U.S. patent application number 15/337237 was filed with the patent office on 2017-05-04 for pyrrolo-, pyrazolo-, imidazo-pyrimidine and pyridine compounds that inhibit mnk1 and mnk2.
The applicant listed for this patent is EFFECTOR THERAPEUTICS, INC.. Invention is credited to Justin T. Ernst, Siegfried H. Reich, Paul A. Sprengeler, Stephen E. Webber.
Application Number | 20170121339 15/337237 |
Document ID | / |
Family ID | 57256471 |
Filed Date | 2017-05-04 |
United States Patent
Application |
20170121339 |
Kind Code |
A1 |
Sprengeler; Paul A. ; et
al. |
May 4, 2017 |
PYRROLO-, PYRAZOLO-, IMIDAZO-PYRIMIDINE AND PYRIDINE COMPOUNDS THAT
INHIBIT MNK1 AND MNK2
Abstract
The present invention provides synthesis, pharmaceutically
acceptable formulations and uses of compounds in accordance with
Formula I, or a stereoisomer, tautomer or pharmaceutically
acceptable salt thereof. ##STR00001## For Formula I compounds
A.sup.1, A.sup.2, A.sup.3, A.sup.4, A.sup.5, A.sup.6, A.sup.7,
W.sup.1, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5a, R.sup.5b,
R.sup.6, R.sup.7, R.sup.7a, R.sup.7b, R.sup.8, R.sup.8a, R.sup.8b,
R.sup.9, R.sup.9a, R.sup.9b and R.sup.10 and subscripts "m" and "n"
are as defined in the specification. The inventive Formula I
compounds are inhibitors of Mnk and find utility in any number of
therapeutic applications, including but not limited to treatment of
inflammation and various cancers.
Inventors: |
Sprengeler; Paul A.;
(Escondido, CA) ; Reich; Siegfried H.; (La Jolla,
CA) ; Ernst; Justin T.; (San Diego, CA) ;
Webber; Stephen E.; (San Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
EFFECTOR THERAPEUTICS, INC. |
San Diego |
CA |
US |
|
|
Family ID: |
57256471 |
Appl. No.: |
15/337237 |
Filed: |
October 28, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62247966 |
Oct 29, 2015 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 43/00 20180101;
A61P 35/02 20180101; C07D 487/04 20130101; C07D 471/04 20130101;
A61P 35/00 20180101; A61P 25/00 20180101; C07D 519/00 20130101 |
International
Class: |
C07D 487/04 20060101
C07D487/04; C07D 519/00 20060101 C07D519/00 |
Claims
1. A compound according to Formula (I): ##STR00058## a
stereoisomer, a tautomer or a pharmaceutically acceptable salt
thereof wherein: A.sup.1 and A.sup.2 independently are --N-- or
--CR.sup.5a; A.sup.3 is --N-- or --CR.sup.6; A.sup.4 is --N-- or
--CR.sup.5b; A.sup.5 is --NR' or --CR.sup.7aR.sup.7b; A.sup.6 and
A.sup.7 are independently --N-- or --CR.sup.8a when represents a
bond, otherwise A.sup.6 and A.sup.7 are independently --NR.sup.8 or
--CR.sup.8aR.sup.8b; W.sup.1 is O, S, NH, NO(R.sup.9) or
CR.sup.9aR.sup.9b; m and n independently are 1, 2 or 3; R.sup.1 and
R.sup.2 independently are --H, --NHR.sup.10, NHR.sup.10-alkylene,
(C.sub.1-C.sub.8)alkyl, (C.sub.2-C.sub.8)alkenyl,
(C.sub.2-C.sub.8)alkynyl, cycloalkyl, heterocyclyl, heteroaryl,
aryl, arylalkylene, cycloalkylalkylene, heterocyclylalkylene or
heteroarylalkylene; or R.sup.1 and R.sup.2 together with the carbon
atom to which they are attached form a cycloalkyl or heterocyclyl
ring; R.sup.3 and R.sup.4 independently are --H, --OH, --CN,
--SR.sup.10, S(O).sub.2(C.sub.1-C.sub.8) alkyl, --C(O)NHR.sup.10,
--C(O)NR.sup.10R.sup.10, --NHR.sup.10, --NR.sup.10R.sup.10,
NHR.sup.10-alkylene, NR.sup.10R.sup.10-alkylene,
(C.sub.1-C.sub.8)alkyl, (C.sub.2-C.sub.8)alkenyl,
(C.sub.2-C.sub.8)alkynyl, (C.sub.1-C.sub.8)haloalkyl,
--O(C.sub.1-C.sub.8)alkyl, --O(C.sub.1-C.sub.8)haloalkyl,
--O(C.sub.1-C.sub.8)alkyleneNHR.sup.10,
--O(C.sub.1-C.sub.8)alkyleneNR.sup.10R.sup.10, cycloalkyl,
heterocyclyl, heteroaryl, aryl, arylalkylene, cycloalkylalkylene,
heterocyclylalkylene, heteroarylalkylene, alkylaminyl,
alkylcarbonylaminyl, cycloalkylcarbonylaminyl, cycloalkylaminyl, or
heterocyclylaminyl; R.sup.5a is --H, --OH, halogen, --CN, acetyl,
--(C.sub.1-C.sub.8)alkyl, --S(C.sub.1-C.sub.8)alkyl,
--(C.sub.2-C.sub.8)alkenyl, --(C.sub.2-C.sub.8)alkynyl,
--O(C.sub.1-C.sub.8)alkyl, (C.sub.1-C.sub.8)haloalkyl,
--NHR.sup.10, --NR.sup.10R.sup.10, NHR.sup.10-alkylene,
NR.sup.10R.sup.10-alkylene, or --O(C.sub.1-C.sub.8)haloalkyl;
R.sup.5b and R.sup.6 is --H, --OH, --SH, --CN,
--S(O).sub.2R.sup.10, halogen, --S(C.sub.1-C.sub.8)alkyl,
--NHR.sup.10, --NR.sup.10R.sup.10, (C.sub.1-C.sub.8)alkyl,
(C.sub.2-C.sub.8)alkenyl, (C.sub.2-C.sub.8)alkynyl,
(C.sub.1-C.sub.8)haloalkyl, --O(C.sub.1-C.sub.8)haloalkyl,
--O(C.sub.1-C.sub.8)alkyl, --O(C.sub.1-C.sub.8)alkyleneNHR.sup.10,
--O(C.sub.1-C.sub.8)alkyleneNR.sup.10R.sup.10,
--(C.sub.1-C.sub.8)alkyleneNHR.sup.10,
--(C.sub.1-C.sub.8)alkyleneNR.sup.10R.sup.10,
--S(C.sub.1-C.sub.8)alkyl, cycloalkyl, heterocyclyl, heteroaryl, or
aryl; R.sup.7 is --H, --OH, acetyl, --(C.sub.1-C.sub.8)alkyl,
--C(O)alkyl, --C(O)cycloalkyl, --C(O)O--(C.sub.1-C.sub.8)alkyl,
cycloalkyl, aryl, heteroaryl or heterocyclyl; R.sup.7a and R.sup.7b
independently are --H, --OH, acetyl, --(C.sub.1-C.sub.8)alkyl,
--O(C.sub.1-C.sub.8)alkyl, --C(O)alkyl, --C(O)cycloalkyl,
--C(O)O--(C.sub.1-C.sub.8)alkyl, cycloalkyl, aryl, heteroaryl or
heterocyclyl; R.sup.8 is --H, --OH, acetyl, (C.sub.1-C.sub.8)alkyl,
cycloalkyl, heterocyclyl, heteroaryl or aryl; R.sup.8a and R.sup.8b
independently are --H, --OH, --CN, acetyl, --SH,
--S(O).sub.2R.sup.10, halogen, --S(C.sub.1-C.sub.8)alkyl,
--NHR.sup.10, --NR.sup.10R.sup.10, (C.sub.1-C.sub.8)alkyl,
(C.sub.1-C.sub.8)haloalkyl, --O(C.sub.1-C.sub.8)alkyl,
--O(C.sub.1-C.sub.8)alkylNHR.sup.10,
--O(C.sub.1-C.sub.8)alkylNR.sup.10R.sup.10,
--(C.sub.1-C.sub.8)alkylNHR.sup.10,
--(C.sub.1-C.sub.8)alkylNR.sup.10R.sup.10, cycloalkyl,
heterocyclyl, heteroaryl or aryl; R.sup.9, R.sup.9a and R.sup.9b
are independently --H, (C.sub.1-C.sub.8)alkyl,
(C.sub.2-C.sub.8)alkenyl, (C.sub.2-C.sub.8)alkynyl, cycloalkyl,
heterocyclyl, heteroaryl, aryl, arylalkylene, cycloalkylalkylene,
heterocyclylalkylene, or heteroarylalkylene, or R.sup.9a and
R.sup.9b together with the carbon atom to which they are attached
form a cycloalkyl or heterocyclyl ring; R.sup.10 is --H, --OH,
--C(O)O(C.sub.1-C.sub.8)alkyl, --C(O)(C.sub.1-C.sub.8)alkyl,
--C(O)--NH.sub.2, --C(O)--NH(C.sub.1-C.sub.8)alkyl,
NH.sub.2--C(O)-alkylene, --S(C.sub.1-C.sub.8)alkyl, acetyl,
--(C.sub.1-C.sub.8)alkyl, (C.sub.2-C.sub.8)alkenyl,
(C.sub.2-C.sub.8)alkynyl, --O(C.sub.1-C.sub.8)alkyl,
(C.sub.1-C.sub.8) haloalkyl, alkylcarbonylaminyl, alkylaminyl,
--C(O)alkyl, --C(O)cycloalkyl, --C(O)O--(C.sub.1-C.sub.8)alkyl,
aryl, heteroaryl, heterocyclyl or cycloalkyl; wherein any alkyl,
cycloalkyl, heterocyclyl, heteroaryl, aryl, arylalkylene,
cycloalkylalkylene, heterocyclylalkylene, heteroarylalkylene,
alkylaminyl, alkylcarbonylaminyl, cycloalkylcarbonylaminyl,
cycloalkylaminyl or heterocyclylaminyl is optionally substituted
with 1, 2 or 3 groups selected from --OH, --CN, --SH,
--S(O)NH.sub.2, --S(O)NH.sub.2, halogen, --NH.sub.2,
--NH(C.sub.1-C.sub.4)alkyl, --N[(C.sub.1-C.sub.4)alkyl].sub.2,
--C(O)NH.sub.2, --COOH, --COOMe, acetyl, --(C.sub.1-C.sub.8)alkyl,
--O(C.sub.1-C.sub.8)alkyl (C.sub.2-C.sub.8)alkenyl,
(C.sub.2-C.sub.8)alkynyl, haloalkyl, thioalkyl, cyanomethylene,
alkylaminyl, NH.sub.2--C(O)-alkylene, NH.sub.2--C(O)-alkylene,
--NH(Me)-C(O)-alkylene, --CH.sub.2--C(O)-lower alkyl, --C(O)-lower
alkyl, alkylcarbonylaminyl, cycloalkyl, cycloalkylalkylene,
cycloalkylalkenylene, cycloalkylcarbonylaminyl, cycloalkylaminyl,
--CH.sub.2--C(O)-cycloalkyl, --C(O)-cycloalkyl,
--CH.sub.2--C(O)-aryl, --CH.sub.2-aryl, --C(O)-aryl,
--CH.sub.2--C(O)-heterocycloalkyl, --C(O)-heterocycloalkyl,
heterocyclylaminyl or heterocyclyl; and --- represents the option
of having a double bond.
2. The compound according to claim 1 wherein "m" and "n" are both
1, A.sup.5 is --NR.sup.7, W.sup.1 is O and R.sup.7 is --H.
3. The compound according to claim 1 wherein R.sup.1 and R.sup.2
independently are --H, methyl, ethyl, i-propyl, --NH.sub.2,
aminomethylene or --CH.sub.3O--C(O)NH-methylene.
4. The compound according to claim 1 wherein R.sup.1 and R.sup.2
together with the carbon atom to which they are attached form a
cycloalkyl ring selected from the group consisting of cyclobutyl,
cyclopentyl, cyclohexyl, 1-(2,2-difluoroethyl)piperidine or
1-methylpiperidine.
5. The compound according to claim 1 wherein A.sup.2 is
--CR.sup.5a, A.sup.3 is --CR.sup.6 and A.sup.4 is --CR.sup.5b.
6. The compound according to claim 5 wherein R.sup.5a and R.sup.6
independently are --H, chlorine, fluorine, or methyl and R.sup.5b
is --H.
7. The compound according to claim 1 wherein A.sup.2 is
--CR.sup.5a, A.sup.3 is --CR.sup.6, A.sup.4 is --N and R.sup.5a and
R.sup.6 independently are --H, chlorine, fluorine or methyl.
8. The compound according to claim 1 wherein A.sup.2 is --N,
A.sup.3 is --CR.sup.6 and A.sup.4 is CR.sup.5b.
9. The compound according to claim 8 wherein R.sup.6 is --C(Me) or
--CH and R.sup.5b is --H.
10. The compound according to claim 1 wherein R.sup.3 and R.sup.4
independently are --H.
11. The compound according to claim 1 wherein R.sup.3 is --H and
R.sup.4 is chlorine, fluorine, methyl, ethyl or --NH.sub.2.
12. The compound according to claim 1 wherein A.sup.6 and A.sup.7
are --CR.sup.8a --- represents a bond, and R.sup.8a is --H,
heteroaryl or aryl.
13. The compound according to claim 12 wherein A.sup.6 and A.sup.7
are --CH.
14. The compound according to claim 12 wherein A.sup.6 is --CH and
A.sup.7 is --C(heteroaryl) or --C(aryl).
15. The compound according to claim 1 selected from the following
table: TABLE-US-00003 ##STR00059## ##STR00060## ##STR00061##
##STR00062## ##STR00063## ##STR00064## ##STR00065## ##STR00066##
##STR00067## ##STR00068## ##STR00069## ##STR00070## ##STR00071##
##STR00072## ##STR00073## ##STR00074## ##STR00075##
##STR00076##
16. A pharmaceutical composition comprising (i) a therapeutically
effective amount of at least one compound according to claim 1 or a
stereoisomer, a tautomer or a pharmaceutically acceptable salt
thereof; (ii) in combination with a pharmaceutically acceptable
carrier, diluent or excipient.
17. A method for attenuating or inhibiting the activity of MnK in
at least one cell overexpressing Mnk comprising contacting the at
least one cell with a compound according to claim 1 or a
stereoisomer, tautomer or pharmaceutically acceptable salt
thereof.
18. The method of claim 17 wherein the at least one cell is a colon
cancer cell, a gastric cancer cell, a thyroid cancer cell, a lung
cancer cell, a leukemia cell, a B-cell lymphoma, a T-cell lymphoma,
a hairy cell lymphoma, Hodgkin's lymphoma cell, non-Hodgkin's
lymphoma cell, Burkitt's lymphoma cell, a pancreatic cancer cell, a
melanoma cell, a multiple melanoma cell, a brain cancer cell, a CNS
cancer cell, a renal cancer cell, a prostate cancer cell, an
ovarian cancer cell, or a breast cancer cell.
19. A method for treating a Mnk dependent condition in a mammal in
need thereof comprising administering to the mammal (i) a
therapeutically effective amount of at least one compound according
to claim 1 or a stereoisomer, tautomer or pharmaceutically
acceptable salt thereof, or (ii) a pharmaceutical composition of
claim 16.
20. The method of claim 19 wherein the Mnk dependent condition is
colon cancer, colorectal cancer, gastric cancer, thyroid cancer,
lung cancer, leukemia, B-cell lymphoma, T-cell lymphoma, hairy cell
lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's
lymphoma, pancreatic cancer, melanoma, multiple melanoma, brain
cancer, CNS cancer, renal cancer, prostate cancer, ovarian cancer,
breast cancer, Alzheimer's, Parkinson's, Fragile X Syndrome and
autism disorders.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Patent Application No. 62/247,966, filed Oct. 29, 2015, which is
incorporated herein by reference in its entirety.
FIELD
[0002] The present invention generally relates to compounds having
activity as inhibitors of MAP kinase-interacting kinase (Mnk), for
example Mnk1 and Mnk2, as well as to related compositions and
methods for utilizing the inventive compounds as therapeutic agents
for treatment of Mnk dependent diseases, including the treatment of
cancer.
BACKGROUND
[0003] Eukaryotic initiation factor 4E (eIF4E) is a general
translation factor but it has the potential to enhance
preferentially the translation of messenger RNAs (mRNAs) that lead
to production of malignancy-associated proteins. This selectivity
may relate to an increased requirement for eIF4E and its binding
partners for the translation of mRNAs containing extensive
secondary structure in their 5'-untranslated regions (5'-UTRs).
These mRNAs include those encoding certain proteins that control
cell cycle progression and tumorigenesis. Under normal cellular
conditions the translation of these malignancy-associated mRNAs is
suppressed as the availability of active eIF4E is limited; however,
their levels can increase when eIF4E is over-expressed or
hyperactivated. Elevated levels of eIF4E have been found in many
types of tumors and cancer cell lines including cancers of the
colon, breast, bladder, lung, prostate, gastrointestinal tract,
head and neck, Hodgkin's lymphomas and neuroblastomas.
[0004] Initiation of cap-dependent translation is thought to depend
on the assembly of eIF4F, an initiation factor complex including
eIF4E, the scaffold protein eIF4G, and the RNA helicase eIF4A.
Because eIF4E is the only one of these proteins that binds directly
to the mRNA cap structure, it is the key factor for the assembly of
eIF4F at the 5' cap. The scaffold protein, eIF4G, also recruits the
40S ribosomal subunit to the mRNA via its interaction with eIF3 and
binds eIF4B, a protein that aids the RNA-helicase function of
eIF4A, thus facilitating the translation of mRNAs that contain
structured 5'-UTRs. The availability of eIF4E as part of the eIF4F
complex is a limiting factor in controlling the rate of
translation, and therefore eIF4E is an important regulator of mRNA
translation.
[0005] Regulation of eIF4E activity forms a node of convergence of
the PI3K/Akt/mTOR and Ras/Raf/MAPK signaling pathways. The PI3K
(phosphoinositide 3-kinase)/PTEN (phosphatase and tensin homologue
deleted on chromosome ten)/Akt/mTOR (mammalian target of rapamycin)
pathway is often involved in tumorgenesis and in sensitivity and
resistance to cancer therapy. Deregulated signaling through the
PI3K/PTEN/Akt/mTOR pathway is often the result of genetic
alterations in critical components of this pathway and/or mutations
at upstream growth factor receptors or signaling components. PI3K
initiates a cascade of events when activated by, for example,
extracellular growth factors, mitogens, cytokines and/or receptors,
PDK1 activates Akt, which in turn phosphorylates and inactivates
the tumor suppressor complex comprising TSC1 and 2 (tuberous
sclerosis complex 1/2), resulting in the activation of mTORC1
(target of rapamycin complex 1) by Rheb-GTP. Activation of PDK1 and
Akt by PI3Ks is negatively regulated by PTEN.
[0006] PTEN is a critical tumor suppressor gene and is often
mutated or silenced in human cancers. Its loss results in
activation of Akt and increases downstream mTORC1 signaling. The
involvement of mTOR complex1 (mTORC1) in neoplastic transformation
appears to depend on its regulatory role toward the eIF4F complex;
overexpression of eIF4E can confer resistance to rapamycin. mTORC1
regulates the eIF4F complex assembly that is critical for the
translation of mRNAs associated with cell growth, prevention of
apoptosis and transformation. mTORC1 achieves this by
phosphorylation and inactivation of 4E-BPs and the subsequent
dissociation of 4E-BPs from eIF4E. This then enables eIF4E to
interact with the scaffold protein eIF4G, permitting assembly of
the eIF4F complex for the translation of structured mRNAs. mTORC1
also promotes activation of the translational activator, S6K, which
phosphorylates the ribosomal protein S6 and other substrates,
including eIF4B. mTORC1 signaling is inhibited by rapamycin and its
analogues (rapalogs), although these compounds act allosterically,
rather than directly inhibiting mTOR kinase activity.
[0007] Given the importance of the PI3K/Akt/mTOR pathway in
regulating mRNA translation of genes that encode for pro-oncogenic
proteins and activated mTORC1 signaling in a high proportion of
cancers, these kinases have been actively pursued as oncology drug
targets. A number of pharmacological inhibitors have been
identified, some of which have reached advanced clinical stages.
However, it has recently become clear that the mTOR pathway
participates in a complicated feedback loop that can impair
activation of Akt. It has been shown that prolonged treatment of
cancer cells or patients with mTOR inhibitors causes elevated PI3K
activity that leads to phosphorylation of Akt and eIF4E, and
promotes cancer cell survival. eIF4E, acting downstream of Akt and
mTOR, recapitulates Akt's action in tumorigenesis and drug
resistance, and Akt signaling via eIF4E is an important mechanism
of oncogenesis and drug resistance in vivo.
[0008] In addition to the PI3K/Akt/mTOR pathway, eIF4E is also the
target of the Ras/Raf/MAP signaling cascade which is activated by
growth factors and for the stress-activated p38 MAP kinase pathway.
Erk1/2 and p38 then phosphorylate MAP kinase-interacting kinase 1
(Mnk1) and MAP kinase-interacting kinase 2 (Mnk2). The Erk pathway
is also activated in many cancers, reflecting, for example,
activating mutations in Ras (found in around 20% of tumors) or loss
of function of the Ras GTPase-activator protein NF1. Mnk1 and Mnk2
are threonine/serine protein kinases and specifically phosphorylate
serine 209 (Ser209) of eIF4E within the eIF4F complex, by virtue of
the interaction between eIF4E and the Mnks, which serves to recruit
Mnks to act on eIF4E. Mice with mutated eIF4E, in which Ser209 is
replaced by alanine, shows no eIF4E phosphorylation and
significantly attenuated tumor growth. Significantly, while Mnk
activity is necessary for eIF4E-mediated oncogenic transformation,
it is dispensable for normal development. Pharmacologically
inhibiting Mnks thus presents an attractive therapeutic strategy
for cancer.
[0009] Despite increased understanding of Mnk structure and
function, little progress has been made with regard to the
discovery of pharmacological Mnk inhibitors and relatively few Mnk
inhibitors have been reported: CGP052088 (Tschopp et al., Mol Cell
Biol Res Commun. 3(4):205-211, 2000); CGP57380 (Rowlett et al., Am
J Physiol Gastrointest Liver Physiol. 294(2):G452-459, 2008); and
Cercosporamide (Konicek et al., Cancer Res. 71(5):1849-1857, 2011).
These compounds, however, have mainly been used for the purpose of
Mnk target validation. More recently, investigators have proposed
further compounds for treating diseases influenced by the
inhibition of kinase activity of Mnk1 and/or Mnk2, including, for
example, the compounds disclosed in WO 2014/044691 and the various
patent documents cited therein and the
4-(dihydropyridinon-3-yl)amino-5-methylthieno[2,3,-d]pyrimidines
disclosed by Yu et al., European Journal of Med. Chem., 95:
116-126, 2015).
[0010] Accordingly, while advances have been made in this field
there remains a significant need in the art for compounds that
specifically inhibit Mnk kinase activity, particularly with regard
to Mnk's role in regulation of cancer pathways, as well as for
associated composition and methods. The present invention fulfills
this need and provides further related advantages.
SUMMARY
[0011] The present invention is directed to compounds that inhibit
or modulate the activity of Mnk, as well as stereoisomers,
tautomers and pharmaceutically acceptable salts of such compounds.
The invention also is directed to pharmaceutically acceptable
compositions containing such compounds and associated methods for
treating conditions that would benefit from Mnk inhibition, such as
cancer.
[0012] In one embodiment the invention is directed to compounds
according to Formula I as well as to a stereoisomer, tautomer or
pharmaceutically acceptable salt of such compounds, wherein
##STR00002##
[0013] A.sup.1 and A.sup.2 independently are --N-- or
--CR.sup.5a;
[0014] A.sup.3 is --N-- or --CR.sup.6;
[0015] A.sup.4 is --N-- or --CR.sup.5b;
[0016] A.sup.5 is --NR.sup.7 or --CR.sup.7aR.sup.7b;
[0017] A.sup.6 and A.sup.7 are independently --N-- or --CR.sup.8a
when --------- represents a bond, otherwise A.sup.6 and A.sup.7 are
independently --NR.sup.8 or --CR.sup.8aR.sup.8b;
[0018] W.sup.1 is O, S, NH, NO(R.sup.9) or CR.sup.9aR.sup.9b;
[0019] m and n independently are 1, 2 or 3;
[0020] R.sup.1 and R.sup.2 independently are --H, --NHR.sup.10,
NHR.sup.10-alkylene, (C.sub.1-C.sub.8)alkyl,
(C.sub.2-C.sub.8)alkenyl, (C.sub.2-C.sub.8)alkynyl, cycloalkyl,
heterocyclyl, heteroaryl, aryl, arylalkylene, cycloalkylalkylene,
heterocyclylalkylene or heteroarylalkylene; or
[0021] R.sup.1 and R.sup.2 together with the carbon atom to which
they are attached form a cycloalkyl or heterocyclyl ring;
[0022] R.sup.3 and R.sup.4 independently are --H, --OH, --CN,
--SR.sup.10, S(O).sub.2(C.sub.1-C.sub.8) alkyl, --C(O)NHR.sup.10,
--C(O)NR.sup.10R.sup.10, --NHR.sup.10, --NR.sup.10R.sup.10,
NHR.sup.10-alkylene, NR.sup.10R.sup.10-alkylene,
(C.sub.1-C.sub.8)alkyl, (C.sub.2-C.sub.8)alkenyl,
(C.sub.2-C.sub.8)alkynyl, (C.sub.1-C.sub.8)haloalkyl,
--O(C.sub.1-C.sub.8)alkyl, --O(C.sub.1-C.sub.8)haloalkyl,
--O(C.sub.1-C.sub.8)alkyleneNHR.sup.10,
--O(C.sub.1-C.sub.8)alkyleneNR.sup.10R.sup.10, cycloalkyl,
heterocyclyl, heteroaryl, aryl, arylalkylene, cycloalkylalkylene,
heterocyclylalkylene, heteroarylalkylene, alkylaminyl,
alkylcarbonylaminyl, cycloalkylcarbonylaminyl, cycloalkylaminyl, or
heterocyclylaminyl;
[0023] R.sup.5a is --H, --OH, halogen, --CN, acetyl,
--(C.sub.1-C.sub.8)alkyl, --S(C.sub.1-C.sub.8)alkyl,
--(C.sub.2-C.sub.8)alkenyl, --(C.sub.2-C.sub.8)alkynyl,
--O(C.sub.1-C.sub.8)alkyl, (C.sub.1-C.sub.8)haloalkyl,
--NHR.sup.10, --NR.sup.10R.sup.10, NHR.sup.10-alkylene,
NR.sup.10R.sup.10-alkylene, or --O(C.sub.1-C.sub.8)haloalkyl;
[0024] R.sup.5b and R.sup.6 is --H, --OH, --SH, --CN,
--S(O).sub.2R.sup.10, halogen, --S(C.sub.1-C.sub.8)alkyl,
--NHR.sup.10, --NR.sup.10R.sup.10, (C.sub.1-C.sub.8)alkyl,
(C.sub.2-C.sub.8)alkenyl, (C.sub.2-C.sub.8)alkynyl,
(C.sub.1-C.sub.8)haloalkyl, --O(C.sub.1-C.sub.8)haloalkyl,
--O(C.sub.1-C.sub.8)alkyl, --O(C.sub.1-C.sub.8)alkyleneNHR.sup.10,
--O(C.sub.1-C.sub.8)alkyleneNR.sup.10R.sup.10,
--(C.sub.1-C.sub.8)alkyleneNHR.sup.10,
--(C.sub.1-C.sub.8)alkyleneNR.sup.10R.sup.10,
--S(C.sub.1-C.sub.8)alkyl, cycloalkyl, heterocyclyl, heteroaryl, or
aryl;
[0025] R.sup.7 is --H, --OH, acetyl, --(C.sub.1-C.sub.8)alkyl,
--C(O)alkyl, --C(O)cycloalkyl, --C(O)O--(C.sub.1-C.sub.8)alkyl,
cycloalkyl, aryl, heteroaryl or heterocyclyl;
[0026] R.sup.7a and R.sup.7b independently are --H, --OH, acetyl,
--(C.sub.1-C.sub.8)alkyl, --O(C.sub.1-C.sub.8)alkyl, --C(O)alkyl,
--C(O)cycloalkyl, --C(O)O--(C.sub.1-C.sub.8)alkyl, cycloalkyl,
aryl, heteroaryl or heterocyclyl;
[0027] R.sup.8 is --H, --OH, acetyl, (C.sub.1-C.sub.8)alkyl,
cycloalkyl, heterocyclyl, heteroaryl or aryl;
[0028] R.sup.8a and R.sup.8b independently are --H, --OH, --CN,
acetyl, --SH, --S(O).sub.2R.sup.10, halogen,
--S(C.sub.1-C.sub.8)alkyl, --NHR.sup.10, --NR.sup.10R.sup.10,
(C.sub.1-C.sub.8)alkyl, (C.sub.1-C.sub.8)haloalkyl,
--O(C.sub.1-C.sub.8)alkyl, --O(C.sub.1-C.sub.8)alkylNHR.sup.10,
--O(C.sub.1-C.sub.8)alkylNR.sup.10R.sup.10,
--(C.sub.1-C.sub.8)alkylNHR.sup.10,
--(C.sub.1-C.sub.8)alkylNR.sup.10R.sup.10, cycloalkyl,
heterocyclyl, heteroaryl or aryl;
[0029] R.sup.9, R.sup.9a and R.sup.9b are independently --H,
(C.sub.1-C.sub.8)alkyl, (C.sub.2-C.sub.8)alkenyl,
(C.sub.2-C.sub.8)alkynyl, cycloalkyl, heterocyclyl, heteroaryl,
aryl, arylalkylene, cycloalkylalkylene, heterocyclylalkylene, or
heteroarylalkylene, or
[0030] R.sup.9a and R.sup.9b together with the carbon atom to which
they are attached form a cycloalkyl or heterocyclyl ring;
[0031] R.sup.10 is --H, --OH, --C(O)O(C.sub.1-C.sub.8)alkyl,
--C(O)(C.sub.1-C.sub.8)alkyl, --C(O)--NH.sub.2,
--C(O)--NH(C.sub.1-C.sub.8)alkyl, NH.sub.2--C(O)-alkylene,
--S(C.sub.1-C.sub.8)alkyl, acetyl, --(C.sub.1-C.sub.8)alkyl,
(C.sub.2-C.sub.8)alkenyl, (C.sub.2-C.sub.8)alkynyl,
--O(C.sub.1-C.sub.8)alkyl, (C.sub.1-C.sub.8) haloalkyl,
alkylcarbonylaminyl, alkylaminyl, --C(O)alkyl, --C(O)cycloalkyl,
--C(O)O--(C.sub.1-C.sub.8)alkyl, aryl, heteroaryl, heterocyclyl or
cycloalkyl;
[0032] wherein any alkyl, cycloalkyl, heterocyclyl, heteroaryl,
aryl, arylalkylene, cycloalkylalkylene, heterocyclylalkylene,
heteroarylalkylene, alkylaminyl, alkylcarbonylaminyl,
cycloalkylcarbonylaminyl, cycloalkylaminyl or heterocyclylaminyl is
optionally substituted with 1, 2 or 3 groups selected from --OH,
--CN, --SH, --S(O)NH.sub.2, --S(O)NH.sub.2, halogen, --NH.sub.2,
--NH(C.sub.1-C.sub.4)alkyl, --N[(C.sub.1-C.sub.4)alkyl].sub.2,
--C(O)NH.sub.2, --COOH, --COOMe, acetyl, --(C.sub.1-C.sub.8)alkyl,
--O(C.sub.1-C.sub.8)alkyl (C.sub.2-C.sub.8)alkenyl,
(C.sub.2-C.sub.8)alkynyl, haloalkyl, thioalkyl, cyanomethylene,
alkylaminyl, NH.sub.2--C(O)-alkylene, NH.sub.2--C(O)-alkylene,
--NH(Me)-C(O)-alkylene, --CH.sub.2--C(O)-lower alkyl, --C(O)-lower
alkyl, alkylcarbonylaminyl, cycloalkyl, cycloalkylalkylene,
cycloalkylalkenylene, cycloalkylcarbonylaminyl, cycloalkylaminyl,
--CH.sub.2--C(O)-cycloalkyl, --C(O)-cycloalkyl,
--CH.sub.2--C(O)-aryl, --CH.sub.2-aryl, --C(O)-aryl,
--CH.sub.2--C(O)-heterocycloalkyl, --C(O)-heterocycloalkyl,
heterocyclylaminyl or heterocyclyl; and
[0033] --- represents the option of having a double bond.
[0034] The present invention also provides a pharmaceutical
composition comprising (i) a therapeutically effective amount of at
least one compound according to Formula I or a stereoisomer, a
tautomer or a pharmaceutically acceptable salt thereof; (ii) in
combination with a pharmaceutically acceptable carrier, diluent or
excipient.
[0035] Also provided by the present invention is a method for
attenuating or inhibiting the activity of MnK in at least one cell
overexpressing Mnk, comprising contacting the at least one cell
with a compound according to claim 1 or a stereoisomer, tautomer or
pharmaceutically acceptable salt thereof.
[0036] According to the inventive method at least one cell is a
colon cancer cell, a gastric cancer cell, a thyroid cancer cell, a
lung cancer cell, a leukemia cell, a B-cell lymphoma, a T-cell
lymphoma, a hairy cell lymphoma, Hodgkin's lymphoma cell,
non-Hodgkin's lymphoma cell, Burkitt's lymphoma cell, a pancreatic
cancer cell, a melanoma cell, a multiple melanoma cell, a brain
cancer cell, a CNS cancer cell, a renal cancer cell, a prostate
cancer cell, an ovarian cancer cell, or a breast cancer cell.
[0037] According to yet another embodiment the invention provides a
method for treating a Mnk dependent condition in a mammal in need
thereof comprising administering to the mammal (i) a
therapeutically effective amount of at least one compound according
to claim 1 or a stereoisomer, tautomer or pharmaceutically
acceptable salt thereof, or (ii) a pharmaceutical composition in
accordance with the invention.
[0038] Compounds and pharmaceutically acceptable formulations in
accordance with the invention are useful for treating an Mnk
dependent condition such as colon cancer, gastric cancer, thyroid
cancer, lung cancer, leukemia, B-cell lymphoma, T-cell lymphoma,
hairy cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma,
Burkitt's lymphoma, pancreatic cancer, melanoma, multiple melanoma,
brain cancer, CNS cancer, renal cancer, prostate cancer, ovarian
cancer, or breast cancer.
[0039] The above embodiments and other aspects of the invention are
readily apparent in the detailed description that follows. Various
references are set forth herein which describe in more detail
certain background information, procedures, compounds and/or
compositions, and are each hereby incorporated by reference in
their entirety.
DETAILED DESCRIPTION
[0040] In the following description certain specific details are
set forth in order to provide a thorough understanding of various
embodiments of the invention. However, one skilled in the art will
understand that the invention may be practiced without these
details. Unless the context requires otherwise, throughout the
present specification and claims, the word "comprise" and
variations thereof, such as, "comprises" and "comprising" are to be
construed in an open, inclusive sense (i.e., as "including, but not
limited to").
[0041] Reference throughout this specification to "one embodiment"
or "an embodiment" means that a particular feature, structure or
characteristic described in connection with the embodiment is
included in at least one embodiment of the present invention. Thus,
the appearances of the phrases "in one embodiment" or "in an
embodiment" in various places throughout this specification are not
necessarily all referring to the same embodiment. Furthermore, the
particular features, structures, or characteristics may be combined
in any suitable manner in one or more embodiments.
DEFINITIONS
[0042] As used herein, and unless noted to the contrary, the
following terms and phrases have the meaning noted below.
[0043] "Amino" refers to the --NH.sub.2 substituent.
[0044] "Aminocarbonyl" refers to the --C(O)NH.sub.2
substituent.
[0045] "Carboxyl" refers to the --CO.sub.2H substituent.
[0046] "Carbonyl" refers to a --C(O)-- or --C(.dbd.O)-- group. Both
notations are used interchangeably within the specification.
[0047] "Cyano" refers to the --C.ident.N substituent.
[0048] "Cyanoalkylene" refers to the -(alkylene)C.ident.N
substituent.
[0049] "Acetyl" refers to the --C(O)CH.sub.3 substituent.
[0050] "Hydroxy" or "hydroxyl" refers to the --OH substituent.
[0051] "Hydroxyalkylene" refers to the -(alkylene)OH
substituent.
[0052] "Oxo" refers to a .dbd.O substituent.
[0053] "Thio" or "thiol" refer to a --SH substituent.
[0054] "Alkyl" refers to a saturated, straight or branched
hydrocarbon chain radical consisting solely of carbon and hydrogen
atoms, having from one to twelve carbon atoms (C.sub.1-C.sub.12
alkyl), from one to eight carbon atoms (C.sub.1-C.sub.8 alkyl) or
from one to six carbon atoms (C.sub.1-C.sub.6 alkyl), and which is
attached to the rest of the molecule by a single bond. Exemplary
alkyl groups include methyl, ethyl, n-propyl, 1-methylethyl
(iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl),
3-methylhexyl, 2-methylhexyl, and the like.
[0055] "Lower alkyl" has the same meaning as alkyl defined above
but having from one to four carbon atoms (C.sub.1-C.sub.4
alkyl).
[0056] "Alkenyl" refers to an unsaturated alkyl group having at
least one double bond and from two to twelve carbon atoms
(C.sub.2-C.sub.12 alkenyl), from two to eight carbon atoms
(C.sub.2-C.sub.8 alkenyl) or from two to six carbon atoms
(C.sub.2-C.sub.6 alkenyl), and which is attached to the rest of the
molecule by a single bond, e.g., ethenyl, propenyl, butenyl,
pentenyl, hexenyl, and the like.
[0057] "Alkynyl" refers to an unsaturated alkyl group having at
least one triple bond and from two to twelve carbon atoms
(C.sub.2-C.sub.12 alkynyl), from two to ten carbon atoms
(C.sub.2-C.sub.10 alkynyl) from two to eight carbon atoms
(C.sub.2-C.sub.8 alkynyl) or from two to six carbon atoms
(C.sub.2-C.sub.6 alkynyl), and which is attached to the rest of the
molecule by a single bond, e.g., ethynyl, propynyl, butynyl,
pentynyl, hexynyl, and the like.
[0058] "Alkylene" or "alkylene chain" refers to a straight or
branched divalent hydrocarbon (alkyl) chain linking the rest of the
molecule to a radical group, consisting solely of carbon and
hydrogen, respectively. Alkylenes can have from one to twelve
carbon atoms, e.g., methylene, ethylene, propylene, n-butylene, and
the like. The alkylene chain is attached to the rest of the
molecule through a single or double bond. The points of attachment
of the alkylene chain to the rest of the molecule can be through
one carbon or any two carbons within the chain. "Optionally
substituted alkylene" refers to alkylene or substituted
alkylene.
[0059] "Alkenylene" refers to divalent alkene. Examples of
alkenylene include without limitation, ethenylene (--CH.dbd.CH--)
and all stereoisomeric and conformational isomeric forms thereof.
"Substituted alkenylene" refers to divalent substituted alkene.
"Optionally substituted alkenylene" refers to alkenylene or
substituted alkenylene.
[0060] "Alkynylene" refers to divalent alkyne. Examples of
alkynylene include without limitation, ethynylene, propynylene.
"Substituted alkynylene" refers to divalent substituted alkyne.
[0061] "Alkoxy" refers to a radical of the formula --OR.sub.a where
R.sub.a is an alkyl having the indicated number of carbon atoms as
defined above. Examples of alkoxy groups include without limitation
--O-methyl (methoxy), --O-ethyl (ethoxy), --O-propyl (propoxy),
--O-isopropyl (iso propoxy) and the like.
[0062] "Alkylaminyl" refers to a radical of the formula --NHR.sub.a
or --NR.sub.aR.sub.a where each R.sub.a is, independently, an alkyl
radical having the indicated number of carbon atoms as defined
above.
[0063] "Cycloalkylaminyl" refers to a radical of the formula
--NHR.sub.a where R.sub.a is a cycloalkyl radical as defined
herein.
[0064] "Alkylcarbonylaminyl" refers to a radical of the formula
--NHC(O)R.sub.a, where R.sub.a is an alkyl radical having the
indicated number of carbon atoms as defined herein.
[0065] "Cycloalkylcarbonylaminyl" refers to a radical of the
formula --NHC(O)R.sub.a, where R.sub.a is a cycloalkyl radical as
defined herein.
[0066] "Alkylaminocarbonyl" refers to a radical of the formula
--C(O)NHR.sub.a or --C(O)NR.sub.aR.sub.a, where each R.sub.a is
independently, an alkyl radical having the indicated number of
carbon atoms as defined herein.
[0067] "Cyclolkylaminocarbonyl" refers to a radical of the formula
--C(O)NHR.sub.a, where R.sub.a is a cycloalkyl radical as defined
herein.
[0068] "Aryl" refers to a hydrocarbon ring system radical
comprising hydrogen, 6 to 18 carbon atoms and at least one aromatic
ring. Exemplary aryls are hydrocarbon ring system radical
comprising hydrogen and 6 to 9 carbon atoms and at least one
aromatic ring; hydrocarbon ring system radical comprising hydrogen
and 9 to 12 carbon atoms and at least one aromatic ring;
hydrocarbon ring system radical comprising hydrogen and 12 to 15
carbon atoms and at least one aromatic ring; or hydrocarbon ring
system radical comprising hydrogen and 15 to 18 carbon atoms and at
least one aromatic ring. For purposes of this invention, the aryl
radical may be a monocyclic, bicyclic, tricyclic or tetracyclic
ring system, which may include fused or bridged ring systems. Aryl
radicals include, but are not limited to, aryl radicals derived
from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene,
azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene,
s-indacene, indane, indene, naphthalene, phenalene, phenanthrene,
pleiadene, pyrene, and triphenylene. "Optionally substituted aryl"
refers to a aryl group or a substituted aryl group.
[0069] "Arylene" denotes divalent aryl, and "substituted arylene"
refers to divalent substituted aryl.
[0070] "Aralkyl" or "araalkylene" may be used interchangeably and
refer to a radical of the formula --R.sub.b--R.sub.c where R.sub.b
is an alkylene chain as defined herein and R.sub.c is one or more
aryl radicals as defined herein, for example, benzyl,
diphenylmethyl and the like.
[0071] "Cycloalkyl" refers to a stable non-aromatic monocyclic or
polycyclic hydrocarbon radical consisting solely of carbon and
hydrogen atoms, which may include fused or bridged ring systems,
having from three to fifteen carbon atoms, preferably having from
three to ten carbon atoms, three to nine carbon atoms, three to
eight carbon atoms, three to seven carbon atoms, three to six
carbon atoms, three to five carbon atoms, a ring with four carbon
atoms, or a ring with three carbon atoms. The cycloalkyl ring may
be saturated or unsaturated and attached to the rest of the
molecule by a single bond. Monocyclic radicals include, for
example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, and cyclooctyl. Polycyclic radicals include, for
example, adamantyl, norbornyl, decalinyl,
7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like.
[0072] "Cycloalkylalkylene" or "cycloalkylalkyl" may be used
interchangeably and refer to a radical of the formula
--R.sub.bR.sub.e where R.sub.b is an alkylene chain as defined
herein and R.sub.e is a cycloalkyl radical as defined herein. In
certain embodiments, R.sub.b is further substituted with a
cycloalkyl group, such that the cycloalkylalkylene comprises two
cycloalkyl moieties. Cyclopropylalkylene and cyclobutylalkylene are
exemplary cycloalkylalkylene groups, comprising at least one
cyclopropyl or at least one cyclobutyl group, respectively.
[0073] "Fused" refers to any ring structure described herein which
is fused to an existing ring structure in the compounds of the
invention. When the fused ring is a heterocyclyl ring or a
heteroaryl ring, any carbon atom on the existing ring structure
which becomes part of the fused heterocyclyl ring or the fused
heteroaryl ring may be replaced with a nitrogen atom.
[0074] "Halo" or "halogen" refers to bromo (bromine), chloro
(chlorine), fluoro (fluorine), or iodo (iodine).
[0075] "Haloalkyl" refers to an alkyl radical having the indicated
number of carbon atoms, as defined herein, wherein one or more
hydrogen atoms of the alkyl group are substituted with a halogen
(halo radicals), as defined above. The halogen atoms can be the
same or different. Exemplary haloalkyls are trifluoromethyl,
difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl,
1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and
the like.
[0076] "Heterocyclyl", heterocycle", or "heterocyclic ring" refers
to a stable 3- to 18-membered saturated or unsaturated radical
which consists of two to twelve carbon atoms and from one to six
heteroatoms, for example, one to five heteroatoms, one to four
heteroatoms, one to three heteroatoms, or one to two heteroatoms
selected from the group consisting of nitrogen, oxygen and sulfur.
Exemplary heterocycles include without limitation stable 3-15
membered saturated or unsaturated radicals, stable 3-12 membered
saturated or unsaturated radicals, stable 3-9 membered saturated or
unsaturated radicals, stable 8-membered saturated or unsaturated
radicals, stable 7-membered saturated or unsaturated radicals,
stable 6-membered saturated or unsaturated radicals, or stable
5-membered saturated or unsaturated radicals.
[0077] Unless stated otherwise specifically in the specification,
the heterocyclyl radical may be a monocyclic, bicyclic, tricyclic
or tetracyclic ring system, which may include fused or bridged ring
systems; and the nitrogen, carbon or sulfur atoms in the
heterocyclyl radical may be optionally oxidized; the nitrogen atom
may be optionally quaternized; and the heterocyclyl radical may be
partially or fully saturated. Examples of non-aromatic heterocyclyl
radicals include, but are not limited to, azetidinyl, dioxolanyl,
thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,
imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,
octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,
2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,
piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl,
quinuclidinyl, thiazolidinyl, tetrahydrofuryl, thietanyl,
trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl,
1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Heterocyclyls
include heteroaryls as defined herein, and examples of aromatic
heterocyclyls are listed in the definition of heteroaryls
below.
[0078] "Heterocyclylalkyl" or "heterocyclylalkylene" refers to a
radical of the formula --R.sub.bR.sub.f where R.sub.b is an
alkylene chain as defined herein and R.sub.f is a heterocyclyl
radical as defined above, and if the heterocyclyl is a
nitrogen-containing heterocyclyl, the heterocyclyl may be attached
to the alkyl radical at the nitrogen atom.
[0079] "Heteroaryl" or "heteroarylene" refers to a 5- to
14-membered ring system radical comprising hydrogen atoms, one to
thirteen carbon atoms, one to six heteroatoms selected from the
group consisting of nitrogen, oxygen and sulfur, and at least one
aromatic ring. For purposes of this invention, the heteroaryl
radical may be a stable 5-12 membered ring, a stable 5-10 membered
ring, a stable 5-9 membered ring, a stable 5-8 membered ring, a
stable 5-7 membered ring, or a stable 6 membered ring that
comprises at least 1 heteroatom, at least 2 heteroatoms, at least 3
heteroatoms, at least 4 heteroatoms, at least 5 heteroatoms or at
least 6 heteroatoms. Heteroaryls may be a monocyclic, bicyclic,
tricyclic or tetracyclic ring system, which may include fused or
bridged ring systems; and the nitrogen, carbon or sulfur atoms in
the heteroaryl radical may be optionally oxidized; the nitrogen
atom may be optionally quaternized. The heteroatom may be a member
of an aromatic or non-aromatic ring, provided at least one ring in
the heteroaryl is aromatic. Examples include, but are not limited
to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl,
benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl,
benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl,
1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl,
benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl,
benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl),
benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl,
cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl,
isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl,
isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl,
isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl,
oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl,
1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl,
phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl,
pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl,
pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl,
isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl,
triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e.
thienyl).
[0080] "Heteroarylalkyl" or "heteroarylalkylene" refers to a
radical of the formula --R.sub.bR.sub.g where R.sub.b is an
alkylene chain as defined above and R.sub.g is a heteroaryl radical
as defined above.
[0081] "Thioalkyl" refers to a radical of the formula --SR.sub.a
where R.sub.a is an alkyl radical as defined above containing one
to twelve carbon atoms, at least 1-10 carbon atoms, at least 1-8
carbon atoms, at least 1-6 carbon atoms, or at least 1-4 carbon
atoms.
[0082] "Heterocyclylaminyl" refers to a radical of the formula
--NHR.sub.f where R.sub.f is a heterocyclyl radical as defined
above.
[0083] "Thione" refers to a .dbd.S group attached to a carbon atom
of a saturated or unsaturated (C.sub.3-C.sub.8)cyclic or a
(C.sub.1-C.sub.8)acyclic moiety.
[0084] "Sulfoxide" refers to a --S(O)-- group in which the sulfur
atom is covalently attached to two carbon atoms.
[0085] "Sulfone" refers to a --S(O).sub.2-- group in which a
hexavalent sulfur is attached to each of the two oxygen atoms
through double bonds and is further attached to two carbon atoms
through single covalent bonds.
[0086] The term "oxime" refers to a --C(R.sub.a).dbd.N--OR.sub.a
radical where R.sub.a is hydrogen, lower alkyl, an alkylene or
arylene group as defined above.
[0087] The compound of the invention can exist in various isomeric
forms, as well as in one or more tautomeric forms, including both
single tautomers and mixtures of tautomers. The term "isomer" is
intended to encompass all isomeric forms of a compound of this
invention, including tautomeric forms of the compound.
[0088] Some compounds described here can have asymmetric centers
and therefore exist in different enantiomeric and diastereomeric
forms. A compound of the invention can be in the form of an optical
isomer or a diastereomer. Accordingly, the invention encompasses
compounds of the invention and their uses as described herein in
the form of their optical isomers, diastereoisomers and mixtures
thereof, including a racemic mixture. Optical isomers of the
compounds of the invention can be obtained by known techniques such
as asymmetric synthesis, chiral chromatography, or via chemical
separation of stereoisomers through the employment of optically
active resolving agents.
[0089] Unless otherwise indicated "stereoisomer" means one
stereoisomer of a compound that is substantially free of other
stereoisomers of that compound. Thus, a stereomerically pure
compound having one chiral center will be substantially free of the
opposite enantiomer of the compound. A stereomerically pure
compound having two chiral centers will be substantially free of
other diastereomers of the compound. A typical stereomerically pure
compound comprises greater than about 80% by weight of one
stereoisomer of the compound and less than about 20% by weight of
other stereoisomers of the compound, for example greater than about
90% by weight of one stereoisomer of the compound and less than
about 10% by weight of the other stereoisomers of the compound, or
greater than about 95% by weight of one stereoisomer of the
compound and less than about 5% by weight of the other
stereoisomers of the compound, or greater than about 97% by weight
of one stereoisomer of the compound and less than about 3% by
weight of the other stereoisomers of the compound.
[0090] If there is a discrepancy between a depicted structure and a
name given to that structure, then the depicted structure controls.
Additionally, if the stereochemistry of a structure or a portion of
a structure is not indicated with, for example, bold or dashed
lines, the structure or portion of the structure is to be
interpreted as encompassing all stereoisomers of it. In some cases,
however, where more than one chiral center exists, the structures
and names may be represented as single enantiomers to help describe
the relative stereochemistry. Those skilled in the art of organic
synthesis will know if the compounds are prepared as single
enantiomers from the methods used to prepare them.
[0091] In this description a "pharmaceutically acceptable salt" is
a pharmaceutically acceptable, organic or inorganic acid or base
salt of a compound of the invention. Representative
pharmaceutically acceptable salts include, e.g., alkali metal
salts, alkali earth salts, ammonium salts, water-soluble and
water-insoluble salts, such as the acetate, amsonate
(4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzonate,
bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate,
calcium, calcium edetate, camsylate, carbonate, chloride, citrate,
clavulariate, dihydrochloride, edetate, edisylate, estolate,
esylate, fiunarate, gluceptate, gluconate, glutamate,
glycollylarsanilate, hexafluorophosphate, hexylresorcinate,
hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate,
iodide, isothionate, lactate, lactobionate, laurate, malate,
maleate, mandelate, mesylate, methylbromide, methylnitrate,
methylsulfate, mucate, napsylate, nitrate, N-methylglucamine
ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate,
pamoate (1,1-methene-bis-2-hydroxy-3-naphthoate, einbonate),
pantothenate, phosphate/diphosphate, picrate, polygalacturonate,
propionate, p-toluenesulfonate, salicylate, stearate, subacetate,
succinate, sulfate, sulfosaliculate, suramate, tannate, tartrate,
teoclate, tosylate, triethiodide, and valerate salts. A
pharmaceutically acceptable salt can have more than one charged
atom in its structure. In this instance the pharmaceutically
acceptable salt can have multiple counterions. Thus, a
pharmaceutically acceptable salt can have one or more charged atoms
and/or one or more counterions.
[0092] The terms "treat", "treating" and "treatment" refer to the
amelioration or eradication of a disease or symptoms associated
with a disease. In certain embodiments, such terms refer to
minimizing the spread or worsening of the disease resulting from
the administration of one or more prophylactic or therapeutic
agents to a patient with such a disease. In the context of the
present invention the terms "treat", "treating" and "treatment"
also refer to:
(i) preventing the disease or condition from occurring in a mammal,
in particular, when such mammal is predisposed to the condition but
has not yet been diagnosed as having it; (ii) inhibiting the
disease or condition, i.e., arresting its development; (iii)
relieving the disease or condition, i.e., causing regression of the
disease or condition; or (iv) relieving the symptoms resulting from
the disease or condition, i.e., relieving pain without addressing
the underlying disease or condition. As used herein, the terms
"disease" and "condition" may be used interchangeably or may be
different in that the particular malady or condition may not have a
known causative agent (so that etiology has not yet been worked
out) and it is therefore not yet recognized as a disease but only
as an undesirable condition or syndrome, wherein a more or less
specific set of symptoms have been identified by clinicians.
[0093] The term "effective amount" refers to an amount of a
compound of the invention or other active ingredient sufficient to
provide a therapeutic or prophylactic benefit in the treatment or
prevention of a disease or to delay or minimize symptoms associated
with a disease. Further, a therapeutically effective amount with
respect to a compound of the invention means that amount of
therapeutic agent alone, or in combination with other therapies,
that provides a therapeutic benefit in the treatment or prevention
of a disease. Used in connection with a compound of the invention,
the term can encompass an amount that improves overall therapy,
reduces or avoids symptoms or causes of disease, or enhances the
therapeutic efficacy or synergies with another therapeutic
agent.
[0094] The terms "modulate", "modulation" and the like refer to the
ability of a compound to increase or decrease the function, or
activity of, for example, MAP kinase interacting kinase (Mnk).
"Modulation", in its various forms, is intended to encompass
inhibition, antagonism, partial antagonism, activation, agonism
and/or partial agonism of the activity associated with Mnk. Mnk
inhibitors are compounds that bind to, partially or totally block
stimulation, decrease, prevent, delay activation, inactivate,
desensitize, or down regulate signal transduction. The ability of a
compound to modulate Mnk activity can be demonstrated in an
enzymatic assay or a cell-based assay.
[0095] A "patient" or subject" includes an animal, such as a human,
cow, horse, sheep, lamb, pig, chicken, turkey, quail, cat, dog,
mouse, rat, rabbit or guinea pig. The animal can be a mammal such
as a non-primate and a primate (e.g., monkey and human). In one
embodiment, a patient is a human, such as a human infant, child,
adolescent or adult.
[0096] The term "prodrug" refers to a precursor of a drug, a
compound which upon administration to a patient, must undergo
chemical conversion by metabolic processes before becoming an
active pharmacological agent. Exemplary prodrugs of compounds in
accordance with Formula I are esters, acetamides, and amides.
Compounds of the Invention
[0097] The present invention generally is directed to compounds
encompassed by the genus of Formula I, a stereoisomer, a tautomer
or a pharmaceutically acceptable salt thereof.
##STR00003##
[0098] For Formula I compounds A.sup.1, A.sup.2, A.sup.3, A.sup.4,
A.sup.5, A.sup.6, A.sup.7, W.sup.1, R.sup.1, R.sup.2, R.sup.3,
R.sup.4, R.sup.5a, R.sup.5b, R.sup.6, R.sup.7, R.sup.7a, R.sup.7b,
R.sup.8, R.sup.8a, R.sup.8b, R.sup.9, R.sup.9a, R.sup.9b and
R.sup.10 and subscripts "m" and "n" are as defined in the
specification. Described below are specific embodiments of Formula
I compounds.
[0099] In one embodiment A.sup.1 and A.sup.2 are --CR.sup.5a.
[0100] In another embodiment A.sup.1 is --N-- and A.sup.2 is --CH
or --C(Me). In yet another embodiment A.sup.1 is --CH and A.sup.2
is --N--.
[0101] In one embodiment A.sup.3 is --CR.sup.6.
[0102] In one embodiment A.sup.4 is --CR.sup.5b.
[0103] In one embodiment A.sup.5 is an --NR.sup.7.
[0104] In another embodiment A.sup.5 is --CR.sup.7aR.sup.7b.
[0105] In one embodiment A.sup.6 and A.sup.7 are --CR.sup.8a.
[0106] In one embodiment W.sup.1 is O.
[0107] In one embodiment subscript "m" and subscript "n" are 1.
[0108] In another embodiment subscript "m" is 2 and subscript "n"
is 1. In yet another embodiment subscripts "m" and "n" are both
2.
[0109] In one embodiment R.sup.1 and R.sup.2 independently are --H,
(C.sub.1-C.sub.8)alkyl, --NHR.sup.10 or NHR.sup.10-alkylene.
[0110] In one embodiment R.sup.1 and R.sup.2 are --H, --NH.sub.2,
--NH(Me), --N(Me).sub.2, methyl, ethyl, propyl, isopropyl, butyl,
sec-butyl, t-butyl, isobutyl, pentyl, hexyl, methylene-NH[C(O)OMe]
or ethylene-NH[C(O)OMe].
[0111] In one embodiment at least one of R.sup.1 or R.sup.2 is a
halogen substituted (C.sub.1-C.sub.8)alkyl,
(C.sub.2-C.sub.8)alkenyl, (C.sub.2-C.sub.8)alkynyl, cycloalkyl,
heterocyclyl, heteroaryl, aryl, arylalkylene, cycloalkylalkylene,
heterocyclylalkylene or heteroarylalkylene.
[0112] In one embodiment R.sup.1 and R.sup.2 together with the
carbon atom to which they are attached form a cycloalkyl ring. In
an embodiment the cycloalkyl ring is cyclobutyl, cyclopentyl,
cyclohexyl, 2,2-dimethylcyclobutyl, 4-aminocyclohexyl,
4-methylcyclohexyl, 4-ethylcyclohexyl,
2,2-difluoroethyl-4-cyclohexyl, 4,4-difluorocyclohexy,
4-cyanocyclohexyl, 4-trifluoromethylcyclohexyl,
4-hydroxycyclohexyl, 3-hydroxycyclopently, 3-aminocyclopentyl or a
3-methylcyclopentyl ring system. In yet another embodiment the
cycloalkyl ring is cyclobutyl, cyclopentyl or cyclohexyl.
[0113] In one embodiment R.sup.1 and R.sup.2 together with the
carbon atom to which they are attached form a heterocyclyl ring. In
an embodiment the heterocyclyl ring is
1-(2,2-difluoroethyl)piperidine or 1-methylpiperidine.
[0114] In one embodiment R.sup.3, R.sup.4, and R.sup.5b
independently are --O(C.sub.1-C.sub.8)alkyleneNHR.sup.10 or
--O(C.sub.1-C.sub.8)alkyleneNR.sup.10R.sup.10. In another
embodiment R.sup.3, R.sup.4, and R.sup.5b independently are
--O(CH.sub.2)NH.sub.2, --O(CH.sub.2).sub.2NH.sub.2,
--O(CH.sub.2).sub.3NH.sub.2, --O(CH.sub.2)NH(Me),
--O(CH.sub.2).sub.2NH(Me) or --O(CH.sub.2).sub.3NH(Me). In yet
another embodiment R.sup.3, R.sup.4, and R.sup.Sb independently are
--O(CH.sub.2)N(Me).sub.2, --O(CH.sub.2).sub.2N(Me).sub.2 or
--O(CH.sub.2).sub.3N(Me).sub.2.
[0115] In one embodiment R.sup.3, R.sup.4, and R.sup.5b
independently are --H, --OH, CN, --C(O)NH.sub.2, --C(O)NH(Me),
--NH.sub.2, --NH(Me), --N(Me).sub.2, --NH.sub.2-methylene,
--NH.sub.2-ethylene, methyl, ethyl, propyl, n-butyl, i-butyl,
t-butyl, hexyl, methoxy, ethoxy, propoxy, butoxy, chloromethyl,
fluoromethyl, dichloromethyl, chlorofluoromethyl, trifluoromethyl,
chloroethyl, 1,2-dichloroethyl or chlorofluoroethyl.
[0116] In one embodiment R.sup.5a is --H, --OH, halogen, --CN,
acetyl or --(C.sub.1-C.sub.8)alkyl. In another embodiment R.sup.5a
is methyl, ethyl, propyl or butyl.
[0117] In one embodiment R.sup.6 is amino, methylamino, --CN,
--O(C.sub.1-C.sub.8)alkyleneNHR.sup.10,
--O(C.sub.1-C.sub.8)alkyleneNR.sup.10R.sup.10,
--(C.sub.1-C.sub.8)alkyleneNHR.sup.10 or
--(C.sub.1-C.sub.8)alkyleneNR.sup.10R.sup.10.
[0118] In another embodiment R.sup.6 is --H, --OH, chlorine,
fluorine, methyl ethyl, propyl and the like.
[0119] In one embodiment R.sup.7, R.sup.7a and R.sup.7b are
hydrogen.
[0120] In one embodiment R.sup.8, R.sup.8a and R.sup.8b
independently are --H, heterocyclyl, heteroaryl or aryl. In another
embodiment R.sup.8, R.sup.8a and R.sup.8b independently are
pyridine, 1-(2, 2-difluoroethyl)piperidine, 1-difluoromethyl
piperidine, N-methylpyrazole, thioimidazole, piperidine or
N-methylpiperidine, phenyl, 2-chlorophenyl, 3-chlorophenyl,
4-chlorophenyl, 2-cyanophenyl, 3-cyanophenyl or 4-cyanophenyl.
[0121] In one embodiment R.sup.8a and R.sup.8b independently are
--H, --OH, --CN, Cl, F, methyl, ethyl propyl, chloromethyl,
fluoromethyl, chlorofluoromethyl, --NH(Me) or --N(Me).sub.2.
[0122] In one embodiment R.sup.9, R.sup.9a and R.sup.9b are
independently --H or --(C.sub.1-C.sub.8)alkyl.
[0123] In one embodiment R.sup.10 is --H, --OH, methyl, ethyl,
propyl, butyl, t-butyl, acetyl, --COOMe, --NH.sub.2, --NH(Me), or
--N(Me).sub.2.
[0124] In one embodiment A.sup.1 is --N, A.sup.2, A.sup.3, A.sup.4,
A.sup.6, and A.sup.7 are --CH, A.sup.5 is --NH, W.sup.1 is O, and
subscripts "m" and "n" are both 1.
[0125] In another embodiment A.sup.1 is --N, A.sup.2 is --CH,
A.sup.3 is --C(Cl), --C(F), --C(Me) or --C(OH), A.sup.4, A.sup.6,
and A.sup.7 are --CH, A.sup.5 is --NH, W.sup.1 is O, and subscripts
"m" and "n" are both 1.
[0126] In another embodiment A.sup.1 is --N, A.sup.2 is --CH,
A.sup.3 is --C(alkyl) or --C(halogen), A.sup.4 is --CH, A.sup.5 is
--NH, A.sup.6 and A.sup.7 are both --CR.sup.8a, W.sup.1 is O, and
subscripts "m" and "n" are both 1. In an embodiment --CR.sup.8a is
--C(pyridyl), --C(N-methylpyrazole), --C(2-chlorophenyl) or
--C(2-cyanophenyl).
[0127] In another embodiment A.sup.1 is --N, A.sup.2, A.sup.3,
A.sup.4 are --CH, A.sup.5 is --NH, A.sup.6 and A.sup.7 are --N--,
W.sup.1 is O, and subscripts "m" and "n" are both 1.
[0128] In another embodiment one of A.sup.6 or A.sup.7 is --N and
the other of A.sup.6 or A.sup.7 is --CH, --C(pyridyl),
--C(N-methylpyrazole), --C(2-chlorophenyl) or
--C(2-cyanophenyl).
[0129] In another embodiment A.sup.1 is --N, A.sup.2, A.sup.3,
A.sup.4 are --CH, A.sup.5 is --NH, A.sup.6 and A.sup.7
independently are CH.sub.2-- or --CH(Me), W.sup.1 is O, and
subscripts "m" and "n" are both 1.
[0130] In another embodiment one of A.sup.6 or A.sup.7 is
--CH.sub.2 or --CH(Me) and the other of A.sup.6 or A.sup.7 is
--NH.
[0131] In one embodiment subscript "m" is 2 and subscript "n" is 1,
A.sup.1 is --N, A.sup.2, A.sup.3 and A.sup.4 are --CH, A.sup.5 is
--NH and A.sup.6 and A.sup.7 are --CH.sub.2.
[0132] The inventive compounds according to Formula I may be
isotopically-labeled by having one or more atoms replaced by an
atom having a different atomic mass or mass number. Examples of
isotopes that can be incorporated into compounds of according to
Formula I include isotopes of hydrogen, carbon, nitrogen, oxygen,
phosphorous, fluorine, chlorine, or iodine. Illustrative of such
isotopes are .sup.2H, .sup.3H, .sup.11C, .sup.13C, .sup.14C,
.sup.13N, .sup.15N, .sup.15O, .sup.17O, .sup.18O, .sup.31P,
.sup.32P, .sup.35S, .sup.18F, .sup.36Cl, .sup.123I, and .sup.125I,
respectively. These radiolabeled compounds can be used to measure
the biodistribution, tissue concentration and the kinetics of
transport and excretion from biological tissues including a subject
to which such a labeled compound is administered. Labeled compounds
are also used to determine therapeutic effectiveness, the site or
mode of action, and the binding affinity of a candidate therapeutic
to a pharmacologically important target. Certain
radioactive-labeled compounds according to Formula I, therefore,
are useful in drug and/or tissue distribution studies. The
radioactive isotopes tritium, i.e. .sup.3H, and carbon-14, i.e.
.sup.14C, are particularly useful for this purpose in view of their
ease of incorporation and ready means of detection.
[0133] Substitution with heavier isotopes such as deuterium, i.e.
.sup.2H, affords certain therapeutic advantages resulting from the
greater metabolic stability, for example, increased in vivo
half-life of compounds containing deuterium. Substitution of
hydrogen with deuterium may reduce dose required for therapeutic
effect, and hence may be preferred in a discovery or clinical
setting.
[0134] Substitution with positron emitting isotopes, such as
.sup.11C, .sup.18F, .sup.15O and .sup.13N, provides labeled analogs
of the inventive compounds that are useful in Positron Emission
Tomography (PET) studies, e.g., for examining substrate receptor
occupancy. Isotopically-labeled compounds according to Formula I,
can generally be prepared by conventional techniques known to those
skilled in the art or by processes analogous to those described in
the Preparations and Examples section as set out below using an
appropriate isotopic-labeling reagent.
[0135] Embodiments of the invention disclosed herein are also meant
to encompass the in vivo metabolic products of compounds according
to Formula I. Such products may result from, for example, the
oxidation, reduction, hydrolysis, amidation, esterification, and
like processes primarily due to enzymatic activity upon
administration of a compound of the invention. Accordingly, the
invention includes compounds that are produced as by-products of
enzymatic or non-enzymatic activity on an inventive compound
following the administration of such a compound to a mammal for a
period of time sufficient to yield a metabolic product. Metabolic
products, particularly pharmaceutically active metabolites are
typically identified by administering a radiolabeled compound of
the invention in a detectable dose to a subject, such as rat,
mouse, guinea pig, monkey, or human, for a sufficient period of
time during which metabolism occurs, and isolating the metabolic
products from urine, blood or other biological samples that are
obtained from the subject receiving the radiolabeled compound.
[0136] The invention also provides pharmaceutically acceptable salt
forms of Formula I compounds. Encompassed within the scope of the
invention are both acid and base addition salts that are formed by
contacting a pharmaceutically suitable acid or a pharmaceutically
suitable base with a compound of the invention.
[0137] To this end, a "pharmaceutically acceptable acid addition
salt" refers to those salts which retain the biological
effectiveness and properties of the free bases, which are not
biologically or otherwise undesirable, and which are formed with
inorganic acids such as, but are not limited to, hydrochloric acid,
hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and
the like, and organic acids such as, but not limited to, acetic
acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic
acid, aspartic acid, benzenesulfonic acid, benzoic acid,
4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid,
capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic
acid, citric acid, cyclamic acid, dodecylsulfuric acid,
ethane-1,2-disulfonic acid, ethanesulfonic acid,
2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric
acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic
acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid,
glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric
acid, lactic acid, lactobionic acid, lauric acid, maleic acid,
malic acid, malonic acid, mandelic acid, methanesulfonic acid,
mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic
acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid,
orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic
acid, pyroglutamic acid, pyruvic acid, salicylic acid,
4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid,
tartaric acid, thiocyanic acid, p-toluenesulfonic acid,
trifluoroacetic acid, undecylenic acid, and the like.
[0138] Similarly, a "pharmaceutically acceptable base addition
salt" refers to those salts which retain the biological
effectiveness and properties of the free acids, which are not
biologically or otherwise undesirable. These salts are prepared by
addition of an inorganic base or an organic base to the free acid.
Salts derived from inorganic bases include, but are not limited to,
the sodium, potassium, lithium, ammonium, calcium, magnesium, iron,
zinc, copper, manganese, aluminum salts and the like. Preferred
inorganic salts are the ammonium, sodium, potassium, calcium, and
magnesium salts. Salts derived from organic bases include, but are
not limited to, salts of primary, secondary, and tertiary amines,
substituted amines including naturally occurring substituted
amines, cyclic amines and basic ion exchange resins, such as
ammonia, isopropylamine, trimethylamine, diethylamine,
triethylamine, tripropylamine, diethanolamine, ethanolamine,
deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol,
dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,
hydrabamine, choline, betaine, benethamine, benzathine,
ethylenediamine, glucosamine, methylglucamine, theobromine,
triethanolamine, tromethamine, purines, piperazine, piperidine,
N-ethylpiperidine, polyamine resins and the like. Particularly
preferred organic bases are isopropylamine, diethylamine,
ethanolamine, trimethylamine, dicyclohexylamine, choline and
caffeine.
[0139] Often crystallizations produce a solvate of the compound of
the invention. As used herein, the term "solvate" refers to an
aggregate that comprises one or more molecules of a compound of the
invention with one or more molecules of solvent. The solvent may be
water, in which case the solvate may be a hydrate. Alternatively,
the solvent may be an organic solvent. Thus, the compounds of the
present invention may exist as a hydrate, including a monohydrate,
dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and
the like, as well as the corresponding solvated forms. The compound
of the invention may be true solvates, while in other cases, the
compound of the invention may merely retain adventitious water or
be a mixture of water plus some adventitious solvent.
[0140] A "stereoisomer" refers to a compound made up of the same
atoms bonded by the same bonds but having different
three-dimensional structures, which are not interchangeable. The
present invention contemplates various stereoisomers and mixtures
thereof and includes "enantiomers", which refers to two
stereoisomers whose molecules are nonsuperimposeable mirror images
of one another.
[0141] Compounds of the invention or their pharmaceutically
acceptable salts may contain one or more asymmetric centers and may
thus give rise to enantiomers, diastereomers, and other
stereoisomeric forms that may be defined, in terms of absolute
stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino
acids. The present invention is meant to include all such possible
isomers, as well as their racemic and optically pure forms.
Optically active (+) and (-), (R)- and (S)-, or (D)- and
(L)-isomers may be prepared using chiral synthons or chiral
reagents, or resolved using conventional techniques, for example,
chromatography and fractional crystallization. Conventional
techniques for the preparation/isolation of individual enantiomers
include chiral synthesis from a suitable optically pure precursor
or resolution of the racemate (or the racemate of a salt or
derivative) using, for example, chiral high pressure liquid
chromatography (HPLC). When the compounds described herein contain
olefinic double bonds or other centers of geometric asymmetry, and
unless specified otherwise, it is intended that the compounds
include both E and Z geometric isomers. Likewise, all tautomeric
forms are also intended to be included.
[0142] The term "tautomer" refers to a proton shift from one atom
of a molecule to another atom of the same molecule. For example,
when W.sup.1 is oxo and A5 is --NH, the present invention provides
tautomers of a Formula I compound as illustrated below:
##STR00004##
The inventive compounds are synthesized using conventional
synthetic methods, and more specifically using the general methods
noted below. Specific synthetic protocols for compounds in
accordance with the present invention are described in the
Examples.
Pharmaceutical Formulations
[0143] In one embodiment, a compounds according Formulae I are
formulated as pharmaceutically acceptable compositions that contain
a Formulae I compound in an amount effective to treat a particular
disease or condition of interest upon administration of the
pharmaceutical composition to a mammal. Pharmaceutical compositions
in accordance with the present invention can comprise a Formulae I
compound in combination with a pharmaceutically acceptable carrier,
diluent or excipient.
[0144] In this regard, a "pharmaceutically acceptable carrier,
diluent or excipient" includes without limitation any adjuvant,
carrier, excipient, glidant, sweetening agent, diluent,
preservative, dye/colorant, flavor enhancer, surfactant, wetting
agent, dispersing agent, suspending agent, stabilizer, isotonic
agent, solvent, or emulsifier which has been approved by the United
States Food and Drug Administration as being acceptable for use in
humans or domestic animals.
[0145] Further, a "mammal" includes humans and both domestic
animals such as laboratory animals and household pets (e.g., cats,
dogs, swine, cattle, sheep, goats, horses, rabbits), and
non-domestic animals such as wildlife and the like.
[0146] The pharmaceutical compositions of the invention can be
prepared by combining a compound of the invention with an
appropriate pharmaceutically acceptable carrier, diluent or
excipient, and may be formulated into preparations in solid,
semi-solid, liquid or gaseous forms, such as tablets, capsules,
powders, granules, ointments, solutions, suppositories, injections,
inhalants, gels, microspheres, and aerosols. Typical routes of
administering such pharmaceutical compositions include, without
limitation, oral, topical, transdermal, inhalation, parenteral,
sublingual, buccal, rectal, vaginal, and intranasal. The term
parenteral as used herein includes subcutaneous injections,
intravenous, intramuscular, intrasternal injection or infusion
techniques. Pharmaceutical compositions of the invention are
formulated so as to allow the active ingredients contained therein
to be bioavailable upon administration of the composition to a
patient. Compositions that will be administered to a subject or
patient take the form of one or more dosage units, where for
example, a tablet may be a single dosage unit, and a container of a
compound of the invention in aerosol form may hold a plurality of
dosage units. Actual methods of preparing such dosage forms are
known, or will be apparent, to those skilled in this art; for
example, see Remington: The Science and Practice of Pharmacy, 20th
Edition (Philadelphia College of Pharmacy and Science, 2000). The
composition to be administered will, in any event, contain a
therapeutically effective amount of a compound of the invention, or
a pharmaceutically acceptable salt thereof, for treatment of a
disease or condition of interest in accordance with the teachings
of this invention.
[0147] A pharmaceutical composition of the invention may be in the
form of a solid or liquid. In one aspect, the carrier(s) are
particulate, so that the compositions are, for example, in tablet
or powder form. The carrier(s) may be liquid, with the compositions
being, for example, an oral syrup, injectable liquid or an aerosol,
which is useful in, for example, inhalatory administration. When
intended for oral administration, the pharmaceutical composition is
preferably in either solid or liquid form, where semi-solid,
semi-liquid, suspension and gel forms are included within the forms
considered herein as either solid or liquid.
[0148] As a solid composition for oral administration the
pharmaceutical composition may be formulated into a powder,
granule, compressed tablet, pill, capsule, chewing gum, wafer or
the like form. Such a solid composition will typically contain one
or more inert diluents or edible carriers. In addition, one or more
of the following may be present: binders such as
carboxymethylcellulose, ethyl cellulose, microcrystalline
cellulose, gum tragacanth or gelatin; excipients such as starch,
lactose or dextrins, disintegrating agents such as alginic acid,
sodium alginate, Primogel, corn starch and the like; lubricants
such as magnesium stearate or Sterotex; glidants such as colloidal
silicon dioxide; sweetening agents such as sucrose or saccharin; a
flavoring agent such as peppermint, methyl salicylate or orange
flavoring; and a coloring agent.
[0149] When the pharmaceutical composition is in the form of a
capsule, for example, a gelatin capsule, it may contain, in
addition to materials of the above type, a liquid carrier such as
polyethylene glycol or oil.
[0150] The pharmaceutical composition may be in the form of a
liquid, for example, an elixir, syrup, solution, emulsion or
suspension. The liquid may be for oral administration or for
delivery by injection, as two examples. When intended for oral
administration, preferred composition contain, in addition to the
present compounds, one or more of a sweetening agent,
preservatives, dye/colorant and flavor enhancer. In a composition
intended to be administered by injection, one or more of a
surfactant, preservative, wetting agent, dispersing agent,
suspending agent, buffer, stabilizer and isotonic agent may be
included.
[0151] The liquid pharmaceutical compositions of the invention,
whether they be solutions, suspensions or other like form, may
include one or more of the following adjuvants: sterile diluents
such as water for injection, saline solution, preferably
physiological saline, Ringer's solution, isotonic sodium chloride,
fixed oils such as synthetic mono or diglycerides which may serve
as the solvent or suspending medium, polyethylene glycols,
glycerin, propylene glycol or other solvents; antibacterial agents
such as benzyl alcohol or methyl paraben; antioxidants such as
ascorbic acid or sodium bisulfite; chelating agents such as
ethylenediaminetetraacetic acid; buffers such as acetates, citrates
or phosphates and agents for the adjustment of tonicity such as
sodium chloride or dextrose. The parenteral preparation can be
enclosed in ampoules, disposable syringes or multiple dose vials
made of glass or plastic. Physiological saline is a preferred
adjuvant. An injectable pharmaceutical composition is preferably
sterile.
[0152] A liquid pharmaceutical composition of the invention
intended for either parenteral or oral administration should
contain an amount of a compound of the invention such that a
suitable dosage will be obtained.
[0153] The pharmaceutical composition of the invention may be
intended for topical administration, in which case the carrier may
suitably comprise a solution, emulsion, ointment or gel base. The
base, for example, may comprise one or more of the following:
petrolatum, lanolin, polyethylene glycols, bee wax, mineral oil,
diluents such as water and alcohol, and emulsifiers and
stabilizers. Thickening agents may be present in a pharmaceutical
composition for topical administration. If intended for transdermal
administration, the composition may include a transdermal patch or
iontophoresis device.
[0154] The pharmaceutical composition of the invention may be
intended for rectal administration, in the form, for example, of a
suppository, which will melt in the rectum and release the drug.
The composition for rectal administration may contain an oleaginous
base as a suitable nonirritating excipient. Such bases include,
without limitation, lanolin, cocoa butter and polyethylene
glycol.
[0155] The pharmaceutical composition of the invention may include
various materials, which modify the physical form of a solid or
liquid dosage unit. For example, the composition may include
materials that form a coating shell around the active ingredients.
The materials that form the coating shell are typically inert, and
may be selected from, for example, sugar, shellac, and other
enteric coating agents. Alternatively, the active ingredients may
be encased in a gelatin capsule.
[0156] The pharmaceutical composition of the invention in solid or
liquid form may include an agent that binds to the compound of the
invention and thereby assists in the delivery of the compound.
Suitable agents that may act in this capacity include a monoclonal
or polyclonal antibody, a protein or a liposome.
[0157] The pharmaceutical composition of the invention may consist
of dosage units that can be administered as an aerosol. The term
aerosol is used to denote a variety of systems ranging from those
of colloidal nature to systems consisting of pressurized packages.
Delivery may be by a liquefied or compressed gas or by a suitable
pump system that dispenses the active ingredients. Aerosols of
compounds of the invention may be delivered in single phase,
bi-phasic, or tri-phasic systems in order to deliver the active
ingredient(s). Delivery of the aerosol includes the necessary
container, activators, valves, subcontainers, and the like, which
together may form a kit. One skilled in the art, without undue
experimentation may determine preferred aerosols.
[0158] The pharmaceutical compositions of the invention may be
prepared by any methodology well known in the pharmaceutical art.
For example, a pharmaceutical composition intended to be
administered by injection can be prepared by combining a compound
of the invention with sterile, distilled water so as to form a
solution. A surfactant may be added to facilitate the formation of
a homogeneous solution or suspension. Surfactants are compounds
that non-covalently interact with the compound of the invention so
as to facilitate dissolution or homogeneous suspension of the
compound in the aqueous delivery system.
[0159] In certain embodiments a pharmaceutical composition
comprising a compound of Formula I is administered to a mammal in
an amount sufficient to inhibit Mnk activity upon administration,
and preferably with acceptable toxicity to the same. Mnk activity
of Formula I compounds can be determined by one skilled in the art,
for example, as described in the Examples below. Appropriate
concentrations and dosages can be readily determined by one skilled
in the art.
Therapeutic Use
[0160] The compounds of the invention, or their pharmaceutically
acceptable salts, are administered in a therapeutically effective
amount, which will vary depending upon a variety of factors
including the activity of the specific compound employed; the
metabolic stability and length of action of the compound; the age,
body weight, general health, sex, and diet of the patient; the mode
and time of administration; the rate of excretion; the drug
combination; the severity of the particular disorder or condition;
and the subject undergoing therapy.
[0161] "Effective amount" or "therapeutically effective amount"
refers to that amount of a compound of the invention which, when
administered to a mammal, preferably a human, is sufficient to
effect treatment, as defined below, of a Mnk related condition or
disease in the mammal, preferably a human. The amount of a compound
of the invention which constitutes a "therapeutically effective
amount" will vary depending on the compound, the condition and its
severity, the manner of administration, and the age of the mammal
to be treated, but can be determined routinely by one of ordinary
skill in the art having regard to his own knowledge and to this
disclosure.
[0162] Compounds of the invention or pharmaceutically acceptable
salt thereof may also be administered simultaneously with, prior
to, or after administration of one or more other therapeutic
agents. Such combination therapy includes administration of a
single pharmaceutical dosage formulation which contains a compound
of the invention and one or more additional active agents, as well
as administration of the compound of the invention and each active
agent in its own separate pharmaceutical dosage formulation. For
example, a compound of the invention and the other active agent can
be administered to the patient together in a single oral dosage
composition such as a tablet or capsule, or each agent administered
in separate oral dosage formulations. Where separate dosage
formulations are used, the compounds of the invention and one or
more additional active agents can be administered at essentially
the same time, i.e., concurrently, or at separately staggered
times, i.e., sequentially; combination therapy is understood to
include all these regimens.
[0163] In certain embodiments the disclosed compounds are useful
for inhibiting the activity of Mnk and/or can be useful in
analyzing Mnk signaling activity in model systems and/or for
preventing, treating, or ameliorating a symptom associated with a
disease, disorder, or pathological condition involving Mnk,
preferably one afflicting humans. A compound which inhibits the
activity of Mnk will be useful in preventing, treating,
ameliorating, or reducing the symptoms or progression of diseases
of uncontrolled cell growth, proliferation and/or survival,
inappropriate cellular immune responses, or inappropriate cellular
inflammatory responses or diseases which are accompanied with
uncontrolled cell growth, proliferation and/or survival,
inappropriate cellular immune responses, or inappropriate cellular
inflammatory responses, particularly in which the uncontrolled cell
growth, proliferation and/or survival, inappropriate cellular
immune responses, or inappropriate cellular inflammatory responses
is mediated by Mnk, such as, for example, haematological tumors,
solid tumors, and/or metastases thereof, including leukaemias and
myelodysplastic syndrome, Waldenstrom macroglobulinemia, and
malignant lymphomas, for example, B-cell lymphoma, T-cell lymphoma,
hairy cell lymphoma, Hodgkins lymphoma, non-Hodgins lymphoma, and
Burketts lymphoma, head and neck tumors including brain tumors and
brain metastases, tumors of the thorax including non-small cell and
small cell lung tumors, gastrointestinal tumors, endocrine tumors,
mammary and other gynecological tumors, urological tumors including
renal, bladder and prostate tumors, skin tumors, and sarcomas,
and/or metastases thereof.
[0164] Furthermore, the inventive compounds and their
pharmaceutical compositions are candidate therapeutics for the
prophylaxis and/or therapy of cytokine related diseases, such as
inflammatory diseases, allergies, or other conditions associated
with proinflammatory cytokines. Exemplary inflammatory diseases
include without limitation, chronic or acute inflammation,
inflammation of the joints such as chronic inflammatory arthritis,
rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile
rheumatoid arthritis, Reiter's syndrome, rheumatoid traumatic
arthritis, rubella arthritis, acute synovitis and gouty arthritis;
inflammatory skin diseases such as sunburn, psoriasis,
erythrodermic psoriasis, pustular psoriasis, eczema, dermatitis,
acute or chronic graft formation, atopic dermatitis, contact
dermatitis, urticaria and scleroderma; inflammation of the
gastrointestinal tract such as inflammatory bowel disease, Crohn's
disease and related conditions, ulcerative colitis, colitis, and
diverticulitis; nephritis, urethritis, salpingitis, oophoritis,
endomyometritis, spondylitis, systemic lupus erythematosus and
related disorders, multiple sclerosis, asthma, meningitis,
myelitis, encephalomyelitis, encephalitis, phlebitis,
thrombophlebitis, respiratory diseases such as asthma, bronchitis,
chronic obstructive pulmonary disease (COPD), inflammatory lung
disease and adult respiratory distress syndrome, and allergic
rhinitis; endocarditis, osteomyelitis, rheumatic fever, rheumatic
pericarditis, rheumatic endocarditis, rheumatic myocarditis,
rheumatic mitral valve disease, rheumatic aortic valve disease,
prostatitis, prostatocystitis, spondoarthropathies ankylosing
spondylitis, synovitis, tenosynovotis, myositis, pharyngitis,
polymyalgia rheumatica, shoulder tendonitis or bursitis, gout,
pseudo gout, vasculitides, inflammatory diseases of the thyroid
selected from granulomatous thyroiditis, lymphocytic thyroiditis,
invasive fibrous thyroiditis, acute thyroiditis; Hashimoto's
thyroiditis, Kawasaki's disease, Raynaud's phenomenon, Sjogren's
syndrome, neuroinflammatory disease, sepsis, conjunctivitis,
keratitis, iridocyclitis, optic neuritis, otitis, lymphoadenitis,
nasopaharingitis, sinusitis, pharyngitis, tonsillitis, laryngitis,
epiglottitis, bronchitis, pneumonitis, stomatitis, gingivitis.
oesophagitis, gastritis, peritonitis, hepatitis, cholelithiasis,
cholecystitis, glomerulonephritis, goodpasture's disease,
crescentic glomerulonephritis, pancreatitis, endomyometritis,
myometritis, metritis, cervicitis, endocervicitis, exocervicitis,
parametritis, tuberculosis, vaginitis, vulvitis, silicosis,
sarcoidosis, pneumoconiosis, pyresis, inflammatory
polyarthropathies, psoriatric arthropathies, intestinal fibrosis,
bronchiectasis and enteropathic arthropathies.
[0165] Although inflammation is the unifying pathogenic process of
these diseases, current therapies only treat the symptoms of the
disease and not the underlying cause of inflammation. The
compositions of the present invention are useful for the treatment
and/or prophylaxis of inflammatory diseases and related
complications and disorders.
[0166] Accordingly, certain embodiments are directed to a method
for treating a Mnk dependent condition in a mammal in need thereof,
the method comprising administering an effective amount of a
pharmaceutical composition as described above (i.e., a
pharmaceutical composition comprising any one or more compounds of
Formula I) to a mammal.
[0167] As described above deregulation of protein synthesis is a
common event in human cancers. A key regulator of translational
control is eIF4E whose activity is a key determinant of
tumorigenicity. Because activation of eIF4E involves
phosphorylation of a key serine (Ser209) specifically by MAP kinase
interacting kinases (Mnk), inhibitors of Mnk are suitable candidate
therapeutics for treating cell proliferative disorders such as
cancer. A wide variety of cancers, including solid tumors,
lymphomas and leukemias, are amenable to the compositions and
methods disclosed herein. Types of cancer that may be treated
include, but are not limited to: adenocarcinoma of the breast,
prostate and colon; all forms of bronchogenic carcinoma of the
lung; myeloid; melanoma; hepatoma; neuroblastoma; papilloma;
apudoma; choristoma; branchioma; malignant carcinoid syndrome;
carcinoid heart disease; and carcinoma (e.g., Walker, basal cell,
basosquamous, Brown-Pearce, ductal, Ehrlich tumor, Krebs 2, merkel
cell, mucinous, non-small cell lung, oat cell, papillary,
scirrhous, bronchiolar, bronchogenic, squamous cell, and
transitional cell). Additional types of cancers that may be treated
include: histiocytic disorders; acute and chronic leukemia, both
myeloid and lymphoid/lymphoblastic, including hairy cell leukemia;
histiocytosis malignant; Hodgkins disease; immunoproliferative
small; Hodgkins lymphoma; B-cell and T-cell non-Hodgkins lymphoma,
including diffuse large B-cell and Burkett's lymphoma;
plasmacytoma; reticuloendotheliosis; melanoma; multiple myeloma;
chondroblastoma; chondroma; chondrosarcoma; fibroma; fibrosarcoma;
myelofibrosis; giant cell tumors; histiocytoma; lipoma;
liposarcoma; mesothelioma; myxoma; myxosarcoma; osteoma;
osteosarcoma; chordoma; craniopharyngioma; dysgerminoma; hamartoma;
mesenchymoma; mesonephroma; myosarcoma; ameloblastoma; cementoma;
odontoma; teratoma; thymoma; trophoblastic tumor.
[0168] Other cancers that can be treated using the inventive
compounds include without limitation adenoma; cholangioma;
cholesteatoma; cyclindroma; cystadenocarcinoma; cystadenoma;
granulosa cell tumor; gynandroblastoma; hepatoma; hidradenoma;
islet cell tumor; Leydig cell tumor; papilloma; sertoli cell tumor;
theca cell tumor; leimyoma; leiomyosarcoma; myoblastoma; myomma;
myosarcoma; rhabdomyoma; rhabdomyosarcoma; ependymoma;
ganglioneuroma; glioma; medulloblastoma; meningioma; neurilemmoma;
neuroblastoma; neuroepithelioma; neurofibroma; neuroma;
paraganglioma; paraganglioma nonchromaffin.
[0169] In one embodiment the inventive compounds are candidate
therapeutic agents for the treatment of cancers such as
angiokeratoma; angiolymphoid hyperplasia with eosinophilia; angioma
sclerosing; angiomatosis; glomangioma; hemangioendothelioma;
hemangioma; hemangiopericytoma; hemangiosarcoma; lymphangioma;
lymphangiomyoma; lymphangiosarcoma; pinealoma; carcinosarcoma;
chondrosarcoma; cystosarcoma phyllodes; fibrosarcoma;
hemangiosarcoma; leiomyosarcoma; leukosarcoma; liposarcoma;
lymphangiosarcoma; myosarcoma; myxosarcoma; ovarian carcinoma;
rhabdomyosarcoma; sarcoma; neoplasms; nerofibromatosis; and
cervical dysplasia.
[0170] In a particular embodiment the present disclosure provides
methods for treating colon cancer, colorectal cancer, gastric
cancer, thyroid cancer, lung cancer, leukemia, pancreatic cancer,
melanoma, multiple melanoma, brain cancer, primary and secondary
CNS cancer, including malignant glioma and glioblastoma, renal
cancer, prostate cancer, including castration-resistant prostate
cancer, ovarian cancer, or breast cancer, including triple
negative, HER2 positive, and hormone receptor positive breast
cancers. According to such a method, a therapeutically effective
amount of at least one compound according to Formula I or a
stereoisomer, tautomer or pharmaceutically acceptable salt thereof
can be administered to a subject who has been diagnosed with a cell
proliferative disease, such as a cancer. Alternatively, a
pharmaceutical composition comprising at least one compound
according to Formula I or a stereoisomer, tautomer or
pharmaceutically acceptable salt thereof can be administered to a
subject who has been diagnosed with cancer.
[0171] In certain embodiments the compounds in accordance with the
invention are administered to a subject with cancer in conjunction
with other conventional cancer therapies such as radiation
treatment or surgery. Radiation therapy is well-known in the art
and includes X-ray therapies, such as gamma-irradiation, and
radiopharmaceutical therapies.
[0172] In certain embodiments the inventive Mnk inhibitor compounds
are used with at least one anti-cancer agent. Anti-cancer agents
include chemotherapeutic drugs. A chemotherapeutic agent includes,
but is not limited to, an inhibitor of chromatin function, a
topoisomerase inhibitor, a microtubule inhibiting drug, a DNA
damaging agent, an antimetabolite (such as folate antagonists,
pyrimidine analogs, purine analogs, and sugar-modified analogs), a
DNA synthesis inhibitor, a DNA interactive agent (such as an
intercalating agent), and a DNA repair inhibitor.
[0173] Illustrative chemotherapeutic agents include, without
limitation, the following groups: anti-metabolites/anti-cancer
agents, such as pyrimidine analogs (5-fluorouracil, floxuridine,
capecitabine, gemcitabine and cytarabine) and purine analogs,
folate antagonists and related inhibitors (mercaptopurine,
thioguanine, pentostatin and 2-chlorodeoxyadenosine (cladribine));
antiproliferative/antimitotic agents including natural products
such as vinca alkaloids (vinblastine, vincristine, and
vinorelbine), microtubule disruptors such as taxane (paclitaxel,
docetaxel), vincristin, vinblastin, nocodazole, epothilones and
navelbine, epidipodophyllotoxins (etoposide, teniposide), DNA
damaging agents (actinomycin, amsacrine, anthracyclines, bleomycin,
busulfan, camptothecin, carboplatin, chlorambucil, cisplatin,
cyclophosphamide, Cytoxan, dactinomycin, daunorubicin, doxorubicin,
epirubicin, hexamethylmelamineoxaliplatin, iphosphamide, melphalan,
merchlorehtamine, mitomycin, mitoxantrone, nitrosourea, plicamycin,
procarbazine, taxol, taxotere, temozolamide, teniposide,
triethylenethiophosphoramide and etoposide (VP 16)); antibiotics
such as dactinomycin (actinomycin D), daunorubicin, doxorubicin
(adriamycin), idarubicin, anthracyclines, mitoxantrone, bleomycins,
plicamycin (mithramycin) and mitomycin; enzymes (L-asparaginase
which systemically metabolizes L-asparagine and deprives cells
which do not have the capacity to synthesize their own asparagine);
antiplatelet agents; antiproliferative/antimitotic alkylating
agents such as nitrogen mustards (mechlorethamine, cyclophosphamide
and analogs, melphalan, chlorambucil), ethylenimines and
methylmelamines (hexamethylmelamine and thiotepa), alkyl
sulfonates-busulfan, nitrosoureas (carmustine (BCNU) and analogs,
streptozocin), trazenes--dacarbazinine (DTIC);
antiproliferative/antimitotic antimetabolites such as folic acid
analogs (methotrexate); platinum coordination complexes (cisplatin,
carboplatin), procarbazine, hydroxyurea, mitotane,
aminoglutethimide; hormones, hormone analogs (estrogen, tamoxifen,
goserelin, bicalutamide, nilutamide) and aromatase inhibitors
(letrozole, anastrozole); anticoagulants (heparin, synthetic
heparin salts and other inhibitors of thrombin); fibrinolytic
agents (such as tissue plasminogen activator, streptokinase and
urokinase), aspirin, dipyridamole, ticlopidine, clopidogrel,
abciximab; antimigratory agents; antisecretory agents (breveldin);
immunosuppressives (cyclosporine, tacrolimus (FK-506), sirolimus
(rapamycin), azathioprine, mycophenolate mofetil); anti-angiogenic
compounds (TNP470, genistein) and growth factor inhibitors
(vascular endothelial growth factor (VEGF) inhibitors, fibroblast
growth factor (FGF) inhibitors); angiotensin receptor blocker;
nitric oxide donors; anti-sense oligonucleotides; antibodies
(trastuzumab, rituximab); chimeric antigen receptors; cell cycle
inhibitors and differentiation inducers (tretinoin); mTOR
inhibitors, topoisomerase inhibitors (doxorubicin (adriamycin),
amsacrine, camptothecin, daunorubicin, dactinomycin, eniposide,
epirubicin, etoposide, idarubicin, irinotecan (CPT-11) and
mitoxantrone, topotecan, irinotecan), corticosteroids (cortisone,
dexamethasone, hydrocortisone, methylpednisolone, prednisone, and
prenisolone); growth factor signal transduction kinase inhibitors;
mitochondrial dysfunction inducers, toxins such as Cholera toxin,
ricin, Pseudomonas exotoxin, Bordetella pertussis adenylate cyclase
toxin, or diphtheria toxin, and caspase activators; and chromatin
disruptors.
[0174] In certain embodiments an Mnk inhibitor in accordance with
the present invention is used simultaneously, in the same
formulation or in separate formulations, or sequentially with an
additional agent(s) as part of a combination therapy regimen.
[0175] Mnk inhibitors according to Formula I including their
corresponding salts and pharmaceutically acceptable compositions of
Formula I compounds are also effective as therapeutic agents for
treating or preventing cytokine mediated disorders, such as
inflammation in a patient, preferably in a human. In one
embodiment, a compound or composition in accordance with the
invention is particularly useful for treating or preventing a
disease selected from chronic or acute inflammation, chronic
inflammatory arthritis, rheumatoid arthritis, psoriasis, COPD,
inflammatory bowel disease, septic shock, Crohn's disease,
ulcerative colitis, multiple sclerosis and asthma.
[0176] The inventive compounds their corresponding salts and
pharmaceutically acceptable compositions are candidate therapeutics
for treating brain related disorders which include without
limitation autism, Fragile X-syndrome, Parkinson's disease and
Alzheimer's disease. Treatment is effected by administering to a
subject in need of treatment a Formula I compound, its
pharmaceutically acceptable salt form, or a pharmaceutically
acceptable composition of a Formula I compound or its salt.
[0177] The invention also supports the use of the inventive
compounds or a pharmaceutically acceptable formulation of the
inventive compound as an inhibitor of Mnk activity. Such inhibition
is achieved by contacting a cell expressing Mnk with a compound or
a pharmaceutically acceptable formulation, to lower or inhibit Mnk
activity, to provide therapeutic efficacy for a Mnk dependent
condition in a mammal in need thereof.
[0178] Therapeutically effective dosages of a compound according to
Formula I or a composition of a Formula I compound will generally
range from about 1 to 2000 mg/day, from about 10 to about 1000
mg/day, from about 10 to about 500 mg/day, from about 10 to about
250 mg/day, from about 10 to about 100 mg/day, or from about 10 to
about 50 mg/day. The therapeutically effective dosages may be
administered in one or multiple doses. It will be appreciated,
however, that specific doses of the compounds of the invention for
any particular patient will depend on a variety of factors such as
age, sex, body weight, general health condition, diet, individual
response of the patient to be treated, time of administration,
severity of the disease to be treated, the activity of particular
compound applied, dosage form, mode of application and concomitant
medication. The therapeutically effective amount for a given
situation will readily be determined by routine experimentation and
is within the skills and judgment of the ordinary clinician or
physician. In any case the compound or composition will be
administered at dosages and in a manner which allows a
therapeutically effective amount to be delivered based upon
patient's unique condition.
General Synthetic Methods
Method 1:
##STR00005##
[0180] The formation of I is accomplished by reacting compound II
(P.sup.1 is an optional protecting group) with compound III (X is a
leaving group, such as halogen, --OTf, --OTs or --OMs, and P.sup.2
is an optional protecting group) under the Buchwald-Hartwig
conditions (such as palladium catalyst, ligand, base, solvent and
heat), followed by de-protection and/or further functional group
manipulation if necessary.
[0181] Alternatively, coupling of compound II (P.sup.1 is an
optional protecting group) and compound III (X is a leaving group,
such as halogen, --OTf, --OTs or --OMs, and P.sup.2 is an optional
protecting group) is also accomplished under the copper-mediated
Ullmann type conditions (such as copper(I) iodide, base, solvent,
heat), followed by de-protection and/or further functional group
manipulation if necessary.
Method 2:
##STR00006##
[0183] The formation of I is also accomplished by reacting compound
II (P.sup.1 is an optional protecting group) with compound IV (R is
a hydrogen or alkyl group and P.sup.2 is an optional protecting
group) under the Chan-Lam conditions (such as copper(II) acetate,
oxygen, base, solvent, heat), followed by de-protection and/or
further functional group manipulation if necessary.
Method 3:
##STR00007##
[0185] Additionally, the formation of I is also accomplished by
reacting compound II (P.sup.1 is an optional protecting group) with
compound V (P.sup.2 is an optional protecting group) under the
typical nucleophilic aromatic substitution conditions (such as
solvent, heat), followed by de-protection and/or further functional
group manipulation if necessary.
Method 4:
##STR00008##
[0187] The formation of intermediate VII is accomplished by
exposing compound VI (when P is an optional protecting group) to an
alkyl halide under basic conditions (such as sodium hydride in
tetrahydrofuran), followed by de-protection and/or further
functional group manipulation if necessary.
Method 5:
##STR00009##
[0189] The formation of IX is accomplished by exposing VIII (when P
is an optional protecting group) to the Wittig olefination
conditions (such as Ph.sub.3P.dbd.CR.sup.9aR.sup.9b, solvent and
heat), followed by de-protection and/or further functional group
manipulation if necessary.
Synthesis of Formula I Compounds
[0190] The following examples are provided for purpose of
illustration and not limitation.
Example 1
Synthesis of 5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(Cpd. No. 1F)
##STR00010##
[0191] Synthesis of
2-(4-methoxybenzyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(3)
[0192] Procedure A: To a solution of
5-bromo-2-(4-methoxybenzyl)isoindolin-1-one (1, 0.5 g, 1.5 mmol) in
dioxane (10 mL) was added 7H-pyrrolo[2,3-d]pyrimidine (2, 0.27 g,
2.25 mmol) and potassium tert-butoxide (0.51 g, 4.52 mmol) followed
by the addition of XantPhos (0.087 g, 0.15 mmol). The reaction
mixture was degassed with argon for 15 min.
Tris(dibenzylideneacetone)dipalladium(0) (0.14 g, 0.15 mmol) was
then added and the reaction mixture was heated at 90.degree. C. and
maintained at that temperature for 12 h.
[0193] Following heating, the reaction mixture was cooled and
concentrated under reduced pressure. The concentrated reaction
mixture was extracted in ethyl acetate. The organic layer was
separated, dried over sodium sulphate, filtered and concentrated
under reduced pressure. The residue obtained was purified by silica
gel (100-200 mesh) column chromatography using 5% methanol in
dichloromethane as eluent so as to afford
2-(4-methoxybenzyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(3). Yield: 0.21 g, 38%; MS (ESI) m/z 371[M+1].sup.+.
Synthesis of 5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(Cpd. No. 1F)
[0194] Procedure B: A solution of
2-(4-methoxybenzyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(3, 0.21 g, 0.55 mmol) in trifluoroacetic acid (10 mL) was heated
at 95.degree. C. for 12 h. Following heating the reaction mixture
was concentrated under reduced pressure and neutralized with
saturated solution of sodium bicarbonate. The residue thus obtained
was filtered, washed with water and then with hexane and diethyl
ether. The solid was dried to afford
5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 1F).
Yield: 0.08 g, 58%; MS (ESI) m/z 251[M+1].sup.+; .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.16 (s, 1H), 8.92 (s, 1H), 8.68 (s,
1H), 8.21-8.11 (m, 2H), 8.04 (d, J=8.2 Hz, 1H), 7.85 (d, J=8.2 Hz,
1H), 6.94 (d, J=3.8 Hz, 1H), 4.49 (s, 2H).
Example 2
Synthesis of
3-methyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd.
No. 2F)
##STR00011##
[0195] Synthesis of
5-bromo-2-(4-methoxybenzyl)-3-methylisoindolin-1-one (2)
[0196] To a stirring solution of
5-bromo-2-(4-methoxybenzyl)isoindolin-1-one (1, 1 g, 3 mmol) in
tetrahydrofuran (10 mL) at 0.degree. C., was added sodium hydride
(0.14 g, 3.62 mmol) in small portions. The resultant reaction
mixture was allowed warm to room temperature and stirred at room
temperature for an additional 10 min. Iodomethane (0.64 g, 4.51
mmol) was then added and the reaction mixture was refluxed for 1 h,
then quenched with water and extracted with ethyl acetate. The
organic layer was separated, dried over sodium sulphate and
concentrated under reduced pressure. The residue thus obtained was
purified by silica gel column chromatography using 20% ethyl
acetate in hexane as eluent to afford
5-bromo-2-(4-methoxybenzyl)-3-methylisoindolin-1-one (2). Yield:
0.3 g, 29%; MS (ESI) m/z 347[M+1].sup.+.
Synthesis of
2-(4-methoxybenzyl)-3-methyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindoli-
n-1-one (4)
[0197] The synthesis of intermediate 4 was carried out according to
the general protocol described in Procedure A. Yield: 0.1 g, 45%;
MS (ESI) m/z 385[M+1].sup.+.
Synthesis of
3-methyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd.
No. 2F)
[0198] The synthesis of compound 2F was carried out according to
the general protocol described in Procedure B. Yield: 0.05 g, 74%;
MS (ESI) m/z 265[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.16 (s, 1H), 8.91 (s, 1H), 8.77 (s, 1H), 8.15 (d, J=3.8
Hz, 2H), 8.07 (dd, J=8.1, 2.0 Hz, 1H), 7.82 (d, J=8.2 Hz, 1H), 6.94
(d, J=3.8 Hz, 1H), 4.73 (q, J=6.7 Hz, 1H), 1.44 (d, J=6.7 Hz,
3H).
Example 3
Synthesis of
3-methyl-5-(5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)isoindolin-1-one
(Cpd. No. 3)
##STR00012##
[0199] Synthesis of
3-methyl-5-(5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)isoindolin-1-one
(Cpd. No. 3)
[0200] To a solution of
3-methyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (1,
0.07 g, 0.26 mmol) in methanol (5 mL), was added 10% palladium on
carbon (70 mg). The reaction mixture was allowed to stir at room
temperature under an atmosphere of hydrogen for 48 h, then filtered
through celite and washed with methanol. The filtrate was
concentrated under reduced pressure and the residue was purified by
flash chromatography to afford
3-methyl-5-(5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)isoindolin-1-one
(Cpd. No. 3). Yield: 0.017 g, 25%; MS (ESI) m/z 267[M+1].sup.+;
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.57 (s, 1H), 8.49 (s,
1H), 8.24 (d, J=1.4 Hz, 1H), 8.09-8.01 (m, 2H), 7.65 (d, J=8.2 Hz,
1H), 4.63 (q, J=6.7 Hz, 1H), 4.21-4.17 (m, 2H), 3.20 (t, J=8.7 Hz,
2H), 1.38 (d, J=6.6 Hz, 3H).
Example 4
Synthesis of 5-(1H-pyrazolo[4,3-c]pyridin-1-yl)isoindolin-1-one
(Cpd. No. 4)
##STR00013##
[0201] Synthesis of
2-(4-methoxybenzyl)-5-(1H-pyrazolo[4,3-c]pyridin-1-yl)isoindolin-1-one
(3)
[0202] To a solution of 5-bromo-2-(4-methoxybenzyl)isoindolin-1-one
(1, 0.25 g, 2.09 mmol) in dimethyl sulfoxide (5 mL), was added
1H-pyrazolo[4,3-c]pyridine (2, 0.84 g, 2.51 mmol), and copper(I)
iodide (0.08 g, 0.42 mmol) followed by the addition of cesium
carbonate (1.35 g, 4.18 mmol). The reaction mixture was heated at
120.degree. C. for 20 h then diluted with ethyl acetate and
filtered through celite. The filtrate was evaporated under reduced
pressure and the residue was purified by silica gel column
chromatography using 10% methanol in dichloromethane as eluent to
afford
2-(4-methoxybenzyl)-5-(1H-pyrazolo[4,3-c]pyridin-1-yl)isoindolin-1-one
(3). Yield: 0.5 g, 64%; MS (ESI) m/z 371[M+1].sup.+.
Synthesis of 5-(1H-pyrazolo[4,3-c]pyridin-1-yl)isoindolin-1-one
(Cpd. No. 4)
[0203] The synthesis of compound 4 was carried out according to the
general protocol described in Procedure B. Yield: 0.05 g, 15%; MS
(ESI) m/z 251[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.26 (s, 1H), 8.68 (d, J=3.6 Hz, 2H), 8.54 (d, J=6.1 Hz,
1H), 8.07-8.02 (m, 1H), 8.00-7.84 (m, 3H), 4.50 (s, 2H).
Example 5
Synthesis of 2-(1H-pyrrolo
[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one
(Cpd. No. 5)
##STR00014##
[0204] Synthesis of
6-(4-methoxybenzyl)-2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrr-
olo[3,4-b]pyridin-5-one (3)
[0205] The synthesis of intermediate 3 was carried out according to
the general protocol described in Procedure A. Yield: 0.15 g, 23%;
MS (ESI) m/z 371.05[M+1].sup.+.
Synthesis of
2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5--
one (Cpd. No. 5)
[0206] The synthesis of compound 5 was carried out according to the
general protocol described in Procedure B. Yield: 0.028 g, 27%; MS
(ESI) m/z 251[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.95 (s, 1H), 8.78 (s, 1H), 8.47-8.37 (m, 2H), 8.26 (dd,
J=6.1, 2.4 Hz, 2H), 7.93 (d, J=8.4 Hz, 1H), 6.98 (d, J=3.5 Hz, 1H),
4.55 (s, 2H).
Example 6
Synthesis of 6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoquinoline (Cpd.
No. 6)
##STR00015##
[0207] Synthesis of 6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoquinoline
(Cpd. No. 6)
[0208] The synthesis of compound 6 was carried out according to the
general protocol described in Procedure A. Yield: 0.042 g, 13%; MS
(ESI) m/z 247[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.39 (s, 1H), 9.19 (s, 1H), 8.96 (s, 1H), 8.60-8.50 (m,
2H), 8.39-8.22 (m, 3H), 7.93 (d, J=5.8 Hz, 1H), 6.99 (d, J=3.8 Hz,
1H).
Example 7
Synthesis of 5-(4-methyl-7H-pyrrolo [2, 3-d] pyrimidin-7-yl)
isoindolin-1-one (Cpd. No. 7)
##STR00016##
[0209] Synthesis of 2-(4-methoxybenzyl)-5-(4-methyl-7H-pyrrolo [2,
3-d] pyrimidin-7-yl) isoindolin-1-one (3)
[0210] Procedure C: A solution of
4-methyl-7H-pyrrolo[2,3-d]pyrimidine (1, 0.4 g, 3.0 mmol),
5-bromo-2-(4-methoxybenzyl) isoindolin-1-one (2, 1.0 g, 4.5 mmol)
and potassium phosphate (1.91 g, 9.0 mmol) in 1, 4-dioxane (15 ml)
was degassed with nitrogen for 10 min. Copper (I) iodide (0.28 g,
1.5 mmol) and trans-1, 2-diaminocyclohexane (0.17 g, 1.5 mmol) were
added and the reaction was refluxed at 90.degree. C. for 16 h.
Progress of the reaction was monitored by TLC. After completion,
solvent was removed under reduced pressure. The reaction mixture
was diluted with water and extracted twice with ethyl acetate. The
organic layer was separated, dried over sodium sulphate, filtered
and concentrated under reduced pressure to afford
2-(4-methoxybenzyl)-5-(4-methyl-7H-pyrrolo [2, 3-d] pyrimidin-7-yl)
isoindolin-1-one (3) as a yellow solid. Yield: 0.55 g, 47%; MS
(ESI) m/z 385.12[M+1].sup.+.
Synthesis of 5-(4-methyl-7H-pyrrolo [2, 3-d] pyrimidin-7-yl)
isoindolin-1-one (Cpd. No. 7)
[0211] The synthesis of compound 7 was carried out according to the
general protocol described in Procedure B. White solid; Yield:
0.058 g, 21%; MS (ESI) m/z 265.07[M+1].sup.+; .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.76 (s, 1H), 8.66 (s, 1H), 8.17 (s, 1H),
8.07 (d, J=3.6 Hz, 1H), 8.03 (d, J=8.4 Hz, 1H), 7.84 (d, J=8.0 Hz,
1H), 6.99 (d, J=4.0 Hz, 1H), 4.48 (s, 2H), 2.73 (s, 3H).
Example 8
Synthesis of
5-(5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindoli-
n-1-one (Cpd. No. 8)
##STR00017## ##STR00018##
[0212] Synthesis of
5-bromo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine
(2)
[0213] To a solution of 5-bromo-7H-pyrrolo[2,3-d]pyrimidine (1, 0.9
g, 4.54 mmol) in tetrahydrofuran (10 mL) at 0.degree. C., was added
sodium hydride (0.27 g, 6.81 mmol, 60% in hexane). The reaction
mixture was allowed to stir at 0.degree. C. for 20 min.
Chloromethyl 2-trimethylsilylethyl ether (0.9 g, 5.44 mmol) was
then added and the reaction mixture was stirred at 0.degree. C. for
an additional 30 min., and then quenched with water. The solvent
was removed under reduced pressure and the residue was purified by
silica gel column chromatography using 5% methanol in
dichloromethane to afford
5-bromo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine
(2). Yield: 1.3 g, 86%; MS (ESI) m/z 328[M+1].sup.+.
Synthesis of
5-(1-methyl-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrr-
olo[2,3-d]pyrimidine (4)
[0214] Procedure D: To a solution of
5-bromo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine
(2, 0.5 g, 1.51 mmol) in 1,4-dioxane and water (15 mL, 4:1), was
added
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(3, 0.41 g, 1.97 mmol) and sodium carbonate (0.48 g, 4.53 mmol).
The reaction mixture was degassed with argon for 15 min., and then
[1,1-bis(diphenylphosphino)ferrocene] dichloropalladium (0.18 g,
0.22 mmol) was added and the reaction mixture heated at 90.degree.
C. for 16 h. After heating, the reaction mixture was diluted with
ethyl acetate and filtered through celite. The filtrate was
evaporated under reduced pressure and the residue was purified by
silica gel column chromatography using 5% methanol in
dichloromethane as eluent to afford
5-(1-methyl-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrr-
olo[2,3-d]pyrimidine (4). Yield: 0.7 g, 70%; MS (ESI) m/z
330[M+1].sup.+.
Synthesis of
5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine (5)
[0215] To a solution of
5-(1-methyl-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)
methyl)-7H-pyrrolo[2,3-d]pyrimidine (4, 0.7 g, 2.12 mmol) in
dichloromethane (10 mL), was added cold trifluoroacetic acid (10
mL) at 0.degree. C. The reaction mixture was warmed and allowed to
stir at room temperature for 16 h, following which it was
concentrated under reduced pressure. The residue thus obtained was
diluted with acetonitrile and aqueous ammonia and stirred at room
temperature for 1 h. After stirring the reaction mixture was
concentrated under reduced pressure and the residue purified by
neutral silica gel (100-200 mesh) column chromatography using 5%
methanol in dichloromethane as eluent to afford
5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine (5).
Yield: 0.42 g, 98%.
Synthesis of
2-(4-methoxybenzyl)-5-(5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyri-
midin-7-yl)isoindolin-1-one (7)
[0216] The synthesis of intermediate 7 was carried out according to
the general protocol described in Procedure A. Yield: 0.3 g, 33%;
MS (ESI) m/z 451[M+1].sup.+.
Synthesis of
5-(5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindoli-
n-1-one (Cpd. No. 8)
[0217] Procedure E: To a solution of
2-(4-methoxybenzyl)-5-(5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyri-
midin-7-yl)isoindolin-1-one (7, 0.05 g, 0.11 mmol) in toluene (2
mL), was added triflic acid (0.2 mL, 0.44 mmol) at room temperature
and the reaction mixture was heated in a microwave at 140.degree.
C. for 15 min. Following heating the reaction mixture was
concentrated under reduced pressure and neutralized using a
saturated solution of sodium bicarbonate. The precipitated solid
was filtered, washed with dichloromethane, followed by pentane,
methanol and diethyl ether. The washed residue was dried to afford
5-(5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindoli-
n-1-one (Cpd. No. 8). Yield: 0.03 g, 14%; MS (ESI) m/z
331[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.51
(s, 1H), 9.03 (s, 1H), 8.70 (s, 1H), 8.49 (s, 1H), 8.39 (s, 1H),
8.22 (s, 1H), 8.12-8.02 (m, 2H), 7.89 (d, J=8.2 Hz, 1H), 4.50 (s,
2H), 3.93 (s, 3H).
Example 9
Synthesis of
5-(6,7-dihydropyrido[2,3-d]pyrimidin-8(5H)-yl)-3-methylisoindolin-1-one
(Cpd. No. 9)
##STR00019##
[0218] Synthesis of
5-(6,7-dihydropyrido[2,3-d]pyrimidin-8(5H)-yl)-2-(4-methoxybenzyl)-3-meth-
ylisoindolin-1-one (3)
[0219] The synthesis of intermediate 3 was carried out according to
the general protocol described in Procedure A. Yield: 0.18 g, 51%;
MS (ESI) m/z 401[M+1].sup.+.
Synthesis of
5-(6,7-dihydropyrido[2,3-d]pyrimidin-8(5H)-yl)-3-methylisoindolin-1-one
(Cpd. No. 9)
[0220] The synthesis of compound 9 was carried out according to the
general protocol described in Procedure E. Yield: 0.03 g, 43%; MS
(ESI) m/z 281[M+1].sup.+; .sup.1H NMR (400 MHz, Methanol-d.sub.4)
.delta. 8.27 (s, 1H), 8.06 (s, 1H), 7.83 (d, J=8.1 Hz, 1H), 7.59
(d, J=1.8 Hz, 1H), 7.49 (dd, J=8.1, 1.8 Hz, 1H), 4.88 (t, J=2.5 Hz,
1H), 3.95-3.91 (m, 2H), 2.91 (t, J=6.0 Hz, 2H), 2.16 (m, 2H), 1.49
(d, J=6.7 Hz, 3H), 1.31 (d, J=14.1 Hz, 1H).
Example 10
Synthesis of
4-(1-oxoisoindolin-5-yl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one
(Cpd. No. 10)
##STR00020##
[0221] Synthesis of tert-butyl
4-chloro-1-oxo-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate
(2)
[0222] To a solution of
4-chloro-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (1, 150 mg,
0.89 mmol) in tetrahydrofuran (1 mL) was added
4-(dimethylamino)pyridine (11 mg, 0.09 mmol) and di-tert-butyl
dicarbonate (194 mg, 0.89 mmol). The reaction was stirred at room
temperature for 30 min. The resulting mixture was concentrated and
purified via column chromatography (silica, ethyl
acetate/hexanes=0-20%) to afford tert-butyl
4-chloro-1-oxo-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate
(2) as an off-white solid. Yield: 186 mg, 78%.
Synthesis of tert-butyl
4-(2-(tert-butoxycarbonyl)-1-oxoisoindolin-5-yl)-1-oxo-1,3-dihydro-2H-pyr-
rolo[3,4-c]pyridine-2-carboxylate (4)
[0223] The synthesis of intermediate 4 was carried out according to
the general protocol described in Procedure D. Yield: 58 mg,
26%.
Synthesis of
4-(1-oxoisoindolin-5-yl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one
(Cpd. No. 10)
[0224] To a suspension of tert-butyl
4-(2-(tert-butoxycarbonyl)-1-oxoisoindolin-5-yl)-1-oxo-1,3-dihydro-2H-pyr-
rolo[3,4-c]pyridine-2-carboxylate (4, 39 mg, 0.08 mmol) in methanol
(4 mL) was added a 4 M solution of hydrogen chloride in dioxane (4
mL, 16 mmol). The reaction was stirred at room temperature for 90
min, and then at 50.degree. C. for 30 min. Upon cooling, the
mixture was concentrated and triturated with ethyl acetate to
afford
4-(1-oxoisoindolin-5-yl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one
(Cpd. No. 10). Yield: 15 mg, 59%. .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 9.20 (s, 1H), 8.88 (d, J=4.8 Hz, 1H), 8.70
(s, 1H), 8.14 (d, J=0.6 Hz, 1H), 8.06 (dd, J=1.5, 7.5 Hz, 1H), 7.81
(d, J=8.1 Hz, 1H), 7.73 (d, J=4.5 Hz, 1H), 4.79 (s, 2H), 4.47 (s,
2H).
Example 11
Synthesis of
6-(1H-pyrrolo[3,2-c]pyridin-1-yl)-1H-pyrrolo[3,4-c]pyridin-3(2H)-one
(Cpd. No. 11)
##STR00021##
[0225] Synthesis of 6-chloro-2-(4-methoxybenzyl)-1H-pyrrolo
[3,4-c]pyridin-3(2H)-one (3)
[0226] To a stirring solution of 2,2,6,6-tetramethylpiperidine (10
g, 71.2 mmol) in tetrahydrofuran (10 mL) was added n-butyl lithium
(44 mL, 71.2 mmol) at -78.degree. C. To this solution were added
2-((4-methoxybenzyl)amino)acetonitrile (1, 4 g, 17.8 mmol) in
tetrahydrofuran (15 mL) followed by the addition of
(6-chloropyridin-3-yl)(piperidin-1-yl)methanone (2, 3.76 g, 21.38
mmol) in tetrahydrofuran (15 mL) at -78.degree. C. Stirring of the
reaction mixture was continued at -78.degree. C. for 7 h. The
progress of reaction was monitored by TLC. After complete
consumption of starting material, reaction was quenched with
ammonium chloride solution and extracted with ethyl acetate. The
organic layer was separated, washed with brine, dried over sodium
sulfate and concentrated under reduced pressure to a crude residue,
which was purified by column chromatography to afford
6-chloro-2-(4-methoxybenzyl)-1H-pyrrolo [3,4-c]pyridin-3(2H)-one
(3). Yield: 1.2 g, 23%; MS (ESI) m/z 289[M+1].sup.+.
Synthesis of
2-(4-methoxybenzyl)-6-(1H-pyrrolo[3,2-c]pyridin-1-yl)-1H-pyrrolo[3,4-c]py-
ridin-3(2H)-one (5)
[0227] The synthesis of intermediate 5 was carried according to the
general protocol described in Procedure A. Yield: 0.31 g, 34%; MS
(ESI) m/z 372 [M+1].sup.+.
Synthesis of
6-(1H-pyrrolo[3,2-c]pyridin-1-yl)-1H-pyrrolo[3,4-c]pyridin-3(2H)-one
(Cpd. No. 11)
[0228] The synthesis of compound 11 was carried out according to
the general protocol described in Procedure E. Yield: 0.045 g, 26%;
MS (ESI) m/z 251[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.96 (s, 1H), 8.89 (s, 1H), 8.78 (s, 1H), 8.40 (s, 2H),
8.23 (d, J=3.6 Hz, 1H), 8.09 (s, 1H), 6.98 (d, J=3.6 Hz, 1H), 4.56
(s, 2H).
Example 12
Synthesis of 5-(4-amino-7H-pyrrolo [2, 3-d] pyrimidin-7-yl)
isoindolin-1-one (Cpd. No. 12)
##STR00022##
[0229] Synthesis of
N-(4-methoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (2)
[0230] A mixture of 4-chloro-7H-pyrrolo [2, 3-d] pyrimidine (1, 2.0
g, 13.07 mmol), 4-methoxybenzylamine (3.6 g, 26.14 mmol) and
potassium carbonate (5.42 g, 39.21 mmol) in 1, 4-dioxane (20 ml)
was refluxed at 100.degree. C. for 16 h. Progress of the reaction
was monitored by TLC. After completion, solvent was removed under
reduced pressure, the reaction mixture was diluted with water and
extracted with ethyl acetate twice. The organic layer was again
washed with brine, separated, dried over sodium sulphate and
concentrated under reduced pressure. The residue was finally washed
with pentane to afford
N-(4-methoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (2) as an
off white solid. Yield: 0.82 g, 25%; MS (ESI) m/z
254.99[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
11.49 (s, 1H), 8.09 (s, 1H), 7.85 (t, J=6.0 Hz, 1H), 7.27 (d, J=8.8
Hz, 2H), 7.06 (s, 1H), 6.87 (d, J=8.8 Hz, 2H), 6.57 (s, 1H), 4.63
(d, J=6.0 Hz, 2H), 3.71 (s, 3H).
Synthesis of 2-(4-methoxybenzyl)-5-(4((4-methoxybenzyl)
amino)-7H-pyrrolo[2,3-d] pyrimidin-7-yl)isoindolin-1-one (4)
[0231] The synthesis of intermediate 4 was carried out according to
the general protocol described in Procedure A. Off-white solid;
Yield: 0.63 g, 40%; MS (ESI) m/z 506.35[M+1].sup.+.
Synthesis of 5-(4-amino-7H-pyrrolo [2, 3-d] pyrimidin-7-yl)
isoindolin-1-one (Cpd. No. 12)
[0232] The synthesis of compound 12 was carried out according to
the general protocol described in Procedure B. Off-white solid;
Yield: 0.07 g, 24%; MS (ESI) m/z 266.07[M+1].sup.+; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 8.61 (s, 1H), 8.15 (s, 2H), 7.99
(s, 1H), 7.79 (s, 1H), 7.66 (s, 1H), 7.21 (s, 2H), 6.82 (s, 1H),
4.45 (s, 2H).
Example 13
Synthesis of
7-(1,3-dihydrobenzo[c]thiophen-5-yl)-7H-pyrrolo[2,3-d]pyrimidine
(Cpd. No. 13)
##STR00023##
[0233] Synthesis of
7-(1,3-dihydrobenzo[c]thiophen-5-yl)-7H-pyrrolo[2,3-d]pyrimidine
(Cpd. No. 13)
[0234] To a solution of 5-bromo-1,3-dihydrobenzo[c]thiophene (1,
0.18 g, 0.81 mmol) in dimethylformamide (3.5 mL), were added
7H-pyrrolo [2,3-d]pyrimidine (2, 0.14 g, 1.22 mmol) and sodium
tert-butoxide (0.12 g, 1.2 mmol). The reaction mixture was degassed
with argon for 20 min. Trans-N,N'-dimethyl cyclohexane 1,2 diamine
(0.064 g, 0.4 mmol) and copper(I) iodide (0.030 g, 0.16 mmol) were
then added and the reaction mixture was heated at 100.degree. C.
for 16 h. After heating the reaction mixture was filtered through
celite and the filtrate was concentrated under reduced pressure.
The residue thus obtained was purified by silica gel column
chromatography using 2% methanol in dichloromethane as eluent to
afford
7-(1,3-dihydrobenzo[c]thiophen-5-yl)-7H-pyrrolo[2,3-d]pyrimidin- e
(Cpd. No. 13). Yield: 0.029 g, 12%; MS (ESI) m/z 254[M+1].sup.+;
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.13 (s, 1H), 8.87 (s,
1H), 8.03 (d, J=3.7 Hz, 1H), 7.84 (d, J=2.0 Hz, 1H), 7.74 (dd,
J=8.3, 2.1 Hz, 1H), 7.50 (d, J=8.2 Hz, 1H), 6.88 (d, J=3.7 Hz, 1H),
4.34-4.26 (m, 4H).
Example 14
Synthesis of
2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin--
5-one (Cpd. No. 14)
##STR00024##
[0235] Synthesis of ethyl
4-formyl-2-(methylthio)pyrimidine-5-carboxylate (2)
[0236] To a stirring solution of ethyl
4-methyl-2-(methylthio)pyrimidine-5-carboxylate (1, 2.6 g, 12.24
mmol) in 1,4-dioxane (65 mL), was added selenium dioxide (2.71 g,
24.49 mmol). The reaction mixture was heated at 100.degree. C. for
24 h. and progress of the reaction was monitored by TLC. After
completion of the reaction, the mixture was cooled and filtered.
The filtrate was concentrated under reduced pressure to afford
ethyl 4-formyl-2-(methylthio)pyrimidine-5-carboxylate (2). Yield:
2.5 g, 90%; MS (ESI) m/z 227[M+1].sup.+.
Synthesis of
6-(4-methoxybenzyl)-2-(methylthio)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-
-5-one (3)
[0237] To a solution of ethyl
4-formyl-2-(methylthio)pyrimidine-5-carboxylate (2, 2.5 g, 11.04
mmol) in methanol (25 mL) and dichloromethane (25 mL), was added
dropwise a solution of 4-methoxybenzylamine (1.51 g, 11.04 mmol).
The reaction mixture was stirred at room temperature for 30 min.
Sodium cyanoborohydride (1.73 g, 27.6 mmol) was then added and the
reaction mixture was stirred at room temperature for an additional
24 h. The progress of the reaction was monitored by TLC. After
complete consumption of starting material, the reaction mixture was
concentrated under reduced pressure and the residue was diluted
with water and the compound was extracted with dichloromethane. The
organic layer was washed with brine, dried over anhydrous sodium
sulfate and concentrated under reduced pressure. The residue was
purified by column chromatography to
6-(4-methoxybenzyl)-2-(methylthio)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-
-5-one (3). Yield: 1.6 g, 48%; MS (ESI) m/z 302[M+1].sup.+.
Synthesis of
6-(4-methoxybenzyl)-2-(methylsulfonyl)-6,7-dihydro-5H-pyrrolo[3,4-d]
pyrimidin-5-one (4)
[0238] To a solution of
6-(4-methoxybenzyl)-2-(methylthio)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-
-5-one (3, 1.6 g, 5.30 mmol) in dichloromethane (160 mL) at
0.degree. C., was added in small portions m-chloroperbenzoic acid
(2.74 g, 15.92 mmol) over a period of 30 min. The reaction mixture
was stirred at room temperature for 1 h. The progress of the
reaction was monitored by TLC. After complete consumption of
starting material the reaction mixture was quenched with saturated
solution of sodium bicarbonate and extracted with dichloromethane.
The organic layer was washed with brine, dried over anhydrous
sodium sulfate and concentrated under reduced pressure. The
resultant residue was purified by repeated washing with ether and
pentane to give
6-(4-methoxybenzyl)-2-(methylsulfonyl)-6,7-dihydro-5H-pyrrolo[3,4-
-d]pyrimidin-5-one (4). Yield: 0.45 g, 26%; MS (ESI) m/z
334[M+1].sup.+.
Synthesis of
6-(4-methoxybenzyl)-2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrr-
olo[3,4-d]pyrimidin-5-one (6)
[0239] To a solution of
6-(4-methoxybenzyl)-2-(methylsulfonyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrim-
idin-5-one (4, 0.2 g, 0.60 mmol) in acetonitrile (5 mL), was added
at 0.degree. C. 1H-pyrrolo[3,2-c]pyridine (5, 0.049 g, 0.42 mmol)
and the reaction mixture was stirred at room temperature for 1 h.
The progress of the reaction was monitored by TLC. After complete
consumption of starting material, the reaction mixture was
concentrated under reduced pressure and the residue was purified by
column chromatography to afford
6-(4-methoxybenzyl)-2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrr-
olo[3,4-d]pyrimidin-5-one (6). Yield: 0.1 g, 22%; MS (ESI) m/z
372[M+1].sup.+.
Synthesis of
2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin--
5-one, formic acid salt (Cpd. No. 14)
[0240] The synthesis of compound 14 was carried out according to
the general protocol described in Procedure B. Yield: 0.009 g, 13%;
MS (ESI) m/z 252.10[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.18 (s, 1H), 8.97 (s, 1H), 8.90 (s, 1H), 8.65 (d, J=5.8
Hz, 1H), 8.51-8.36 (m, 1H), 7.00 (d, J=3.7 Hz, 1H), 4.61 (s, 2H),
2.67-2.51 (m, 1H).
Example 15
Synthesis of 5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)indolin-2-one (Cpd.
No. 15)
##STR00025##
[0241] Synthesis of ethyl
2-(2-nitro-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)acetate
(3)
[0242] The synthesis of intermediate 3 was carried out according to
the general protocol described in Procedure A. Yield: 0.4 g,
35%.
Synthesis of ethyl
2-(2-amino-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)acetate
(4)
[0243] To a solution of ethyl
2-(2-nitro-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)acetate (3,
0.35 g, 1.07 mmol) in ethanol (25 mL), was added 10% palladium on
carbon (150 mg) and the reaction mixture was allowed to stir at
room temperature under an atmosphere of hydrogen for 4 h. The
reaction mixture was filtered through a celite bed and the filtrate
was concentrated under reduced pressure to afford ethyl
2-(2-amino-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)acetate (4).
Yield: 0.38 g, crude.
Synthesis of 5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)indolin-2-one (Cpd.
No. 15)
[0244] A solution of ethyl
2-(2-amino-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)acetate (4,
(0.35 g, 1.18 mmol) in ethanol (20 mL) was allowed to reflux for 16
h. The reaction mixture was concentrated under reduced pressure and
the residue was purified by neutral silica gel column
chromatography using 5% methanol in dichloromethane as eluent to
afford compound 5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)indolin-2-one
(Cpd. No. 15). The compound was lyophilised to remove the trapped
methanol and triturated with diethyl ether to remove the non polar
impurity. Yield: 0.058 g, 17%. MS (ESI) m/z 251[M+1].sup.+; .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 10.56 (s, 1H), 9.11 (s, 1H),
8.83 (s, 1H), 7.92 (d, J=3.9 Hz, 1H), 7.68 (s, 1H), 7.58 (d, J=8.3
Hz, 1H), 6.97 (d, J=8.3 Hz, 1H), 6.89-6.80 (m, 1H), 3.60 (s,
2H).
Example 16
Synthesis of 5-(9H-purin-9-yl) isoindolin-1-one (Cpd. No. 16)
##STR00026## ##STR00027##
[0245] Synthesis of
2-(4-methoxybenzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoind-
olin-1-one (2)
[0246] A mixture of 5-bromo-2-(4-methoxybenzyl) isoindolin-1-one
(1, 1.0 g, 3.02 mmol), bis(pinacolato) diboron (0.84 g, 3.32 mmol)
and potassium acetate (0.74 g, 7.55 mmol) in 1,4-dioxane (10 ml)
was degassed with argon at room temperature for 15 min. Then
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.22
g, 0.30 mmol) was added under nitrogen atmosphere and the reaction
was purged for another 10 min. The reaction was allowed to reflux
at 100.degree. C. for 18 h. Progress of the reaction was monitored
by TLC. After completion, the reaction mass was filtered through
celite and the celite bed was washed with ethyl acetate. The
combined organic layer was dried over sodium sulphate and
concentrated under reduced pressure to afford
2-(4-methoxybenzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoind-
olin-1-one (2) as a black solid. Yield: 1.2 g, crude; MS (ESI) m/z
380.27[M+1].sup.+.
Synthesis of mixture of 2-(4-methoxybenzyl)-5-(9H-purin-9-yl)
isoindolin-1-one (4) and 2-(4-methoxybenzyl)-5-(7H-purin-7-yl)
isoindolin-1-one (4a)
[0247] A stirred solution of
2-(4-methoxybenzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoind-
olin-1-one (2, 1.26 g, 3.34 mmol), 9H-purine (3, 0.2 g, 1.67 mmol)
and tetramethylethylenediamine (0.39 g, 3.34 mmol) in methanol (30
ml) and water (5 ml) was degassed with oxygen for 10 min. Copper
(II) acetate (0.31 g, 1.67 mmol) was then added and the reaction
was stirred at room temperature for 16 h under an atmosphere of
oxygen. Progress of the reaction was monitored by TLC. After
completion, solvent was removed under reduced pressure. The
reaction mixture was diluted with water and extracted twice with
ethyl acetate. The organic layer was separated, dried over sodium
sulphate and concentrated under reduced pressure. The residue was
purified via column chromatography using 5% methanol in
dichloromethane to afford a mixture of
2-(4-methoxybenzyl)-5-(9H-purin-9-yl) isoindolin-1-one (4) and
2-(4-methoxybenzyl)-5-(7H-purin-7-yl) isoindolin-1-one (4a) as a
brown solid. Yield: 0.46 gm, 36%; MS (ESI) m/z 372.05 and
372.09[M+1].sup.+.
Synthesis of 5-(9H-purin-9-yl) isoindolin-1-one (Cpd. No. 16)
[0248] The synthesis of compound 16 was carried out according to
the general protocol described in Procedure B. White solid; Yield:
0.075 g, 52%; MS (ESI) m/z 252.03[M+1].sup.+; .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.33 (s, 1H), 9.14 (s, 1H), 9.06 (s, 1H),
8.77 (s, 1H), 8.23 (s, 1H), 8.08 (d, J=8.4 Hz, 1H), 7.91 (d, J=8.0
Hz, 1H), 4.51 (s, 2H).
Example 17
Synthesis of 7-chloro-3-methyl-5-(7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 17)
##STR00028## ##STR00029##
[0249] Synthesis of methyl 4-bromo-2-(bromomethyl)-6-chlorobenzoate
(2)
[0250] To a solution of methyl 4-bromo-2-chloro-6-methylbenzoate
(1, 2.0 g, 7.6 mmol) in carbon tetrachloride (20 ml) were added
N-bromosuccinimide (1.6 g, 9.1 mmol) and
2,2'-azobis(2-methylpropionitrile) (0.25 g, 1.52 mmol). The
reaction mixture was stirred at 80.degree. C. for 12 h and progress
of the reaction was monitored by TLC. After completion, solvent was
diluted with dichloromethane and the organic layer was washed with
water and brine. Following separation, the organic layer was dried
using sodium sulphate and concentrated under reduced pressure to
afford methyl 4-bromo-2-(bromomethyl)-6-chlorobenzoate (2) as a
yellow sticky liquid. Yield: 3.9 g, crude.
Synthesis of 5-bromo-7-chloro-2-(4-methoxybenzyl) isoindolin-1-one
(3)
[0251] To a solution of methyl
4-bromo-2-(bromomethyl)-6-chlorobenzoate (2, 3.0 g, 8.8 mmol) in
N,N-dimethylformamide (25 ml) were added 4-methoxybenzylamine (1.8
g, 13.0 mmol) and triethylamine (2.67 g, 26.4 mmol). The reaction
was stirred at room temperature for 12 h and progress of reaction
was monitored by TLC. After completion, the reaction mass was
quenched with water and the desired compound was extracted from the
crude reaction mass with ethyl acetate twice. The combined organic
layers were washed with cold water and cold brine. Following
separation, the organic layer was dried using sodium sulphate and
concentrated under reduced pressure. The residue was purified via
column chromatography using 10% ethyl acetate in hexane to get
5-bromo-7-chloro-2-(4-methoxybenzyl) isoindolin-1-one (3) as a
yellow sticky liquid. Yield: 1.6 g, 50%; MS (ESI) m/z
365.97[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.77
(d, J=4.4 Hz, 2H), 7.22 (d, J=8.8 Hz, 2H), 6.91 (d, J=8.4 Hz, 2H),
4.61 (s, 2H), 4.29 (s, 2H), 3.73 (s, 3H).
Synthesis of mixture of
5-bromo-7-chloro-2-(4-methoxybenzyl)-3-methylisoindolin-1-one (4)
and
5-bromo-7-chloro-2-(4-methoxybenzyl)-3,3-dimethylisoindolin-1-one
(4')
[0252] To a solution of 5-bromo-7-chloro-2-(4-methoxybenzyl)
isoindolin-1-one (3, 1.5 g, 4.1 mmol) in dry tetrahydrofuran (20
mL) at 0.degree. C. was added solid sodium bis(trimethylsilyl)amide
(0.89 g, 4.92 mmol). The reaction mass was stirred at 0.degree. C.
for 15 min. Iodomethane (5.8 g, 41.0 mmol) was then added and the
reaction mixture was stirred at 0.degree. C. for an additional 3 h.
Progress of the reaction was monitored by TLC. After consumption of
starting material, the reaction mixture was quenched with water and
extracted twice with ethyl acetate. The organic layer was washed
with brine solution, dried over anhydrous sodium acetate, filtered
and concentrated under reduced pressure to get a mixture of
5-bromo-7-chloro-2-(4-methoxybenzyl)-3-methylisoindolin-1-one (4)
and
5-bromo-7-chloro-2-(4-methoxybenzyl)-3,3-dimethylisoindolin-1-one
(4') as a yellow sticky liquid. Yield: 0.85 g, crude; MS (ESI) m/z
380.02[M+1].sup.+ and 394.04[M+1].sup.+.
Synthesis of mixture of
7-chloro-2-(4-methoxybenzyl)-3-methyl-5-(7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (6) and
7-chloro-2-(4-methoxybenzyl)-3, 3-dimethyl-5-(7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (6')
[0253] The synthesis of a mixture of 6 and 6' was carried out
according to the general protocol described in Procedure C. Brown
solid; Yield: 0.45 g, crude; MS (ESI) m/z 419.27[M+1].sup.+ and
433.28[M+1].sup.+.
Synthesis of 7-chloro-3-methyl-5-(7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 17)
[0254] The synthesis of compound 17 was carried out according to
the general protocol described in Procedure B. Yield: 0.057 g, 40%;
MS (ESI) m/z 299.02[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.17 (s, 1H), 8.96 (s, 1H), 8.87 (s, 1H), 8.22 (d, J=4.0
Hz, 2H), 8.18 (s, 1H), 6.96 (d, J=3.6 Hz, 1H), 4.70 (m, 1H), 1.43
(d, J=6.4 Hz, 3H).
Example 18
Synthesis of 7-chloro-3, 3-dimethyl-5-(7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 18)
##STR00030## ##STR00031##
[0255] Synthesis of 7-chloro-3, 3-dimethyl-5-(7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 18)
[0256] The synthesis of compound 18 was carried out as described
above in Example 17. Yield: 0.025 g, 17%; MS (ESI) m/z
313.02[M+1].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.17
(s, 1H), 8.96 (s, 1H), 8.87 (s, 1H), 8.26 (s, 1H), 8.23 (d, J=3.6
Hz, 1H), 8.17 (s, 1H), 6.96 (d, J=4.0 Hz, 1H), 1.51 (s, 6H).
Example 19
Synthesis of
5-(5-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(Cpd. No. 19)
##STR00032##
[0257] Synthesis of
5-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (2)
[0258] To a solution of 5-(7H-pyrrolo [2, 3-d] pyrimidin-7-yl)
isoindolin-1-one (1, 1.3 g, 5.17 mmol) in N,N-dimethylformamide (15
ml) was added N-bromosuccinimide (1.04 g, 5.69 mmol) and the
reaction mixture was stirred at room temperature for 1 h. Progress
of the reaction was monitored by TLC. After completion, the
reaction mass was quenched with water and extracted twice with
ethyl acetate. The organic layer was then separated, dried over
sodium sulphate and concentrated under reduced pressure to afford
5-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (2) as
an off white solid. Yield: 0.85 g, 50%; MS (ESI) m/z
329.01[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.09
(s, 1H), 9.00 (s, 1H), 8.70 (s, 1H), 8.45 (s, 1H), 8.15 (s, 1H),
8.00 (d, J=8.0 Hz, 1H), 7.85 (d, J=8.0 Hz, 1H), 4.48 (s, 2H).
Synthesis of 5-(5-(pyridin-4-yl)-7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 19)
[0259] The synthesis of compound 19 was carried out according to
the general protocol described in Procedure D. Off white solid;
Yield: 0.018 g, 12%; MS (ESI) m/z 328.06[M+1].sup.+; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.65 (s, 1H), 9.02 (s, 1H), 8.88
(s, 1H), 8.72 (s, 1H), 8.66 (d, J=5.6 Hz, 2H), 8.25 (s, 1H), 8.11
(d, J=8.4 Hz, 1H), 7.97 (d, J=5.6 Hz, 2H), 7.90 (d, J=8.4 Hz, 1H),
4.52 (s, 2H).
Example 20
Synthesis of methyl
((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbama-
te (Cpd. No. 20)
##STR00033## ##STR00034##
[0260] Synthesis of ethyl
2-(6-bromo-2-(4-methoxybenzyl)-3-oxoisoindolin-1-yl)acetate (3)
[0261] To a solution of 5-bromo-2-(4-methoxybenzyl)isoindolin-1-one
(1, 1 g, 3.01 mmol) in tetrahydrofuran (25 mL) at -78.degree. C.
was added dropwise a solution of sodium bis(trimethylsilyl)amide
(552 mg, 3.01 mmol) in tetrahydrofuran (15 mL). The reaction was
stirred at -78.degree. C. for 15 min, followed by the dropwise
addition of ethyl 2-bromoacetate (2, 503 mg, 3.01 mmol) in
tetrahydrofuran (10 mL). The reaction was stirred at -78.degree. C.
for an additional 20 min before it was warmed to room temperature
gradually. The warmed reaction mixture is then poured into a half
saturated ammonium chloride solution and the aqueous solution is
extracted with ethyl acetate. The ethyl acetate layers were
combined, washed with brine, dried over magnesium sulfate, filtered
and concentrated under reduced pressure. The crude product was
purified via column chromatography (silica, ethyl
acetate/hexanes=0-10%) to afford ethyl
2-(6-bromo-2-(4-methoxybenzyl)-3-oxoisoindolin-1-yl)acetate (3).
Yield: 572 mg, 45%; MS (ESI) m/z 418.3[M-1].sup.+.
Synthesis of ethyl
2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindoli-
n-1-yl)acetate (5)
[0262] A mixture of ethyl
2-(6-bromo-2-(4-methoxybenzyl)-3-oxoisoindolin-1-yl)acetate (3, 439
mg, 1.05 mmol), 7H-pyrrolo[2,3-d]pyrimidine (4, 125 mg, 1.05 mmol),
tris(dibenzylideneacetone)dipalladium(0) (97 mg, 0.10 mmol),
XantPhos (61 mg, 0.10 mmol), and cesium carbonate (752 mg, 2.31
mmol) in 1,4-dioxane (25 mL) was purged with argon for 5 min. The
reaction was stirred at 110.degree. C. for 16 h. Upon cooling, the
reaction mixture was diluted with ethyl acetate, washed with half
saturated aqueous sodium bicarbonate solution, and then with brine.
The organic layer was dried over magnesium sulfate, filtered and
concentrated. The crude product was purified via column
chromatography (silica, methanol/dichloromethane gradient from 0-5%
to afford ethyl
2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindoli-
n-1-yl)acetate (5). Yield: 333 mg, 70%; MS (ESI) m/z
457.4[M+1].sup.+.
Synthesis of
2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindoli-
n-1-yl)acetic acid (6)
[0263] To a solution of ethyl
2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindoli-
n-1-yl)acetate (4, 333 mg, 0.73 mmol) in methanol (10 mL) and water
(5 mL) was added lithium hydroxide (52 mg, 2.19 mmol). The reaction
was stirred at room temperature for 2 h and then acidified to pH
.about.5 with 1.25 M hydrogen chloride in ethanol. The resulting
mixture was concentrated to afford
2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)is-
oindolin-1-yl)acetic acid (6). Yield: 314 mg, 100%; MS (ESI) m/z
429.2[M+1].sup.+.
Synthesis of methyl
((2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-
-1-yl)methyl)carbamate (7)
[0264] To a suspension of
2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindoli-
n-1-yl)acetic acid (6, 312 mg, 0.73 mmol) in tetrahydrofuran (10
mL) at 0.degree. C. was added triethylamine (0.41 mL, 2.92 mmol),
and ethyl chloroformate (119 mg, 1.09 mmol). The reaction was
stirred at 0.degree. C. for 1 h. A solution of sodium azide (95 mg,
1.46 mmol) in water (2 mL) was then added to the reaction mixture.
After stirring at 0.degree. C. for 5 min the reaction mixture was
warmed to room temperature and allowed to stir for an additional 2
h. The resulting mixture was poured into water and extracted with
ethyl acetate. The separated organic layer was washed with half
saturated aqueous sodium bicarbonate solution (20 mL), and brine.
It was then dried over magnesium sulfate, filtered and
concentrated. The residue thus obtained was dissolved in
dichloromethane (15 mL) and refluxed for 30 min. The reaction was
cooled to room temperature followed by the addition of methanol (1
mL). The reaction was stirred at reflux for an additional 1 h. The
resulting mixture was concentrated and purified via column
chromatography (silica, methanol/dichloromethane=0-5%) to afford
methyl
((2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-
-1-yl)methyl)carbamate (7). Yield: 181 mg, 54%; MS (ESI) m/z
458.5[M+1].sup.+.
Synthesis of methyl
((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbama-
te (Cpd. No. 20)
[0265] The synthesis of compound 20 was carried out according to
the general protocol described in Procedure B. White solid; Yield:
34 mg, 42%; MS (ESI) m/z 338.3[M+1].sup.+; .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 9.42 (s, 1H), 9.15 (s, 1H), 8.31 (d, J=3.9 Hz,
1H), 8.13 (s, 1H), 8.07-7.98 (m, 2H), 7.25 (d, J=3.6 Hz, 1H), 4.92
(s, 1H), 3.72-3.50 (m, 5H).
Example 21
Synthesis of
3-(aminomethyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
hydrochloride (Cpd. No. 21)
##STR00035##
[0266] Synthesis of tert-butyl
((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbama-
te (2)
[0267] To a solution of methyl
((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbama-
te (1, 22 mg, 0.07 mmol) in acetonitrile (5 mL) was added
iodotrimethylsilane (0.02 mL, 0.13 mmol). The reaction was stirred
at room temperature for 2 h. The resulting mixture was concentrated
and re-dissolved in dichloromethane (5 mL), followed by the
addition of di-tert-butyl dicarbonate (29 mg, 0.13 mmol). After
stirring at room temperature for 4 h, the mixture was concentrated
and purified via column chromatography (silica,
methanol/dichloromethane=0-10%) to afford tert-butyl
((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbama-
te (2). Yield: 18 mg, 73%; MS (ESI) m/z 380.2[M+1].sup.+.
Synthesis of
3-(aminomethyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
hydrochloride (Cpd. No. 21)
[0268] To a solution of tert-butyl
((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbama-
te (2, 18 mg, 0.05 mmol) in methanol (6 mL) was added 4 M hydrogen
chloride in dioxane (0.01 mL, 0.05 mmol). The reaction was stirred
at room temperature for 2 h and the resulting mixture was
concentrated and triturated with methanol and ether to afford
3-(aminomethyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
hydrochloride (Cpd. No. 21) as a white solid. Yield: 8 mg, 50%; MS
(ESI) m/z 280.4[M+1].sup.+. .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 9.50 (s, 1H), 9.22 (s, 1H), 8.39 (d, J=3.6 Hz, 1H),
8.27-8.26 (m, 1H), 8.17 (dd, J=7.8, 1.5 Hz, 1H), 8.08 (d, J=8.4 Hz,
1H), 7.33 (d, J=3.9 Hz, 1H), 5.18 (t, J=4.2 Hz, 1H), 3.70-3.51 (m,
2H).
Example 22
Synthesis of
3,7-dimethyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(Cpd. No. 22)
##STR00036##
[0269] Synthesis of 5-bromo-2-(4-methoxybenzyl)-3,
7-dimethylisoindolin-1-one (2)
[0270] To a solution of
5-bromo-2-(4-methoxybenzyl)-7-methylisoindolin-1-one (1, 2.2 g, 6.3
mmol) in tetrahydrofuran (20 mL) at 0.degree. C. was added solid
sodium bis(trimethylsilyl)amide (1.39 g, 7.6 mmol). The reaction
was stirred at 0.degree. C. for 15 min. Iodomethane (1.17 g, 8.2
mmol) was then added and reaction mixture stirred at 0.degree. C.
for an additional 4 h. Progress of the reaction was monitored by
TLC. After consumption of starting material, the reaction mixture
was quenched with water and extracted with ethyl acetate. After
separation of the organic layer from the aqueous layer, the former
was washed with brine, dried over anhydrous sodium sulfate,
filtered and concentrated under reduced pressure to afford
5-bromo-2-(4-methoxybenzyl)-3,7-dimethylisoindolin-1-one (2) as a
white solid. Yield: 1.0 g, 44%; MS (ESI) m/z 360.16[M+1].sup.+.
Synthesis of
2-(4-methoxybenzyl)-3,7-dimethyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoin-
dolin-1-one (4)
[0271] The synthesis of intermediate 4 was carried out according to
the general protocol described in Procedure C. Off white solid;
Yield: 0.41 g, 93%. MS (ESI) m/z 399.24[M+1].sup.+.
Synthesis of
3,7-dimethyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(Cpd. No. 22)
[0272] The synthesis of compound 22 was carried out according to
the general protocol described in Procedure B. Off white solid;
Yield: 0.14 g, 57%. MS (ESI) m/z 279.09[M+1].sup.+; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.150 (s, 1H), 8.916 (s, 1H), 8.639
(s, 1H), 8.110-8.119 (d, J=3.6 Hz, 1H), 7.937 (s, 1H), 7.791 (s,
1H), 6.915-6.924 (d, J=3.8 Hz, 1H), 4.67 (m, 1H), 2.685 (s, 3H),
1.397-1.414 (d, J=6.8 Hz, 3H).
Example 23
Synthesis of 7-fluoro-5-(7H-pyrrolo [2, 3-d] pyrimidin-7-yl)
isoindolin-1-one (Cpd. No. 23)
##STR00037##
[0273] Synthesis of methyl 4-bromo-2-(bromomethyl)-6-fluorobenzoate
(2)
[0274] To a solution of methyl 4-bromo-2-fluoro-6-methylbenzoate
(1, 1.8 g, 7.31 mmol) in carbon tetrachloride (100 ml) were added
N-bromosuccinimide (1.56 g, 8.78 mmol) and
2,2'-azobis(2-methylpropionitrile) (0.24 g, 1.46 mmol). The
reaction was refluxed at 80.degree. C. for 18 h and the progress
was monitored by TLC. After completion, solvent was removed under
reduced pressure to afford methyl
4-bromo-2-(bromomethyl)-6-fluorobenzoate (2) as a brown solid.
Yield: 3.9 g, crude; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.76 (s, 1H), 7.75 (s, 1H), 4.76 (s, 2H), 3.93 (s, 3H).
Synthesis of 5-bromo-7-fluoro-2-(4-methoxybenzyl) isoindolin-1-one
(3)
[0275] A solution of methyl
4-bromo-2-(bromomethyl)-6-fluorobenzoate (2, 2.2 g, 6.79 mmol),
4-methoxybenzylamine (1.87 g, 13.58 mmol) and triethylamine (2.06
g, 20.37 mmol) in N,N-dimethylformamide (20 ml) was allowed to stir
at room temperature for 48 h. Progress of the reaction was
monitored by TLC. After completion, the reaction mass was quenched
with water and the aqueous solution was extracted twice with ethyl
acetate. The ethyl acetate layer was washed with cold water then
cold brine. The ethyl acetate layer was separated, dried over
sodium sulphate and concentrated under reduced pressure. The
residue was purified via column chromatography using 26% ethyl
acetate in hexane to get 5-bromo-7-fluoro-2-(4-methoxybenzyl)
isoindolin-1-one (3) as a yellow solid. Yield: 1.7 g, 72%; MS (ESI)
m/z 350.03[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
7.65 (s, 1H), 7.62 (s, 1H), 7.22 (d, J=8.4 Hz, 2H), 6.92 (d, J=8.4
Hz, 2H), 4.60 (s, 2H), 4.33 (s, 2H), 3.72 (s, 3H).
Synthesis of 7-fluoro-2-(4-methoxybenzyl)-5-(7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (5)
[0276] The synthesis of intermediate 5 was carried out according to
the general protocol described above in Procedure C. Off-white
solid; Yield: 0.28 g, 51%; MS (ESI) m/z 389.19[M+1].sup.+; .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 9.16 (s, 1H), 8.94 (s, 1H),
8.16 (d, J=4.0 Hz, 1H), 8.08 (s, 1H), 8.03 (d, J=10.8 Hz, 1H), 7.25
(d, J=8.0 Hz, 2H), 6.95 (t, J=4.0 Hz, 2H), 6.92 (s, 1H), 4.65 (s,
2H), 4.47 (s, 2H), 3.74 (s, 3H).
Synthesis of 7-fluoro-5-(7H-pyrrolo [2, 3-d] pyrimidin-7-yl)
isoindolin-1-one (Cpd. No. 23)
[0277] 7-fluoro-2-(4-methoxybenzyl)-5-(7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (5, 0.27 g, 0.69 mmol) was
dissolved in a mixture of trifluoroacetic acid (5 ml), triflic acid
(5 ml) and dichloromethane (5 ml). The reaction was stirred at
60.degree. C. for 3 h. Progress of the reaction was monitored by
TLC. After completion, the reaction mass was quenched with water
and washed with ethyl acetate. The resulting aqueous layer was
basified with saturated aqueous sodium bicarbonate and extracted
with ethyl acetate twice. The organic layer was again washed with
brine, separated, dried over sodium sulphate and concentrated under
reduced pressure. The residue was purified via column
chromatography using 2% methanol in dichloromethane to get
7-fluoro-5-(7H-pyrrolo [2, 3-d] pyrimidin-7-yl) isoindolin-1-one
(Cpd. No. 23) as an off-white solid. Yield: 0.06 g, 32%; MS (ESI)
m/z 269.05[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.17 (s, 1H), 8.95 (s, 1H), 8.69 (s, 1H), 8.21 (d, J=3.6 Hz, 1H),
8.14 (s, 1H), 7.99 (d, J=11.2 Hz, 1H), 6.96 (d, J=3.6 Hz, 1H), 4.50
(s, 2H).
Example 24
Synthesis of 5-(5-(2-chlorophenyl)-7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 24)
##STR00038##
[0278] Synthesis of 5-(5-(2-chlorophenyl)-7H-pyrrolo [2, 3-d]
pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 24)
[0279] The synthesis of compound 24 was carried out according to
the general protocol described above in Procedure D. White solid;
Yield: 0.04 g, 18%; MS (ESI) m/z 361.01[M+1].sup.+; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.10 (s, 1H), 8.99 (s, 1H), 8.71
(s, 1H), 8.39 (s, 1H), 8.24 (s, 1H), 8.11 (d, J=6.8 Hz, 1H), 7.88
(d, J=8.0 Hz, 1H), 7.75 (d, J=8.0 Hz, 1H), 7.68 (d, J=8.0 Hz, 1H),
7.49 (m, 2H), 4.51 (s, 2H).
Example 25
Synthesis of
5-(5-(thiazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(Cpd. No. 25)
##STR00039## ##STR00040##
[0280] Synthesis of
2-(4-methoxybenzyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(3)
[0281] The synthesis of intermediate 3 was carried out according to
the general protocol described above in Procedure C. Yellow solid.
Yield: 3.0 g, 54%. MS (ESI) m/z 371.2[M+1].sup.+. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.15 (s, 1H), 8.90 (s, 1H), 8.12-8.10
(m, 2H), 8.07 (d, J=8.4 Hz, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.24 (d,
J=8.4 Hz, 2H), 6.92 (m, 3H), 4.69 (s, 2H), 4.44 (s, 2H), 3.73 (s,
3H).
Synthesis of
5-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-(4-methoxybenzyl)isoindolin-
-1-one (4)
[0282] Small portions of N-bromosuccinimide (1.15 g, 6.47 mmol)
were added to a stirring solution of
2-(4-methoxybenzyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(3, 2 g, 5.39 mmol) in N,N-dimethylformamide (30 mL), at room
temperature. The reaction mass was stirred at room temperature for
1 h. After completion, the reaction mixture was diluted with water
(50 mL) and extracted with ethyl acetate (2.times.50 mL). The
organic layers were combined, dried using magnesium sulfate,
filtered and concentrated to dryness under vacuum. The crude was
then purified by flash column chromatography using 2.5% methanol in
dichloromethane as the eluant. The desired fractions were
concentrated to dryness under vacuum to afford
5-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-(4-methoxybenzyl)isoindolin-
-1-one (4) as a brown solid. Yield: 1.2 g, 49%. MS (ESI) m/z
451.19[M+1].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.07
(s, 1H), 8.98 (s, 1H), 8.41 (s, 1H), 8.09 (s, 1H), 8.03 (d, J=7.2
Hz, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.23 (d, J=8.4 Hz, 2H), 6.92 (d,
J=8.4 Hz, 2H), 4.69 (s, 2H), 4.43 (s, 2H), 3.73 (s, 3H).
Synthesis of
2-(4-methoxybenzyl)-5-(5-(thiazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)i-
soindolin-1-one (6)
[0283] The synthesis of intermediate 6 was carried out according to
the general protocol described above in Procedure D. Yellow solid;
Yield: 0.35 g, 50%. MS (ESI) m/z 354.22[M+1].sup.+.
Synthesis of
5-(5-(thiazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
(Cpd. No. 25)
[0284] A solution containing
2-(4-methoxybenzyl)-5-(5-(thiazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)i-
soindolin-1-one (0.35 g, 0.77 mmol) in trifluoroacetic acid (5 mL),
triflic acid (5 mL) and dichloromethane (5 mL) was heated at
60.degree. C. for 16 h. After completion, the reaction mixture was
concentrated to dryness, quenched with an aqueous solution of
sodium bicarbonate until the pH is 8.0 and extracted with 10%
methanol in dichloromethane (2.times.50 mL). The organic layers
were combined, dried using magnesium sulfate and concentrated to
dryness under vacuum. The crude was then purified by prep HPLC and
the desired fractions were concentrated to dryness under vacuum to
afford
5-(5-(thiazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one
as a yellow solid. Yield: 0.020 g, 8%. MS (ESI) m/z
334.05[M+1].sup.+; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.69
(s, 1H), 9.31 (s, 1H), 8.99 (s, 1H), 8.69 (s, 2H), 8.25 (m, 2H),
8.11 (d, J=8.4 Hz, 1H), 7.88 (d, J=8.0 Hz, 1H), 4.50 (s, 2H).
Example 26
Synthesis of
2-(7-(1-oxoisoindolin-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)benzonitrile
(Cpd. No. 26)
##STR00041##
[0285] Synthesis of 2-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)benzonitrile
(3)
[0286] The synthesis of intermediate 3 was carried out according to
the general protocol described above in Procedure D. Yellow solid.
Yield: 0.75 g, crude. MS (ESI) m/z:221 [M+1].sup.+. LCMS: 44%
Synthesis of
2-[7-(1-oxoisoindolin-5-yl)pyrrolo[2,3-d]pyrimidin-5-yl]benzonitrile
(Cpd. No. 26)
[0287] The synthesis of compound 26 was carried out according to
the general protocol described above in Procedure C. White solid;
Yield: 0.025 g, 5%. MS (ESI) m/z 352.2[M+1].sup.+; .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.25 (s, 1H), 9.02 (s, 1H), 8.73 (s,
1H), 8.57 (s, 1H), 8.27 (s, 1H), 8.07-8.04 (m, 2H), 7.96-7.85 (m,
3H), 7.65-7.61 (m, 1H), 4.52 (s, 2H).
Example 27
Synthesis of
4'-chloro-6'-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclopentane-1,1'-iso-
indolin]-3'-one (Cpd. No. 27)
##STR00042##
[0288] Synthesis of
6'-bromo-4'-chloro-2'-(4-methoxybenzyl)spiro[cyclopentane-1,1'-isoindolin-
]-3'-one (3)
[0289] To a solution of
5-bromo-7-chloro-2-[(4-methoxyphenyl)methyl]isoindolin-1-one (1,
0.4 g, 1.09 mmol) in tetrahydrofuran (25 mL) at room temperature
was added sodium hydride (131 mg, 5.45 mmol). The reaction was
stirred for 30 min and then 1,4-diiodobutane (2, 1691 mg, 5.45
mmol) was added to the reaction mixture. The reaction was stirred
at room temperature for an additional 5 h. After completion, the
reaction mass was quenched with a cold saturated solution of
ammonium chloride at 0.degree. C. The residue was dissolved in
ethyl acetate (100 mL) and the organic layer was washed with water
(2.times.20 mL) then with brine (10 mL). The organics were
separated and dried using magnesium sulfate before concentration to
dryness. The crude was then purified by flash column chromatography
using 10% ethyl acetate in hexane as the eluant. The desired
fractions were concentrated to dryness under vacuum to afford
5'-bromo-7'-chloro-2'-[(4-methoxyphenyl)methyl]spiro[cyclopentane-1,3'-is-
oindoline]-1'-one as a yellow solid. Yield: 0.21 g, 45%; MS (ESI)
m/z 422.2[M+1].sup.+; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.54 (s, 1H), 7.40 (s, 1H), 7.26 (d, J=8.10 Hz, 2H), 6.83 (d,
J=8.10 Hz, 2H), 4.64 (s, 2H), 3.95 (s, 3H), 2.17-1.72 (m, 8H).
Synthesis of
4'-chloro-2'-(4-methoxybenzyl)-6'-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[-
cyclopentane-1,1'-isoindolin]-3'-one (5)
[0290] The synthesis of intermediate 5 was carried out according to
the general protocol described above in Procedure C. Brown solid;
Yield: 0.065 g, 29%; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.16 (s, 1H), 8.95 (s, 1H), 8.30-8.27 (m, 2H), 8.11 (s, 1H), 7.28
(d, J=8.04 Hz, 2H), 6.95 (d, J=3.28 Hz, 1H), 6.90 (d, J=8.52 Hz,
2H), 4.64 (s, 2H), 3.71 (s, 3H), 2.01-1.92 (m, 8H).
Synthesis of
4'-chloro-6'-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclopentane-1,1'-iso-
indolin]-3'-one (Cpd. No. 27)
[0291] Procedure F: A solution of
7'-chloro-2'-[(4-methoxyphenyl)methyl]-5'-pyrrolo[2,3-d]pyrimidin-7-yl-sp-
iro[cyclopentane-1,3'-isoindoline]-1'-one (5, 0.06 g, 0.13 mmol) in
dichloromethane (5 mL) and trifluoroacetic acid (10 mL) was heated
at 60.degree. C. for 48 h. After completion, the reaction mixture
was cooled to room temperature and concentrated. The crude was
co-evaporated with dichloromethane and then liquid ammonia was
added to neutralize the reaction mass. The crude was then purified
by flash column chromatography using a gradient (2-10%)methanol in
dichloromethane. The desired column fractions were concentrated to
dryness under vacuum to afford
4'-chloro-6'-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclopentane-1,1'-iso-
indolin]-3'-one (Cpd. No. 27) as a brown solid. Yield: 0.025 g,
56%; MS (ESI) m/z 338.87[M+1].sup.+; .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.16 (d, J=8.0 Hz, 2H), 8.96 (s, 1H), 8.25
(d, J=6.48 Hz, 2H), 8.13 (d, J=1.5 Hz, 1H), 6.96 (d, J=3.8 Hz, 1H),
2.19-2.16 (m, 2H), 1.93 (m, 4H), 1.81-1.78 (m, 2H).
Example 28
Synthesis of
6'-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4'-chlorospiro[cyclohexane-1-
,1'-isoindolin]-3'-one (Cpd. No. 28)
##STR00043##
[0292] Synthesis of
5'-bromo-7'-chloro-2'-[(4-methoxyphenyl)methyl]spiro[cyclohexane-1,3'-iso-
indoline]-1'-one (3)
[0293] To a solution of
5-bromo-7-chloro-2-[(4-methoxyphenyl)methyl]isoindolin-1-one (1,
2.0 g, 5.45 mmol) in tetrahydrofuran (25 mL) at room temperature
was added sodium hydride (654 mg, 27.27 mmol). The reaction was
stirred for 30 min and then 1,5-diiodopentane (2, 8835 mg, 27.27
mmol) was added to the reaction mixture. After stirring at room
temperature for 5 h, he reaction mass was quenched with a cold
solution of saturated ammonium chloride solution at 0.degree. C.
The residue was dissolved in ethyl acetate (100 mL) and the organic
layer was washed with water (2.times.20 mL) then with brine
solution (10 mL). The organic layers were separated and dried using
magnesium sulfate, filtered and concentrated. The crude was then
purified by flash column chromatography eluting with 10% ethyl
acetate in hexane. The desired fractions were concentrated to
dryness under vacuum to afford
5'-bromo-7'-chloro-2'-[(4-methoxyphenyl)
methyl]spiro[cyclohexane-1,3'-isoindoline]-1'-one (3) as a yellow
solid. Yield: 1.4 g, 60%. MS (ESI) m/z 436.44[M+1].sup.+; .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 8.06 (s, 1H), 7.84 (s, 1H),
7.23 (d, J=8.10 Hz, 2H), 6.87 (d, J=8.10 Hz, 2H), 4.64 (s, 2H),
3.72 (s, 3H), 1.98-1.90 (m, 3H), 1.78-1.71 (m, 5H), 1.49-1.40 (m,
2H).
Synthesis of
6'-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4'-chloro-2'-(4-methoxybenzy-
l)spiro[cyclohexane-1,1'-isoindolin]-3'-one (5)
[0294] The synthesis of intermediate 5 was carried out according to
the general protocol described above in Procedure C. brown coloured
solid. Yield: 0.08 g, 29%. MS (ESI) m/z 488.59 [M+1.sup.+; LCMS:
89%
Synthesis of
6'-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4'-chlorospiro[cyclohexane-1-
,1'-isoindolin]-3'-one (Cpd. No. 28)
[0295] The synthesis of compound 28 was carried out according to
the general protocol described above in Procedure F. Brown solid;
Yield: 0.025 g, 55%; MS (ESI) m/z 368.33[M+1].sup.+; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.32 (s, 1H), 8.25 (s, 1H), 8.20
(s, 1H), 8.08 (s, 1H) 7.71 (d, J=3.72 Hz, 1H), 7.24 (s, 2H), 6.84
(d, J=3.72 Hz, 1H), 2.01 (s, 2H), 1.70 (m, 5H), 1.43-1.40 (m,
3H).
Example 29
Synthesis of
4'-chloro-6'-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclohexane-1,1'-isoi-
ndolin]-3'-one (Cpd. No. 29)
##STR00044##
[0296] Synthesis of
4'-chloro-2'-(4-methoxybenzyl)-6'-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[-
cyclohexane-1,1'-isoindolin]-3'-one (3)
[0297] The synthesis of intermediate 3 was carried out according to
the general protocol described above in Procedure C. Brown solid;
Yield: 0.18 g, 64%; MS (ESI) m/z 473.4[M+1].sup.+; .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.16 (s, 1H), 8.93 (s, 1H), 8.27 (s,
1H), 8.16 (s, 1H), 7.34-7.26 (m, 2H), 6.94 (s, 1H), 6.88-6.86 (m,
2H), 6.57 (bs, 1H), 4.69 (s, 2H), 3.71 (s, 3H), 1.98-1.93 (m, 4H),
1.82-1.75 (m, 3H), 1.41-1.39 (s, 3H).
Synthesis of
4'-chloro-6'-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclohexane-1,1'-isoi-
ndolin]-3'-one (Cpd. No. 29)
[0298] The synthesis of compound 29 was carried out according to
the general protocol described above in Procedure F. Brown solid;
Yield: 0.050 g, 45%; MS (ESI) m/z 352.87[M+1].sup.+; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.39 (s, 1H), 9.16 (s, 1H), 8.96
(s, 1H), 8.25-8.23 (m, 2H), 8.16 (d, J=1.6 Hz, 1H), 6.96 (d, J=3.72
Hz, 1H), 2.05-1.97 (m, 2H), 1.70 (m, 5H), 1.45-1.43 (m, 3H).
Example 30
Synthesis of
6'-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4'-chlorospiro[cyclopentane--
1,1'-isoindolin]-3'-one (Cpd. No. 30)
##STR00045##
[0299] Synthesis of
6'-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4'-chloro-2'-(4-methoxybenzy-
l)spiro[cyclopentane-1,1'-isoindolin]-3'-one (3)
[0300] The synthesis of intermediate 3 was carried out according to
the general protocol described above in Procedure C. Brown solid;
Yield: 0.20 g, crude; MS (ESI) m/z 474.35[M+1].sup.+.
Synthesis of
6'-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4'-chlorospiro[cyclopentane--
1,1'-isoindolin]-3'-one (Cpd. No. 30)
[0301] The synthesis of compound 30 was carried out according to
the general protocol described above in Procedure F. Brown solid;
Yield: 0.020 g, 55%; MS (ESI) m/z 358.28[M+1].sup.+; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.08 (s, 1H), 8.23 (s, 1H), 8.19
(s, 1H), 8.05 (s, 1H) 7.79 (d, J=3.52 Hz, 1H), 7.24 (s, 2H), 6.84
(d, J=3.6 Hz, 1H), 2.17-1.77 (m, 8H).
Example 31
Synthesis of 6-(4-amino-7H-pyrrolo
[2,3-d]pyrimidin-7-yl)-1',4-dimethylspiro[isoindoline-1,4'-piperidin]-3-o-
ne (Cpd. No. 31)
##STR00046##
[0302] Synthesis of 6-(4-amino-7H-pyrrolo [2, 3-d]
pyrimidin-7-yl)-2-(4-methoxybenzyl)-1',4-dimethylspiro[isoindoline-1,4'-p-
iperidin]-3-one (3)
[0303] The synthesis of intermediate 3 was carried out according to
the general protocol described above in Procedure C. Light brown
solid; Yield: 0.25 g, crude; MS (ESI) m/z 483[M+1].sup.+.
Synthesis of 6-(4-amino-7H-pyrrolo
[2,3-d]pyrimidin-7-yl)-1',4-dimethylspiro[isoindoline-1,4'-piperidin]-3-o-
ne (Cpd. No. 31)
[0304] The synthesis of compound 31 was carried out according to
the general protocol described above in Procedure F. Off white
solid; Yield: 30 mg, 16%; MS (ESI) m/z 363.19[M+1].sup.+; .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 9.14 (s, 1H), 8.16 (s, 1H),
7.93 (s, 1H), 7.78 (s, 1H), 7.69 (d, J=3.7 Hz, 1H), 7.19 (s, 2H),
6.80 (d, J=3.7 Hz, 1H), 2.75-2.85 (m, 2H), 2.65 (s, 3H), 2.40-2.17
(m, 7H), 1.41 (m, 2H).
Example 32
Synthesis of
2-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin--
5-one (Cpd. No. 32)
##STR00047##
[0305] Synthesis of
6-(4-methoxybenzyl)-2-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)-6,7-dihydro-5H-py-
rrolo[3,4-b]pyridin-5-one (3)
[0306] The synthesis of intermediate 3 was carried out according to
the general protocol described above in Procedure C. Off white
solid; Yield: 0.060 g, 16%; MS (ESI) m/z 372.11[M+1].sup.+.
Synthesis of
2-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]
pyridin-5-one (Cpd. No. 32)
[0307] The synthesis of compound 32 was carried out according to
the general protocol described above in Procedure F. Off white
solid; Yield: 0.040 g, 55%; MS (ESI) m/z 252.07[M+1].sup.+; .sup.1H
NMR (400 MHz, DMSO-d.sub.6 with TFA-d) .delta. 9.54 (s, 1H), 9.36
(s, 1H), 8.74-8.72 (m, 2H), 8.34 (d, J=8.4 Hz, 1H), 7.22 (d, J=4.0
Hz, 1H), 4.45 (s, 2H).
Example 33
Synthesis of
6-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1'-(2,2-difluoroethyl)-4-meth-
ylspiro[isoindoline-1,4'-piperidin]-3-one (Cpd. No. 33)
##STR00048##
[0308] Synthesis of 6-(4-amino-7H-pyrrolo
[2,3-d]pyrimidin-7-yl)-1'-(2,2-difluoroethyl)-2-(4-methoxybenzyl)-4-methy-
lspiro[isoindoline-1,4'-piperidin]-3-one (3)
[0309] The synthesis of intermediate 3 was carried out according to
the general protocol described above in Procedure C. Brown solid;
Yield: 0.35 g, crude; MS (ESI) m/z 532.24[M+1].sup.+.
Synthesis of
6-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1'-(2,2-difluoroethyl)-4-meth-
ylspiro[isoindoline-1,4'-piperidin]-3-one (Cpd. No. 33)
[0310] The synthesis of compound 33 was carried out according to
the general protocol described above in Procedure F. White solid;
Yield: 7 mg, 3%, MS (ESI) m/z 412.18[M+1].sup.+; .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.162 (s, 1H), 8.17 (s, 1H), 7.95 (s,
1H), 7.83 (s, 1H), 7.70 (d, J=3.72 Hz, 1H), 7.16 (s, 2H), 6.80 (d,
J=3.64 Hz, 1H), 6.31-6.03 (tt, J=55.8, 3.46 Hz, 1H) 2.92-2.87 (m,
2H), 2.86-2.77 (m, 2H), 2.71-2.63 (m, 2H), 2.66 (s, 3H), 2.23-2.18
(m, 2H), 1.40-1.33 (m, 2H).
Example 34
Synthesis of
2'-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4'-methylspiro[cyclohexane-1-
,7'-pyrrolo[3,4-b]pyridin]-5'(6'H)-one (Cpd. No. 34)
##STR00049##
[0312] Procedure G: To a solution of
7H-pyrrolo[2,3-d]pyrimidin-4-amine (1, 0.32 g, 2.39 mmol) and
2'-chloro-4'-methylspiro[cyclohexane-1,7'-pyrrolo[3,4-b]pyridin]-5'(6'H)--
one (2, 0.6 g, 2.39 mmol) in 1,4-dioxane (15 mL) was added cesium
carbonate (2.33 g, 7.17 mmol). The reaction mixture was purged with
argon for 5 min. and then XanthPhos (69 mg, 0.11 mmol), XPhos (57
mg, 0.11 mmol), tris(dibenzylideneacetone)dipalladium(0) (109 mg,
0.11 mmol) and palladium acetate (27 mg, 0.11 mmol) were added and
the reaction mixture purged for an additional 5 min. The purged
reaction mixture was stirred at 100.degree. C. for 4 h. After TLC
showed completion, the reaction mixture was filtered through a bed
of celite and the resulting filtrate was concentrated. The crude
product was purified by preparative HPLC. The desired fractions
were concentrated to dryness under vacuum to afford
2'-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4'-methylspiro[cyclohexane-1-
,7'-pyrrolo[3,4-b]pyridin]-5'(6'H)-one as a yellow solid. Yield:
0.095 g, 11%; MS (ESI) m/z 348.4[M+1].sup.+; .sup.1H NMR: (400 MHz,
DMSO-d.sub.6) .delta. 12.36-12.28 (bs, 1H), 11.04-10.90 (bs, 1H),
9.12 (s, 1H), 8.64 (s, 1H), 7.10 (bs, 1H), 7.46 (s, 1H), 7.14 (s,
1H), 2.62 (s, 3H), 2.11-2.06 (m, 2H), 1.72-1.68 (m, 5H), 1.41-1.38
(m, 3H).
Example 35
Synthesis of
7-(3',4'-dimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-yl)-7H-py-
rrolo[2,3-d]pyrimidin-4-amine (Cpd. No. 35)
##STR00050##
[0313] Synthesis of
6'-methoxy-4'-methyl-3'-methylene-2',3'-dihydrospiro[cyclohexane-1,1'-ind-
ene] (2)
[0314] To a suspension of potassium tert-butoxide (1.75 g, 15.60
mmol) in tetrahydrofuran (20 mL) is added
methyltriphenylphosphonium bromide (5.46 g, 15.28 mmol). The
reaction is stirred at room temperature for 1 h and then cooled to
0.degree. C. A solution of
6'-methoxy-4'-methylspiro[cyclohexane-1,1'-inden]-3'(2'H)-one (1,
3.15 g, 12.89 mmol) in tetrahydrofuran (10 mL) is added. The
mixture is stirred at room temperature for 16 h, poured into water
and extracted with ethyl acetate. The organic phase is dried over
magnesium sulfate, filtered and concentrated. Purification via
column chromatography affords
6'-methoxy-4'-methyl-3'-methylene-2',3'-dihydrospiro[cyclohexane-1,1'-ind-
ene] (2).
Synthesis of
6'-methoxy-3',4'-dimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-indene]
(3)
[0315] To a solution of
6'-methoxy-4'-methyl-3'-methylene-2',3'-dihydrospiro
[cyclohexane-1,1'-indene] (2, 1.00 g, 4.13 mmol) in ethanol (20 mL)
is added 10% palladium on carbon (100 mg). The reaction is purged
with hydrogen and stirred at room temperature overnight. The
mixture is filtered through a pad of celite, concentrated and
purified via column chromatography to afford
6'-methoxy-3',4'-dimethyl-2',3'-dihydrospiro
[cyclohexane-1,1'-indene] (3).
Synthesis of
3',4'-dimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-ol
(4)
[0316] To a solution of
6'-methoxy-3',4'-dimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-indene]
(3, 1.00 g, 4.09 mmol) in dichloromethane (20 mL) at -78.degree. C.
is added slowly boron tribromide (0.79 mL, 8.18 mmol). The reaction
is allowed to stir at room temperature for 16 h. After completion,
the reaction mixture is quenched with saturated aqueous sodium
bicarbonate solution to adjust to pH 8. The mixture is extracted
with dichloromethane (2.times.30 mL). The combined organics is
dried over sodium sulfate, filtered and concentrated. The crude is
then purified via column chromatography to afford
3',4'-dimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-ol
(4).
Synthesis of
3',4'-dimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-yl
trifluoromethanesulfonate (5)
[0317] To a solution of 3',4'-dimethyl-2',3'-dihydrospiro
[cyclohexane-1,1'-inden]-6'-ol (4, 1.00 g, 4.34 mmol) in
dichloromethane (15 mL) at -30.degree. C., diisopropylethylamine
(1.28 mL, 7.38 mmol) is added followed by the slow addition of
triflic anhydride (0.80 mL, 4.77 mmol). The reaction is allowed to
stir at room temperature for 1 h. After completion, the reaction
mixture is basified by saturated aqueous sodium bicarbonate
solution to pH 8. The mixture is extracted with dichloromethane
(2.times.10 mL). The combined organics is dried over sodium
sulfate, filtered and concentrated to dryness under vacuum. The
crude is then purified via column chromatography to afford
3',4'-dimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-yl
trifluoromethanesulfonate (5).
Synthesis of
7-(3',4'-dimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-yl)-7H-py-
rrolo[2,3-d]pyrimidin-4-amine (Cpd. No. 35)
[0318] The synthesis of compound 35 is carried out as described
above using the general protocol of Procedure G.
Example 36
Synthesis of
7-(3',3',4'-trimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-yl)-7-
H-pyrrolo[2,3-d]pyrimidin-4-amine (Cpd. No. 36)
##STR00051## ##STR00052##
[0319] Synthesis of methyl
2-(4-methoxy-2-methylphenyl)-2-methylpropanoate (2)
[0320] To a solution of methyl 2-(4-methoxy-2-methylphenyl)acetate
(1, 1.00 g, 5.15 mmol) in tetrahydrofuran (20 mL) at 0.degree. C.,
sodium hydride (0.31 g, 12.88 mmol) is added portion wise and the
reaction mixture is allowed to stir at room temperature for 30 min.
Iodomethane (0.96 mL, 15.45 mmol) is added and the reaction mixture
is allowed to stir at 70.degree. C. for 16 h. The reaction mixture
is quenched with water and is extracted in ethyl acetate. The
organic layer is separated, dried over sodium sulphate, filtered
and concentrated under reduced pressure. The residue is purified by
silica gel column chromatography to afford methyl
2-(4-methoxy-2-methylphenyl)-2-methylpropanoate (2).
Synthesis of 2-(4-methoxy-2-methylphenyl)-2-methylpropanoic acid
(3)
[0321] To a solution of methyl
2-(4-methoxy-2-methylphenyl)-2-methylpropanoate (2, 1.00 g, 4.50
mmol) in tetrahydrofuran (10 mL) and ethanol (10 mL) is added 1 M
lithium hydroxide aqueous solution (10 mL). The reaction is stirred
at room temperature overnight. The mixture is diluted with water
and extracted with ethyl acetate. The organic layer is washed with
brine, dried over sodium sulfate, filtered and concentrated. The
crude obtained is further purified via column chromatography to
obtain 2-(4-methoxy-2-methylphenyl)-2-methylpropanoic acid (3).
Synthesis of
1-diazo-3-(4-methoxy-2-methylphenyl)-3-methylbutan-2-one (4)
[0322] To a solution of
2-(4-methoxy-2-methylphenyl)-2-methylpropanoic acid (3, 1.00 g,
4.80 mmol) in dichloromethane (10 mL) at 0.degree. C. is added
oxalyl chloride (1 M in dichloromethane, 5.28 mL, 5.28 mmol)
followed by two drops of N,N-dimethylformamide. The reaction is
stirred at room temperature for 1 h. The mixture is concentrated
and dried under vacuum. The residue is dissolved in dichloromethane
(10 mL). To this solution at 0.degree. C. is purged with
diazomethane. The reaction is fitted with a calcium chloride drying
tube and allowed to stand at room temperature for 16 h. The mixture
is purged with nitrogen and concentrated. The residue is purified
via column chromatography to afford
1-diazo-3-(4-methoxy-2-methylphenyl)-3-methylbutan-2-one (4).
Synthesis of 5-methoxy-1,1,7-trimethyl-1,3-dihydro-2H-inden-2-one
(5)
[0323] To a solution of
1-diazo-3-(4-methoxy-2-methylphenyl)-3-methylbutan-2-one (4, 1.00
g, 4.30 mmol) in dichloromethane (10 mL) is added rhodium (II)
acetate dimer dihydrate (105 mg, 0.22 mmol). The reaction is
stirred at room temperature overnight. The mixture is filtered
through a pad of celite, concentrated and purified via column
chromatography to afford
5-methoxy-1,1,7-trimethyl-1,3-dihydro-2H-inden-2-one (5).
Synthesis of
6'-methoxy-3',3',4'-trimethylspiro[cyclohexane-1,1'-inden]-2'(3'H)-one
(7)
[0324] To a solution of
5-methoxy-1,1,7-trimethyl-1,3-dihydro-2H-inden-2-one (5, 1.00 g,
4.90 mmol) in tetrahydrofuran (20 mL) at 0.degree. C., sodium
hydride (0.29 g, 12.25 mmol) is added portion wise and the reaction
mixture is allowed to stir at room temperature for 30 min.
1,5-Dibromopentane (6, 1.13 g, 4.9 mmol) is added and the reaction
mixture is allowed to stir at 70.degree. C. for 16 h. The reaction
mixture is quenched with water and is extracted in ethyl acetate.
The organic layer is separated, dried over sodium sulphate,
filtered and concentrated under reduced pressure. The residue is
purified by silica gel column chromatography to afford
6'-methoxy-3',3',4'-trimethylspiro[cyclohexane-1,1'-inden]-2'(3'H)-one
(7).
Synthesis of
6'-methoxy-3',3',4'-trimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-indene]
(8)
[0325] To a solution of
6'-methoxy-3',3',4'-trimethylspiro[cyclohexane-1,1'-inden]-2'(311)-one
(7, 1.00 g, 3.67 mmol) in ethylene glycol (40 mL) is added
hydrazine hydrate solution (78-82%, 0.25 g, 4.04 mmol) followed by
potassium hydroxide (0.62 g, 11.01 mmol). The reaction is fitted
with a Dean-Stark trap and stirred at 120.degree. C. for 3 h to
distill off water and excess hydrazine. The reaction is then
stirred at reflux overnight. The mixture is cooled to room
temperature, diluted with water and extracted with ethyl acetate.
The combined organics is dried over magnesium sulfate, filtered and
concentrated. The crude is purified via column chromatography to
afford
6'-methoxy-3',3',4'-trimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-indene]
(8).
Synthesis of
3',3',4'-trimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-ol
(9)
[0326] To a solution of
6'-methoxy-3',3',4'-trimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-indene]
(8, 1.00 g, 3.87 mmol) in dichloromethane (20 mL) at -78.degree. C.
is added slowly boron tribromide (0.74 mL, 7.74 mmol). The reaction
is allowed to stir at room temperature for 16 h. After completion,
the reaction mixture is quenched with saturated aqueous sodium
bicarbonate solution to adjust to pH 8. The mixture is extracted
with dichloromethane (2.times.30 mL). The combined organics is
dried over sodium sulfate, filtered and concentrated. The crude is
then purified via column chromatography to afford
3',3',4'-trimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-ol
(9).
Synthesis of
3',3',4'-trimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-yl
trifluoromethanesulfonate (10)
[0327] To a solution of
3',3',4'-trimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-ol
(9, 1.00 g, 4.09 mmol) in dichloromethane (15 mL) at -30.degree.
C., diisopropylethylamine (1.21 mL, 6.95 mmol) is added followed by
the slow addition of triflic anhydride (0.76 mL, 4.50 mmol). The
reaction is allowed to stir at room temperature for 1 h. After
completion, the reaction mixture is basified by saturated aqueous
sodium bicarbonate solution to pH 8. The mixture is extracted with
dichloromethane (2.times.20 mL). The combined organics is dried
over sodium sulfate, filtered and concentrated to dryness under
vacuum. The crude is then purified via column chromatography to
afford
3',3',4'-trimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-yl
trifluoromethanesulfonate (10).
Synthesis of
7-(3',3',4'-trimethyl-2',3'-dihydrospiro[cyclohexane-1,1'-inden]-6'-yl)-7-
H-pyrrolo[2,3-d]pyrimidin-4-amine (Cpd. No. 36)
[0328] The synthesis of compound 36 is carried out as described
above using the general protocol of Procedure G.
Example 37
Synthesis of
7-(3',4'-dimethylspiro[cyclohexane-1,1'-isoindolin]-6'-yl)-7H-pyrrolo[2,3-
-d]pyrimidin-4-amine (Cpd. No. 37)
##STR00053## ##STR00054##
[0329] Synthesis of
6'-methoxy-4'-methylspiro[cyclohexane-1,1'-isoindoline] (2)
[0330] To a solution of
6'-methoxy-4'-methylspiro[cyclohexane-1,1'-isoindolin]-3'-one (1,
1.00 g, 4.08 mmol) in tetrahydrofuran (20 mL) is added dropwise
borane dimethyl sulfide complex (12.24 mL, 24.48 mmol, 2 M in
tetrahydrofuran). The reaction is stirred at 65.degree. C. for 7 h,
then stirred at room temperature overnight. 0.5 M hydrochloric acid
(8 mL) is added dropwise and the mixture is refluxed for 2 h. The
mixture is cooled to room temperature, basified with 1 M aqueous
sodium hydroxide solution to pH=8 and extracted with ethyl acetate.
The combined organics is dried over magnesium sulfate, filtered and
concentrated. The crude is purified via column chromatography to
afford methyl
6'-methoxy-4'-methylspiro[cyclohexane-1,1'-isoindoline] (2).
Synthesis of
N-tert-butyl-1-(6'-methoxy-4'-methylspiro[cyclohexane-1,1'-isoindolin]-2'-
-yl)methanimine (4)
[0331] To a solution of methyl
6'-methoxy-4'-methylspiro[cyclohexane-1,1'-isoindoline] (2, 1.00 g,
4.32 mmol) in toluene (20 mL) is added ammonium sulfate (1.14 g,
8.64 mmol) followed by N'-tert-butyl-N,N-dimethylformimidamide (3,
0.83 g, 6.48 mmol). The reaction is refluxed overnight. The mixture
is cooled to room temperature, filtered and concentrated. The crude
is purified via column chromatography to afford
N-tert-butyl-1-(6'-methoxy-4'-methylspiro[cyclohexane-1,1'-isoindolin]-2'-
-yl)methanimine (4).
Synthesis of
N-tert-butyl-1-(6'-methoxy-3',4'-dimethylspiro[cyclohexane-1,1'-isoindoli-
n]-2'-yl)methanimine (5)
[0332] To a solution of
N-tert-butyl-1-(6'-methoxy-4'-methylspiro[cyclohexane-1,1'-isoindolin]-2'-
-yl)methanimine (4, 1.00 g, 3.18 mmol) in tetrahydrofuran (20 mL)
at -78.degree. C. is added n-butyl lithium (1.6 M in hexanes, 2.19
mL, 3.50 mmol) dropwise and the reaction is stirred for 30 min.
Iodomethane (0.30 mL, 4.77 mmol) is added and the reaction is
warmed to room temperature and stirred for 1 h. The reaction
mixture is quenched with water and extracted with ethyl acetate.
The combined organics is dried over magnesium sulphate, filtered
and concentrated. The residue is purified via column chromatography
to
N-tert-butyl-1-(6'-methoxy-3',4'-dimethylspiro[cyclohexane-1,1'-isoindoli-
n]-2'-yl)methanimine (5).
Synthesis of 3',4'-dimethylspiro[cyclohexane-1,1'-isoindolin]-6'-ol
(6)
[0333] To a solution of
N-tert-butyl-1-(6'-methoxy-3',4'-dimethylspiro[cyclohexane-1,1'-isoindoli-
n]-2'-yl)methanimine (5, 1.00 g, 3.04 mmol) in dichloromethane (20
mL) at -78.degree. C. is added slowly boron tribromide (0.59 mL,
6.08 mmol). The reaction is stirred at room temperature for 16 h.
After completion, the reaction mixture is quenched with saturated
aqueous sodium bicarbonate solution to adjust to pH 8. The mixture
is extracted with dichloromethane (2.times.30 mL). The combined
organics is dried over sodium sulfate, filtered and concentrated.
The crude is then purified via column chromatography to afford
3',4'-dimethylspiro[cyclohexane-1,1'-isoindolin]-6'-ol (6).
Synthesis of tert-butyl
6'-hydroxy-3',4'-dimethylspiro[cyclohexane-1,1'-isoindoline]-2'-carboxyla-
te (7)
[0334] To a solution of
3',4'-dimethylspiro[cyclohexane-1,1'-isoindolin]-6'-ol (6, 1.00 g,
4.32 mmol) and di-tert-butyl dicarbonate (1.19 mL, 5.18 mmol) in
tetrahydrofuran (20 mL) is added a solution of potassium carbonate
(1.49 g, 10.80 mmol) in water (20 mL). The reaction is stirred at
room temperature overnight. The mixture is diluted with brine and
extracted with ethyl acetate. The combined organics is dried over
magnesium sulfate, filtered and concentrated. The crude is then
purified via column chromatography to afford tert-butyl
6'-hydroxy-3',4'-dimethylspiro[cyclohexane-1,1'-isoindoline]-2'-carboxyla-
te (7).
Synthesis of tert-butyl
3',4'-dimethyl-6'-(((trifluoromethyl)sulfonyl)oxy)spiro[cyclohexane-1,1'--
isoindoline]-2'-carboxylate (8)
[0335] To a solution of tert-butyl
6'-hydroxy-3',4'-dimethylspiro[cyclohexane-1,1'-isoindoline]-2'-carboxyla-
te (7, 1.00 g, 3.02 mmol) in dichloromethane (15 mL) at -30.degree.
C., diisopropylethylamine (0.89 mL, 5.13 mmol) is added followed by
the slow addition of triflic anhydride (0.56 mL, 3.32 mmol). The
reaction is allowed to stir at room temperature for 1 h. After
completion, the reaction mixture is basified by saturated aqueous
sodium bicarbonate solution to pH 8. The mixture is extracted with
dichloromethane (2.times.20 mL). The combined organics is dried
over sodium sulfate, filtered and concentrated to dryness under
vacuum. The crude is then purified via column chromatography to
afford tert-butyl
3',4'-dimethyl-6'-(((trifluoromethyl)sulfonyl)oxy)spiro[cyclohexane-1,1'--
isoindoline]-2'-carboxylate (8).
Synthesis of tert-butyl
6'-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3',4'-dimethylspiro[cyclohex-
ane-1,1'-isoindoline]-2'-carboxylate (10)
[0336] The synthesis of intermediate 10 is carried out as described
above using the general protocol of Procedure G.
Synthesis of
7-(3',4'-dimethylspiro[cyclohexane-1,1'-isoindolin]-6'-yl)-7H-pyrrolo[2,3-
-d]pyrimidin-4-amine (Cpd. No. 37)
[0337] The synthesis of intermediate 37 is carried out as described
above using the general protocol of Procedure C.
Example 38
Synthesis of
7-(3',3',4'-trimethylspiro[cyclohexane-1,1'-isoindolin]-6'-yl)-7H-pyrrolo-
[2,3-d]pyrimidin-4-amine (Cpd. No. 38)
##STR00055## ##STR00056##
[0338] Synthesis of
N-tert-butyl-1-(6'-methoxy-3',3',4'-trimethylspiro[cyclohexane-1,1'-isoin-
dolin]-2'-yl)methanimine (2)
[0339] To a solution of
N-tert-butyl-1-(6'-methoxy-3',4'-dimethylspiro[cyclohexane-1,1'-isoindoli-
n]-2'-yl)methanimine (1, 1.00 g, 3.04 mmol) in tetrahydrofuran (20
mL) at -78.degree. C. is added n-butyl lithium (1.6 M in hexanes,
2.09 mL, 3.34 mmol) dropwise and the reaction is stirred for 30
min. Iodomethane (0.28 mL, 4.56 mmol) is added and the reaction is
warmed to room temperature and stirred for 1 h. The reaction
mixture is quenched with water and extracted with ethyl acetate.
The combined organics is dried over magnesium sulphate, filtered
and concentrated. The residue is purified via column chromatography
to
N-tert-butyl-1-(6'-methoxy-3',4'-dimethylspiro[cyclohexane-1,1'-isoindoli-
n]-2'-yl)methanimine (2).
Synthesis of
3',3',4'-trimethylspiro[cyclohexane-1,1'-isoindolin]-6'-ol (3)
[0340] To a solution of
N-tert-butyl-1-(6'-methoxy-3',4'-dimethylspiro[cyclohexane-1,1'-isoindoli-
n]-2'-yl)methanimine (2, 1.00 g, 2.92 mmol) in dichloromethane (20
mL) at -78.degree. C. is added slowly boron tribromide (0.56 mL,
5.84 mmol). The reaction is stirred at room temperature for 16 h.
After completion, the reaction mixture is quenched with saturated
aqueous sodium bicarbonate solution to adjust to pH 8. The mixture
is extracted with dichloromethane (2.times.30 mL). The combined
organics is dried over sodium sulfate, filtered and concentrated.
The crude is then purified via column chromatography to afford
3',3',4'-trimethylspiro[cyclohexane-1,1'-isoindolin]-6'-ol (3).
Synthesis of tert-butyl
6'-hydroxy-3',3',4'-trimethylspiro[cyclohexane-1,1'-isoindoline]-2'-carbo-
xylate (4)
[0341] To a solution of
3',3',4'-trimethylspiro[cyclohexane-1,1'-isoindolin]-6'-ol (3, 1.00
g, 4.08 mmol) and di-tert-butyl dicarbonate (1.12 mL, 4.90 mmol) in
tetrahydrofuran (20 mL) is added a solution of potassium carbonate
(1.41 g, 10.20 mmol) in water (20 mL). The reaction is stirred at
room temperature overnight. The mixture is diluted with brine and
extracted with ethyl acetate. The combined organics is dried over
magnesium sulfate, filtered and concentrated. The crude is then
purified via column chromatography to afford tert-butyl
6'-hydroxy-3',3',4'-trimethylspiro[cyclohexane-1,1'-isoindoline]-2'-carbo-
xylate (4).
Synthesis of tert-butyl
3',3',4'-trimethyl-6'-(((trifluoromethyl)sulfonyl)oxy)spiro[cyclohexane-1-
,1'-isoindoline]-2'-carboxylate (5)
[0342] To a solution of tert-butyl
6'-hydroxy-3',3',4'-trimethylspiro[cyclohexane-1,1'-isoindoline]-2'-carbo-
xylate (4, 1.00 g, 2.89 mmol) in dichloromethane (15 mL) at
-30.degree. C., diisopropylethylamine (0.86 mL, 4.91 mmol) is added
followed by the slow addition of triflic anhydride (0.54 mL, 3.18
mmol). The reaction is allowed to stir at room temperature for 1 h.
After completion, the reaction mixture is basified by saturated
aqueous sodium bicarbonate solution to pH 8. The mixture is
extracted with dichloromethane (2.times.20 mL). The combined
organics is dried over sodium sulfate, filtered and concentrated to
dryness under vacuum. The crude is then purified via column
chromatography to afford tert-butyl
3',3',4'-trimethyl-6'-(((trifluoromethyl)sulfonyl)oxy)spiro[cyclohexane-1-
,1'-isoindoline]-2'-carboxylate (5).
Synthesis of tert-butyl
6'-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3',3',4'-trimethylspiro[cycl-
ohexane-1,1'-isoindoline]-2'-carboxylate (7)
[0343] The synthesis of intermediate 7 is carried out as described
above using the general protocol of Procedure G.
Synthesis of
7-(3',3',4'-trimethylspiro[cyclohexane-1,1'-isoindolin]-6'-yl)-7H-pyrrolo-
[2,3-d]pyrimidin-4-amine (Cpd. No. 38)
[0344] The synthesis of compound 38 is carried out as described
above using the general protocol of Procedure C.
Example 39
Synthesis of
7-(4'-methyl-2'H-dispiro[cyclohexane-1,1'-indene-3',1''-cyclopropan]-6'-y-
l)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (Cpd. No. 39)
##STR00057##
[0345] Synthesis of
6'-methoxy-4'-methyl-2'H-dispiro[cyclohexane-1,1'-indene-3',1''-cycloprop-
ane] (2)
[0346] A 1.1 M toluene solution of diethyl zinc (15.02 mL, 16.52
mmol) is added to a reaction vessel containing dichloromethane (20
mL) and cooled to 0.degree. C. Trifluoroacetic acid (1.26 mL, 16.52
mmol) is added to the resulting solution and the reaction is
stirred at 0.degree. C. for 15 min. To the cooled solution is added
diiodomethane (1.33 mL, 16.52 mmol), and the reaction is stirred
for an additional 15 min at 0.degree. C. Then, a solution of
6'-methoxy-4'-methyl-3'-methylene-2',3'-dihydrospiro[cyclohexane-1,1'-ind-
ene](1, 1.00 g, 4.13 mmol) in dichloromethane (10 mL) is added. The
reaction is maintained 0.degree. C. for another 15 min, then
allowed to gradually warm to room temperature. Upon completion, the
reaction is quenched with a saturated aqueous ammonium chloride
solution (40 mL) and diluted with dichloromethane (40 mL). The
combined organics is washed with brine (40 mL), dried over
magnesium sulfate, filtered and concentrated. The crude is purified
via flash chromatography to afford
6'-methoxy-4'-methyl-2'H-dispiro[cyclohexane-1,1'-indene-3',1''-cycloprop-
ane] (2).
Synthesis of
4'-methyl-2'H-dispiro[cyclohexane-1,1'-indene-3',1''-cyclopropan]-6'-ol
(3)
[0347] To a solution of
6'-methoxy-4'-methyl-2'H-dispiro[cyclohexane-1,1'-indene-3',1''-cycloprop-
ane] (2, 0.75 g, 2.92 mmol) in dichloromethane (20 mL) at
-78.degree. C. is added slowly boron tribromide (0.56 mL, 5.84
mmol). The reaction is stirred at room temperature for 16 h. After
completion, the reaction mixture is quenched with saturated aqueous
sodium bicarbonate solution to adjust to pH 8. The mixture is
extracted with dichloromethane (2.times.30 mL). The combined
organics is dried over sodium sulfate, filtered and concentrated.
The crude is then purified via column chromatography to afford
4'-methyl-2'H-dispiro[cyclohexane-1,1'-indene-3',1''-cyclopropan]--
6'-ol (3).
Synthesis of
4'-methyl-2'H-dispiro[cyclohexane-1,1'-indene-3',1''-cyclopropan]-6'-yl
trifluoromethanesulfonate (4)
[0348] To a solution of
4'-methyl-2'H-dispiro[cyclohexane-1,1'-indene-3',1''-cyclopropan]-6'-ol
(3, 0.70 g, 2.89 mmol) in dichloromethane (15 mL) at -30.degree.
C., diisopropylethylamine (0.86 mL, 4.91 mmol) is added followed by
the slow addition of triflic anhydride (0.54 mL, 3.18 mmol). The
reaction is allowed to stir at room temperature for 1 h. After
completion, the reaction mixture is basified by saturated aqueous
sodium bicarbonate solution to pH 8. The mixture is extracted with
dichloromethane (2.times.20 mL). The combined organics is dried
over sodium sulfate, filtered and concentrated to dryness under
vacuum. The crude is then purified via column chromatography to
afford
4'-methyl-2'H-dispiro[cyclohexane-1,1'-indene-3',1''-cyclopropan]-6'-yl
trifluoromethanesulfonate (4).
Synthesis of
7-(4'-methyl-2'H-dispiro[cyclohexane-1,1'-indene-3',1''-cyclopropan]-6'-y-
l)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (Cpd. No. 39)
[0349] The synthesis of compound 39 is carried out as described
above using the general protocol of Procedure G.
Example 40: MNK Biochemical Enzymatic Assay
[0350] Compounds are screened for MNK inhibition using the ADP-Glo
kinase assay kit (Promega, catalogue No. V9101). All kinase
reactions are performed in Reaction Buffer E (15 mM HEPES pH7.4, 20
mM NaCl, 1 mM EGTA, 10 mM MgCl.sub.2, 0.1 mg/ml BGG, and 0.02%
Tween-20). Final MNK1 reactions contained 10 nM recombinant MNK1
(Life Technologies, PR9138A), 100 .mu.M MNK substrate peptide
Ac-TATKSGSTTKNR-NH2 (American Peptide Company), 300 .mu.M ATP, and
varying concentrations of the inhibitory compound of interest.
Final MNK2 reactions contained 3 nM recombinant MNK2 (Life
Technologies, PV5607), 50 .mu.M MNK substrate peptide
Ac-TATKSGSTTKNR-NH2 (American Peptide Company), 10 .mu.M ATP, and
varying concentrations of the inhibitory compound of interest.
Final DMSO concentration in each reaction is 1%.
[0351] Kinase reactions are carried out in 96-well half-area white
flat-bottom polystyrene plates in a final volume of 25 .mu.l.
MNK1/2 enzymes are pre-incubated with compound and peptide
substrate for 5 minutes prior to the addition of ATP. After the
addition of ATP, kinase reactions are incubated at room temperature
for 40 minutes. Reactions are subsequently stopped by the addition
of 25 .mu.l of ADP-Glo Reagent and incubating for an additional 40
minutes. The final luminescent signal used for kinase activity
readout is produced by the addition of 45 .mu.l of Kinase Detection
Reagent (ADP-Glo kit, Promega) and incubating for 40 minutes. The
luminescent signal is detected using a Victor 2 multilabel counter
(Perkin Elmer) and the concentration of compound necessary to
achieve inhibition of enzyme activity by 50% (IC.sub.50) is
calculated using signals from an 8-point compound dilution
series.
[0352] The results of these assays are set forth in Table 1 below.
To this end, IC.sub.50 values of less than 0.01 .mu.M are labeled
as "+++", from 0.01 to 0.1 .mu.M are labeled as "++", and greater
than 0.1 to 10.0 .mu.M are labeled as "+" (NA means "not
available").
TABLE-US-00001 TABLE 1 MNK Biochemical Enzymatic Assay (IC.sub.50)
IC.sub.50 Cpd. No. Mnk1 Mnk2 1 + + 2 NA + 3 NA + 4 NA + 5 NA + 6 NA
+ 7 NA + 8 ++ ++ 9 NA - 10 NA - 11 NA + 12 NA + 13 NA + 14 NA + 15
NA - 16 NA + 17 +++ ++ 18 +++ +++ 19 NA + 20 NA + 21 NA + 22 ++ ++
23 NA + 24 NA + 25 NA + 26 NA + 27 NA +++ 28 NA +++ 29 NA ++ 30 NA
+++ 31 NA NA 32 NA NA 33 NA NA 34 NA NA
Example 41: peIF4E Signaling Cellular Assay
[0353] Phosphorylated eIF4E is assayed using the CisBio peIF4E
HTRF.RTM. assay kit (CisBio, catalogue No. 64EF4PEG). Cells are
plated in 96-well tissue-culture treated plate in appropriate
growth medium (90 .mu.L). Compounds (10.times.) are diluted using
3-fold serial dilutions in cell culture medium and added to cells.
Plates are incubated for 2 hrs at 37.degree. C. The cell
supernatant is carefully removed either by aspirating supernatant
or by flicking the plate. Immediately 50 .mu.L of supplemented
lysis buffer (1.times.) is added and incubated for at least 30
minutes at room temperature under shaking. After homogenization by
pipeting up and down, 16 .mu.L of cell lysate is transferred from
the 96-well cell-culture plate to a 384-well small volume white
plate. 4 .mu.L of premixed antibody solutions (vol/vol) is prepared
in the detection buffer and added. The plate is covered with a
plate sealer and incubated overnight at room temperature. The
fluorescence emissions at two different wavelengths are read (665
nm and 620 nm) on a Wallac Victor2. Emission ratios are converted
into percent inhibitions and imported into GraphPad Prism software.
The concentration of compound necessary to achieve inhibition of
enzyme activity by 50% (IC.sub.50) is calculated using
concentrations ranging from 20 .mu.M to 0.1 nM (12-point curve).
IC.sub.50 values are determined using a nonlinear regression model
available in GraphPad Prism 5.
[0354] The results of these assays are set forth in Table 2 below.
To this end, IC.sub.50 values of less than 0.05 .mu.M are labeled
as "+++", from 0.05 to 1.0 .mu.M are labeled as "++", greater than
1.0 to 100 .mu.M are labeled as "+", and NA means "not
available".
TABLE-US-00002 TABLE 2 peIF4E Signaling Cellular Assay (IC.sub.50)
Cpd. No. IC.sub.50 1 + 2 NA 3 + 4 NA 5 NA 6 + 7 NA 8 + 9 NA 10 NA
11 + 12 NA 13 NA 14 NA 15 NA 16 NA 17 ++ 18 ++ 19 NA 20 NA 21 NA 22
+ 23 NA 24 NA 25 NA 26 NA 27 ++ 28 +++ 29 +++ 30 +++ 31 ++ 32 ++ 33
++ 34 +++
[0355] The various embodiments described above can be combined to
provide further embodiments. All of the U.S. patents, U.S. patent
application publications, U.S. patent applications, foreign
patents, foreign patent applications and non-patent publications
referred to in this specification and/or listed in the Application
Data Sheet are incorporated herein by reference, in their entirety.
Aspects of the embodiments can be modified, if necessary to employ
concepts of the various patents, applications and publications to
provide yet further embodiments.
[0356] These and other changes can be made to the embodiments in
light of the above-detailed description. In general, in the
following claims, the terms used should not be construed to limit
the claims to the specific embodiments disclosed in the
specification and the claims, but should be construed to include
all possible embodiments along with the full scope of equivalents
to which such claims are entitled. Accordingly, the claims are not
limited by the disclosure.
* * * * *