U.S. patent application number 11/742048 was filed with the patent office on 2007-08-30 for use of 2-amino-thiazoline derivatives as inhibitors of inducible no-synthase.
This patent application is currently assigned to AVENTIS PHARMA S.A.. Invention is credited to Antony BIGOT, Jean-Christophe CARRY, Serge MIGNANI.
Application Number | 20070203337 11/742048 |
Document ID | / |
Family ID | 26213260 |
Filed Date | 2007-08-30 |
United States Patent
Application |
20070203337 |
Kind Code |
A1 |
BIGOT; Antony ; et
al. |
August 30, 2007 |
USE OF 2-AMINO-THIAZOLINE DERIVATIVES AS INHIBITORS OF INDUCIBLE
NO-SYNTHASE
Abstract
The present invention relates to a process for preparing
2-amino-thiazoline derivatives of formula (II): ##STR1## in which
either Y is a methylene (CH.sub.2) and X is chosen from the
following groups: O, NH, (C1-C4) N-Alkyl, N-Bn, N-Ph, N-(2-Py),
N-(3-Py), N-(4-Py), N-2-pyrimidyl, N-5-pyrimidyl, S, SO, SO.sub.2,
CH.sub.2 or CHPh; or Y is a carbonyl (C.dbd.O) and X is chosen from
the following groups: NH, N-Ph, N-(2-Py), N-(3-Py), N-(4-Py),
N-2-pyrimidyl or N-5-pyrimidyl.
Inventors: |
BIGOT; Antony; (Massy,
FR) ; CARRY; Jean-Christophe; (Saint Maur Des Fosses,
FR) ; MIGNANI; Serge; (Chatenay-Malabry, FR) |
Correspondence
Address: |
ROSS J. OEHLER;SANOFI-AVENTIS U.S. LLC
1041 ROUTE 202-206
MAIL CODE: D303A
BRIDGEWATER
NJ
08807
US
|
Assignee: |
AVENTIS PHARMA S.A.
20 Avenue Raymond Aron
Antony
FR
92160
|
Family ID: |
26213260 |
Appl. No.: |
11/742048 |
Filed: |
April 30, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11200713 |
Aug 10, 2005 |
7227022 |
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11742048 |
Apr 30, 2007 |
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10764853 |
Jan 26, 2004 |
6953796 |
|
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11200713 |
Aug 10, 2005 |
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10291084 |
Nov 8, 2002 |
6699867 |
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10764853 |
Jan 26, 2004 |
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60352797 |
Jan 30, 2002 |
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Current U.S.
Class: |
544/3 ; 540/1;
544/399 |
Current CPC
Class: |
C07D 417/14 20130101;
A61P 43/00 20180101; A61P 1/00 20180101; A61P 25/08 20180101; A61P
25/16 20180101; A61P 25/24 20180101; C07D 295/13 20130101; C07D
417/06 20130101; C07D 277/18 20130101; C07D 295/15 20130101; A61P
1/04 20180101; A61P 25/00 20180101; A61P 9/10 20180101; A61P 25/06
20180101; A61P 31/04 20180101; A61P 27/02 20180101; A61P 17/00
20180101; C07D 417/12 20130101; A61P 25/14 20180101; A61P 29/00
20180101; A61P 19/02 20180101; A61P 3/10 20180101; A61P 25/28
20180101; A61P 17/06 20180101; A61P 13/12 20180101; A61P 25/22
20180101; A61P 11/06 20180101; C07D 241/04 20130101; A61P 35/00
20180101; A61P 21/04 20180101 |
Class at
Publication: |
544/003 ;
540/001; 544/399 |
International
Class: |
C07D 241/04 20060101
C07D241/04; C07D 279/12 20060101 C07D279/12 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 9, 2001 |
FR |
0114510 |
Claims
1) An intermediate compound for the preparation of
2-amino-thiazoline derivatives, said intermediate being selected
from the compounds of formulas (IIa), (IIb), and (IIc), ##STR12##
wherein: Ra is a protecting group of the amine function and Rb is a
protecting group of the acid function, and in which either Y is a
methylene (CH.sub.2) and X is selected from O, NH, N--(C1-C4)alkyl,
N-Bn, N-Ph, N-(2-Py), N-(3-Py), N-(4-Py), N-2-pyrimidyl,
N-5-pyrimidyl, S, SO, SO.sub.2, CH.sub.2, and CHPh; or Y is a
carbonyl (C.dbd.O) and X is selected from: NH, N-Ph, N-(2-Py),
N-(-'-Py), N-(4-Py), N-2-pyrimidyl and N-5-pyrimidyl; said
(C1-C4)alkyl radicals containing 1 to 4 carbons in a straight or
branched chain, and said abbreviations Bn, Py and Ph signifying,
respectively, benzyl, pyridyl and phenyl excluding, however, the
following compounds:
N-[2-hydroxy-1-(4-phenyl-piperazin-1-ylmethyl)ethyl]thiobenzamide,
methyl 2-(N-CBZ-amino)-3-morpholino-propionate,
boc-DL-.beta.-(4-morpholinyl)alanine methyl ester, methyl
2-acetamido-3-piperidinopropanoate, methyl
2-acetamido-3-morpholinopropanoate, ethyl
2-(N-benzyl-amino)-3-piperidinopropanoate, and ethyl
2-(N-benzyl-amino)-3-morpholinopropanoate.
2) An intermediate compound according to claim 1, selected from the
group consisting of:
N-(tert-Butyl)-N'-[2-hydroxy-1-(4-methyl-piperazin-1-ylmethyl)ethyl]-thio-
urea;
N-[2-Hydroxy-1-(4-methyl-piperazin-1-ylmethyl)ethyl]acetamide; and
methyl 2-(Acetylamino)-3-(4-methyl-piperazin-1-yl)propanoate.
Description
[0001] This application is a Divisional of U.S. application Ser.
No. 11/200,713, filed Aug. 10, 2005, now allowed, which is a
Divisional of U.S. application Ser. No. 10/764,853, filed Jan. 26,
2004, now U.S. Pat. No. 6,953,796, issued Oct. 11, 2005, which is a
Continuation of U.S. application Ser. No. 10/291,084, filed Nov. 8,
2002, now U.S. Pat. No. 6,699,867, issued Mar. 2, 2004, which
claims the benefit of U.S. Provisional Application No. 60/352,797,
filed Jan. 30, 2002, and benefit of priority of French Patent
Application No. 01/14,510, filed Nov. 9, 2001; all of which are
incorporated herein by reference in their entirety.
[0002] The present invention relates to the production of
2-amino-thiazoline derivatives of formula (I): ##STR2## or
pharmaceutically acceptable salts thereof, which are useful as
inhibitors of inducible NO-synthase.
[0003] The subject of the invention is the use of
2-amino-thiazoline derivatives of formula (I) and pharmaceutically
acceptable salts thereof for the preparation of pharmaceutical
compositions intended for preventing and treating diseases in which
an abnormal production of nitric oxide (NO) by induction of
inducible NO-synthase (NOS-2 or iNOS) is involved, the
pharmaceutical compositions containing the novel 2-amino-thiazoline
derivatives and pharmaceutically acceptable salts thereof and the
novel derivatives of 2-amino-thiazoline and pharmaceutically
acceptable salts thereof.
[0004] Nitric oxide (NO) is a diffusible radical involved in many
physiological and pathological processes. It is synthesized by
oxidation of L-Arginine, a reaction catalyzed by a family of
enzymes known as nitric oxide synthases or NO-Synthase (NOS),
referenced in the international enzyme nomenclature under the
number E.C. 1.14.13.39.
[0005] Three NOS isoforms, two of which are constitutive and one
inducible, are known: [0006] a neuronal NOS (NOS-1 or nNOS) was
originally isolated and cloned from nerve tissue in which it is a
constitutive enzyme. The NOS-1 produces NO in response to various
physiological stimuli such as the activation of membrane receptors
according to a mechanism dependent on calcium and on calmodulin.
[0007] an inducible NOS (NOS-2 or iNOS) can be induced in response
to immunological stimuli such as, for example, cytokines or
bacterial antigens in various cells such as, for example,
macrophages, endothelial cells, hepatocytes, glial cells, as well
as many other types of cells. This isoform activity is not
regulated by calcium. Consequently, once induced, it produces a
large amount of NO over prolonged periods. [0008] an endothelial
NOS (NOS-3 or eNOS) is constitutive and calcium/calmodulin
dependent. It was originally identified in vascular endothelium
cells, in which it generates NO in response to physiological
stimuli such as the activation of membrane receptors.
[0009] The NO produced by the neuronal and endothelial constitutive
isoforms (NOS-1 and NOS-3) is generally involved in intercellular
signaling functions. For example, the endothelial cells which line
the inner wall of blood vessels induce the relaxation of the
underlying smooth muscular cells via the production de NO. It thus
contributes towards regulating the arterial pressure.
[0010] The NO produced in large amount by the inducible isoform
NOS-2 is, inter alia, involved in the pathological phenomena
associated with acute and chronic inflammatory processes in a large
variety of tissues and organs.
[0011] An excessive production of NO by induction of NOS-2 thus
plays a part in degenerative pathologies of the nervous system such
as, for example, multiple sclerosis, focal or global cerebral
ischemia, cerebral or spinal trauma, Parkinson's disease,
Huntington's disease, Alzheimer's disease, amyotrophic lateral
sclerosis, migraine, depression, schizophrenia, anxiety, epilepsy.
Similarly, aside the central nervous system, the induction of NOS-2
is involved in many pathologies with inflammatory components such
as, for example, diabetes, atherosclerosis, myocarditis, arthritis,
arthrosis, asthma, inflammatory bowel disease, Crohn's disease,
peritonitis, gastroesophageal reflux, uveitis, Guillain-Barre
syndrome, glomerulo-nephritis, lupus erythematosus and psoriasis.
The NOS-2 was also involved in the growth of certain forms of
tumors such as, for example, epitheliomas, adenocarcinomas or
sarcomas, and in infections with Gram-positive or Gram-negative
intracellular or extracellular bacteria.
[0012] In all the situations in which an overproduction of NO is
deleterious, it thus appears to be desirable to reduce the
production of NO by administering substances capable of inhibiting
the NOS-2. However, given the important physiological roles played
by the constitutive isoform NOS-3, in particular in regulating the
arterial pressure, it is essential that the inhibition of the
isoform NOS-2 has the least possible effect on the isoform NOS-3.
Actually, it is known that the administration of unselective
inhibitors of NOS isoforms leads to vasoconstriction and an
increase in arterial pressure (Moncada, S., Palmer, R. M. J. and
Higgs, E. A., Biosynthesis of nitric oxide from L-arginine: a
pathway for the regulation of cell function and communication,
Biochem. Pharmacol., 1989, 38: 1709-1715). These effects on the
cardiovascular system are deleterious since they reduce the supply
of nutrients to the tissues. Consequently, the present invention
relates to compounds whose inhibitory activity with respect to
NOS-2 is significantly higher than their inhibitory activity with
respect to NOS-3.
[0013] Thiazoline-based NOS inhibitors are described in particular
in patent applications WO94/12165, WO95/11231 and WO96/14842.
[0014] The present invention relates to the use of
2-amino-thiazoline derivatives of formula (I) in which: [0015]
either Y is a methylene (CH.sub.2) and X is chosen from the
following groups: O, NH, N--(C.sub.1-C.sub.4)alkyl, N-Bn, N-Ph,
N-(2-Py), N-(3-Py), N-(4-Py), N-2-pyrimidyl, N-5-pyrimidyl, S, SO,
SO.sub.2, CH.sub.2 or CHPh; [0016] or Y is a carbonyl (C.dbd.O) and
X is chosen from the following groups: NH, N-Ph, N-(2-Py),
N-(3-Py), N-(4-Py), N-2-pyrimidyl, N-5-pyrimidyl for the
preparation of medicinal products for preventing and treating
diseases in which an abnormal production of nitric oxide (NO) by
induction of inducible NO-synthase (NOS-2 or iNOS) is involved.
[0017] In the above definitions and in those which follow, the
alkyl radicals contain 1 to 4 carbon atoms in a straight or
branched chain. The abbreviations Bn, Py, Ph mean respectively
benzyl, pyridyl, phenyl.
[0018] The compounds of formula (I) contain one or more asymmetric
carbons and can thus be in racemic form or in the form of
enantiomers and diastereoisomers; these also form a part of the
invention as well as the mixtures thereof.
[0019] Moreover, the compounds of formula (I) can be in the
tautomeric form (Ia) ##STR3## These tautomers also form a part of
the invention.
[0020] Among the compounds of formula (I) useful according to the
invention, mention may be made of the following compounds: [0021]
4-(morpholin-4-ylmethyl)-4,5-dihydro-1,3-thiazol-2-ylamine, [0022]
4-(piperazin-1-ylmethyl)-4,5-dihydro-1,3-thiazol-2-ylamine, and
[0023]
4-(4-methyl-piperazin-1-ylmethyl)-4,5-dihydro-1,3-thiazol-2-ylamine,
[0024] the racemic mixtures, enantiomers, diastereoisomers,
tautomers thereof, as well as the pharmaceutically acceptable salts
thereof.
[0025] Among the compounds useful according to the invention and
particularly preferred, mention may be made of the following
compound: [0026]
4-(4-methyl-piperazin-1-ylmethyl)-4,5-dihydro-1,3-thiazol-2-ylami-
ne, [0027] the racemic mixtures, enantiomers, tautomers thereof, as
well as the pharmaceutically acceptable salts thereof.
[0028] The invention also relates to the pharmaceutical
compositions containing, as active plinciple, a derivative of
formula (I) for which either Y is a methylene (CH.sub.2) and X is
chosen from the following groups: O, NH, N--(C.sub.1-C.sub.4)alkyl,
N-Bn, N-Ph, N-(2-Py), N-(3-Py), N-(4-Py), N-2-pyrimidyl,
N-5-pyrimidyl, S, SO, SO.sub.2, CH.sub.2 or CHPh; or Y is a
carbonyl (C.dbd.O) and X is chosen from the following groups: NH,
N-Ph, N-(2-Py), N-(3-Py), N-(4-Py), N-2-pyrimidyl, N-5-pyrimidyl as
well as the racemic mixtures, enantiomers, diastereoisomers,
tautomer thereof, and pharmaceutically acceptable salts
thereof.
[0029] The compounds of formula (I) can be prepared by cyclization
of a derivative of formula (II): ##STR4## in which X and Y have the
same meaning as in formula (I).
[0030] This cyclization is generally carried out using an acid such
as hydrochloric acid, in aqueous medium, at a temperature of about
100.degree. C. 6N hydrochloric acid is generally used.
[0031] The derivatives of formula (II) can be obtained according to
the following reactions schemes: ##STR5## in these formulae, X and
Y have the same meanings as in formula (I), Ra is a protecting
group of the amine function such as those described by T. W.
GREENE, Protective groups in Organic Synthesis, J.
Wiley-Interscience Publication (1991), preferably an acetyl or
tert-butyloxycarbonyl radical, and Rb is a (C.sub.1-C.sub.4) alkyl
or alkoxycarbonyl radical, preferably methyl, ethyl or
isobutyloxycarbonyl.
[0032] The reaction a is generally carried out in the presence of a
Lewis acid such as the iron trichloride (III), in an inert solvent
such as dichloromethane or acetonitrile, at a temperature of
between 10.degree. C. and the boiling point of the reaction medium.
When X represents NH, X can be protected by a protecting group of
the amine function such as described by T. W. GREENE, Protective
Groups in Organic Synthesis, J. Wiley- Interscience Publication
(1991), preferably using a tert-butoxycarbonyl radical.
[0033] The reduction reaction b is preferably carried out using a
hydride such as sodium borohydride or lithium aluminum hydride in a
(C.sub.1-C.sub.4) aliphatic alcohol or tetrahydrofuran, at a
temperature of between 0.degree. C. and 30.degree. C.
[0034] The deprotection reaction c for the compounds in which Ra is
a protecting group of the amine function is carried out by any
deprotection method known to those skilled in the art and in
particular those described by T. W. GREENE, Protective Groups in
Organic Synthesis, J. Wiley-Interscience Publication (1991).
Preferably when the protecting group is an acetyl radical, this
reaction is carried out using aqueous hydrochloric acid at a
temperature of about 100.degree. C. When the protecting group is a
tert-butoxycarbonyl radical, this reaction is carried out using
hydrochloric acid in dioxane, at a temperature of about 20.degree.
C.
[0035] The reaction d is carried out by the action of tert-butyl
isothiocyanate, in an inert solvent such as (C.sub.1-C.sub.4)
aliphatic alcohol (preferably methanol or ethanol), optionally in
the presence of a tertiary amine such as triethylamine, at a
temperature between 20.degree. C. and the boiling point of the
reaction medium.
[0036] The compounds of formula (I) in which X represents either
SO, or SO.sub.2 can be obtained by direct oxidation of the compound
of formula (I) in which X represents S. This oxidation is carried
out according to the known methods of oxidation of organosufur
compounds, such as described by M. HUDLICKY, Oxidation in Organic
Chemistry, ACS Monograph, 186, 252-263 (1990). For example, it is
carried out by the action of an organic peracid or organic peracid
salt (percarboxylic or persulfonic acid, in particular perbenzoic
acid, 3-chloro-perbenzoic acid, 4-nitroperbenzoic acid, peracetic
acid, pertrifluoroacetic acid, performic acid, monopelphthalic
acid) or a mineral peracid (for example, periodic or persulfuric
acid), in an inert solvent such as a chlorine solvent (for example,
trichloroethane or dichloromethane), at a temperature of between
0.degree. C. and 20.degree. C. The hydrogen peroxide or periodate
(sodium periodate, for example), in an inert solvent such as
(C.sub.1-C.sub.4) aliphatic alcohol, water or a mixture of these
solvents, at a temperature between 0.degree. and 20.degree. C. can
also be used. These products can also be prepared from the
cortesponding compounds of formula (II), obtained according to the
following reaction schemes: ##STR6##
[0037] The oxidation reaction is carried out according to the known
methods of oxidation of organosulfur compounds as described
above.
[0038] The deprotection reaction b for the compounds in which Ra is
a protecting group of the amine function is carried out by any
method of deprotection known by those skilled in the art and
particularly those described by T. W. GREENE, Protective Groups in
Organic Synthesis, J. Wiley-Interscience Publication (1991).
Preferably when the protecting group is an acetyl radical, this
reaction is carried out using aqueous hydrochloric acid, at a
temperature of about 100.degree. C. When the protecting group is a
tert-butyloxycarbonyl radical, this reaction is carried out using
hydrochloric acid in dioxane, at a temperature of about 20.degree.
C.
[0039] The reaction c is carried out by the action of tert-butyl
isothiocyanate, in an inert solvent such as (C.sub.1-C.sub.4)
aliphatic alcohol (preferably methanol or ethanol), optionally in
the presence of a tertiary amine such as triethylamine, at a
temperature of between 20.degree. C. and the boiling point of the
reaction medium.
[0040] The compounds of formula (I) are isolated and can be
purified by the usual known methods, for example crystallization,
chromatography or extraction.
[0041] The enantiomers of the compounds of formula (I) can be
obtained by resolving the racemic mixtures, for example by
chromatography on a chiral column according to PIRCKLE W. H. et
al., Asymmetric Synthesis, Vol. 1, Academic Press (1983) or by
formation of salts or by synthesis from chiral precursors. The
diastereoisomers can be prepared according to the known
conventional methods (crystallization, chromatography or from
chiral precursors).
[0042] The compounds of formula (I) can optionally be converted to
addition salts with a mineral or organic acid by the action of such
an acid in an organic solvent such as an alcohol, a ketone, an
ether or a chlorinated solvent. These salts also form a part of the
invention.
[0043] Examples of pharmaceutically acceptable salts which may be
mentioned are the following salts: benzenesulfonate, hydrobromide,
hydrochloride, citrate, ethanesulfonate, fumarate, gluconate,
iodate, isethionate, maleate, methanesulfonate,
methylenebis-.beta.-oxynaphthoate, nitrate, oxalate, palmoate,
phosphate, salicylate, succinate, sulfate, tartrate,
theophyllinacetate and p-toluenesulfonate.
[0044] The compounds of formula (I) are inhibitors of NO-synthase
inducible or NO-synthase of type 2 (NOS-2) and are thus useful for
preventing and treating disorders associated with an excessive NO
production such as multiple sclerosis, focal or global cerebral
ischemia, cerebral or spinal trauma, Parkinson's disease,
Huntington's disease, Alzheimer's disease, amyotrophic lateral
sclerosis, migraine, depression, schizophrenia, anxiety, epilepsy,
diabetes, atherosclerosis, myocarditis, arthritis, arthrosis,
asthma, inflammatory bowel disease, Crohn's disease, peritonitis,
gastro-esophageal reflux, uveitis, Guillain-Barre syndrome,
glomerulo-nephritis, lupus erythematosus and psoriasis, the growth
of certain forms of tumors such as for example epitheliomas,
adenocarcinomas or sarcomas, and in infections with Gram-positive
or Gram-negative intracellular or extracellular bacteria.
[0045] Their activities as inhibitors of NOS-2 and NOS-3 were
determined by measuring the conversion of [.sup.3H]-L-arginine into
[.sup.3H]-L-citrulline with, respectively, a NOS-2 enzymatic
fraction extracted from the lungs of rats or mices pretreated with
lipopolysaccharides (10 mg/kg i.p. 6 hours before collecting the
tissue) and with a commercial preparation of recombinant bovine
NOS-3. The compounds were incubated for 20 to 30 minutes at
37.degree. C. in the presence of 5 .mu.M (for NOS-2 activity) or 10
.mu.M (for NOS-3 activity) of [.sup.3H]-L-arginine, 1 mM of NADPH,
15 .mu.M of tetrabiopterine, 1 .mu.M of FAD, 0.1 mM of DTT in a
HEPES buffer (50 mM, pH 6.7) containing 10 .mu.g/ml of calmodulin
and 1.25 mM of CaCl.sub.2 when the NOS-3 activity was measured. The
incubation was stopped by adding cold HEPES buffer (100 mM, pH 5.5)
containing 10 mM EGTA and 500 mg of cationic ion-exchange resin
(AG50W-X8, counter-ion: Na.sup.+) to separate the
[.sup.3H]-L-arginine from the [.sup.3H]-L-citrulline. After
separation of the phases by settling for 5 min, the radioactivity
remaining in the liquid phase was measured in a scintillation
counter in the presence of a suitable scintillation liquid. The
yield for the recovery of the formed L-[.sup.3H]citrulline was able
to be estimated using L-[ureido-.sup.14C]-citrulline as external
standard.
[0046] The NOS-2 or NOS-3 activity was expressed in picomole(s) of
[.sup.3H]-L-citrulline formed per minute and per milligram of
protein contained in the reaction medium.
[0047] In this test on the enzyme NOS-2, the IC.sub.50 value for
the compounds of formula (I) is less than or equal to 10 .mu.M.
[0048] The selectivity is measured by the IC.sub.50 NOS-3/IC.sub.50
NOS-2 ratio. This selectivity is greater than 45.
[0049] The compounds of formula (I) are of low toxicity. Their
LD.sub.50 is greater than 40 mg/kg via cutaneous route in mice.
[0050] The following examples illustrate the invention.
EXAMPLE 1
4-(4-Methyl-piperazin-1-ylmethyl)-4,5-dihydro-thiazol-2-ylamine
trihydrochloride
[0051] ##STR7##
[0052] A suspension of 0.42 g de
N-(tert-butyl)-N'-[2-hydroxy-1-(4-methyl-piperazin-1-ylmethyl)ethyl]-thio-
urea in 3.9 mL of an aqueous 6N hydrochloric acid is heated at a
temperature of about 100.degree. C. for 5 hours. After cooling, the
reaction medium is concentrated under reduced pressure (2 kPa) at a
temperature of about 55.degree. C. The residue obtained is dried in
an oven under vacuum (2 kPa) for 4 hours. About 0.47 g of
4-(4-methyl-piperazin-1-ylmethyl)-4,5-dihydro-thiazol-2-ylamine,
trihydrochloride are obtained in the form of a very hygroscopic
off-white paste. [.sup.1H NMR spectrum (300 MHz, (CD.sub.3).sub.2SO
d6 with addition of a few drops of CD.sub.3COOD d4. .delta.in ppm):
from 2.55 to 2.90 (mf, 4H); 2.80 (s, 3H); from 2.95 to 3.30 (mf,
4H): from 3.30 to 3.60 (mf, 2H); 3.40 (dd, J=11.5 and 5.5 Hz, 1H);
3.69 (dd, J=11.5 and 7.5 Hz, 1H); 4.51 (mt, 1H)].
N-(tert-Butyl)-N'-[2-hydroxy-1-(4-methyl-piperazin-1-ylmethyl)ethyl]-thiou-
rea
[0053] ##STR8##
[0054] To a solution of 1 g of
2-amino-3-(4-methyl-piperazinl-yl)-1-propanol hydrochloride in 20
mL of absolute ethanol and 1.43 mL of triethylamine, about 0.78 mL
of tert-butylisothiocyanate are added. The reaction mixture is
stirred under inert atmosphere at a temperature of about 20.degree.
C. for 42 hours then is heated at a temperature of about 50.degree.
C. for 1 hour 30 min. After cooling at a temperature of about
20.degree. C., the reaction medium is evaporated under reduced
pressure (2 kPa) at a temperature of about 30.degree. C. The
residue thus obtained is taken up in 10 mL of water and 40 mL of
dichloromethane. The aqueous phase is extracted with 2 times 30 mL
of dichloromethane. The organic phases are collected, washed with
15 mL of water, dried over magnesium sulfate, filtered, then
concentrated under reduced pressure (2 kPa) at a temperature of
about 20.degree. C. About 0.42 g of
N-(tert-butyl)-N'-[2-hydroxy-1-(4-methyl-piperazin-1-ylmethyl)ethyl]thiou-
rea are obtained in the form of a white paste. [Infrared spectrum
between lamella of KBr 3279; 3075; 2939; 2806; 1533; 1459; 1359;
1295; 1204; 1010 and 821 cm.sup.-1].
2-Amino-3-(4-methyl-piperazin-1-yl)-1-propanol hydrochloride
[0055] ##STR9##
[0056] A suspension of 0.89 g of
N-[2-hydroxy-1-(4-methyl-piperazin-1- ylmethyl)ethyl] acetamide in
10.3 mL of an aqueous acid solution of 6N hydrochloric acid is
heated at a temperature of about 100.degree. C. for 3 hours. After
cooling at a temperature of about 60.degree. C., the reaction
medium is filtered and the filtrate is concentrated under reduced
pressure (2 kPa) at a temperature of about 60.degree. C. About 1 g
of 2-amino-3-(4-methyl-piperazinyl)-1-propanol, hydrochloride is
obtained in the form of a tacky beige-colored paste. [Infrared
spectrum (KBr) 3337; 2955; 2637; 2522; 1617; 1457; 1062; 1009 and
962 cm.sup.-1].
N-[2-Hydroxy-1-(4-methyl-piperazin-1-ylmethyl)ethyl]acetamide
[0057] ##STR10##
[0058] A solution under inert atmosphere of 3.27 g of methyl
(acetylamino)-3-(4-methyl-piperazin-1-yl)propanoate in 100 mL of
anhydrous methanol is cooled at a temperature of about 10.degree.
C., then 0.76 g of sodium borohydride are added using a spatula.
The reaction medium is stirred for 5 hours at a temperature of
about 20.degree. C., then are added again 0.26 g of sodium
borohydride and the stirring is carried out for 38 hours. Then, 5
mL of water is dropped into the reaction mass which is heated and
concentrated under reduced pressure (2 kPa) at a temperature of
about 30.degree. C. The obtained residue is taken up with
dichloromethane and the insoluble matter is removed by filtration.
The filtrate is concentrated under reduced pressure (2 kPa) at a
temperature of about 20.degree. C. The residue is purified by
chromatography under argon pressure (60 kPa), on a column of silica
gel (particle size 40-63 .mu.m; diameter 5 cm; height 19 cm),
eluting with successive mixtures of 20%
dichloromethane/methanol/aqueous ammonia (98/2/0, 95/5/0.1,
90/10/0.2, 80/20/0.25, 50/50/0.25 by volume). The fractions
containing the expected product are combined and concentrated under
reduced pressure (2 kPa) at a temperature of about 40.degree. C.
About 0.92 g of
N-[2-hydroxy-1-(4-methyl-piperazin-1-ylmethyl)ethyl] acetamide are
obtained in the form of a yellow-colored liquid. [Infrared spectrum
CH.sub.2Cl.sub.2 3621; 3429; 3352; 2944; 2803; 1657; 1513; 1460;
1284; 1050; 1011 and 816 cm.sup.-1].
Methyl 2-(Acetylamino)-3-(4-methyl-piperazin-1-yl)propanoate
[0059] ##STR11##
[0060] To a solution of 8.57 g of methyl 2-acetamidoacrylate in 500
mL of dichloromethane stirred under inert atmosphere, about 6.65 mL
of N-methylpiperazine are added, then 0.97 g of iron trichloride
are added, and the mixture is stirred at a temperature of about
20.degree. C. for 66 hours. Then, 300 mL of an aqueous solution of
sodium sulfate are dropped to the reaction medium while stirring
the reaction mixture and the mixture filtered through Celite. After
separation of the phase by settling, the organic phase is dried
over sodium sulfate, filtered and then concentrated in a vacuum
oven under reduced pressure (2 kPa) at a temperature of about
40.degree. C. in order to obtain an orange-colored liquid. The
aqueous phase is extracted with 3 times 150 mL of dichloromethane
and all of the organic extracts are collected, dried over sodium
sulfate, then concentrated under reduced pressure (2 kPa) at a
temperature of about 20.degree. C. in order to obtain an yellow
oil. Both of the organic extracts as described above are combined
and purified by chromatography under argon pressure (50 kPa), on a
column of silica gel (particle size 40-63 .mu.m; diameter 5 cm;
height 25 cm), eluting with successive mixtures of 20%
dichloromethane/methanol/aqueous ammonia (99/1/0, 97/3/0,
90/10/0.25, 80/20/0.25 by volume). The fractions containing the
expected product are combined and concentrated under reduced
pressure (2 kPa) at a temperature of about 30.degree. C. About 3.3
g of methyl 2-(acetylamino)-3-(4-methyl-piperazinyl)propanoate are
obtained in the form of a yellow liquid. [Infrared spectrum
CCl.sub.4 3437; 3318; 2941; 2798; 1749; 1685; 1499; 1458; 1374;
1286; 1204; 1168 and 1014 cm.sup.-1]
[0061] The pharmaceutical compositions according to the invention
consist of a compound of formula (I) or an isomer or tautomer or
salt of such a compound, in pure form or in the form of a
composition in which it is combined with any other pharmaceutically
compatible product, which may be inert or physiologically active.
The medicinal products according to the invention may be used
orally, parenterally, rectally or topically.
[0062] Solid compositions for oral administration which can be used
include tablets, pills, powders (gelatin capsules, cachets) or
granules. In these compositions, the active principle according to
the invention is mixed with one or more inert diluents, such as
starch, cellulose, sucrose, lactose or silica, under a stream of
argon. These compositions can also comprise substances other than
diluents, for example, one or more lubricants such as magnesium
stearate or talc, a dye, a coating (dragees) or a varnish.
[0063] Liquid compositions for oral administration which can be
used include pharmaceutically acceptable solutions, suspensions,
emulsions, syrups and elixirs containing inert diluents such as
water, ethanol, glycerol, plant oils or liquid paraffin. These
compositions can comprise substances other than diluents, for
example, wetting products, sweeteners, thickeners, flavorings or
stabilizers.
[0064] The sterile compositions for parenteral administration can
preferably be aqueous or non-aqueous solutions, suspensions or
emulsions. Solvent or vehicles which may be used include water,
propylene glycol, a polyethylene glycol, plant oils, in particular,
olive oil, injectable organic esters, for example ethyl oleate, or
other suitable organic solvents. These compositions can also
contain adjuvants, in particular, wetting agents, solvents. These
compositions can also contain adjuvants, in particular, wetting
agents, isotonic agents, emulsifiers, dispersants and stabilizers.
The sterilization can be carried out in several ways, for example,
by aseptic filtration, by incorporating sterilizing agents into the
composition, by irradiation or by heating. They can also be
prepared in the form of sterile solid compositions which can be
dissolved at the time of use in sterile water or any other
injectable sterile medium The compositions for rectal
administration are suppositories or rectal capsules which contain,
besides the active product, excipients such as cocoa butter, semi-
synthetic glycerides or polyethylene glycols.
[0065] The compositions for topical administration can be, for
example, creams, lotions, eye drops, mouth washes, nasal drops or
aerosols.
[0066] In human therapy, the compounds according to the invention
are particularly useful for treating and/or preventing multiple
sclerosis, focal or global cerebral ischemia, cerebral or spinal
trauma, Parkinson's disease, Huntington's disease, Alzheimer's
disease, amyotrophic lateral sclerosis, migraine, depression,
schizophrenia, anxiety, epilepsy, diabetes, atherosclerosis,
myocarditis, arthritis, arthrosis, asthma, inflammatory bowel
disease, Crohn's disease, peritonitis, gastro-esophageal reflux,
uveitis, Guillain-Barre syndrome, glomerulo-nephritis, lupus
erythematosus, psoriasis, the growth of certain forms of tumors
such as, for example, epitheliomas, adenocarcinomas or sarcomas,
and in infections with Gram-positive or Gram-negative intracellular
or extracellular bacteria.
[0067] The doses depend on the desired effect, the duration of the
treatment and the route of administration used; they are generally
comprised between 1 mg and 100 mg per day via the oral route for an
adult, with unit doses ranging from 0.5 mg to 50 mg of active
substance.
[0068] The examples which follow illustrate compositions according
to the invention:
EXAMPLE A
[0069] Gel capsules containing 50 mg of active product and having
the composition below are prepared, according to the usual
technique: TABLE-US-00001 Compound of formula (I) 50 mg Cellulose
18 mg Lactose 55 mg Colloidal silica 1 mg Sodium
carboxymethylstarch 10 mg Talc 10 mg Magnesium stearate 1 mg
EXAMPLE B
[0070] Tablets containing 50 mg of active product and having the
composition below are prepared, according to the usual technique:
TABLE-US-00002 Compound of formula (I) 50 mg Lactose 104 mg
Cellulose 40 mg Polyvidone 10 mg Sodium carboxymethylstarch 22 mg
Talc 10 mg Magnesium stearate 2 mg Colloidal silica 2 mg Mixture of
hydroxymethylcellulose, glycerol, titanium oxide (72/3.5/24.5) q.s.
1 finished film-coated tablet weighing 245 mg.
EXAMPLE C
[0071] An injectable solution containing 10 mg of active product
having the following composition: TABLE-US-00003 Compound of
formula (I) 10 mg Benzoic acid 80 mg Benzyl alcohol 0.06 ml Sodium
benzoate 80 mg 95% ethanol 0.4 ml Sodium hydroxide 24 mg Propylene
glycol 1.6 ml Water q.s 4 ml
[0072] The present invention also relates to the method for
preventing and treating diseases in which an abnormal production of
nitric oxide (NO) by induction of inducible NO-synthase (NOS-2 or
iNOS) is involved by administration of a compound of formula (I),
the racemic mixtures, enantiomers, diastereoisomers thereof and
mixtures thereof, tautomer thereof and pharmaceutically acceptable
salts thereof.
* * * * *