U.S. patent number 10,385,018 [Application Number 14/438,410] was granted by the patent office on 2019-08-20 for 4-carboxamido-isoindolinone derivatives as selective parp-1 inhibitors.
This patent grant is currently assigned to NERVIANO MEDICAL SCIENCES S.R.L.. The grantee listed for this patent is NERVIANO MEDICAL SCIENCES S.r.l. Invention is credited to Mikhail Yurievitch Krasavin, Paolo Orsini, Gianluca Mariano Enrico Papeo, Alessandra Scolaro.
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United States Patent |
10,385,018 |
Papeo , et al. |
August 20, 2019 |
4-carboxamido-isoindolinone derivatives as selective PARP-1
inhibitors
Abstract
There are provided substituted 4-carboxamido-isoindolinone
derivatives which selectively inhibit the activity of poly
(ADP-ribose) polymerase PARP-1 with respect to poly (ADP-ribose)
polymerase PARP-2. The compounds of this invention are therefore
useful in treating diseases such as cancer, cardiovascular
diseases, central nervous system injury and different forms of
inflammation. The present invention also provides methods for
preparing these compounds, pharmaceutical compositions comprising
these compounds, and methods of treating diseases utilizing
pharmaceutical compositions comprising these compounds.
Inventors: |
Papeo; Gianluca Mariano Enrico
(Cernusco Lombardone, IT), Krasavin; Mikhail
Yurievitch (Nathan, AU), Orsini; Paolo (Legnano,
IT), Scolaro; Alessandra (Bresso, IT) |
Applicant: |
Name |
City |
State |
Country |
Type |
NERVIANO MEDICAL SCIENCES S.r.l |
Nerviano (MI) |
N/A |
IT |
|
|
Assignee: |
NERVIANO MEDICAL SCIENCES
S.R.L. (Nerviano (MI), IT)
|
Family
ID: |
47115518 |
Appl.
No.: |
14/438,410 |
Filed: |
October 23, 2013 |
PCT
Filed: |
October 23, 2013 |
PCT No.: |
PCT/EP2013/072165 |
371(c)(1),(2),(4) Date: |
April 24, 2015 |
PCT
Pub. No.: |
WO2014/064149 |
PCT
Pub. Date: |
May 01, 2014 |
Prior Publication Data
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|
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Document
Identifier |
Publication Date |
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US 20150274662 A1 |
Oct 1, 2015 |
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Foreign Application Priority Data
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|
|
|
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Oct 26, 2012 [EP] |
|
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12190130 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K
31/496 (20130101); C07D 403/14 (20130101); A61K
31/4188 (20130101); A61K 31/4725 (20130101); C07D
405/04 (20130101); C07D 409/14 (20130101); C07D
413/06 (20130101); A61K 31/4035 (20130101); C07D
209/46 (20130101); C07D 401/14 (20130101); C07D
403/04 (20130101); C07D 413/14 (20130101); A61K
31/5377 (20130101); C07D 401/06 (20130101); A61P
35/00 (20180101); A61P 43/00 (20180101); A61P
9/00 (20180101); A61K 31/454 (20130101); A61P
29/00 (20180101); C07D 209/44 (20130101); C07D
403/02 (20130101); C07D 405/14 (20130101); C07D
407/14 (20130101); A61P 25/00 (20180101); A61K
31/4439 (20130101) |
Current International
Class: |
C07D
209/44 (20060101); C07D 413/14 (20060101); C07D
401/06 (20060101); C07D 403/02 (20060101); C07D
403/04 (20060101); C07D 403/14 (20060101); C07D
405/04 (20060101); A61K 31/5377 (20060101); C07D
409/14 (20060101); C07D 413/06 (20060101); C07D
209/46 (20060101); A61K 31/4035 (20060101); A61K
31/4188 (20060101); A61K 31/4439 (20060101); A61K
31/454 (20060101); A61K 31/4725 (20060101); A61K
31/496 (20060101); C07D 405/14 (20060101); C07D
401/14 (20060101); C07D 407/14 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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WO 2007/047646 |
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Apr 2007 |
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WO |
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WO 2010/133647 |
|
Nov 2010 |
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WO |
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WO 2011/006794 |
|
Jan 2011 |
|
WO |
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WO 2011/006803 |
|
Jan 2011 |
|
WO |
|
Other References
Gandhi "Discovery and SAR of substituted
3-oxoisoindoline-4-carboxamides as potent inhibitors of
poly(ADP-ribose) polymerase (PARP) for the treatment of cancer."
Bioorganic & Medicinal Chemistry Letters, 2010, 20(3),
1023-1026. cited by examiner .
Collins "18-Cycloalkyl Analogues of Enisoprost" J. Med. Chem. 1989,
32, 1001-1006. cited by examiner .
Bissery MC et al., "Experimental Antitumor Activity and
Pharmacokinetics of the Camptothecin Analog Irinotecan (CPT-11) in
Mice", Anti-Cancer Drugs 7:437-460 (1996). cited by applicant .
Gandhi V.B. et al., "Discovery and SAR of Substituted
3-Oxoisoindoline-4-Carboxamides as Potent Inhibitors of Poly
(ADP-Ribose) Polymerase (PARP) for the Treatment of Cancer",
Bioorganic & Medicinal Chemistry Letters 20:1023-1026 (2010).
cited by applicant .
Simeoni M. et al., "Predictive Pharmacokinetic-Pharmacodynamic
Modeling of Tumor Growth Kinetics in Xenograft Models After
Administration of Anticancer Agents", Cancer Research 64:1094-1101
(Feb. 1, 2004). cited by applicant .
International Search Report dated Mar. 7, 2014 issued in
PCT/EP2013/072165. cited by applicant.
|
Primary Examiner: O'Dell; David K
Attorney, Agent or Firm: Scully, Scott, Murphy &
Presser, P.C.
Claims
The invention claimed is:
1. A compound selected from the group consisting of:
2-(1-cyclohexyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxyl-
ic acid amide; and
2-(1-cyclohexyl-piperidin-4-yl)-6-fluoro-3-oxo-2,3-dihydro-1H-isoindole-4-
-carboxylic acid amide; or a pharmaceutically acceptable salt
thereof.
2. An in vitro method for selectively inhibiting PARP-1 protein
activity, which comprises contacting the said protein with an
effective amount of a compound of claim 1.
3. A pharmaceutical composition comprising a therapeutically
effective amount of a compound of claim 1 or a pharmaceutically
acceptable salt thereof and at least one pharmaceutically
acceptable excipient, carrier or diluent.
4. A product comprising a compound of claim 1 or a pharmaceutically
acceptable salt thereof and one or more chemotherapeutic agents, as
a combined preparation.
Description
The present invention provides novel substituted
4-carboxamido-isoindolinone derivatives which proved to be potent
and selective poly (ADP-ribose) polymerase-1 (PARP-1) inhibitors
with respect to poly (ADP-ribose) polymerase-2 (PARP-2) and are
thus useful in the therapy of cancer, cardiovascular diseases,
nervous system injury and inflammation. The present invention also
provides methods for preparing these compounds, pharmaceutical
compositions comprising these compounds, and methods of treating
diseases utilizing pharmaceutical compositions comprising these
compounds.
Poly (ADP-ribose) polymerases belong to a family of 18 members that
catalyze the addition of ADP-ribose units to DNA or different
acceptor proteins, which affect cellular processes as diverse as
replication, transcription, differentiation, gene regulation,
protein degradation and spindle maintenance. PARP-1 and PARP-2 are
the only enzymes among the PARPs that are activated by DNA damage
and are involved in DNA repair.
PARP-1 is a nuclear protein consisting of three domains: the
N-terminal DNA-binding domain containing two zinc fingers, the auto
modification domain, and the C-terminal catalytic domain. PARP-1
binds through the zinc-finger domain to DNA single strand breaks
(SSB), cleaves NAD.sup.+, and attaches multiple ADP-ribose units to
target proteins such as histones and various DNA repair enzymes.
This results in a highly negatively charged target, which in turn
leads to the unwinding and repair of the damaged DNA through the
base excision repair pathway. In knock out mouse models, deletion
of PARP-1 impairs DNA repair but it is not embryonic lethal. Double
knock out PARP-1 and PARP-2 mice instead die during early
embryogenesis, suggesting that the two enzymes display not
completely overlapping functions. Enhanced PARP-1 expression and/or
activity have been shown in different tumor cell lines, including
malignant lymphomas, hepatocellular carcinoma, cervical carcinoma,
colorectal carcinoma, leukemia. This may allow tumor cells to
withstand genotoxic stress and increase their resistance to
DNA-damaging agents. As a consequence, inhibition of PARP-1 through
small molecules has been shown to sensitize tumor cells to
cytotoxic therapy (e.g. temozolomide, platinums, topoisomerase
inhibitors and radiation). A significant window seems to exist
between the ability of a PARP inhibitor to potentiate therapeutic
benefits and undesirable side effects. Whereas the therapeutic use
of PARP inhibitors in combination with DNA damaging agents is not
novel, the use of these agents as monotherapy, in particular tumor
genetic backgrounds deficient in the homologous recombination DNA
repair, represents a new approach. Individuals with heterozygous
germ line mutations in either the BRCA-1 or BRCA-2 homologous
recombination repair genes exhibit high life time risks of
developing breast and other cancers. Tumors arising in mutation
carriers have generally lost the wild type allele and do not
express functional BRCA-1 and BRCA-2 proteins.
Therefore, loss of these two proteins leads to a tumor-specific
dysfunction in the repair of double strand breaks by homologous
recombination. It is known that when PARP-1 is inhibited, base
excision repair is reduced and single strand breaks that are
generated during the normal cell cycle persist. It has also been
established that replication forks that encounter an unrepaired
break can form double strand breaks which are normally repaired by
homologous recombination. Tumor cells that are deficient in
homologous recombination repair such as BRCA-1 and BRCA-2 mutants
are therefore highly sensitive to PARP inhibition compared with
wild-type cells. This is in line with the concept of synthetic
lethality, in which the two pathway defects alone are innocuous but
combined become lethal: PARP inhibitors may be more effective in
patients with tumors with specific DNA repair defects without
affecting normal heterozygous tissues. Putative patient population
includes, besides BRCA mutants that represent the majority of
hereditary breast and ovarian cancer, also a substantial fraction
of sporadic cancers with defects in homologous recombination
repair, a phenomenon termed "BRCAness". For example, methylation of
the promoters of the BRCA-1 or FANCF genes and amplification of the
EMSY gene, which encodes a BRCA-2 interacting protein. By extending
the rational of synthetic lethality of PARP and BRCA-1 and BRCA-2,
it is likely that deficiencies in any gene that is not redundant in
double strand break repair should be sensitive to PARP inhibition.
For example, ATM deficiency, found in patients with T-cell
prolymphocytic leukemia and B-cell chronic lymphocytic leukemia and
breast cancer and CHK2 germ line mutations identified in sarcoma,
breast cancer, ovarian cancer and brain tumors, have also been
shown to be synthetically lethal in combination with PARP
deficiency as well as deficiencies in other known HR pathway
proteins (including RAD51, DSS1, RAD54, RPA1, NBS1, ATR, CHK1,
CHK2, FANCD2, FANCA, FANCC and pTEN). Mutations in FANCC and FANCG
have been shown in pancreatic cancer. Methylation of FANCF promoter
has been found in ovarian, breast, cervical, lung carcinomas. The
first clinical evidence that BRCA-mutated cancer may be sensitive
to PARP inhibitor monotherapy comes from the phase I trial of the
oral, small molecule PARP inhibitor Olaparib. In an enriched phase
I population for BRCA mutation carriers, an objective response rate
of 47% were observed in 19 patients with BRCA mutations and breast,
ovarian and prostate cancer. Other PARP inhibitors, such as
Rucaparib and Veliparib are currently known to be in phase II
clinical trials in combination as well as single agent. Early
indications are that these therapies show low toxicity as single
agent. Anyway compounds with high selectivity on PARP-1 are
expected to show even less toxicity in view of a chronic treatment
schedule or in combination.
PARP-1 has also been implicated in angiogenesis. In particular,
PARP-1 inhibition seems to result in decreased accumulation of the
transcription hypoxia-inducible factor 1.alpha., an important
regulator of tumor cell adaptation to hypoxia.
Pro-inflammatory stimuli trigger the release of pro-inflammatory
mediators that induce the production of peroxynitrate and hydroxyl
radicals, which in turn yield to DNA single strand breakage with
consequent activation of PARP-1. Over activation of PARP-1 results
in depletion of NAD+ and energy stores, culminating in cell
dysfunction and necrosis. This cellular suicide mechanism has been
implicated in the pathomechanism of stroke, myocardial ischemia,
diabetes, diabetes-associated cardiovascular dysfunction, shock,
traumatic central nervous system injury, arthritis, colitis,
allergic encephalomyelitis and various other forms of inflammation.
Of special interest is the enhancement by PARP-1 of nuclear factor
kB-mediated transcription, which plays a central role in the
expression of inflammatory cytokines, chemokines and inflammatory
mediators.
WO 2007/047646 in the name of Janssen Pharmaceutica describes
substituted dihydro-isoindolones useful for treating kinase
disorders; Wender et al. claim in U.S. Pat. No. 7,232,842
isoindolone analogs as kinase inhibitors. The Patent Application US
2008/0108659 of Gandhi et al. describes
3-oxo-2,3-dihydro-1H-isoindoles as poly (ADP-ribose) polymerase
inhibitors, also reported in: Bioorg. Med. Chem. Lett., 2010, 20,
1023-1026. WO 2011/006794 and WO 2011/006803, both in the name of
Nerviano Medical Sciences, describe
3-oxo-2,3-dihydro-1H-isoindole-4-carboxamides as selective PARP-1
inhibitors.
The present invention provides novel substituted
4-carboxamido-isoindolinone derivatives which proved to be potent
and selective PARP-1 inhibitors with respect to PARP-2 and are thus
useful in the therapy of cancer, cardiovascular diseases, nervous
system injury and inflammation.
The present invention also provides method for preparing these
compounds, pharmaceutical compositions comprising these compounds,
and methods of treating diseases utilizing pharmaceutical
compositions comprising these compounds.
Accordingly, a first object of the present invention is to provide
a compound of formula (I):
##STR00001##
wherein:
R is hydrogen or fluorine; and
n, m, R1 and R2 have the following meanings:
a) n is 0 and m is 0, 1, 2 or 3;
R1 is 3- to 6-membered cycloalkyl or 4- to 6-membered heterocyclyl;
and
R2 is a 3-, 5- or 6-membered cycloalkyl, 4- to 6-membered
heterocyclyl, aryl or heteroaryl;
or
b) n is 1 and m is 0;
R1 is 3- to 6-membered cycloalkyl or aryl, each of which optionally
further substituted with one or more linear or branched
(C.sub.1-C.sub.6)-alkyl; and
R2 is null, 3- to 6-membered cycloalkyl, 4- to 6-membered
heterocyclyl, aryl or heteroaryl, each of which optionally further
substituted with one or more linear or branched
(C.sub.1-C.sub.6)-alkyl;
or
c) n is 2 or 3, and m is 0;
R1 is a 3- to 6-membered cycloalkyl, 4- to 6-membered heterocyclyl,
aryl or heteroaryl, each of which optionally further substituted
with one or more linear or branched (C.sub.1-C.sub.6)-alkyl;
and
R2 is null, 3- to 6-membered cycloalkyl, 4- to 6-membered
heterocyclyl, aryl or heteroaryl, each of which optionally further
substituted with one or more linear or branched
(C.sub.1-C.sub.6)-alkyl;
or
d) n and m are each independently 1, 2 or 3;
R1 and R2 are each independently 3- to 6-membered cycloalkyl, 4- to
6-membered heterocyclyl, aryl or heteroaryl;
or a pharmaceutically acceptable salt thereof.
The compounds of formula (I) as defined above are potent and
selective PARP-1 inhibitors with respect to PARP-2 and are thus
useful in cancer, cardiovascular diseases, nervous system injury
and inflammation therapy.
The present invention also provides methods of synthesizing
substituted 4-carboxamido-isoindolinone derivatives of formula (I)
as defined above, through a process consisting of standard
synthetic transformations.
The present invention also provides a method for treating diseases
mediated by PARP-1 protein which comprises administering to a
mammal in need thereof, preferably a human, an effective amount of
a compound of formula (I), as defined above.
A preferred method of the present invention is to treat a disease
mediated by PARP-1 protein selected from the group consisting of
cancer, cardiovascular diseases, nervous system injury and
inflammation.
Another preferred method of the present invention is to treat
specific types of cancer including, but not limited to: carcinomas,
such as bladder, breast, colon, kidney, liver, lung, including
small cell lung cancer, esophagus, gall-bladder, ovary, pancreas,
stomach, cervix, thyroid, prostate, and skin, including squamous
cell carcinoma; hematopoietic tumors of lymphoid lineage, including
leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia,
B-cell lymphoma, T-cell-lymphoma, Hodgkin's lymphoma, non-Hodgkin's
lymphoma, hairy cell lymphoma and Burkitt's lymphoma; hematopoietic
tumors of myeloid lineage, including acute and chronic myelogenous
leukemias, myelodysplastic syndrome and promyelocytic leukemia;
tumors of mesenchymal origin, including fibrosarcoma, Ewing's
sarcoma and rhabdomyosarcoma; tumors of the central and peripheral
nervous system, including astrocytoma, neuroblastoma, glioma and
schwannomas; and other tumors, including melanoma, seminoma,
teratocarcinoma, osteosarcoma, xeroderma pigmentosum,
keratoxanthoma, thyroid follicular cancer and Kaposi's sarcoma.
In addition, the method of the present invention also provides
tumor angiogenesis and metastasis inhibition.
Another preferred method of the present invention is to treat
specific types of cardiovascular diseases including, but not
limited to, myocardial reperfusion injury, cardiomyopathy, diabetic
cardiovascular dysfunction.
Another preferred method of the present invention is to treat
specific types of nervous system injury including but not limited
to: stroke, brain injury and neurodegenerative disorders.
Another preferred method of the present invention is to treat
specific types of inflammation diseases including, but not limited
to, colitis, arthritis and uveitis.
The present invention also provides an in vitro method for
selectively inhibiting PARP-1 protein activity which comprises
contacting the said protein with an effective amount of a compound
of formula (I), as defined above.
The present invention further provides a method for treating
diseases comprising a compound of formula (I), as defined above, in
combination with radiation therapy or chemotherapy regimen for
simultaneous, separate or sequential use in anticancer therapy.
The present invention also provides a pharmaceutical composition
comprising a therapeutically effective amount of a compound of
formula (I) or a pharmaceutically acceptable salt thereof, as
defined above, and at least one pharmaceutically acceptable
excipient, carrier or diluent.
In addition to a compound of formula (I), the pharmaceutical
composition of the present invention may further comprise one or
more chemotherapeutic--e.g. cytostatic or cytotoxic--agents,
antibiotic-type agents, alkylating agents, antimetabolite agents,
hormonal agents, immunological agents, interferon-type agents,
cyclooxygenase inhibitors (e.g. COX-2 inhibitors), matrix
metalloproteinases inhibitors, telomerase inhibitors, tyrosine
kinase inhibitors, anti-growth factor receptor agents, anti-HER
agents, anti-EGFR agents, anti-angiogenesis agents (e.g.
angiogenesis inhibitors), farnesyl transferase inhibitors, ras-raf
signal transduction pathway inhibitors, cell cycle inhibitors,
other cdks inhibitors, tubulin binding agents, topoisomerase I
inhibitors, topoisomerase II inhibitors, and the like. Preferably,
the chemotherapeutic agent is an alkylating agent. Even more
preferably, the alkylating agent is temozolomide.
Additionally, the invention provides a product comprising a
compound of formula (I) or a pharmaceutically acceptable salt
thereof, as defined above, and one or more chemotherapeutic agents,
as a combined preparation for simultaneous, separate or sequential
use in anticancer therapy. Preferably, the chemotherapeutic agent
is an alkylating agent. Even more preferably, the alkylating agent
is temozolomide.
Moreover, the invention provides a compound of formula (I) or a
pharmaceutically acceptable salt thereof, as defined above, for use
as a medicament, preferably as a medicament with anticancer
activity.
In yet another aspect, the invention provides a compound of formula
(I) or a pharmaceutically acceptable salt thereof, as defined
above, for use in a method of treating cancer.
Finally, the invention provides the use of a compound of formula
(I) or a pharmaceutically acceptable salt thereof, as defined
above, in the manufacture of a medicament with anticancer
activity.
The compounds of formula (I) may have one or more asymmetric
centers, and may therefore exist as individual optical isomers or
racemic mixtures or diastereoisomers. Accordingly, all the possible
isomers, and their mixtures of the compounds of formula (I) are
within the scope of the present invention. As stated above, salts
of the compounds of formula (I) are also within the scope of the
present invention.
Unless otherwise specified, when referring to the compounds of
formula (I) per se as well as to any pharmaceutical composition
thereof or to any therapeutic treatment comprising them, the
present invention includes all of the isomers, tautomers, hydrates,
solvates, N-oxides and pharmaceutically acceptable salts of the
compounds of this invention.
If a chiral center or another form of an isomeric center is present
in a compound of the present invention, all forms of such isomer or
isomers, including enantiomers and diastereoisomers, are intended
to be covered herein. Compounds containing a chiral center may be
used as a racemic mixture, an enantiomerically enriched mixture, or
the racemic mixture may be separated using well-known techniques
and an individual enantiomer may be used alone. In cases in which
compounds have unsaturated carbon-carbon double bonds, both the cis
(Z) and trans (E) isomers are within the scope of this
invention.
In cases wherein compounds may exist in tautomeric forms, such as
keto-enol tautomers, each tautomeric form is contemplated as being
included within this invention whether existing in equilibrium or
predominantly in one form.
With the term "halogen" we intend a fluorine, chlorine, bromine or
iodine atom.
With the term "linear or branched (C.sub.1-C.sub.6)-alkyl", we
intend any of the groups such as, for instance, methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl,
n-pentyl, n-hexyl, and the like.
With the term "3- to 6-membered cycloalkyl" we intend, unless
otherwise provided, a 3- to 6-membered all-carbon monocyclic ring,
which may contain one or more double bonds but does not have a
completely conjugated 7-electron system. Examples of cycloalkyl
groups, without limitation, are cyclopropyl, cyclobutyl,
cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl and
cyclohexadienyl.
With the term "4- to 6-membered heterocyclyl" we intend a 4- to
6-membered, saturated or partially unsaturated carbocyclic ring
where one or more carbon atoms are replaced by heteroatoms such as
nitrogen, oxygen and sulfur; the heterocyclyl ring can be
optionally further fused or linked to aromatic and non-aromatic
carbocyclic and heterocyclic rings. Non limiting examples of
heterocyclyl groups are, for instance, pyranyl, pyrrolidinyl,
pyrrolinyl, imidazolinyl, imidazolidinyl, pyrazolidinyl,
pyrazolinyl, thiazolinyl, thiazolidinyl, dihydrofuranyl,
tetrahydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl,
morpholinyl and the like.
The term "aryl" refers to a mono-, bi- or poly-carbocyclic
hydrocarbon with from 1 to 4 ring systems, optionally further fused
or linked to each other by single bonds, wherein at least one of
the carbocyclic rings is "aromatic", wherein the term "aromatic"
refers to completely conjugated .pi.-electron bond system. Non
limiting examples of such aryl groups are phenyl, .alpha.- or
.beta.-naphthyl or biphenyl groups.
The term "heteroaryl" refers to aromatic heterocyclic rings,
typically 5- to 8-membered heterocycles with from 1 to 3
heteroatoms selected among N, O or S; the heteroaryl ring can be
optionally further fused or linked to aromatic and non-aromatic
carbocyclic and heterocyclic rings. Not limiting examples of such
heteroaryl groups are, for instance, pyridyl, pyrazinyl,
pyrimidinyl, pyridazinyl, indolyl, imidazolyl, thiazolyl,
isothiazolyl, pyrrolyl, phenyl-pyrrolyl, furyl, phenyl-furyl,
oxazolyl, isoxazolyl, pyrazolyl, thienyl, benzothienyl,
isoindolinyl, benzoimidazolyl, indazolyl, quinolinyl,
isoquinolinyl, 1,2,3-triazolyl, 1-phenyl-1,2,3-triazolyl,
2,3-dihydroindolyl, 2,3-dihydrobenzofuranyl,
2,3-dihydrobenzothiophenyl; benzopyranyl, 2,3-dihydrobenzoxazinyl,
2,3-dihydroquinoxalinyl and the like.
According to the present invention and unless otherwise provided,
when any of the above mentioned groups is optionally substituted,
it may be substituted in any of its free position by one or more
linear or branched (C.sub.1-C.sub.6) alkyl groups.
The term "pharmaceutically acceptable salt" of compounds of formula
(I) refers to those salts that retain the biological effectiveness
and properties of the parent compound, therefore pharmaceutically
acceptable salts of the compounds of formula (I) include the acid
addition salts with inorganic or organic acids, e.g., nitric,
hydrochloric, hydrobromic, sulfuric, perchloric, phosphoric,
acetic, trifluoroacetic, propionic, glycolic, (D) or (L) lactic,
oxalic, ascorbic, fumaric, malonic, malic, maleic, tartaric,
citric, benzoic, cinnamic, mandelic, methanesulphonic,
ethanesulfonic, p-toluenesulfonic, isethionic, succinic and
salicylic acid.
Pharmaceutically acceptable salts of the compounds of formula (I)
also include the salts with inorganic or organic bases, e.g.,
alkali or alkaline-earth metals, especially sodium, potassium,
calcium, ammonium or magnesium hydroxides, carbonates or
bicarbonates, and acyclic or cyclic amines, preferably methylamine,
ethylamine, diethylamine, triethylamine, piperidine and the
like.
In a first preferred embodiment, the present invention provides
compounds of formula (I) as defined above characterized in that
R is hydrogen or fluorine; and
n, m, R1 and R2 have the following meanings:
a) n is 0 and m is 0 or 1;
R1 is a 6-membered heterocyclyl; and
R2 is a 3- or 6-membered cycloalkyl, 6-membered heterocyclyl, aryl
or heteroaryl;
or
b) n is 1 and m is 0;
R1 is aryl, optionally further substituted with one or more linear
or branched (C.sub.1-C.sub.6)-alkyl; and
R2 is null;
or
c) n is 2 or 3, and m is 0;
R1 is a 6-membered heterocyclyl, aryl or heteroaryl, each of which
optionally further substituted with one or more linear or branched
(C.sub.1-C.sub.6)-alkyl; and
R2 is null, a 6-membered heterocyclyl or aryl;
or
d) n is 2 or 3, and m is 1;
R1 is a 6-membered heterocyclyl; and
R2 is aryl;
or a pharmaceutically acceptable salt thereof.
In a more preferred embodiment, the present invention provides
compounds of formula (I) as defined above characterized in that
R is hydrogen or fluorine; and
n, m, R1 and R2 have the following meanings:
a) n is 0 and m is 0 or 1;
R1 is a 6-membered heterocyclyl; and
R2 is a 3- or 6-membered cycloalkyl, 6-membered heterocyclyl, aryl
or heteroaryl;
or
c) n is 2 or 3, and m is 0;
R1 is a 6-membered heterocyclyl, aryl or heteroaryl, each of which
optionally further substituted with one or more linear or branched
(C.sub.1-C.sub.6)-alkyl; and
R2 is null, a 6-membered heterocyclyl or aryl;
or a pharmaceutically acceptable salt thereof.
Even more preferably, the present invention provides compounds of
formula (I) as defined above characterized in that:
R is hydrogen or fluorine; and
n, m, R1 and R2 have the following meanings:
a) n is 0 and m is 0 or 1;
R1 is a 6-membered heterocyclyl; and
R2 is a 3- or 6-membered cycloalkyl, 6-membered heterocyclyl, aryl
or heteroaryl;
or a pharmaceutically acceptable salt thereof.
Most preferably, the present invention provides compounds of
formula (I) as defined above characterized in that:
R is hydrogen or fluorine; and
n, m, R1 and R2 have the following meanings:
a) n is 0 and m is 0 or 1;
when m is 0, R1 is a piperidine ring and R2 is a cyclohexyl
ring;
when m is 1, R1 is a piperidine ring and R2 is a pyridine ring;
or a pharmaceutically acceptable salt thereof.
Specific preferred compounds (cpd) of the present invention are
listed below: 1.
2-benzyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic acid amide; 2.
3-oxo-2-phenethyl-2,3-dihydro-1H-isoindole-4-carboxylic acid amide;
3. 2-[2-(3,4-dihydro-1H-isoquinolin-2-yl)-ethyl]-3-oxo-2,3-dihydro-
-1H-isoindole-4-carboxylic acid amide; 4.
3-oxo-2-(2-piperidin-1-yl-ethyl)-2,3-dihydro-1H-isoindole-4-carboxylic
acid amide; 5.
2-(2-morpholin-4-yl-ethyl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic
acid amide; 6.
2-(3-morpholin-4-yl-propyl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic
acid amide; 7.
2-[2-(3,4-dihydro-2H-quinolin-1-yl)-ethyl]-3-oxo-2,3-dihydro-1H-isoindole-
-4-carboxylic acid amide; 8.
3-oxo-2-(1-pyridin-4-ylmethyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4--
carboxylic acid amide; 9.
3-oxo-2-(1-thiophen-2-ylmethyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-
-carboxylic acid amide; 10.
3-oxo-2-(1-pyridin-3-ylmethyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4--
carboxylic acid amide; 11.
2-(1-cyclohexyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxyl-
ic acid amide; 12.
2-(1-furan-2-ylmethyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-ca-
rboxylic acid amide; 13.
3-oxo-2-(1-thiophen-3-ylmethyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-
-carboxylic acid amide; 14.
2-(1-furan-3-ylmethyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-ca-
rboxylic acid amide; 15.
3-oxo-2-(1-pyridin-2-ylmethyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4--
carboxylic acid amide; 16.
3-oxo-2-[1-(1H-pyrrol-2-ylmethyl)-piperidin-4-yl]-2,3-dihydro-1H-isoindol-
e-4-carboxylic acid amide; 17.
3-oxo-2-(3-phenyl-propyl)-2,3-dihydro-1H-isoindole-4-carboxylic
acid amide; 18.
3-oxo-2-(2-pyridin-2-yl-ethyl)-2,3-dihydro-1H-isoindole-4-carboxylic
acid amide; 19.
2-[3-(3,4-dihydro-1H-isoquinolin-2-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoin-
dole-4-carboxylic acid amide; 20.
2-[3-(3,4-dihydro-2H-quinolin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindol-
e-4-carboxylic acid amide; 21.
2-[3-(4-methyl-piperazin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole-4-c-
arboxylic acid amide; 22.
3-oxo-2-[3-(4-phenyl-piperazin-1-yl)-propyl]-2,3-dihydro-1H-isoindole-4-c-
arboxylic acid amide; 23.
6-fluoro-2-(3-morpholin-4-yl-propyl)-3-oxo-2,3-dihydro-1H-isoindole-4-car-
boxylic acid amide; 24.
2-(1-cyclopropylmethyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-c-
arboxylic acid amide; 25.
3-oxo-2-(3-piperidin-1-yl-propyl)-2,3-dihydro-1H-isoindole-4-carboxylic
acid amide; 26.
2-(3-[1,4]bipiperidinyl-1-yl-propyl)-3-oxo-2,3-dihydro-1H-isoindole-4-car-
boxylic acid amide; 27.
2-[3-(2,6-dimethyl-piperidin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole-
-4-carboxylic acid amide; 28.
3-oxo-2-[1-(tetrahydro-pyran-4-yl)-piperidin-4-yl]-2,3-dihydro-1H-isoindo-
le-4-carboxylic acid amide; 29.
2-(1-cyclohexyl-piperidin-4-yl)-6-fluoro-3-oxo-2,3-dihydro-1H-isoindole-4-
-carboxylic acid amide; 30.
2-(1-benzyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic
acid amide; 31.
2-[2-(1-benzyl-piperidin-4-yl)-ethyl]-3-oxo-2,3-dihydro-1H-isoindole-4-ca-
rboxylic acid amide; 32.
2-[3-(4-benzyl-piperidin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole-4-c-
arboxylic acid amide; 33.
2-[1-(4,4-dimethyl-cyclohexyl)-piperidin-4-yl]-3-oxo-2,3-dihydro-1H-isoin-
dole-4-carboxylic acid amide; 34.
2-[1-(4,4-dimethyl-cyclohexyl)-piperidin-4-yl]-6-fluoro-3-oxo-2,3-dihydro-
-1H-isoindole-4-carboxylic acid amide; 35.
6-fluoro-3-oxo-2-(1-spiro[2.5]oct-6-yl-piperidin-4-yl)-2,3-dihydro-1H-iso-
indole-4-carboxylic acid amide; 36.
3-oxo-2-(1-spiro[2.5]oct-6-yl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4--
carboxylic acid amide;
or a pharmaceutically acceptable salt thereof.
The present invention also provides processes for the preparation
of compounds of formula (I) as defined above. Accordingly, a
process of the present invention comprises one of the following
sequences of steps:
Sequence A (when R is Fluorine, Scheme 1):
either
Step a) halogenating 4-fluoro-2-methyl-phenylamine (XI):
##STR00002##
Step b) cyano-de-aminating the resultant compound of formula
(X):
##STR00003##
wherein Hal is halogen such as Cl, Br, and I;
Step c) hydrolyzing the resultant compound of formula (IX):
##STR00004##
wherein Hal is as defined above and
Step d) hydrolyzing the resultant compound of formula (VIII):
##STR00005##
wherein Hal is as defined above;
or
Step e) halogenating 4-fluoro-2-methyl-benzoic acid (XII):
##STR00006##
Then:
Step f) esterifying the compound of formula (VII) obtained in step
d) or e)
##STR00007##
wherein Hal is as defined above;
Step g) cyano-de-halogenating the resultant compound of formula
(VI):
##STR00008##
wherein T is a (C.sub.1-C.sub.6)-alkyl or an
aryl-(C.sub.1-C.sub.6)-alkyl and Hal is as defined above;
Step h) cyclizing the resultant compound of formula (V):
##STR00009##
wherein T is as defined above, by reaction with a suitable amine of
formula (XIII) X--R1-[CH.sub.2].sub.n--NH.sub.2 (XIII)
wherein R1 and n are as defined above, and X is either
R2-[CH.sub.2].sub.m--, wherein R2 and m are as defined above, or a
suitable nitrogen protective group, when R1 is a nitrogen
containing heterocyclyl;
Step c') hydrolyzing the resultant compound of formula (IV):
##STR00010##
wherein R1, n and X are as defined above, so as to obtain
a compound of formula (I), as defined above, when X is
R2-[CH.sub.2].sub.m--, wherein R2 and m are as defined above;
or
a compound of formula (III), when R1 is a nitrogen containing
heterocyclyl and X is a suitable nitrogen protective group,
##STR00011##
wherein n is as defined above, R1 is a nitrogen containing
heterocyclyl and X is a suitable nitrogen protective group;
Step i) deprotecting the compound of formula (III), as defined
above, so as to obtain either
a compound of formula (I), as defined above, or
a compound of formula (II):
##STR00012##
wherein R1 and n are as defined above;
Step l) alkylating the resultant compound of formula (II), as
defined above, with a suitable alkylating agent of formula (XIV)
R2-[CH.sub.2].sub.m-1--Y (XIV)
wherein Y is either a formyl group or, when m=1, an oxygen atom
linked to R2 through a double bond (.dbd.O), so as to obtain a
compound of formula (I).
Sequence B (when R is Hydrogen, Scheme 2):
Step m) performing a reductive amination on furan-2-carbaldehyde
(XV):
##STR00013##
with a suitable amine of formula (XIII)
X--R1-[CH.sub.2].sub.m--NH.sub.2 (XIII)
wherein R1 and n are as defined above, and X is either
R2-[CH.sub.2].sub.m--, wherein R2 and m are as defined above, or a
suitable nitrogen protective group, when R1 is a nitrogen
containing heterocyclyl;
Step n) performing a Diels-Alder reaction on the resultant compound
of formula (XVI):
##STR00014##
wherein R1, n and X are as defined above;
Step o) aromatizing the resultant compound of formula (XVII):
##STR00015##
wherein R1, n and X are as defined above;
Step p) amidating the resultant compound of formula (XVIII):
##STR00016##
wherein R1, n and X are as defined above, so as to obtain
a compound of formula (I), as defined above, when X is
R2-[CH.sub.2].sub.m--, wherein R2 and m are as defined above;
or
a compound of formula (XX), when R1 is a nitrogen containing
heterocyclyl and X is a suitable nitrogen protective group,
##STR00017##
wherein n is as defined above, R1 is a nitrogen containing
heterocyclyl and X is a suitable nitrogen protective group;
Step i') deprotecting a compound of formula (XX) as defined
above;
Step l') alkylating the resultant compound of formula (XXI):
##STR00018##
wherein R1 and n are as defined above, with a suitable alkylating
agent of formula (XIV) R2-[CH.sub.2].sub.m-1--Y (XIV)
wherein Y is either a formyl group or, when m=1, an oxygen atom
linked to R2 through a double bond (.dbd.O), so as to obtain a
compound of formula (I), as defined above.
In case, during Step o, the compound resulting from aromatization
of a compound of formula (XVII) is a compound of formula (XIX),
i.e. when X is a labile nitrogen protective group, the following
Step q is performed:
Step q) installing a suitable nitrogen protective group on the
resultant compound of formula (XIX):
##STR00019##
wherein R1 and n are as defined above, so as to obtain a compound
of formula (XVIII), wherein R1 and n are as defined above and X is
a suitable nitrogen protective group, which is then subjected to
the sequence of reactions p), i') and l') above described so as to
obtain a compound of formula (I) as above defined.
If necessary or wanted, the processes above described comprises
converting a compound of formula (I) into a different compound of
formula (I) by known chemical reactions; and/or, if desired,
converting a compound of formula (I) into a pharmaceutically
acceptable salt thereof or converting a salt into a free compound
of formula (I).
Such known chemical reactions for possible conversions of compounds
into different compounds comprise for instance a reductive
amination (Cv1).
All the above processes are analogy processes which can be carried
out according to well known methods and under suitable conditions
known in the art.
The synthesis of a compound of formula (I), according to the
synthetic processes described above, can be carried out in a
stepwise manner, whereby each intermediate is isolated and purified
by standard purification techniques, like, for example, column
chromatography, before carrying out the subsequent reaction.
Alternatively, two or more steps of the synthetic sequence can be
carried out in a so-called "one-pot" procedure, as known in the
art, whereby only the compound resulting from the two or more steps
is isolated and purified.
Schemes 1-2 below show the preparation of a compound of formula (I)
as defined above.
##STR00020##
##STR00021##
According to step a), a compound of formula (X) can be obtained by
halogenating 4-fluoro-2-methyl-phenylamine (XI) in a variety of
ways and experimental conditions known in the art. Preferably this
reaction is conducted in the presence of N-bromosuccinimide,
N-iodosuccinimmide, N-chlorosuccinimide, bromine, iodine,
hydrobromic acid/hydrogen peroxide, in a suitable solvent, such as
acetonitrile, N,N-dimethylformamide, dioxane, dimethylsulfoxide,
acetic acid or water, at a temperature ranging from about room
temperature to reflux and for a period of time varying from about 1
h to about 96 h.
According to step b), a compound of formula (IX) can be obtained by
a two-steps reaction sequence from a compound of formula (X) in a
variety of ways and experimental conditions known in the art. First
step is preferably conducted in the presence of sodium
nitrite/hydrochloric acid or tert-butylnitrite in a suitable
solvent, such as tetrahydrofuran, dimethoxyethane,
dimethylsulfoxide, acetic acid or water, at a temperature ranging
from about -20.degree. C. to room temperature and for a period of
time varying from 10 min to about 24 h. Second step is preferably
carried out in the presence of sodium, copper or potassium cyanide,
often in the presence of an additive such as copper or potassium
chloride, in a suitable solvent, such as tetrahydrofuran,
dimethoxyethane, dimethylsulfoxide, acetic acid, toluene or water,
at a temperature ranging from about -20.degree. C. to reflux and
for a period of time ranging from about 10 min to about 96 h.
According to step c), the hydrolysis of a compound of formula (IX)
to a give a compound of formula (VIII) can be carried out in a
variety of ways, according to conventional methods for transforming
a cyano group to amide. Preferably this reaction is carried out in
a suitable solvent such as, for instance, methanol, ethanol,
butanol, 1,4-dioxane, toluene, water, or a mixture thereof, in the
presence of a suitable acid or base, such as, for instance,
sulfuric acid, methanesulfonic acid, hydrochloric acid,
trifluoroacetic acid, sodium hydroxide, sodium carbonate, or a
suitable reagent such as hydrogen peroxide, sodium perborate or
indium(III) salts in the presence of acetaldoxime. Typically, the
reaction is carried out at a temperature ranging from room
temperature to reflux and for a time varying from about 1 h to
about 96 h.
According to step d), a compound of formula (VIII) can be
transformed into a compound of formula (VII) according to
conventional methods. Preferably the reaction is carried out in the
presence of water by treatment with a base such as potassium or
sodium carbonate, potassium or sodium hydroxide, in a suitable
solvent such as, for instance, methanol or ethanol, at a
temperature ranging from room temperature to reflux, for a time
ranging from about 30 min to about 96 h. Alternatively this
reaction can be conducted in the presence of sodium nitrite/acetic
acid, sulfuric acid, phosphoric acid, at a temperature ranging from
room temperature to reflux and for a time varying from about 1 h to
about 96 h.
According to step e), the halogenation of 4-fluoro-2-methyl-benzoic
acid (XII) into a compound of formula (VII) can be carried out in a
variety of ways, according to conventional methods for halogenation
reactions. Preferably, this reaction is carried out with
tetrabutylammonium bromide and/or iodine in the presence of
phenyliodine(III) bis(trifluoracetate) or phenyliodo(III) diacetate
as halogen source in a suitable solvent such as, for instance,
N,N-dimethylformamide or dichloroethane, at a temperature ranging
from room temperature to reflux and for a time varying from about 1
h to about 48 h. The catalyst is usually a metal, most often a
palladium derivative such as, for instance, palladium(II) chloride
or palladium(II) acetate.
According to step f), a compound of formula (VII) can be
transformed into a compound of formula (VI) according to
conventional methods. Preferably the reaction is carried out in the
presence of hydrochloric acid, sulfuric acid or acetic acid by
using as a solvent methanol, ethanol, water, or a mixture thereof,
at a temperature ranging from room temperature to reflux and for a
time varying from about 1 h to about 96 h. Alternatively, this
reaction can be conducted with alkyl iodide, bromide or
toluensulfonate in the presence of a suitable base, such as sodium
or potassium carbonate, and sodium, lithium or potassium hydroxide,
at a temperature ranging from room temperature to reflux and for a
time varying from about 1 h to about 96 h.
According to step g), the transformation of a compound of formula
(VI) into a compound of formula (V) can be carried out in a variety
of ways, according to conventional methods for cyanation reactions.
Preferably, this reaction is carried out in the presence of
copper(I) cyanide or potassium hexacyanoferrate(II) as cyano source
in a suitable solvent such as, for instance, methanol, ethanol,
tetrahydrofuran, 1,4-dioxane, toluene, xylene,
N-methyl-2-pyrrolidone, N,N-dimethylformamide,
N,N-dimethylacetamide or a mixture thereof, at a temperature
ranging from room temperature to reflux and for a time varying from
about 1 h to about 96 h. If a catalyst is required, it is usually a
metal, most often a palladium derivative such as, for instance,
tetrakis(triphenylphosphine)palladium(0), palladium(II) chloride or
palladium(II) acetate in the presence of a suitable base such as,
for instance, sodium, potassium or cesium carbonate or cesium
fluoride.
According to step h), a compound of formula (IV) can be obtained by
a two-steps reaction sequence from a compound of formula (V) in the
presence of a compound of formula (XIII) in a variety of ways and
experimental conditions known in the art. First step is preferably
conducted in the presence of N-bromosuccinimide with a radical
initiator such as benzoyl peroxide or azobisisobutyronitrile in a
suitable solvent, such as carbon tetrachloride, chloroform,
dichloromethane or methyl pivalate, at a temperature ranging from
about room temperature to reflux and for a period of time varying
from 10 min to about 24 h. Second step can be conducted both under
basic or acidic conditions, such as in the presence of sodium or
potassium carbonate, 1,8-diazabicyclo[5.4.0]undec-7-ene,
triethylamine, diisopropylethylamine, pyridine or acetic acid,
hydrochloric acid, in a suitable solvent, such as tetrahydrofuran,
dimethoxyethane, 1,4-dioxane or toluene, at a temperature ranging
from room temperature to reflux and for a period of time varying
from 1 h to about 96 h.
According to step c'), the hydrolysis of a compound of formula (IV)
to give either a compound of formula (I) or a compound of formula
(III), can be carried out in a variety of ways and experimental
conditions. Preferably it is carried out in a way analogous to that
reported for step c).
According to step i), when, in a compound of formula (III), X is a
nitrogen protective group such as tert-butoxycarbonyl,
4-methoxybenzyl, 2,4-dimethoxybenzyl and triphenylmethyl protective
groups, either a compound of formula (I) or a compound of formula
(II) can be obtained by removing these protective groups under
acidic conditions, preferably in the presence of an inorganic or
organic acid such as hydrochloric, trifluoroacetic or
methanesulphonic acid, boron tribromide or aluminium trichloride,
in a suitable solvent, such as dichloromethane, dichloroethane,
dioxane or a lower alcohol, such as methanol or ethanol, at a
temperature ranging from room temperature to reflux. When, in a
compound of formula (III), X is a nitrogen protective group such as
benzyloxycarbonyl and the like, either a compound of formula (I) or
a compound of formula (II) can be obtained by removing these
protective groups under reducing conditions, such as, for instance,
in the presence of hydrogen and a hydrogenation catalyst in a
suitable solvent, such as ethanol, methanol, ethyl acetate, or a
mixture thereof. The catalyst is usually a metal, most often a
palladium derivative such as, for instance, palladium on carbon,
palladium hydroxide or palladium black. When, in a compound of
formula (III), X is a nitrogen protective group such as
methoxycarbonyl, ethoxycarbonyl, 9-fluorenylmethoxycarbonyl and the
like, either a compound of formula (I) or a compound of formula
(II) can be obtained by removing these protective groups under
basic conditions such as, for instance, sodium, potassium or cesium
carbonate, sodium, potassium or barium hydroxide, hydrazine,
piperidine, morpholine or the like, in a suitable solvent, such as
methanol, ethanol, water, N,N-dimethylformamide,
N,N-dimethylacetamide or the like, at a temperature ranging from
room temperature to reflux.
According to step l), the reductive alkylation of a compound of
formula (II), in the presence of a compound of formula (XIV), to
give a compound of formula (I), can be conducted in a variety of
ways, according to conventional methods for carrying out reductive
amination. Preferably, this reaction is carried out in a suitable
solvent such as, for instance, methanol, N,N-dimethylformamide,
dichloromethane, tetrahydrofuran, benzene, toluene, or a mixture
thereof, in the presence of a suitable reducing agent such as, for
instance, sodium borohydride, tetraalkylammonium borohydride,
sodium cyano borohydride, sodium triacetoxyborohydride,
tetramethylammonium triacetoxy borohydride and in the presence of
an acid or basic catalyst, such as, for instance, acetic acid,
trifluoroacetic acid, zinc chloride, zinc bromide, tin(IV)
chloride, titanium(IV) chloride, boron trifluoride or
triethylamine, diisopropylethylamine or pyridine, at a temperature
ranging from about 0.degree. C. to reflux and for a time varying
from about 1 h to about 96 h.
According to step m), a compound of formula (XVI) can be obtained
from furan-2-carbaldehyde (XV) through reductive amination in the
presence of a compound of formula (XIII). Preferably, this reaction
is carried out in a suitable solvent such as, for instance,
methanol, N,N-dimethylformamide, dichloromethane, tetrahydrofuran,
benzene, toluene, or a mixture thereof, in the presence of a
suitable reducing agent such as, for instance, sodium borohydride,
tetraalkylammonium borohydride, sodium cyano borohydride, sodium
triacetoxyborohydride or tetramethylammonium triacetoxy
borohydride, and in the presence of an acid or basic catalyst, such
as, for instance, acetic acid, trifluoroacetic acid, zinc chloride,
zinc bromide, tin(IV) chloride, titanium(IV) chloride, boron
trifluoride or triethylamine, diisopropylethylamine or pyridine, at
a temperature ranging from about 0.degree. C. to reflux and for a
time varying from about 1 h to about 96 h.
According to step n), the Diels-Alder reaction, performed on a
compound of formula (XVI) to give a compound of formula (XVII), can
be conducted in a variety of ways, according to conventional
methods for carrying out these reactions. Preferably, this reaction
is carried out in a suitable solvent such as, for instance,
tetrahydrofuran, benzene, toluene or o-xylene, in the presence of
maleic anhydride at a temperature ranging from about room
temperature to reflux and for a time varying from about 1 h to
about 96 h.
According to step o), the transformation of a compound of formula
(XVII) into either a compound of formula (XVIII) or a compound of
formula (XIX), can be carried out in a variety of ways, according
to conventional methods. Preferably, this reaction is carried out
in a suitable solvent such as, for instance, tetrahydrofuran,
toluene or water, in the presence of hydrochloric acid,
p-toluenesulfonic acid or phosphoric acid, at a temperature ranging
from about room temperature to reflux and for a time varying from
about 1 h to about 24 h.
According to step p), a compound of formula (XVIII) can be reacted
either to deliver a compound of formula (I) or a compound of
formula (XX) in a variety of ways and experimental conditions,
which are widely known in the art of condensation reactions.
Preferably a compound of formula (XVIII) is reacted with ammonia or
ammonia source such as ammonium salts, in the presence of an
activating agent such as carbonyldiimidazole,
benzotriazol-1-yloxy)tris(dimethylamino)phosphonium
hexafluorophosphate,
2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium
tetrafluoroborate),
(O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate, dicyclohexyl carbodiimide, diisopropyl
carbodiimide, 1-ethyl-3-(3'-dimethylamino) carbodiimide
hydrochloric acid salt, optionally in the presence of
hydroxybenzotriazole. Preferably, this reaction is carried out in a
suitable solvent such as, for instance, N,N-dimethylformamide,
N,N-dimethylacetamide, tetrahydrofuran, dichloromethane or
1,4-dioxane, and in the presence of a proton scavenger such as, for
example, pyridine, triethylamine or diisopropylethylamine, at a
temperature ranging from room temperature to reflux, for a time
ranging from about 30 min to about 96 h.
According to step i'), the deprotection of a compound of formula
(XX) to give a compound of formula (XXI), can be carried out in a
variety of ways and experimental conditions. Preferably it is
carried out in a way analogous to that reported for step i).
According to step l') the reductive alkylation of a compound of
formula (XXI), in the presence of a compound of formula (XIV), to
give a compound of formula (I), can be carried out in a variety of
ways and experimental conditions. Preferably it is carried out in a
way analogous to that reported for step l).
According to step q), the protection of compound of formula (XIX)
to give a compound of formula (XVIII), where X is a suitable
nitrogen protective group, may be carried out in a variety of ways
and experimental conditions. Preferably, when the protective group
is tert-butoxycarbonyl, the reaction may be carried out in the
presence of di-tert-butyl dicarbonate in a variety of solvents such
as methanol, ethanol, acetonitrile, tetrahydrofuran or
dichloromethane, in the presence of a base, such as pyridine,
N,N-dimethylaminopyridine, triethylamine, diisopropylethylamine,
sodium or potassium carbonate, at a temperature ranging from room
temperature to reflux and for a time varying from about 1 h to
about 96 h.
According to the conversion 1 (Cv1), the reductive alkylation of a
compound of formula (I) to give another compound of formula (I) may
be carried out in a variety of ways and experimental conditions.
Preferably, it is carried out in a way analogous to that reported
for step l).
Substituted isoindolinone derivatives can be prepared using
standard procedures in organic synthesis as reported, for instance,
in Smith, Michael--March's Advanced Organic Chemistry: reactions
mechanisms and structure--6.sup.th Edition, Michael B. Smith and
Jerry March, John Wiley & Sons Inc., New York (N.Y.), 2007. It
is known to the skilled person that transformation of a chemical
function into another may require that one or more reactive centers
in the compound containing this function have to be protected in
order to avoid undesired side reactions. Protection of such
reactive centers, and subsequent deprotection at the end of the
synthetic transformations, can be accomplished following standard
procedures described, for instance, in: Green, Theodora W. and
Wuts, Peter G. M.--Protective Groups in Organic Synthesis, Third
Edition, John Wiley & Sons Inc., New York (N.Y.), 1999.
In cases where a compound of formula (I) contains one or more
asymmetric centers, said compound can be separated into the single
isomers by procedures known to those skilled in the art. Such
procedures comprise standard chromatographic techniques, including
chromatography using a chiral stationary phase, or crystallization.
General methods for separation of compounds containing one or more
asymmetric centers are reported, for instance, in Jacques, Jean;
Collet, Andre; Wilen, Samuel H.--Enantiomers, Racemates, and
Resolutions, John Wiley & Sons Inc., New York (N.Y.), 1981.
A compound of formula (I) can also be transformed into a
pharmaceutically acceptable salt according to standard procedures
that are known to those skilled in the art. Alternatively, a
compound of formula (I) that is obtained as a salt can be
transformed into the free base or the free acid according to
standard procedures that are known to the skilled person.
The starting materials of the process of the present invention,
i.e. 4-fluoro-2-methyl-phenylamine (XI), 4-fluoro-2-methyl-benzoic
acid (XII), furan-2-carbaldehyde (XV) and compounds of formula
(XIII) and (XIV) are either commercially available or can be
prepared by using well-known methods.
Pharmacology
PARP-1 is a DNA damage-induced polymerase that catalyzes the
cleavage of NAD+ into nicotinamide and ADP-ribose and then uses the
latter to synthesize branched nucleic-acid like poly(ADP-ribose)
polymers. In vivo, the most abundantly poly (ADP-ribosylated)
protein is PARP-1 itself, followed by histones. PARP-1 is
responsible for 90% of this DNA damage-induced activity while the
remaining 10% is due to PARP-2.
Biochemical Assay
Affinity evaluation of the tested compounds and their selectivity
with respect to the different PARP isoforms of interest was
assessed in a displacement assay.
The identification of compounds capable of binding several PARP
proteins is carried out through a screening method including the
steps of
a) providing a reaction mixture containing:
the PARP protein isoform under investigation,
a compound of formula (IP):
##STR00022##
wherein R.sub.11 is hydrogen or a methyl group, B is
(CH.sub.2).sub.n--NH group wherein n is 2 to 6; m is 0 or 1 and
X.sup.--- is a counterion, and
serial dilutions of the test compound;
b) comparing the polarization signal generated in the absence of
the test compound with the one generated in the presence of
different concentrations of the test compound, and
c) evaluating the ability of the test compound to displace the
compound of formula (IP) as defined above indicated from a
decreased fluorescence polarization level.
Preferably, for the screening method above cited, both the PARP
protein and the 5H-phenanthridin-6-one-derived probe of formula
(IP) are pre-mixed, or the PARP protein and the test compound are
pre-mixed. In a further preferred screening method, the PARP
proteins are PARP-1, PARP-2 and PARP-3. The term "PARP protein"
encompasses full-length native proteins as well as fragments
thereof. More preferably, R.sub.11 is hydrogen or methyl, m is 0 or
1; when m is 1, n is 3 or 6, X.sup.--- is trifluoroacetate. The
5H-phenanthridin-6-one-derived probe (IP) was selected for its
capability of binding to the PARP proteins, both encompassing
full-length native proteins and fragments thereof.
The polarization signal can be measured, e.g., by a plate reader
such as the Saphire2 (Tecan). Data analysis was performed, e.g., by
using the Dynafit software. Displacement data were also fitted,
e.g., by using Excel spreadsheet (Microsoft Inc. Seattle, USA) to a
four parameter logistic model (4PL), or Hill-Slope model. The assay
was used to test compounds of the present invention. The
displacement ability of the test compounds of formula (I) is in
correlation with the compounds affinity for the NAD pocket of the
enzyme. Specific probes of formula (IP) used in the assay are: P1.
9-Dimethylamino-11,11-dimethyl-1-(3-{methyl-[(6-oxo-5,6-dihydro-phenanthr-
idin-2-ylcarbamoyl)-methyl]-carbamoyl}-propyl)-2,3,4,11-tetrahydro-naphtho-
[2,3-g]quinolinium trifluoroacetate; P2.
9-Dimethylamino-11,11-dimethyl-1-[3-(3-{[(6-oxo-5,6-dihydro-phenanthridin-
-2-ylcarbamoyl)-methyl]-amino}-propylcarbamoyl)-propyl]-2,3,4,11-tetrahydr-
o-naphtho[2,3-g]quinolinium trifluoroacetate; P3.
9-Dimethylamino-11,11-dimethyl-1-[3-(6-{[(6-oxo-5,6-dihydro-phenanthridin-
-2-ylcarbamoyl)-methyl]-amino}-hexylcarbamoyl)-propyl]-2,3,4,11-tetrahydro-
-naphtho[2,3-g]quinolinium trifluoroacetate.
A compound of formula (IP) as defined above can be prepared as
described in WO 2010/133647.
The assay is based on the use of a probe of formula (IP) that binds
to the NAD binding pocket and takes advantage of the significant
change in the polarization signal observed upon binding of the
probe to PARP-1, -2 and -3. The ability of the probe of formula
(IP) to bind full-length PARP-1, -2 and -3 has been previously
reported (WO 2010/133647). The assay has been validated as
described in WO 2010/133647.
Affinity binding constants (Kd) and DC.sub.50s (the compound
concentration at which the polarization signal is diminished by 50%
compared to untreated controls) of the test compounds can be
determined as explained in WO 2010/133647. The assay, by using
either probe P1 or probe P3, was used to evaluate the biochemical
potency of compounds of formula (I), as reported in Table 1.
TABLE-US-00001 TABLE 1 PARP-1 PARP-1 PARP-2 PARP-2 PARP-3 PARP-3
Compound (DC.sub.50 .mu.M) (Kd .mu.M) (DC.sub.50 .mu.M) (Kd .mu.M)
(DC.sub.50 .mu.M) (Kd .mu.M) (2) <0.25.dagger. <0.03.dagger.
0.34 0.18 -- -- (3) <0.25 <0.03 7.1 5.8 -- -- (4) 0.39 --
1.92 -- -- -- (5) 0.33 -- 1.64 -- -- -- (6) <0.25 0.04 7.44 5.87
-- -- (7) <0.25 0.06 0.76 0.42 -- -- (8) <0.25 0.05 >10 --
-- -- (9) <0.25 <0.03 4.94 1.44 -- -- (10) <0.25 0.07 8.4
6.8 -- -- (11) <0.25 <0.01*.dagger. 2.53 1.4 2.06 -- (12)
<0.25 <0.03 3.37 1.7 -- -- (13) <0.25 <0.03 5.08 -- --
-- (14) <0.25 <0.03 1.25 0.58 -- -- (15) <0.25 0.04 >10
-- 0.9 -- (16) <0.25 <0.03 2.88 0.98 -- -- (17) <0.25
<0.03 0.63 0.29 -- -- (19) <0.25 <0.03 2.76 1.39 -- --
(20) <0.25 <0.03 0.58 0.19 -- -- (21) 0.28 -- >10 -- -- --
(22) <0.25 <0.03 2.12 -- -- -- (23) <0.25 <0.03 2.00 --
-- -- (24) <0.25 <0.03 0.83 044 -- -- (25) <0.25 -- 5.57
-- -- -- (26) 3.33 -- >10 -- -- -- (27) 1.05 -- >10 -- -- --
(28) <0.25 <0.03 1.81 -- 5.48 -- (29) <0.25
<0.01*.dagger. 2.92 -- 1.00 -- (30) <0.25 -- >10 -- 0.35
-- (31) <0.25 -- >10 -- -- -- (32) <0.25 0.048 1.88 0.66
-- -- *Assay performed with compound P3 as the probe. In all other
cases compound P1 was used as the probe. .dagger.Assay sensitivity
limits based on a fitting error <50%.
From the above data, it is clear to a person skilled in the art
that compounds of formula (I) of the present invention are highly
potent as PARP-1 inhibitors and extremely selective versus PARP-2
and PARP-3 (compare PARP-1, PARP-2 and PARP-3 DC.sub.50 and Kd
values in Table 1 above).
Cellular Assays
PAR Assay
Cellular activity of PARP-1 inhibitors was assessed by measuring
the inhibition of the hydrogen peroxide induced PAR formation in
HeLa cells (ECACC). Cellular PAR levels were measured by
immunocytochemistry, and quantified using an ArrayScan vTi
instrument (Cellomics Thermo Scientific).
Studies were performed as follows: 6000 cells/well were seeded in
96 well plates (Perkin Elmer) in MEM/10% FCS and incubated for 24 h
at 37.degree. C., 5% carbon dioxide. Test compounds were then added
at the required concentration for 30 min. DNA damage was then
induced adding hydrogen peroxide at the concentration of 0.1 mM for
15 min. Concentration curves were prepared in MEM/10% FCS from
compound stocks in DMSO, and final DMSO concentration was 0.002%
(v/v). Duplicate wells for each concentration point were prepared
with a typical highest compound concentration of 20 .mu.M and
serial dilution 1:3. Plates were dried and fixed adding cold
methanol-acetone (70:30) solution for 15 min at room temperature,
fixing solution was aspired and wells were air dried for 5 min and
then dehydrated in PBS. Non-specific binding sites were blocked by
incubating wells for 30 min in PBS containing 5% (w/v) FBS 0.05%
tween20. Wells were then incubated for 1 h at room temperature in
PBS containing anti PAR mouse monoclonal antibody (Anti-PAR, Mouse
mAb 10H, Tulip Cat N.sup.o 1020) diluted 1:200 in blocking
solution. After 3 washes in PBS, wells incubated in PBS (w/v) 5%
FBS 0.05% Tween20 containing 2 .mu.g/mL Cy2-conjugated Goat anti
mouse secondary antibody (Amersham Pharmacia Biotech cat. N.sup.o
PA 42002) (Absorption maximum 489 nm fluorescence maximum 506 nm)
and 1 .mu.g/mL DAPI (Absorption maximum 359 nm fluorescence maximum
461 nm) (4',6-diamidino-2-phenyindole dilactate) (Sigma cat.
N.sup.o D9564), a high-sensitivity dye for nucleic acid staining.
After washing further 3 times in PBS, cellular PAR immunoreactivity
was assessed using the ArrayScan vTi instrument, with a Zeiss
10.times.0.5 N.A. objective, and applying the Cytotoxicity. V3
algorithm (Cellomics/Thermo Fisher) with a XF100 filter. At least
10 fields, corresponding to at least 900 cells, were read for each
well. IC.sub.50 Values represent the compound concentration at
which cellular PAR signal is diminished by 50% compared with
untreated controls. The following formula is used:
IC.sub.50=Bottom+(Top-Bottom)/(1+10^((Log EC.sub.50-X)));
X is the logarithm of concentration, IC.sub.50 is the response;
IC.sub.50 starts at bottom and goes to top with a sigmoid shape.
Given the above assays, compounds of formula (I) of the present
invention inhibited PAR formation with IC.sub.50 values lower than
5 .mu.M, as depicted in Table 2.
TABLE-US-00002 TABLE 2 PAR assay Compound (IC.sub.50 .mu.M) (3)
2.25 (6) 2.15 (9) 1.40 (11) 0.02 (12) 3.9 (13) 0.98 (14) 0.011 (15)
0.1 (16) 0.02 (17) 1.51 (19) 0.33 (20) 0.83 (21) 0.60 (22) 0.40
(23) 0.40 (24) 0.11 (25) 0.20 (28) 0.56 (29) 0.5 (31) 0.2 (32)
0.17
Colony Forming Assay
MDA-MB-436 breast cancer BRCA-1 mutated cells were grown at the
density of 600 cells/cm.sup.2 in RPMI medium supplemented with 10%
Fetal Bovine Serum. 24 h later different doses of compounds were
added starting from 10 .mu.M concentration in duplicates. Ten days
later, cells were fixed and stained with crystal violet. Colonies
were counted using Infrared Scanner (Odyssey Li-Cor). Anti
proliferative IC.sub.50 was calculated using Prism.
Pharmacokinetics
The pharmacokinetic profile and the oral bioavailability of the
compounds have been investigated in the mouse (Balb, Nu/Nu, Harlan,
Italy) in ad hoc pharmacokinetic studies. The compounds were
formulated in 10% tween 80/dextrose for intravenous bolus
administration while oral administrations were performed using the
compounds formulated in 0.5% methylcellulose. A single
administration at the dose of 10 mg/kg was given and three male
animals for each route were used. All blood samples were taken from
retro-orbital vein at 5 min, 30 min, 1 h, 3 h, 6 h, 24 h after
intravenous administration and 15 min, 30 min, 1 h, 3 h, 6 h, 24 h
after oral administration. Plasma samples were prepared by plasma
proteins precipitation adding 200 .mu.L of acetonitrile to 20 .mu.L
of plasma in a 96 well plate. After capping and vortex mixing, the
plate was centrifuged for 15 min at 4000 rpm. The supernatant was
considered as final extract and injected onto the LC-MS-MS system
(UPLC system: Waters Acquity using BEH C18 50*2.1 mm 1.7 .mu.m
analytical column; MS instrument: Waters TQD equipped with
Electro-Spray source operating in positive ion mode). Lower limit
of quantification is 5.0 ng/mL, upper limit of quantification is
5000 ng/mL. Non-compartmental method (linear trapezoidal rule and
linear regression analysis of natural log-transformed plasma
concentrations vs. time data) was used. Absolute bioavailability
(F) was calculated from the ratio of average oral to IV
(intravenous) dose-normalized plasma AUC (area under curve)
values.
The abbreviations used herein have the following meaning:
AUC (area under the plasma concentration vs. time curve up to the
last detectable concentration)
Cl (plasma clearance)
Cmax (maximum plasma concentration)
T1/2 (terminal half life)
Vdss (volume of distribution at steady state)
Some representative compounds of formula (I) were evaluated for
their pharmacokinetic parameters as reported in Table 3 as mean
value.
TABLE-US-00003 TABLE 3 CI (IV bolus) Vdss (IV bolus) AUC (oral)
Cmax (oral) T1/2 (oral) F on AUC Compound mL/min/kg L/Kg .mu.M h
.mu.M h % (3) 63.1 1.54 2.51 3.06 0.68 30 (11) 41.3 4.47 20.4 3.87
2.72 100 (14) 81.8 2.68 3.22 2.29 0.89 57 (15) 16.9 0.79 19.2 8.73
1.01 68 (16) 79.3 2.51 2.13 2.97 0.63 37
From the above, it is clear to the person skilled in the art that
compounds of formula (I) possess good to excellent pharmacokinetics
profiles and oral bioavailability.
In Vivo Efficacy Studies
Balb, athymic Nu/Nu male mice, from Harlan (Italy), were maintained
in agreement with the European Communities Council Directive no.
86/609/EEC concerning the protection of animals used for
experimental or other scientific purposes, in cages with paper
filter cover, food and bedding sterilized and acidified water.
Fragments of Capan-1 human pancreatic cancer tumors were implanted
subcutaneously. Mice bearing a palpable tumor (100-200 mm.sup.3)
were selected and randomized into control and treated groups. Each
group included seven animals. The treatment started one day after
randomization. Compound of formula (I) was administered by oral
route as a methocel suspension at the indicated doses and times.
Tumor dimension was measured regularly by calipers during the
experiments and tumor mass was calculated as described in Simeoni
M. et al., Cancer Res 64, 1094-1101 (2004). The tumor growth
inhibition (TGI, %) was calculated according to the equation: %
TGI=100-(mean tumor weight of treated group/mean tumor weight of
control group)*100.
Some representative compounds of formula (I) were evaluated for
their anti-tumor activity as single agent on Capan-1 BRCA-2 mutated
mouse model and results are reported in table 4. Toxicity was
evaluated on the basis of body weight reduction (no body weight
reduction observed out of 7 mice treated).
TABLE-US-00004 TABLE 4 Compounds Dose Schedule Max TGI (%) Toxicity
(11) 75 mg/kg 1-8 daily 42% 0/7 (15) 75 mg/kg 1-10 daily 54%
0/7
Representative compounds of formula (I) were evaluated for their
anti-tumor activity on Capan-1 BRCA-2 mutated mouse model in
combination with temozolomide. Compounds of formula (I) and
temozolomide were both administered by oral route. Tumor growth was
assessed by caliper. The two diameters were recorded and the tumor
weight was calculated according to the following formula: length
(mm).times.width.sup.2/2. The effect of the antitumor treatment was
evaluated as the delay in the onset of an exponential growth of the
tumor (see for references Anticancer drugs 7:437-60, 1996). This
delay (T-C value) was defined as the difference of time (in days)
required for the treatment group (T) and the control group (C)
tumors to reach a predetermined size (1 g). Toxicity was evaluated
on the basis of body weight reduction and animal survival rate. The
T-C observed when Compounds of formula (I) were combined with
temozolomide was superior to the one expected by the simple
addition of T-C obtained by the single treatments, thus indicating
strong synergism.
Therefore, the present invention provides compounds of formula (I)
useful in therapy.
Compounds of formula (I) of the present invention, suitable for
administration to a mammal, e.g., to humans, can be administered by
the usual routes and the dosage level depends upon the age, weight,
conditions of the patient and administration route.
For example, a suitable dosage adopted for oral administration of a
compound of formula (I) may range from about 1 to about 1000 mg per
dose, from 1 to 5 times daily. The compounds of the invention can
be administered in a variety of dosage forms, e.g., orally, in the
form of tablets, capsules, sugar or film coated tablets, liquid
solutions or suspensions; rectally in the form of suppositories;
parenterally, e.g., intramuscularly, or through intravenous and/or
intrathecal and/or intraspinal injection or infusion.
As stated above, the present invention also includes pharmaceutical
compositions comprising a compound of formula (I) or a
pharmaceutically acceptable salt thereof in association with a
pharmaceutically acceptable excipient, which may be a carrier or a
diluent.
The pharmaceutical compositions containing the compounds of the
invention are usually prepared following conventional methods and
are administered in a suitable pharmaceutical form. For example,
the solid oral forms may contain, together with the active
compound, diluents, e.g., lactose, dextrose, saccharose, sucrose,
cellulose, corn starch or potato starch; lubricants, e.g., silica,
talc, stearic acid, magnesium or calcium stearate, and/or
polyethylene glycols; binding agents, e.g., starches, arabic gum,
gelatine methylcellulose, carboxymethylcellulose or polyvinyl
pyrrolidone; disintegrating agents, e.g., starch, alginic acid,
alginates or sodium starch glycolate; effervescing mixtures;
dyestuffs; sweeteners; wetting agents such as lecithin,
polysorbates, laurylsulphates; and, in general, non-toxic and
pharmacologically inactive substances used in pharmaceutical
formulations. These pharmaceutical preparations may be manufactured
in known manner, for example, by means of mixing, granulating,
tabletting, sugar-coating, or film-coating processes.
The liquid dispersions for oral administration may be, e.g.,
syrups, emulsions and suspensions. As an example, the syrups may
contain, as carrier, saccharose or saccharose with glycerine and/or
mannitol and sorbitol.
The suspensions and the emulsions may contain, as examples of
carriers, natural gum, agar, sodium alginate, pectin,
methylcellulose, carboxymethylcellulose, or polyvinyl alcohol. The
suspension or solutions for intramuscular injections may contain,
together with the active compound, a pharmaceutically acceptable
carrier, e.g., sterile water, olive oil, ethyl oleate, glycols,
such as propylene glycol, and, if desired, a suitable amount of
lidocaine hydrochloride. The solutions for intravenous injections
or infusions may contain, as a carrier, sterile water or preferably
they may be in the form of sterile, aqueous, isotonic, saline
solutions or they may contain propylene glycol as a carrier.
The suppositories may contain, together with the active compound, a
pharmaceutically acceptable carrier, e.g., cocoa butter,
polyethylene glycol, a polyoxyethylene sorbitan fatty acid ester
surfactant or lecithin.
EXPERIMENTAL SECTION
For a reference to any specific compound of formula (I) of the
invention, optionally in the form of a pharmaceutically acceptable
salt, see the experimental section and claims. Referring to the
examples that follow, compounds of the present invention were
synthesized using the methods described herein, or other methods,
which are well known in the art.
The short forms and abbreviations used herein have the following
meaning:
amu (atomic mass unit)
calcd. (calculated)
.mu.M (micromolar)
.mu.L (microliter)
.mu.m (micrometer)
mol (moles)
mM (millimolar)
mmol (millimoles)
nm (nanometers)
g (grams)
mg (milligrams)
ng (nanograms)
h (hour/s)
min (minute/s)
DC.sub.50 (the half maximal Displacement Concentration)
IC.sub.50 (the half maximal Inhibitory Concentration)
PAR (poly (ADP-ribose))
MEM (Minimal Essential Medium)
FCS (Fetal Calf Serum)
FBS (Fetal Bovine Serum)
PBS (Phosphate Buffered Saline)
LC-MS (Liquid Chromatography-Mass Spectrometry)
HPLC (High Performance Liquid Chromatography)
TLC (Thin Layer Chromatography)
MHz (megahertz)
Hz (Hertz)
DMSO-d.sub.6 (deuterated dimethylsulfoxide)
CDCl.sub.3 (deuterated chloroform)
ESI (electrospray ionization)
With the aim at better illustrating the present invention, without
posing any limitation to it, the following examples are now
given.
As used herein the symbols and conventions used in the processes,
schemes and examples are consistent with those used in the
contemporary scientific literature, for example, the Journal of the
American Chemical Society or the Journal of Biological
Chemistry.
Unless otherwise noted, all materials were obtained from commercial
suppliers, of the best grade and used without further purification.
Anhydrous solvent such as N,N-dimethylformamide, tetrahydrofuran,
dichloromethane and toluene were obtained from the Aldrich Chemical
Company. All reactions involving air- or moisture-sensitive
compounds were performed under nitrogen or argon atmosphere.
General Purification and Analytical Methods
Flash Chromatography was performed on silica gel (Merck grade 9395,
60 .ANG.). HPLC was performed on Waters X Terra RP 18 (4.6.times.50
mm, 3.5 .mu.m) column using a Waters 2790 HPLC system equipped with
a 996 Waters PDA detector and Micromass mod. ZQ single quadrupole
mass spectrometer, equipped with an electrospray (ESI) ion source.
Mobile phase A was ammonium acetate 5 mM buffer (pH 5.5 with acetic
acid-acetonitrile 95:5), and mobile phase B was water-acetonitrile
(5:95). Gradient from 10 to 90% B in 8 min, hold 90% B 2 min. UV
detection at 220 nm and 254 nm. Flow rate 1 mL/min. Injection
volume 10 .mu.L. Full scan, mass range from 100 to 800 amu.
Capillary voltage was 2.5 KV; source temperature was 120.degree.
C.; cone was 10 V. Retention times (HPLC r.t.) are given in min at
220 nm or at 254 nm. Mass are given as m/z ratio.
When necessary, compounds were purified by preparative HPLC on a
Waters Symmetry C.sub.18 (19.times.50 mm, 5 .mu.m) column or on a
Waters.times.Terra RP 18 (30.times.150 mm, 5 .mu.m) column using a
Waters preparative HPLC 600 equipped with a 996 Waters PDA detector
and a Micromass mod. ZMD single quadrupole mass spectrometer,
electron spray ionization, positive mode. Mobile phase A was
water-0.01% trifluoroacetic acid, and mobile phase B was
acetonitrile. Gradient from 10 to 90% B in 8 min, hold 90% B 2 min.
Flow rate 20 mL/min. In alternative, mobile phase A was water-0.1%
ammonium hydroxide, and mobile phase B was acetonitrile. Gradient
from 10 to 100% B in 8 min, hold 100% B 2 min. Flow rate 20
mL/min.
.sup.1H-NMR spectra were performed in DMSO-d.sub.6 or CDCl.sub.3 on
a Varian Inova 400 operating at 400.5 MHz and on a Varian Mercury
300 operating at 300.0 MHz. .sup.13C NMR spectra were performed in
DMSO-d.sub.6 at 75.0 MHz.
Residual solvent signal was used as reference (.delta.=2.50 or 7.27
ppm). Chemical shifts (.delta.) are reported in parts per million
(ppm) and coupling constants (J) in Hz. The following abbreviations
are used for multiplicities: s=singlet; br. s.=broad signal;
d=doublet; t=triplet; m=multiplet; dd=doublet of doublets.
ESI(+) high-resolution mass spectra (HRMS) were obtained on a Q-Tof
Ultima (Waters, Manchester, UK) directly connected with a 1100
micro-HPLC system (Agilent, Palo Alto, US) as previously described
(Colombo, M., Sirtori, F. R., and Rizzo, V. (2004) A fully
automated method for accurate mass determination using
high-performance liquid chromatography with a quadrupole/orthogonal
acceleration time-of-flight mass spectrometer. Rapid Commun. Mass
Spectrom. 18, 511-517).
Example 1
Step a
2-Bromo-4-fluoro-6-methyl-phenylamine (X) [Hal=Br]
A solution of N-bromosuccinimide (18.7 g, 0.105 mol) in 70 mL of
N,N-dimethylformamide was added dropwise to a solution of
4-fluoro-2-methyl-phenylamine (XI) (12.5 g, 0.1 mol) in 70 mL of
the same solvent at 20.degree. C. The reaction mixture was stirred
overnight. The dark solution was poured into a mixture of water
(1000 mL), brine (50 mL) and ethyl acetate (300 mL). The mixture
was transferred into a separatory funnel, shaken and separated. The
aqueous phase was extracted with ethyl acetate (4.times.150 mL).
The combined organic layers were washed with water (5.times.100
mL), brine (2.times.100 mL), dried over Na.sub.2SO.sub.4, filtered
and concentrated. The product was purified by flash chromatography
(eluent ethyl acetate:n-hexane=1:8). The pure fractions were
combined and evaporated to give 14.9 g of product. The impure
fractions were combined, concentrated, re-dissolved in diethyl
ether (30 mL) and extracted with 5% hydrochloric acid (5.times.10
mL). The acidic phase was basified with aqueous potassium hydroxide
and extracted with diethyl ether to provide further 0.8 g of the
title compound. Total yield was 15.7 g (77%).
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 2.16 (s, 3H),
4.83 (br. s, 2H), 6.91 (dd, J.sub.H--F=9.3 Hz, J.sub.H--H=2.9 Hz,
1H), 7.16 (dd, J.sub.H--F=8.3 Hz, J.sub.H--H=2.9 Hz, 1H).
Step b
2-Bromo-4-fluoro-6-methyl-benzonitrile (IX) [Hal=Br]
A solution of potassium cyanide (16.25 g, 0.25 mol) in 20 mL of
water was added to a suspension of freshly prepared copper(I)
chloride (9.5 g, 0.096 mol) in 40 mL of water. Toluene (30 mL) was
then added and the mixture was chilled to 0.degree. C.
2-Bromo-4-fluoro-6-methyl-phenylamine (X) (15.7 g, 0.077 mol) was
added to a mixture of 16.5 ml of 36% aqueous hydrochloric acid and
40 mL of water. The resultant suspension was heated until a
solution was formed. The solution was chilled to 2.degree. C. and
the amine hydrochloride precipitated. A solution of sodium nitrite
(5.34 g, 0.078 mol) in 15 mL of water was slowly added, keeping the
reaction mixture temperature below 5.degree. C. Powdered sodium
carbonate decahydrate was added in small portions to adjust the pH
of the reaction mixture to about 7. The resultant solution of the
diazonium salt was then slowly added to the previously prepared
cyanocuprate reagent, again keeping the reaction temperature below
5.degree. C. A bright red-orange precipitate formed. The reaction
mixture was allowed to warm to 20.degree. C. and kept at this
temperature overnight. Then it was slowly heated to 70.degree. C.
for 1 h. The precipitate dissolved almost completely. The reaction
mixture was allowed to cool to 20.degree. C. and filtered. The
organic phase was separated, and the aqueous phase was extracted
with toluene (3.times.70 mL). The combined organic layers were
washed with water (2.times.100 mL), brine (2.times.100 mL), dried
over Na.sub.2SO.sub.4, filtered and concentrated. The crude nitrile
(IX) (13.9 g, 84%) was used without further purification.
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 2.52 (s, 3H),
7.44 (dd, J.sub.H--F=9.4 Hz, J.sub.H--H=2.1 Hz, 1H), 7.73 (dd,
J.sub.H--F=8.2 Hz, J.sub.H--H=2.1 Hz, 1H).
.sup.13C NMR (75.0 MHz, DMSO-d.sub.6) .delta. 115.8, 112.7 (d,
J.sub.C--F=3 Hz), 117.0 (d, J.sub.C--F=23 Hz), 118.4 (d,
J.sub.C--F=27 Hz), 126.1 (d, J.sub.C--F=11 Hz), 147.8 (d,
J.sub.C--F=11 Hz), 163.5 (d, J.sub.C--F=257 Hz).
Step c
2-Bromo-4-fluoro-6-methyl-benzamide (VIII) [Hal=Br]
2-Bromo-4-fluoro-6-methyl-benzonitrile (IX) (0.428 g, 2 mmol) was
heated in 70% aqueous sulfuric acid (2 mL) overnight at 150.degree.
C. The reaction mixture was poured into ice and extracted with
ethyl acetate (4.times.2 mL). The organic phase was washed with
water (4.times.2 mL), brine (2.times.2 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated to give 300 mg of crude
2-bromo-4-fluoro-6-methyl-benzamide (VIII). Pure sample was
obtained by recrystallization from benzene.
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 3.31 (s, 3H),
7.17 (dd, J.sub.H--F=9.8 Hz, J.sub.H--H=2.2 Hz, 1H), 7.41 (dd,
J.sub.H--F=8.6 Hz, J.sub.H--H=2.2 Hz, 1H), 7.89 (br. s, 1H), 7.65
(br. s, 1H).
Step d
2-Bromo-4-fluoro-6-methyl-benzoic Acid (VII) [Hal=Br]
2-Bromo-4-fluoro-6-methyl-benzamide (VIII) (0.9 g, 3.9 mmol) was
dissolved in 75% aqueous sulfuric acid (4 mL) at 80.degree. C.
Sodium nitrite (0.5 g, 7.2 mmol) was carefully added in small
portions during 1 h. The reaction mixture was chilled to 20.degree.
C. and cold water (15 ml) was added to the reaction mixture. The
product was extracted with ethyl acetate (6.times.2 mL). The
organic phase was washed with water (4.times.2 mL), brine
(2.times.2 mL), dried over Na.sub.2SO.sub.4, filtered and
concentrated to give 0.879 g (97%) of pure acid (VII).
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 2.31 (s, 1H),
7.22 (dd, J.sub.H--F=9.6 Hz, J.sub.H--H=2.2 Hz, 1H), 7.47 (dd,
J.sub.H--F=8.5 Hz, J.sub.H--H=2.4 Hz, 1H), 13.7 (br. s, 1H).
.sup.13C NMR (75.0 MHz, DMSO-d.sub.6+CCl.sub.4) .delta. ppm 19.5,
116.0 (d, J.sub.C--F=22 Hz), 116.8 (d, J.sub.C--F=24 Hz), 118.3 (d,
J.sub.C--F=10 Hz), 134.0 (d, J.sub.C--F=3 Hz), 138.4 (d,
J.sub.C--F=8 Hz), 163.0 (d, J.sub.C--F=250 Hz), 168.0.
Step f
2-Bromo-4-fluoro-6-methyl-benzoic Acid Methyl Ester (VI) [Hal=Br;
T=Methyl]
A mixture of 2-bromo-4-fluoro-6-methyl-benzoic acid (VII) (1.94 g,
8.33 mmol), anhydrous potassium carbonate (1.72 g, 12.5 mmol),
methyl iodide (2.36 g, 17 mmol) in N,N-dimethylformamide (15 mL)
was vigorously stirred for 23 h at 20.degree. C. The suspension was
poured into 70 mL of water. A dense oil separated out. The product
was extracted with ethyl acetate (4.times.25 mL). The organic phase
was washed with water (5.times.20 mL), brine (2.times.20 mL), dried
over Na.sub.2SO.sub.4, filtered and concentrated to give 2.07 g
(quantitative yield) of 2-bromo-4-fluoro-6-methyl-benzoic acid
methyl ester (VI).
.sup.1H NMR (400.5 MHz, CDCl.sub.3) .delta. ppm 2.35 (s, 3H), 3.96
(s, 3H), 6.91 (dd, J.sub.H--F=9.0 Hz, J.sub.H--H=2.2 Hz, 1H), 7.18
(dd, J.sub.H--F=8.1 Hz, J.sub.H--H=2.4 Hz, 1H).
Step g
2-Cyano-4-fluoro-6-methyl-benzoic Acid Methyl Ester (V)
[T=Methyl]
A mixture of 2-bromo-4-fluoro-6-methyl-benzoic acid methyl ester
(VI) (275 mg, 1.12 mmol), potassium hexacyanoferrate (II) (206 mg,
0.56 mmol), anhydrous sodium carbonate (237 mg, 2.24 mmol) and
palladium(II) acetate (5 mg, 0.0224 mmol) in 3 mL of
N-methylpyrrolidone was heated at 120.degree. C. in a sealed tube
under argon atmosphere overnight. The reaction mixture was diluted
with dichloromethane and filtered through a pad of Celite. The
organic phase was washed with water (13.times.6 mL), brine
(2.times.6 mL), dried over Na.sub.2SO.sub.4, filtered and
concentrated. Column chromatography (n-hexane/ethyl acetate:7/3)
afforded 2-cyano-4-fluoro-6-methyl-benzoic acid methyl ester (76
mg, 35%).
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 2.42 (s, 3H),
3.93 (s, 3H), 7.65 (dd, J.sub.HF=9.6, J.sub.HH=2.6 Hz, 1H), 7.85
(dd, J.sub.HF=8.3, 2.6 Hz, 1H).
Step h
6-Fluoro-2-(3-morpholin-4-yl-propyl)-3-oxo-2,3-dihydro-1H-isoindole-4-carb-
onitrile (IV) [n=3; R1=morpholin-4-yl; X=Null, as m=0 and
R2=Null]
To a solution of 2-cyano-4-fluoro-6-methyl-benzoic acid methyl
ester (V) (208 mg, 1.07 mmol) in methyl pivalate (2 mL),
N-bromosuccinimide (310 mg, 1.74 mmol) and benzoylperoxide (20 mg,
0.097 mmol) were added. The reaction mixture was stirred at
85.degree. C. under nitrogen atmosphere for 3 h. Crude was filtered
on Gooch and washed with toluene. Volatiles were evaporated and the
residue was dissolved in acetonitrile (3 mL). Triethylamine (0.41
mL, 2.9 mmol) and 3-morpholin-4-yl-propylamine (XIII) (140 mg, 0.97
mmol) were added and the reaction mixture was stirred at 90.degree.
C. for 3 h. Crude was diluted with dichloromethane and washed with
15% ammonium hydroxide. The organic phase was dried over
Na.sub.2SO.sub.4, filtered and evaporated. Column chromatography
(gradient from chloroform/methanol: 96/4 to chloroform/methanol:
94/6) afforded
6-fluoro-2-(3-morpholin-4-yl-propyl)-3-oxo-2,3-dihydro-1H-isoind-
ole-4-carbonitrile (IV) (130 mg, 40% yield).
.sup.1H NMR (400.5 MHz, CDCl.sub.3) .delta. ppm 1.87 (quintet,
J=7.1 Hz, 2H), 2.34-2.49 (m, 6H), 3.62-3.74 (m, 6H), 4.45 (s, 2H),
7.42 (dd, J.sub.H--F=7.3 Hz, J.sub.H--H=2.0 Hz, 1H), 7.47 (dd,
J.sub.H--F=8.3 Hz, J.sub.H--H=2.0 Hz, 1H).
Step c'
6-Fluoro-2-(3-morpholin-4-yl-propyl)-3-oxo-2,3-dihydro-1H-isoindole-4-carb-
oxylic Acid Amide (I), cpd 23
[R=F; n=3; R1=morpholin-4-yl; m=0; R2=null]
##STR00023##
A solution of
6-fluoro-2-(3-morpholin-4-yl-propyl)-3-oxo-2,3-dihydro-1H-isoindole-4-car-
bonitrile (IV) (100 mg, 0.33 mmol) in 1.5 mL of 36% hydrochloric
acid was heated at 50.degree. C. for 10 h. All volatile materials
were evaporated and the residue was dissolved in 2 mL of cold
water. The solution was neutralized with solid potassium carbonate.
The solid precipitated was dissolved in dichloromethane and the
organic phase was washed with saturated aqueous sodium carbonate
(2.times.1 mL), brine (2.times.1 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated to give 73 mg (73%) of
6-fluoro-2-(3-morpholin-4-yl-propyl)-3-oxo-2,3-dihydro-1H-isoind-
ole-4-carboxylic acid amide (I).
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.79 (quintet,
J=7.1 Hz, 2H), 2.28-2.35 (m, 6H), 3.47-3.52 (m, 4H), 3.59 (t, J=7.1
Hz, 2H), 4.58 (s, 2H), 7.68 (dd, J.sub.HF=7.8, J.sub.HH=2.6 Hz,
1H), 7.83 (br. s., 1H), 7.89 (dd, J.sub.HF=10.9, J.sub.HH=2.6 Hz,
1H), 10.81 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.21FN.sub.3O.sub.3 [M+H].sup.+
322.1562. found 322.1565.
Example 2
Step e
4-Fluoro-2-iodo-6-methyl-benzoic Acid (VII) [Hal=I]
A mixture of 4-fluoro-2-methyl-benzoic acid (XII) (20.00 g, 0.130
mol), iodobenzene diacetate (50.15 g, 0.156 mol), iodine (39.52 g,
0.156 mol) and palladium(II) acetate (1.46 g, 0.006 mol) in
N,N-dimethylformamide (360 mL) was degassed by cycling vacuum and
nitrogen three times and then was heated for 18 h at 100.degree. C.
internal temperature, under argon. The resultant dark mixture was
cooled to room temperature, diluted with methyl-tert-butylether
(200 mL) and treated with a solution of sodium metabisulfite (250
g) in water (500 mL) under efficient stirring. Then, this yellow
colored mixture was acidified by slowly adding conc. hydrochloric
acid (130 mL). The aqueous layer was separated and extracted twice
with methyl-tert-butylether (mL100.times.2). The combined organic
extracts were treated with a solution of sodium hydroxide pellets
(80 g) in water (300 mL) under stirring. The organic layer
containing only iodobenzene was discharged, while the aqueous layer
was added with sodium chloride, cooled to ice temperature and
brought to very low pH with conc. hydrochloric acid (130 mL). From
this aqueous medium the product was extracted with
methyl-tert-butylether (100 mL.times.3) and the combined extracts
were dried over Na.sub.2SO.sub.4 and finally concentrated under
reduced pressure affording 30.5 g (84%) of
4-fluoro-2-iodo-6-methyl-benzoic acid as brown solid. This raw
material was used in the next step without purification.
.sup.1H NMR (300.0 MHz, CDCl.sub.3) .delta. ppm 2.46 (s, 3H), 6.96
(dd, J.sub.HF=9.1, J.sub.HH=2.6 Hz, 1H), 7.45 (dd, J.sub.HF=7.9,
2.3 Hz, 1H).
Step f
4-Fluoro-2-iodo-6-methyl-benzoic Acid Methyl Ester (VI) [Hal=I;
T=Methyl]
To a solution of 4-fluoro-2-iodo-6-methyl-benzoic acid (VII) (30.05
g, 0.109 mol) in N,N-dimethylformamide (300 mL) was added anhydrous
potassium carbonate (22.0 g, 0.16 mol) under efficient magnetic
stirring. After 15 min methyl p-toluensulfonate (30.7 g, 0.16 mol)
was added. The brown suspension was stirred at room temperature for
2 h. Potassium acetate (12.4 g, 0.13 mol) was then added to destroy
the unreacted methyl p-toluensulfonate and the mixture was stirred
overnight. The thick reaction mixture was diluted with
methyl-tert-butylether (100 mL) and washed with water (600 mL); the
aqueous layer was separated and extracted twice with
methyl-tert-butylether (70 mL.times.2). The combined organic
extracts were washed with brine (50 mL), dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to a solid
residue. This material was purified by chromatography (eluant
n-hexane/ethyl acetate 9:1), affording 26.2 g (81%) of product as
colorless oil.
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 2.27 (s, 3H),
3.86 (s, 3H), 7.25 (dd, J.sub.HF=9.6, J.sub.HH=2.4 Hz, 1H), 7.63
(dd, J.sub.HF=8.2, J.sub.HH=2.4 Hz, 1H).
Step g
2-Cyano-4-fluoro-6-methyl-benzoic Acid Methyl Ester (V)
[T=Methyl]
A solution of 4-fluoro-2-iodo-6-methyl-benzoic acid methyl ester
(VI) (26.02 g, 88.48 mmol) in 260 mL of N,N-dimethylformamide was
treated with copper(I) cyanide (12.18 g; 0.136 mol) and stirred at
110.degree. C. for 5 h. The dark colored mixture was allowed to
cool to about 60.degree. C., treated with 105 g of Celite.RTM. 560
coarse (Fluke) under efficient stirring and diluted with ethyl
acetate (250 mL). After cooling to room temperature, the mixture
was slowly poured in 0.25N aqueous sodium hydroxide (500 mL) and
then filtered. The reaction flask and the panel were washed with
ethyl acetate (100 mL). The aqueous layer was separated and
extracted twice with ethyl acetate (250 mL+100 mL). The combined
organic extracts were washed with brine (200 mL), dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to give
22.00 g of raw product as yellow solid. This material was
crystallized from n-hexane (40 mL): after cooling to room
temperature the solid was collected by filtration and the mother
liquors were concentrated under reduced pressure. The solid residue
so obtained was crystallized from n-hexane (20 mL) yielding, after
filtration of the solids, a second crop of product. The combined
crops (14.15 g) were finally purified by chromatography eluting in
gradient from n-hexane/methyl-tert-butylether 9:1 to n-hexane/ethyl
acetate 9:1. After evaporation of the fractions 12.0 g (70%) of
2-cyano-4-fluoro-6-methyl-benzoic acid methyl ester (V) were
obtained.
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 2.42 (s, 3H),
3.93 (s, 3H), 7.65 (dd, J.sub.HF=9.6, J.sub.HH=2.6 Hz, 1H), 7.85
(dd, J.sub.HF=8.3, 2.6 Hz, 1H).
Step h
2-(1-Cyclohexyl-piperidin-4-yl)-6-fluoro-3-oxo-2,3-dihydro-1H-isoindole-4--
carbonitrile (IV) [R=F; n=m=0; R1=piperidin-4-yl;
R2=1-cyclohexyl]
To a solution of 2-cyano-4-fluoro-6-methyl-benzoic acid methyl
ester (V) (208 mg, 1.07 mmol) in methyl pivalate (2 mL),
N-bromosuccinimide (310 mg, 1.74 mmol) and benzoylperoxide (20 mg,
0.097 mmol) were added. The reaction mixture was stirred at
85.degree. C. under nitrogen atmosphere for 3 h. Crude was filtered
and washed with toluene. Volatiles were evaporated and the residue
was dissolved in acetonitrile (3 mL). Potassium carbonate (670 mg,
4.85 mmol) and 1-cyclohexyl-piperidin-4-ylamine dihydrochloride
monohydrate (XIII) (265 mg, 0.97 mmol) were added and the reaction
mixture was stirred at 90.degree. C. for 3 h. Crude was diluted
with dichloromethane and washed with 15% ammonium hydroxide. The
organic phase was dried over Na.sub.2SO.sub.4, filtered and
evaporated. Column chromatography (dichloromethane/methanol/ammonia
solution, 7N in methanol: 97/2/1) afforded
2-(1-cyclohexyl-piperidin-4-yl)-6-fluoro-3-oxo-2,3-dihydro-1H-is-
oindole-4-carbonitrile (IV) (100 mg, 30%).
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.02-1.13 (m,
1H), 1.16-1.27 (m, 4H), 1.55-1.62 (m, 1H), 1.68-1.80 (br. s., 7H),
2.23-2.39 (m, 3H), 2.87-2.97 (m, 2H), 3.95 (br. s., 1H), 4.52 (s,
2H), 7.86 (dd, J.sub.HF=8.3, J.sub.HH=2.2 Hz, 1H), 7.98 (dd,
J.sub.HF=9.3, J.sub.HH=2.2 Hz, 1H).
HRMS (ESI+): calcd. for C.sub.20H.sub.25FN.sub.3O [M+H].sup.+
342.1976. found 342.1988.
Step c'
2-(1-Cyclohexyl-piperidin-4-yl)-6-fluoro-3-oxo-2,3-dihydro-1H-isoindole-4--
carboxylic Acid Amide (I), cpd 29
[R=F; n=m=0; R1=piperidin-4-yl; R2=1-cyclohexyl]
##STR00024##
To a stirred solution of
2-(1-cyclohexyl-piperidin-4-yl)-6-fluoro-3-oxo-2,3-dihydro-1H-isoindole-4-
-carbonitrile (IV) (100 mg, 0.3 mmol) in acetic acid (5 mL),
concentrated sulfuric acid (2.7 mL) was added dropwise during 30
min. The reaction was then warmed at 80.degree. C. for 9 h, cooled
at room temperature and poured into cold water (10 mL). The aqueous
phase was then made basic by adding concentrated aqueous ammonia
and extracted with dichloromethane (3.times.10 mL). The combined
organic phases were washed with 2N aqueous sodium hydroxide
(2.times.12 mL) and brine, dried over Na.sub.2SO.sub.4 and
evaporated to dryness in vacuo. The title compound was obtained as
a white solid (43 mg, 40%) after purification through column
chromatography ((dichloromethane/methanol/ammonia solution, 7N in
methanol: 97/2/1).
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.00-1.14 (m,
1H), 1.14-1.28 (m, 4H), 1.53-1.61 (m, 1H), 1.67-1.80 (m, 6H),
2.25-2.36 (m, 3H), 2.88-2.95 (m, 2H), 3.94-4.03 (m, 1H), 4.55 (s,
2H), 7.66 (dd, J.sub.HF=7.7, J.sub.HH=2.6 Hz, 1H), 7.85 (br. s.,
1H), 7.89 (dd, J.sub.HF=10.9, J.sub.HH=2.6 Hz, 1H), 10.78 (br. s.,
1H).
HRMS (ESI+): calcd. for C.sub.20H.sub.27FN.sub.3O.sub.2 [M+H].sup.+
360.2082. found 360.2098.
Example 3
Step m
4-[(Furan-2-ylmethyl)-amino]-piperidine-1-carboxylic acid
tert-butyl Ester (XVI) [n=0; R1=piperidin-4-yl;
X=tert-butoxycarbonyl]
To an equimolar solution of furan-2-carbaldehyde (XV) (250 mg, 2.6
mmol) and 4-amino-piperidine-1-carboxylic acid tert-butyl ester
(XIII) (473 mg, 2.6 mmol) in dichloromethane (14 mL) 1M
titanium(IV) chloride in dichloromethane (1.3 mL, 1.3 mmol) and
triethylamine (0.32 mL, 2.6 mmol) were added. The reaction mixture
was stirred under nitrogen atmosphere for 2 days. Then sodium
cyanoborohydride (493 mg, 7.8 mmol) in methanol (7 mL) was added
dropwise with stirring and the solution was allowed to stir
overnight at room temperature. 35% sodium hydroxide was added and
the product was extracted with ethyl acetate. The organic phase was
separated, washed with brine, dried over Na.sub.2SO.sub.4 and
evaporated to dryness in vacuo. The crude was purified by flash
chromatography (dichloromethane/methanol 95:5) to give the title
compound as a red oil (406 mg, 56%).
HRMS (ESI+): calcd. for C.sub.15H.sub.25N.sub.2O.sub.3 [M+H].sup.+
281.1860. found 281.1867.
Operating in an analogous way, but employing suitably substituted
starting material (XIII), the following compounds were
obtained:
Benzyl-furan-2-ylmethyl-amine (XVI)
HRMS (ESI+): calcd. for C.sub.12H.sub.14NO [M+H].sup.+ 188.1070.
found 188.1075.
Furan-2-ylmethyl-phenethyl-amine (XVI)
HRMS (ESI+): calcd. for C.sub.13H.sub.16NO [M+H].sup.+ 202.1226.
found 202.1230.
[2-(3,4-Dihydro-1H-isoquinolin-2-yl)-ethyl]-furan-2-ylmethyl-amine
(XVI)
HRMS (ESI+): calcd. for C.sub.16H.sub.21N.sub.2O [M+H].sup.+
257.1648. found 257.1642.
Furan-2-ylmethyl-(2-piperidin-1-yl-ethyl)-amine (XVI)
HRMS (ESI+): calcd. for C.sub.12H.sub.21N.sub.2O [M+H].sup.+
209.1648. found 209.1650.
Furan-2-ylmethyl-(2-morpholin-4-yl-ethyl)-amine (XVI)
HRMS (ESI+): calcd. for C.sub.11H.sub.19N.sub.2O.sub.2 [M+H].sup.+
211.1441. found 211.1446.
Furan-2-ylmethyl-(3-morpholin-4-yl-propyl)-amine (XVI)
HRMS (ESI+): calcd. for C.sub.12H.sub.21N.sub.2O.sub.2 [M+H].sup.+
225.1598. found 225.1590.
[2-(3,4-Dihydro-2H-quinolin-1-yl)-ethyl]-furan-2-ylmethyl-amine
(XVI)
HRMS (ESI+): calcd. for C.sub.16H.sub.21N.sub.2O [M+H].sup.+
257.1648. found 257.1652.
Furan-2-ylmethyl-(3-phenyl-propyl)-amine (XVI)
HRMS (ESI+): calcd. for C.sub.14H.sub.18NO [M+H].sup.+ 216.1383.
found 216.1387.
Furan-2-ylmethyl-(2-pyridin-2-yl-ethyl)-amine (XVI)
HRMS (ESI+): calcd. for C.sub.12H.sub.15N.sub.2O [M+H].sup.+
203.1179. found 203.1181.
[3-(3,4-Dihydro-1H-isoquinolin-2-yl)-propyl]-furan-2-ylmethyl-amine
(XVI)
HRMS (ESI+): calcd. for C.sub.17H.sub.23N.sub.2O [M+H].sup.+
271.1805. found 271.1799.
[3-(3,4-Dihydro-2H-quinolin-1-yl)-propyl]-furan-2-ylmethyl-amine
(XVI)
HRMS (ESI+): calcd. for C.sub.17H.sub.23N.sub.2O [M+H].sup.+
271.1805. found 271.1811.
Furan-2-ylmethyl-[3-(4-methyl-piperazin-1-yl)-propyl]-amine
(XVI)
HRMS (ESI+): calcd. for C.sub.13H.sub.24N.sub.3O [M+H].sup.+
238.1914. found 238.1912.
Furan-2-ylmethyl-[3-(4-phenyl-piperazin-1-yl)-propyl]-amine
(XVI)
HRMS (ESI+): calcd. for C.sub.18H.sub.26N.sub.3O [M+H].sup.+
300.2070. found 300.2077.
Furan-2-ylmethyl-(3-piperidin-1-yl-propyl)-amine (XVI)
HRMS (ESI+): calcd. for C.sub.13H.sub.23N.sub.2O [M+H].sup.+
223.1805. found 223.1802.
(3-[1,4]Bipiperidinyl-1-yl-propyl)-furan-2-ylmethyl-amine (XVI)
HRMS (ESI+): calcd. for C.sub.18H.sub.32N.sub.3O [M+H].sup.+
306.2540. found 306.2544.
[3-(2,6-Dimethyl-piperidin-1-yl)-propyl]-furan-2-ylmethyl-amine
(XVI)
HRMS (ESI+): calcd. for C.sub.15H.sub.27N.sub.2O [M+H].sup.+
251.2118. found 251.2120.
Furan-2-ylmethyl-[1-(tetrahydro-pyran-4-yl)-piperidin-4-yl]-amine
(XVI)
HRMS (ESI+): calcd. for C.sub.15H.sub.25N.sub.2O.sub.2 [M+H].sup.+
265.1911. found 265.1919.
(1-Benzyl-piperidin-4-yl)-furan-2-ylmethyl-amine (XVI)
HRMS (ESI+): calcd. for C.sub.17H.sub.23N.sub.2O [M+H].sup.+
271.1805. found 271.1807.
[2-(1-Benzyl-piperidin-4-yl)-ethyl]-furan-2-ylmethyl-amine
(XVI)
HRMS (ESI+): calcd. for C.sub.19H.sub.27N.sub.2O [M+H].sup.+
299.2118. found 299.21222.
[3-(4-Benzyl-piperidin-1-yl)-propyl]-furan-2-ylmethyl-amine
(XVI)
HRMS (ESI+): calcd. for C.sub.20H.sub.29N.sub.2O [M+H].sup.+
313.2274. found 313.2280.
(1-Cyclohexyl-piperidin-4-yl)-furan-2-ylmethyl-amine (XVI)
An equimolar solution of furan-2-carbaldehyde (XV) (1.3 g, 13.5
mmol) and 1-cyclohexyl-piperidin-4-ylamine (XIII) (2.46 g, 13.5
mmol) in toluene (140 mL) was heated to reflux for 8 h by employing
a Dean-Stark apparatus. The reaction mixture was concentrated under
vacuum and rinsed with ethanol (50 mL). Sodium
triacetoxyborohydride (3.8 g, 17.93 mmol) was added and the mixture
was left overnight at room temperature. Then it was basified with
aqueous ammonia (8%) and the aqueous layer was separated and
extracted with diethyl ether. The organic phase was dried over
anhydrous sodium sulfate and concentrated under reduced pressure to
give the title compound as a yellow oil employed in the following
step without any further purification.
HRMS (ESI+): calcd. for C.sub.16H.sub.27N.sub.2O [M+H].sup.+
263.2118. found 263.2120.
Step n
3-(1-Tert-butoxycarbonyl-piperidin-4-yl)-4-oxo-10-oxa-3-aza-tricyclo[5.2.1-
.0*1,5*]dec-8-ene-6-carboxylic Acid (XVII) [n=0; R1=piperidin-4-yl;
X=tert-butoxycarbonyl]
To a solution of
4-[(furan-2-ylmethyl)-amino]-piperidine-1-carboxylic acid
tert-butyl ester (XVI) (5.6 g, 21 mmol) in toluene (300 mL) maleic
anhydride (2.1 g, 21 mmol) was added. The reaction mixture was
refluxed for 6 h and stirred overnight at room temperature. The
precipitate solid obtained was filtered, washed with diethyl ether
and dried to give the desired compound (6.5 g, 82%) as a white
solid.
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.40 (s, 9H),
1.40-1.63 (m, 4H), 2.45 (d, J=9.3 Hz, 1H), 2.75 (br. s., 2H), 2.76
(d, J=9.3 Hz, 1H), 3.59 (d, J=11.6 Hz, 1H), 3.88 (d, J=11.60 Hz,
1H), 3.90 (m, 1H), 3.96-4.06 (m, 2H), 4.95 (d, J=1.6 Hz, 1H), 6.42
(dd, J=5.6, 1.7 Hz, 1H), 6.55 (d, J=5.6 Hz, 1H), 12.03 (br. s.,
1H).
HRMS (ESI+): calcd. for C.sub.19H.sub.27N.sub.2O.sub.6 [M+H].sup.+
379.1864. found 379.1876.
Operating in an analogous way, but employing suitably substituted
starting material (XVI) the following compounds were obtained:
3-Benzyl-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]dec-8-ene-6-carboxylic
Acid (XVII)
HRMS (ESI+): calcd. for C.sub.16H.sub.16NO.sub.4 [M+H].sup.+
286.1074. found 286.1078.
4-Oxo-3-phenethyl-10-oxa-3-aza-tricyclo[5.2.1.0*1,5]*dec-8-ene-6-carboxyli-
c Acid (XVII)
HRMS (ESI+): calcd. for C.sub.17H.sub.18NO.sub.4 [M+H].sup.+
300.1230. found 300.1237.
3-[2-(3,4-Dihydro-1H-isoquinolin-2-yl)-ethyl]-4-oxo-10-oxa-3-aza-tricyclo[-
5.2.1.0*1,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.20H.sub.23N.sub.2O.sub.4 [M+H].sup.+
355.1652. found 355.1657.
4-Oxo-3-(2-piperidin-1-yl-ethyl)-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]dec-8--
ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.16H.sub.23N.sub.2O.sub.4 [M+H].sup.+
307.1652. found 307.1660.
3-(2-Morpholin-4-yl-ethyl)-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]dec-8--
ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.16H.sub.21N.sub.2O.sub.6 [M+H].sup.+
309.1445. found 309.1446.
3-(3-Morpholin-4-yl-propyl)-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]dec-8-
-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.16H.sub.23N.sub.2O.sub.6 [M+H].sup.+
323.1601. found 323.1609.
3-[2-(3,4-Dihydro-2H-quinolin-1-yl)-ethyl]-4-oxo-10-oxa-3-aza-tricyclo[5.2-
.1.0*1,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.20H.sub.23N.sub.2O.sub.4 [M+H].sup.+
355.1652. found 355.1660.
4-Oxo-3-(3-phenyl-propyl)-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]dec-8-ene-6-c-
arboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.18H.sub.20NO.sub.4 [M+H].sup.+
314.1387. found 314.1392.
4-Oxo-3-(2-pyridin-2-yl-ethyl)-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]dec-8-en-
e-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.16H.sub.17N.sub.2O.sub.4 [M+H].sup.+
301.1183. found 301.1179.
3-[3-(3,4-Dihydro-1H-isoquinolin-2-yl)-propyl]-4-oxo-10-oxa-3-aza-tricyclo-
[5.2.1.0*1,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.21H.sub.26N.sub.2O.sub.4 [M+H].sup.+
369.1809. found 369.1811.
3-[3-(3,4-Dihydro-2H-quinolin-1-yl)-propyl]-4-oxo-10-oxa-3-aza-tricyclo[5.-
2.1.0*1,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.21H.sub.25N.sub.2O.sub.4 [M+H].sup.+
369.1809. found 369.1801.
3-[3-(4-Methyl-piperazin-1-yl)-propyl]-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0-
*1,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.17H.sub.26N.sub.3O.sub.4 [M+H].sup.+
336.1918. found 336.1920.
4-Oxo-3-[3-(4-phenyl-piperazin-1-yl)-propyl]-10-oxa-3-aza-tricyclo[5.2.1.0-
*1,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.22H.sub.28N.sub.3O.sub.4 [M+H].sup.+
398.2074. found 398.2079.
4-Oxo-3-(3-piperidin-1-yl-propyl)-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]dec-8-
-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.17H.sub.26N.sub.2O.sub.4 [M+H].sup.+
321.1809. found 321.1812.
3-(3-[1,4]Bipiperidinyl-1-yl-propyl)-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0*1-
,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.22H.sub.34N.sub.3O.sub.4 [M+H].sup.+
404.2544. found 404.2540.
3-[3-(2,6-Dimethyl-piperidin-1-yl)-propyl]-4-oxo-10-oxa-3-aza-tricyclo[5.2-
.1.0*1,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.16H.sub.26N.sub.2O.sub.4 [M+H].sup.+
349.2122. found 349.2119.
4-Oxo-3-[1-(tetrahydro-pyran-4-yl)-piperidin-4-yl]-10-oxa-3-aza-tricyclo[5-
.2.1.0*1,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.16H.sub.27N.sub.2O.sub.6 [M+H].sup.+
363.1914. found 363.1920.
3-(1-Benzyl-piperidin-4-yl)-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]dec-8-
-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.21H.sub.25N.sub.2O.sub.4 [M+H].sup.+
369.1809. found 369.1799.
3-[2-(1-Benzyl-piperidin-4-yl)-ethyl]-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0*-
1,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.23H.sub.29N.sub.2O.sub.4 [M+H].sup.+
397.2122. found 397.2127.
3-[3-(4-Benzyl-piperidin-1-yl)-propyl]-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0-
*1,5*]dec-8-ene-6-carboxylic Acid (XVII)
HRMS (ESI+): calcd. for C.sub.24H.sub.31N.sub.2O.sub.4 [M+H].sup.+
411.2278. found 411.2283.
3-(1-Cyclohexyl-piperidin-4-yl)-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]d-
ec-8-ene-6-carboxylic Acid (XVII)
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 11.98 (br.s.,
1H), 6.58 (d, J=5.6 Hz, 1H), 6.44 (dd, J=5.6, 1.6 Hz, 1H), 4.97 (d,
J=1.6 Hz, 1H), 4.00-4.11 (m, 1H), 3.96 (d, J=11.0 Hz, 1H), 3.55 (d,
J=11.0 Hz 1H), 3.38-3.48 (m, 2H), 3.04-3.2 (m, 3H), 2.79 (d, J=9.1
Hz, 1H), 2.48 (d, J=9.1 Hz, 1H), 1.55-2.01 (m, 8H), 1.04-1.44 (m,
6H).
HRMS (ESI+): calcd. for C.sub.20H.sub.29N.sub.2O.sub.4 [M+H].sup.+
361.2122. found 361.2129.
Step o
3-Oxo-2-piperidin-4-yl-2,3-dihydro-1H-isoindole-4-carboxylic Acid
Hydrochloride (XIX) [n=0; R1=piperidin-4-yl]
3-(1-Tert-butoxycarbonyl-piperidin-4-yl)-4-oxo-10-oxa-3-aza-tricyclo[5.2.1-
.0*1,5*]dec-8-ene-6-carboxylic Acid (XVII)
(6.35 g, 16.8 mmol) was dissolved in 37% hydrochloric acid (80 mL)
and the resulted solution was refluxed for 3 h. The solvent was
removed under reduced pressure and the residue was diluted with
methanol and decanted to obtain the desired product (XIX) as a
white solid (4.06 g, 82%).
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.95-2.12 (m,
4H), 3.01-3.18 (m, 2H), 3.36-3.45 (m, 2H), 4.36-4.46 (m, 1H), 4.72
(s, 2H), 7.85 (dd, J=7.7, 7.5 Hz, 1H), 7.95 (dd, J=7.5, 0.8 Hz,
1H), 8.17 (dd, J=7.7, 0.8 Hz, 1H), 8.53 (br. s., 1H), 8.79 (br. s.,
1H), 15.86 (s, 1H).
HRMS (ESI+): calcd. for C.sub.14H.sub.17N.sub.2O.sub.3 [M+H].sup.+
261.1234. found 261.1222.
Operating in an analogous way, but employing suitably substituted
starting material (XVII) the following compounds were obtained:
2-Benzyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic Acid
(XVIII)
HRMS (ESI+): calcd. for C.sub.16H.sub.14NO.sub.3 [M+H].sup.+
268.0968. found 268.0972.
3-Oxo-2-phenethyl-2,3-dihydro-1H-isoindole-4-carboxylic Acid
(XVIII)
HRMS (ESI+): calcd. for C.sub.17H.sub.16NO.sub.3 [M+H].sup.+
282.1125. found 282.1131.
2-[2-(3,4-Dihydro-1H-isoquinolin-2-yl)-ethyl]-3-oxo-2,3-dihydro-1H-isoindo-
le-4-carboxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.20H.sub.21N.sub.2O.sub.3 [M+H].sup.+
337.1547. found 337.1541.
3-Oxo-2-(2-piperidin-1-yl-ethyl)-2,3-dihydro-1H-isoindole-4-carboxylic
Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.16H.sub.21N.sub.2O.sub.3 [M+H].sup.+
288.1547. found 288.1552.
2-(2-Morpholin-4-yl-ethyl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic
Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.15H.sub.19N.sub.2O.sub.4 [M+H].sup.+
291.1339. found 291.1335.
2-(3-Morpholin-4-yl-propyl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic
Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.16H.sub.21N.sub.2O.sub.4 [M+H].sup.+
305.1496. found 305.1492.
2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-ethyl]-3-oxo-2,3-dihydro-1H-isoindole--
4-carboxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.20H.sub.21N.sub.2O.sub.3 [M+H].sup.+
337.1547. found 337.1549.
3-Oxo-2-(3-phenyl-propyl)-2,3-dihydro-1H-isoindole-4-carboxylic
Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.18H.sub.18NO.sub.3 [M+H].sup.+
296.1281. found 296.1290.
3-Oxo-2-(2-pyridin-2-yl-ethyl)-2,3-dihydro-1H-isoindole-4-carboxylic
Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.16H.sub.15N.sub.2O.sub.3 [M+H].sup.+
283.1077. found 283.1080.
2-[3-(3,4-Dihydro-1H-isoquinolin-2-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoind-
ole-4-carboxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.21H.sub.23N.sub.2O.sub.3 [M+H].sup.+
351.1703. found 351.1706.
2-[3-(3,4-Dihydro-2H-quinolin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole-
-4-carboxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.21H.sub.23N.sub.2O.sub.3 [M+H].sup.+
351.1703. found 351.1699.
2-[3-(4-Methyl-piperazin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole-4-ca-
rboxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.17H.sub.24N.sub.3O.sub.3 [M+H].sup.+
318.1812. found 318.1820.
3-Oxo-2-[3-(4-phenyl-piperazin-1-yl)-propyl]-2,3-dihydro-1H-isoindole-4-ca-
rboxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.22H.sub.26N.sub.3O.sub.3 [M+H].sup.+
380.1969. found 380.1971.
3-Oxo-2-(3-piperidin-1-yl-propyl)-2,3-dihydro-1H-isoindole-4-carboxylic
Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.17H.sub.23N.sub.2O.sub.3 [M+H].sup.+
303.1703. found 303.1702.
2-(3-[1,4]Bipiperidinyl-1-yl-propyl)-3-oxo-2,3-dihydro-1H-isoindole-4-carb-
oxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.22H.sub.32N.sub.3O.sub.3 [M+H].sup.+
386.2438. found 386.2442.
2-[3-(2,6-Dimethyl-piperidin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole--
4-carboxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.16H.sub.27N.sub.2O.sub.3 [M+H].sup.+
331.2016. found 331.2011.
3-Oxo-2-[1-(tetrahydro-pyran-4-yl)-piperidin-4-yl]-2,3-dihydro-1H-isoindol-
e-4-carboxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.16H.sub.26N.sub.2O.sub.4 [M+H].sup.+
345.1809. found 345.1816.
2-(1-Benzyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic
Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.21H.sub.23N.sub.2O.sub.3 [M+H].sup.+
351.1703. found 351.1708.
2-[2-(1-Benzyl-piperidin-4-yl)-ethyl]-3-oxo-2,3-dihydro-1H-isoindole-4-car-
boxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.23H.sub.27N.sub.2O.sub.3 [M+H].sup.+
379.2016. found 379.2020.
2-[3-(4-Benzyl-piperidin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole-4-ca-
rboxylic Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.24H.sub.29N.sub.2O.sub.3 [M+H].sup.+
393.2173. found 393.2177.
2-(1-Cyclohexyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxyli-
c Acid (XVIII)
HRMS (ESI+): calcd. for C.sub.20H.sub.27N.sub.2O.sub.3 [M+H].sup.+
343.2016. found 343.2019.
Step q
2-(1-Tert-butoxycarbonyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4--
carboxylic Acid (XVIII) [n=0; R1=piperidin-4-yl;
X=tert-butoxycarbonyl]
To a solution of
3-oxo-2-piperidin-4-yl-2,3-dihydro-1H-isoindole-4-carboxylic acid
(3.9 g, 13.2 mmol) in pyridine (15 mL) potassium carbonate (3.6 g,
26.5 mmol) and methanol (40 mL) were successively added. Then
di-tert-butyl dicarbonate (3.16 g, 14.5 mmol) was added and the
reaction mixture was stirred at room temperature for 4 h until HPLC
analysis revealed the disappearance of the starting material. The
solvent was removed under reduced pressure and the residue was
dissolved in dichloromethane. The solution was washed twice with 5%
potassium hydrogen sulfate and the organic phase was dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The obtained crude wad
diluted with diethyl ether and decanted to obtain the title
compound (3.7 g, 78%) as a white solid.
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.42 (s, 9H),
1.04-1.74 (m, 2H), 1.80-1.88 (m, 2H), 2.89 (br. s., 2H), 4.04-4.12
(m, 2H), 4.23-4.32 (m, 1H), 4.73 (s, 2H), 7.83 (dd, J=7.5, 0.8 Hz,
1H), 7.91 (dd, J=7.5, 0.8 Hz, 1H), 8.17 (dd, J=7.7, 0.8 Hz, 1H),
16.03 (br. s., 1H).
Step p
4-(7-Carbamoyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-1-carboxylic
Acid tert-butyl Ester (XX) [n=0; R1=piperidin-4-yl;
X=tert-butoxycarbonyl]
Method A: to a solution of
2-(1-tert-butoxycarbonyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-
-carboxylic acid (XVIII) (3.7 g, 10.3 mmol) in
N,N-dimethylformamide (60 mL) hydroxybenzotriazole ammonium salt
(3.15 g, 20.7 mmol), 1-ethyl-3-(3'-dimethylamino)carbodiimide
hydrochloric acid salt (3.34 g, 20.7 mmol) and
diisopropylethylamine (5.3 mL, 30.9 mmol) were added. The reaction
mixture was stirred at room temperature overnight. The solvent was
removed under reduced pressure and the residue was dissolved in
ethyl acetate. The solution was washed twice with saturated sodium
carbonate aqueous solution, and the organic phase was dried over
anhydrous sodium sulfate and concentrated in vacuo. The crude was
purified by flash chromatography (dichloromethane/methanol 97:3) to
afford the title compound (2.74 g, 74%) as a white solid.
Method B: a solution of
2-(1-tert-butoxycarbonyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carbo-
xylic acid (XVIII) (5.5 g, 15.3 mmol) and carbonyldiimidazole (3.7
g, 22.8 mmol) in dry tetrahydrofuran (80 mL) was stirred at room
temperature for 4 h. Then concentrated aqueous ammonia (25 mL) was
added and the reaction mixture was left at room temperature until
the disappearance of the starting material (3 h). The solvent was
evaporated under reduced pressure and the resulting crude primary
amide (1.1 g, 20%) was employed without any further
purification.
HRMS (ESI+): calcd. for C.sub.19H.sub.26N.sub.3O.sub.4 [M+H].sup.+
360.1918. found 360.1921.
Operating according to method A, but employing suitably substituted
starting material the following compounds were obtained:
2-benzyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic Acid Amide
(I), cpd 1
[R=H; n=1; R1=phenyl; m=0; R2=null]
##STR00025##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.72-1.88 (m,
4H), 2.04-2.13 (m, 2H), 2.88-2.96 (m, 2H), 3.51 (s, 2H), 4.00-4.11
(m, 1H), 4.56 (s, 2H), 7.20-7.30 (m, 1H), 7.31-7.37 (m, 4H), 7.66
(br. s., 1H), 7.71 (dd, J=7.6, 7.4 Hz, 1H), 7.76 (dd, J=7.6, 1.5
Hz, 1H), 8.20 (dd, J=7.4, 1.5 Hz, 1H), 10.72 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.15N.sub.2O.sub.2 [M+H].sup.+
267.1128. found 267.1120.
3-oxo-2-phenethyl-2,3-dihydro-1H-isoindole-4-carboxylic Acid Amide
(I), cpd 2
[R=H; n=2; R1=phenyl; m=0; R2=null]
##STR00026##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 2.97 (t, J=7.6
Hz, 2H), 3.82 (t, J=7.6 Hz, 2H), 4.49 (s, 2H), 7.17-7.24 (m, 1H),
7.24-7.32 (m, 4H), 7.66 (br. s., 1H), 7.70 (dd, J=7.5, 7.3 Hz, 1H),
7.74 (dd, J=7.5, 1.5 Hz, 1H), 8.19 (dd, J=7.3, 1.5 Hz, 1H), 10.68
(br. s., 1H).
HRMS (ESI+): calcd. for C.sub.17H.sub.15N.sub.2O.sub.2 [M+H].sup.+
281.1285. found 281.1295.
2-[2-(3,4-dihydro-1H-isoquinolin-2-yl)-ethyl]-3-oxo-2,3-dihydro-1H-isoindo-
le-4-carboxylic Acid Amide (I), cpd 3
[R=H; n=2; R1=3,4-dihydro-1H-isoquinolin-2-yl; m=0; R2=null]
##STR00027##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 2.73-2.84 (m,
6H), 3.65 (s, 2H), 3.81 (t, J=6.2 Hz, 2H), 4.65 (s, 2H), 7.00-7.12
(m, 4H), 7.66 (br. s., 1H), 7.69 (dd, J=7.6, 7.7 Hz, 1H), 7.76 (dd,
J=7.6, 1.2 Hz, 1H), 8.19 (dd, J=7.7, 1.2 Hz, 1H), 10.75 (br. s.,
1H).
HRMS (ESI+): calcd. for C.sub.20H.sub.23N.sub.3O.sub.2 [M+H].sup.+
336.1707. found 336.1722.
3-oxo-2-(2-piperidin-1-yl-ethyl)-2,3-dihydro-1H-isoindole-4-carboxylic
Acid Amide (I), cpd 4
[R=H; n=2; R1=piperidin-1-yl; m=0; R2=null]
##STR00028##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.33-1.40 (m,
2H), 1.43-1.50 (m, 4H), 2.35-2.43 (m, 4H), 2.54 (t, J=6.3 Hz, 2H),
3.68 (t, J=6.3 Hz, 2H), 4.63 (s, 2H), 7.66 (br. s., 1H), 7.72 (dd,
J=7.7, 7.4 Hz, 1H), 7.78 (dd, J=7.4, 1.2 Hz, 1H), 8.20 (dd, J=7.7,
1.2 Hz, 1H), 10.75 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.22N.sub.3O.sub.2 [M+H].sup.+
288.1707. found 288.1712.
2-(2-morpholin-4-yl-ethyl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic
Acid Amide (I), cpd 5
[R=H; n=2; R1=morpholin-4-yl; m=0; R2=null]
##STR00029##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 2.41-2.46 (m,
4H), 2.59 (t, J=6.3 Hz, 2H), 3.52-3.57 (m, 4H), 3.71 (t, J=6.3 Hz,
2H), 4.64 (s, 2H), 7.66 (br. s., 1H), 7.72 (dd, J=7.7, 7.6 Hz, 1H),
7.78 (dd, J=7.6, 1.3 Hz, 1H), 8.20 (dd, J=7.7, 1.3 Hz, 1H), 10.73
(br. s., 1H).
HRMS (ESI+): calcd. for C.sub.15H.sub.20N.sub.3O.sub.3 [M+H].sup.+
290.1499. found 290.1507.
2-(3-morpholin-4-yl-propyl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic
Acid Amide (I), cpd 6
[R=H; n=3; R1=morpholin-4-yl; m=0; R2=null]
##STR00030##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.80 (quintet,
J=7.1 Hz, 2H), 2.28-2.38 (m, 6H), 3.47-3.54 (m, 4H), 3.61 (t, J=7.1
Hz, 2H), 4.58 (s, 2H), 7.65 (br. s., 1H), 7.71 (dd, J=7.6, 7.4 Hz,
1H), 7.77 (dd, J=7.4, 1.2 Hz, 1H), 8.20 (dd, J=7.6, 1.2 Hz, 1H),
10.76 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.22N.sub.3O.sub.3 [M+H].sup.+
304.1656. found 304.1664.
2-[2-(3,4-dihydro-2H-quinolin-1-yl)-ethyl]-3-oxo-2,3-dihydro-1H-isoindole--
4-carboxylic Acid Amide (I), cpd 7
[R=H; n=2; R1=3,4-dihydro-2H-quinolin-1-yl; m=0; R2=null]
##STR00031##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.79-1.89 (m,
2H), 2.64-2.70 (m, 2H), 3.27-3.31 (m, 2H), 3.56 (t, J=7.1 Hz, 2H),
3.76 (t, J=7.1 Hz, 2H), 4.65 (s, 2H), 6.44-6.49 (m, 1H), 6.70-6.75
(m, 1H), 6.85-6.89 (m, 1H), 6.92-6.97 (m, 1H), 7.69 (br. s., 1H),
7.72 (dd, J=7.6, 7.6 Hz, 1H), 7.77 (dd, J=7.6, 1.3 Hz 1H), 8.20
(dd, J=7.6, 1.3 Hz, 1H), 10.68 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.20H.sub.22N.sub.3O.sub.2 [M+H].sup.+
336.1707. found 336.1692.
3-oxo-2-(3-phenyl-propyl)-2,3-dihydro-1H-isoindole-4-carboxylic
Acid Amide (I), cpd 17
[R=H; n=3; R1=phenyl; m=0; R2=null]
##STR00032##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.97 (quintet,
J=7.9 Hz, 2H), 2.64 (t, J=7.9 Hz 2H), 3.61 (t, J=7.9 Hz, 2H), 4.59
(s, 2H), 7.15-7.20 (m, 1H), 7.23-7.31 (m, 4H), 7.67 (br. s., 1H),
7.72 (dd, J=7.6, 7.4 Hz, 1H), 7.77 (dd, J=7.6, 1.5 Hz, 1H), 8.21
(dd, J=7.4, 1.5 Hz, 1H), 10.74 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.18H.sub.19N.sub.2O.sub.2 [M+H].sup.+
295.1441. found 295.1433.
3-oxo-2-(2-pyridin-2-yl-ethyl)-2,3-dihydro-1H-isoindole-4-carboxylic
Acid Amide (I), cpd 18
[R=H; n=2; R1=pyrid-2-yl; m=0; R2=null]
##STR00033##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 3.12 (t, J=7.3
Hz, 2H), 3.96 (t, J=7.3 Hz, 2H), 4.52 (s, 2H), 7.23 (ddd, J=7.5,
4.9, 1.2 Hz, 1H), 7.32 (ddd, J=7.8, 1.2, 0.8 Hz, 1H), 7.65 (br. s.,
1H), 7.71 (m, 1H), 7.70 (dd, J=7.6, 7.4 Hz, 1H), 7.75 (dd, J=7.6,
1.3 Hz, 1H), 8.19 (dd, J=7.4, 1.3 Hz, 1H), 8.48 (ddd, J=4.9, 1.8,
0.8 Hz, 1H), 10.66 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.16N.sub.3O.sub.2 [M+H].sup.+
282.1237. found 282.1243.
2-[3-(3,4-dihydro-1H-isoquinolin-2-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoind-
ole-4-carboxylic Acid Amide (I), cpd 19
[R=H; n=3; R1=3,4-dihydro-1H-isoquinolin-2-yl; m=0; R2=null]
##STR00034##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.92 (quintet,
J=7.3 Hz, 2H), 2.52 (t, J=7.3 Hz, 2H), 2.64-2.70 (m, 2H), 2.76-2.82
(m, 2H), 3.57 (s, 2H), 3.64 (t, J=7.3 Hz, 2H), 4.60 (s, 2H),
7.00-7.70 (m, 4H), 7.66 (br. s., 1H), 7.70 (dd, J=7.6, 7.4 Hz, 1H),
7.75 (dd, J=7.4, 1.3 Hz, 1H), 8.19 (dd, J=7.6, 1.3 Hz, 1H), 10.76
(br. s., 1H).
HRMS (ESI+): calcd. for C.sub.2+124N.sub.3O.sub.2 [M+H].sup.+
350.1863. found 350.1866.
2-[3-(3,4-dihydro-2H-quinolin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole-
-4-carboxylic Acid Amide (I), cpd 20
[R=H; n=3; R1=3,4-dihydro-2H-quinolin-1-yl; m=0; R2=null]
##STR00035##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.80-1.87 (m,
2H), 1.90 (quintet, J=7.2 Hz, 2H), 2.65 (t, J=7.2 Hz, 2H),
3.22-3.26 (m, 2H), 3.27-3.30 (m overlapped by water signal, 2H),
3.64 (t, J=7.2 Hz, 2H), 4.59 (s, 2H), 6.42-6.47 (m, 1H), 6.54-6.58
(m, 1H), 6.83-6.87 (m, 1H), 6.89-6.94 (m, 1H), 7.67 (br. s., 1H),
7.72 (dd, J=7.6, 7.6 Hz, 1H), 7.77 (dd, J=7.6, 1.3 Hz, 1H), 8.20
(dd, J=7.6, 1.3 Hz, 1H), 10.72 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.211-124N.sub.3O.sub.2 [M+H].sup.+
350.1863. found 350.1868.
2-[3-(4-methyl-piperazin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole-4-ca-
rboxylic Acid Amide (I), cpd 21
[R=H; n=3; R1=4-methyl-piperazin-1-yl; m=0; R2=null]
##STR00036##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.80 (quintet,
J=7.2 Hz, 2H), 2.11 (s, 3H), 2.15-2.43 (br. s., 8H), 2.33 (t, J=7.2
Hz, 2H), 3.60 (t, J=7.2 Hz, 2H), 4.58 (s, 2H), 7.66 (br. s., 1H),
7.72 (dd, J=7.6, 7.4 Hz, 1H), 7.77 (dd, J=7.6, 1.3 Hz, 1H), 8.21
(dd, J=7.4, 1.3 Hz, 1H), 10.79 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.17H.sub.26N.sub.4O.sub.2 [M+H].sup.+
317.1972. found 317.1975.
3-oxo-2-[3-(4-phenyl-piperazin-1-yl)-propyl]-2,3-dihydro-1H-isoindole-4-ca-
rboxylic Acid Amide (I), cpd 22
[R=H; n=3; R1=piperazin-1-yl; m=0; R2=4-phenyl]
##STR00037##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.85 (quintet,
J=7.1 Hz, 2H), 2.39 (t, J=7.1 Hz, 2H), 3.02-3.10 (m, 4H), 3.63 (t,
J=7.1 Hz, 2H), 4.60 (s, 2H), 6.75 (t, J=7.3 Hz, 1H), 6.89 (d, J=7.9
Hz, 2H), 7.19 (dd, J=7.9, 7.3 Hz, 2H), 7.66 (br. s., 1H), 7.70 (dd,
J=7.7, 7.4 Hz, 1H), 7.76 (dd, J=7.4, 1.2 Hz, 1H), 8.20 (dd, J=7.7,
1.2 Hz, 1H), 10.78 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.22H.sub.27N.sub.4O.sub.2 [M+H].sup.+
379.2129. found 379.2145.
3-oxo-2-(3-piperidin-1-yl-propyl)-2,3-dihydro-1H-isoindole-4-carboxylic
Acid Amide Hydrochloride (I), cpd 25
[R=H; n=3; R1=piperidin-1-yl; m=0; R2=null]
##STR00038##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.29 (m, 1H),
1.50-1.73 (m, 3H), 1.75-1.85 (m, 2H), 2.00-2.09 (m, 2H), 2.79-2.92
(m, 2H), 3.03-3.14 (m, 2H), 3.40-3.50 (m, 2H), 3.66 (t, J=6.6 Hz,
2H), 4.59 (s, 2H), 7.71 (br. s., 1H), 7.74 (dd, J=7.6, 7.4 Hz, 1H),
7.80 (dd, J=7.4, 1.1 Hz, 1H), 8.21 (dd, J=7.6, 1.1 Hz, 1H), 8.93
(br. s., 1H), 10.58 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.17H.sub.24N.sub.3O.sub.2 [M+H].sup.+
302.1863. found 302.1865.
2-(3-[1,4']bipiperidinyl-1-yl-propyl)-3-oxo-2,3-dihydro-1H-isoindole-4-car-
boxylic Acid Amide Dihydrochloride (I), cpd 26
[R=H; n=3; R1=piperidin-1-yl; m=0; R2=4-piperidin-1-yl]
##STR00039##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.33-1.49 (m,
1H), 1.57-1.74 (m, 3H), 1.74-1.94 (m, 4H), 1.95-2.13 (m, 2H),
2.16-2.30 (m, 2H), 2.87-3.72 (m, 11H), 3.66 (t, J=6.5 Hz, 2H), 4.59
(s, 2H), 7.72 (br. s., 1H), 7.75 (dd, J=7.6, 6.7 Hz, 1H), 7.80 (d,
J=6.7 Hz, 1H), 8.21 (dd, J=7.6, 1.2 Hz, 1H), 9.38 (br. s., 2H),
10.58 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.22H.sub.33N.sub.4O.sub.2 [M+H].sup.+
385.2598. found 385.2611.
2-[3-(2,6-dimethyl-piperidin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole--
4-carboxylic Acid Amide Hydrochloride (I), cpd 27
[R=H; n=3; R1=2,6-dimethyl-piperidin-1-yl; m=0; R2=null]
##STR00040##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.25 (d, J=6.3
Hz, 6H), 1.40-1.55 (m, 3H), 1.80-1.91 (m, 2H), 1.95-2.05 (m, 2H),
3.02-3.46 (m, 4H), 3.68 (m, 2H), 4.63 (s, 2H), 7.70 (br. s., 1H),
7.74 (dd, J=7.6, 7.6 Hz, 1H), 7.80 (dd, J=7.6, 1.2 Hz, 1H), 8.21
(dd, J=7.6, 1.2 Hz, 1H), 8.72 (br. s., 1H), 10.58 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.28N.sub.3O.sub.2 [M+H].sup.+
330.2176. found 330.2176.
3-oxo-2-[1-(tetrahydro-pyran-4-yl)-piperidin-4-yl]-2,3-dihydro-1H-isoindol-
e-4-carboxylic Acid Amide (I), cpd 28
[R=H; n=m=0; R1=piperidin-4-yl; R2=1-(tetrahydro-pyran-4-yl)]
##STR00041##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.38-1.50 (m,
2H), 1.65-1.72 (m, 2H), 1.73-1.81 (m, 4H), 2.18-2.28 (m, 2H),
2.43-2.47 (m, 1H), 2.97-3.04 (m, 2H), 3.30 (m overlapped by water
signal, 2H), 3.89 (dd, J=11.1, 3.9 Hz, 2H), 4.02 (m, 1H), 4.55 (s,
2H), 7.66 (br. s., 1H), 7.71 (dd, J=7.6, 7.4 Hz, 1H), 7.76 (dd,
J=7.4, 1.5 Hz, 1H), 8.20 (dd, J=7.6, 1.5 Hz, 1H), 10.74 (br. s.,
1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.26N.sub.3O.sub.3 [M+H].sup.+
344.1969. found 344.1962.
2-(1-benzyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylic
Acid Amide (I), cpd 30
[R=H; n=0; R1=piperidin-4-yl; m=1; R2=phenyl]
##STR00042##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.70-1.88 (m,
4H), 1.99-2.13 (m, 2H), 2.89-2.96 (m, 2H), 3.51 (s, 2H), 4.00-4.11
(m, 1H), 4.56 (s, 2H), 7.20-7.30 (m, 1H), 7.30-7.36 (m, 4H), 7.66
(br. s., 1H), 7.71 (dd, J=7.6, 7.4 Hz, 1H), 7.76 (dd, J=7.6, 1.5
Hz, 1H), 8.20 (dd, J=7.4, 1.5 Hz, 1H), 10.72 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.21H.sub.24N.sub.3O.sub.2 [M+H].sup.+
350.1863. found 350.1874.
2-[2-(1-benzyl-piperidin-4-yl)-ethyl]-3-oxo-2,3-dihydro-1H-isoindole-4-car-
boxylic Acid Amide Hydrochloride (I), cpd 31
[R=H; n=2; R1=piperidin-4-yl; m=1; R2=phenyl]
##STR00043##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.29-1.41 (m,
2H), 1.43-1.54 (m, 1H), 1.56-1.64 (m, 2H), 1.92-2.00 (m, 2H),
2.83-2.96 (m, 2H), 3.30 (m overlapped by water signal, 2H), 3.61
(t, J=6.8 Hz, 2H), 4.25 (d, J=5.1 Hz, 2H), 4.57 (s, 2H), 7.47 (s,
5H), 7.69 (br. s., 1H), 7.72 (dd, J=7.6, 7.4 Hz, 1H), 7.77 (dd,
J=7.4, 1.3 Hz, 1H), 8.20 (dd, J=7.6, 1.3 Hz, 1H), 9.22 (br. s.,
1H), 10.68 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.23H.sub.28N.sub.3O.sub.2 [M+H].sup.+
378.2176. found 378.2178.
2-[3-(4-benzyl-piperidin-1-yl)-propyl]-3-oxo-2,3-dihydro-1H-isoindole-4-ca-
rboxylic Acid Amide (I), cpd 32
[R=H; n=3; R1=piperidin-1-yl; m=1; R2=phenyl]
##STR00044##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 0.93-1.12 (m,
2H), 1.34-1.53 (m, 3H), 1.68-1.82 (m, 4H), 2.22-2.33 (m, 2H), 2.41
(d, J=6.8 Hz, 2H), 2.75-2.85 (m, 2H), 3.59 (t, J=6.9 Hz, 2H), 4.56
(s, 2H), 7.09-7.14 (m, 2H), 7.14-7.19 (m, 1H), 7.23-7.29 (m, 2H),
7.65 (br. s., 1H), 7.72 (dd, J=7.4, 7.4 Hz, 1H), 7.77 (dd, J=7.4,
1.3 Hz, 1H), 8.21 (dd, J=7.4, 1.3 Hz, 1H), 10.78 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.24H.sub.30N.sub.3O.sub.2 [M+H].sup.+
392.2333. found 392.2346.
Step i'
3-Oxo-2-piperidin-4-yl-2,3-dihydro-1H-isoindole-4-carboxylic Acid
Amide Hydrochloride (XXI)
[n=0; R1=piperidin-4-yl]
A solution of
4-(7-carbamoyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-1-carboxylic
acid tert-butyl ester (XX) (2.7 g, 7.5 mmol) in 4M hydrochloric
acid in dioxane (18 mL, 75 mmol) was stirred at 50.degree. C. for 2
h until HPLC analysis revealed the disappearance of the starting
material. The solvent was removed under reduced pressure and the
product was dissolved in diethyl ether and filtered to obtain the
title compound (2.09 g, 95%) as its hydrochloride.
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.93-2.09 (m,
4H), 3.03-3.17 (m, 2H), 3.35-3.48 (m overlapped by water signal,
2H), 4.32-4.45 (m, 1H), 4.56 (s, 2H), 7.71 (br. s., 1H), 7.75 (dd,
J=7.5, 7.5 Hz, 1H), 7.82 (dd, J=7.5, 1.1 Hz 1H), 8.21 (dd, J=7.5,
1.1 Hz, 1H), 8.59 (br.s., 1H), 8.82 (br. s., 1H), 10.58 (br. s.,
1H).
HRMS (ESI+): calcd. for C.sub.14H.sub.18N.sub.3O.sub.2 [M+H].sup.+
260.1394. found 260.1398.
Step l'
2-(1-cyclohexyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxyli-
c Acid Amide (I), cpd 11
[R=H; n=m=0; R1=piperidin-4-yl; R2=1-cyclohexyl]
##STR00045##
Method A: to a suspension of
3-oxo-2-piperidin-4-yl-2,3-dihydro-1H-isoindole-4-carboxylic acid
amide hydrochloride (56 mg, 0.19 mmol) in dichloromethane (2 mL),
cyclohexanone (XIV) (27.5 mg, 0.28 mmol), sodium acetate (32 mg,
0.38 mmol) and methanol (0.3 mL) were added. The resultant solution
was stirred at room temperature for 5 h. Then sodium
cyanoborohydride (13 mg, 0.21 mmol) was added and the mixture was
stirred overnight. Solvents were removed under reduced pressure and
the residue was dissolved in dichloromethane and washed twice with
water. The organic phase was dried over Na.sub.2SO.sub.4 and
concentrated in vacuo and the residue was purified by flash
chromatography (dichloromethane/methanol 95:5) to give 27 mg (40%)
of
2-(1-cyclohexyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-
-carboxylic acid amide.
Method B: to a solution of
2-piperidin-4-yl-2,3-dihydro-1H-isoindole-4-carboxylic acid amide
hydrochloride (4.4 g, 14.8 mmol) and cyclohexanone (2.2 g, 22.45
mmol) in N,N-dimethylformamide (100 mL), glacial acetic acid (4.5
mL) and tetramethylammonium triacetoxyborohydride (11.8 g, 44.85
mmol) were added. The resulting solution was allowed to stir
overnight at room temperature.
The solvent was then evaporated under reduced pressure and the
resultant residue was diluted with aqueous 8% ammonia solution and
extracted with ethyl acetate. The organic phase was dried with
anhydrous sodium sulfate and concentrated. The crude was purified
by flash chromatography (dichloromethane/methanol 95:5) and
subsequently dissolved in a small amount of methanol and
precipitated with diethyl ether. The precipitate was filtered and
washed with diethyl ether to give 1.77 g of the desired product as
a white solid (35%).
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.00-1.14 (m,
1H), 1.14-1.32 (m, 4H), 1.55-1.62 (m, 1H), 1.70-1.80 (m, 8H),
2.25-2.37 (m, 3H), 2.88-2.98 (m, 2H), 3.95-4.06 (m, 1H), 4.55 (s,
2H), 7.66 (br. s., 1H), 7.71 (dd, J=7.6, 7.6 Hz, 1H), 7.76 (dd,
J=7.6, 1.5 Hz, 1H), 8.20 (dd, J=7.6, 1.5 Hz, 1H), 10.74 (br. s.,
1H).
HRMS (ESI+): calcd. for C.sub.20H.sub.28N.sub.3O.sub.2 [M+H].sup.+
342.2176. found 342.2175.
Operating according to method A, but employing suitably substituted
starting material (XIV), the following compounds were obtained:
3-oxo-2-(1-pyridin-4-ylmethyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-c-
arboxylic Acid Amide (I), cpd 8
[R=H; n=0; R1=piperidin-4-yl; m=1; R2=pyrid-4-yl]
##STR00046##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.75-1.92 (m,
4H), 2.11-2.22 (m, 2H), 2.88-2.95 (m, 2H), 3.56 (s, 2H), 4.02-4.14
(m, 1H), 4.58 (s, 2H), 7.33-7.38 (m, 2H), 7.68 (br. s., 1H), 7.73
(dd, J=7.6, 7.6 Hz, 1H), 7.78 (dd, J=7.6, 1.5 1H), 8.21 (dd, J=7.6,
1.4 Hz, 1H), 8.51-8.55 (m, 2H), 10.72 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.20H.sub.23N.sub.4O.sub.2 [M+H].sup.+
351.1816. found 351.1817.
3-oxo-2-(1-thiophen-2-ylmethyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4--
carboxylic Acid Amide (I), cpd 9
[R=H; n=0; R1=piperidin-4-yl; m=1; R2=thiophen-2-yl]
##STR00047##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.73-1.88 (m,
4H), 2.06-2.20 (m, 2H), 2.94-3.03 (m, 2H), 3.73 (s, 2H), 4.01-4.11
(m, 1H), 4.58 (s, 2H), 6.96-7.00 (m, 2H), 7.42-7.46 (m, 1H), 7.67
(br. s., 1H), 7.72 (dd, J=7.6, 7.4 Hz, 1H), 7.77 (dd, J=7.6, 1.5
Hz, 1H), 8.21 (dd, J=7.4, 1.5 Hz, 1H), 10.72 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.19H.sub.22N.sub.3O.sub.2S [M+H].sup.+
356.1427. found 356.1430.
3-oxo-2-(1-pyridin-3-ylmethyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-c-
arboxylic Acid Amide (I), cpd 10
[R=H; n=0; R1=piperidin-4-yl; m=1; R2=pyrid-3-yl]
##STR00048##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.70-1.80 (m,
4H), 2.08-2.18 (m, 2H), 2.87-2.96 (m, 2H), 3.55 (s, 2H), 4.01-4.12
(m, 1H), 4.56 (s, 2H), 7.37 (dd, J=7.7, 4.8 Hz 1H), 7.66 (br. s.,
1H), 7.71 (t, J=7.7, 7.4 Hz, 1H), 7.73 (signal overlapped by
others, 1H), 7.76 (dd, J=7.7, 1.3 Hz, 1H), 8.20 (dd, J=7.4, 1.3 Hz,
1H), 8.48 (d, J=4.8 Hz, 1H), 8.52 (s, 1H), 10.71 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.20H.sub.23N.sub.4O.sub.2 [M+H].sup.+
351.1816. found 351.1822.
2-(1-furan-2-ylmethyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-car-
boxylic Acid Amide (I), cpd 12
[R=H; n=0; R1=piperidin-4-yl; m=1; R2=fur-2-yl]
##STR00049##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.70-1.87 (m,
4H), 2.07-2.16 (m, 2H), 2.90-2.97 (m, 1H), 3.53 (s, 2H), 3.98-4.06
(m, 1H), 4.55 (s, 2H), 6.30 (d, J=2.4 Hz, 1H), 6.41 (dd, J=2.4, 1.8
Hz, 1H), 7.59 (br. s., 1H), 7.66 (br. s., 1H), 7.71 (dd, J=7.6, 7.4
Hz, 1H), 7.76 (dd, J=7.4, 1.3 Hz, 1H), 8.19 (dd, J=7.6, 1.3 Hz,
1H), 10.71 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.22N.sub.3O.sub.3 [M+H].sup.+
340.1656. found 340.1651.
3-oxo-2-(1-thiophen-3-ylmethyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4--
carboxylic Acid Amide (I), cpd 13
[R=H; n=0; R1=piperidin-4-yl; m=1; R2=thiophen-3-yl]
##STR00050##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.72-1.90 (m,
4H), 2.00-2.12 (m, 2H), 2.90-2.99 (m, 2H), 3.53 (s, 2H), 4.00-4.09
(m, 1H), 4.57 (s, 2H) 7.08 (d, J=4.6 Hz, 1H), 7.33 (br. s., 1H),
7.49 (dd, J=4.6, 2.8 Hz, 1H), 7.67 (br. s., 1H), 7.72 (dd, J=7.6,
7.4 Hz, 1H), 7.75 (dd, J=7.6, 1.3, 1H), 8.21 (dd, J=7.4, 1.3 Hz,
1H), 10.73 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.22N.sub.3O.sub.2S [M+H].sup.+
356.1427. found 356.1432.
2-(1-furan-3-ylmethyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-car-
boxylic Acid Amide (I), cpd 14
[R=H; n=0; R1=piperidin-4-yl; m=1; R2=fur-3-yl]
##STR00051##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.72-1.89 (m,
4H), 2.02-2.12 (m, 2H), 2.90-3.02 (m, 2H), 3.37 (s, 2H), 4.00-4.10
(m, 1H), 4.56 (s, 2H), 6.45 (s, 1H), 7.58 (s, 1H), 7.62 (s, 1H),
7.67 (br. s., 1H), 7.72 (dd, J=7.6, 7.4 Hz, 1H), 7.76 (dd, J=7.4,
1.5 Hz, 1H), 8.21 (dd, J=7.6, 1.5 Hz, 1H), 10.73 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.22N.sub.3O.sub.3 [M+H].sup.+
340.1656. found 340.1649.
3-oxo-2-(1-pyridin-2-ylmethyl-piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-c-
arboxylic Acid Amide (I), cpd 15
[R=H; n=0; R1=piperidin-4-yl; m=1; R2=pyrid-2-yl]
##STR00052##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.74-1.93 (m,
4H), 2.12-2.28 (m, 2H), 2.91-3.00 (m, 2H), 3.65 (s, 2H), 4.02-4.13
(m, 1H), 4.58 (s, 2H), 7.28 (dd, J=6.8, 4.8 Hz, 1H), 7.47 (d, J=7.8
Hz, 1H), 7.67 (br. s., 1H), 7.73 (dd, J=7.4, 7.4 Hz, 1H), 7.76-7.83
(m, 2H), 8.21 (dd, J=7.4, 1.3 Hz, 1H), 8.51 (d, J=4.8 Hz, 1H),
10.73 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.20H.sub.23N.sub.4O.sub.2 [M+H].sup.+
351.1816. found 351.1815.
3-oxo-2-[1-(1H-pyrrol-2-ylmethyl)-piperidin-4-yl]-2,3-dihydro-1H-isoindole-
-4-carboxylic Acid Amide (I), cpd 16
[R=H; n=0; R1=piperidin-4-yl; m=1; R2=1H-pyrrol-2-yl]
##STR00053##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 1.70-1.88 (m,
4H), 1.96-2.09 (m, 2H), 2.88-2.99 (m, 2H), 3.44 (s, 2H), 3.94-4.09
(m, 1H), 4.55 (s, 2H), 5.89 (br. s., 1H), 5.94 (br. s., 1H), 6.65
(br. s., 1H), 7.67 (br. s., 1H), 7.72 (dd, J=7.6, 7.4 Hz, 1H), 7.77
(dd, J=7.6, 1.3 Hz, 1H), 8.20 (dd, J=7.4, 1.3 Hz, 1H), 10.65 (br.
s., 1H), 10.73 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.16H.sub.23N.sub.4O.sub.2 [M+H].sup.+
339.1816. found 339.1812.
2-(1-cyclopropylmethyl-piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-ca-
rboxylic Acid Amide (I), cpd 24
[R=H; n=0; R1=piperidin-4-yl; m=1; R2=cyclopropyl]
##STR00054##
.sup.1H NMR (400.5 MHz, DMSO-d.sub.6) .delta. ppm 0.06-0.12 (m,
2H), 0.44-0.50 (m, 2H), 0.80-0.89 (m, 1H), 1.72-1.88 (m, 4H),
2.00-2.11 (m, 2H), 2.21 (d, J=6.3 Hz, 2H), 3.04-3.13 (m, 2H),
3.98-4.09 (m, 1H), 4.56 (s, 2H), 7.66 (br. s., 1H), 7.72 (dd,
J=7.6, 7.6 Hz, 1H), 7.77 (dd, J=7.6, 1.2 Hz, 1H), 8.20 (dd, J=7.6,
1.2 Hz, 1H), 10.73 (br. s., 1H).
HRMS (ESI+): calcd. for C.sub.18H.sub.24N.sub.3O.sub.2 [M+H].sup.+
314.1863. found 314.1860.
* * * * *