U.S. patent application number 12/321607 was filed with the patent office on 2009-05-21 for 4-oxo-4,6,7,8-tetrahydro-pyrrolo[1,2-a]pyrazine-6-carboxamide compounds.
This patent application is currently assigned to LES LABORATOIRES SERVIER. Invention is credited to Alain Benoist, Guillaume De Nanteuil, Philippe Gloanec, Jean-Gilles Parmentier, Alain Rupin, Marie-Odile Vallez, Tony Verbeuren.
Application Number | 20090131668 12/321607 |
Document ID | / |
Family ID | 34896643 |
Filed Date | 2009-05-21 |
United States Patent
Application |
20090131668 |
Kind Code |
A1 |
De Nanteuil; Guillaume ; et
al. |
May 21, 2009 |
4-oxo-4,6,7,8-Tetrahydro-pyrrolo[1,2-a]pyrazine-6-carboxamide
compounds
Abstract
Compound of formula (I): ##STR00001## wherein: ##STR00002##
represents 1-oxidopyridyl substituted by the remainder of the
molecule in any one of the positions 2, 3 and 4, m and n, which may
be identical or different, each represent an integer of from 1 to
3, R.sub.1 represents hydrogen or alkyl, R.sub.2 and R.sub.3, which
may be identical or different, each represent an atom or group
selected from hydrogen, halogen, alkyl, hydroxy, acyloxy and
alkoxy, or, together with the carbon atom carrying them, form a
cycloalkane having from 3 to 6 carbon atoms, R.sub.4 and R.sub.5
each represent hydrogen, or are adjacent and, together with the
carbon atoms carrying them, form a benzo ring, Ar represents aryl
or heteroaryl. Medicinal products containing the same which are
useful as thrombin inhibitors.
Inventors: |
De Nanteuil; Guillaume;
(Suresnes, FR) ; Gloanec; Philippe; (Marly Le Roi,
FR) ; Parmentier; Jean-Gilles; (Issy Les Moulineaux,
FR) ; Benoist; Alain; (Franconville, FR) ;
Rupin; Alain; (Savonnieres, FR) ; Vallez;
Marie-Odile; (Montreuil, FR) ; Verbeuren; Tony;
(Vernouillet, FR) |
Correspondence
Address: |
THE FIRM OF HUESCHEN AND SAGE
SEVENTH FLOOR, KALAMAZOO BUILDING, 107 WEST MICHIGAN AVENUE
KALAMAZOO
MI
49007
US
|
Assignee: |
LES LABORATOIRES SERVIER
Suresnes Cedex
FR
|
Family ID: |
34896643 |
Appl. No.: |
12/321607 |
Filed: |
January 22, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
11079638 |
Mar 14, 2005 |
7498330 |
|
|
12321607 |
|
|
|
|
Current U.S.
Class: |
544/349 |
Current CPC
Class: |
A61P 9/00 20180101; A61P
7/02 20180101; A61P 9/02 20180101; A61P 7/00 20180101; A61P 9/14
20180101; A61P 9/10 20180101; A61P 43/00 20180101; C07D 487/04
20130101 |
Class at
Publication: |
544/349 |
International
Class: |
C07D 487/04 20060101
C07D487/04 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 19, 2004 |
FR |
04.02841 |
Claims
1. A method of treating a living animal body, including a human,
afflicted with a condition selected from stable and unstable
angina; disorders of thrombotic origin and/or giving rise to
thrombotic complications; complications of vascular and
cardiovascular diseases, including atherosclerosis, arteritis, and
venous disease; and disorders involving thrombin formation and/or
activity, comprising the step of administering to the living animal
body, including a human, a therapeutically effective amount of a
compound of formula (I): ##STR00017## wherein: ##STR00018##
represents 1-oxidopyridyl substituted by the remainder of the
molecule in any one of the positions 2, 3 and 4, m and n, which may
be identical or different, each represent an integer of from 1 to
3, R.sub.1 represents hydrogen or linear or branched
(C.sub.1-C.sub.6)alkyl, R.sub.2 and R.sub.3, which may be identical
or different, each represent an atom or group selected from
hydrogen, halogen, linear or branched (C.sub.1-C.sub.6)alkyl,
hydroxy, linear or branched (C.sub.1-C.sub.6)acyloxy and linear or
branched (C.sub.1-C.sub.6)alkoxy, or, together with the carbon atom
carrying them, form a cycloalkane having from 3 to 6 carbon atoms,
R.sub.4 and R.sub.5 each represent hydrogen, or are adjacent and,
together with the carbon atoms carrying them, form a benzo ring, Ar
represents aryl or heteroaryl, or an enantiomer or addition salt
thereof with a pharmaceutically acceptable acid, it being
understood that: "aryl" may be phenyl, biphenylyl or naphthyl, each
of those groups being optionally substituted by one or more
identical or different groups selected from: halogen, linear or
branched (C.sub.1-C.sub.6)alkyl optionally substituted by a
hydroxy, carboxy or carbamoyl group, the carbamoyl group being
itself optionally substituted by one or two linear or branched
(C.sub.1-C.sub.6)alkyl, linear or branched (C.sub.1-C.sub.6)alkoxy,
hydroxy, trihalo-(C.sub.1-C.sub.6)alkyl in which the alkyl moiety
may be linear or branched, amino optionally substituted by one or
two linear or branched (C.sub.1-C.sub.6)alkyl, carboxymethoxy, and
carbamoylmethoxy optionally N-substituted by one or two groups
selected from linear or branched (C.sub.1-C.sub.6)alkyl,
hydroxy-(C.sub.1-C.sub.6)alkyl in which the alkyl moiety may be
linear or branched, alkoxyalkyl in which the alkoxy and alkyl
moieties are each linear or branched C.sub.1-C.sub.6, and
pyridylalkyl in which the alkyl moiety is linear or branched
C.sub.1-C.sub.6, and "heteroaryl" may be a mono- or bi-cyclic
aromatic group having from 5 to 12 ring members and containing one,
two or three hetero atoms selected from oxygen, nitrogen and
sulphur, it being understood that the heteroaryl may be optionally
substituted by one or more identical or different groups selected
from: halogen, linear or branched (C.sub.1-C.sub.6)alkyl optionally
substituted by a hydroxy, carboxy or carbamoyl group, the carbamoyl
group being itself optionally substituted by one or two linear or
branched (C.sub.1-C.sub.6)alkyl, hydroxy, oxo, linear or branched
(C.sub.1-C.sub.6)alkoxy, trihalo-(C.sub.1-C.sub.6)alkyl in which
the alkyl moiety may be linear or branched, amino optionally
N-substituted by one or two linear or branched
(C.sub.1-C.sub.6)alkyl groups, carboxymethoxy, and carbamoylmethoxy
optionally N-substituted by one or two groups selected from linear
or branched (C.sub.1-C.sub.6)alkyl, hydroxy-(C.sub.1-C.sub.6)alkyl
in which the alkyl moiety may be linear or branched, alkoxyalkyl in
which the alkoxy and alkyl moieties are each linear or branched
C.sub.1-C.sub.6, and pyridylalkyl in which the alkyl moiety is
linear or branched C.sub.1-C.sub.6.
2. The method of claim 1, wherein the configuration of the
asymmetric centre at the alpha position with respect to the amide
of the compound of formula (I) is (S).
3. The method of claim 1, wherein m is 1.
4. The method of claim 1, wherein n is 1.
5. The method of claim 1, wherein R.sub.1 represents hydrogen.
6. The method of claim 1, wherein R.sub.2 represents hydrogen.
7. The method of claim 1, wherein R.sub.3 represents hydrogen.
8. The method of claim 1, wherein R.sub.4 and R.sub.5 each
represent hydrogen.
9. The method of claim 1, wherein R.sub.4 and R.sub.5 are adjacent
and, together with the carbon atoms carrying them, form a benzo
ring.
10. The method of claim 1 wherein Ar represents phenyl, thienyl or
pyridyl, each of those groups being unsubstituted or substituted by
one or more identical or different groups selected from: halogen,
linear or branched (C.sub.1-C.sub.6)alkyl optionally substituted by
hydroxy, carboxy or carbamoyl, the carbamoyl group being itself
optionally substituted by one or two linear or branched
(C.sub.1-C.sub.6)alkyl, linear or branched (C.sub.1-C.sub.6)alkoxy,
hydroxy, trihalo-(C.sub.1-C.sub.6)alkyl in which the alkyl moiety
may be linear or branched, amino optionally substituted by one or
two linear or branched (C.sub.1-C.sub.6)alkyl, carboxymethoxy, and
carbamoylmethoxy optionally N-substituted by one or two groups
selected from linear or branched (C.sub.1-C.sub.6)alkyl,
hydroxy-(C.sub.1-C.sub.6)alkyl in which the alkyl moiety may be
linear or branched, alkoxyalkyl in which the alkoxy and alkyl
moieties are each linear or branched C.sub.1-C.sub.6, and
pyridylalkyl in which the alkyl moiety is linear or branched
C.sub.1-C.sub.6.
11. The method of claim 10, wherein Ar represents phenyl
unsubstituted or substituted by one or more identical or different
halogen selected from fluorine and chlorine.
12. The method of claim 1, wherein the compound of formula (I) is
selected from:
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-fluorobenzyl)-4-o-
xo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide, and its
(6S) enantiomer,
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,6-difluorobenzyl)-
-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide, and
its (6S) enantiomer,
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-chlorobenzyl)-4-o-
xo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide, and its
(6S) enantiomer;
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,5-difluorobenzyl)-
-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide, and
its (6S) enantiomer,
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,3-difluorobenzyl)-
-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide, and
its (6S) enantiomer, and
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,3,6-trifluorobenz-
yl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide,
and its (6S) enantiomer.
13. The method of claim 1, wherein the compound of formula (I) is
administered in combination with one or more pharmaceutically
acceptable, inert, non-toxic carriers.
Description
[0001] The present invention relates to
4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
compounds.
BACKGROUND OF THE INVENTION
[0002] Thrombin is the key enzyme for coagulation and plays a
central role in the pathology of venous and arterial thromboses,
especially in view of its marked ability to cause
auto-amplification of the coagulation cascade (F. Toti et al.,
Sang, Thrombose, Vaisseaux 1992, 4, 483-494 and T. M. Reilly et
al., Blood Coagulation and Fibrinolysis 1992, 3, 513-517).
[0003] The direct and specific inhibition of thrombin is more
efficient and poses fewer risks of haemorrhage than treatment with
heparin. Direct inhibitors of thrombin currently exist but the
disadvantage of those peptide substances is that they are not
active when administered by the oral route.
DESCRIPTION OF THE PRIOR ART
[0004] Peptidomimetic compounds having an oral antithrombotic
activity have already been described in the literature. They
include, for example, the boronic acid compounds described in
Patent Specifications EP 293 881, EP 471 651, EP 615 978 and EP 792
883 and the compounds described in Patent Specifications WO 94
29336, WO 95 23609 and EP 1 069 132.
DETAILED DESCRIPTION OF THE INVENTION
[0005] The problem of the present invention was to obtain new
thrombin inhibitors active by the oral route that are
simultaneously well absorbed, potent, selective and reliable in
use. In that respect, it is important to have compounds that
present little risk of interaction with food or medicaments.
[0006] More specifically, the present invention relates to
compounds of formula (I):
##STR00003##
wherein:
##STR00004## [0007] represents a 1-oxidopyridyl group substituted
by the remainder of the molecule in any one of the positions 2, 3
and 4, [0008] m and n, which may be identical or different, each
represent an integer of from 1 to 3, [0009] R.sub.1 represents a
hydrogen atom or a linear or branched (C.sub.1-C.sub.6)alkyl group,
[0010] R.sub.2 and R.sub.3, which may be identical or different,
each represent an atom or group selected from the atoms hydrogen
and halogen and the groups linear or branched
(C.sub.1-C.sub.6)alkyl, hydroxy, linear or branched
(C.sub.1-C.sub.6)acyloxy and linear or branched
(C.sub.1-C.sub.6)alkoxy, or together form, with the carbon atom
carrying them, a cycloalkane having from 3 to 6 carbon atoms,
[0011] R.sub.4 and R.sub.5 each represent a hydrogen atom, or are
adjacent and together form, with the carbon atoms carrying them, a
benzo ring, [0012] Ar represents an aryl or heteroaryl group, to
their enantiomers, and to addition salts thereof with a
pharmaceutically acceptable acid.
[0013] 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, benzenesulphonic acid, camphoric acid, etc.
[0014] "Aryl group" is understood to mean phenyl, biphenylyl or
naphthyl, each of those groups being optionally substituted by one
or more identical or different groups selected from: [0015]
halogen, [0016] linear or branched (C.sub.1-C.sub.6)alkyl
optionally substituted by a hydroxy, carboxy or carbamoyl group,
the carbamoyl group being itself optionally substituted by one or
two linear or branched (C.sub.1-C.sub.6)alkyl groups, [0017] linear
or branched (C.sub.1-C.sub.6)alkoxy, [0018] hydroxy, [0019]
trihalo-(C.sub.1-C.sub.6)alkyl in which the alkyl moiety may be
linear or branched, [0020] amino optionally substituted by one or
two linear or branched (C.sub.1-C.sub.6)alkyl groups, [0021]
carboxymethoxy, [0022] and carbamoylmethoxy optionally
N-substituted by one or two groups selected from linear or branched
(C.sub.1-C.sub.6)alkyl, hydroxy-(C.sub.1-C.sub.6)alkyl in which the
alkyl moiety may be linear or branched, alkoxyalkyl group in which
the alkoxy and alkyl moieties are each linear or branched
C.sub.1-C.sub.6, and pyridylalkyl in which the alkyl moiety is
linear or branched C.sub.1-C.sub.6.
[0023] "Heteroaryl group" is understood to mean a mono- or
bi-cyclic aromatic group having from 5 to 12 ring members and
containing one, two or three hetero atoms selected from oxygen,
nitrogen and sulphur, it being understood that the heteroaryl may
be optionally substituted by one or more identical or different
groups selected from: [0024] halogen, [0025] linear or branched
(C.sub.1-C.sub.6)alkyl optionally substituted by a hydroxy, carboxy
or carbamoyl group, the carbamoyl group being itself optionally
substituted by one or two linear or branched (C.sub.1-C.sub.6)alkyl
groups, [0026] hydroxy, [0027] oxo, [0028] linear or branched
(C.sub.1-C.sub.6)alkoxy, [0029] trihalo-(C.sub.1-C.sub.6)alkyl in
which the alkyl moiety may be linear or branched, [0030] amino
optionally N-substituted by one or two linear or branched
(C.sub.1-C.sub.6)alkyl groups, [0031] carboxymethoxy, [0032] and
carbamoylmethoxy optionally N-substituted by one or two groups
selected from linear or branched (C.sub.1-C.sub.6)alkyl,
hydroxy-(C.sub.1-C.sub.6)alkyl in which the alkyl moiety may be
linear or branched, alkoxyalkyl in which the alkoxy and alkyl
moieties are each linear or branched C.sub.1-C.sub.6, and
pyridylalkyl in which the alkyl moiety is linear or branched
C.sub.1-C.sub.6.
[0033] Among the heteroaryl groups there may be mentioned, without
implying any limitation, the groups thienyl, pyridyl, furyl,
pyrrolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl,
isothiazolyl, pyrimidinyl, pyrazinyl, pyridazinyl, indolyl,
benzofuryl and quinolyl.
[0034] m is preferably 1.
[0035] n is preferably 1.
[0036] R.sub.1, R.sub.2 and R.sub.3 each preferably represent a
hydrogen atom.
[0037] An advantageous embodiment of the invention relates to
compounds of formula (I) wherein R.sub.4 and R.sub.5 each represent
a hydrogen atom.
[0038] Another advantageous embodiment of the invention relates to
compounds of formula (I) wherein R.sub.4 and R.sub.5 are adjacent
and together form, with the carbon atoms carrying them, a benzo
ring.
[0039] The group Ar is preferably a phenyl, thienyl or pyridyl
group, each of those groups being unsubstituted or substituted by
one or more identical or different groups selected from: [0040]
halogen, [0041] linear or branched (C.sub.1-C.sub.6)alkyl
optionally substituted by a hydroxy, carboxy or carbamoyl group,
the carbamoyl group being itself optionally substituted by one or
two linear or branched (C.sub.1-C.sub.6)alkyl groups, [0042] linear
or branched (C.sub.1-C.sub.6)alkoxy, [0043] hydroxy, [0044]
trihalo-(C.sub.1-C.sub.6)alkyl in which the alkyl moiety may be
linear or branched, [0045] amino optionally substituted by one or
two linear or branched (C.sub.1-C.sub.6)alkyl groups, [0046]
carboxymethoxy, [0047] and carbamoylmethoxy optionally
N-substituted by one or two groups selected from linear or branched
(C.sub.1-C.sub.6)alkyl, hydroxy-(C.sub.1-C.sub.6)alkyl in which the
alkyl moiety may be linear or branched, alkoxyalkyl in which the
alkoxy and alkyl moieties are each linear or branched
C.sub.1-C.sub.6, and pyridylalkyl in which the alkyl moiety is
linear or branched C.sub.1-C.sub.6.
[0048] More preferably, Ar represents a phenyl group unsubstituted
or substituted by one or more identical or different halogen atoms
selected from fluorine and chlorine.
[0049] Preferred compounds of formula (I) are [0050]
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-fluorobenzyl)-4-o-
xo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide, and its
(6S) enantiomer; [0051]
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,6-difluorobenzyl)-
-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide, and
its (6S) enantiomer; [0052]
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-chlorobenzyl)-4-o-
xo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide, and its
(6S) enantiomer; [0053]
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,5-difluorobenzyl)-
-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide, and
its (6S) enantiomer; [0054]
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino-}-N-(2,3-difluorobenzyl-
)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide, and
its (6S) enantiomer, [0055] and
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,3,6-trifluorobenz-
yl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide,
and its (6S) enantiomer.
[0056] The invention relates also to a process for the preparation
of compounds of formula (I) which is characterised in that a
compound of formula (II):
##STR00005##
wherein R.sub.2 and R.sub.3 are as defined for formula (I), P.sub.1
represents a protecting group for the amino function and Bn
represents a benzyl group is reduced, using a reducing agent, to
yield a compound of formula (III):
##STR00006##
wherein R.sub.2, R.sub.3, P.sub.1 and Bn are as defined
hereinbefore, the hydroxy function of which is converted to methoxy
and then to the cyano function by conventional reactions of organic
chemistry to yield, after deprotection of the amino function, a
compound of formula (IV):
##STR00007##
wherein R.sub.2, R.sub.3 and Bn are as defined hereinbefore, which
is reacted with oxalyl chloride to yield a compound of formula
(V):
##STR00008##
wherein R.sub.2, R.sub.3 and Bn are as defined hereinbefore, which
is subjected to catalytic hydrogenation to yield a compound of
formula (VI):
##STR00009##
wherein R.sub.2 and R.sub.3 are as defined hereinbefore, which is
esterified to form a compound of formula (VII):
##STR00010##
wherein R.sub.2 and R.sub.3 are as defined hereinbefore and P.sub.2
represents a linear or branched (C.sub.1-C.sub.6)alkyl group, which
is reacted with a brominating agent to yield a compound of formula
(VIII):
##STR00011##
wherein R.sub.2, R.sub.3 and P.sub.2 are as defined hereinbefore,
which is reacted with 2-mercaptopyridine to yield a compound of
formula (IX):
##STR00012##
wherein R.sub.2, R.sub.3 and P.sub.2 are as defined hereinbefore,
which is reacted with an N-oxide of formula (X):
##STR00013##
wherein m, R.sub.1, R.sub.4 and R.sub.5 are as defined for formula
(I), to yield a compound of formula (XI):
##STR00014##
wherein m, R.sub.1, R.sub.2, R.sub.3 R.sub.4, R.sub.5 and P.sub.2
are as defined hereinbefore, the acid function of which is
deprotected to yield a compound of formula (XII):
##STR00015##
wherein m, R.sub.1, R.sub.2, R.sub.3 R.sub.4 and R.sub.5 are as
defined hereinbefore, which is reacted with a compound of formula
(XIII):
##STR00016##
wherein n and Ar are as defined for formula (I), in the presence of
a coupling agent, to yield a compound of formula (I).
[0057] The addition salts of the compounds of formula (I) are
obtained by reaction of the compound with a pharmaceutically
acceptable acid.
[0058] The compounds of formula (I) have an asymmetric centre and
are therefore capable of existing in the form of a racemic mixture
or in optically active form.
[0059] The optically active compounds of formula (I) can be
obtained, for example, by using a corresponding optically active
compound of formula (II) as starting material or by separation of
the corresponding racemic mixture of formula (I), for example by
chiral HPLC chromatography.
[0060] Preferred compounds of formula (I) are those wherein the
configuration of the asymmetric centre in the alpha position with
respect to the amide is (S).
[0061] The compounds of formula (II) are obtained by benzylation of
the corresponding acids.
[0062] The compounds of the present invention have especially
valuable pharmacological properties.
[0063] They are potent inhibitors of thrombin which are active by
the oral route.
[0064] Those properties render them useful in the treatment of
stable or unstable angina, disorders of thrombotic origin and/or
giving rise to thrombotic complications, in the treatment or
prevention of myocardial infarction and venous or arterial
thromboses, and in the treatment of complications of vascular and
cardiovascular diseases such as atherosclerosis, arteritis, venous
disease, and in the treatment of any disorders involving thrombin
formation and/or activity.
[0065] They may also be used in therapeutic association with a
thrombolytic.
[0066] The invention relates also to pharmaceutical compositions
comprising as active ingredient a compound of formula (I) together
with one or more suitable inert, non-toxic excipients. Among the
pharmaceutical compositions according to the invention there may be
mentioned more especially those which are suitable for oral,
parenteral (intravenous or sub-cutaneous) or nasal administration,
tablets or dragees, sublingual tablets, gelatin capsules, lozenges,
suppositories, creams, ointments, dermal gels, injectable
preparations, drinkable suspensions, etc.
[0067] The useful dosage can be adapted according to the nature and
severity of the disorder, the route of administration and the age
and weight of the patient. The dosage varies from 1 to 500 mg per
day in one or more administrations.
[0068] The following Examples illustrate the invention.
[0069] The starting materials employed are known products or are
prepared according to known procedures.
[0070] The structures of the compounds described in the Examples
were determined according to customary spectrophotometric
techniques (infra-red NMR, mass spectrometry).
EXAMPLE 1
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-fluorobenzyl)-4-ox-
o-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
Step A: Benzyl N-tert-butoxycarbonyl-5-oxoprolinate
[0071] At 0.degree. C., 11 mmol of dimethylaminopyridine and 11
mmol of di-tert-butyl dicarbonate are added to 10 mmol of benzyl
5-oxoprolinate (the preparation process for which is described by
E. Campaigne et al. (J. Heterocycl. Chem. 1975, 12, 391)) in
solution in dichloromethane. After stirring for 24 hours at ambient
temperature, the reaction mixture is washed and then dried and
evaporated to yield the expected product in the form of a viscous
oil.
Step B: Benzyl N-tert-butoxycarbonyl-5-hydroxyprolinate
[0072] Under argon and at -78.degree. C., 18 mmol of a 1M solution
of diisobutylaluminium hydride in hexane are added to 10 mmol of
the compound obtained in the above Step in solution in
tetrahydrofuran. After stirring for 20 minutes at -78.degree. C., a
saturated aqueous solution of ammonium chloride is added, followed
by an aqueous 10% sodium carbonate solution. After stirring
overnight at ambient temperature, the reaction mixture is filtered
and the filtrate is evaporated and taken up in dichloromethane. The
organic phase is washed, dried and then evaporated. The residue is
purified by chromatography on silica gel, using a 95/5
dichloromethane/ethyl acetate mixture as eluant. The expected
product is obtained in the form of a yellow oil.
Step C: Benzyl N-tert-butoxycarbonyl-5-methoxyprolinate
[0073] A 0.1% solution of para-toluenesulphonic acid in anhydrous
methanol (88 ml) is added to 10 mmol of the compound obtained in
the above Step. After stirring for 1/2 hour, an aqueous 10% sodium
carbonate solution is added and the product is extracted with
dichloromethane. The expected product is obtained in the form of a
slightly yellow oil.
Step D: Benzyl 5-cyanoprolinate Hydrochloride
[0074] At .about.40.degree. C. and under argon, a 5% v/v solution
of tin tetrachloride in anhydrous dichloromethane (7.1 ml), and
then trimethylsilyl cyanide (20.6 mmol), are added to 10 mmol of
the compound obtained in the above Step. After stirring for 2 hours
at 40.degree. C., an aqueous 10% sodium carbonate solution is
added, the aqueous phase is extracted with dichloromethane, and the
organic phase is washed, dried and then evaporated. The residue
obtained is purified by chromatography on silica gel using a 95/5
dichloromethane/ethyl acetate mixture as eluant. The yellow oil
obtained is dissolved in ethyl acetate and then a stream of HCl gas
is passed through for 30 minutes at 0.degree. C. After stirring
overnight at ambient temperature, the precipitate formed is
filtered off, rinsed with ethyl acetate and dried in vacuo using a
dessicator.
Step E: Benzyl
1-chloro-3,4-dioxo-2,3,4,6,7,8-hexahydropyrrolo[1,2-a]pyrazine-6-carboxyl-
ate
[0075] At 0.degree. C., oxalyl chloride (144 ml) is added to 200 g
of the compound obtained in the above Step in solution in toluene.
The mixture is then brought to ambient temperature and stirred for
15 hours, and the solvent is subsequently evaporated off. The
residue obtained is purified by chromatography on silica, using a
9/1 dichloromethane/methanol mixture as eluant, to yield the
expected product.
Step F:
3,4-Dioxo-2,3,4,6,7,8-hexahydropyrrolo[1,2-a]pyrazine-6-carboxylic
Acid
[0076] 3 g of the compound obtained in the above Step are dissolved
in 50 ml of ethanol and then 1.43 ml of triethylamine are added,
followed by 0.5 g of palladium-on-carbon. The mixture is then
placed under a hydrogen atmosphere at ambient temperature and
atmospheric pressure for 5 hours. After removal of the catalyst by
filtration, the solvent is evaporated off to yield the expected
product.
[0077] Step G: Ethyl
3,4-dioxo-2,3,4,6,7,8-hexahydropyrrolo[1,2-a]pyrazine-6-carboxylate
[0078] 11.83 ml of trimethylsilyl chloride are added dropwise, at
0.degree. C., to a suspension of 1.83 g of the compound obtained in
the above Step in 20 ml of anhydrous ethanol. The reaction mixture
is then stirred at ambient temperature for 15 hours. The solvent is
evaporated off and the residue is taken up in dichloromethane. The
organic phase is washed, dried, filtered and evaporated, and then
the crude product is purified by chromatography on silica (eluant:
dichloromethane/ethanol 95/5) to yield the expected product.
Step H: Ethyl
3-bromo-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxylate
[0079] 34.7 g of dibasic sodium phosphate (Na.sub.2HPO.sub.4) in 90
ml of dichloroethane, and then 100 g of phosphorus oxybromide
(POBr.sub.3) in 340 ml of dichloroethane, are added to 73 g of the
compound obtained in the above Step suspended in 400 ml of
dichloroethane. The reaction mixture is then heated at 50.degree.
C. for 16 hours, subsequently cooled to 0.degree. C. using a bath
of water and ice, and 650 ml of a 10% sodium carbonate solution are
added.
[0080] The organic phase is washed with water, then the aqueous
phases are combined and extracted with isopropyl acetate. The
combined organic phases are concentrated to 160 ml, and then 160 ml
of isopropyl acetate are added and the mixture is again
concentrated to 160 ml. The mixture is subsequently placed in a
bath at 50.degree. C. and 250 ml of n-heptane are added in the
course of 1 hours. After a further hour at 50.degree. C., the
precipitate obtained is filtered off, rinsed with a 1/3 mixture of
isopropyl acetate and n-heptane, and dried to yield the expected
product.
Step I: Ethyl
4-oxo-3-(2-pyridylthio)-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carbox-
ylate
[0081] 34.3 g of 2-mercaptopyridine are added in three batches, at
20 minute intervals, to 80.5 g of the compound obtained in the
above Step in solution in 410 ml of acetonitrile. The addition is
exothermic and the temperature of the reaction mixture rises to
35.degree. C.
[0082] The mixture becomes heterogeneous. After stirring for 1 hour
45 minutes, the solvent is evaporated off and the residue is taken
up in ethyl acetate and in water. The organic phase is washed,
dried, filtered and evaporated to dryness. The residue obtained is
purified on a chromatography column (dichloromethane/ethanol 98/2
then 95/5) to yield the expected product in the form of an orange
oil which slowly crystallises.
Step J: Ethyl
3-{[2,2-difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-4-oxo-4,6,7,8-tetrahyd-
ropyrrolo[1,2-a]pyrazine-6-carboxylate
[0083] 2.78 g of 2,2-difluoro-2-(1-oxido-2-pyridyl)ethanamine, and
then 1.5 g zinc chloride, are added to 4.6 g of the compound
obtained in the above Step suspended in 30 ml of acetonitrile. The
mixture is then heated at reflux for 15 hours. The reaction mixture
becomes clear. The solvent is then evaporated off and the residue
is taken up in dichloromethane. The organic phase is washed, dried,
filtered and evaporated, and the residue obtained is purified by
chromatography on silica (eluant: dichloromethane/isopropanol 9/1)
to yield the expected product.
Step K:
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-4-oxo-4,6,7,8-t-
etrahydropyrrolo[1,2-a]pyrazine-6-carboxylic Acid
[0084] 2 equivalents of 1N sodium hydroxide solution are added to 9
g of the compound obtained in the above Step in solution in 100 ml
of dioxane and 20 ml of water. After 24 hours at ambient
temperature, acidification is carried out using 2 equivalents of 1N
hydrochloric acid, and then the reaction mixture is evaporated. The
residue is taken up twice in 30 ml of toluene and dried, yielding
the expected product in the form of a white solid.
Step L:
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-fluorobenz-
yl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0085] 3 g of the compound obtained in the above Step are reacted
with 1.18 g of 2-fluorobenzylamine in the presence of 3.57 g of
1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium
3-oxide hexafluorophosphate (HATU) and 1.64 ml of
diisopropylethylamine in 60 ml of dimethylformamide.
[0086] After stirring for 15 hours at ambient temperature, the
solvent is evaporated off and the residue obtained is taken up in
ethyl acetate and in water. The organic phase is washed, dried and
then evaporated, and the residue is purified by chromatography on
silica to yield the expected product in the form of a racemic
mixture.
Elemental Microanalysis:
TABLE-US-00001 [0087] % C % H % N Calculated: 57.52 4.39 15.24
Found: 57.63 4.18 15.05
EXAMPLE 2
(6R)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-fluorobenzyl)-
-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0088] The racemic mixture of Example 1 is separated by chiral
phase preparative HPLC chromatography (Chiralpak AD column, eluant:
acetonitrile/isopropanol/diethylamine 500/500/1).
[0089] The expected product is the first of the enantiomers so
obtained.
EXAMPLE 3
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-fluorobenzyl)-
-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0090] The racemic mixture of Example 1 is separated by chiral
phase preparative HPLC chromatography (Chiralpak AD column, eluant:
acetonitrile/isopropanol/diethylamine 500/500/1).
[0091] The expected product is the second of the enantiomers so
obtained.
[0092] Index of rotation: .alpha..sub.D=-116.07.degree. (methanol,
20.degree. C., c=1.4)
EXAMPLE 4
(6R)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-{2-[2-(ethylamin-
o)-2-oxoethoxy]benzyl}-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-ca-
rboxamide
[0093] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 2-[2-(aminomethyl)phenoxy]-N-ethylacetamide in Step L,
followed by separation of the racemic mixture so obtained by
preparative chiral HPLC chromatography (Chiralpak AD column, eluant
acetonitrile/isopropanol/diethylamine 500/500/1).
[0094] The expected product is the first of the enantiomers so
obtained.
Elemental Microanalysis:
TABLE-US-00002 [0095] % C % H % N Calculated: 57.56 5.20 15.49
Found: 58.11 5.15 15.55
[0096] Index of rotation: .alpha..sub.D=+93.1.degree. (methanol,
20.degree. C., c=0.7)
EXAMPLE 5
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-{2-[2-(ethylamin-
o)-2-oxoethoxy]benzyl}-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-ca-
rboxamide
[0097] The expected product is the second of the enantiomers
separated in Example 4.
[0098] Index of rotation: .alpha..sub.D=-98.8.degree. (methanol,
20.degree. C., c=0.8)
EXAMPLE 6
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,4-difluorobenzyl)--
4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0099] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 2,4-difluorobenzylamine in Step L.
[0100] Mass spectrometry ESI (acetonitrile/water):
[M+H].sup.+=478.15.
EXAMPLE 7
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,6-difluorobenzyl)--
4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0101] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 2,6-difluorobenzylamine in Step L.
[0102] Melting point: 227-228.degree. C.
Elemental Microanalysis:
TABLE-US-00003 [0103] % C % H % N Calculated: 55.35 4.01 14.67
Found: 55.04 4.04 14.28
EXAMPLE 8
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-chlorobenzyl)-4-ox-
o-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0104] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 2-chlorobenzylamine in Step L.
Elemental Microanalysis:
TABLE-US-00004 [0105] % C % H % N % Cl Calculated: 55.53 4.24 14.72
7.45 Found: 55.32 4.33 14.33 7.73
EXAMPLE 9
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(3,4-difluorobenzyl)--
4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0106] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 3,4-difluorobenzylamine in Step L.
[0107] Melting point: 204.degree. C.
Elemental Microanalysis:
TABLE-US-00005 [0108] % C % H % N Calculated: 55.35 4.01 14.67
Found: 55.07 3.92 14.40
EXAMPLE 10
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,5-difluorobenzyl)--
4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0109] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 2,5-difluorobenzylamine in Step L.
[0110] Melting point: 189.degree. C.
Elemental Microanalysis:
TABLE-US-00006 [0111] % C % H % N Calculated: 55.35 4.01 14.67
Found: 55.95 4.41 14.13
EXAMPLE 11
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,6-dichlorobenzyl)--
4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0112] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 2,6-dichlorobenzylamine in Step L.
[0113] Melting point: 122.degree. C.
Elemental Microanalysis:
TABLE-US-00007 [0114] % C % H % N % Cl Calculated: 51.78 3.75 13.72
13.89 Found: 52.23 3.73 13.66 14.06
EXAMPLE 12
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-chloro-6-fluoroben-
zyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0115] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 2-chloro-6-fluorobenzylamine in Step L.
[0116] Melting point: 221.degree. C.
Elemental Microanalysis:
TABLE-US-00008 [0117] % C % H % N % Cl Calculated: 53.50 3.88 14.18
7.18 Found: 53.93 3.97 14.06 7.28
EXAMPLE 13
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,3-difluorobenzyl)--
4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0118] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 2,3-difluorobenzylamine in Step L.
[0119] Melting point: 142.degree. C.
Elemental Microanalysis:
TABLE-US-00009 [0120] % C % H % N Calculated: 55.35 4.01 14.67
Found: 55.34 4.26 14.42
EXAMPLE 14
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(3,5-difluorobenzyl)--
4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0121] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 3,5-difluorobenzylamine in Step L.
[0122] Melting point: 218-219.degree. C.
Elemental Microanalysis:
TABLE-US-00010 [0123] % C % H % N Calculated: 55.35 4.01 14.67
Found: 54.72 3.82 14.40
EXAMPLE 15
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-4-oxo-N-[2-(2-oxo-2-{[2-
-(2-pyridyl)ethyl]amino}ethoxy)benzyl]-4,6,7,8-tetrahydropyrrolo[1,2-a]pyr-
azine-6-carboxamide Hydrochloride
Step A:
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-4-oxo-N-[2-(2-o-
xo-2-{[2-(2-pyridyl)ethyl]amino}ethoxy)benzyl]-4,6,7,8-tetrahydropyrrolo[1-
,2-a]pyrazine-6-carboxamide
[0124] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 2-[2-(aminomethyl)phenoxy]-N-[2-(2-pyridyl)ethyl]acetamide in
Step L.
Step B:
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-4-oxo-N-[2-(2-o-
xo-2-{[2-(2-pyridyl)ethyl]amino}ethoxy)benzyl]-4,6,7,8-tetrahydropyrrolo[1-
,2-a]pyrazine-6-carboxamide Hydrochloride
[0125] The expected product is obtained by acidification of the
compound obtained in the above Step using hydrochloric acid.
Elemental Microanalysis:
TABLE-US-00011 [0126] % C % H % N % Cl Calculated: 56.75 4.92 14.94
5.40 Found: 57.22 4.85 14.87 5.82
EXAMPLE 16
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,6-difluoroben-
zyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0127] The expected product is obtained by separation of the
racemic mixture of Example 7 on a chiral HPLC column.
[0128] Index of rotation: .alpha..sub.D=-118.46.degree. (methanol,
20.degree. C., c=0.95)
EXAMPLE 17
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,5-difluoroben-
zyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0129] The expected product is obtained by separation of the
racemic mixture of Example 10 on a chiral HPLC column.
[0130] Index of rotation: .alpha..sub.D=-89.65.degree. (methanol,
20.degree. C., c=0.57)
EXAMPLE 18
(6S-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-chloro-6-fluor-
obenzyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0131] The expected product is obtained by separation of the
racemic mixture of Example 12 on a chiral HPLC column.
[0132] Index of rotation: .alpha..sub.D=-101.49.degree. (methanol,
20.degree. C., c=1.3)
EXAMPLE 19
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,3-difluoroben-
zyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0133] The expected product is obtained by separation of the
racemic mixture of Example 13 on a chiral HPLC column.
[0134] Index of rotation: .alpha..sub.D=-102.06.degree. (methanol,
20.degree. C., c=0.8)
EXAMPLE 20
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-chlorobenzyl)-
-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0135] The expected product is obtained by separation of the
racemic mixture of Example 8 on a chiral HPLC column.
[0136] Index of rotation: .alpha..sub.D=-105.65.degree. (methanol,
20.degree. C., c=0.85)
EXAMPLE 21
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,3,6-trifluoro-
benzyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
Step A:
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,3,6-triflu-
orobenzyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0137] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 2,3,6-trifluorobenzylamine in Step L.
Step B:
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2,3,6-t-
rifluorobenzyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxami-
de
[0138] The expected product is obtained by separation of the
racemic mixture obtained in the above Step on a chiral HPLC
column.
[0139] Index of rotation: .alpha..sub.D=-118.25 (methanol,
20.degree. C., c=1)
EXAMPLE 22
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-benzyl-4-oxo-4,6-
,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
Hydrochloride
Step A:
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-benzyl-4-oxo--
4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0140] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with benzylamine in Step L.
Step B:
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-benzyl-4-
-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0141] The expected product is obtained by separation of the
racemic mixture obtained in the above Step on a chiral HPLC
column.
Step C:
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-benzyl-4-
-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
Hydrochloride
[0142] The expected product is obtained by acidification of the
compound obtained in the above Step using hydrochloric acid.
Elemental Microanalysis:
TABLE-US-00012 [0143] % C % H % N % Cl Calculated: 55.29 4.64 14.65
7.42 Found: 54.84 4.81 14.16 7.16
EXAMPLE 23
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(3-thienylmethyl-
)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
Hydrochloride
[0144] The expected product is obtained according to the procedure
described in Example 22, with the replacement of benzylamine with
3-thienylmethylamine in Step A.
Elemental Microanalysis:
TABLE-US-00013 [0145] % C % H % N % Cl % S Calculated: 49.64 4.17
14.47 7.33 6.63 Found: 49.09 4.17 14.14 8.16 6.59
EXAMPLE 24
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-N-(2-thienylmethyl-
)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
Hydrochloride
[0146] The expected product is obtained according to the procedure
described in Example 22, with the replacement of benzylamine with
2-thienylmethylamine in Step A.
Elemental Microanalysis:
TABLE-US-00014 [0147] % C % H % N % Cl % S Calculated: 49.64 4.17
14.47 7.33 6.63 Found: 50.61 4.12 14.35 7.98 6.65
EXAMPLE 25
(6S)-3-{[2,2-Difluoro-2-(1-oxido-4-pyridyl)ethyl]amino}-N-(2-fluorobenzyl)-
-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
Step A:
3-{[2,2-Difluoro-2-(1-oxido-4-pyridyl)ethyl]amino}-N-(2-fluorobenz-
yl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0148] The expected product is obtained according to the procedure
described in Example 1, with the replacement of
2,2-difluoro-2-(1-oxido-2-pyridyl)ethanamine with
2,2-difluoro-2-(1-oxido-4-pyridyl)ethanamine in Step J.
Step B:
(6S)-3-{[2,2-Difluoro-2-(1-oxido-4-pyridyl)ethyl]amino}-N-(2-fluor-
obenzyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0149] The expected product is obtained by separation of the
racemic mixture obtained in the above Step on a chiral HPLC
column.
Elemental Microanalysis:
TABLE-US-00015 [0150] % C % H % N Calculated: 57.52 4.39 15.24
Found: 57.73 4.61 15.05
EXAMPLE 26
(6S)-3-{[2,2-Difluoro-2-(1-oxido-3-pyridyl)ethyl]amino}-N-(2-fluorobenzyl)-
-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0151] The expected product is obtained according to the procedure
of Example 25, with the replacement of
2,2-difluoro-2-(1-oxido-4-pyridyl)ethanamine with
2,2-difluoro-2-(1-oxido-3-pyridyl)ethanamine in Step A.
Elemental Microanalysis:
TABLE-US-00016 [0152] % C % H % N Calculated: 57.52 4.39 15.24
Found: 57.34 4.49 14.99
EXAMPLE 27
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-8,8-dimethyl-N-(2-fluor-
obenzyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0153] The expected product is obtained according to the procedure
of Example 1, with the replacement of benzyl 5-oxoprolinate with
benzyl 4,4-dimethyl-5-oxo-2-pyrrolidine-carboxylate in Step A.
EXAMPLE 28
(6R)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-8,8-dimethyl-N-(2--
fluorobenzyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0154] The racemic mixture of Example 27 is separated by chiral
phase preparative HPLC chromatography.
[0155] The expected product is the first of the enantiomers so
obtained.
Elemental Microanalysis:
TABLE-US-00017 [0156] % C % H % N Calculated: 59.13 4.96 14.37
Found: 58.88 4.99 14.08
EXAMPLE 29
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-8,8-dimethyl-N-(2--
fluorobenzyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0157] The racemic mixture of Example 27 is separated by chiral
phase preparative HPLC chromatography.
[0158] The expected product is the second of the enantiomers so
obtained.
Elemental Microanalysis:
TABLE-US-00018 [0159] % C % H % N Calculated: 59.13 4.96 14.37
Found: 58.92 4.93 14.14
EXAMPLE 30
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-4-oxo-N-[(5-oxo-4,-
5-dihydro-1H-1,2,4-triazol-3-yl)methyl]-4,6,7,8-tetrahydropyrrolo[1,2-a]py-
razine-6-carboxamide
Step A:
3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-4-oxo-N-[(5-oxo-
-4,5-dihydro-1H-1,2,4-triazol-3-yl)methyl]-4,6,7,8-tetrahydropyrrolo[1,2-a-
]pyrazine-6-carboxamide
[0160] The expected product is obtained according to the procedure
described in Example 1, with the replacement of 2-fluorobenzylamine
with 5-(aminomethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one in Step
L.
Step B:
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-pyridyl)ethyl]amino}-4-oxo-N-[(-
5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)methyl]-4,6,7,8-tetrahydropyrrolo[-
1,2-a]pyrazine-6-carboxamide
[0161] The expected product is obtained by separation of the
racemic mixture obtained in the above Step on a chiral HPLC
column.
Elemental Microanalysis:
TABLE-US-00019 [0162] % C % H % N Calculated: 48.22 4.05 24.99
Found: 48.61 4.34 24.56
EXAMPLE 31
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-quinolinyl)ethyl]amino}-N-(2-fluorobenz-
yl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
Step A:
3-{[2,2-Difluoro-2-(1-oxido-2-quinolinyl)ethyl]amino}-N-(2-fluorob-
enzyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0163] The expected product is obtained according to the procedure
described in Example 1, with the replacement of
2,2-difluoro-2-(1-oxido-2-pyridyl)ethanamine with
2,2-difluoro-2-(1-oxido-2-quinolinyl)ethanamine in Step J.
Step B:
(6S)-3-{[2,2-Difluoro-2-(1-oxido-2-quinolinyl)ethyl]amino}-N-(2-fl-
uorobenzyl)-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]pyrazine-6-carboxamide
[0164] The expected product is obtained by separation of the
racemic mixture obtained in the above Step on a chiral HPLC
column.
Elemental Microanalysis:
TABLE-US-00020 [0165] % C % H % N Calculated: 61.29 4.35 13.75
Found: 61.65 4.51 13.50
[0166] Pharmacological Study of the Compounds of the Invention
EXAMPLE 32
Inhibition of Thrombin and of Fibrinolysis Serine Proteases
[0167] For in vitro evaluation of the inhibitory activity of the
products of the invention on human thrombin (Sigma, specific
activity 3230 UNIH/mg), purified human fibrinogen (4 mM, Stago)
(Fg) was added to a given amount of thrombin (0.7 nM) that had
previously been incubated with or without the inhibitor to be
tested (20.degree. C., 10 minutes).
[0168] Inhibitors, enzymes and substrates are diluted in the same
buffer (0.01 mM phosphate buffer, pH 7.4, containing 0.12 M sodium
chloride and 0.05% bovine serum albumin) and then distributed on a
polystyrene microtitre plate in a volume of 50 .mu.l.
[0169] The fibrin formed by the thrombin is measured using a
spectrophotometer at 405 nm after from 10 to 15 minutes' reaction
at 20.degree. C.
[0170] The table below gives in nM the concentration of the
compounds that inhibits 50% of the enzymatic activity (IC.sub.50)
of the thrombin compared with the control without product. The
results obtained show that the compounds of the invention are
potent inhibitors of the activity of human thrombin on human
fibrinogen.
TABLE-US-00021 TABLE Example IC.sub.50 (nM) 1 28 3 16 8 38 15 1.4
21 11 24 60 31 8.4
EXAMPLE 33
Pharmaceutical Composition
[0171] Formulation for the preparation of 1000 tablets each
containing a dose of 10 mg:
TABLE-US-00022 compound of Example 3 10 g hydroxypropyl cellulose 2
g wheat starch 10 g lactose 100 g magnesium stearate 3 g talc 3
g
* * * * *