U.S. patent application number 09/367579 was filed with the patent office on 2001-07-19 for (-)e-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethyl-piperidine, and its antidiarrhoeal use.
Invention is credited to CALVET, ALAIN, CHOVET, MARIA, DAHL, SVEIN, JACOBELLI, HENRY, RIVIERE, PIERRE, THEODOROU, VASSILIA.
Application Number | 20010008897 09/367579 |
Document ID | / |
Family ID | 9504511 |
Filed Date | 2001-07-19 |
United States Patent
Application |
20010008897 |
Kind Code |
A1 |
CALVET, ALAIN ; et
al. |
July 19, 2001 |
(-)E-2-(3,4-DICHLOROCINNAMYL)-1-CYCLOPROPYLMETHYL-PIPERIDINE, AND
ITS ANTIDIARRHOEAL USE
Abstract
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethyl-piperidine of
formula (I) and its addition salts with pharmaceutically acceptable
acids; their use as antidiarrhoeals in mammals; pharmaceutical
compositions intended for treatment of diarrhoeas.
Inventors: |
CALVET, ALAIN; (ANN ARBOR,
MI) ; CHOVET, MARIA; (MONTROUGE, FR) ; DAHL,
SVEIN; (TROMSDALEN, NO) ; JACOBELLI, HENRY;
(PARAY-VIEILLE-POSTE, FR) ; THEODOROU, VASSILIA;
(PORTET-SUR-GARONNE, FR) ; RIVIERE, PIERRE; (SAN
DIEGO, CA) |
Correspondence
Address: |
CHARLES W ASHBROOK
WARNER LAMBERT COMPANY
2800 PLYMOUTH ROAD
ANN ARBOR
MI
48105
|
Family ID: |
9504511 |
Appl. No.: |
09/367579 |
Filed: |
November 10, 1999 |
PCT Filed: |
March 4, 1998 |
PCT NO: |
PCT/EP98/01564 |
Current U.S.
Class: |
514/317 ;
546/192 |
Current CPC
Class: |
C07D 211/18 20130101;
A61P 1/12 20180101 |
Class at
Publication: |
514/317 ;
546/192 |
International
Class: |
A61K 031/445 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 5, 1997 |
FR |
97/02713 |
Claims
1. (-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperidine of
formula (I) 3and its addition salts with pharmaceutically
acceptable acids.
2. The hydrochloride of
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylp- iperidine
(I).
3. Process for the preparation of
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopro- pylmethylpiperidine (I)
which consists: in carrying out the resolution of (+/-)
E-2-(3,4-dichlorocinnamyl)piperidine via the eutomeric
diastereoisomeric salt with N-acetyl-L-phenylalanine which leads to
(+) E-2-(3,4-dichlorocinnamyl)piperidine (III) and then, in
acylating the compound (III) with cyclopropanecarbonyl chloride to
obtain (+)
E-2-(3,4-dichlorocinnamyl)-1-cyclopropanecarbonylpiperidine (II)
and then, in reducing the compound (II) with a metallic or
organometallic hydride to obtain
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperi- dine (I)
and optionally in making the addition salt with a pharmaceutically
acceptable acid.
4. As medicament, (-)
E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperi- dine (I) and
its addition salts with pharmaceutically acceptable acids.
5. Therapeutic composition containing by way of active principle
(-) E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperidine (I)
and its addition salts with pharmaceutically acceptable acids.
6. Use of
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperidine (I) or
of one of its pharmaceutically acceptable salts for the preparation
of antidiarrhoeal medicaments.
7. As novel industrial products, (+)
E-2-(3,4-dichlorocinnamyl)piperidine (III) and (+)
E-2-(3,4-dichlorocinnamyl)-1-cyclopropanecarbonylpiperidine (II) as
prepared in claim 3.
Description
FIELD OF THE INVENTION
[0001] The invention relates to
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropy- lmethylpiperidine, its
salts, and their use as antidiarrhoeal medicaments in man.
PRIOR ART
[0002] The secretory anomalies of the gastro-intestinal tract are
responsible, with motor disorders, for the majority of chronic or
acute diarrhoeas which, in 1990, were estimated to be the second
cause of global mortality, especially in child populations of
developing countries.
[0003] Chronic diarrhoeas are defined by their generally persistent
duration of over two weeks. Their known mechanisms and the
diagnostic strategy to be adopted in view of these cases has been
documented by M. CERF Gastroenterol. Clin. Biol, 1992, 16, T 12-T
21. and more recently by M. J. G. FARTHING--Eur. J. of
Gastroenterol. & Hepatol. 1996, 8:157-167. The acute
diarrhoeas, a great majority of which are of infectious origin,
have likewise been documented by M. CERF and M. HAGIAGE: Diarrhes
aigus d'origine infectieuse (Acute diarrhoeas of infectious
origin),--Editions Techniques--Encycl. Md. Chir. (Paris-France),
Gastro-entrologie, 9061 A.sup.10, 1992, 20pp.; and H. L.
DuPONT--Review article: infectious diarrhoea--Aliment. Pharmacol.
Ther. 1994; 8: pp.3-13. Among other causes, the important role of
toxinogenesis during bacterial infection is discussed, and,
especially, the expression of the pathogenic capacity by the
synthesis of thermolabile or thermostable cytotoxins and
enterotoxins, which are responsible for secretory diarrhoeas with a
hydroelectrolytic component, and whose representative
physiopathological model is that of cholera. Other infectious
agents are known to cause diarrhoeas of this type, such as
Salmonella, Escherichia Coli (E. coli) and Clostridium difficile
(C. difficile) strains.
[0004] These latter agents, and more particularly C. difficile, are
responsible for chronic and abundant secretory diarrhoeas, often of
nosocomial origin, in subjects submitted to an intensive antibiotic
therapy such as HIV positive patients. In the latter, the
particularly incapacitating diarrhoeas are often associated with
malabsorption, and contribute to the rapid development of an
alarming state of denutrition.
[0005] For the treatment of secretory diarrhoeas, rehydration of
the patients is recommended and sometimes turns out to be
essential. Some compounds have been shown to be active
(phenothiazine, chlonidine, bismuth salts) but their sensitive
employment because of their secondary effects has led to their
generalization being abandoned. The usual symptomatic treatments
call for adsorbent compounds (Fuller's earth), modulators of the
intestinal flora and, very widely, compounds called retardants,
which are morphinomimetic antidiarrhoeals: loperamide (INN) and
diphenoxylate (INN), known inhibitors of the motility of the GI
tract: and, in fact, of controversial if not inadvisable utility
for certain ailments, among other reasons through the delay which
they contribute to the natural evacuation of pathogenic
bacteria.
[0006] More recently, it has been proposed to treat these
diarrhoeas with acetorphan (INN), a synthetic enkephalinase
inhibitor dipeptide with antisecretory effect, which maintains the
effect of enkephalins, antisecretory endogenous neuropeptides of
the intestinal wall, which are normally rapidly hydrolysed in vivo
by the enkephalinases which makes their effect fleeting.
[0007] As far as the therapy of diarrhoeas of patients infected
with HIV is concerned, it is frequently necessary to resort to
serious methods, which can only be carried out in an inpatient
environment, such as rehydration and renutrition by the enteral or
parenteral routes, which are combined with the symptomatic
antidiarrhoeal treatment and an antibiotic therapy directed against
the possible pathogenic agent. The usual antidiarrhoeal agents only
have, most often, a relative and episodic efficacy. Recently, for
these ciiarrhoeas and, more generally, cases resistant to
conventional therapy, peptides inhibiting motility and
gastrointestinal secretion related to somatostatin have been
proposed (M. CAMILLERI--Digestion 1996;57 (suppl 1): 90-92 and M.
J. G. FARTHING--Digestion 1996;57 (suppl 1): 107-113). Substitute
synthetic compounds for this endogenous mediator are octreotide
(INN) and valtreotide (INN), both octapeptides proposed with some
success for the treatment of secretory diarrhoeas of AIDS. Although
their duration of action is considerably longer than that of
somatostatin, these expensive compounds are only active by repeated
administration parenterally which leads to prohibitive treatment
costs and, because of their mode of administration, makes their use
virtually impossible in an outpatient environment. In addition,
their lack of specificity, which has been pointed out, can involve
secondary effects which dramatically aggravate the state of
denutrition of the patients (disorders of the regulation of
hydrocarbon metabolism and increase in steatorrhoea).
[0008] In addition, certain compounds defined as specific ligands
for sigma receptors have shown antisecretory properties suggesting
their use in the treatment of diarrhoeas. Thus
(+)-N-cyclopropylmethyl-N-methyldiph-
enyl-1,4-ethyl-1-buten-3-yl-1-amine, or igmesine (INN), and its
hydrochloride are disclosed among other compounds in European
Patent 0 362 001. The compounds of this patent are defined in vitro
as specific ligands for sigma receptors and shown, in vivo, in
rats, to be inhibitors of amnesic phenomena caused by scopolamine,
and inhibitors of gastroduodenal ulcers caused by the
administration of cysteamine, this last activity being connected
with their capacity to increase the alkaline duodenal secretion in
anaesth,etized animals. In the broad sense, the compounds of this
patent are indicated as useful for the treatment of dysfunctions of
the gastrointestinal tract such as disorders of peristalsis, of
motility, the phenomena of gastro-oesophageal and gastroduodenal
reflux as well as for gastric and gastroduodenal ulceration.
[0009] Subsequently to these studies, the sigma receptors, whose
localization was known in the central nervous system and the immune
system, have been demonstrated by F. ROMAN et al. in the
gastrointestinal tract of the guinea-pig (Life Sciences 1988, 42,
2217-2222) and then of man (Gastroenterology 1991, 100, A662).
[0010] In connection with these localizations, various experiments,
among others by J. L. JUNIEN et al. (Neuropharmacology, Volume 30,
No. 10, October 1991, pp. 1119-1124) have demonstrated the
inhibitory action of igmesine on colonic hypermotility induced by
stress via the corticotropin releasing hormone (CRH or CRF) in man.
In addition,, P. RIVIERE et al. (Gastroenterol. Clin. Biol. 1991,
15 (2B), A70) show in vitro that igmesine modifies the
transmembrane ionic transport through portions of mouse jejunum.
This effect, antagonized by haloperidol, involves the participation
of sigma receptors. V. J. CARLISI et al. (FASEB J. 1992, 6 (4),
A1287) studied the effect of igmesine in vivo in mice, in a model
of inflammatory diarrhoea caused by PGE.sub.2: at a dose of 30
mg/kg, igmesine, co-administered by the i.p. route with PGE.sub.2,
delayed by approximately 15 min the appearance of the diarrhoea, an
inhibitory effect antagonized by haloperidol, and which, tested for
by the oral route, turned out to be zero at a dose of 60 mg/kg.
More recently, G. SHI et al. (UEGW 1996--Paris--abstract No. 0786)
showed the effect of igmesine at a dose of 200 mg p.o. in man on
intestinal hypersecretion induced by PGE.sub.2.
[0011] On the other hand, the application WO 95/15948 discloses
derivatives of 2-arylalkenylazacycloalkanes as ligands for sigma
receptors, a process for their preparation and their application in
therapeutics. The compounds, their isomers and their addition salts
are proposed for the preparation of antipsychotic medicaments and
are useful in gastroenterology. The experimental section describes
in Example 2E racemic
E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperidine and its
hydrochloride and, in addition, mentions without specific
experimental results that the compounds of the application are
active on secretory diarrhoeas induced in mice by Salmonella
lipopolysaccharicle (LPS), which suggests their use in the
treatment of secretory diarrhoeas of varying aetiologies.
[0012] Overcoming the difficulties and uncertainties of the prior
art; as set out, the present invention proposes for the purposes of
symptomatologic treatment of secretory diarrhoeas the use, in
appropriate medicamentous forms, of a novel optically active
compound, which is a ligand for sigma receptors, and whose spectrum
of antidiarrhoeal properties is particularly remarkable and
distinguishes it formally from the prior art.
SUMMARY OF THE INVENTION
[0013] The subject of the invention by way of novel compound is
optically pure
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperidine (I) of
formula 1
[0014] its addition salts with pharmaceutically acceptable acids
and a process for their preparation.
[0015] It likewise relates, by way of medicaments, to
(-)E2-(3,4-dichlorocinnamyl)-l-cyclopropylmethyl-piperidine (I) and
its, addition salts as well as their use in the preparation of
pharmaceutical compositions intended for the treatment of
diarrhoeas. It also comprises the medicamentous compositions
comprising by way of active principle
(-)E-2(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperidine (I) or
its addition salts in therapeutically efficacious quantity.
DETAILED DESCRIPTION OF THE INVENTION
[0016] In first place, the invention is directed at optically pure
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperidine (I) and
its addition salts with pharmaceutically acceptable acids.
[0017] Optically pure is understood as meaning that the product is
practically free of its optical antipode and is at least of an
optical purity of 95% and, preferably, equal to or greater than 98%
in levorotatory eutomer, this being determined by appropriate
analytical means.
[0018] Pharmaceutically acceptable addition salts are understood as
meaning those inorganic or organic salts, and their possible
isomers, shown to be non-toxic in the therapeutically customary
doses of which, for example, a list is presented in J. Pharm. Sci.,
1977, Volume 66, pp. 119. Non-limiting examples are acetic,
benzenesulphonic, camphorsulphonic, citric, ethanesulphonic,
hydrdbromic, lactic, maleic, malic, methanesulphonic, mucic,
nitric, pamoic, phosphoric, salicylic, stearic, succinic, sulphuric
or tartaric acid and hydrochloric acid, which is preferred. In
another aspect, the invention relates to a process for the
preparation of optically pure (-)E-2-(3,4-dichlorocinnamyl)-1-cyc-
lopropylmethylpiperidine (I) which consists either in resolving the
corresponding racemic compound described in Example 2E of the
application WO 95/15948, or in carrying out the chemical synthesis
starting from (+) E-2-(3,4-dichlorocinnamyl)piperidine (III),
itself obtained, according to the application WO 95/15948, by
resolution of the corresponding racemic compound described in
preparation 2E of the same patent application.
[0019] Resolution of the racemic compound corresponding to the
product of the invention consists in using an optically active acid
to obtain, with the racemate, diastereoisomeric addition salts
which are separated by crystallization, and from which the two
resolved enantiomers are generated by appropriate treatment. Acids
currently used for the preparation of such salts are, as
non-limiting examples, the enanticimers of .alpha.-phenylglycine,
.alpha.-phenylalanine, malic, mandelic and tartaric acids, of
camphanic acid or alternatively of
.alpha.-methoxy-.alpha.-trifluoromethylacetic acid. An alternative
method of resolution is the direct resolution of the racemic
compound by high-performance liquid chromatography on a column
containing, for example, as stationary phase a cellulose polymer
grafted with carbamate groups such as the phase CHIRACEL OD
(Daicel) and carrying out an elution with hexane containing a small
quantity of triethylamine.
[0020] However, the preferred process is an adaptation of the
methodology described in the application WO 95/15948 and consists
in carrying out the resolution of (+/-)
E-2-(3,4-dichlorocinnamyl)piperidine via the eutomeric
diastereoisomeric salt with N-acetyl-L-phenylalanine, which,
purified and treated, leads to (+) E-2-(3,4-dichlorocinnamyl)
piperidine (III), and then in acylating (III) with
cyclopropanecarboxylic acid to obtain (+)
E-2-(3,4-dichlorocinnamyl)-1-cyclopropanecarbonylpiperidine (II),
and then, in reducing (II) with a metallic or organometallic
hydride to obtain
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclo-propylmethylpiper- idine (I)
of suitable optical purity and, optionally, in making a
pharmaceutically acceptable salt: 2
[0021] At the resolution stage, the recovery and the purification
of the (-) antipode of the intermediate compound (III) allows,
according to the same process, the distomer of the compound of the
invention to be prepared, namely
(+)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperid- ine
synthesized as comparison product.
[0022] The compound which is the subject of the invention and its
salts have remarkable pharmacological properties, indicative of
their usefulness in the form of medicaments for the treatment of
secretory diarrhoeas in man. Although in vitro its affinity for
sigma receptors is not stereospecific but of comparable intensity
to that of its antipode, totally unexpectedly, the
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmeth- ylpiperidine
eutomer (I) shows in vivo, in various representative models of
toxigenic secretory diarrhoea, a very strong and stereoselective
antidiarrhoeal activity.
[0023] Thus, the levorotatory eutomer appears from 4 to more than
50 times more active, by the oral route in mice, than its parent
racemate in the models of secretory diarrhoea caused by Salmonella
lipopolysaccharide (LPS), the thermostable toxin of E. coli and the
A and B toxins of C. difficile, without acting on the transit.
Likewise, the eutomer turns out to be close to 50 times more active
than the racemate by the oral route, in rats, on the inhibition of
the intestinal secretion caused by cholera toxin.
[0024] In mice, by the oral route in the models of toxigenic
secretory diarrhoeas which have just been presented, when it is
compared to published compounds or those potentially capable of
treating secretory diarrhoeas, the eutomer which is the subject of
the invention turns out to be:
[0025] from 66 to more than 4000 times more active than
loperamide,
[0026] from 75 to more than 1400 times more active than
igmesine,
[0027] and, on diarrhoea caused by LPS, more than 6000 times more
active than acetorphan.
[0028] Finally, compared to the enantiomers described in the
application WO 95/15948 and which turn out to be equally active in
vivo in diarrhoea caused by LPS, the study as a function of the
dose shows, for the comparison products (-)
E-2-cinnamyl-1-cyclopropylmethylpyrrolidine (Example 1.3 of WO
95/15948) and (-) E-2-cinnamyl-l-cyclopropylmethyl-pip- eridine
(Example 2A.3), a progressive effect which, after a maximum,
decreases significantly, whereas, differently, the eutomer shows a
progressive effect followed by a stage in which this activity is
maintained, which is demonstrative of a safety of medical use and
of a therapeutic aid, contrary to the comparison products for which
the zone of active concentrations, or therapeutic window, is narrow
and leads to a dosage which is difficult to handle and thus to a
risk of inefficacy.
[0029] These studies, expanded on in the experimental section,
demonstrate the particularly interesting antidiarrhoeal activity of
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropyl-methylpiperidine (I)
and of its salts and their utility in the preparation of
pharmaceutical forms which can be administered to mammals including
humans by routes appropriate to the pathology and condition of the
subject. Thus, the medicaments can be administered by the
parenteral, transdermal or transmucosal route in conventionally
known forms. However, the pharmaceutical forms adapted to
outpatient treatments are preferred and especially those intended
for administration by the oral route.
[0030] The pharmaceutical compositions according to the invention
which contain a therapeutically efficacious quantity of compound
(I) or of one of its salts, are appropriate for the treatment of
diarrhoeas which may be commonplace, such as those of infants or of
travellers, and which may be acute and/or persistent and of varying
aetiology in which the secretory component can just as well result
from a decrease in absorption as from intestinal hypersecret
ion.
[0031] Thus, the compositions of the compound (I) are indicated for
the treatment of diarrhoeas of inflammatory origin (Crohn's
disease, post-radiotherapy enterites), of obstructions by lymphoid
hyperplasia or alternatively of anti-cancer chemotherapy.
[0032] Likewise, these compositions are appropriate for the
symptomatic treatment of hypersecretory diarrhoeas such as those
following neuroendocrine tumour conditions (Zollinger-Ellison
syndrome, VIPoma, somatostatinoma, carcinoid syndrome), of viral,
including HIV, or bacterial infections, or even of congenital
dysfunctions or those caused by cathartic drugs, and during
hypersecretions of intestinal inflammatory syndromes.
[0033] The good tolerance to the product shown in the preliminary
tests justifies, for treatments of two to three weeks, a daily
dosage of 5 to 50 mg and, in exceptional cases, for aggressive
treatments of short duration up to 100 mg. However, the majority of
diarrhoeal conditions treated are improved by daily dosages of 10
to 30 mg, the product being administered by the oral route, divided
into two to four administrations per 24 hours.
[0034] The product is administered in various pharmaceutical forms,
containing per unit from 1.25 to 25 mg of the compound (I) or of
one of its salts, especially of its hydrochloride; these forms can
be, as non-limiting examples, tablets, coated tablets, capsules,
gelules, powders, solutions, suspensions or gels.
[0035] For the so-called solid forms, the compound (I) or its salt
may represent from 1 to 90% by weight of the finished form, the
pharmaceutically acceptable excipients representing from 99 to 10%.
For liquid forms or those considered as such, the active principle
can represent from 0.1 to 10% by weight of the finished form, the
liquid phase representing from 99.9 to 90% by weight.
EXPERIMENTAL SECTION
EXAMPLE 1
[0036] (-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperidine
(I) and its hydrochloride Stage .alpha.): (+)
E-2-(3,4-dichlorocinnamyl)piperidin- e (III)
[0037] The compound is prepared by resolution of (+/-)
E-2-(3,4-dichlorocinnamyl)piperidine via the eutomeric
diastereoisomeric salt obtained with N-acetyl-L-phenylalanine. In a
500 ml round-bottomed flask equipped with a stirrer and a condenser
in the reflux position, 23.00 g (85.1 mmol) of the racemic compound
(preparation E2 of WO 95/15948) and 8.82 g (42.6 mmol) of
N-acetyl-L-phenylalanine are introduced into 325 ml of acetone.
[0038] The mixture is heated to reflux with stirring, and a
solution is obtained which is progressively cooled and then kept at
20.degree.C. for 16 hours with stirring. The eutomeric salt is
filtered (dry weight: 15.50 g-m.p.=134-141.degree.C.), the filtrate
is rendered alkaline and extracted with dichloromethane, and the
residual oil from concentration (13.60 g - 50.2 mmol) is salified
in the same manner as above in 200 ml of acetone with 6.25 g (30.1
mmol) of Nacetyl-D-phenylalanine. The distomeric salt is removed
and the filtrate is treated by rendering alkaline to obtain 7.15 g
(26.5 mmol) of residual product which is salified in 150 ml of
acetone with 3.57 g (17.2 mmol) of N-acetyl-L-phenylalanine. A
second supply of eutomeric salt 6.00 g m.p. 150-152.degree.C. is
thus obtained after filtration.
[0039] The combined eutomeric salts (21.50 g) are recrystallized
for purification in 60 ml of boiling water; 4.30 g of product
recrystallize at 20.degree.C. (m.p.=151-152.degree.C.)
[0040] The insoluble matter is recrystallized in 60 ml of boiling
water; 10.9 g of product (m.p.=153-154.degree.C.) recrystallize at
20.degree.C. The salt is suspended in water, rendered alkaline with
a solution of sodium hydroxide and extracted with dichloromethane.
The solvent is removed by distillation in vacuo and on a water
bath. 6.10 g of product are obtained in the form of a pale yellow
oil. Yield=53% [.alpha.].sub.D.sup.20=+9.600 (c=0.5;
dichloromethane)
[0041] TLC and NMR in accordance (identical to the racemic starting
product)
[0042] Stage b):
(+)E-2-(3,4-dichlorocinnamyl)-1-cyclopropane-carbonylpipe- ridine
(II)
[0043] In a 250 ml reactor equipped with a stirrer and protected
from moisture, 6.00 g (22.2 mmol) of (+)
E-2-(3,4-dichlorocinnamyl)piperidine (III) obtained in the previous
stage are introduced under a nitrogen atmosphere into 100 ml of
dichloromethane dried over molecular sieves. 3.40 ml or 2.47 g
(24.4 mmol) of triethylamine are added to the pale yellow solution
and, after cooling to 10.degree.C., 2.20 ml or 2.55 g (24.4 mmol)
of cyclopropanecarbonyl chloride are added in 10 min. The mixture
is kept with stirring for 30 minutes at 20-25.degree.C. and then
extracted successively with 100 ml of 10% ammonia, 100 ml of 10%
hydrochloric acid, 100 ml of saturated sodium bicarbonate solution
and finally 100 ml of water. After drying over sodium sulphate, the
dichloroimethane is removed by distillation in vacuo and on a water
bath. The residual product is obtained in the pure state as a
yellow oil. Weight: 7.50
g-yield=100%-[.alpha.].sub.D.sup.20=+22.600 (c=0.5;
dichloromethane)
[0044] TLC(CH.sub.2Cl.sub.2/MeOH 10% ammoniacal 90/10 v/v):
Rf=0.90-1.00
[0045] Stage c):
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropyl-methylpiperid- ine
(I)
[0046] Under a nitrogen atmosphere, protected from moisture and
without exceeding 0.degree.C., on the one hand a suspension of 2.65
g (69.5 mmol) of lithium aluminum hydride (LAH) is prepared in 40
ml of dry THF, and on the other hand a solution of 2.99 g (22.4 m
mol) of aluminum chloride in 40 ml of dry diethyl ether. After 30
min contact for each preparation, the LAH/THF suspension is
introduced in 10 min ago at 0.degree.C. into the ethereal solution
of AlCl.sub.3, then a solution of 7.40 g (21.9 mmol) of the amide
(II) obtained in the previous stage in 30 ml of dry THF is
introduced at this temperature and in 10 min. After 30 min at
0.degree.C., the mixture is brought to reflux for 10 min and then
cooled rapidly to 0.degree.C. 4.6 ml of 15% (w/v) NaOH solution and
then 4.6 ml of water are then added dropwise with caution. After 30
min contact, the mixture is filtered on a buchner provided with a
bed of infusorial earth. The filtrate is concentrated in vacuo and
on a water bath to obtain
(-)E-2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperidine(I) in
the form of a yellow oil which is verified pure by TLC. Weight: 3.2
g-yield: 84.6%-[.alpha.].sub.D.sup.20=67.20 (MeOH, c=8.0)
[0047] TLC(CH.sub.2Cl.sub.2/MeOH 10% ammoniacal 90/10 v/v):
Rf=0.65-0.80. 1H-NMR (CDI.sub.3- TMS): .delta. (ppm) 0.00-0.20 (m,
2H) ; 0.40-0.70 (m, 2H); 0.70-1.10 (m, 1H); 1.10-1.80 (m, 6H);
2.10-2.80 (m, 6H); 2.90-3.20 (m, 1H); 6.20-6.40 (m, 2H); 7.10-7.50
(m, 3H).
[0048] optical purity>98%, determined by HPLC column
4.6.times.150 mm Chiracel ODH 5 .mu.m (Daicel) , elution with
hexane containing 0.1% of triethylamine, t.degree.=30.degree.C.
[0049] Hydrochloride: The base is dissolved in 60 ml of
dichloromethane, 7.0 ml of 5.5 N hydrochloric ether are added, and
then the solvents are removed by distillation. The solid residue is
crystallized by dissolution in an isopropanol/diethyl ether
mixture. The white insoluble matter is filtered and dried in vacuo
to constant weight. Weight: 5.40 g-yield=68%.
[0050] m.p.=182-183.degree.C.-[.alpha.].sub.D.sup.20=-19.200
(c=0.5; CH.sub.2Cl.sub.2)
[0051] Analysis (C.sub.18 H.sub.24 Cl.sub.3 N): % C, H, Cl, N in
accordance;
[0052] IR (KEir): 3400, 2900, 2500, 1450, 1130, 1020, 990, 820, 800
cm.sup.-1
[0053] According to this process, starting from the distomeric salt
removed in the resolution in stage a), recovery and purification
allow the (-) antipode corresponding to the intermediate compound
(III) to be recovered, from which the distomer of the compound of
the invention is prepared, namely (+)E
2-(3,4-dichlorocinnamyl)-1-cyclopropylmethylpiperid- ine,
synthesized as comparison product.
EXAMPLE 2
[0054] Toxicity Study
[0055] In animals, the toxicity study was carried out in mice by
the oral route. No mortality was noted up to the strongest dose
studied, namely 300 mg/kg. No toxic symptoms were evident at doses
lower than 100 mg/kg. Starting from this dose, symptoms were
observed such as respiratory depression, trembling, convulsions, a
state of prostration or of lethargy which were reversible in a
lapse of time of 24 hours after treatment.
EXAMPLE 3
[0056] In vitro Pharmacological Study: Affinity for Sigma
Receptors
[0057] The study of the capacity of interaction with the sigma
receptors of the eutomer which is the subject of the invention, of
its antipode and of its parent racemic compound, was carried out by
the determination of their binding to a rat brain membrane
preparation, previously loaded with a labelled ligand specific for
sigma receptors, in this; case (+)[3H]-SKF 10,047. The technique
used (described by Largent, B. L. et al., J. Pharmacol. Exp. Ther.,
1986, vol. 238, pp. 739-748) consists in incubating, in solutions
of variable concentrations of the test product, the membrane
preparation previously loaded with (+)[3H]-SKF 10,047. After
filtration, the radioactivity of the solution, which is
representative of the displacement of the ligand by the test:
product, is measured. Results are expressed as ICSO of the test
product , which is the concentration allowing the tritiated ligand
to be displaced from 50% of its binding sites in the membrane
preparation
[0058] Results:: The values obtained are presented in Table 1:
1TABLE 1 In vitro study - .sigma. affinity .sigma. affinity: Test
compound IC.sub.50 (nM) (-)E 2-(3,4-dichlorocinnamyl)-1- 17.9
cyclopropylmethylpiperidine- (I) (eutomer) (+)E
2-(3,4-dichlorocinnamyl)-1- 21.8 cyclopropylmethylpiperidine
(antipode) (+/-)E 2-(3,4-dichlorocinnamyl)-1- 44.1
cyclopropylmethylpiperidine (racemate)
EXAMPLE 4
[0059] In vivo Pharmacological Studies
[0060] 4-1)--inhibition of experimental toxogenic diarrhoeas
Methods
[0061] a) diarrhoea induced by Salmonella lipoiolysaccharide
(LPS):
[0062] The test is carried out in mice according to a working
procedure, instigated by M. J. CANCIO et al., Gastroenterology
November 1992, 103 (5), 1437-43, which in the rat induces, by an
endotoxin, alterations in the transport of water and of
electrolytes at the colon level.
[0063] working procedure: dBA.sub.2 male mice (Iffa-credo, les
Oncins, France) weighing between 20 and 25 g are placed in
individual cages. After adaptation of the animals to their
environment, the test product is administered by the oral route in
solution or in aqueous suspension and then after one hour (to of
the test) an injection of lipopolysaccharides (LPS) of Salmonella
enteriditis (Sigma ref. L6761) is carried out in the tail vein at a
rate of 15 mg/kg. A preweighed filter paper is then placed under
each cage and the weight of faeces eliminated by the animals in two
hours (t.sub.120) is determined. The effect of the test product is
determined and is expressed as a percentage of inhibition of the
weight of faeces at the dose considered with respect to the weight
of faeces of a batch of control animals only having received LPS
under the same conditions. These results allow the ED.sub.50 of the
compounds to be calculated, which is the effective dose allowing
50% of the weight of the faeces caused by the administration of the
diarrhoeal agent to be inhibited.
[0064] b)--diarrhoea induced by the thermostable toxin of E. coli:
Nourished male NMRI mice (30-35 g) are weighed and placed in
individual cages previously covered with white paper allowing the
viewing of the faecal matter voided. The faeces are recovered as
soon as voided and are brought together, by periods of 30 min, for
120 min. The faeces thus brought together are weighed before (fresh
weight) and after (dry weight) drying at 120.degree.C. for 24 h.
The quantity of water present in the faeces is calculated as the
difference (fresh weight--dry weight). The thermostable toxin of
Escherichia coli (Sigma, E5763) is administered by the oral route
at time zero at a dose of 600 U/mouse. The animals of the control
batch receive, at time zero, an oral administration of saline. The
administration of the test products is carried out by the oral
route 1 h before the administration of the toxins. The results are
expressed as the quantity of faecal water amassed at time 120 min,
and allow the ED.sub.50 of the compounds to be calculated, which is
the effective dose allowing 50% of weight of water of the faeces to
be inhibited under the action of the test product.
[0065] c)--diarrhoea induced by the A and B toxins of C. difficile:
the experimental protocol, the calculation and the expression of
the results are identical to those described above. The A and B
toxins of C. difficile are administered by the oral route at time
zero at a dose of 6 ng/mouse.
[0066] Results
[0067] i)--Compared inhibitory activity of the levorotatory eutomer
(I) with the (+/-) parent racemate in mice, p.o administration
2TABLE 2 Inhibitory activity ED.sub.50 (mg/kg) Toxigenic
Laevorotatory (+/-) parent diarrhoea eutomer (I) racemate
Salmonella LPS 0.0056* 0.038 E. coil 0.019 0.077 C. difficile
<0.001 0.058 *inhibitory effect not reversed by naloxone
[0068] ii)--compared inhibitory activity of the levorotatory
eutomer (I) with loperamide (INN), igmesine (INN) and acetorphan
(INN) in mice, p.o administration
3TABLE 3 Inhibitory activity: ED.sub.50 (mg/kg) Toxigenic eutomer
diarrhoea (I) loperamide igmesine acetorphan Salmonella LPS 0.0056
0.37 0.42 34 E. coli 0.019 3.92 1.72 N.T. C. difficile <0.001
4.27 1.45 N.T. N.T.: not tested
[0069] iii)--diarrhoea induced by Salmonella LPS: compared
inhibitory activity of the levorotatory eutomer (I) with
(-)E-2-cinnamyl-1-cycloprop- ylmethylpyrrolidine (Ex. 1.3 of WO
95/15948) and (-)E-2-cinnamyl-1-cyclopr- opylmethyl-piperidine
(Example 2A.3) in mice, p.o. administration.
4TABLE 4 Inhibition of diarrhoea (%) Dose Test product administered
Levorotatory (mg/kg) eutomer (I) Ex. 1.3 Ex. 2A.3 0.00001 29
0.00003 45 0.0001 65.8 0.001 7.1 27.9 36.8 0.003 46.1 63.8 0.01
59.7 78.8 29.5 0.03 59.5 46.7 0.1 69.5 12.7 ED.sub.50 0.0056 0.002
0.00004 (mg/kg)
[0070] These results demonstrate the therapeutic advantage provided
by the compound of the invention, as it shows a significant effect
at a dose of 0.003 mg/kg (46.1%), an effect which continues up to a
dose of 0.1 mg/kg and beyond. Differently, the comparison compounds
only manifest a worthwhile activity within narrow dose limits:
[0071] from 0.003 to 0.03 mg/kg (63.8 to 46.7%) for the compound of
Example 1.3,
[0072] from 0.00003 to 0.001 mg/kg (45 to 36.8%) for the compound
of Example 2A.3, the activity decreasing considerably at the higher
doses for these two compounds.
[0073] 4-2)--inhibition of intestinal secretion induced by cholera
toxin: so-called enteropoolinq technique.
[0074] Female Wistar rats (160-180 g) are made to fast from solid
food 24 h before the test. At time zero, the animals receive, by
the oral route, 0.1 mg/kg of cholera toxin (Sigma, C3012). Three
hours afterwards, the animals are sacrificed by cervical
dislocation. After median laparotomy, the intestine is ligated at
the level of the pylorus and the ileocaecal junction. It is then
removed (from the duodenum to the caecum), and weighed full and
them empty. The administration of the test products is carried out
by the oral route one hour before the administration of the cholera
toxin. The results are expressed by weight of intestinal contents
and allow the ED.sub.50 of the compounds to be calculated, which is
the effective dose allowing the weight of the intestinal contents
to be inhibited by 50% under the action of the test product.
[0075] Results
5TABLE 5 Inhibition of intestinal secretion induced by cholera
toxin: ED.sub.50 (mg/kg) Laboratory eutomer (I) (+/-) parent
racemate 0.038 1.9
EXAMPLE 5
[0076] Pharmaceutical Forms
[0077] By way of illustration, the formulation and the preparation
of the hydrochloride of the levorotatory eutomer (I) of the
invention in the form of capsules and of tablets containing 40 mg
of active principle per unit is presented.
[0078] 5- 1 - Unit formulation of capsules and preparation:
6 Hydrochloride of the eutomer (I) 40.0 mg Lactose 59.3 mg
Colloidal silica 0.2 mg Magnesium stearate 0.5 mg Total 100 mg
[0079] The powders are intimately mixed and then distributed at a
rate of 100 mg per unit into capsules of appropriate size.
[0080] 5 - 2 - Unit formulation of tablets and preparation:
7 Hydrochloride of the eutomer (I) 40.0 mg Lactose 72.0 mg
Hydroxypropylmethylcellulose (HPMC) 3.6 mg Crosslinked
carboxymethylcellulose 3.6 mg (CMC) Colloidal silica 0.24 mg
Magnesium stearate 0.6 mg Total 120 mg
[0081] The active principle is mixed into the lactose and then
granulated with the HPMC in solution. The grains are dried and
screened on a grid of 1 mm mesh. The CMC and the silica are mixed
and then added to the granules. The mixture is then mixed
intimately with the magnesium stearate and then compressed at a
rate of 120 mg per scored tablet.
* * * * *