U.S. patent application number 15/425119 was filed with the patent office on 2017-05-25 for phosphate compounds, a process for their preparation and pharmaceutical compositions containing them.
The applicant listed for this patent is LES LABORATOIRES SERVIER, VERNALIS (R&D) LTD. Invention is credited to I-Jen Chen, James Edward Paul Davidson, Guillaume De Nanteuil, Olivier Geneste, Anne-Francoise Guillouzic, Jean-Michel Henlin, Thierry Le Diguarher, Arnaud LE TIRAN, James Brooke Murray, Jerome-Benoit Starck.
Application Number | 20170143746 15/425119 |
Document ID | / |
Family ID | 50288170 |
Filed Date | 2017-05-25 |
United States Patent
Application |
20170143746 |
Kind Code |
A1 |
LE TIRAN; Arnaud ; et
al. |
May 25, 2017 |
PHOSPHATE COMPOUNDS, A PROCESS FOR THEIR PREPARATION AND
PHARMACEUTICAL COMPOSITIONS CONTAINING THEM
Abstract
Compounds of formula (I): ##STR00001## wherein X, Y, A.sub.1,
A.sub.2, R.sub.a, R.sub.b, R.sub.c, R.sub.d, R.sub.3, R.sub.4, T
and R.sub.5 are as defined in the description. Medicinal products
containing the same which are useful in treating pathologies
involving a deficit in apoptosis, such as cancer, auto-immune
diseases, and diseases of the immune system.
Inventors: |
LE TIRAN; Arnaud; (Croissy
sur Seine, FR) ; Le Diguarher; Thierry; (Saint Denis
de I'Hotel, FR) ; Starck; Jerome-Benoit;
(Rueil-Malmaison, FR) ; Henlin; Jean-Michel;
(Suresnes, FR) ; Guillouzic; Anne-Francoise;
(Nanterre, FR) ; De Nanteuil; Guillaume;
(Suresnes, FR) ; Geneste; Olivier;
(Rueil-Malmaison, FR) ; Davidson; James Edward Paul;
(Great Shelford, GB) ; Murray; James Brooke;
(Linton, GB) ; Chen; I-Jen; (Cambridge,
GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
LES LABORATOIRES SERVIER
VERNALIS (R&D) LTD |
Suresnes Cedex
Winnersh |
|
FR
GB |
|
|
Family ID: |
50288170 |
Appl. No.: |
15/425119 |
Filed: |
February 6, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14800871 |
Jul 16, 2015 |
9597341 |
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15425119 |
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14337577 |
Jul 22, 2014 |
9115159 |
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14800871 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07F 9/6561 20130101;
C07F 9/09 20130101; A61P 37/00 20180101; C07F 9/65583 20130101;
A61P 35/02 20180101; A61P 43/00 20180101; A61N 5/10 20130101; A61P
35/00 20180101; A61K 31/675 20130101; A61K 45/06 20130101; A61N
2005/1098 20130101; A61P 37/02 20180101 |
International
Class: |
A61K 31/675 20060101
A61K031/675 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 23, 2013 |
FR |
1357259 |
Claims
1. A method of treating immune and auto-immune diseases in a
subject in need thereof, comprising administration of an effective
amount of a phosphate compound of formula (I): ##STR00011##
wherein: X and Y represent a carbon atom or a nitrogen atom,
wherein X and Y may not simultaneously represent two carbons atoms
or two nitrogen atoms; A.sub.1 and A.sub.2, together with the atoms
carrying them, form an optionally substituted, aromatic or
non-aromatic heterocycle Het composed of 5, 6 or 7 ring members
which may have, in addition to the nitrogen represented by X or by
Y, from one to 3 heteroatoms selected independently from oxygen,
sulphur and nitrogen, wherein the nitrogen atom may be substituted
by a hydrogen atom, a linear or branched (C.sub.1-C.sub.6)alkyl
group or a --C(O)--O-Alk group wherein Alk is a linear or branched
(C.sub.1-C.sub.6)alkyl group, or A.sub.1 and A.sub.2 independently
of one another represent a hydrogen atom, a linear or branched
(C.sub.1-C.sub.6)polyhaloalkyl, a linear or branched
(C.sub.1-C.sub.6)alkyl group or a cycloalkyl; T represents a
hydrogen atom, a linear or branched (C.sub.1-C.sub.6)alkyl group
optionally substituted by from one to three halogen atoms, a
(C.sub.1-C.sub.4)alkyl-NR.sub.1R.sub.2 group, or a
(C.sub.1-C.sub.4)alkyl-OR.sub.6 group; R.sub.1 and R.sub.2
independently of one another represent a hydrogen atom or a linear
or branched (C.sub.1-C.sub.6)alkyl group, or R.sub.1 and R.sub.2
together with the nitrogen atom carrying them form a
heterocycloalkyl; R.sub.3 represents a linear or branched
(C.sub.1-C.sub.6)alkyl group, a linear or branched
(C.sub.2-C.sub.6)alkenyl group, a linear or branched
(C.sub.2-C.sub.6)alkynyl group, a cycloalkyl group, a
(C.sub.3-C.sub.10)cycloalkyl-(C.sub.1-C.sub.6)alkyl group wherein
the alkyl moiety is linear or branched, a heterocycloalkyl group,
an aryl group or a heteroaryl group, wherein one or more of the
carbon atoms of the preceding groups, or of their possible
substituents, may be deuterated; R.sub.4 represents a phenyl
substituted in the para position by a group of formula
--OPO(OM)(OM'), --OPO(OM)(O.sup.-M.sub.1.sup.+),
--OPO(O.sup.-M.sub.1.sup.+)(O.sup.-M.sub.2.sup.+),
--OPO(O.sup.-)(O.sup.-)M.sub.3.sup.2+,
--OPO(OM)(O[CH.sub.2CH.sub.2O].sub.nCH.sub.3), or
--OPO(O.sup.-M.sub.1.sup.+)(O[CH.sub.2CH.sub.2O].sub.nCH.sub.3), or
R.sub.4 represents a pyrimidin-5-yl group substituted in the para
position by a group of formula
--OPO(O.sup.-M.sub.1.sup.+)(O.sup.-M.sub.2.sup.+), wherein M and M'
independently of one another represent a hydrogen atom, a linear or
branched (C.sub.1-C.sub.6)alkyl group, a linear or branched
(C.sub.2-C.sub.6)alkenyl group, a linear or branched
(C.sub.2-C.sub.6)alkynyl group, a cycloalkyl or a heterocycloalkyl
both composed of 5 or 6 ring members, while M.sub.1.sup.+ and
M.sub.2.sup.+ independently of one another represent a
pharmaceutically acceptable monovalent cation, and M.sub.3.sup.2+
represents a pharmaceutically acceptable divalent cation and n is
an integer from 1 to 5, wherein the phenyl group may optionally be
substituted by one or more halogen atoms; R.sub.5 represents a
hydrogen atom, a halogen atom, a linear or branched
(C.sub.1-C.sub.6)alkyl group, or a linear or branched
(C.sub.1-C.sub.6)alkoxy group; R.sub.6 represents a hydrogen atom
or a linear or branched (C.sub.1-C.sub.6)alkyl group; R.sub.a,
R.sub.b, R.sub.c and R.sub.d, each independently of the others,
represent, a hydrogen atom, a linear or branched
(C.sub.1-C.sub.6)alkyl, a linear or branched
(C.sub.2-C.sub.6)alkenyl, a linear or branched
(C.sub.2-C.sub.6)alkynyl, an aryl, a heteroaryl, a halogen atom, a
linear or branched (C.sub.1-C.sub.6)alkoxy group, a hydroxy group,
a linear or branched (C.sub.1-C.sub.6)polyhalo alkyl group, a
trifluoromethoxy group, --NR.sub.7R.sub.7', nitro,
R.sub.7--CO--(C.sub.0-C.sub.6)alkyl-,
R.sub.7--CO--NH--(C.sub.0-C.sub.6)alkyl-,
NR.sub.7R.sub.7'--CO--(C.sub.0-C.sub.6)alkyl-,
NR.sub.7R.sub.7'--CO--(C.sub.0-C.sub.6)alkyl-O--,
R.sub.7--SO.sub.2--NH--(C.sub.0-C.sub.6)alkyl-,
R.sub.7--NH--CO--NH--(C.sub.0-C.sub.6)alkyl-,
R.sub.7--O--CO--NH--(C.sub.0-C.sub.6)alkyl-, a heterocycloalkyl
group, or the substituents of one of the pairs (R.sub.a,R.sub.b),
(R.sub.b,R.sub.c) or (R.sub.c,R.sub.d) together with the carbon
atoms carrying them form a ring composed of from 5 to 7 ring
members, which ring may have from one to 2 hetero atoms selected
from oxygen and sulphur, wherein one or more carbon atoms of the
ring defined hereinbefore may be deuterated or substituted by from
one to 3 groups selected from halogen and linear or branched
(C.sub.1-C.sub.6)alkyl; R.sub.7 and R.sub.7' independently of one
another represent a hydrogen atom, a linear or branched
(C.sub.1-C.sub.6)alkyl, a linear or branched
(C.sub.2-C.sub.6)alkenyl, a linear or branched
(C.sub.2-C.sub.6)alkynyl, an aryl or a heteroaryl, or R.sub.7 and
R.sub.7' together with nitrogen atom carrying them form a
heterocycle composed of from 5 to 7 ring members, it being
understood that: "aryl" means a phenyl, naphthyl, biphenyl or
indenyl group, "heteroaryl" means any mono- or bi-cyclic group
composed of from 5 to 10 ring members, having at least one aromatic
moiety and containing from 1 to 4 hetero atoms selected from
oxygen, sulphur and nitrogen (including quaternary nitrogens),
"cycloalkyl" means any mono- or bi-cyclic, non-aromatic,
carbocyclic group containing from 3 to 10 ring members,
"heterocycloalkyl" means any mono- or bi-cyclic, non-aromatic,
condensed or spiro group composed of 3 to 10 ring members and
containing from 1 to 3 hetero atoms selected from oxygen, sulphur,
SO, SO.sub.2 and nitrogen, wherein the aryl, heteroaryl, cycloalkyl
and heterocycloalkyl groups so defined and the groups alkyl,
alkenyl, alkynyl and alkoxy may be optionally substituted by from 1
to 3 groups selected from optionally substituted, linear or
branched (C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)spiro, linear or
branched, optionally substituted (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)alkyl-S--, hydroxy, oxo (or N-oxide where
appropriate), nitro, cyano, --COOR', --OCOR', NR'R'', linear or
branched (C.sub.1-C.sub.6)polyhaloalkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkylsulphonyl, halogen, optionally substituted
aryl, heteroaryl, aryloxy, arylthio, cycloalkyl, heterocycloalkyl
optionally substituted by one or more halogen atoms or alkyl
groups, wherein R' and R'' independently of one another represent a
hydrogen atom or an optionally substituted, linear or branched
(C.sub.1-C.sub.6)alkyl group, wherein the Het group defined in
formula (I) may be optionally substituted by from one to three
groups selected from linear or branched (C.sub.1-C.sub.6)alkyl,
hydroxy, linear or branched (C.sub.1-C.sub.6)alkoxy,
NR.sub.1'R.sub.1'' and halogen, wherein R.sub.1' and R.sub.1''
independently of one another represent a hydrogen atom or an
optionally substituted, linear or branched (C.sub.1-C.sub.6)alkyl
group, or an enantiomer, a diastereoisomer, or an addition salt
thereof with a pharmaceutically acceptable acid or base.
2. The method according to claim 1, wherein R.sub.4 represents
phenyl substituted in the para position by a group of formula
--OPO(OM)(OM'), --OPO(OM)(O.sup.-M.sub.1.sup.+),
--OPO(O.sup.-M.sub.1.sup.+)(O.sup.-M.sub.2.sup.+),
--OPO(O.sup.-)(O.sup.-)M.sub.3.sup.2+,
--OPO(OM)(O[CH.sub.2CH.sub.2O].sub.nCH.sub.3), or
--OPO(O.sup.-M.sub.1.sup.+)(O[CH.sub.2CH.sub.2O].sub.nCH.sub.3),
wherein M and M' independently of one another represent a hydrogen
atom, a linear or branched (C.sub.1-C.sub.6)alkyl group, a linear
or branched (C.sub.2-C.sub.6)alkenyl group, a linear or branched
(C.sub.2-C.sub.6)alkynyl group, a cycloalkyl or a heterocycloalkyl
both composed of 5 or 6 ring members, and wherein M.sub.1.sup.+ and
M.sub.2.sup.+ independently of one another represent a
pharmaceutically acceptable monovalent cation, and M.sub.3.sup.2+
represents a pharmaceutically acceptable divalent cation and n is
an integer from 1 to 5, wherein the phenyl group may optionally be
substituted by one or more halogen atoms.
3. The method according to claim 1, wherein R.sub.4 represents
phenyl substituted in the para position by a group of formula
--OPO(O.sup.-Na.sup.+)(O.sup.-Na.sup.+).
4. The method according to claim 1, wherein X represents a carbon
atom and Y represents a nitrogen atom.
5. The method according to claim 1, wherein the group: ##STR00012##
represents a 5,6,7,8-tetrahydroindolizine, an indolizine or a
dimethylated pyrrole.
6. The method according to claim 1, wherein T represents a methyl,
(morpholin-4-yl)methyl or 3-(morpholin-4-yl)propyl group.
7. The method according to claim 1, wherein R.sub.a and R.sub.d
each represent a hydrogen atom and (R.sub.b,R.sub.c), together with
the carbon atoms carrying them, form a 1,3-dioxolane group or a
1,4-dioxane group; or R.sub.a, R.sub.c and R.sub.d each represent a
hydrogen atom and R.sub.b represents a hydrogen atom or a halogen
atom.
8. The method according to claim 1, wherein R.sub.a and R.sub.d
each represent a hydrogen atom, R.sub.b represents a halogen atom
and R.sub.c represents a methoxy group.
9. The method according to claim 1, wherein R.sub.a, R.sub.b and
R.sub.d each represent a hydrogen atom and R.sub.c represents a
NR.sub.7R.sub.7'--CO--(C.sub.0-C.sub.6)alkyl-O-- group.
10. The method according to claim 1, wherein R.sub.3 represents a
group selected from phenyl, 1H-indole, 1H-pyrrolo[2,3-b]pyridine,
pyridine, 1H-pyrazole, 1H-pyrrole and
2,3-dihydro-1H-pyrrolo[2,3-b]pyridine, those groups optionally
having one or more substituents selected from linear or branched
(C.sub.1-C.sub.6)alkyl, cyano and trideuteriomethyl.
11. The method according to claim 1, wherein the compound of
formula (I) is selected from:
4-[{[3-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl-
]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl-
}(phenyl)amino]phenyl disodium phosphate,
4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-
-yl)amino]phenyl disodium phosphate,
4-({[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}[1-(trideu-
teriomethyl)-1H-pyrazol-4-yl]amino)phenyl disodium phosphate,
4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1-
,2-dimethyl-1H-pyrrol-3-yl)amino]phenyl disodium phosphate,
4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1-
-methyl-1H-pyrrol-3-yl)amino]phenyl disodium phosphate,
4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl--
1H-pyrazol-4-yl)amino]phenyl disodium phosphate,
4-[(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl){[5-(5-fluoro-2-{[(3S)-3-(morphol-
in-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimeth-
yl-1H-pyrrol-3-yl]carbonyl}amino]phenyl disodium phosphate, and
4-[{[5-(5-fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl--
1H-pyrazol-4-yl)amino]phenyl disodium phosphate, or an enantiomer,
a diastereoisomer, or an addition salt thereof with a
pharmaceutically acceptable acid or base.
12. The method according to claim 1, wherein the compound of
formula (I) is administered as a pharmaceutical composition
comprising the compound of formula (I) in combination with one or
more pharmaceutically acceptable excipients.
Description
[0001] The present invention relates to new phosphate compounds, to
a process for their preparation and to pharmaceutical compositions
containing them.
[0002] The compounds of the present invention are new and have very
valuable pharmacological and pharmacokinetic characteristics for
use in the field of apoptosis and cancerology.
[0003] Apoptosis, or programmed cell death, is a physiological
process that is crucial for embryonic development and maintenance
of tissue homeostasis.
[0004] Apoptotic-type cell death involves morphological changes
such as condensation of the nucleus, DNA fragmentation and also
biochemical phenomena such as the activation of caspases which
cause damage to key structural components of the cell, so inducing
its disassembly and death. Regulation of the process of apoptosis
is complex and involves the activation or repression of several
intracellular signaling pathways (Cory S. et al., Nature Review
Cancer, 2002, 2, 647-656).
[0005] Deregulation of apoptosis is involved in certain
pathologies. Increased apoptosis is associated with
neurodegenerative diseases such as Parkinson's disease, Alzheimer's
disease and ischaemia. Conversely, deficits in the implementation
of apoptosis play a significant role in the development of cancers
and their chemoresistance, in auto-immune diseases, inflammatory
diseases and viral infections. Accordingly, absence of apoptosis is
one of the phenotypic signatures of cancer (Hanahan D. et al., Cell
2000, 100, 57-70).
[0006] The anti-apoptotic proteins of the Bcl-2 family are
associated with numerous pathologies. The involvement of proteins
of the Bcl-2 family is described in numerous types of cancer, such
as colorectal cancer, breast cancer, small-cell lung cancer,
non-small-cell lung cancer, bladder cancer, ovarian cancer,
prostate cancer, chronic lymphoid leukaemia, follicular lymphoma,
myeloma, etc. Overexpression of the anti-apoptotic proteins of the
Bcl-2 family is involved in tumorigenesis, in resistance to
chemotherapy and in the clinical prognosis of patients affected by
cancer. There is, therefore, a therapeutic need for compounds that
inhibit the anti-apoptotic activity of the proteins of the Bcl-2
family.
[0007] In addition to being new, the compounds of the present
invention have pharmacological and pharmacokinetic properties
making it possible to use them in pathologies involving a defect in
apoptosis, such as, for example, in the treatment of cancer,
auto-immune diseases and diseases of the immune system.
[0008] The present invention relates more especially to a phosphate
compound of formula (I):
##STR00002##
[0009] wherein: [0010] X and Y represent a carbon atom or a
nitrogen atom, it being understood that they may not simultaneously
represent two carbons atoms or two nitrogen atoms, [0011] A.sub.1
and A.sub.2, together with the atoms carrying them, form an
optionally substituted, aromatic or non-aromatic heterocycle Het
composed of 5, 6 or 7 ring members which may contain, in addition
to the nitrogen represented by X or by Y, from one to 3 hetero
atoms selected independently from oxygen, sulphur and nitrogen, it
being understood that the nitrogen in question may be substituted
by a group representing a hydrogen atom, a linear or branched
(C.sub.1-C.sub.6)alkyl group or a group --C(O)--O-Alk wherein Alk
is a linear or branched (C.sub.1-C.sub.6)alkyl group, [0012] or
A.sub.1 and A.sub.2 independently of one another represent a
hydrogen atom, a linear or branched (C.sub.1-C.sub.6)polyhaloalkyl,
a linear or branched (C.sub.1-C.sub.6)alkyl group or a cycloalkyl,
[0013] T represents a hydrogen atom, a linear or branched
(C.sub.1-C.sub.6)alkyl group optionally substituted by from one to
three halogen atoms, a group
(C.sub.1-C.sub.4)alkyl-NR.sub.1R.sub.2, or a group
(C.sub.1-C.sub.4)alkyl-OR.sub.6, [0014] R.sub.1 and R.sub.2
independently of one another represent a hydrogen atom or a linear
or branched (C.sub.1-C.sub.6)alkyl group, [0015] or R.sub.1 and
R.sub.2 form with the nitrogen atom carrying them a
heterocycloalkyl, [0016] R.sub.3 represents a linear or branched
(C.sub.1-C.sub.6)alkyl group, a linear or branched
(C.sub.2-C.sub.6)alkenyl group, a linear or branched
(C.sub.2-C.sub.6)alkynyl group, a cycloalkyl group, a
(C.sub.3-C.sub.10)cycloalkyl-(C.sub.1-C.sub.6)alkyl group wherein
the alkyl moiety is linear or branched, a heterocycloalkyl group,
an aryl group or a heteroaryl group, it being understood that one
or more of the carbon atoms of the preceding groups, or of their
possible substituents, may be deuterated, [0017] R.sub.4 represents
an aryl group, a heteroaryl group, a cycloalkyl group or a linear
or branched (C.sub.1-C.sub.6)alkyl group, it being understood that
one or more of the carbon atoms of the preceding groups, or of
their possible substituents, may be deuterated, [0018] R.sub.5
represents a hydrogen or halogen atom, a linear or branched
(C.sub.1-C.sub.6)alkyl group, or a linear or branched
(C.sub.1-C.sub.6)alkoxy group, [0019] R.sub.6 represents a hydrogen
atom or a linear or branched (C.sub.1-C.sub.6)alkyl group, [0020]
R.sub.a, R.sub.b, R.sub.c and R.sub.d, each independently of the
others, represent R.sub.7, a halogen atom, a linear or branched
(C.sub.1-C.sub.6)alkoxy group, a hydroxy group, a linear or
branched (C.sub.1-C.sub.6)polyhaloalkyl group, a trifluoromethoxy
group, --NR.sub.7R.sub.7', nitro,
R.sub.7--CO--(C.sub.0-C.sub.6)alkyl-,
R.sub.7--CO--NH--(C.sub.0-C.sub.6)alkyl-,
NR.sub.7R.sub.7'--CO--(C.sub.0-C.sub.6)alkyl-,
NR.sub.7R.sub.7'--CO--(C.sub.0-C.sub.6)alkyl-O--,
R.sub.7--SO.sub.2--NH---(C.sub.0-C.sub.6)alkyl-,
R.sub.7--NH--CO--NH--(C.sub.0-C.sub.6)alkyl-,
R.sub.7--O--CO--NH--(C.sub.0-C.sub.6)alkyl-, a heterocycloalkyl
group, or the substituents of one of the pairs (R.sub.a,R.sub.b),
(R.sub.b,R.sub.c) or (R.sub.c,R.sub.d) form together with the
carbon atoms carrying them a ring composed of from 5 to 7 ring
members, which may contain from one to 2 hetero atoms selected from
oxygen and sulphur, it also being understood that one or more
carbon atoms of the ring defined hereinbefore may be deuterated or
substituted by from one to 3 groups selected from halogen and
linear or branched (C.sub.1-C.sub.6)alkyl, [0021] R.sub.7 and
R.sub.7' independently of one another represent a hydrogen, a
linear or branched (C.sub.1-C.sub.6)alkyl, a linear or branched
(C.sub.2-C.sub.6)alkenyl, a linear or branched
(C.sub.2-C.sub.6)alkynyl, an aryl or a heteroaryl, or R.sub.7 and
R.sub.7' together with nitrogen atom carrying them form a
heterocycle composed of from 5 to 7 ring members,
[0022] the compound of formula (I) being such that at least one of
the carbon atoms contained in it is substituted by one of the
following phosphate groups: --OPO(OM)(OM'),
--OPO(OM)(O.sup.-M.sub.1.sup.+),
--OPO(O.sup.-M.sub.1.sup.+)(O.sup.-M.sub.2.sup.+),
--OPO(O.sup.-)(O.sup.-)M.sub.3.sup.2+,
--OPO(OM)(O[CH.sub.2CH.sub.2O].sub.nCH.sub.3), or
--OPO(O.sup.-M.sub.1.sup.+)(O[CH.sub.2CH.sub.2O].sub.nCH.sub.3),
wherein M and M' independently of one another represent a hydrogen
atom, a linear or branched (C.sub.1-C.sub.6)alkyl group, a linear
or branched (C.sub.2-C.sub.6)alkenyl group, a linear or branched
(C.sub.2-C.sub.6)alkynyl group, a cycloalkyl or a heterocycloalkyl
both composed of 5 or 6 ring members, while M.sub.1.sup.+ and
M.sub.2.sup.+ independently of one another represent a
pharmaceutically acceptable monovalent cation, and M.sub.3.sup.2+
represents a pharmaceutically acceptable divalent cation and n is
an integer from 1 to 5,
[0023] it being understood that: [0024] "aryl" means a phenyl,
naphthyl, biphenyl or indenyl group, [0025] "heteroaryl" means any
mono- or bi-cyclic group composed of from 5 to 10 ring members,
having at least one aromatic moiety and containing from 1 to 4
hetero atoms selected from oxygen, sulphur and nitrogen (including
quaternary nitrogens), [0026] "cycloalkyl" means any mono- or
bi-cyclic, non-aromatic, carbocyclic group containing from 3 to 10
ring members, [0027] "heterocycloalkyl" means any mono- or
bi-cyclic, non-aromatic, condensed or spiro group composed of 3 to
10 ring members and containing from 1 to 3 hetero atoms selected
from oxygen, sulphur, SO, SO.sub.2 and nitrogen,
[0028] it being possible for the aryl, heteroaryl, cycloalkyl and
heterocycloalkyl groups so defined and the groups alkyl, alkenyl,
alkynyl and alkoxy to be substituted by from 1 to 3 groups selected
from optionally substituted, linear or branched
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)spiro, linear or branched,
optionally substituted (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)alkyl-S--, hydroxy, oxo (or N-oxide where
appropriate), nitro, cyano, --COOR', --OCOR', NR'R'', linear or
branched (C.sub.1-C.sub.6)polyhalo alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkylsulphonyl, halogen, optionally substituted
aryl, heteroaryl, aryloxy, arylthio, cycloalkyl, heterocycloalkyl
optionally substituted by one or more halogen atoms or alkyl
groups, it being understood that R' and R'' independently of one
another represent a hydrogen atom or an optionally substituted,
linear or branched (C.sub.1-C.sub.6)alkyl group,
[0029] it being possible for the Het group defined in formula (I)
to be substituted by from one to three groups selected from linear
or branched (C.sub.1-C.sub.6)alkyl, hydroxy, linear or branched
(C.sub.1-C.sub.6)alkoxy, NR.sub.1'R.sub.1'' and halogen, it being
understood that R.sub.1' and R.sub.1'' are as defined for the
groups R' and R'' mentioned hereinbefore,
[0030] to its enantiomers and diastereoisomers, and to addition
salts thereof with a pharmaceutically acceptable acid or base.
[0031] Among the pharmaceutically acceptable acids there may be
mentioned, without implying any limitation, hydrochloric acid,
hydrobromic acid, sulphuric acid, phosphonic acid, acetic acid,
trifluoroacetic acid, lactic acid, pyruvic acid, malonic acid,
succinic acid, glutaric acid, fumaric acid, tartaric acid, maleic
acid, citric acid, ascorbic acid, oxalic acid, methanesulphonic
acid, camphoric acid etc.
[0032] Among the pharmaceutically acceptable bases there may be
mentioned, without implying any limitation, sodium hydroxide,
potassium hydroxide, triethylamine, tert-butylamine etc.
[0033] Preferred compounds of the invention include compounds of
formula (I) wherein R.sub.4 represents phenyl substituted in the
para position by a group of formula --OPO(OM)(OM'),
--OPO(OM)(O.sup.-M.sub.1.sup.+),
--OPO(O.sup.-M.sub.1.sup.+)(O.sup.-M.sub.2.sup.+),
--OPO(O.sup.-)(O.sup.-)M.sub.3.sup.2+,
--OPO(OM)(O[CH.sub.2CH.sub.2O].sub.nCH.sub.3), or
--OPO(O.sup.-M.sub.1.sup.+)(O[CH.sub.2CH.sub.2O].sub.nCH.sub.3),
wherein M and M' independently of one another represent a hydrogen
atom, a linear or branched (C.sub.1-C.sub.6)alkyl group, a linear
or branched (C.sub.2-C.sub.6)alkenyl group, a linear or branched
(C.sub.2-C.sub.6)alkynyl group, a cycloalkyl or a heterocycloalkyl
both composed of 5 or 6 ring members, while M.sub.1.sup.+ and
M.sub.2.sup.+ independently of one another represent a
pharmaceutically acceptable monovalent cation, and M.sub.3.sup.2+
represents a pharmaceutically acceptable divalent cation and n is
an integer from 1 to 5, it being understood that the phenyl group
may optionally be substituted by one or more halogen atoms.
[0034] Preference is given to compounds of formula (I) wherein
R.sub.4 represents a phenyl or a pyrimidin-5-yl group, both
substituted in the para position by a group of formula
--OPO(O.sup.-M.sub.1.sup.+)(O.sup.-M.sub.2.sup.+), and even more
especially by a group of formula
--OPO(O.sup.-Na.sup.+)(O.sup.-Na.sup.+).
[0035] Advantageously, X represents a carbon atom and Y represents
a nitrogen atom. Even more advantageously, the group:
##STR00003##
[0036] represents a 5,6,7,8-tetrahydroindolizine, an indolizine or
a dimethylated pyrrole.
[0037] T preferably represents a methyl, (morpholin-4-yl)methyl or
3-(morpholin-4-yl)propyl group.
[0038] In preferred compounds of the invention, R.sub.a and R.sub.d
each represent a hydrogen atom and (R.sub.b,R.sub.c), together with
the carbon atoms carrying them, form a 1,3-dioxolane group or a
1,4-dioxane group; or R.sub.a, R.sub.c and R.sub.d each represent a
hydrogen atom and R.sub.b represents a hydrogen or a halogen.
[0039] In another embodiment of the invention, R.sub.a and R.sub.d
each represent a hydrogen atom, R.sub.b represents a halogen atom
and R.sub.c a methoxy group.
[0040] Alternatively, R.sub.a, R.sub.b and R.sub.d each
advantageously represent a hydrogen atom and R.sub.c represents a
group NR.sub.7R.sub.7'--CO--(C.sub.0-C.sub.6)alkyl-O--, and even
more preferably R.sub.c represents a 2-oxo-2-(piperidin-1-yl)ethoxy
group.
[0041] Furthermore, R.sub.3 advantageously represents a group
selected from phenyl, 1H-indole, 1H-pyrrolo[2,3-b]pyridine,
pyridine, 1H-pyrazole, 1H-pyrrole and
2,3-dihydro-1H-pyrrolo[2,3-b]pyridine, those groups optionally
having one or more substituents selected from linear or branched
(C.sub.1-C.sub.6)alkyl (more preferably methyl), cyano and
trideuteriomethyl.
[0042] Among the preferred compounds of the invention there may be
mentioned: [0043]
4-[{[3-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl-
]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl-
}(phenyl)amino]phenyl disodium phosphate, [0044]
4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-
-yl)amino]phenyl disodium phosphate, [0045]
4-({[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}[1-(trideu-
teriomethyl)-1H-pyrazol-4-yl]amino)phenyl disodium phosphate,
[0046]
4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1-
,2-dimethyl-1H-pyrrol-3-yl)amino]phenyl disodium phosphate, [0047]
4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1-
-methyl-1H-pyrrol-3-yl)amino]phenyl disodium phosphate, [0048]
4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl--
1H-pyrazol-4-yl)amino]phenyl disodium phosphate, [0049]
4-[(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl){[5-(5-fluoro-2-{[(3S)-3-(morphol-
in-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimeth-
yl-1H-pyrrol-3-yl]carbonyl}amino]phenyl disodium phosphate, [0050]
4-[{[5-(5-fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-
-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl--
1H-pyrazol-4-yl)amino]phenyl disodium phosphate,
[0051] their enantiomers and diastereoisomers, and addition salts
thereof with a pharmaceutically acceptable acid or base.
[0052] Pharmacokinetic study of the phosphate compounds of formula
(I) showed that they were converted in vivo into compounds of
formula (I') characterised in that the phosphate function was
metabolised into a hydroxy function. The compounds of formula (I)
accordingly behave as prodrugs of compounds of formula (I') having
the following formula:
##STR00004##
[0053] wherein: [0054] X and Y represent a carbon atom or a
nitrogen atom, it being understood that they may not simultaneously
represent two carbons atoms or two nitrogen atoms, [0055] A.sub.1
and A.sub.2, together with the atoms carrying them, form an
optionally substituted, aromatic or non-aromatic heterocycle Het
composed of 5, 6 or 7 ring members which may contain, in addition
to the nitrogen represented by X or by Y, from one to 3 hetero
atoms selected independently from oxygen, sulphur and nitrogen, it
being understood that the nitrogen in question may be substituted
by a group representing a hydrogen atom, a linear or branched
(C.sub.1-C.sub.6)alkyl group or a group --C(O)--O-Alk wherein Alk
is a linear or branched (C.sub.1-C.sub.6)alkyl group, [0056] or
A.sub.1 and A.sub.2 independently of one another represent a
hydrogen atom, a linear or branched (C.sub.1-C.sub.6)polyhaloalkyl,
a linear or branched (C.sub.1-C.sub.6)alkyl group or a cycloalkyl,
[0057] T represents a hydrogen atom, a linear or branched
(C.sub.1-C.sub.6)alkyl group optionally substituted by from one to
three halogen atoms, a group
(C.sub.1-C.sub.4)alkyl-NR.sub.1R.sub.2, or a group
(C.sub.1-C.sub.4)alkyl-OR.sub.6, [0058] R.sub.1 and R.sub.2
independently of one another represent a hydrogen atom or a linear
or branched (C.sub.1-C.sub.6)alkyl group, [0059] or R.sub.1 and
R.sub.2 form with the nitrogen atom carrying them a
heterocycloalkyl, [0060] R.sub.3 represents a linear or branched
(C.sub.1-C.sub.6)alkyl group, a linear or branched
(C.sub.2-C.sub.6)alkenyl group, a linear or branched
(C.sub.2-C.sub.6)alkynyl group, a cycloalkyl group, a
(C.sub.3-C.sub.10)cycloalkyl-(C.sub.1-C.sub.6)alkyl group wherein
the alkyl moiety is linear or branched, a heterocycloalkyl group,
an aryl group or a heteroaryl group, it being understood that one
or more of the carbon atoms of the preceding groups, or of their
possible substituents, may be deuterated, [0061] R.sub.4 represents
an aryl group, a heteroaryl group, a cycloalkyl group or a linear
or branched (C.sub.1-C.sub.6)alkyl group, it being understood that
one or more of the carbon atoms of the preceding groups, or of
their possible substituents, may be deuterated, [0062] R.sub.5
represents a hydrogen or halogen atom, a linear or branched
(C.sub.1-C.sub.6)alkyl group, or a linear or branched
(C.sub.1-C.sub.6)alkoxy group, [0063] R.sub.6 represents a hydrogen
atom or a linear or branched (C.sub.1-C.sub.6)alkyl group, [0064]
R.sub.a, R.sub.b, R.sub.c and R.sub.d, each independently of the
others, represent R.sub.7, a halogen atom, a linear or branched
(C.sub.1-C.sub.6)alkoxy group, a hydroxy group, a linear or
branched (C.sub.1-C.sub.6)polyhaloalkyl group, a trifluoromethoxy
group, --NR.sub.7R.sub.7', nitro,
R.sub.7--CO--(C.sub.0-C.sub.6)alkyl-,
R.sub.7--CO--NH--(C.sub.0-C.sub.6)alkyl-,
NR.sub.7R.sub.7'--CO--(C.sub.0-C.sub.6)alkyl-,
NR.sub.7R.sub.7'--CO--(C.sub.0-C.sub.6)alkyl-O--,
R.sub.7--SO.sub.2--NH--(C.sub.0-C.sub.6)alkyl-,
R.sub.7--NH--CO--NH--(C.sub.0-C.sub.6)alkyl-,
R.sub.7--O--CO--NH--(C.sub.0-C.sub.6)alkyl-, a heterocycloalkyl
group, or the substituents of one of the pairs (R.sub.a,R.sub.b),
(R.sub.b,R.sub.c) or (R.sub.c,R.sub.d) form together with the
carbon atoms carrying them a ring composed of from 5 to 7 ring
members, which may contain from one to 2 hetero atoms selected from
oxygen and sulphur, it also being understood that one or more
carbon atoms of the ring defined hereinbefore may be deuterated or
substituted by from one to 3 groups selected from halogen and
linear or branched (C.sub.1-C.sub.6)alkyl, [0065] R.sub.7 and
R.sub.7' independently of one another represent a hydrogen, a
linear or branched (C.sub.1-C.sub.6)alkyl, a linear or branched
(C.sub.2-C.sub.6)alkenyl, a linear or branched
(C.sub.2-C.sub.6)alkynyl, an aryl or a heteroaryl, or R.sub.7 and
R.sub.7' together with nitrogen atom carrying them form a
heterocycle composed of from 5 to 7 ring members,
[0066] it being understood that: [0067] "aryl" means a phenyl,
naphthyl, biphenyl or indenyl group, [0068] "heteroaryl" means any
mono- or bi-cyclic group composed of from 5 to 10 ring members,
having at least one aromatic moiety and containing from 1 to 4
hetero atoms selected from oxygen, sulphur and nitrogen (including
quaternary nitrogens), [0069] "cycloalkyl" means any mono- or
bi-cyclic, non-aromatic, carbocyclic group containing from 3 to 10
ring members, [0070] "heterocycloalkyl" means any mono- or
bi-cyclic, non-aromatic, condensed or spiro group composed of 3 to
10 ring members and containing from 1 to 3 hetero atoms selected
from oxygen, sulphur, SO, SO.sub.2 and nitrogen,
[0071] it being possible for the aryl, heteroaryl, cycloalkyl and
heterocycloalkyl groups so defined and the groups alkyl, alkenyl,
alkynyl and alkoxy to be substituted by from 1 to 3 groups selected
from optionally substituted, linear or branched
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.6)spiro, linear or branched,
optionally substituted (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.6)alkyl-S--, hydroxy, oxo (or N-oxide where
appropriate), nitro, cyano, --COOR', --OCOR', NR'R'', linear or
branched (C.sub.1-C.sub.6)polyhalo alkyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkylsulphonyl, halogen, optionally substituted
aryl, heteroaryl, aryloxy, arylthio, cycloalkyl, heterocycloalkyl
optionally substituted by one or more halogen atoms or alkyl
groups, it being understood that R' and R'' independently of one
another represent a hydrogen atom or an optionally substituted,
linear or branched (C.sub.1-C.sub.6)alkyl group,
[0072] it being possible for the Het group defined in formula (I')
to be substituted by from one to three groups selected from linear
or branched (C.sub.1-C.sub.6)alkyl, hydroxy, linear or branched
(C.sub.1-C.sub.6)alkoxy, NR.sub.1'R.sub.1'' and halogen, it being
understood that R.sub.1' and R.sub.1'' are as defined for the
groups R' and R'' mentioned hereinbefore,
[0073] their enantiomers and diastereoisomers, and addition salts
thereof with a pharmaceutically acceptable acid or base.
[0074] The compounds of formula (I') have pro-apoptotic properties
and as a result are of major therapeutic value in the treatment of
cancers, auto-immune diseases and diseases of the immune system. In
the present invention it has been shown that, by administering the
phosphate compounds of formula (I), the in vivo exposure to the
compounds of formula (I') was optimised. The solubility of the
compounds of formula (I) is in fact much greater than that of the
compounds of formula (I'). Consequently, using the compounds of
formula (I) in the manufacture of pharmaceutical compositions is
especially advantageous from the galenic point of view.
[0075] The invention relates also to a process for the preparation
of compounds of formula (I), which process is characterised in that
there is used as starting material the compound of formula
(II):
##STR00005##
[0076] wherein R.sub.a, R.sub.b, R.sub.c and R.sub.d are as defined
for formula (I'),
[0077] which compound of formula (II) is subjected to a Heck
reaction, in an aqueous or organic medium, in the presence of a
palladium catalyst, of a base, of a phosphine and of the compound
of formula (III):
##STR00006##
[0078] wherein the groups A.sub.1, A.sub.2, X and Y are as defined
for formula (I') and Alk represents a linear or branched
(C.sub.1-C.sub.6)alkyl,
[0079] to obtain the compound of formula (IV):
##STR00007##
[0080] wherein A.sub.1, A.sub.2, X, Y, R.sub.a, R.sub.b, R.sub.c
and R.sub.d are as defined for formula (I') and Alk is as defined
hereinbefore,
[0081] the aldehyde function of which compound of formula (IV) is
oxidised to a carboxylic acid to form the compound of formula
(V):
##STR00008##
[0082] wherein A.sub.1, A.sub.2, X, Y, R.sub.a, R.sub.b, R.sub.c
and R.sub.d are as defined for formula (I') and Alk is as defined
hereinbefore,
[0083] which compound of formula (V) is then subjected to peptide
coupling with a compound of formula (VI):
##STR00009##
[0084] wherein T and R.sub.5 are as defined for formula (I'),
[0085] to yield the compound of formula (VII):
##STR00010##
[0086] wherein A.sub.1, A.sub.2, X, Y, R.sub.a, R.sub.b, R.sub.c,
R.sub.d, T and R.sub.5 are as defined for formula (I') and Alk is
as defined hereinbefore,
[0087] the ester function of which compound of formula (VII) is
hydrolysed to yield the corresponding carboxylic acid or
carboxylate, which may be converted into an acid derivative such as
the corresponding acyl chloride or anhydride before being coupled
with an amine NHR.sub.3R.sub.4 wherein R.sub.3 and R.sub.4 have the
same meanings as for formula (I'), before being subjected to the
action of a pyrophosphate, phosphonate or phosphoryl compound under
basic conditions, it being possible for the compound thereby
obtained to be optionally hydrolysed or hydrogenolysed to yield the
compound of formula (I),
[0088] which compound of formula (I) may be purified according to a
conventional separation technique, which is converted, if desired,
into its addition salts with a pharmaceutically acceptable acid or
base and which is optionally separated into its isomers according
to a conventional separation technique,
[0089] it being understood that, at any time considered appropriate
in the course of the above-described process, certain groups
(hydroxy, amino . . . ) of the reagents or intermediates of
synthesis may be protected and then deprotected according to the
requirements of synthesis.
[0090] The compounds of formulae (II), (III), (VI) and the amine
NHR.sub.3R.sub.4 are either commercially available or can be
obtained by the person skilled in the art using conventional
chemical reactions described in the literature.
[0091] More specifically, the phosphate compounds of formula (I)
according to the invention will be useful in the treatment of
chemo- or radio-resistant cancers and also in malignant
haemopathies and small-cell lung cancer.
[0092] Among the cancer treatments envisaged there may be
mentioned, without implying any limitation, cancers of the bladder,
brain, breast and uterus, chronic lymphoid leukaemias, colorectal
cancer, cancers of the oesophagus and liver, lymphoblastic
leukaemias, non-Hodgkin lymphomas, melanomas, malignant
haemopathies, myelomas, ovarian cancer, non-small-cell lung cancer,
prostate cancer and small-cell lung cancer. Among non-Hodgkin
lymphomas, there may be mentioned more preferably follicular
lymphomas, mantle cell lymphomas, diffuse large B-cell lymphomas,
small lymphocytic lymphomas and marginal zone B-cell lymphomas.
[0093] The present invention relates also to pharmaceutical
compositions comprising at least one compound of formula (I) in
combination with one or more pharmaceutically acceptable
excipients.
[0094] Among the pharmaceutical compositions according to the
invention there may be mentioned more especially those that are
suitable for oral, parenteral, nasal, per- or trans-cutaneous,
rectal, perlingual, ocular or respiratory administration,
especially tablets or dragees, sublingual tablets, sachets,
paquets, capsules, glossettes, lozenges, suppositories, creams,
ointments, dermal gels, and drinkable or injectable ampoules.
[0095] The dosage varies according to the sex, age and weight of
the patient, the administration route, the nature of the
therapeutic indication, or of any associated treatments, and ranges
from 0.01 mg to 1 g per 24 hours in one or more
administrations.
[0096] Furthermore, the present invention relates also to the
association of a compound of formula (I) with an anticancer agent
selected from genotoxic agents, mitotic poisons, anti-metabolites,
proteasome inhibitors, kinase inhibitors and antibodies, and also
to pharmaceutical compositions comprising that type of association
and their use in the manufacture of medicaments for use in the
treatment of cancer.
[0097] The compounds of the invention may also be used in
association with radiotherapy in the treatment of cancer.
[0098] The following Preparations and Examples illustrate the
invention without limiting it in any way.
Preparation 1:
6-[1-(Methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-1,3-benzodioxole-
-5-carboxylic acid
Step A: 1-Formyl-2-piperidine-carboxylic acid
[0099] To a solution of 40 g of a racemic mixture of
2-piperidine-carboxylic acid (0.310 mmol) in 300 mL of formic acid
placed at 0.degree. C. there are added, dropwise, 200 mL (2.15
mmol) of acetic anhydride. The batch is then stirred at ambient
temperature overnight. Then, the reaction mixture is cooled to
0.degree. C., hydrolysed by adding 250 mL of water, and stirred for
half an hour at 0.degree. C. before being concentrated to dryness.
The oil thereby obtained is taken up in 200 mL of methanol and then
concentrated to dryness. The title product is obtained in the form
of an oil in a yield of 98%. It is used directly, without being
otherwise purified, in the next Step.
[0100] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300.degree. K): 13.0
(m, 1H OH); 8.0-8.05 (2s, 1H aldehyde); 4.9-4.5 (2d, 1H .alpha. to
the N and COOH); 4.1-2.6 (m, 2H .alpha. to the N); 2.2-1.2 (m, 6H
piperidine)
[0101] IR: .nu.: --OH: 2000-3000 cm.sup.-1 acid; .nu.: >C.dbd.O
1703 cm.sup.-1 wide band
Step B: Methyl 5,6,7,8-tetrahydro-1-indolizine-carboxylate
[0102] To a solution of 10 g of the carboxylic acid obtained in
Step A (63.6 mmol) in 65 mL of dichloroethane there are
successively added 13.4 g of tosyl chloride (70.4 mmol), 11.5 mL of
methyl 2-chloroacrylate (113.5 mmol) and then, dropwise, 17.8 mL of
N,N,N-triethylamine (127.2 mmol). The reaction mixture is then
refluxed for 1 hour 30 minutes. It is then placed at ambient
temperature, and there are then added 5 mL of methyl
2-chloroacrylate (48.9 mmol) and, dropwise, 9 mL of
N,N,N-triethylamine (64 mmol). The batch is refluxed overnight. The
reaction mixture is then diluted with methylene chloride, washed
successively with 1M HCl solution, saturated aqueous NaHCO.sub.3
solution and then with brine until a neutral pH is obtained. The
organic phase is then dried over MgSO.sub.4, filtered, concentrated
to dryness and purified by chromatography over silica gel
(heptane/AcOEt gradient). The title product is obtained in the form
of an oil.
[0103] .sup.1H NMR: .delta. (400 MHz; CDCl.sub.3; 300.degree. K):
6.55-6.40 (d, 2H, tetrahydroindolizine); 3.91 (t, 3H methyl ester);
3.78 (s, 3H tetrahydroindolizine); 3.08 (t, 2H,
tetrahydroindolizine); 1.95-1.85 (m, 4H, tetrahydroindolizine)
[0104] IR: .nu.: >C.dbd.O 1692 cm.sup.-1 ester
Step C: Methyl
3-(6-formyl-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydro-1-indolizine-carboxy-
late
[0105] To a solution of 6.4 g of the ester obtained in Step B (35.7
mmol) in 12 mL of N,N-dimethylacetamide, there are successively
added 12.3 g of 6-bromo-1,3-benzodioxole-5-carbaldehyde (53.6 mmol)
and 7 g of potassium acetate (71.4 mmol), and then the batch is
stirred under argon for 20 minutes. There are then added 1.3 g of
palladium catalyst dichlorobis(triphenylphosphine)palladium(II)
(PdCl.sub.2(PPh.sub.3).sub.2) (1.8 mmol). The reaction mixture is
then heated at 130.degree. C. for one hour before adding 139 .mu.L
of H.sub.2O thereto. Heating is maintained at that same temperature
overnight. The mixture is allowed to return to ambient temperature
and it is then diluted with AcOEt. Animal charcoal (25 g per g of
product) is added and the batch is stirred at ambient temperature
for 1 hour and then filtered. The organic phase is then washed with
water, dried over magnesium sulphate and concentrated to dryness.
The crude product thereby obtained is purified by chromatography
over silica gel (heptane/AcOEt gradient). The title product is
obtained in the form of an oil.
[0106] .sup.1H NMR: .delta.: (400 MHz; dmso-d6; 353.degree. K):
9.65 (s, 1H, H aldehyde); 7.3-7.15 (2s, 2H, aromatic Hs); 6.45 (s,
1H tetrahydroindolizine); 6.20 (s, 2H methylenedioxy); 3.70 (s, 3H
methyl ester); 3.5-4.0 (m, 2H tetrahydroindolizine); 3.05 (m, 2H
tetrahydroindolizine); 1.85 (m, 4H tetrahydroindolizine) IR: .nu.:
>C.dbd.O 1695 cm.sup.-1 ester; .nu.: >C.dbd.O 1674
cm.sup.-1
Step D:
6-[1-(Methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-1,3-benzo-
dioxole-5-carboxylic acid
[0107] A solution containing 3.37 g of the compound obtained in
Step C (10.3 mmol) in 9.3 mL of acetone and 8.8 mL (80.24 mmol) of
2-methyl-2-butene is prepared and placed at 0.degree. C. There are
added, dropwise, 9.3 mL of an aqueous solution containing a mixture
of 3.3 g of sodium chlorite (NaClO.sub.2) (36.05 mmol) and 3.6 g of
sodium dihydrogen phosphate monohydrate (NaH.sub.2PO.sub.4) (25.75
mmol). The batch is then stirred at ambient temperature for 7
hours. The reaction mixture is then concentrated in order to remove
the acetone. The solid then obtained is filtered off, washed with
water and then dried at 40.degree. C. in vacuo overnight. The title
product is obtained in the form of a solid, which is subsequently
used without being otherwise purified.
[0108] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300.degree. K):
12.10 (m, 1H, H carboxylic acid); 7.40-6.88 (2s, 2H, aromatic Hs);
6.20 (s, 1H, H tetrahydroindolizine); 6.18 (s, 2H, H
methylenedioxy); 3.70 (s, 3H, methyl ester); 3.55 (t, 2H
tetrahydroindolizine); 3.00 (t, 2H tetrahydroindolizine); 1.80 (m,
4H, H tetrahydroindolizine)
[0109] IR: .nu.: --OH: 3000-2000 cm.sup.-1 acid; .nu.: >C.dbd.O
1686-1676 cm.sup.-1 ester+acid; .nu.: >C.dbd.C< 1608
cm.sup.-1
Preparation 2:
2-[1-(Methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]benzoic
acid
[0110] The procedure is in accordance with the protocol described
in Preparation 1, replacing the
6-bromo-1,3-benzodioxole-5-carbaldehyde used in Step C by
2-bromo-benzaldehyde.
Preparation 3:
6-[1-(Methoxycarbonyl)-3-indolizinyl]-1,3-benzodioxole-5-carboxylic
acid
Step A: 1-(Carboxymethyl)-1,2-dihydropyridinium bromide
[0111] To a solution of 16.2 mL of pyridine (200 mmol) in 120 mL of
ethyl acetate there are added, in portions, 27.8 g (200 mmoles) of
bromoacetic acid. The batch is then stirred at ambient temperature
overnight. The precipitate thereby obtained is filtered off and
then washed with cold ethyl acetate. After drying, the title
product is obtained in the form of a powder which is used directly
in the next Step.
[0112] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300.degree. K): 9.15
(d, 2H, aromatic Hs pyridine); 8.7 (t, 1H, aromatic H); 8.25 (t,
2H, aromatic H); 5.65 (s, 2H, H CH.sub.2COOH)
[0113] IR: .nu.: C.dbd.O: 1732 cm.sup.-1; --OH acid: 2800
cm.sup.-1
Step B: Methyl 1-indolizinecarboxylate
[0114] To a suspension of 6.55 g of the pyridinium salt obtained in
Step A (30 mmol) in 240 mL of toluene there are successively added
16.7 mL of methyl acrylate (150 mmol), 4.2 mL of triethylamine (30
mmol) and then, in portions, 20.9 g of MnO.sub.2 (240 mmol). The
batch is then heated at 90.degree. C. for 3 hours. After cooling,
the reaction mixture is filtered over a cake of Celite and
concentrated to dryness. The title product is then isolated by
purification over silica gel (heptane/AcOEt gradient: 0-10%) in the
form of an oil which crystallises in the cold state.
[0115] .sup.1H NMR: .delta. (300 MHz; dmso-d6; 300.degree. K): 8.5
(d, 1H, H indolizine); 8.05 (d, 1H, H indolizine); 7.6 (s, 1H, H
indolizine); 7.15 (m, 2H, H indolizine); 6.85 (m, 1H, H
indolizine); 4.25 (q, 2H, --C(O)CH.sub.2CH.sub.3); 1.35 (t, 3H,
--C(O)CH.sub.2CH.sub.3)
[0116] IR: .nu.: C.dbd.O ester: 1675 cm.sup.-1; aromatic C.dbd.C
moieties: 1634 cm.sup.-1
Step C:
6-[1-(Methoxycarbonyl)-3-indolizinyl]-1,3-benzodioxole-5-carboxyli-
c acid
[0117] The procedure is in accordance with the protocol described
in Steps C and D of Preparation 1.
Preparation 4:
4-Chloro-2-[4-(ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]-benzoic
acid
Step A: Ethyl 1,2-dimethyl-1H-pyrrole-3-carboxylate
[0118] To a solution of 10 g of ethyl
2-methyl-1H-pyrrole-3-carboxylate (65.3 mmol) and 8.95 mL (130.6
mmol) of methyl iodide in 70 mL of dimethylformamide placed at
0.degree. C. there are added, in three portions, 2.61 g (65.3 mmol)
of sodium hydride 60%. The batch is then stirred at 0.degree. C.
for 1 hour. Then, the reaction mixture is hydrolysed by the
addition of 420 mL of ice-cold water. The reaction mixture is then
diluted with ethyl acetate, successively washed with 0.1M HCl
solution, saturated aqueous LiCl solution and then brine. The
organic phase is then dried over MgSO.sub.4, filtered, concentrated
to dryness and purified by chromatography over silica gel
(petroleum ether/AcOEt gradient).
[0119] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300K): 6.65 (d, 1H
pyrrole); 6.3 (1d, 1H pyrrole); 4.1 (1q, 2H, OCH.sub.2CH.sub.3);
3.5 (s, 3H N-pyrrole); 2.4 (s, 3H pyrrole); 1.5 (1t, 3H
OCH.sub.2CH.sub.3)
[0120] IR: .nu.: >C.dbd.O: 1688 cm.sup.-1; .nu.: C--O--C: 1172
cm.sup.-1
Step B: Ethyl
5-(5-chloro-2-formylphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxylate
[0121] To a solution of 10.5 g of the compound obtained in Step A
(62.8 mmol) in 65 mL of N,N-dimethylacetamide there are
successively added 15.2 g of 2-bromo-4-chlorobenzaldehyde (69
mmol), 12.3 g of potassium acetate (125.6 mmol) and then the batch
is stirred under argon for 20 minutes. There are then added 2.2 g
of palladium catalyst PdCl.sub.2(PPh.sub.3).sub.2 (3.14 mmol). The
reaction mixture is then heated at 130.degree. C. overnight. The
mixture is allowed to return to ambient temperature and it is then
diluted with dichloromethane. Animal charcoal is added (30 g) and
the batch is stirred at ambient temperature for 1 hour and then
filtered. The organic phase is then washed with water, dried over
magnesium sulphate and concentrated to dryness. The crude product
thereby obtained is purified by chromatography over silica gel
(petroleum ether/AcOEt gradient). The title product is obtained in
the form of a solid.
[0122] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300K): 9.8 (s, 1H,
formyl); 7.91-7.69-7.61 (d, 3H, aromatic Hs); 6.5 (s, 1H pyrrole);
4.2 (q, 2H, OCH.sub.2CH.sub.3); 3.4 (s, 3H, CH.sub.3--N-pyrrole);
2.55 (s, 3H pyrrole); 1.28 (t, 3H, OCH.sub.2CH.sub.3)
Step C:
4-Chloro-2-[4-(ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]benzoic
acid
[0123] A solution is prepared containing 12.85 g of the compound
obtained in Step B (42 mmol) and 35.7 mL (336 mmol) of
2-methyl-2-butene in a mixture containing 20 mL of acetone and 20
mL of tetrahydrofuran. There are added, dropwise, 200 mL of an
aqueous solution containing a mixture of 13.3 g of sodium chlorite
(NaClO.sub.2) (147 mmol) and 14.5 g of sodium dihydrogen phosphate
monohydrate (NaH.sub.2PO.sub.4H.sub.2O) (105 mmol). The batch is
then vigorously stirred at ambient temperature for 7 hours. The
reaction mixture is then concentrated to remove the acetone. Ethyl
acetate is added, and the organic phase is washed with water and
then concentrated to dryness. The residue is then taken up in a
minimum of ethyl ether. The solid then obtained is filtered off,
washed with ether and then dried in vacuo at 40.degree. C.
overnight. The title product is obtained in the form of a solid,
which is subsequently used without being otherwise purified.
[0124] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300K): 13 (m, 1H
COOH); 7.85-7.6-7.41(d,dd, wd, 3H, aromatic Hs); 6.3 (s, 1H, H
pyrrole); 4.15 (q, 2H, OCH.sub.2CH.sub.3); 3.25 (s, 3H,
CH.sub.3--N-pyrrole); 2.5 (s, 3H, CH.sub.3-pyrrole); 1.25 (t, 3H,
OCH.sub.2CH.sub.3)
[0125] IR: .nu.: --OH: 3100-2500 cm.sup.-1 acid; .nu.: >C.dbd.O:
1681 cm.sup.-1 ester+acid
Preparation 5:
6-[4-(Ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]-1,3-benzodioxole-5-car-
boxylic acid
[0126] The procedure is in accordance with the process of
Preparation 4, replacing the 2-bromo-4-chlorobenzaldehyde used in
Step B by 6-bromo-1,3-benzodioxole-5-carbaldehyde.
Preparation 6:
4-Fluoro-3-methoxy-2-[4-(ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]benz-
oic acid
[0127] The procedure is in accordance with the process of
Preparation 4, replacing the 2-bromo-4-chlorobenzaldehyde used in
Step B by 2-bromo-4-fluoro-3-methoxybenzaldehyde.
Preparation 7:
4-Fluoro-2-[4-(ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]benzoic
acid
[0128] The procedure is in accordance with the process of
Preparation 4, replacing the 2-bromo-4-chlorobenzaldehyde used in
Step B by 2-bromo-4-fluorobenzaldehyde.
Preparation 8:
7-[4-(Methoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]-2,3-dihydro-1,4-benzo-
dioxin-6-carboxylic acid
[0129] The procedure is in accordance with the process of
Preparation 4, replacing the ethyl
2-methyl-1H-pyrrole-3-carboxylate in Step A by methyl
2-methyl-1H-pyrrole-3-carboxylate and the
2-bromo-4-chlorobenzaldehyde used in Step B by
7-bromo-2,3-dihydro-1,4-benzodioxin-6-carbaldehyde.
Preparation 9:
5-Benzyloxy-2-(1-methoxycarbonyl-5,6,7,8-tetrahydroindolizin-3-yl)benzoic
acid
Step A: Methyl
3-(4-benzyloxy-2-formyl-phenyl)-5,6,7,8-tetrahydroindolizine-1-carboxylat-
e
[0130] 5-Benzyloxy-2-bromo-benzaldehyde (12.3 g, 42.2 mmol) is
introduced into a flask in the presence of potassium acetate (8.3
g; 84.2 mmol) and 120 mL of dimethylacetamide. After degassing
under argon, dichlorobis(triphenylphosphine)palladium (II) (1.04 g,
1.5 mmol) is added and the mixture is then degassed under argon
before being heated at 100.degree. C. for 16 hours. After returning
to ambient temperature, the reaction mixture is poured into 200 mL
of ethyl acetate, filtered over Celite, and washed with water and
then with brine. The combined aqueous phases are extracted with
ethyl acetate. The organic phases are dried over sodium sulphate,
filtered and concentrated under reduced pressure. The residue
obtained is purified by chromatography over silica gel in order to
obtain the title product.
Step B:
5-Benzyloxy-2-(1-methoxycarbonyl-5,6,7,8-tetrahydroindolizin-3-yl)-
benzoic acid
[0131] To a solution of the compound obtained in Step B (4.63 g,
11.89 mmol) in 300 mL of acetone there is added 2-methyl-2-butene
(6.31 mL, 59 mmol). A solution of sodium dihydrogen phosphate
monohydrate (6.56 g, 47.6 mmol) and sodium chlorite (2.69 g, 23.8
mmol) in 40 mL of water is then poured in dropwise whilst
maintaining the temperature below 20.degree. C. After stirring for
30 minutes at ambient temperature, the mixture is acidified with 2M
HCl solution and then the phases are separated. The aqueous phase
is extracted with ethyl acetate. The combined organic phases are
dried over sodium sulphate, filtered and evaporated to dryness to
provide the expected compound.
Preparation 1':
(3S)-3-(4-Morpholinylmethyl)-1,2,3,4-tetrahydroisoquinoline
Step A: Benzyl
(3S)-3-(4-morpholinylcarbonyl)-3,4-dihydro-2(1H)-isoquinoline-carboxylate
[0132] To a solution of 5 g of
(3S)-2-[(benzyloxy)carbonyl]-1,2,3,4-tetrahydro-3-isoquinolinecarboxylic
acid (16 mmol) in 160 mL of dichloromethane there are added 1.5 mL
of morpholine (17.6 mmol), then 9 mL of N,N,N-triethylamine (64
mmol), 3.3 g of 1-ethyl-3-(3'-dimethylaminopropyl)-carbodiimide
(EDC) (19.2 mmol) and 2.6 g of hydroxybenzotriazole (HOBt) (19.2
mmol). The reaction mixture is stirred at ambient temperature
overnight; it is then poured into aqueous ammonium chloride
solution and extracted with ethyl acetate. The organic phase is
then dried over magnesium sulphate, and then filtered and
evaporated to dryness. The crude product thereby obtained is then
purified by chromatography over silica gel
(dichloromethane/methanol gradient). The product is obtained in the
form of a foam.
[0133] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 353K): 7.30 (m, 5H
benzyl); 7.15 (m, 4H, aromatic Hs); 5.2-5.0 (m, 3H, 2H benzyl, 1H
dihydroisoquinoline); 4.75-4.5 (2d, 2H dihydroisoquinoline);
3.55-3.3 (m, 8H morpholine); 3.15-2.9 (2dd, 2H
dihydroisoquinoline)
[0134] IR: .nu.: >C.dbd.O: 1694; 1650 cm.sup.-1
Step B: Benzyl
(3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinecarboxylate
[0135] To a solution of 5.3 g of the product obtained in Step A
(13.9 mmol) in 278 mL of tetrahydrofuran there are added 14 mL of
borane-dimethylsulphide complex (BH.sub.3Me.sub.2S) (27.8 mmol) at
ambient temperature. The batch is heated for 4 hours at 80.degree.
C. It is allowed to return to ambient temperature and there are
then added 7 mL (14 mmol) of BH.sub.3Me.sub.2S. The reaction
mixture is again heated at 80.degree. C. for 2 hours. The
tetrahydrofuran is then evaporated off and then there is slowly
added methanol and then 5.6 mL of 5M hydrochloric acid (27.8 mmol).
The mixture is stirred at ambient temperature overnight, and then
at 80.degree. C. for 1 hour. Saturated aqueous NaHCO.sub.3 solution
is then added to the reaction mixture placed at 0.degree. C. until
a pH of 8 is obtained, and extraction with ethyl acetate is then
carried out. The organic phase is then dried over magnesium
sulphate, and then filtered and evaporated to dryness. The title
product is obtained in the form of an oil.
[0136] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 353K): 7.43-7.30
(unresolved peak, 5H benzyl); 7.19 (m, 4H, aromatic Hs); 5.16 (m,
2H, 2H benzyl); 4.79-4.29 (d, 2H dihydroisoquinoline); 4.58 (m, 1H
dihydroisoquinoline); 3.50 (m, 4H morpholine); 3.02-2.80 (dd, 2H
dihydroisoquinoline); 2.42-2.28 (unresolved peak, 5H, 4H
morpholine, 1H morpholine); 2.15 (dd, 1H morpholine)
[0137] IR: .nu.: >CH: 2810 cm.sup.-1; .nu.: >C.dbd.O: 1694
cm.sup.-1; .nu.: >C--O--C<: 1114 cm.sup.-1; .nu.: >CH--Ar:
751; 697 cm.sup.-1
Step C:
(3S)-3-(4-Morpholinylmethyl)-1,2,3,4-tetrahydroisoquinoline
[0138] To a solution of 4.9 g of the compound of Step B (13.4 mmol)
in 67 mL of ethanol there is added 0.980 g of palladium dihydroxide
(20% by weight) at ambient temperature. The reaction mixture is
placed under 1.2 bars of hydrogen at ambient temperature for 4
hours. It is then passed through a Whatman filter and the palladium
is then rinsed several times with ethanol. The filtrate is
evaporated to dryness. The title product is obtained in the form of
an oil.
[0139] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300K): 7.12-7.0
(unresolved peak, 4H, aromatic Hs); 3.92 (s, 2H
tetrahydroisoquinoline); 3.60 (t, 4H morpholine); 2.98 (m, 1H
tetrahydroisoquinoline); 2.68 (dd, 1H tetrahydroisoquinoline);
2.5-2.3 (unresolved peak, 8H, 1H tetrahydroisoquinoline, 6H
morpholine, 1H NH)
[0140] IR: .nu.: >NH: 3322 cm.sup.-1; .nu.: >C--O--C<:
1115 cm.sup.-1; .nu.: >CH--Ar: 742 cm.sup.-1
Preparation 2': (3R)-3-Methyl-1,2,3,4-tetrahydroisoquinoline
hydrochloride
Step A:
{(3S)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin--
3-yl}methyl 4-methylbenzenesulphonate
[0141] To a solution of 30.2 g of
[(3S)-1,2,3,4-tetrahydroisoquinolin-3-yl]methanol (185 mmol) in 750
mL of dichloromethane there are successively added 91.71 g of tosyl
chloride (481 mmol) and then, dropwise, 122.3 mL of
N,N,N-triethylamine (740 mmol). The reaction mixture is then
stirred at ambient temperature for 20 hours. It is then diluted
with dichloromethane, washed successively with 1M HCl solution,
saturated aqueous NaHCO.sub.3 solution and then brine until
neutral. The organic phase is then dried over MgSO.sub.4, filtered
and concentrated to dryness. The solid obtained is then dissolved
in a minimum volume of dichloromethane and then cyclohexane is
added until a precipitate is formed. This precipitate is then
filtered off and washed with cyclohexane. After drying, the title
product is obtained in the form of crystals.
[0142] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300.degree. K): 7.75
(d, 2H, aromatic Hs, ortho O-tosyl); 7.6 (d, 2H, aromatic Hs, ortho
N-tosyl); 7.5 (d, 2H, aromatic Hs, meta O-tosyl); 7.3 (d, 2H,
aromatic Hs, meta N-tosyl); 7.15-6.9 (m, 4H, aromatic Hs,
tetrahydroisoquinoline); 4.4-4.15 (dd, 2H, aliphatic Hs,
tetrahydroisoquinoline); 4.25 (m, 1H, aliphatic H,
tetrahydroisoquinoline); 4.0-3.8 (2dd, 2H, aliphatic Hs,
CH.sub.2--O-tosyl); 2.7 (2dd, 2H, aliphatic Hs,
tetrahydroisoquinoline); 2.45 (s, 3H, O--SO.sub.2-Ph-CH.sub.3);
2.35 (s, 3H, N--SO.sub.2-Ph-CH.sub.3)
[0143] IR: .nu.: --SO.sub.2: 1339-1165 cm.sup.-1
Step B:
(3R)-3-Methyl-2-[(4-methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoq-
uinoline
[0144] To a suspension of 8.15 g (214.8 mmol) of LiAlH.sub.4 in 800
mL of methyl tert-butyl ether (MTBE) there are added 101.2 g of the
ditosyl compound obtained in Step A (214.8 mmol) dissolved in 200
mL of MTBE. The batch is then heated at 50.degree. C. for 2 hours.
It is allowed to cool and placed at 0.degree. C., and there are
then added, dropwise, 12 mL of 5M NaOH solution. The batch is
stirred at ambient temperature for 45 minutes. The solid thereby
obtained is then filtered off and washed with MTBE and then with
dichloromethane. The filtrate is then concentrated to dryness. The
title product is then obtained in the form of a solid.
[0145] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300.degree. K): 7.70
(d, 2H, aromatic Hs, ortho N-tosyl); 7.38 (d, 2H, aromatic Hs, meta
N-tosyl); 7.2-7.0 (m, 4H, aromatic Hs, tetrahydroisoquinoline); 4.4
(m, 2H, aliphatic Hs, tetrahydroisoquinoline); 4.3 (m, 1H,
aliphatic H, tetrahydroisoquinoline); 2.85-2.51 (2dd, 2H, aliphatic
Hs, tetrahydroisoquinoline); 2.35 (s, 3H, N--SO.sub.2-Ph-CH.sub.3);
0.90 (d, 3H, tetrahydroisoquinoline-CH.sub.3)
[0146] IR: .nu.: --SO.sub.2: 1332-1154 cm.sup.-1
Step C: (3R)-3-Methyl-1,2,3,4-tetrahydroisoquinoline
[0147] To a solution of 31.15 g (103.15 mmol) of the monotosyl
compound obtained in Step B in 500 mL of anhydrous methanol there
are added, in portions, 3.92 g (161 mmol) of magnesium turnings.
The batch is stirred in the presence of ultrasound for 96 hours.
The reaction mixture is then filtered and the solid is washed
several times with methanol. The filtrate is then concentrated to
dryness. After purification by chromatography over silica gel
(dichloromethane/EtOH/NH.sub.4OH gradient), the title product is
obtained in the form of an oil.
[0148] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300.degree. K): 7.05
(m, 4H, aromatic Hs, tetrahydroisoquinoline); 3.90 (m, 2H,
aliphatic Hs, tetrahydroisoquinoline); 2.85 (m, 1H, aliphatic H,
tetrahydroisoquinoline); 2.68-2.4 (2dd, 2H, aliphatic Hs,
tetrahydroisoquinoline); 1.12 (d, 3H,
tetrahydroisoquinoline-CH.sub.3); 2.9-2.3 (m, broad, 1H, HN
(tetrahydroisoquinoline))
[0149] IR: .nu.: --NH: 3248 cm.sup.-1
Step D: (3R)-3-Methyl-1,2,3,4-tetrahydroisoquinoline
hydrochloride
[0150] To a solution of 14.3 g (97.20 mmol) of the compound
obtained in Step C in 20 mL of anhydrous ethanol there are added,
dropwise, 100 mL of a 1M solution of HCl in ether. The batch is
stirred at ambient temperature for 1 hour and then filtered. The
crystals thereby obtained are washed with ethyl ether. After
drying, the title product is obtained in the form of crystals.
[0151] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300.degree. K): 9.57
(m, broad, 2H, NH.sub.2.sup.+ (tetrahydroisoquinoline); 7.22 (m,
4H, aromatic Hs, tetrahydroisoquinoline); 4.27 (s, 2H, aliphatic
Hs, tetrahydroisoquinoline); 3.52 (m, 1H, aliphatic H,
tetrahydroisoquinoline); 3.03-2.85 (2dd, 2H, aliphatic Hs,
tetrahydroisoquinoline); 1.39 (d, 3H,
tetrahydroisoquinoline-CH.sub.3)
[0152] IR: .nu.: --NH.sub.2.sup.+: 3000-2300 cm.sup.-1; .nu.:
aromatic --CH: 766 cm.sup.-1
Preparation 3':
(3R)-3-[3-(Morpholin-4-yl)propyl]-1,2,3,4-tetrahydroisoquinoline
Step A:
{(3S)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin--
3-yl}methyl 4-methylbenzenesulphonate
[0153] The procedure is the same as that of Step A of Preparation
2'.
Step B: tert-Butyl
2-({(3R)-2-[(4-methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl-
}methyl)-3-(morpholin-4-yl)-3-oxopropanoate
[0154] To a suspension of 1 g of NaH (60%) (25.08 mmol) in 30 mL of
MTBE there are added, dropwise, a solution of 5 g of tert-butyl
3-morpholino-3-oxopropanoate (21.81 mmol) in 20 mL of anhydrous
MTBE. This suspension is stirred at ambient temperature for 1 hour
and then the compound obtained in Step A is added in the form of a
powder. The batch is stirred at 60.degree. C. for 30 hours. 100 mL
of saturated aqueous ammonium chloride solution are added. The
resulting solution is extracted with dichloromethane. The organic
phase is then dried over MgSO.sub.4, filtered and concentrated to
dryness. After purification by chromatography over silica gel
(dichloromethane/MeOH gradient), the expected product is obtained
in the form of an oil.
[0155] .sup.1H NMR (500 MHz, dmso-d6) .delta. ppm: 7.63/7.59 (2d,
2H), 7.3/7.26 (2d, 2H), 7.13 (m, 2H), 7.09/6.97 (2t, 2H),
4.64/4.55/4.36/4.28 (2AB, 2H), 4.25/4.11 (2m, 1H), 3.81 (m, 1H),
3.73-3.48 (m, 4H), 3.57-3.32 (m, 4H), 2.51 (m, 2H), 2.32/2.31 (2s,
3H), 1.88/1.79 (2m, 2H), 1.39/1.38 (2s, 9H)
[0156] IR (ATR) cm.sup.-1: .nu.: >C.dbd.O: 1731 (ester); .nu.:
>C.dbd.O: 1644 (amide); .nu.: --SO2: 1334-1156; .nu.:
>C--O--C<: 1115; .gamma.: >CH--Ar: 815-746-709
Step C:
2-({(3R)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinol-
in-3-yl}methyl)-3-(morpholin-4-yl)-3-oxopropanoic acid
[0157] To a solution of 9.5 g (17.97 mmol) of the compound obtained
in Step B in 40 mL of dioxane there are added, dropwise, 20 mL of a
4M solution of HCl in dioxane. The batch is stirred at ambient
temperature for 48 hours and then the solution is concentrated to
dryness. After drying, the expected product is obtained in the form
of an oil.
[0158] .sup.1H NMR (400 MHz, dmso-d6) .delta. ppm: 12.75 (m, 1H),
7.6 (2*d, 2H), 7.3 (2*d, 2H), 7.1/6.95 (2*m, 4H), 4.7-4.2 (d, 2H),
4.25/4.12 (2*m, 1H), 3.9-3.3 (m, 9H), 2.55 (d, 2H), 2.3 (2*s, 3H),
1.8 (t, 2H)
[0159] IR (ATR) cm.sup.-1: .nu.: --OH : 3500 to 2000; .nu.:
>C.dbd.O: 1727 (acid); .nu.: >C.dbd.O: 1634 (amide); .nu.:
--SO2: 1330-1155
Step D:
3-{(3R)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinoli-
n-3-yl}-1-(morpholin-4-yl)propan-1-one
[0160] To a solution of 7.80 g (16.51 mmol) of the compound
obtained in Step C in 100 mL of DMSO there are added 1.16 g (19.83
mmol) of solid sodium chloride and then, dropwise, 5 mL of water.
The batch is stirred at 130.degree. C. for 1 hour and then the
solution is concentrated to 3/4. The reaction mixture is then
diluted with dichloromethane and washed successively with saturated
aqueous lithium chloride solution and then with brine. The organic
phase is then dried over MgSO.sub.4, filtered and concentrated to
dryness. After purification by chromatography over silica gel
(cyclohexane/ethyl acetate gradient), the expected product is
obtained in the form of an oil.
[0161] .sup.1H NMR (400 MHz, dmso-d6) .delta. ppm: 7.65 (d, 2H),
7.3 (d, 2H), 7.15/7 (2 m, 4H), 4.6 (d, 1H), 4.25 (d, 1H), 4.2 (m,
1H), 3.5 (m, 4H), 3.4 (2 m, 4H), 2.6 (2 dd, 2H), 2.35 (s, 3H), 2.3
(m, 2H), 1.5 (quad., 2H)
[0162] IR (ATR) cm.sup.-1: .nu.: >C.dbd.O: 1639; .nu.: --SO2:
1331-1156; .gamma.: >CH--Ar: 815-675
Step E:
(3R)-2-[(4-Methylphenyl)sulphonyl]-3-[3-(morpholin-4-yl)propyl]-1,-
2,3,4-tetrahydroisoquinoline
[0163] To a solution of 6.0 g (14.0 mmol) of the compound obtained
in Step D in 60 mL of MTBE and 14 mL of dichloromethane there are
added 1.06 g (28 mmol) of LAH in portions over 5 minutes. The batch
is stirred at ambient temperature for 15 hours. There are added,
dropwise, 1.5 mL of water and stirring is carried out for 15
minutes. There are then added, dropwise, 1.5 mL of 5M sodium
hydroxide solution and stirring is carried out for 15 minutes. The
reaction mixture is then diluted with MTBE and dichloromethane. The
suspension is then filtered and the precipitate is washed with MTBE
and dichloromethane. The organic phase is then dried over
MgSO.sub.4, filtered and concentrated to dryness. After
purification by chromatography over silica gel
(dichloromethane/EtOH/NH.sub.4OH gradient), the expected product is
obtained in the form of an oil.
[0164] .sup.1H NMR (400 MHz, dmso-d6) .delta. ppm: 7.68 (d, 2H),
7.32 (d, 2H), 7.1 (unresolved peak, 4H), 4.65/4.23 (AB, 2H), 4.2
(m, 1H), 3.55 (t, 4H), 2.7/2.6 (ABx, 2H), 2.35 (s, 3H), 2.25 (t,
4H), 2.2 (t, 2H), 1.4/1.3 (2m, 4H).
[0165] IR (ATR) cm.sup.-1: .nu.: --SO2: 1333-1158
Step F:
(3R)-3-[3-(Morpholin-4-yl)propyl]-1,2,3,4-tetrahydroisoquinoline
[0166] To a solution of 1.50 g (3.62 mmol) of the compound obtained
in Step E in 20 mL of anhydrous methanol there are added 2.0 g
(82.3 mmol), in portions, of magnesium turnings. The batch is
stirred in the presence of ultrasound for 96 hours. The reaction
mixture is then filtered, the solid is washed several times with
methanol, and the filtrate is concentrated to dryness. After
purification by chromatography over silica gel
(dichloromethane/EtOH/NH.sub.4OH gradient), the expected product is
obtained in the form of an oil.
[0167] .sup.1H NMR (400 MHz, dmso-d6) .delta. ppm: 7.3 (d, 2H), 7.1
(t, 2H), 7.1 (d+t, 3H), 7 (d, 2H), 3.9 (s, 2H), 3.55 (t, 4H), 2.75
(m, 1H), 2.72/2.45 (dd, 2H), 2.35 (t, 4H), 2.25 (t, 2H), 1.6 (m,
2H), 1.45 (m, 2H)
[0168] IR (ATR) cm.sup.-1: .nu.: >NH2+/NH+: 3500-2300; .nu.:
>C--O--C<: 1115
[0169] High-Resolution Mass Spectrometry (ESI+/FIA/HR):
[0170] Empirical formula: C.sub.16H.sub.24N.sub.2O
[0171] [M+H].sup.+ calculated: 261.1961
[0172] [M+H].sup.+ measured: 261.1959
Preparation 4':
(3R)-3-(4-Morpholinylmethyl)-1,2,34-tetrahydroisoquinoline
[0173] The procedure is in accordance with the process of
Preparation 1', replacing the
(3S)-2-[(benzyloxy)carbonyl]-1,2,3,4-tetrahydro-3-isoquinolinecarboxylic
acid used in Step A by
(3R)-2-[(benzyloxy)carbonyl]-1,2,3,4-tetrahydro-3-isoquinolinecarboxylic
acid.
Preparation 1'':
4-{[tert-Butyl(dimethyl)silyl]oxy}-N-phenylaniline
[0174] To a solution of 12 g of 4-anilinophenol (64.7 mmol) in 200
mL of acetonitrile there are added, at ambient temperature, 6.7 g
of imidazole (97.05 mmol) and 11.7 g of
tert-butyl(chloro)dimethylsilane (77.64 mmol). The batch is stirred
at 70.degree. C. for 4 hours. The reaction mixture is then poured
into water and extracted with ether. The organic phase is then
dried over magnesium sulphate, then filtered and evaporated to
dryness. The crude product thereby obtained is then purified by
chromatography over silica gel (petroleum ether/dichloromethane
gradient). The title product is obtained in the form of a
powder.
[0175] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300K): 7.84 (s, 1H
NH); 7.17 (t, 2H aniline); 6.98 (d, 2H phenoxy); 6.94 (d, 2H
aniline); 6.76 (d, 2H phenoxy); 6.72 (t, 1H aniline); 0.95 (s, 9H
tert-butyl); 0.15 (s, 6H dimethyl)
[0176] IR: .nu.: >NH: 3403 cm.sup.-1; .nu.: >Ar: 1597
cm.sup.-1
Preparation 2'':
N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-indol-5-amine
[0177] The procedure is in accordance with the process of
Preparation 5'', replacing the 4-bromo-1-methyl-1H-pyrazole used in
Step B by 5-bromo-1-methyl-1H-indole.
Preparation 3'':
N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-pyrrolo[2,3-b]py-
ridin-5-amine
[0178] The procedure is in accordance with the process of
Preparation 5'', replacing the 4-bromo-1-methyl-1H-pyrazole used in
Step B by 5-bromo-1-methyl-1H-pyrrolo[2,3-b]pyridine (obtained in
accordance with a protocol from the literature: Heterocycles,
60(4), 865, 2003).
[0179] IR: .nu.: --NH--: 3278 cm.sup.-1; .nu.: aromatic --C.dbd.C--
moieties: 1605 cm.sup.-1
Preparation 4'':
N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)pyridin-4-amine
[0180] The procedure is in accordance with the process of
Preparation 5'', replacing the 4-bromo-1-methyl-1H-pyrazole used in
Step B by 4-bromopyridine.
[0181] IR: .nu. --NH--: 3200 and 2500 cm.sup.-1; .nu. --Si--O--:
902 cm.sup.-1; .nu. --Si--C--: 820 cm.sup.-1
Preparation 5'':
N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-pyrazol-4-amine
Step A: 4-{[tert-Butyl(dimethyl)silyl]oxy}aniline
[0182] The title compound is obtained starting from 4-aminophenol
in THF in the presence of imidazole and
tert-butyl(chloro)dimethylsilane in accordance with the protocol
described in the literature (S. Knaggs et al, Organic &
Biomolecular Chemistry, 3(21), 4002-4010; 2005).
[0183] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300K): 6.45-6.55
(dd, 4H, aromatic Hs); 4.60 (m, 2H, NH.sub.2-Ph); 0.90 (s, 9H, Si
(CH.sub.2).sub.2CH(CH.sub.3).sub.2); 0.10 (s, 6H, Si
(CH.sub.2).sub.2CH(CH.sub.3).sub.2)
[0184] IR: .nu.: --NH.sub.2.sup.+: 3300-3400 cm.sup.-1
Step B:
N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-1-methyl-pyrazol-4-amin-
e
[0185] To a solution of 30.8 g (0.137 mol) of the compound of Step
A in 525 mL of anhydrous toluene there are successively added 29.8
g of sodium tert-butylate (0.310 mol), 4.55 g of
Pd.sub.2(dba).sub.3 (also referred to as
tris(dibenzylideneacetone)dipalladium(0)) (4.96 mmol), 4.81 g of
2-di-tert-butylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl (9.91
mmol) and 12.8 mL of 4-bromo-1-methyl-1H-pyrazole (0.124 mol). The
batch is degassed under argon for 30 minutes and then refluxed for
3 hours. It is allowed to cool. The reaction mixture is
concentrated to dryness and then taken up in dichloromethane,
filtered over Celite and then concentrated to dryness again. The
residue is then purified by chromatography over silica gel
(gradient CH.sub.2Cl.sub.2/AcOEt) to provide the expected product
in the form of a solid.
[0186] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 300K): 7.55 (s, 1H,
pyrazole); 7.23 (s, 1H, pyrazole); 7.18 (broad s, 1H, NH.sub.2-Ph);
6.64 (m, 4H, aromatic Hs); 3.77 (s, 3H, CH.sub.3-pyrazole); 0.90
(s, 9H, Si (CH.sub.2).sub.2CH(CH.sub.3).sub.2); 0.12 (s, 6H, Si
(CH.sub.2).sub.2CH(CH.sub.3).sub.2)
[0187] IR: .nu. --NH.sup.+: 3275 cm.sup.-1; .nu. Ar and C.dbd.N:
1577 and 1502 cm.sup.-1; .nu. --Si--C--: 1236 cm.sup.-1; .nu.
--Si--O--: 898 cm.sup.-1; .nu. --Si--C--: 828, 774 cm.sup.-1
Preparation 6'':
N-{4-[(tert-Butyldimethylsilyl)oxy]phenyl}-1-trideuteriomethyl-1H-pyrazol-
-4-amine
Step A: 4-Bromo-1-trideuteriomethyl-1H-pyrazole
[0188] 4-Bromo-1H-pyrazole (9.05 g, 61.6 mmol) is added in portions
to a suspension of sodium hydride (60% in oil) (2.83 g, 70.8 mmol)
in tetrahydrofuran (90 mL) cooled in an ice bath. After having
taken away the ice bath, the solution is stirred at ambient
temperature for 0.5 hours. It is again cooled in an ice bath and
iodomethane-d.sub.3 (5.0 mL, 80.3 mmol) is added. The solution is
stirred at ambient temperature for 19 hours. The suspension is then
concentrated. The evaporation residue is triturated with tert-butyl
methyl ether (90 mL) and filtered. The filtrate is concentrated in
vacuo to obtain the expected compound in the form of an oil.
[0189] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm: 7.37 (s, 1H),
7.43 (s, 1H)
Step B:
N-{4-[(tert-Butyldimethylsilyl)oxy]phenyl}-1-trideuteriomethyl-1H--
pyrazol-4-amine
[0190] 4-Bromo-1-trideuteriomethyl-1H-pyrazole (9.6 g, 58.5 mmol),
4-[(tert-butyldimethyl-silyl)oxy]aniline (14.4 g, 64.6 mmol) and
toluene (150 mL) are added to a 500-ml three-necked flask. The
solution is degassed with nitrogen for 15 minutes, and then sodium
tert-butylate (11.4 g, 0.12 mol),
2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (0.77 g,
1.81 mmol) and tris(dibenzylideneacetone)dipalladium(0) (1.64 g,
1.79 mmol) are successively added. The suspension is heated at
85.degree. C. for 1.5 hours. The reaction mixture is then cooled to
ambient temperature and water (270 mL) is added. The mixture is
stirred for 30 minutes. Celite (30 g) is then added and the
suspension is filtered on a bed of Celite. The phases of the
filtrate are separated and the aqueous phase is extracted with
ethyl acetate (3.times.200 mL). The combined organic phases are
dried over sodium sulphate and filtered. The product is purified by
chromatography over silica gel (ethyl acetate/heptane gradient).
The product obtained is recrystallized from heptane (80 mL) to
obtain the expected compound.
[0191] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm: 0.16 (s, 6H),
0.97 (s, 9H), 4.92 (s, 1H), 6.61-6.73 (m, 4H), 7.25 (s, 1H), 7.36
(s, 1H)
[0192] .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. ppm: -4.37,
18.28, 25.86, 38.67 (sept., .sup.1J.sub.C-D=21.0 Hz), 115.12,
120.73, 123.76, 126.52, 134.74, 141.07, 148.43
[0193] MS (ESI): [M+H].sup.+ 307.08
Preparation 7'':
4-({4-[(tert-Butyldimethylsilyl)oxy]phenyl}amino)-1,5-dimethyl-1H-pyrrole-
-2-carbonitrile
Step A: 4-Bromo-1,5-dimethyl-1H-pyrrole-2-carbonitrile
[0194] A solution of bromine (6.58 mL, 0.13 mol) in acetic acid (60
mL) is added dropwise, with the aid of a dropping funnel, to a
solution of 1,5-dimethyl-1H-pyrrole-2-carbonitrile (15.0 g, 0.12
mol) in acetic acid (300 mL). The batch is stirred at ambient
temperature for 24 hours. The reaction mixture is then poured into
a beaker containing 300 mL of water. The solid formed is filtered
off and rinsed with water. It is then dissolved in dichloromethane
(300 mL) and the organic phase is washed with brine, dried over
sodium sulphate, filtered and concentrated in vacuo to yield the
expected product in the form of a solid.
[0195] .sup.1H NMR (CDCl.sub.3) .delta. ppm: 2.25 (s, 3H), 3.67 (s,
3H), 6.74 (s, 1H)
Step B:
4-({4-[(tert-Butyldimethylsilyl)oxy]phenyl}amino)-1,5-dimethyl-1H--
pyrrole-2-carbonitrile
[0196] A solution of the compound of the above Step (1.5 g, 7.53
mmol), 4-[(tert-butyldimethylsilyl)oxy]aniline (2.02 g, 9.04 mmol),
sodium tert-butylate (1.45 g, 15.06 mmol) and
2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (0.13 g,
0.30 mmol) in toluene (20 mL) is purged with nitrogen.
Tris(dibenzylideneacetone)-dipalladium(0) (0.28 g, 0.30 mmol) is
added, and then the reaction mixture is heated at 90.degree. C.
until the reaction is complete (monitored by TLC). Heating is
stopped and the mixture is allowed to return to ambient
temperature. Water (75 mL) is added and the mixture is extracted
with ethyl acetate (3.times.75 mL). The combined organic phases are
washed with brine and then concentrated. The crude product is
purified by flash chromatography over silica gel (ethyl
acetate/heptane gradient). The product thereby obtained is
dissolved in heptane in the warm state and is allowed to
precipitate, with stirring, at ambient temperature, and then at
0.degree. C. The solid is filtered off and the operation is
repeated on the filtrate to yield the expected compound in the form
of a solid.
[0197] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm: 0.15 (s, 6H),
0.97 (s, 9H), 2.13 (s, 3H), 3.66 (s, 3H), 4.68 (br. s, 1H), 6.49
(d, J=8.5 Hz, 2H), 6.64 (s, 1H), 6.66 (d, J=8.7 Hz, 2H)
[0198] .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. ppm: 4.34, 9.72,
18.30, 25.88, 32.94, 101.27, 114.37, 114.70, 116.41, 120.73,
124.52, 131.23, 141.54, 148.27
[0199] MS (ESI+): [M+H].sup.+ measured: 342.3
Preparation 8'':
4-[(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)amino]-1-methyl-1H-pyrrole-2-
-carbonitrile
Step A: 1-Methyl-1H-pyrrole-2-carbonitrile
[0200] N,N-Dimethylformamide (3 mL) and
1,4-diazabicyclo[2.2.2]octane (0.49 g, 4.3 mmol) are added to a
solution of pyrrole-2-carbonitrile (4 g, 43.4 mmol) in dimethyl
carbonate (56 mL). The solution is stirred at 90.degree. C. for 15
hours, and is then heated at 110.degree. C. for 8 hours. The
mixture is cooled to ambient temperature, and then ethyl acetate
(80 mL) is added. The phases are separated and the organic phase is
washed with water (2.times.80 mL) and 1M aqueous hydrochloric acid
solution (1.times.80 mL). The combined aqueous phases are extracted
again with ethyl acetate (1.times.80 mL). The combined organic
phases are washed with brine (1.times.80 mL), dried over magnesium
sulphate, filtered and concentrated in vacuo to obtain the expected
product in the form of a liquid.
[0201] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm: 3.78 (m, 2H),
6.12-6.18 (m, 1H), 6.74-6.82 (m, 1H)
Step B: 4-Bromo-1-methyl-1H-pyrrole-2-carbonitrile
[0202] N-Bromosuccinimide (6.2 g, 34.9 mmol) is added to a solution
of 1-methyl-1H-pyrrole-2-carbonitrile (3.7 g, 34.9 mmol) in
N,N-dimethylformamide (150 mL). The solution is stirred for 15
hours at ambient temperature. Another amount of N-bromosuccinimide
(2.0 g, 11 mmol) is added and the mixture is stirred for 3 hours.
The product is purified by chromatography over silica gel (ethyl
acetate/heptane gradient) to obtain the expected product in the
form of a solid.
[0203] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm: 3.77 (s, 3H),
6.75 (d, J=1.7 Hz, 1H), 6.80 (d, J=1.7 Hz, 1H)
Step C:
4-[(tert-Butyldimethylsilyl)oxy]phenyl}amino)-1-methyl-1H-pyrrole--
2-carbonitrile
[0204] Nitrogen is bubbled through a solution of
4-bromo-1-methyl-1H-pyrrole-2-carbonitrile (2.82 g, 15.2 mmol) and
4-[(tert-butyldimethylsilyl)oxy]aniline (4.08 g, 18.3 mmol) in
toluene (55 mL) for 5 minutes. Sodium tert-butylate (2.92 g, 30.4
mmol), tris(dibenzylideneacetone)dipalladium(0) (556 mg, 0.6 mmol)
and 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (255 mg,
0.6 mmol) are then added to the reaction mixture. The mixture is
stirred for 1 hour at 80.degree. C. under nitrogen. The suspension
is then cooled to ambient temperature and filtered over Celite. The
Celite cake is then rinsed with ethyl acetate. The filtrate is
washed with water and then with brine. The organic phase is dried
over magnesium sulphate, filtered and concentrated in vacuo. The
product is purified twice by chromatography over silica gel
(AcOEt/heptane gradient), and then by trituration in heptane to
obtain the expected product in the form of a solid.
[0205] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm: 0.16 (s, 6H),
0.97 (s, 9H), 3.73 (s, 3H), 6.57 (d, J=1.9 Hz, 1H), 6.64-6.66 (m,
1H), 6.70 (s, 4H); NMR
[0206] .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. ppm: -4.48,
18.17, 25.72, 35.46, 103.01, 113.56, 113.69, 115.92, 119.55,
120.67, 129.04, 139.94, 148.85
[0207] MS (ESI+): [M+H].sup.+ 328.25
[0208] The amines NHR.sub.3R.sub.4 wherein R.sub.3 and R.sub.4,
each independently of the other, represent an aryl or heteroaryl
group are obtained in accordance with processes described in the
literature (Surry D. S. et al., Chemical Science, 2011, 2, 27-50,
Charles M. D. et al., Organic Letters, 2005, 7, 3965-3968). The
reaction protecting the hydroxy function of the 4-anilinophenol
described in Preparation 1'' can be applied to various secondary
amines NHR.sub.3R.sub.4 (as defined hereinbefore) having one or
more hydroxy functions, when they are available commercially.
Alternatively, the secondary amines having at least one hydroxy
substituent may be synthesised directly in a protected form, i.e.
starting from reagents whose hydroxy function has been protected
beforehand. Among the protecting groups,
tert-butyl(dimethyl)silyloxy and benzyloxy are especially
preferred.
[0209] Among the amines NHR.sub.3R.sub.4 having a hydroxy
substituent that are used for synthesising the compounds of the
invention there may be mentioned: 4-(4-toluidino)phenol,
4-(4-chloroanilino)phenol, 4-(3-fluoro-4-methylanilino)phenol,
4-[4-(trifluoromethoxy)anilino]phenol, 4-[4-hydroxyanilino]phenol,
{4-[(1-methyl-1H-indol-6-yl)amino]phenyl}methanol,
4-(2,3-dihydro-1H-indol-6-ylamino)phenol,
4-[(1-methyl-2,3-dihydro-1H-indol-6-yl)amino]phenol,
4-[(1-methyl-1H-indol-6-yl)amino]phenol,
4-[(1-methyl-1H-indol-6-yl)amino]cyclohexanol,
4-[(1-methyl-1,2,3,4-tetrahydro-6-quinolinyl)amino]phenol,
4-[(4-methyl-3,4-dihydro-2H-1,4-benzoxazin-7-yl)amino]phenol,
4-[4-(diethylamino)anilino]phenol,
4-(2,3-dihydro-1H-inden-5-ylamino)phenol,
4-[(1-methyl-1H-indazol-5-yl)amino]phenol,
4-[(1'-methyl-1',2'-dihydrospiro[cyclopropane-1,3'-indol]-5'-yl)amino]phe-
nol, 4-[(1,3,3-trimethyl-2,3-dihydro-1H-indol-5-yl) amino] phenol,
4-[4-methoxy-3-(trifluoromethyl)anilino]phenol,
4-[4-(methylsulphanyl)-3-(trifluoromethyl)anilino]phenol,
2-fluoro-4-[(1-methyl-1H-indol-5-yl)amino]phenol,
4-[(1-ethyl-1H-indol-5-yl)amino]phenol,
4-[(1-ethyl-2,3-dihydro-1H-indol-5-yl)amino]phenol,
4-[(1-isopropyl-2,3-dihydro-1H-indol-5-yl)amino]phenol,
4-(butylamino)phenol, 3-[(1-methyl-1H-indol-5-yl)amino]-1-propanol,
4-[(1-methyl-1H-indol-5-yl)amino]-1-butanol,
4-[(3-fluoro-4-methylphenyl)amino]phenol,
4-[(3-chloro-4-methylphenyl)amino]phenol,
4-[(4-fluorophenyl)amino]phenol,
4-[(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenol,
4-[(4-fluorophenyl)amino]phenol, 4-[(2-fluorophenyl)amino]phenol,
4-[(3-fluorophenyl)amino]phenol,
4-[(2,4-difluorophenyl)amino]phenol,
4-[(3,4-difluorophenyl)amino]phenol,
3-[(4-hydroxyphenyl)amino]benzonitrile,
4-[(3-methoxyphenyl)amino]phenol,
4-[(3,5-difluorophenyl)amino]phenol,
4-[(3-methylphenyl)amino]phenol,
4-[(4-hydroxyphenyl)amino]benzonitrile,
4-[(3-chlorophenyl)amino]phenol, 4-(pyrimidin-2-ylamino)phenol,
4-[(cyclobutylmethyl)amino]phenol,
2-[(4-hydroxyphenyl)amino]benzonitrile,
4-{[(1-methyl-1H-pyrazol-4-yl)methyl]amino}phenol,
4-[(cyclopropylmethyl)amino]phenol,
4-{[(1-methyl-1H-pyrazol-3-yl)methyl]amino}phenol,
4-(but-2-yn-1-ylamino)phenol, 4-(pyrazin-2-ylamino)phenol,
4-(pyridin-2-ylamino)phenol, 4-(pyridazin-3-ylamino)phenol,
4-(pyrimidin-5-ylamino)phenol, 4-(pyridin-3-ylamino)phenol,
4-[(3,5-difluoro-4-methoxyphenyl)amino]phenol,
4-(pyridin-4-ylamino)phenol,
4-[(3-fluoro-4-methoxyphenyl)amino]phenol,
2-(phenylamino)pyrimidin-5-ol,
5-[(4-hydroxyphenyl)amino]-2-methoxybenzonitrile and
4-{[3-(trifluoromethyl)phenyl]amino}phenol.
[0210] The hydroxy function(s) of the secondary amines listed above
is (are) protected beforehand by a suitable protecting group prior
to any coupling to an acid derivative of the compound of formula
(VII) as defined in the preceding general process.
EXAMPLE 1
4-[{[3-(6-{[(3S)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]-
carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}-
(phenyl)amino]phenyl disodium phosphate
Step A: Methyl
3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)-carb-
onyl]-1,3-benzodioxol-5-yl}-5,6,7,8-tetrahydro-1-indolizine-carboxylate
[0211] To a solution of 2 g of the compound of Preparation 1 (5.83
mmol) in 20 mL of dichloromethane there are added, at ambient
temperature, 5.5 mL of N,N,N-triethylamine (6.96 mmol), 2.12 g of
the compound of Preparation 1' (6.96 mmol), and then 0.94 g of
hydroxybenzotriazole (HOBT) and 1.34 g of
1-ethyl-3-(3'-dimethylaminopropyl)-carbodiimide (EDC) (6.96 mmol).
The reaction mixture is then stirred at ambient temperature
overnight and then it is poured into saturated aqueous ammonium
chloride solution and extracted with ethyl acetate. The organic
phase is then dried over magnesium sulphate and then filtered and
evaporated to dryness. The crude product thereby obtained is then
purified by chromatography over silica gel (heptane/AcOEt
gradient). The title product is obtained in the form of an oil.
[0212] .sup.1H NMR: .delta. (500 MHz; dmso-d6; 300.degree. K):
7.2-6.9 (m, 4H, aromatic Hs); 7.04-7.03-7.00 (m, 1H, aromatic H);
6.85 (m, 1H, aromatic H); 6.35-6.26-6.06 (m, 1H, H
tetrahydroindolizine); 6.15-6.12 (m, 2H, H methylenedioxy);
5.06-4.84 (m, 1H, H dihydroisoquinoline); 4.86-4.17 (m, 2H, H
dihydroisoquinoline); 3.65-3.6-3.55 (m, 3H, H methyl ester);
3.43-4.26 (m, 2H, H tetrahydroindolizine); 3.58-3.5 (m, 4H, H
morpholine); 2.37-3.05 (m, 4H, 2H dihydroisoquinoline, 2H
tetrahydroindolizine); 1.68-2.56 (m, 4H, H morpholine); 1.4-2.0 (m,
4H, H tetrahydroindolizine)
[0213] IR: .nu.: >C.dbd.O 1695 cm.sup.-1 ester; .nu.:
>C.dbd.O 1625 cm.sup.-1 amide; .nu.: >C--O--C<
1214-1176-1115 cm.sup.-1; >CH--Ar 772-744 cm.sup.-1
Step B: Lithium
3-[6-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1-
,3-benzodioxol-5-yl]-5,6,7,8-tetrahydro-1-indolizine-carboxylate
[0214] To a solution of 4.6 g of the compound of Step A (8.26 mmol)
in 24 mL of dioxane there is added a solution of lithium hydroxide
(675 mg, 16.1 mmol). The batch is placed in a microwave oven at 140
W, 100.degree. C. for a period of 2 hours 30 minutes. The reaction
mixture is then filtered and evaporated. The solid thereby obtained
is dried at 40.degree. C. in an oven in the presence of
P.sub.2O.sub.5.
[0215] .sup.1H NMR: .delta. (400 MHz; dmso-d6; 353.degree. K):
6.7-7.15 (unresolved peak, 6H, aromatic Hs); 6.21 (s, 1H, aromatic
H); 6.03 (s, 2H, H methylenedioxy); 4.0-5.0 (unresolved peak, 3H
dihydroisoquinoline); 3.4-3.6 (unresolved peak, 3H
tetrahydroindolizine, 3H morpholine); 2.5-3.1 (unresolved peak, 4H,
2H tetrahydroindolizine, 2H morpholine); 1.5-2.4 (unresolved peak,
10H morpholine)
[0216] IR: .nu.: >C.dbd.O broad 1567 cm.sup.-1 acetate; .nu.:
1236 cm.sup.-1
Step C:
N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3-{6-[((3S)-3-(4-morp-
holinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-
-yl}-N-phenyl-5,6,7,8-tetrahydro-1-indolizine carboxamide
[0217] To a solution of 2.6 g of the compound of Step B (4.73 mmol)
in 47 mL of dichloromethane there are added, dropwise, 1.2 mL of
oxalyl chloride (14.2 mmol) at 0.degree. C. The reaction mixture is
stirred at ambient temperature for 11 hours and then co-evaporated
several times with dichloromethane. The product thereby obtained is
suspended in 37 mL of dichloromethane, and is then added to a
solution of 2.1 g of the compound obtained in Preparation 1'' (7.1
mmol) in 10 mL of dichloromethane in the presence of 0.6 mL of
pyridine (7.1 mmol). The batch is stirred at ambient temperature
overnight.
[0218] The reaction mixture is concentrated and purified by
chromatography over silica gel (dichloromethane/methanol gradient).
The title product is obtained in the form of a foam.
[0219] .sup.1H NMR: .delta. (500MHz; dmso-d6; 300.degree. K):
6.9-7.3 (9H, aromatic Hs); 6.88 (2H, aromatic Hs); 6.72-6.87 (2H,
aromatic Hs); 6.64 (2H, aromatic Hs); 6.13 (2H methylenedioxy);
5.05-4.74 (1H dihydroisoquinoline); 4.25-4.13 (2H
dihydroisoquinoline); 3.44-3.7 (4H morpholine); 3.62-3.52 (2H
tetrahydroindolizine); 3.0-2.6 (4H, 2H tetrahydroindolizine, 2H
dihydroisoquinoline); 2.54-1.94 (6H morpholine); 1.91-1.53 (4H
tetrahydroindolizine); 0.92 (9H tert-butyl); 0.17 (6H dimethyl)
[0220] IR: .nu.: >C.dbd.O: 1632 cm.sup.-1; .nu.:
>C--O--C<: 1237 cm.sup.-1; .nu.: --Si--O--C--: 1035
cm.sup.-1; --Si--C--: 910 cm.sup.-1; >CH--Ar: 806 cm.sup.-1
Step D:
N-(4-Hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydr-
o-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tet-
rahydro-1-indolizine carboxamide hydrochloride
[0221] To a solution of 1.9 g of the compound obtained in Step C
(2.3 mmol) in 4 mL of methanol there is added 0.646 g (11.5 mmol)
of potassium hydroxide dissolved in 8 mL of methanol. The batch is
stirred at ambient temperature for 30 minutes. The reaction mixture
is then diluted with dichloromethane and washed successively with
1M HCl solution, saturated aqueous NaHCO.sub.3 solution and then
brine until a neutral pH is reached. The organic phase is then
dried over magnesium sulphate, filtered and evaporated. The crude
product thereby obtained is purified by chromatography over silica
gel (dichloromethane/methanol gradient). The solid is then
dissolved in dichloromethane, and 2 mL of 1M ethereal HCl are
added. The batch is stirred for 1 hour and then evaporated to
dryness. The hydrochloride thereby obtained is dissolved in a
mixture of water/acetonitrile until dissolution is complete, and is
then lyophilised.
[0222] Elemental Microanalysis: (%, Theoretical:Measured)
[0223] % C=69.11:68.95; % H=5.8:5.46; % N=7.5:7.51; %
Cl-=4.74:4.48
[0224] Optical rotation: (.alpha.).sub.D.sup.20+50.8.degree. (c=9
mg/mL, MeOH)
Step E:
4-[{[3-(6-{[(3S)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2-
(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-ylic-
arbonyl}(phenyl)amino]phenyl dibenzyl phosphate
[0225] To a suspension of 82 mg of sodium hydride (2.06 mmol) in 10
mL of anhydrous THF there are added, in portions and at 0.degree.
C., 700 mg of the compound of Step D. After stirring for 30 minutes
at 0.degree. C. and for 30 minutes at ambient temperature,
tetrabenzyl pyrophosphate is added at 0.degree. C. and the reaction
mixture is stirred overnight at ambient temperature. After
evaporating off the solvent, the crude reaction product is diluted
with dichloromethane (30 mL), washed with saturated aqueous
NaHCO.sub.3 solution and then with brine. The organic phase is then
dried over MgSO.sub.4, filtered, concentrated to dryness and
purified by chromatography over silica gel (gradient
CH.sub.2Cl.sub.2/MeOH). The title product is then obtained in the
form of a solid.
[0226] .sup.1H NMR: .delta. (500 MHz; DMSO-d6; 300K): 7.34 (m, 10H,
phenyl); 7.30-6.71 (m, 15H, aryl); 6.06 (s, 1H, methylenedioxy);
5.30-4.97 (m, 1H, pyrrole); 5.11 (m, 4H, benzyl): 5.03-3.64 (m, 1H,
tertiary C THIQ); 4.91-4.09 (m, 2H, secondary C THIQ); 3.99-3.48
(m, 2H, secondary C THIQ); 3.54-3.44 (m, 4H, morpholine); 2.89-2.65
(m, 3H, secondary C THIQ); 2.51-1.87 (m, 4H, secondary C THID);
2.36-1.85 (m, 2H, secondary C THIQ); 1.91-1.45 (m, 4H, secondary C
THID)
Step F:
4-[{[3-(6-{[(3S)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2-
(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]c-
arbonyl}(phenyl)amino]phenyl disodium phosphate
[0227] 50 mg of Pd(OH).sub.2 are added to a solution of the product
obtained in Step E (505 mg; 0.52 mmol) in methanol (10 mL), and
then the reaction mixture is placed under a hydrogen atmosphere (1
bar) for 5 hours. After filtering off the catalyst and
concentrating to dryness, the crude reaction product is dissolved
in methanol (5 mL) and treated with 0.95 mL of 1M sodium hydroxide
solution. The solvents are then evaporated off and the crude
reaction product is purified by chromatography over an OASIS.RTM.
phase (acetonitrile/H.sub.2O gradient) to obtain a white solid.
[0228] Elemental Microanalysis:
TABLE-US-00001 % C % H % N % Na Calculated 61.87 4.95 6.71 5.51
Found 61.45 4.46 6.61 5.38
[0229] IR: .nu.: --C.dbd.O: 1628 cm.sup.-1; .nu.: C--O--C: 1234
cm.sup.-1; .nu.: P.dbd.O: 115 cm.sup.-1; .nu.: P--O: 985 cm.sup.-1;
.nu.: CH--Ar: 876 cm.sup.-1
[0230] High-Resolution Mass Spectrometry (ESI+):
[0231] Empirical formula:
C.sub.43H.sub.41N.sub.4Na.sub.2O.sub.9P
[0232] [M+H]+ calculated: 835.2479
[0233] [M+H]+ measured: 835.2467
EXAMPLE 2
4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-be-
nzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]ph-
enyl disodium phosphate
Step A:
N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(-
1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizi-
ne-1-carboxamide
[0234] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing the product of
Preparation 1' in Step A by that of Preparation 2', it being
understood that the product thereby obtained is not subjected to a
step of conversion into salt form in the presence of HCl in
ether.
[0235] Elemental Microanalysis: (%, Theoretical:Measured)
[0236] % C=74.86:74.88; % H=5.64:5.31; % N=6.72:6.78
Step B:
4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl-
}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)-
amino]phenyl disodium phosphate
[0237] The procedure is in accordance with a protocol analogous to
that described in Steps E and F of Example 1.
[0238] High-Resolution Mass Spectrometry (ESI+):
[0239] Empirical formula: C.sub.39H.sub.36N.sub.3O.sub.8P
[0240] [M+H].sup.+ calculated: 706.2313
[0241] [M+H]+ measured: 706.2324
EXAMPLE 3
4-[(1-Methyl-1H-indol-5-yl){[3-(2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihy-
droisoquinolin-2(1H)-yl]carbonyl}phenyl)-5,6,7,8-tetrahydroindolizin-1-yl]-
carbonyl}amino]phenyl disodium phosphate
Step A:
3-{5-Chloro-2-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-iso-
quinolinyl)carbonyl]phenyl}-N-(4-hydroxyphenyl)-N-(1-methyl-1H-indol-5-yl)-
-5,6,7,8-tetrahydro-1-indolizine carboxamide hydrochloride
[0242] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 2 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 2''.
[0243] Elemental Microanalysis:
TABLE-US-00002 % C % H % N % Cl.sup.- Calculated 68.04 5.72 8.82
4.91 Found 67.84 5.46 8.64 5.21
[0244] Optical rotation: (.alpha.).sub.D.sup.20+55.9.degree. (c=7
mg/mL, MeOH)
Step B:
4-[(1-Methyl-1H-indol-5-yl){[3-(2-{[(3S)-3-(morpholin-4-ylmethyl)--
3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-5,6,7,8-tetrahydroindoliz-
in-1-yl]carbonyl}amino]phenyl disodium phosphate
[0245] The procedure is in accordance with a protocol analogous to
that described in Steps E and F of Example 1.
[0246] High-Resolution Mass Spectrometry (ESI+):
[0247] Empirical formula:
C.sub.45H.sub.44N.sub.5Na.sub.2O.sub.7P
[0248] [M-2Na+3H].sup.+ calculated: 800.3208
[0249] [M-2Na+3H]+ measured: 800.3211
EXAMPLE 4
4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-be-
nzodioxol-5-yl)indolizin-1-yl]carbonyl}(1-methyl-1H-pyrrolo[2,3-b]pyridin--
5-yl)amino]phenyl disodium phosphate
Step A:
N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(-
1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridi-
n-5-yl)-indolizine-1-carboxamide hydrochloride
[0250] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compounds of Preparations 1 and 1' used in Step A by the
compounds of Preparations 3 and 2' and, on the other hand, the
compound of Preparation 1'' used in Step C by that of Preparation
3''.
[0251] Elemental Microanalysis: (%, Theoretical:Measured)
[0252] % C=69.14:70.09; % H=4.81:4.55; % N=9.83:10.09; %
Cl-=4.98:3.26
Step B:
4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl-
}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(1-methyl-1H-pyrrolo[2,3-b]-
pyridin-5-yl)amino]phenyl diethyl phosphate
[0253] To a suspension of the compound obtained in Step A (1.5
mmol) in 10 mL of anhydrous CH.sub.2Cl.sub.2 there are added
triethylamine (0.42 mL; 3 mmol), and then diethyl cyanophosphonate
(0.24 mL; 1.65 mmol) dropwise at ambient temperature. After
stirring overnight at ambient temperature, the reaction mixture is
diluted with CH.sub.2Cl.sub.2, washed with saturated aqueous
NaHCO.sub.3 solution and then with brine. The organic phase is then
dried over MgSO.sub.4, filtered, concentrated to dryness and
purified by chromatography over silica gel (CH.sub.2Cl.sub.2/MeOH
gradient). The title product is then obtained in the form of a
solid.
Step C:
4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl-
}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(1-methyl-1H-pyrrolo[2,3-b]-
pyridin-5-yl)amino]phenyl disodium phosphate
[0254] 0.4 mL of trimethylsilyl bromide (3 mmol) is added dropwise
at ambient temperature to a solution of the product obtained in
Step B (0.78 mmol) in CH.sub.2Cl.sub.2 (12 mL). The reaction
mixture is stirred for 5 hours, and then a solution of
Na.sub.2CO.sub.3 (580 mg) in water (4 mL) is slowly added at
0.degree. C. After stirring for 30 minutes, the reaction mixture is
concentrated to dryness, diluted with anhydrous methanol (25 mL)
and microfiltered. The filtrate is dried and purified by
chromatography over an OASIS.RTM. phase (acetonitrile/H.sub.2O
gradient).
[0255] High-Resolution Mass Spectrometry (ESI+):
[0256] Empirical formula:
C.sub.45H.sub.44N.sub.5Na.sub.2O.sub.7P
[0257] [M-2Na+3H].sup.+ calculated: 800.3211
[0258] [M-2Na+3H]+ measured: 800.3201
EXAMPLE 5
4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-be-
nzodioxol-5-yl)indolizin-1-yl]carbonyl}(pyridin-4-yl)amino]phenyl
disodium phosphate
Step A:
N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(-
1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(pyridin-4-yl)indolizine-1-carbox-
amide hydrochloride
[0259] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compounds of Preparations 1 and 1' used in Step A by the
compounds of Preparations 3 and 2' and, on the other hand, the
compound of Preparation 1'' used in Step C by that of Preparation
4''.
[0260] Elemental Microanalysis: (%, Theoretical:Measured)
[0261] % C=69.24:69.12; % H=4.74:4.23; % N=8.5:8.45; %
Cl-=5.38:5.2
Step B:
4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl-
}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(pyridin-4-yl)amino]phenyl
diethyl phosphate
[0262] To a suspension of 950 mg of the compound obtained in Step A
(1.5 mmol) in 10 mL of anhydrous CH.sub.2Cl.sub.2 there are added
triethylamine (0.42 mL; 3 mmol), and then diethyl cyanophosphonate
(0.24 mL; 1.65 mmol) dropwise at ambient temperature. After
stirring overnight at ambient temperature, the reaction mixture is
diluted with CH.sub.2Cl.sub.2, washed with saturated aqueous
NaHCO.sub.3 solution and then with brine. The organic phase is then
dried over MgSO.sub.4, filtered, concentrated to dryness and
purified by chromatography over silica gel (CH.sub.2Cl.sub.2/MeOH
gradient). The title product is then obtained in the form of a
solid.
[0263] .sup.1H NMR: .delta. (500 MHz; DMSO-d6; 300K): 8.5-8. 0 (m,
5H); 7.2-7.1 (m, 1H); 6.85-6.65 (m, 1H); 7.3-6.8 (m, 10H);
6.25-6.10 (bs, 1H); 6.2 (bs, 2H); 5.1-3.7 (6d, 2H); 4.7-3.8 (m,1H);
4.15 (m, 4H); 3.0-1.7 (m,2H); 1.25 (m, 6H); 0.85-0.24 (m,3H)
Step C:
4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl-
}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(pyridin-4-yl)amino]phenyl
disodium phosphate
[0264] 0.4 mL of trimethylsilyl bromide (3 mmol) is added dropwise
at ambient temperature to a solution of the product obtained in
Step B (591 mg; 0.78 mmol) in CH.sub.2Cl.sub.2 (12 mL). The
reaction mixture is stirred for 5 hours, and then a solution of
Na.sub.2CO.sub.3 (580 mg) in water (4 mL) is slowly added at
0.degree. C. After stirring for 30 minutes, the reaction mixture is
concentrated to dryness, diluted with anhydrous methanol (25 mL)
and microfiltered. The filtrate is dried and purified by
chromatography over an OASIS.RTM. phase (acetonitrile/H.sub.2O
gradient).
[0265] .sup.1H NMR: .delta. (500 MHz; D.sub.2O; 300K): 8.23-7.98
(m, 2H, pyridyl); 7.01-6.97 (m, 2H, pyridyl); 7.88-7.80 (m, 1H,
indolizine); 7.18-6.57 (m, 13H, aromatic Hs
THIQ+aryl+indolizine+phenol); 6.17-6.15 (m, 1H, indolizine): 5.96
(m, 2H, methylenedioxy); 4.61-3.76 (m, 1H, ternary C THIQ); 4.16
(m, 2H, secondary C THIQ); 2.86-2.31 (m, 2H, secondary C THIQ);
0.94-0.76 (m, 3H, primary C THIQ)
[0266] IR: .nu.: --C.dbd.O: 1620 cm.sup.-1; .nu.: C--O--C: 1218
cm.sup.-1; .nu.: P.dbd.O: 1107 cm.sup.-1 .nu.: P--O: 981 cm.sup.-1
.nu.: CH--Ar: 881-741 cm.sup.-1
[0267] High-Resolution Mass Spectrometry (ESI+):
[0268] Empirical formula:
C.sub.38H.sub.29N.sub.4Na.sub.2O.sub.8P
[0269] [M-2Na+3H].sup.+ calculated: 703.1952
[0270] [M-2Na+3H]+ measured: 703.1951
EXAMPLE 6
4-[{[3-(6-{[(3S)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]-
carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}-
(phenyl) amino]phenyl dibenzyl phosphate
[0271] The procedure is in accordance with the protocol described
in Steps A-E of Example 1.
[0272] .sup.1H NMR: .delta. (500 MHz; DMSO-d6; 300K): 7.34 (m, 10H,
phenyl); 7.30-6.71 (m, 15H, aryl); 6.06 (s, 1H, methylenedioxy);
5.30-4.97 (m, 1H, pyrrole); 5.11 (m, 4H, benzyl): 5.03-3.64 (m, 1H,
tertiary C THIQ); 4.91-4.09 (m, 2H, secondary C THIQ); 3.99-3.48
(m, 2H, secondary C THIQ); 3.54-3.44 (m, 4H, morpholine); 2.89-2.65
(m, 3H, secondary C THIQ); 2.51-1.87 (m, 4H, secondary C THID);
2.36-1.85 (m, 2H, secondary C THIQ); 1.91-1.45 (m, 4H, secondary C
THID)
EXAMPLE 7
Diethyl
4-[{[3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl-
}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(pyridin-4-yl)amino]phenyl
phosphate
[0273] The procedure is in accordance with the protocol described
in Steps A and B of Example 5.
[0274] .sup.1H NMR: .delta. (500 MHz; DMSO-d6; 300K): 8.5-8. 0 (m,
5H); 7.2-7.1 (m, 1H); 6.85-6.65 (m, 1H); 7.3-6.8 (m, 10H);
6.25-6.10 (bs, 1H); 6.2 (bs, 2H); 5.1-3.7 (6d, 2H); 4.7-3.8 (m,
1H); 4.15 (m, 4H); 3.0-1.7 (m,2H); 1.25 (m, 6H); 0.85-0.24 (m,
3H)
EXAMPLE 8
4-[{[3-(6-{[(3S)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]-
carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}-
(phenyl)amino]phenyl dihydrogen phosphate hydrochloride
[0275] 100 mg of Pd(OH).sub.2 are added to a solution of the
product obtained in Step E of Example 1 (500 mg; 0.51 mmol) in
methanol (10 mL), and then the reaction mixture is placed under a
hydrogen atmosphere (1 bar) for 5 hours. After filtering off the
catalyst and concentrating to dryness, the crude reaction product
is immediately purified by chromatography over a C18 phase
(acetonitrile/H.sub.2O+0.2% HCl gradient) to obtain a solid.
[0276] High-Resolution Mass Spectrometry (ESI+):
[0277] Empirical formula: C.sub.43H.sub.43N.sub.4O.sub.9P
[0278] [M+H].sup.+ calculated: 791.2846
[0279] [M+H].sup.+ measured: 791.2852
EXAMPLE 9
4-[{[5-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbony-
l}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-yl)amino]phenyl
disodium phosphate
Step A:
5-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carb-
onyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-N-(pyridin-4-yl)-1H-pyrrole--
3-carboxamide hydrochloride
[0280] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compounds of Preparations 1 and 1' used in Step A by the
compounds of Preparations 4 and 2' and, on the other hand, the
compound of Preparation 1'' used in Step C by that of Preparation
4''.
[0281] Elemental Microanalysis: (%, Theoretical:Measured)
[0282] % C=66.99:66.88; % H=5.14:5.28; % N=8.93:8.87; %
Cl-=5.65:4.98
[0283] High-Resolution Mass Spectrometry (ESI+):
[0284] Empirical formula: C.sub.35H.sub.32ClN.sub.4O.sub.3
[0285] [M+H].sup.+ calculated: 591.2157
[0286] [M+H].sup.+ measured: 591.2178
Step B:
4-[{[5-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl-
]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-yl)amino-
]phenyl disodium phosphate
[0287] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 4.
EXAMPLE 10
4-[{[1,2-Dimethyl-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquino-
lin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbonyl}(1-me-
thyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyl disodium
phosphate
Step A:
N-(4-Hydroxyphenyl)-1,2-dimethyl-N-(1-methyl-1H-pyrrolo[2,3-b]pyri-
din-5-yl)-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H-
)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrole-3-carboxamide
hydrochloride
[0288] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 5 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 3''.
[0289] Elemental Microanalysis: (%, Measured(Theoretical))
[0290] % C=66.41(66.62); % H=5.08(5.59); % N=10.85(10.84); %
Cl-=4.68(4.57)
Step B:
4-[{[1,2-Dimethyl-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydro-
isoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbon-
yl}(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyl disodium
phosphate
[0291] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 4.
EXAMPLE 11
4-[{[1,2-Dimethyl-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquino-
lin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbonyl}(1-me-
thyl-1H-pyrazol-4-yl)amino]phenyl disodium phosphate
Step A:
N-(4-Hydroxyphenyl)-1,2-dimethyl-N-(1-methyl-1H-pyrazol-4-yl)-5-(6-
-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-
-1,3-benzodioxol-5-yl)-1H-pyrrole-3-carboxamide hydrochloride
[0292] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 5 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 5''.
[0293] Elemental Microanalysis: (%, Measured(Theoretical))
[0294] % C=64.25(64.59); % H=5.4(5.7); % N=11.41(11.59); %
Cl-=4.93(4.89)
Step B:
4-[{[1,2-Dimethyl-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydro-
isoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbon-
yl}(1-methyl-1H-pyrazol-4-yl)amino]phenyl disodium phosphate
[0295] The procedure is in accordance with a protocol analogous to
that described in Steps E and F of Example 1.
[0296] IR (cm.sup.-1): .nu.: C.dbd.O: 1628; .nu.: (phosphate;
ether): 1238, 1143,1113, 985; .gamma.: >CH Ar: 740
[0297] Elemental Microanalysis:
TABLE-US-00003 % C % H % N Calculated 57.64 4.84 10.34 Found 56.62
4.54 10.14
[0298] High-Resolution Mass Spectrometry (ESI+-/FIA/HR):
[0299] Empirical formula:
C.sub.39H.sub.39ClN.sub.6Na.sub.2O.sub.9P
[0300] [M-Na+H].sup.+ calculated: 791.2565
[0301] [M-Na+H].sup.+ measured: 791.2564
EXAMPLE 14
4-[{[1,2-Dimethyl-5-(6-{[(3R)-3-[3-(morpholin-4-yl)propyl]-3,4-dihydroisoq-
uinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbonyl}(-
1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyl disodium
phosphate
Step A:
N-(4-Hydroxyphenyl)-1,2-dimethyl-N-(1-methyl-1H-pyrrolo[2,3-b]pyri-
din-5-yl)-5-(6-{[(3R)-3-[3-(morpholin-4-yl)propyl]-3,4-dihydroisoquinolin--
2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrole-3-carboxamide
hydrochloride
[0302] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compounds of Preparations 1 and 1' used in Step A by the
compounds of Preparations 5 and 3' and, on the other hand, the
compound of Preparation 1'' used in Step C by that of Preparation
3''.
[0303] Elemental Microanalysis: (%, Measured(Theoretical))
[0304] % C=67.63(68.06); % H=5.27(5.95); % N=10.08(10.13); %
Cl-=4.53(4.27)
[0305] High-Resolution Mass Spectrometry (ESI+):
[0306] Empirical formula: C.sub.35H.sub.32ClN.sub.4O.sub.3
[0307] [M+H].sup.+ calculated: 793.3708
[0308] [M+H].sup.+ measured: 793.3704
Step B:
4-[{[1,2-Dimethyl-5-(6-{[(3R)-3-[3-(morpholin-4-yl)propyl]-3,4-dih-
ydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]ca-
rbonyl}(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyl
disodium phosphate
[0309] The procedure is in accordance with a protocol analogous to
that described in Steps E and F of Example 1.
[0310] Unless otherwise mentioned, the compounds of the following
Examples are synthesised in accordance with the process of Example
1 using: (i) the appropriate acid obtained according to one of
Preparations 1 to 9 and (ii) the appropriate tetrahydroisoquinoline
compound obtained according to one of Preparations 1' to 4' and, in
Step C: (iii) the suitable NHR.sub.3R.sub.4 amine (a non-exhaustive
list is proposed in Preparations 1'' to 8'').
EXAMPLE 13
4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin--
2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4--
yl)amino]phenyl disodium phosphate
Step A:
5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinol-
in-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-N-(pyridin-4-
-yl)-1H-pyrrole-3-carboxamide hydrochloride
[0311] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 4 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 4''. The
product obtained is finally subjected to a step of conversion into
salt form in the presence of 1M HCl in ether. After filtration and
lyophilisation in a mixture of acetonitrile/water, the expected
compound is obtained.
[0312] High-Resolution Mass Spectrometry (ESI+):
[0313] Empirical formula: C.sub.39H.sub.38ClN.sub.5O.sub.4
[0314] [M+H].sup.+ calculated: 676.2685
[0315] [M+H].sup.+ measured: 676.2684
Step B:
4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoq-
uinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(py-
ridin-4-yl)amino]phenyl disodium phosphate
[0316] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 16.
[0317] .sup.31P NMR (500 MHz, D.sub.2O) .delta. ppm: -0.05
[0318] IR (cm.sup.-1): .nu.: C.dbd.O: 1631; .nu.: (phosphate;
ether): 1243, 1136, 1112, 982; .gamma.: >CH Ar: 883, 745
[0319] Elemental Microanalysis:
TABLE-US-00004 % C % H % N Calculated 58.54 4.66 8.75 Found 58.23
4.51 8.76
[0320] High-Resolution Mass Spectrometry (ESI+):
[0321] Empirical formula:
C.sub.39H.sub.37ClN.sub.5Na.sub.2O.sub.7P
[0322] [M-Na+2H].sup.+ calculated: 778.2168
[0323] [M-Na+2H].sup.+ measured: 778.2169
EXAMPLE 14
4-[{[5-(5-Fluoro-4-methoxy-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydrois-
oquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(-
1-methyl-1H-pyrazol-4-yl)amino]phenyl disodium phosphate
EXAMPLE 15
4-[{[5-(5-Fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin--
2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1-
H-pyrazol-4-yl)amino]phenyl disodium phosphate
Step A:
5-(5-Fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinol-
in-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-N-(1-methyl--
1H-pyrazol-4-yl)-1H-pyrrole-3-carboxamide hydrochloride
[0324] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 7 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 5''. The
product obtained is finally subjected to a step of conversion into
salt form in the presence of HCl in ether. After filtration and
lyophilisation in a mixture of acetonitrile/water, the expected
compound is obtained.
[0325] Elemental Microanalysis: (%, Measured(Theoretical))
[0326] % C=65.69(65.28); % H=5.38(5.77); % N=11.18(12.02); %
Cl-=5.61(5.07)
Step B:
4-[{[5-(5-Fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoq-
uinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1--
methyl-1H-pyrazol-4-yl)amino]phenyl disodium phosphate
[0327] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 16.
[0328] .sup.31P NMR (400/500 MHz, CD.sub.3OD) .delta. ppm: -0.5
[0329] IR (cm.sup.-1): .nu.: C.dbd.O: 1628; .nu.: (phosphate;
ether): 1238, 1114, 983
[0330] Elemental Microanalysis:
TABLE-US-00005 % C % H % N Calculated 58.02 4.87 10.68 Found 59.03
4.98 10.14
[0331] High-Resolution Mass Spectrometry (ESI+):
[0332] Empirical formula:
C.sub.38H.sub.38FN.sub.6Na.sub.2O.sub.7P
[0333] [M-2Na+3H].sup.+ calculated: 743.2752
[0334] [M-2Na+3H].sup.+ measured: 743.2760
EXAMPLE 16
4-[{[1,2-Dimethyl-5-(7-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquino-
lin-2(1H)-yl]carbonyl}-2,3-dihydro-1,4-benzodioxin-6-yl)-1H-pyrrol-3-yl]ca-
rbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyl disodium
phosphate
Step A:
N-(4-Hydroxyphenyl)-1,2-dimethyl-N-(1-methyl-1H-pyrazol-4-yl)-5-(7-
-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-
-2,3-dihydro-1,4-benzodioxin-6-yl)-1H-pyrrole-3-carboxamide
hydrochloride
[0335] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 8 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 5''. The
product obtained is finally subjected to a step of conversion into
salt form in the presence of 1M HCl in ether. After filtration and
lyophilisation in a mixture of acetonitrile/water, the expected
compound is obtained.
[0336] Elemental Microanalysis: (%, Theoretical:Measured)
[0337] % C=64.99:64.67; % H=5.86:5.67; % N=11.37:11.27; %
Cl-=4.8:4.71
[0338] High-Resolution Mass Spectrometry (ESI+):
[0339] Empirical formula: C.sub.40H.sub.43N.sub.6O.sub.6
[0340] [M+H].sup.+ calculated: 703.3236
[0341] [M+H].sup.+ measured: 703.3239
Step B:
4-[{[1,2-Dimethyl-5-(7-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydro-
isoquinolin-2(1H)-yl]carbonyl}-2,3-dihydro-1,4-benzodioxin-6-yl)-1H-pyrrol-
-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyl
N,N,N',N'-tetramethylphosphorodiamidate
[0342] To a solution of 125 mg of the compound of Step A (0.18
mmol) in dichloromethane (6 mL) there are added 55 .mu.L of
diazabicyclo[5.4.0]undec-7-ene (DBU; 0.36 mmol), and then 33 .mu.L
of bisdimethylaminophosphoryl chloride (0.19 mmol) and 2 mg of
dimethylamino-4-pyridine (0.02 mmol). The reaction mixture is
stirred for 15 hours, diluted with dichloromethane and then with
saturated aqueous sodium carbonate solution. The aqueous phase is
extracted with dichloromethane; the organic phases are then
combined, washed with water and with brine and dried over magnesium
sulphate. After evaporating off the solvents, the crude reaction
product is used directly in the next Step.
Step C:
4-[{[1,2-Dimethyl-5-(7-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydro-
isoquinolin-2(1H)-yl]carbonyl}-2,3-dihydro-1,4-benzodioxin-6-yl)-1H-pyrrol-
-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyl disodium
phosphate
[0343] 4 mL of trifluoroacetic acid are added dropwise to a
solution of 125 mg of the compound of Step B (0.15 mmol) in a 1:1
mixture of acetonitrile and water (5 mL). After stirring for 20
hours at ambient temperature, the reaction mixture is evaporated to
dryness, keeping the temperature of the water bath below 40.degree.
C., and then the residue is treated with a solution of sodium
carbonate (95 mg; 0.9 mmol) in water (4 mL). After stirring for 2
hours at ambient temperature, the reaction mixture is evaporated to
dryness and then 6 mL of anhydrous ethanol are added. The solid is
filtered off, and the filtrate is concentrated to dryness, and then
purified over an OASIS.RTM. phase (acetonitrile/water
gradient).
[0344] .sup.31P NMR (500 MHz, D.sub.2O) .delta. ppm: 0.9
[0345] IR (cm.sup.-1): .nu.: C.dbd.O: 1623; .nu.: (phosphate;
ether): 1235, 1162, 1115, 1065, 985; .gamma.: >CH Ar: 745
[0346] High-Resolution Mass Spectrometry (ESI+):
[0347] Empirical formula:
C.sub.40H.sub.41N.sub.6Na.sub.2O.sub.9P
[0348] [M-2Na+3H].sup.+ calculated: 783.2902
[0349] [M-2Na+3H].sup.+ measured: 783.2907
EXAMPLE 17
5-[{[5-(5-Fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin--
2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4--
yl)amino]pyrimidin-2-yl disodium phosphate
EXAMPLE 18
5-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin--
2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4--
yl)amino]pyrimidin-2-yl disodium phosphate
EXAMPLE 19
4-({[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin--
2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}[1-(trideut-
eriomethyl)-1H-pyrazol-4-yl]amino)phenyl disodium phosphate
Step A:
5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinol-
in-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-N-[1-(trideu-
teriomethyl)-1H-pyrazol-4-yl]-1H-pyrrole-3-carboxamide
hydrochloride
[0350] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 4 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 6''. The
product obtained is finally subjected to a step of conversion into
salt form in the presence of 1M HCl in ether. After filtration and
lyophilisation in a mixture of acetonitrile/water, the expected
compound is obtained.
[0351] Elemental Microanalysis: (%, Theoretical:Measured)
[0352] % C=63.51:63.41; % H=5.63:5.42; % N=11.69:11.61; %
Cl.sup.-=4.93:4.85
[0353] High-Resolution Mass Spectrometry (ESI+-/FIA/HR,
ESI-/FIA):
[0354] Empirical formula:
C.sub.38H.sub.36ClD.sub.3N.sub.6O.sub.4
[0355] [M+H].sup.+ calculated: 682.2982
[0356] [M+H].sup.+ measured: 682.2986
Step B:
4-({[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoq-
uinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}[1--
(trideuteriomethyl)-1H-pyrazol-4-yl]amino)phenyl disodium
phosphate
[0357] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 16.
[0358] .sup.31P NMR (500 MHz, D.sub.2O) .delta. ppm: 4.8
[0359] IR (cm.sup.-1): .nu.: C.dbd.O: 1626; .nu.: (phosphate;
ether): 1243, 1141, 1115, 982; .gamma.: >CH Ar: 880, 831
[0360] High-Resolution Mass Spectrometry (ESI/FIA/HR and
MS/MS):
[0361] Empirical formula:
C.sub.38H.sub.35ClD.sub.3N.sub.6Na.sub.2O.sub.7P
[0362] [M+H].sup.+ calculated: 806.2285
[0363] [M+H].sup.+ measured: 806.2280
EXAMPLE 20
4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin--
2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1,-
2-dimethyl-1H-pyrrol-3-yl)amino]phenyl disodium phosphate
Step A:
5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinol-
in-2(1H)-yl]carbonyl}phenyl)-N-(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)-N-(4--
hydroxyphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxamide
hydrochloride
[0364] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 4 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 7''. The
product obtained is finally subjected to a step of conversion into
salt form in the presence of 1M HCl in ether. After filtration and
lyophilisation in a mixture of acetonitrile/water, the expected
compound is obtained.
[0365] .sup.1H NMR (500 MHz, dmso-d6) .delta. ppm: 11.2 (bs, 1H),
9.39 (bs,1H), 7.83 (d, 1H), 7.54 (d, 1H), 7.33 (s, 1H), 7.14 (m,
2H), 7 (m, 2H), 6.8 (d, 2H), 6.62 (d, 2H), 6.57 (bs, 1H), 5.26 (s,
1H), 5.26 (m, 1H), 4.64/4.03 (AB, 2H), 4.01/3.92 (2m, 4H),
3.75/3.43/3.15/3.02 (4m, 4H), 3.59 (s, 3H), 3.3/3.15 (2m, 2H), 2.97
(s, 3H), 2.69/2.52 (dd+d, 2H), 2.06 (s, 3H), 1.91 (s, 3H)
[0366] Elemental Microanalysis: (%, Theoretical:Measured)
[0367] % C=65.34:65.50; % H=5.62:5.15; % N=11.15:10.84%
Cl-=4.70:4.44
[0368] High-Resolution Mass Spectrometry (ESI+):
[0369] Empirical formula: C.sub.41H.sub.41ClN.sub.6O.sub.4
[0370] [M+H].sup.+ calculated: 717.2952
[0371] [M+H].sup.+ measured: 717.2951
Step B:
4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoq-
uinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5--
cyano-1,2-dimethyl-1H-pyrrol-3-yl)amino]phenyl disodium
phosphate
[0372] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 16.
[0373] .sup.31P NMR (500 MHz, dmso-d6) .delta. ppm: 3.7
[0374] IR (cm.sup.-1): .nu.: --CN: 2210 cm.sup.-1; .nu.: C.dbd.O:
1623; .nu.: (phosphate; ether): 1227, 1133, 1110, 982; .gamma.:
>CH Ar: 884-741
[0375] Elemental Microanalysis:
TABLE-US-00006 % C % H % N Calculated 58.54 4.79 9.99 Found 58.75
4.71 10.18
[0376] High-Resolution Mass Spectrometry (ESI+-/FIA/HR):
[0377] Empirical formula:
C.sub.41H.sub.40ClN.sub.6Na.sub.2O.sub.7P
[0378] [M-2Na+H].sup.+ calculated: 797.2614
[0379] [M-2Na+H].sup.+ measured: 797.2618
EXAMPLE 21
4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin--
2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1--
methyl-1H-pyrrol-3-yl)amino]phenyl disodium phosphate
Step A:
5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinol-
in-2(1H)-yl]carbonyl}phenyl)-N-(5-cyano-1-methyl-1H-pyrrol-3-yl)-N-(4-hydr-
oxyphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxamide hydrochloride
[0380] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 4 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 8''. The
product obtained is finally subjected to a step of conversion into
salt form in the presence of 1M HCl in ether. After filtration and
lyophilisation in a mixture of acetonitrile/water, the expected
compound is obtained.
[0381] Elemental Microanalysis: (%, Theoretical:Measured)
[0382] % C=64.95:65.09; % H=5.45:5.20; % N=11.36:11.26; %
Cl-=4.79:4.62
[0383] High-Resolution Mass Spectrometry (ESI/+):
[0384] Empirical formula: C.sub.40H.sub.39ClN.sub.6O.sub.4
[0385] [M+H].sup.+ calculated: 703.2794
[0386] [M+H].sup.+ measured: 703.2789
Step B
4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoqu-
inolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-c-
yano-1-methyl-1H-pyrrol-3-yl)amino]phenyl disodium phosphate
[0387] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 16.
[0388] .sup.31P NMR (500 MHz, dmso-d6) .delta. ppm: 4.5
[0389] IR (cm.sup.-1): .nu.: --CN: 2215 cm.sup.-1; .nu.: C.dbd.O
1626; .nu.: (Phosphate; ether): 1227, 1141, 1112, 982; .gamma.
>CH Ar: 826-742
[0390] High-Resolution Mass Spectrometry (ESI+-/FIA/HR):
[0391] Empirical formula:
C.sub.40H.sub.38ClN.sub.6Na.sub.2O.sub.7P
[0392] [M-2Na+3H].sup.+ calculated: 783.2457
[0393] [M-2Na+3H].sup.+ measured: 783.2462
EXAMPLE 22
4-[{[3-(6-{[(3R)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]-
carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}-
(phenyl)amino]phenyl disodium phosphate
Step A:
N-(4-Hydroxyphenyl)-3-{6-[((3R)-3-(4-morpholinylmethyl)-3,4-dihydr-
o-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tet-
rahydro-1-indolizine carboxamide hydrochloride
[0394] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing the compound of
Preparation 1' used in Step A by the compound of Preparation 4'.
The solid is then dissolved in dichloromethane, and 2 mL of 1M HCl
in ether are added. The batch is stirred for 1 hour and then
evaporated to dryness. The hydrochloride thereby obtained is
dissolved in a mixture of water/acetonitrile until dissolution is
complete, and then lyophilised.
[0395] Elemental Microanalysis:
TABLE-US-00007 % C % H % N % Cl.sup.- Calculated 69.11 5.80 7.50
4.74 Found 68.89 5.23 7.41 4.62
[0396] Optical rotation: (.alpha.).sub.D.sup.20=-45.1.degree. (c=9
mg/mL, MeOH)
Step B:
4-[{[3-(6-{[(3R)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2-
(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]c-
arbonyl}(phenyl)amino]phenyl disodium phosphate
[0397] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 16.
[0398] .sup.31P NMR (400/500 MHz, dmso-d6) .delta. ppm: 2.6
[0399] Elemental Microanalysis:
TABLE-US-00008 % C % H % N Calculated 61.87 4.95 6.71 Found 61.33
4.93 7.14
[0400] High-Resolution Mass Spectrometry (ESI+-/FIA/HR):
[0401] Empirical formula:
C.sub.43H.sub.41N.sub.4Na.sub.2O.sub.9P
[0402] [M-2Na+H[.sup.+ calculated: 791.2840
[0403] [M-2Na+H[.sup.+ measured: 791.2845
EXAMPLE 23
4-[(1-Methyl-2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-5-yl){[3-(2-{[(3S)-3-(mo-
rpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-4-[2-oxo-2-(-
piperidin-1-yl)ethoxy]phenyl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}am-
ino]phenyl disodium phosphate
Step A: Methyl
3-[4-benzyloxy-2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-
-carbonyl]phenyl]-5,6,7,8-tetrahydroindolizine-1-carboxylate
[0404] To a solution of 14.19 g (35.0 mmol) of the compound
obtained in Preparation 9 in 200 mL of dimethylformamide there are
successively added
4-[[(3S)-1,2,3,4-tetrahydroisoquinolin-3-yl]methyl]morpholine
(Preparation 1'; 8.13 g; 35.0 mmol), hydroxybenzotriazole (6.15 g;
45.5 mmol), 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide
hydrochloride (6.70 g; 45.5 mmol) and triethylamine (21.95 mL; 0.16
mol). The batch is then stirred overnight at ambient temperature.
The reaction mixture is then poured into 400 mL of ethyl acetate
and then successively washed with saturated aqueous NaHCO.sub.3
solution, water and brine. The combined aqueous phases are
extracted with ethyl acetate. The resulting organic phases are
dried over sodium sulphate, filtered and concentrated under reduced
pressure. The product obtained is purified by chromatography over
silica gel to provide the title compound.
[0405] .sup.1H NMR (500 MHz, dmso-d6, 300K) .delta. ppm: 7.5-7.3
(m, 5H), 7.38 (d, 1H), 7.2-6.9 (m, 4H), 7.15 (dd, 1H), 6.9 (d, 1H),
6.35/6.25/6.08 (3*s, 1H), 5.21/5.12 (3*s, 2H), 5.09/4.85/3.7 (3*m,
1H), 4.9-3.8 (8*d, 2H), 4.2-3.4 (m, 2H), 3.65/3.6/3.55 (3*s, 3H),
3.6-3.4 (m, 4H), 3-2.4 (m, 2H), 2.9-1.8 (6*dd, 2H), 2.5-1.95 (4*m,
4H), 2.35-1.7 (6*m, 2H), 2-1.45 (6*m, 4H)
Step B:
3-[4-Benzyloxy-2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquin-
oline-2-carbonyl]phenyl]-5,6,7,8-tetrahydroindolizine-1-carboxylic
acid
[0406] 40.1 mL of 1M aqueous LiOH solution are added to a solution
of 12.7 g (20 mmol) of the compound obtained in the Step above in
40 mL of dioxane. The batch is heated at 100.degree. C. overnight.
The reaction mixture is poured into water and then extracted with
ethyl ether. The ethereal phase is extracted once more with water.
The combined aqueous phases are acidified to pH 4 by adding
powdered citric acid, and then extracted with dichloromethane. The
dichloromethane phase is washed with brine, dried over sodium
sulphate, filtered and concentrated to dryness. The title product
is obtained in the form of a meringue.
[0407] .sup.1H NMR (500 MHz, dmso-d6, 300K) .delta. ppm: 11.35 (bs,
1H), 7.5-7.3 (m, 5H), 7.38 (m, 1H), 7.2-6.9 (m, 4H), 7.15 (m, 1H),
6.9 (m, 1H), 6.31/6.25/6.1 (3*s, 1H), 5.22/5.2/5.15 (3*s, 2H),
5.1/4.82/3.7 (3*m, 1H), 4.85-3.8 (8*d, 2H), 4.2-3.4 (m, 2H),
3.6-3.45 (m, 4H), 3-2.3 (m, 2H), 2.9-1.8 (m, 2H), 2.5-1.9 (6*m,
4H), 2.35-1.8 (6*m, 2H), 1.9-1.3 (m, 4H)
Step C:
3-[4-Benzyloxy-2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquin-
oline-2-carbonyl]phenyl]-N-[4-[tert-butyl(dimethyl)silyl]oxyphenyl]-N-(1-m-
ethylpyrrolo[2,3-b]pyridin-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamid-
e
[0408] The acid obtained in Step B (9 g, 11.8 mmol) is dissolved in
90 mL of 1,2-dichloroethane. 1.9 mL of
1-chloro-N,N,2-trimethylpropenylamine (14 mmol) are added thereto.
After stirring for 3 hours at ambient temperature, 90 mL of toluene
and 4.62 g of
N-[4-[tert-butyl(dimethyl)silyl]oxyphenyl]-1-methyl-1H-pyrrolo[2,3-b]pyri-
din-5-amine (Preparation 3'', 13 mmol) are added. The reaction
mixture is heated at 110.degree. C. for 20 hours. After returning
to ambient temperature, the reaction mixture is washed with brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness.
The residue obtained is purified by chromatography over silica gel
(dichloromethane/ethanol gradient) to yield the expected
product.
[0409] .sup.1H NMR (500 MHz, dmso-d6, 300K) .delta. ppm:
7.95/7.8/7.75 (3*d, 1H), 7.68/7.65/7.4 (3*d, 1H), 7.4/7.3 (2*d,
1H), 7.25-6.8 (m, 9H), 7.05/6.9 (2*m, 1H), 7-6.6 (3*bd, 2H), 6.9
(m, 1H), 6.75-6.45 (3*bd, 2H), 6.7 (m, 1H), 6.3 (2*d, 1H),
5.15-4.95 (m, 2H), 5.15/5.1/4.8 (3*s, 1H), 4.95/4.6/3.5 (3*m, 1H),
4.9-3.7 (8*d, 2H), 3.8-3.3 (3*m, 2H), 3.75/3.7/3.5 (3*s, 3H),
3.45/3.3 (2*m, 4H), 3-2.5 (3*m, 2H), 3-2.3 (m, 2H), 2.4-1.75 (5*m,
4H), 2.25-1.7 (6*m, 2H), 1.75-1.3 (m, 4H), 0.7 (bs, 9H), 0.1 (m,
6H)
Step D:
N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-3-[4-hydroxy-2-[(3S)-3--
(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]phenyl]-N-(1-met-
hylpyrrolo[2,3-b]pyridin-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide
[0410] 0.9 g of Pd/C 10% is added to a solution, in 100 mL of
ethanol, of 8.88 g (8.4 mmol) of the compound obtained in Step C,
whilst bubbling through argon. The reaction mixture is placed under
1.2 bars of hydrogen at ambient temperature for 15 hours. The
catalyst is filtered off and the solvent is evaporated off under
reduced pressure to provide the title compound.
[0411] .sup.1H NMR (500 MHz, dmso-d6, 300K) .delta. ppm:
8.06/7.92/7.87 (3*d, 1H), 7.75/7.5/7.39 (3*d, 1H), 7.5 (m, 1H),
7.28-6.9 (m, 5H), 6.87/6.7 (2*m, 2H), 6.76 (m, 1H), 6.75/6.67/6.62
(3*m, 2H), 6.67/6.46 (m, 1H), 6.4/6.36 (2*m, 1H), 5.19/5.13/4.9
(3*bs, 1H), 5.06/4.7/3.6 (3*m, 1H), 4.97/4.2/4.15/4.07 (4*m, 2H),
4.87/4.81 (bs, 1H), 3.86/3.56/3.39 (3*m, 2H), 3.78/3.57 (2*m, 3H),
3.59/3.44 (2*m, 4H), 2.96-2.61 (2*m, 2H), 2.88/2.6 (2*m, 2H),
2.59-1.81 (m, 6H), 1.87-1.42 (m, 4H), 0.89 (s, 9H), 0.12 (m,
6H)
Step E:
N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-N-(1-methylpyrrolo[2,3--
b]pyridin-5-yl)-3-[2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinolin-
e-2-carbonyl]-4-[2-oxo-2-(1-piperidyl)ethoxy]phenyl]-5,6,7,8-tetrahydroind-
olizine-1-carboxamide
[0412] The compound of Step D (3.0 g, 2.9 mmol) is dissolved in 100
mL of toluene. 1.53 g (5.8 mmol) of triphenylphosphine and 0.62 g
(4.3 mmol) of 2-hydroxy-1-(1-piperidyl)ethanone are added thereto.
The mixture is heated to 50.degree. C., and then 1.01 g (4.3 mmol)
of di-tert-butyl azodicarboxylate are added. The reaction mixture
is stirred at 50.degree. C. for 1 hour and is then allowed to
return to ambient temperature before adding 1 mL of trifluoroacetic
acid. After stirring overnight at ambient temperature, the mixture
is successively washed with water, saturated NaHCO.sub.3 and brine
solution. The combined aqueous phases are extracted with ethyl
acetate. The resulting organic phases are dried over sodium
sulphate, filtered and concentrated under reduced pressure. The
crude product obtained is purified by chromatography over silica
gel (dichloromethane/ethanol 98/2) to yield the expected
compound.
[0413] .sup.1H NMR (500 MHz, dmso-d6, 300K) .delta. ppm:
8.06/7.92/7.87 (3*d, 1H), 7.75/7.51/7.4 (3*d, 1H), 7.49 (2*d, 1H),
7.29-6.89 (m, 5H), 6.93 (m, 1H), 6.88/6.7 (m, 2H), 6.75/6.67 (m,
1H), 6.75/6.68/6.59 (3*m, 2H), 6.4/6.36 (2*m, 1H), 5.2/5.16/4.92
(3*m, 1H), 5.06/4.69/3.58 (3*m, 1H), 4.97/4.25/4.16/4.03 (4*d, 2H),
4.89/4.81 (2*m, 2H), 3.79/3.59 (2*m, 3H), 3.59/3.43/3.4 (3*m, 6H),
3.58/3.43 (2*m, 4H), 3.03-2.61 (m, 2H), 2.97-2.65 (m, 2H),
2.57-1.74 (m, 6H), 1.89-1.3 (m, 10H), 0.89 (2bs, 9H), 0.11 (m,
6H)
Step F:
N-(4-Hydroxyphenyl)-N-(1-methylpyrrolo[2,3-b]pyridin-5-yl)-3-[2-[(-
3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-4-[2-oxo--
2-(1-piperidyl)ethoxy]phenyl]-5,6,7,8-tetrahydroindolizine-1-carboxamide
[0414] A 1M solution of tetrabutylammonium fluoride (3.14 mL, 3
mmol) in tetrahydrofuran is added at ambient temperature to a
solution, in 30 mL of tetrahydrofuran, of the compound obtained in
Step E (2.92 g, 2.9 mmol). After stirring for 5 minutes, the
reaction mixture is poured into a 50/50 mixture of ethyl acetate
and saturated aqueous NaHCO.sub.3 solution. The separated organic
phase is washed with water and then with brine. The combined
aqueous phases are extracted with ethyl acetate. The resulting
organic phases are dried over sodium sulphate, filtered and
concentrated under reduced pressure. The crude product obtained is
purified by chromatography over silica gel
(dichloromethane/ethanol/ammonia gradient) to yield the title
compound.
[0415] .sup.1H NMR (500 MHz, dmso-d6, 300K) .delta. ppm: 9.4 (m,
OH), 8.1-7.8 (3*d, 1H), 7.7-7.3 (2*m, 1H), 7.5/7.4 (2*m, 1H),
7.3-6.9 (m, 4H), 7.2 (m, 1H), 6.9 (m, 1H), 6.8-6.5 (m, 2H), 6.7-6.5
(m, 2H), 6.7 (m, 1H), 6.4 (m, 1H), 5.3-5 (m, 1H), 5.1/4.7/3.6 (3*m,
1H), 5-3.6 (m, 2H), 5-3.6 (m, 2H), 4.8 (m, 2H), 3.8-3.6 (m, 3H),
3.6/3.4 (m, 2H), 3.4 (m, 6H), 3.1-2.5 (m, 2H), 2.9-1.9 (m, 2H),
2.5-1.7 (m, 4H), 1.8-1.4 (m, 6H), 1.6-1.3 (m, 4H)
Step G:
N-(4-Hydroxyphenyl)-N-(1-methyl-2,3-dihydropyrrolo[2,3-b]pyridin-5-
-yl)-3-[2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbony-
l]-4-[2-oxo-2-(1-piperidyl)ethoxy]phenyl]-5,6,7,8-tetrahydroindolizine-1-c-
arboxamide
[0416] 0.71 g (11 mmol) of sodium cyanoborohydride is added to a
solution, in 20 mL of acetic acid, of the compound obtained in Step
F (2.0 g, 2.2 mmol). After stirring for 14 hours at ambient
temperature, 0.36 g (5.5 mmol) of sodium cyanoborohydride is again
added, and then the reaction mixture is heated at 50.degree. C. for
3 hours before a second addition of 0.1 eq. of sodium
cyanoborohydride to complete the reaction in 30 minutes at
50.degree. C. The acetic acid is evaporated off under reduced
pressure, and then the residue is taken up in dichloromethane and
washed with saturated aqueous NaHCO.sub.3 solution, water and
brine. The combined aqueous phases are extracted with
dichloromethane. The resulting organic phases are dried over sodium
sulphate, filtered and concentrated under reduced pressure. The
crude product obtained is purified by chromatography over silica
gel (dichloromethane/ethanol/ammonia gradient) to yield the title
compound in the form of a meringue.
[0417] .sup.1H NMR (500 MHz, dmso-d6, 300K) .delta. ppm: 9.3 (bs,
1H), 7.5/7.4/7.3 (3*m, 1H), 7.2/6.7 (2*m, 1H), 7.2 (m, 1H), 7.1-6.8
(m, 4H), 6.9/6.7 (m, 1H), 6.9 (m, 1H), 6.8-6.5 (m, 2H), 6.7-6.5 (m,
2H), 5.3-5.1 (2*d, 1H), 5.1/4.7/3.6 (3*m, 1H), 4.9/4.2-3.5 (2*m,
1H), 4.9/4.2-3.5 (2*, 1H), 4.9-4.8 (m, 2H), 3.6/3.4 (2*m, 4H),
3.4/3.3 (m, 2H), 3.4 (m, 6H), 3.1-2.5 (m, 4H), 3-2.4 (m, 2H),
2.8/2.6 (m, 3H), 2.6-1.7 (m, 6H), 1.9-1.3 (m, 10H)
Step H:
N-(4-Hydroxyphenyl)-N-(1-methyl-2,3-dihydropyrrolo[2,3-b]pyridin-5-
-yl)-3-[2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbony-
l]-4-[2-oxo-2-(1-piperidyl)ethoxy]phenyl]-5,6,7,8-tetrahydroindolizine-1-c-
arboxamide hydrochloride
[0418] The base obtained in Step G (0.60 g, 0.69 mmol) is dissolved
in acetonitrile and then converted into salt form using 0.7 mL (0.7
mmol) of 1N HCl solution. The solution is filtered, frozen and then
lyophilised to provide the title compound in the form of a
powder.
[0419] Elemental Microanalysis: (%, Theoretical:Measured)
[0420] % C=68.02:68.06; % H=6.49:6.21; % N=10.89:10.87; %
Cl=4.14:3.94
[0421] High-Resolution Mass Spectrometry (ESI+):
[0422] Empirical formula: C.sub.51H.sub.57N.sub.7O.sub.6
[0423] [M+H].sup.+ calculated: 864.4445
[0424] [M+H].sup.+ measured: 864.4443
Step I:
4-[(1-Methyl-2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-5-yl){[3-(2-{[(3-
S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-4-[2-
-oxo-2-(piperidin-1-yl)ethoxy]phenyl)-5,6,7,8-tetrahydroindolizin-1-yl]car-
bonyl}amino]phenyl disodium phosphate
[0425] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 16.
[0426] IR (cm.sup.-1): .nu.: C.dbd.O: 1625; .nu.: (phosphate;
ether): 1229, 1138, 1115, 982; .gamma.: >CH Ar: 880-748-745
[0427] Elemental Microanalysis:
TABLE-US-00009 % C % H % N Calculated 62.00 5.71 9.92 Found 61.45
5.53 9.96
[0428] High-Resolution Mass Spectrometry (ESI+):
[0429] Empirical formula:
C.sub.51H.sub.56N.sub.7Na.sub.2O.sub.9P
[0430] [M-2Na+3H].sup.+ calculated: 944.4106
[0431] [M-2Na+3H].sup.+ measured: 944.4116
EXAMPLE 24
4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin--
2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1-
H-pyrazol-4-yl)amino]phenyl disodium phosphate
Step A:
5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinol-
in-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-N-(1-methyl--
1H-pyrazol-4-yl)-1H-pyrrole-3-carboxamide hydrochloride
[0432] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 4 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 5''. The
product obtained is finally subjected to a step of conversion into
salt form in the presence of HCl in ether. After filtration and
lyophilisation in a mixture of acetonitrile/water, the expected
compound is obtained.
[0433] Elemental Microanalysis: (%, Theoretical:Measured)
[0434] % C=63.77:62.83; % H=5.63:5.83; % N=11.74:11.29; %
Cl-=4.95:5.42
Step B:
4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoq-
uinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1--
methyl-1H-pyrazol-4-yl)amino]phenyl disodium phosphate
[0435] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 16.
[0436] IR (cm.sup.-1): .nu.: C.dbd.O: 1625; .nu.: (phosphate;
ether): 1241, 1146, 1112, 983
[0437] Elemental Microanalysis:
TABLE-US-00010 % C % H % N Calculated 56.83 4.77 10.46 Found 56.82
4.58 10.43
[0438] High-Resolution Mass Spectrometry (ESI+):
[0439] Empirical formula:
C.sub.38H.sub.38ClN.sub.6Na.sub.2O.sub.7P
[0440] [M-2Na+3H].sup.+ calculated: 759.2457
[0441] [M-2Na+3H].sup.+ measured: 759.2465
EXAMPLE 25
4-[(5-Cyano-1,2-dimethyl-1H-pyrrol-3-yl){[5-(5-fluoro-2-{[(3S)-3-(morpholi-
n-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethy-
l-1H-pyrrol-3-yl]carbonyl}amino]phenyl disodium phosphate
Step A:
N-(5-Cyano-1,2-dimethyl-1H-pyrrol-3-yl)-5-(5-fluoro-2-{[(3S)-3-(mo-
rpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(4--
hydroxyphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxamide
hydrochloride
[0442] The procedure is in accordance with a protocol analogous to
that described in Steps A-D of Example 1 replacing, on the one
hand, the compound of Preparation 1 used in Step A by the compound
of Preparation 7 and, on the other hand, the compound of
Preparation 1'' used in Step C by that of Preparation 7''. The
product obtained is finally subjected to a step of conversion into
salt form in the presence of HCl in ether. After filtration and
lyophilisation in a mixture of acetonitrile/water, the expected
compound is obtained.
[0443] High-Resolution Mass Spectrometry (ESI/FIA/HR and
MS/MS):
[0444] Empirical formula: C.sub.41H.sub.41FN.sub.6O.sub.4
[0445] [M+H].sup.+ calculated: 701.3246
[0446] [M+H].sup.+ measured: 701.3282
Step B:
4-[(5-Cyano-1,2-dimethyl-1H-pyrrol-3-yl){[5-(5-fluoro-2-{[(3S)-3-(-
morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-
-dimethyl-1H-pyrrol-3-yl]carbonyl}amino]phenyl disodium
phosphate
[0447] The procedure is in accordance with a protocol analogous to
that described in Steps B and C of Example 16.
[0448] .sup.31P NMR (400/500 MHz, CD.sub.3OD) .delta. ppm: -0.5
[0449] IR (cm.sup.-1): .nu.: --CN: 2211 cm.sup.-1; .nu.: C.dbd.O:
1629; .nu.: (phosphate; ether): 1236, 1114, 984
[0450] Elemental Microanalysis:
TABLE-US-00011 % C % H % N Calculated 59.71 4.89 10.19 Found 60.09
4.95 9.88
[0451] High-Resolution Mass Spectrometry (ESI+):
[0452] Empirical formula:
C.sub.41H.sub.40FN.sub.6Na.sub.2O.sub.7P
[0453] [M-2Na+3H].sup.+ calculated: 781.2909
[0454] [M-2Na+3H].sup.+ measured: 781.2898
Pharmacological and Pharmacokinetic Studies
[0455] For clarification purposes, and in anything which follows,
compounds of formula (I') will be referred to as "drug of Example
x" from which they have been derived. As for an example,
N-(4-hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-
-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-
-1-indolizine carboxamide will be referred to as the "drug from
Example 1".
EXAMPLE A1
Induction of Caspase Activity in Vivo by Compounds of Formula
(I')
[0456] The ability of the compounds of formula (I') to activate
caspase 3 is evaluated in a xenograft model of RS4;11 leukaemia
cells.
[0457] 1.times.10.sup.7 RS4;11 cells are grafted sub-cutaneously
into immunosuppressed mice (SCID strain). 25 to 30 days after the
graft, the animals are treated orally with the various compounds.
Sixteen hours after treatment, the tumour masses are recovered and
lysed, and the caspase 3 activity is measured in the tumour
lysates.
[0458] This enzymatic measurement is carried out by assaying the
appearance of a fluorigenic cleavage product (DEVDase activity,
Promega). It is expressed in the form of an activation factor
corresponding to the ratio between the two caspase activities: the
activity for the treated mice divided by the activity for the
control mice.
[0459]
N-(4-Hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-
-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetr-
ahydro-1-indolizine carboxamide (also referred to as the drug from
Example 1) was tested. At a dose of 100 mg/kg p.o., the in vivo
caspase activation factor is 29.3.
[0460] The results obtained show that the compounds of formula (I')
are capable of inducing apoptosis in RS4;11 tumour cells in
vivo.
EXAMPLE A2
Quantification of the Cleaved Form of Caspase 3 in Vivo Brought
About by Compounds of formula (I')
[0461] The ability of the compounds of formula (I') to activate
caspase 3 is evaluated in a xenograft model of RS4;11 leukaemia
cells.
[0462] 1.times.10.sup.7 RS4;11 cells are grafted sub-cutaneously
into immunosuppressed mice (SCID strain). 25 to 30 days after the
graft, the animals are treated orally with the various compounds.
After treatment, the tumour masses are recovered and lysed, and the
cleaved (activated) form of caspase 3 is quantified in the tumour
lysates.
[0463] The quantification is carried out using the "Meso Scale
Discovery (MSD) ELISA platform" test, which specifically assays the
cleaved form of caspase 3. It is expressed in the form of an
activation factor corresponding to the ratio between the quantity
of cleaved caspase 3 in the treated mice divided by the quantity of
cleaved caspase 3 in the control mice.
[0464] The results show that the compounds of formula (I') are
capable of inducing apoptosis in RS4;11 tumour cells in vivo.
TABLE-US-00012 TABLE 1 Caspase activation factors (cleaved caspase
3 MSD test in the tumours of treated mice versus control mice) in
vivo, after treatment by the oral route Dose Sampling Activation
factor +/- Compound tested (mg/kg) time S.E.M. Drug from Example 13
12.5 2 hours 24.5 +/- 7.5 Drug from Example 19 12.5 2 hours 13.5
+/- 1.2 Drug from Example 20 12.5 2 hours 52.0 +/- 8.6 Drug from
Example 21 12.5 2 hours 22.6 +/- 2.4 Drug from Example 24 25 2
hours 45.7 +/- 2.0 Drug from Example 25 12.5 2 hours 38.7 +/- 10.7
Drug from Example 15 25 2 hours 29.8 +/- 4.0
EXAMPLE A3
Quantification of the Cleaved Form of Caspase 3 in Vivo Brought
About by Compounds of Formula (I)
[0465] The ability of the compounds of formula (I) to activate
caspase 3 is evaluated in a xenograft model of RS4;11 leukaemia
cells in accordance with the protocol given in Example A2.
TABLE-US-00013 TABLE 2 Caspase activation factors (cleaved caspase
3 MSD test in the tumours of treated mice versus control mice) in
vivo, after treatment by the oral route Compound Dose Sampling
Activation tested (mg/kg) time factor +/- S.E.M. Example 13 12.5 2
hours 58.6 +/- 4.6 Example 1 50 2 hours 21.2 +/- 1.3 Example 19
12.5 2 hours 27.5 +/- 3.5 Example 20 12.5 2 hours 62.1 +/- 3.4
Example 21 25 2 hours 55.2 +/- 6.2 Example 24 25 2 hours 60.5 +/-
4.5 Example 25 12.5 2 hours 61.8 +/- 8.9 Example 15 25 2 hours 12.1
+/- 1.1
EXAMPLE B
Solubility of Compounds of Formula (I)
[0466] The solubility of compounds of formula (I) in water was
measured and compared with that of compounds of formula (I').
[0467] More specifically,
4-[{[3-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl-
]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl-
}(phenyl)amino]phenyl disodium phosphate (also referred to as the
compound of Example 1) was tested and compared to
N-(4-hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-
-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-
-1-indolizine carboxamide (also referred to as the drug from
Example 1).
[0468] The solubility of the compound of Example 1 in water is
greater than or equal to 10 mg/mL (12.6mM) whereas that of the
associated drug is only 40 .mu.g/mL (56.2 .mu.M). The solubilities
of the compounds were also measured in a medium buffered to
physiological pH (cf. Table 3).
TABLE-US-00014 TABLE 3 Solubilities in aqueous medium (buffer
solution: phosphate 0.33M, pH = 7.4) of compounds of formula (I)
and the associated compounds of formula (I'), measured at four
concentrations: 10 .mu.M, 20 .mu.M, 50 .mu.M and 100 .mu.M
Solubility at Solubility Solubility Solubility Compound tested 10
.mu.M at 20 .mu.M at 50 .mu.M at 100 .mu.M Example 19 Soluble
Soluble Soluble Soluble Drug from Example 19 Soluble Soluble
Soluble Insoluble Example 20 Soluble Soluble Soluble Soluble Drug
from Example 20 Soluble Insoluble Insoluble Insoluble Example 25
Soluble Soluble Soluble Soluble Drug from Example 25 Soluble
Soluble Insoluble Insoluble Example 1 Soluble Soluble Soluble
Soluble Drug from Example 1 Insoluble Insoluble Insoluble
Insoluble
[0469] The results show that the compounds of formula (I) are much
more soluble than the compounds of formula (I'). Only compounds of
formula (I) exhibit solubilities greater than or equal to 100
.mu.M.
EXAMPLE C
In Vivo Conversion of Compounds of Formula (I)
[0470] The pharmacokinetic profile of the phosphate compounds of
formula (I) is evaluated in a lipid formulation and in aqueous
solution in female SCID mice. This is compared to the
pharmacokinetic profile of compounds of formula (I') in a lipid
formulation. More specifically,
4-[{[3-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl-
]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl-
}(phenyl)amino]phenyl disodium phosphate (also referred to as the
compound of Example 1) was tested and compared to
N-(4-hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-
-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-
-1-indolizine carboxamide (also referred to as the drug from
Example 1).
[0471] Lipid Formulation of the Compound of Example 1
[0472] The compound of Example 1 is prepared in a mixture of
anhydrous ethanol/polyethylene glycol 300/water (10/40/50, v/v/v)
intended for administration by the p.o. route. The study is carried
out in 2 groups of SCID mice to which the compound of Example 1 is
administered under the following conditions: [0473] Group 1: 3
mg/kg p.o. (gavage, 10 mL/kg), [0474] Group 2: 25 mg/kg p.o.
(gavage, 10 mL/kg).
[0475] Blood samples are taken at the following points in time (3
samples per animal and 3 animals for each point in time): 0.25 hr,
0.5 hr, 1 hr, 2 hrs, 6 hrs and 24 hrs after oral
administration.
[0476] Aqueous Formulation of the Compound of Example 1
[0477] The compound of Example 1 is also administered by the oral
route in an aqueous medium to SCID mice, under the following
conditions: [0478] Group 1: 30 mg/kg p.o. dissolved in 1 mM sodium
carbonate solution (gavage, 10 mL/kg), [0479] Group 2: 100 mg/kg
p.o. in water (gavage, 10 mL/kg).
[0480] Blood samples are taken at the following points in time (3
animals for each point in time and 1 sample per animal): 0.25 hr,
0.5 hr, 1 hr, 2 hrs, 4 hrs, 6 hrs and 24 hrs after oral
administration.
[0481] For all formulations of the compound of Example 1, the blood
thereby collected is centrifuged and the plasma is transferred to
tubes containing 1M hydrochloric acid. The plasma concentrations of
the phosphate compound (prodrug) and its hydroxylated homologue
(drug) are determined simultaneously using a method of liquid
chromatography coupled with mass spectrometric detection
(TFC-LC-MS/MS). The limit of detection for both entities is 0.5
ng/mL.
[0482] Lipid Formulation of the Drug from the Compound of Example
1
[0483] The drug from Example 1 is prepared in a mixture of
anhydrous ethanol/polyethylene glycol 300/water (10/40/50, v/v/v)
intended for administration by the p.o. route. The study is carried
out in several groups of SCID mice to which the drug from Example 1
is administered under the following conditions: [0484] Group 1: 3
mg/kg p.o. (gavage, 10 mL/kg), [0485] Group 2: 30 mg/kg p.o.
(gavage, 10 mL/kg), [0486] Group 3: 25 mg/kg p.o. (gavage, 10
mL/kg), [0487] Group 4: 100 mg/kg p.o. (gavage, 10 mL/kg).
[0488] Blood samples are taken at the following points in time (3
animals for each point in time and 3 samples per animal for Groups
1-2 and 1 sample per animal for Groups 3 and 4): [0489] p.o.:
before administration and then 0.25 hr, 0.5 hr, 0.75 hr, 1 hr, 2
hrs, 4 hrs, 6 hrs, 8 hrs, 16 hrs and 24 hrs after oral
administration at 3 and 30 mg/kg, [0490] p.o.: 0.5 hr, 2 hrs, 6
hrs, 16 hrs and 24 hrs after oral administration at 25 mg/kg and
0.5 hr, 1 hr, 2 hrs, 6 hrs, 16 hrs, 30 hrs and 48 hrs after oral
administration at 100 mg/kg.
[0491] The plasma from the blood samples collected after
administration of the lipid formulations of the drug from the
compound of Example 1 are analysed by liquid chromatography coupled
with mass spectrometric detection. The limit of quantification of
the drug from the compound of Example 1 is less than or equal to
0.5 ng/mL.
[0492] Non-compartmental pharmacokinetic analysis is carried out on
the mean values of the plasma concentrations of the compounds
tested. The results are shown in Tables 4 and 5 hereinbelow.
[0493] The results show that, whatever the dose (from 3 to 100
mg/kg) and the carrier (lipid or aqueous formulation), the major
part of the prodrug of formula (I) is rapidly converted in vivo
into the corresponding drug of formula (I') (see Table 4). Plasma
exposure of the prodrug (C.sub.max, AUC) is low in comparison to
that of the corresponding drug. The results also show that the
plasma concentration of the drug so measured (after administration
of the prodrug) is equivalent to or even greater than that measured
after direct administration of the drug by the oral route (see
Table 5).
TABLE-US-00015 TABLE 4 Compound Compound measured administered
Example 1 Drug from Example 1 Example 1, 3 C.sub.max (ng/mL) = 16
C.sub.max (ng/mL) = 342 mg/kg p.o. Lipid T.sub.max (h) = 0.25
T.sub.max (h) = 0.25 formulation AUC.sub.t (ng h/mL) = 5 AUC.sub.t
(ng h/mL) = 314 Example 1, 25 C.sub.max (ng/mL) = 244 C.sub.max
(ng/mL) = 6204 mg/kg p.o. Lipid T.sub.max (h) = 0.25 T.sub.max (h)
= 0.5 formulation AUC.sub.t (ng h/mL) = 92 AUC.sub.t (ng h/mL) =
20952 Example 1, 30 C.sub.max (ng/mL) = 391 C.sub.max (ng/mL) =
11967 mg/kg p.o. T.sub.max (h) = 1.0 T.sub.max (h) = 0.5 Aqueous
AUC.sub.t (ng h/mL) = 879 AUC.sub.t (ng h/mL) = 49416 formulation
Example 1, 100 C.sub.max (ng/mL) = 359 C.sub.max (ng/mL) = 28066
mg/kg p.o. T.sub.max (h) = 2.0 T.sub.max (h) = 2.0 Aqueous
AUC.sub.t (ng h/mL) = 797 AUC.sub.t (ng h/mL) = 168478
formulation
TABLE-US-00016 TABLE 5 Compound measured Compound administered Drug
from Example 1 Drug from Example 1, 3 mg/kg p.o. C.sub.max (ng/mL)
= 295 Lipid formulation T.sub.max (h) = 0.25 AUC.sub.t (ng h/mL) =
225 Drug from Example 1, 25 mg/kg p.o. C.sub.max (ng/mL) = 5070
Lipid formulation T.sub.max (h) = 2.0 AUC.sub.t (ng h/mL) = 20400
Drug from Example 1, 30 mg/kg p.o. C.sub.max (ng/mL) = 8580 Lipid
formulation T.sub.max (h) = 1.0 AUC.sub.t (ng h/mL) = 24200 Drug
from Example 1, 100 mg/kg p.o. C.sub.max (ng/mL) = 25878 Lipid
formulation T.sub.max (h) = 0.5 AUC.sub.t (ng h/mL) = 148046
[0494] More specifically, p.o. administration of the prodrug in an
aqueous carrier makes it possible to obtain plasma concentrations
of the drug which are equivalent to or even greater than those
obtained after direct p.o. administration of the drug in a lipid
carrier. The prodrug therefore offers the benefit of ease of
formulation compared to the corresponding drug, especially in an
aqueous medium, which is very advantageous with a view to clinical
development. Indeed, as Example D shows, the drug from Example 1 is
difficult to formulate in an aqueous medium.
[0495] Aqueous Formulation of the Compounds of Examples 20 and
25
[0496] The compounds of Examples 20 and 25 are administered by the
oral route in an aqueous medium to SCID mice, under the following
conditions: [0497] Group 1: 3 mg/kg p.o. in solution in 1M sodium
carbonate (gavage, 10 mL/kg), [0498] Group 2: 25 mg/kg p.o. in
solution in 1M sodium carbonate (gavage, 10 mL/kg).
[0499] Blood samples are taken at the following points in time (3
animals for each point in time): 0.25 hr, 0.5 hr, 1 hr, 2 hrs, 6
hrs and 24 hrs after oral administration.
[0500] The blood thereby collected is centrifuged and the plasma is
transferred to tubes containing 1M hydrochloric acid. The plasma
concentrations of the phosphate compound (prodrug) and its
hydroxylated homologue (drug) are determined simultaneously using a
method of liquid chromatography coupled with mass spectrometric
detection (TFC-LC-MS/MS). The limit of detection for both entities
is 0.5 ng/mL.
[0501] Lipid Formulation of the Drug from the Compounds of Examples
20 and 25
[0502] The drugs from Examples 20 and 25 are prepared in a mixture
of polyethylene glycol 300/ethanol/Phosal 50PG (30/10/60, v/v/v)
intended for administration by the p.o. route to SCID mice, under
the following conditions: [0503] Group 1: 3 mg/kg p.o. (gavage, 10
mL/kg), [0504] Group 2: 25 mg/kg p.o. (gavage, 10 mL/kg).
[0505] Blood samples are taken at the following points in time (3
animals for each point in time): 0.25 hr, 0.5 hr, 1 hr, 2 hrs, 6
hrs and 24 hrs after oral administration.
[0506] The blood thereby collected is centrifuged and the plasma is
transferred to tubes containing 1M hydrochloric acid. The plasma
concentrations of the drug are determined using a method of liquid
chromatography coupled with mass spectrometric detection
(TFC-LC-MS/MS). The limit of quantification is 0.5 ng/mL.
[0507] Non-compartmental pharmacokinetic analysis is carried out.
The mean results are shown in Tables 6, 7, 8 and 9 hereinbelow.
TABLE-US-00017 TABLE 6 Example 20 Compound Compounds measured
administered Example 20 Drug from Example 20 Example 20, C.sub.max
(ng/mL) = BLQ C.sub.max (ng/mL) = 56 3 mg/kg p.o. T.sub.max (h) =
ND T.sub.max (h) = 1.0 Aqueous AUC.sub.t (ng h/mL) = ND AUC.sub.t
(ng h/mL) = 51 formulation Example 20, C.sub.max (ng/mL) = 127
C.sub.max (ng/mL) = 3701 25 mg/kg p.o. T.sub.max (h) = 0.25
T.sub.max (h) = 1.0 Aqueous AUC.sub.t (ng h/mL) = 106 AUC.sub.t (ng
h/mL) = 8724 formulation ND: not determined BLQ: below the limit of
quantification
TABLE-US-00018 TABLE 7 Example 20 Compound measured Compound
administered Drug from Example 20 Drug from Example 20, 3 mg/kg
p.o. C.sub.max (ng/mL) = 39 Lipid formulation T.sub.max (h) = 1.0
AUC.sub.t (ng h/mL) = 55 Drug from Example 20, 25 mg/kg p.o.
C.sub.max (ng/mL) = 5524 Lipid formulation T.sub.max (h) = 2.0
AUC.sub.t (ng h/mL) = 10172
TABLE-US-00019 TABLE 8 Example 25 Compound Compounds measured
administered Example 25 Drug from Example 25 Example 25, C.sub.max
(ng/mL) = 17 C.sub.max (ng/mL) = 29 3 mg/kg p.o. T.sub.max (h) =
1.0 T.sub.max (h) = 1.0 Aqueous AUC.sub.t (ng h/mL) = 14 AUC.sub.t
(ng h/mL) = 31 formulation Example 25, C.sub.max (ng/mL) = 106
C.sub.max (ng/mL) = 2232 25 mg/kg p.o.T.sub.max (h) = 1.0 T.sub.max
(h) = 1.0 Aqueous AUC.sub.t (ng h/mL) = 114 AUC.sub.t (ng h/mL) =
3965 formulation
TABLE-US-00020 TABLE 9 Example 25 Compound measured Compound
administered Drug from Example 25 Drug from Example 25, 3 mg/kg
p.o. C.sub.max (ng/mL) = 33 Lipid formulation T.sub.max (h) = 1.0
AUC.sub.t (ng h/mL) = 37 Drug from Example 25, 25 mg/kg p.o.
C.sub.max (ng/mL) = 3004 Lipid formulation T.sub.max (h) = 1.0
AUC.sub.t (ng h/mL) = 5704
[0508] The results show that, whatever the dose (3 or 25 mg/kg),
the major part of the prodrugs of formula (I) is rapidly converted
in vivo into the corresponding drugs of formula (I') (see Tables 6,
7, 8 and 9). Plasma exposures of the prodrugs (C.sub.max, AUC) are
low in comparison to exposures of the corresponding drugs. The
results also show that the plasma concentrations of the drugs so
measured (after administration of the prodrugs) are equivalent to
those measured after direct administration of the drugs by the oral
route (see Tables 7 and 9).
EXAMPLE D
In Vivo Pharmacokinetic Profile of the Compounds of Formula
(I')
[0509] The pharmacokinetic profile of
N-(4-hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-
-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-
-1-indolizine carboxamide (also referred to as the drug from
Example 1) is also evaluated in a lipid and aqueous formulation in
the Wistar rat.
[0510] The drug from Example 1 is prepared in an aqueous suspension
in hydroxyethylcellulose 1% (w/v) in water and compared to a lipid
formulation composed of a mixture of anhydrous ethanol/polyethylene
glycol 400/Phosal 50PG (10/30/60, v/v/v). The two formulations are
administered by the oral route to male Wistar rats (3 rats per
formulation) at a dose of 100 mg/kg p.o. (gavage, 10 mL/kg).
[0511] Blood samples are taken at the following points in time from
each animal (3 animals/point in time): 0.25 hr, 0.5 hr, 0.75 hr, 1
hr, 2 hrs, 4 hrs, 8 hrs and 24 hrs after oral administration.
[0512] The plasma concentrations of the tested compound are
determined after extraction followed by liquid chromatography
coupled with mass spectrometric detection. The limit of
quantification is 0.1 ng/mL. The results are presented in the Table
hereinbelow:
TABLE-US-00021 TABLE 10 Compound measured Compound administered
Drug from Example 1 Drug from Example 1, 100 mg/kg p.o. C.sub.max
(ng/mL) = 816 Aqueous formulation AUC.sub.t (ng h/mL) = 3480 Drug
from Example 1, 100 mg/kg p.o. C.sub.max (ng/mL) = 5070 Lipid
formulation AUC.sub.t (ng h/mL) = 42900
[0513] The results show that the lipid formulation makes possible
much better plasma exposure of the drug from Example 1 than the
aqueous formulation.
EXAMPLE E
In Vitro Test on Human Caco-2 Cells
[0514] The cellular transport from A to B (Apical to Basolateral)
of the phosphate compounds of formula (I) and the compounds of
formula (I') (corresponding drugs) is studied in human Caco-2
cells. Each compound is deposited apically at 1 or 3 .mu.M (in
duplicate) and then incubated for 120 minutes.
[0515] Several samples are taken during the experiment; [0516]
apically: immediately after deposition (t=0) and at 120 minutes
[0517] basolaterally: at the end of the experiment (120
minutes)
[0518] The concentrations of the phosphate compound (prodrug)
and/or of its hydroxylated homologue (drug) are determined by
liquid chromatography coupled with mass spectrometric detection
(LC-MS/MS). The limit of quantification for both entities is 2
ng/mL.
[0519] The apparent permeability (P.sub.app) and the predicted
absorbed fraction (F.sub.abs) in humans are calculated for the
prodrug, for the drug after incubation of the prodrug and for the
drug after incubation of the drug (Hubatsch et al, Nat Protoc.
2007; 2(9), 2111-2119).
[0520] The experiment yield, which corresponds to the ratio (in
percent) of the total amount of compound found at the end of the
experiment versus that incubated, is also calculated.
[0521] The results have been collated in Table 11. They show that
the prodrugs of the compounds of formula (I) are markedly
decomposed in the course of the experiment (experiment yields of
<1.5%), thereby bringing about the formation of the associated
drugs in substantial amounts.
[0522] At the end, the predicted absorbed fraction in humans for
the drugs formed after incubation of the prodrugs is similar to
that obtained after incubation of the drugs.
TABLE-US-00022 TABLE 11 Compound administered Compounds measured
Example 1 Drug from Example 1 Example 1 P.sub.app (10.sup.-6 cm/s)
= 0.01 P.sub.app (10.sup.-6 cm/s) = 0.83 F.sub.abs (%) = ND
F.sub.abs (%) = 71 Yield (%) = 0 Yield (%) = 42 Drug from Example 1
P.sub.app (10.sup.-6 cm/s) = 0.65 F.sub.abs (%) = 67 Yield (%) = 37
Example 4 Drug from Example 4 Example 4 P.sub.app (10.sup.-6 cm/s)
= 0.33 P.sub.app (10.sup.-6 cm/s) = 0.43 F.sub.abs (%) = ND
F.sub.abs (%) = 69 Yield (%) = 1.3 Yield (%) = 38 Drug from Example
4 P.sub.app (10.sup.-6 cm/s) = 0.21 F.sub.abs (%) = 46 Yield (%) =
20 Example 5 Drug from Example 5 Example 5 P.sub.app (10.sup.-6
cm/s) = 0.26 P.sub.app (10.sup.-6 cm/s) = 2.3 F.sub.abs (%) = ND
F.sub.abs (%) = 86 Yield (%) = 1.2 Yield (%) = 78 Drug from Example
5 P.sub.app (10.sup.-6 cm/s) = 0.7 F.sub.abs (%) = 68 Yield (%) =
34 Example 20 Drug from Example 20 Example 20 P.sub.app (10.sup.-6
cm/s) = 0 P.sub.app (10.sup.-6 cm/s) = 0.16 F.sub.abs (%) = ND
F.sub.abs (%) = 16 Yield (%) = 0.94 Yield (%) = 100 Drug from
Example 20 P.sub.app (10.sup.-6 cm/s) = 0.29 F.sub.abs (%) = 25
Yield (%) = 91 Example 21 Drug from Example 21 Example 21 P.sub.app
(10.sup.-6 cm/s) = 0 P.sub.app (10.sup.-6 cm/s) = 0.21 F.sub.abs
(%) = ND F.sub.abs (%) = 19 Yield (%) = 0.83 Yield (%) = 100 Drug
from Example 21 P.sub.app (10.sup.-6 cm/s) = 0.27 F.sub.abs (%) =
24 Yield (%) = 82 Example 25 Drug from Example 25 Example 25
P.sub.app (10.sup.-6 cm/s) = 0 P.sub.app (10.sup.-6 cm/s) = 0.22
F.sub.abs (%) = ND F.sub.abs (%) = 20 Yield (%) = 33 Yield (%) = 48
Drug from Example 25 P.sub.app (10.sup.-6 cm/s) = 0.49 F.sub.abs
(%) = 40 Yield (%) = 100 ND: not determined
EXAMPLE F
Anti-Tumour Activity in Vivo
[0523] The anti-tumour activity of the compounds of the invention
is evaluated in a xenograft model of RS4;11 leukaemia cells.
[0524] 1.times.10.sup.7 RS4;11 cells are grafted sub-cutaneously
into immunosuppressed mice (SCID strain). 25 to 30 days after the
graft, when the tumour mass has reached about 150 mm.sup.3, the
mice are treated orally with the various compounds in two different
regimes (daily treatment for five days per week for two weeks, or
two treatments per week for two weeks). The tumour mass is measured
twice a week from the start of treatment.
[0525] The compounds of the invention have antitumour activity, via
the oral route, in the RS4;11 leukaemia model (acute lymphoblastic
leukaemia). The results obtained show that the compounds of the
invention are capable of inducing significant tumour
regression.
EXAMPLE G
Pharmaceutical Composition: Tablets
TABLE-US-00023 [0526] 1000 tablets containing a dose of 5 mg of a
compound 5 g selected from Examples 1 to 25 Wheat starch 20 g Maize
starch 20 g Lactose 30 g Magnesium stearate 2 g Silica 1 g
Hydroxypropylcellulose 2 g
* * * * *