;
R17 is H, -NR23R24, halo, -NO2, -CN, -CF3, C1-C4 haloalkyl, -NHOH, OR25, C(=O)R25,
C(=O)OR25, OC(=O)R25, C(=O)NR23R24, NR23aC(=O)R25, NR^COjR25,
OC(=O)NR23R24, NR23aC(=S)R25, SR25, S(=O)R25, S(=O)2R25; S(=O)2NR23R24,
-NR23aS(=O)2R25, or C1-C4 alkyl substituted by 0-1 R19;
R18 at each occurrence is independently selected from H;
C1-C6 alkyl substituted by 0-1 R30;
C2-C6 alkenyl substituted by 0-1 R30;
C2-C6 alkynyl substituted by 0-1 R30;
C6-C10 aryl substituted by 0-5 R30;
C3-C7 carbocyclyl substituted by 0-5 R30; and
5 to 14 membered heterocyclyl group substituted by 0-5 R30, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R30, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O,and S;
R19 at each occurrence is independently selected from H, -NR23R24, halo, -NO2, -CN, -
CF3, C1-C4haloalkyl, -NHOH, OR25, C(=O)R25, C(=O)OR25, OC(=O)R25,
C(=O)NR23R24, NR23aC(=O)R25, NR23aCO2R25, OC(=O)NR23R24,
NR23aC(=S)R25, SR25, S(=O)R25, S(=O)2R25; S(=O)2NR23R24, -NR23aS(=O)2R25,
C1-C4 alkyl substituted by 0-1 R30;
C2-C4 alkenyl substituted by 0-1 R30;
C2-C4 alkynyl substituted by 0-1 R30;
C6-C10 aryl substituted by 0- 5 R30;
C3-C7 carbocyclyl substituted by 0-5 R30; and
5 to 14 membered heterocyclyl group substituted by 0-5 R30, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O,and S;
5 to 14 membered heteroaryl group substituted by 0-5 R30, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O, and S;
R22 is H, -NR23R24, -N3, halo, -NO2, -CN, -CF3, C1-C4 alkyl, C2-C4 alkenyl, C2-C4
alkynyl, C1-C4 haloalkyl, C3-C7 carbocyclyl, phenyl, -NHOH, OR25, -CH2OR25,
C(=O)R25, C(=O)OR25,OC(=O)R25, C(O)NR23R24, NR23aC(=O)R25,
NR23aCO2R25, OC(=O)NR23R24,NR23aC(=S)R25, SR25, S(=O)R25, S(=O)2R25;
S(=O)2NR23R24, or -NR23aS(= O)2R25;
R23 and R24 at each occurrence are each independently selected from H or C1-C6 alkyl;
alternatively, R23 and R24, together with the nitrogen to which they are attached, form a
3-7 membered heterocyclic ring, wherein said 3-7 membered heterocyclic ring
contains a nitrogen atom and optionally a second atom selected from N, O and S;
wherein said 3-7 membered heterocyclic ring is substituted with 0-1 C1-C4 alkyl;
R23a at each occurrence is each independently selected from H or C1-C4 alkyl;
R at each occurrence is each independently selected from H or C1-C6 alkyl; and
R30 is H, F, Cl, Br, -CF3, C1-C6 alkyl, and C1-C6 alkoxy;
provided when ring A is

then X is -NR9R10.
2. The compound of Claim 1 wherein ring A is
3. The compound of Claim 1 wherein ring A is
4. The compound of Claim 1 wherein ring A is
5. The compound of Claim 1 wherein R1, R2, R3, and R4 at each occurrence are
independently selected from H, halo, -OR11, -NO2, -CN, and -CF3.
6. The compound of Claim 1 wherein R1, R3, and R4 are each H and R2 is selected
from H, F, Cl, Br, -OCH3, -NO2, -CN, and -CF3.
7. The compound of Claim 1 wherein X is H, -NR9R10, halo, C1-C4 alkyl, or OR12.
8. The compound of Claim 1 wherein X is -NR9R10.
9. The compound of Claim 1 of Formula (II):
and stereoisomeric forms, mixtures of stereoisomeric forms, tautomeric forms, prodrugs,
or pharmaceutically acceptable salt forms thereof, wherein:
R1, R2, R3, and R4 at each occurrence are independently selected from
H, halo, -OR11, -NO2, -CN, -CF3, -CHF2, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4
haloalkyl, and C1-C4 alkyl;
R5 is H, C1-C6 alkyl or a prodrug of an amino group;
R6 is selected from H;
C1-C6 alkyl substituted by 0-2 R22;
C2-C6 alkenyl substituted by 0-2 R22;
C2-C6 alkynyl substituted by 0-2 R22; and
C3-C7 cycloalkyl substituted by 0-2 R22;
R7 is H, -NO2, halo, C1-C4 alkyl or -NR23R24;
X is H, -NRV, halo, OR12, C1-C4 alkyl, or C2-C4 alkenyl;
R9 and R10 at each occurrence are each independently selected from H, -NH2;
C1-C6 alkyl substituted by 0-1 R19;
C2-C6 alkenyl substituted by 0-1 R19;
C2-C6 alkynyl substituted by 0-1 R19;
C6-C10 aryl substituted by 0-5 R19;
C3-C7 carbocyclyl substituted by 0-5 R19; and
5 to 14 membered heterocyclyl group substituted by 0-5 R19, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R19, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O, and S;
alternatively, R9 and R10, together with the nitrogen to which they are attached, form a
3-7 membered heterocyclic ring, wherein said 3-7 membered heterocyclic ring
contains a nitrogen atom and optionally a second atom selected from N, O, S,
S(=O), and S(=O)2, wherein said 3-7 membered heterocyclic ring is substituted
with 0-1 R17;
R11 at each occurrence is independently selected from H, C1-C4 alkyl, and C1-C4
haloalkyl;
R12 at each occurrence is independently selected from H, C1-C4 haloalkyl and C1-C4
alkyl substituted with 0-1 R19;
R17 is H or C1-C4 alkyl substituted by 0-1 R19;
R19 at each occurrence is independently selected from H, -NR23R24, halo, -NO2, -CN, -
CF3, C1-C4 haloalkyl, -NHOH, OR25, C(=O)R25, C(=O)OR25, OC(=O)R25,
C(=O)NR23R24, NR23aC(=O)R25, NR23aCO2R25, OC(=O)NR23R24,
NR23aC(=S)R25, SR25, S(=O)R25, S(=O)2R25; S(=O)2NR23R24, -NR23aS(=O)2R25,
C1-C4 alkyl substituted by 0-1 R30;
C2-C4 alkenyl substituted by 0-1 R30;
C2-C4 alkynyl substituted by 0-1 R30;
C6-C10 aryl substituted by 0-5 R30;
C3-C7 carbocyclyl substituted by 0-5 R30; and
5 to 14 membered heterocyclyl group substituted by 0-5 R30, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R30, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O, and S;
R22 is H, -NR23R24, -N3, halo, -NO2, -CN, -CF3, C1-C4 alkyl, C2-C4 alkenyl, C2-C4
alkynyl, C1-C4 haloalkyl, C3-C7 carbocyclyl, phenyl, -NHOH, OR25, -CH2OR25,
C(=O)R25, C(=O)OR25, OC(=O)R25, C(=O)NR23R24, NR23aC(=O)R25,
NR23aCO2R25, OC(=O)NR23R24, NR23aC(=S)R25, SR25, S(=O)R25, S(=O)2R25;
S(=O)2NR23R24, or-NR23aS(=O)2R25;
R23 and R24 at each occurrence are each independently selected from H or C1-C6 alkyl;
alternatively, R23 and R24, togethtsr with the nitrogen to which they are attached, form a
3-7 membered heterocyclic ring, wherein said 3-7 membered heterocyclic ring
contains a nitrogen atom tmd optionally a second atom selected from N, O and S,
wherein said 3-7 membered heterocyclic ring is substituted with 0-1 C1-C4 alkyl;
R23a at each occurrence is each independently selected from H or C1-C4 alkyl;
R25 at each occurrence is each independently selected from H or C1-C6 alkyl; and
R30 is H, F, Cl, Br, -CF3, C1-C6 alkyl, and C1-C6 alkoxy.
10. The compound of Claim 9 wherein ring A is

11. The compound of Claim 9 wherein ring A is

12. The compound of Claim 9 wherein R1, R2, R3, and R4 at each occurrence are
independently selected from H, F, Cl, Br, -OCH3, -NO2, -CN, and -CF3.
13. The compound of Claim 9 wherein R1, R3, and R4 are each H and R2 is selected
from H, F, Cl, Br, -OCH3, -NO2, -CN, and -CF3.
14. The compound of Claim 9 wherein X is -NR9R10.
15. The compound of Claim 9 wherein R6 is cyclopentyl.
16. The compound of Claim 1 of Formula (III):
and stereoisomers forms, mixtures of stereoisomers forms, tautomeric forms, prodrugs,
or pharmaceutically acceptable salt forms thereof, wherein:
R2 is selected from
H, halo, -OR11, -NO2, -CN, -CF3, -CHF2, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4
haloalkyl, and C1-C4 alkyl;
R5 is H, methyl or a prodrug of an amino group;
R6 is selected from H;
C1-C6 alkyl substituted by 0-2 R22;
C2-C6 alkenyl substituted by 0-2 R22;
C2-C6 alkynyl substituted by 0-2 R22; and
C3-C7 cycloalkyl substituted by 0-2 R22;
X is H,-NR9R10, halo, OR12, C1-C4 alkyl, or C2-C4 alkenyl;
R9 and R10 at each occurrence are each independently selected from H, -NH2;
C1-C6 alkyl substituted by 0-1 R19;
C2-C6 alkenyl substituted by 0-1 R19;
C2-C6 alkynyl substituted by 0-1 R19;
C6-C10 aryl substituted by 0-5 R19;
C3-C7 carbocyclyl substituted by 0-5 R19; and
5 to 14 membered heterocyclyl group substituted by 0-5 R19, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R19, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N,
O, and S;
alternatively, R9 and R10, together with the nitrogen to which they are attached, form a
3-7 membered heterocyclic ring, wherein said 3-7 membered heterocyclic ring
contains a nitrogen atom and optionally a second atom selected from N, O, S,
S(=O), and S(=O)2, whenein said 3-7 membered heterocyclic ring is substituted
with 0-1 R17;
R11 at each occurrence is independently selected from H, C1-C4 alkyl, and C1-C4
haloalkyl;
R12 at each occurrence is independently selected from H, C1-C4 haloalkyl and C1-C4
alkyl substituted with 0-1 R19;
R17 is H or C1-C4 alkyl substitutec: by 0-1 R19;
R19 at each occurrence is independently selected from H, -NR23R24, halo, -NO2, -CN, -
CF3, C1-C4 haloalkyl, -NHOH, OR25, C(=O)R25, C(=O)OR25, OC(=O)R25,
C(=O)NR23R24, NHC(=O)R25, NHCO2R25, OC(=O)NR23R24, NHC(=S)R25,
SR25, S(=O)R25, S(=O)2R25; S(=O)2NR23R24, -NHS(=O)2R25,
C1-C4 alkyl substituted by 0-1 R30;
C2-C4 alkenyl substituted by 0-1 R30;
C2-C4 alkynyl substituted by 0-1 R30;
C6-C10 aryl substituted by 0-5 R30;
C3-C7 carbocyclyl substitute by 0-5 R30; and
5 to 14 membered heterocyolyl group substituted by 0-5 R30, wherein said
heterocyclyl group comprises one, two, or three heteroatoms selected from
N, O, and S;
5 to 14 membered heteroaryl group substituted by 0-5 R30, wherein said
heteroaryl group comprises one, two, or three heteroatoms selected from N;
O, and S;
R22 is H, -NR23R24, -N3, halo, -NO2 -CN, -CF3, C1-C4 alkyl, C2-C4 alkenyl, C2-C4
alkynyl, C1-C4 haloalkyl, C3-C7carbocyclyl, phenyl, -NHOH, OR25, C(=O)R25,
C(=O)OR25, OC(=O)R25, C(=O)NR23R24, NHC(=O)R25, NHCO2R25,
OC(=O)NR23R24, NHC(=S)R25, SR25, S(=O)R25, S(=O)2R25; S(=O)2NR23R24. or
-NHS(=O)2R25;
R23 and R24 at each occurrence are each independently selected from H or C1-C4 alkyl;
R25 at each occurrence is each independently selected from H or C1-C4 alkyl; and
R30 is H, F, Cl, Br, -CF3, C1-C4 alkyl, and C1-C4 alkoxy.
17. The compound of Claim 9 wherein R6 is cyclobutyl, cyclopentyl, or eye
18. The compound of Claim 9 wherein R6 is cyclopentyl.
19. The compound of Claim 9 wherein X is -NR9r10.
20. The compound of Claim 9 wherein R6 is cyclopentyl and X is -NR9R10.
21. The compound of Claim 1 of Formula (H)
and stereoisomers forms, mixtures; of stereoisomeric forms, tautomeric forms or
pharmaceutically acceptable salt forms thereof, wherein:
R1, R2, R3, and R4 at each occurrence are independently selected from H, F, Cl, Br, -
OCH3, -NO2, -CN, and -CF-;
R5 is H;
R6 is selected from H, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, n-pentyl, i-
pentyl, allyl, cyclopentyl, cyclohexyl,
-CH2CH2OCH3, -CH2CH2OCH2CH3, -CH2CH2CH2N3, and -
CH2CH2CH2NHCH3;
R7 is H or -NO2;
X is selected from H, Cl, methyl, ethyl, propyl, butyl,
-OH; -OCH2CH2N(CH3)2; -OCH2CH2(pyrid-3-yl);
-NHCH3; -NCH2CH3; -NHCH(CH3)2; -NHCH2CH2CH2CH3; -
NHCH2CH(CH3)2;
-NHCH2CH2CF3; -NHCH=CH2; -NHCH2CH=CH2;
-NHCH2CH2N(CH3)2; -N( CH3)CH2CH2N(CH3)2; -NHCH2CH2CH2N(CH3)2;
-NHCH2CH2CH2NH(CH3);
-NHCH2CH2NH2; -NHCH2CH2CH2NH2; -N(H)CH2CH(NH2)CH3;
-N(CH3)CH2CH2N(CH2CH3)2; -NHNH2; -NHCH2CH2NHC(=O)CH3;
-N(CH2CH2OCH3)2; -N(H)CH2CH2OCH3; -N(H)CH2CH2CH2OCH3;
-N(H)CH2CH2OCH2CH3; -N(H)CH2CH2OCH2CH2CH3;
-N(CH2CH2OH)2; -N(H)CH2CH(OH)CH3; -N(H)CH2CH(OH)CH2CH3;
-N(H)CH2CH(OH)CH2OH;
-NH(pyrid-3-yl); -NH(4-F-pyrid-3-yl); -NH(4-MeO-pyrid-3-yl); piperazin-1-yl;
22. A compound selected from one of the following:
3-(2-Cyclopen1yl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-
carbonitrile;
5-Bromo-3-(2-clopentyl-2H-pyrazolo[3,4-d]piyrimidin-4-yl)-2-oxo-1,3-dihydro-indol-2-one;
6-Chloro-3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-1,3-dihydro-indol-2-one;
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-nitro-1,3-dihydro-indol-2-one;
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidm-4-yl)-5-trifluoromethyl-1,3-dihydro-indol-2-one;
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5,7-difluoro-1,3-dihydro-indol-2-one;
5-Chloro-3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-1,3-dihydro-indol-2-one;
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidm-4-yl)-1,3-dihydro-indo-2-one;
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-7-
carbonhrile;
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrmiidm-4-yl)-2-oxo-2,3-dihydro-1H-indole-7-
carbonitrile;
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-5-fluoro-1,3-dihydro-indol-2-one;
3-(2-Cyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-6-fluoro-1,3-dihydro-indol-2-one;
3-(2-Ctyclopentyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-4,5-difluoro-1,3-dihydro-indol-2-one;
5-Chloro-3-(2-isopropyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-1,3-dihydro-indol-2-one; and
3-(2-Isopropyl-2H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-oxo-2,3-dihydro-1H-indole-5-carbonitrile
and pharrnaceutically acceptable salt forms thereof.
23. A pharmaceutical compositions comprising a compound according to one of Claims 1 to
22 and one or more pharmaceutically acceptable excipients.
24. A method of treating a chronic neurodegenerative disease comprising administering to a
mammal a therapeutically effective amount of a compound according to one of Claims 1-22,
wherein said chronic neurodegenerative diseases is selected from Alzheimer's Disease,
Parkinson's Disease, progressive supranuclear palsy, subacute panencephalitic parkinsonism,
postencephalitic parkinsonism, dementia puglistica, guan-parkinsonial dementia complex, Pick's
disease, corticobasal degeneration,
frontotemporal dementia with parkinsonism, Huntington's disease, AIDS associated
dementia, amyotrophic lateral sclerosis;, and multiple sclerosis.
25. A method of treating a neurotraumatic disease comprising administering to a
mammal a therapeutically effective amount of a compound according to one of Claims
1-22, wherein said neurotraumatic disease is selected from acute stroke, mood disorders
such as schizophrenia and bipolar disorders, promotion of functional recovery post
stroke, cerebral bleeding (solitary cerebral amyloid anigopathy), hair loss, obesity,
atherosclerotic cardiovascular disease, hypertension, polycystic ovary syndrome,
syndrome X, ischemia, traumatic brain injury, cancer, leukopenia, Down's syndrome,
Lewy body disease, inflammation and immunodeficiency.
26. A method, of treating depression comprising administering to a mammal a
therapeutically effective amount of a compound according to one of Claims 1-22.
27. A method of treating diabetes wmprismg adrninistering to a mammal a
therapeutically effective amount of a compound according to one of Claims 1-22.

The present invention is related to chemical compositions, processes for the
preparation thereof and uses of the composition. Particularly, the present
invention relates to compositions that include substituted heterobicyclic
pyrimidines of Formula (I): wherein R, R, R,
R, R, X, W, and ring A are as defined herein; pharmaceutical
compositions of substituted heterobicyclic pyrimidines of Formula (I); and their
use in the treatment of chronic neurodegenerative diseases, neurotraumatic
diseases, depression and/or diabetes. More particularly, the present invention
relates to substituted pyrazolopyrimidines of Formula (I).

Documents:

5003-kolnp-2008-abstract.pdf

5003-KOLNP-2008-ASSIGNMENT.pdf

5003-kolnp-2008-claims.pdf

5003-KOLNP-2008-CORRESPONDENCE-1.1.pdf

5003-kolnp-2008-correspondence.pdf

5003-kolnp-2008-description (complete).pdf

5003-kolnp-2008-form 1.pdf

5003-kolnp-2008-form 2.pdf

5003-kolnp-2008-form 3.pdf

5003-kolnp-2008-form 5.pdf

5003-KOLNP-2008-GPA.pdf

5003-kolnp-2008-international publication.pdf

5003-kolnp-2008-international search report.pdf

5003-KOLNP-2008-OTHERS.pdf

5003-kolnp-2008-pct request form.pdf

5003-kolnp-2008-specification.pdf

abstract-5003-kolnp-2008.jpg


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Patent Number 263235
Indian Patent Application Number 5003/KOLNP/2008
PG Journal Number 42/2014
Publication Date 17-Oct-2014
Grant Date 15-Oct-2014
Date of Filing 10-Dec-2008
Name of Patentee CEPHALON, INC.
Applicant Address 41 MOORES ROAD, P.O. BOX 4011, FRAZER, PENNSYLVANIA
Inventors:
# Inventor's Name Inventor's Address
1 THOMAS R. BAILEY 217 AMANDA LANE PHOENIXVILLE, PENNSYLVANIA 19460 US
2 EDWARD R. BACON 1006 SKYLINE CIRCLE AUDUBON, PENNSYLVANIA 19403
3 NADINE C. BECKNELL 1537 BIRDELL ROAD COATESVILLE, PENNSYLVANIA 19320
4 DIANE E. GINGRICH 23 SOUTHWIND LANE DOWNINGTOWN, PENNSYLVANIA 19335
5 GREG HOSTETLER 7 SHELDRAKE ROAD NEWARK, DELAWARE 19713
6 ROBERT L. HUDKINS 430 S. SADDLEBROOK CIRCLE CHESTER SPRINGS, PENNSYLVANIA 19425
7 KEITH S. LEARN 3039 GOTTSCHALL ROAD PERKIOMENVILLE, PENNSYLVANIA 18074
8 JASON C. WAGNER 720 BARON CREST WAY COATESVILLE, PENNSYLVANIA 19320
PCT International Classification Number A61K31/5377; A61K31/496; A61K31/517
PCT International Application Number PCT/US2007/011619
PCT International Filing date 2007-05-15
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 11/803,320 2007-05-14 U.S.A.
2 60/800,375 2006-05-15 U.S.A.