Derivatives of the N .alpha.-arylsulphonylaminoacyl-p-amidinophenylalaninamides, their preparation process, their use as medicaments and the pharmaceutical compositions containing them

The present invention is concerned with compounds with the formula: ##STR1## in which: R.sub.1 represents a lower alkyl, a lower hydroxy alkyl or a benzyl group, a phenyl group or a 4-hydroxy phenyl group: PA0 R.sub.2 and R.sub.3, identical or different, each represents a lower alkyl or hydroxyalkyl, lower alkenyl, or lower alkynyl radical, or together with the nitrogen to which they are attached, they form a saturated heterocycle such as morpholino, thiomorpholino, pyrrolidino unsubstituted or substituted by an alkoxycarbonyl or carboxyl group, piperazino, 4-(lower alkyl)-piperazino, 4-(lower hydroxyalkyl)-piperazino, or piperidino unsubstituted or substituted by a lower alkyl, benzyl, hydroxy, lower hydroxyalkyl, amino, lower aminoalkyl, alkoxycarbonyl or carboxyl group, PA0 Ar represents a phenyl, an alpha-naphthyl or a possibly substituted beta-naphthyl group, or a possibly substituted heteroaryl group chosen from pyridyl, quinolinyl, and isoquinolinyl, as well as their stereoisomers and their mixtures and their salts with pharmaceutically acceptable mineral or organic acids. The invention is also concerned with a preparation process for the products with the formula (I), their use as medicaments and the intermediates for their synthesis.

The present invention is concerned with new derivatives of 
N.alpha.-arylsulphonylaminoacyl-p-amidino-phenylalaninamides, their 
preparation process and their use as selective inhibiting agents of 
thrombin and antithrombotics. 
The compounds of the invention answer to the general formula (I) 
##STR2## 
in which: 
R.sub.1 represents a lower alkyl, lower hydroxyalkyl, or benzyl group, a 
phenyl or a 4-hydroxyphenyl group. 
R.sub.2 and R.sub.3, identical or different, each represents a lower alkyl 
or hydroxyalkyl, lower alkenyl or lower alkynyl radical, or they form 
together with the nitrogen to which they are attached, a saturated 
heterocycle such as morpholino, thiomorpholino, pyrrolidino unsubstituted 
or substituted by an alkoxycarbonyl or carboxyl group, piperazino, 
4-(lower alkyl)-piperazino or piperidino unsubstituted or substituted by a 
lower alkyl, benzyl, hydroxy, lower hydroxyalkyl, amino, lower aminoalkyl, 
alkoxycarbonyl or carboxyl group. 
Ar represents a phenyl, a possibly substituted alpha-naphthyl or 
beta-naphthyl group, or else a heteroaryl group chosen from pyridyl, 
quinolinyl, isoquinolinyl, possibly substituted. The preferred compounds 
with the above formula (I) are those in which R.sub.1 represents an alkyl 
or hydroxyalkyl radical, those in which the group 
##STR3## 
represents a piperidino radical, substituted or not, and those in which Ar 
represents a naphthyl radical, substituted or unsubstituted. The carbon 
carrying the group R.sub.1 can have R or S or RS configuration. All the 
compounds presenting the said configurations are included in the present 
invention. The compounds with the above formula (I) including one or more 
asymmetric centres can exist in the form of several isomers 
(diastereoisomers, enantiomers) and crystallization can bring in an 
enrichment of certain of the diastereo-isomers. The invention is concerned 
equally with each stereo-isomer and with their mixtures. The invention is 
concerned also with the addition salts of the compounds with the formula 
(I) with pharmaceutically acceptable mineral or organic acids. The terms 
"lower alkyl", "lower alkenyl" and "lower alkynyl" as used here, designate 
the radicals of branched or linear aliphatic hydrocarbon radicals 
containing up to six carbon atoms, such as methyl, ethyl, isopropyl, 
isobutyl, tertbutyl, n-hexyl, allyl, propargyl, crotyl, 2-methyl crotyl, 
2-methyl allyl, 2-butyryl. Synthetic inhibitors of thrombin, having an 
amidino-phenylalanine group have been described in the literature. G. 
WAGNER and his collaborators (GD Pat. No. 142804 (16.07.80)) have 
described the compounds with the general formula (A): 
##STR4## 
The insertion of a glycine amino-acid residue between the sulphonyl group 
and the nitrogen N- of the p-amidinophenylalanine led to the compounds 
with the general formula (B), of which the activity in vitro is enhanced 
in comparison with those with the general formula (A) (G. WAGNER et Coll, 
GD Pat. No. 155954 (03.02.81)): 
##STR5## 
and, among the latter, the compound with the formula (B) where n=1, 
Ar'=beta-naphthyl, NR'.sub.1 R'.sub.2 =piperidino, hereafter designated 
compound (C) offers the best thrombin inhibiting activity in vitro (J. 
STURZEBECHER et al, Thrombosis Research 1983, 29, 635) and ex-vivo (J. 
HAUPTMANN et al, Thrombosis Research 1985, 39, 771). The products with the 
formulae (A) and (B) above are prepared according to the processes 
described in the patents GDR No. 142804 and GDR No. 155954, the amides are 
prepared starting from the corresponding free acids by activation and 
reaction with the corresponding amine. These processes imply reaction 
conditions which induce racemisations at the asymmetric centre. 
Furthermore, they do not permit compounds to be obtained carrying the 
substituent R.sub.1. 
The Applicant has found that the compounds with the formula (I) above can 
be obtained by a process which, by the use of carefully chosen coupling 
processes and protector groups, enables the asymmetric centres to be 
respected in their original configuration and does not induce 
racemisation. 
This result, obtained, in contrast to the processes described by G. WAGNER 
and his collaborators, by first constructing the amide part 
##STR6## 
starting from the acid function of the synthon p-cyanophenylalanine, 
before the arylsulphonylaminoacyl part. The invention also has as its 
subject a preparation process for the compounds with the formula (I), 
characterized in that on the 4-cyanophenylalaninamide with the formula 
(II): 
##STR7## 
in which R.sub.2 and R.sub.3 have the same significances as in the formula 
(I), an acid with the formula 
##STR8## 
in its activated form (IV) 
##STR9## 
in which Ar and R.sub.1 have the same significances as in the formula (I) 
and R represents a good nucleofuge group, such as chloro, 
alkoxycarbonyloxy or heteroaryl, is made to react, so as to obtain the 
compound with the formula (V) 
##STR10## 
in which Ar, R.sub.1, R.sub.2 and R.sub.3 have the same significances as 
in formula (I), which is treated with an excess of a saturated solution of 
hydrogen chloride gas in an alcohol with the formula X--OH in which X 
represents a lower alkyl radical, so as to obtain the compound with the 
formula (VI) in the hydrochloride form: 
##STR11## 
in which Ar, R.sub.1, R.sub.2, R.sub.3 and X have the same significances 
as those already stated. 
The imido-ester of formula (VI) is then treated with an excess of an 
ammonia gas solution in a lower alcohol at the boiling temperature of the 
reactional mixture in order to obtain the compound sought with the formula 
(I). This compound is isolated in the form of a salt, it being possible to 
obtain the free base by standard processes and to convert it into another 
pharmaceutically acceptable salt such as, for example, in addition to the 
hydrochloride, hydrobromide, sulphate, methanesulphonate, acetate, 
2-naphthalene-sulphonate, maleate, fumarate, citrate, gluconate, 
dobesilate or sultosilate. 
The preparation of the new compound with the formula (II) will be described 
below and is carried out starting with the 4-cyanophenylalanine with the 
formula: 
##STR12## 
The acid with the formula 
##STR13## 
has been prepared by the following reactional scheme: 
##STR14## 
The introduction of an asymmetric centre in the amino-ester (VII: A=lower 
alkyl) of which the initial "R" or "S" configuration has to be preserved 
until the acid (III), requires the use of non-racemising methods. 
the sulphonylation of the amino-ester (VII) takes place in a bi-phase 
medium, preferably a water/dichloromethane, water/chloroform, or 
water/carbon-tetrachloride mixture, in the presence of a base, preferably 
an alkaline carbonate such as the carbonate of potassium or of sodium, at 
temperatures between 10.degree. C. and 25.degree. C. 
The saponification of the ester (VIII) takes place in a water/alcohol 
medium, such as water/methanol or water/ethanol, in the presence of an 
equivalent of an alkaline hydroxide, preferably sodium hydroxide, at 
temperatures between 10.degree. C. and 25.degree. C. The neutralization of 
the reactional medium by the addition of an equivalent of a 1N aqueous 
solution of a mineral acid, preferably hydrochloric acid, leads to the 
acid (III). This saponification can also be brought about in a 
water/organic medium, such as water/dioxan, in the same conditions. If it 
is not necessary to preserve the configuration of the asymmetric carbon, a 
standard method can be used. For the conversion of the acid with the 
formula (III) into an activated form with the formula (IV), two cases are 
to be considered: 
(a) Case where the carbon carrying the group R.sub.1 has the configuration 
"RS" (racemic): (non-stereo-specific synthesis). 
The activation of the acid function can be done, for example, either by: 
conversion of the acid function into an acyl halogenide (IV: R=Cl): 
##STR15## 
according to the process described in the GDR Pat. No. 155954, or by: 
conversion of the acid function into a mixed carbonic anhydride 
##STR16## 
according to the reactional scheme: The reaction uses an alkyl 
chloroformate 
##STR17## 
where Y.sub.1 is a lower alkyl radical, branched or not, in the presence 
of a tertiary amine as base. The alkyl chloroformate preferentially used 
is ethyl chloroformate, (Y.sub.1 =C.sub.2 H.sub.5) or isobutyl 
chloroformate, (Y.sub.1 =CH.sub.2 --CH(CH.sub.3).sub.2). 
The tertiary amine preferred is triethylamine. This condensation preferably 
takes place at temperatures between -5.degree. C. and +10.degree. C., in 
an inert solvent such as dichloromethane, chloroform or carbon 
tetrachloride. 
The compounds obtained with the formula (IV) are made to react with the 
compounds with the formula (II) in an inert solvent, in the presence of an 
acid acceptor, such as a tertiary amine. 
(b) Where the carbon carrying the group R.sub.1 has the configuration "R" 
or "S" (enantiomers R or S) (Stereo-specific synthesis). 
In this case it is necessary to use processes which do not induce 
racemisation. 
The conversion of the acid function of the compounds with the formula (III) 
into activated esters leads to compounds with the general formula (IV: 
R=O--Z) according to the reactional scheme: 
##STR18## 
The coupling reagents Y.sub.2 --Z, which do not induce racemization, used 
for preference but which are not limitative, are the following: 
1-hydroxybenzotriazole (HOBT) 
##STR19## 
in the presence of N,N-dicyclohexylcarbodiimide (DCC) according to the 
method of operation described by E. C. JORGENSEN et al (J. Am. Chem. Soc. 
1971, 93, 6318) 
1-benzotriazolyloxy-tris(dimethylamino)phosphonium hexafluorophosphate 
(BOP) 
##STR20## 
according to the operational method described by B. CASTRO et Coll. 
Synthesis 1976, 751. 
N,N-bis(2-oxo-3-oxazolidinyl)phosphorodiamide chloride 
##STR21## 
according to the operational method described by D. H. RICH et al (J. Am. 
Chem. Soc. 1985, 107, 4342. 
The activation and coupling reaction takes place in the presence of 
tertiary amines, preferably triethylamine, in an inert solvent such as 
dichloromethane, dimethylformamide or acetonitrile, at temperatures 
between 15.degree. C. and 40.degree. C. 
The formation of the imido-ester (VI) takes place in an alcohol medium such 
as methanol or ethanol, at a temperature between -10.degree. C. and 
+10.degree. C., preferably at 0.degree. C., during a period of 16 to 24 
hours. 
The amidine of formula (I) is obtained by treating the compound (VI) 
previously obtained, without further purification, in an alcohol solution 
of ammonia gas at a normality of 3N to 15N, at ambient temperature, and 
the mixture is then heated to reflux for 1 to 3 hours. 
The compounds with the formula (II) above, including one asymmetric center 
can exist in the form of two isomers (enantiomers). The invention is 
concerned with each stereoisomer as well as their mixtures. The invention 
is also concerned with their addition salts with mineral or organic acids. 
The present invention is also concerned with a process for preparing the 
compounds with the formula (II), wherein an amino acid with the formula 
(IX) in its activated form (X): 
##STR22## 
in which R' represents an N-protecting group and A represents the moiety 
of a coupling reagent, are reacted with an amine having the formula (XI): 
##STR23## 
in which R.sub.2 and R.sub.3 are as in formula (II), in order to form a 
compound with the formula (XII): 
##STR24## 
in which R', R.sub.2 and R.sub.3 have the same significances as in 
formulae (II) and (IX), and which, by cleavage of the protecting group R' 
gives the compounds of formula (II). 
The formation of the compound with formula (IX) is obtained by fixing 
N-protecting group R' on p-cyanophenylalanine with the formula: 
##STR25## 
The N-protecting group, represented by R', is a group stable in alkaline 
medium used for the protection of the amino group of aminoacids in the 
peptide chemistry, for example the tert.butyloxycarbonyl group, preferably 
designed hereafter Boc; the 2-(3,5-dimethoxyphenyl) 2-propyl-oxycarbonyl 
group designed by Ddz; the 2-(4-biphenyl) 2-propyl-oxycarbonyl group 
designed by Bpoc; the (2-nitrophenyl)-sulphenyl group designed by Nps. 
In order to obtain the activated acid with the formula (X) in which A 
represents the coupling reagent moiety, two cases are to be considered. 
(a) Preparation process with conservation of the "R" or "S" configuration 
(stereo-specific synthesis) 
In order to avoid inducing racemisation at the asymmetric center of the 
compound with the formula (XII) and to keep the initial configuration of 
the asymmetric center of the acid with the formula (IX), it is necessary 
to carry out the activation of the acid (IX) using the transformation of 
the acid into an activated ester (X) according to the reaction schema: 
##STR26## 
The coupling reagents Y--Z, which do not induce racemisation, preferably 
used, but not limitative, are the following: 
1-Hydroxybenzotriazole (HOBT) 
##STR27## 
in the presence of N,N-dicyclohexylcarbodiimide (DCC) according to the 
method described by E. C. JORGENSEN et al. (J. Am. Chem. Soc. 1971, 93, 
6318). 
1-Benzotriazolyl-oxytris(dimethylamino)phosphonium hexafluorophosphate 
(BOP) 
##STR28## 
according to B. CASTRO et al. (Synthesis 1976, 751). 
N,N-bis(2-oxo-3-oxazolidinyl)phosphorodiamide chloride 
##STR29## 
according to the method described by D. H. RICH et al. (J. Am. Chem. Soc. 
1985, 107, 4342). 
The activation and coupling reaction is carried out in the presence of 
tertiary amines, preferably triethylamine, in an inert solvent such as 
dichloromethane, dimethylformamide or acetonitrile, at temperatures 
comprised between 15.degree. C. and 40.degree. C. 
(b) Preparation process without conservation of the configuration 
(non-stereo-synthesis) 
The action of the acid function of the compound (IX) may be carried out by 
transformation of the acid function into a mixed anhydride (X: 
A=--O--CO--O--B) according to the reaction schema 
##STR30## 
The reaction uses an alkyl chloroformate 
##STR31## 
wherein B is a lower linear or non linear alkyl, in the presence of a 
tertiary amine as base. The alkyl chloroformate preferably used is ethyl 
(B=C.sub.2 H.sub.5) or isobutyl (B=CH.sub.2 --CH(CH.sub.3).sub.2) 
chloroformate. The preferred tertiary amine is triethylamine. This 
condensation is preferably carried out at temperatures comprised between 
-5.degree. C. and +10.degree. C., in an inert solvent such as 
dichloromethane, chloroform or carbon tetrachloride. 
The starting product p-cyanophenylalamine was prepared according to one of 
the methods used in the literature (G. WAGNER et al. Pharmazie 1981, 36 
(9), 597). 
The compound with the formula (X) is reacted with the amine of formula (XI) 
in an inert solvent and in the presence of a tertiary amine. 
The cleavage of the N-protecting group R' of the compound with the formula 
(XII) leads to N.alpha.-substituted p-cyanophenylalamines with the formula 
(II). This cleavage is carried out in acidic medium, preferably a 
hydrobromic acid-acetic acid mixture or in trifluoroacetic acid (R'=Boc, 
Ddz, Bpox, Nps), in acetic acid (R'=Boc, Nps), in an ethyl acetate R'=Boc) 
solution saturated with hydrochloric gas, at temperatures comprised 
between 0.degree. and 20.degree. C.

EXAMPLE 1 
N.alpha.-(tertubyloxycarbonyl)p-cyanophenylalanine 
##STR32## 
10 g (0.044 mole) of p-cyanophenylalanine and 88.2 ml (0.088 mole) aqueous 
1N sodium hydroxyde are dissolved in 220 ml of dioxane. At ambient 
temperature and under an inert atmosphere, 1.77 g (0,044 mole) of 
magnesium and then 10.6 g (0.0484 mole) of diterbutyl dicarbonate are 
progressively added to the reaction medium. The medium is stirred at 
ambient temperature for 20 hrs. The crystals are filtered off and washed 
with water. The filtrate is evaporated and the residue is dissolved in 
water. The aqueous phase obtained is brought to pH=3 by adding a solution 
saturated with potassium hydrogenosulphate. The aqueous phase is extracted 
with 2.times.400 ml ethyl acetate, the organic extracts are dried on 
anhydrous sodium sulphate and evaporated to dryness. The crystals obtained 
are recrystallized in ethyl acetate or diisopropyl ether. 
White crystals, yield: 88%, M.P.=147.degree. C. 
EXAMPLE 2 
1-[N.alpha.-(terbutyloxycarbonyl)p-cyanophenylalanyl]piperidine 
##STR33## 
Activation of the acid function of the compound (IX: 
R=--C--OC(CH.sub.3).sub.3) (example 1) into mixed anhydride 
##STR34## 
4.2 g (0.0416 mole) of triethylamine is added to a suspension of 11 g 
(0.0378 mole) of N.alpha.-(terbutyloxycarbonyl)p-cyanophenylalanine (Ex. 
1) in 125 ml dichloromethane, at 0.degree. C., under an inert atmosphere. 
To the mixture which becomes homogeneous, a solution of 4.3 g (0.0395 
mole) ethyl chloroformate in 10 ml dichloromethane is added dropwise. At 
the end of the addition, the reaction mixture is allowed to stand for 45 
minutes at 0.degree. C., then 3.4 g (0.0397 mole) piperidine dissolved in 
10 ml dichloromethane is added dropwise at 0.degree. C. The reaction 
mixture is allowed to return to ambient temperature and is left at this 
temperature for 15 hrs. The reaction medium is extracted with an aqueous 
solution saturated with sodium bicarbonate. The organic phase, after 
decantation, is dried on anhydrous sodium sulphate and evaporated to 
dryness. The oily residue, after trituration with diisopropyl ether, gives 
white crystals. These crystals are recrystallized in diisopropyl ether. 
White crystals, yield: 81%, M.P. =132.degree. C. 
EXAMPLE 3 
4-methyl 1-[N.alpha.-(terbutyloxycarbonyl)p-cyanophenylalanyl]piperidine 
##STR35## 
Activation by transformation of the acid function into the activated ester 
function, by using the non-racemising coupling reagent DCC/HOBT. 
2.78 g (0.01 mole) of N.alpha.-(tertbutyloxycarbonyl)p-cyanophenyl-alanine 
(Ex. 1) is dissolved in 50 ml of dichloromethane. Under an inert 
atmosphere, at ambient temperature, there are added successively 1 g (0.01 
mole) of 4-methylpiperidine, 1.35 g (0.01 mole) of 1-hydroxy-benzotriazole 
(HOBT), 1.1 g (0.01 mole) of triethylamine. To the reaction medium there 
is added 2.06 g (0.01 mole) of N,N-dicyclohexylcarbodiimide (DCC) 
dissolved in 80 ml of dichloromethane at ambient temperature and the 
reaction medium is left for 15 hrs. at ambient temperature. The 
precipitate of dicyclohexylurea is filtered off and eliminated. The 
organic filtrate is washed with a saturated aqueous solution of sodium 
bicarbonate, and dried on anhydrous sodium sulphate. Evaporation gives a 
residue which is triturated with diisopropyl ether. The white crystals 
obtained are recrystallized in ethyl acetate. 
White crystals, yield: 84%, M.P.=142.degree. C. (ethyl acetate) 
EXAMPLE 4 
1-(p-cyanophenylalanyl)-piperidine (II: NR.sub.2 R.sub.3 
=4-methylpiperidino) 
4.1 g (0.0115 mole) of 
1-[N.alpha.-(terbutyloxycarbonyl)p-cyanophenylalanyl]piperidine (example 
1) dissolved in 22 ml of ethyl acetate is added to 22 ml of a solution of 
ethyl acetate saturated with hydrochloric gas, cooled to 0.degree. C. 
There is a progressive dissolution. The mixture is allowed to return to 
ambient temperature and left at ambient temperature for 15 hrs. The 
crystals precipitate, are filtered off, washed with ether and 
recrystallized in isopropanol. 
White crystals, yield: 88%; M.P.=145.degree. C. (hydrochloride). 
EXAMPLE 5 
1-(p-cyanophenylalanyl)-4-methyl-piperidine (II: NR.sub.2 R.sub.3 
=4-methyl-piperidino) 
Prepared according to the method described in Example 4, from 4-methyl 
1-[N.alpha.-(terbutyloxycarbonyl)p-cyanophenylalanyl]piperidine (Example 
3). 
White crystals, yield: 97%; M.P.=210.degree. (isopropanol), hydrochloride. 
EXAMPLE 6 
1-[N.alpha.-(N.alpha.-betanaphthylsulphonyl-(R)-phenylalanyl)-p-cyanophenyl 
alanyl]piperidine (V: Ar=betanaphthyl; R.sub.1 =CH.sub.2 --C.sub.6 H.sub.5 
; NR.sub.2 R.sub.3 =piperidino) 
To a suspension of 12.5 g (0.0245 mole) of 
1-(p-cyanophenylalanyl)piperidine hydrochloride in 200 ml of 
dichloromethane there are added successively 15.1 g (0.0245 mole) of 
N-(betanaphthylsulphonyl)-(R)-phenylalanine, 4.3 g (0.0425 mole) of 
triethylamine, 6.5 g (0.0425 mole) of 1-hydroxybenzotriazole (HOBT). The 
reactional medium is cooled to between 0.degree. C. and 5.degree. C., then 
8.8 g (0.0425 mole) of N,N-dicyclohexylcarbodiimide (DCC) dissolved in 50 
ml of dichloromethane is added drop by drop. The reactional medium is left 
under good agitation at ambient temperature for 17 hours. The precipitate 
of dicyclohexylurea is filtered off, and the organic filtrate is washed 
with a saturated aqueous solution of sodium bicarbonate. The organic 
phase, dried on anhydrous sodium sulphate, is evaporated to dryness. The 
evaporation of the solvent leaves a residue which is triturated with ethyl 
acetate. The white crystals are filtered off and washed wih diisopropyl 
ether. 
White crystals, yield: 64%, m.p.=173.degree. C. (ethyl acetate). 
Examples 7 to 12 use the same methods of operation as are described in 
example 6. They lead to the nitriles of formula (V) and result from the 
coupling of the synthons, with the general formula (II), with the acids 
with the general formula (III), previously activated by conversion of the 
acid function into an activated ester function (IV), using the 
non-racemising coupling reagent DCC/HOBT. They are summarized in the 
following table. 
______________________________________ 
##STR36## 
(V: Ar = betanaphthyl) 
R.sub.1 (configuration m.p. 
Example 
of the amino acid) 
NR.sub.2 R.sub.3 
Yield .degree.C. 
______________________________________ 
7 CH.sub.3 (R) 
##STR37## 51% 160.degree. 
8 C.sub.6 H.sub.5 (R) 
##STR38## 38% 194.degree. 
9 CH.sub.2 OH (S) 
##STR39## 75% 97.degree. 
10 CH.sub.3 (R) 
##STR40## 73% 160.degree. 
11 CH.sub.3 (S) 
##STR41## 72% 150.degree. 
12 CH(CH.sub. 3)OH (S) 
##STR42## 78% 106.degree. 
______________________________________ 
EXAMPLE 13 
1-[N.alpha.-(N.alpha.-betanaphthylsulphonyl-(S)-alanyl)-p-cyanophenylalanyl 
]piperidine: (V: Ar=betanaphthyl; R.sub.1 =CH.sub.3 ; NR.sub.2 R.sub.3 
=piperidino) 
To a solution of 7.5 g (0.027 mole) of 
N-(betanaphthylsulphonyl)-(S)-alanine (III: R.sub.1 =CH.sub.3, 
Ar=betanaphthylsulphonyl) of S configuration in 300 ml of acetonitrile, 
there are added successively 11.9 g (0.027 mole) of 
1-benzotriazolyl-trisoxy(dimethylamino)phosphonium hexafluorophosphate 
(BOP), 6.9 g (0.027 mole) of 1-(p-cyanophenylalanyl)piperidine, (II: 
NR.sub.2 R.sub.3 =piperidino), 2.75 g (0.027 mole) of triethylamine. The 
reactional medium is left under an inert atmosphere, under good agitation, 
at ambient temperature, for 20 hours. The reactional medium is diluted 
with ethyl acetate and washed successively with a saturated aqueous 
solution of sodium chloride, a 2N solution of hydrochloric acid, water, 
and with a saturated aqueous solution of sodium bicarbonate, then with 
water. The organic phase is dried on anhydrous sodium sulphate and 
evaporated to dryness. The residue is purified by chromatography on a 
silica column (elution: toluene-ethyl acetate, 1:1). White crystals are 
recovered which are dried. 
White crystals, yield: 45%; m.p. 155.degree. C. 
EXAMPLE 14 
1-[N.alpha.-(N.alpha.-betanaphthylsulphonyl-(RS)-phenylglycyl)-p-cyanopheny 
lalanyl]piperidine: (V: Ar=betanaphthyl; R.sub.1 =C.sub.6 H.sub.5 ; 
NR.sub.2 R.sub.3 =piperidino) 
Under an inert atmosphere, 3.4 g (0.01 mole) of 
N-betanaphthylsulphonylphenylglycine (III: R.sub.1 =C.sub.6 H.sub.5 ; 
Ar=betanaphthyl) racemic (RS) in 30 ml of thionyl chloride is taken to 
reflux for one hour. The reactional medium is evaporated to dryness and 
the oily residue is dissolved in 50 ml of dichloromethane. The solution of 
acid chloride (IV:R.sub.1 =C.sub.6 H.sub.5, Ar=betanaphthyl: R=Cl) in 
dichloromethane is added drop by drop under an inert atmosphere to a 
solution of 1.3 g (0.005 mole) of 1-(p-cyanophenylalanyl)piperidine (II: 
NR.sub.2 R.sub.3 =piperidino) and of 0.6 g (0.005 mole) of triethylamine 
in 20 ml of dichloromethane, which has previously been cooled to between 
0.degree. and +5.degree. C. The reactional medium is left at ambient 
temperature for 20 hours. The insoluble salts are filtered off and the 
filtrate is evaporated to dryness. The residue is taken up 1N hydrochloric 
acid and the aqueous acid phase obtained is extracted with 
dichloromethane. The organic extracts are dried on anhydrous sodium 
sulphate and evaporated to dryness. The residue obtained after evaporation 
is purified by chromatography on a silica column (elution: toluene-ethyl 
acetate 1:1). White crystals are obtained. 
White crystals, yield: 80%; m.p.=190.degree. C. 
EXAMPLE 15 
1-[N.alpha.-(N.alpha.-betanaphthylsulphonyl-(RS)-valyl)-p-cyanophenylalanyl 
]piperidine (V: Ar=betanaphthyl; R.sub.1 =CH(CH.sub.3).sub.2 ; NR.sub.1 
R.sub.2 =piperidino) 
To a suspension of 7 g (0.023 mole) of N-betanaphthylsulphonylvaline (III: 
R.sub.1 =CH(CH.sub.3).sub.2 ; Ar=betanaphthyl) racemic (RS) in 10 ml of 
dichloromethane, kept at between 0.degree. and +5.degree. C., 2.6 g (0.023 
mole) triethylamine is added, then 3.4 g (0.025 mole) of isobutyl 
chloroformate is added drop by drop, and the whole is left for one hour at 
this temperature. Then 6.2 g (0.024 mole) of 
1-(p-cyanophenylalanyl)piperidine (II: NR.sub.2 R.sub.3 =piperidino) 
dissolved in 50 ml of dichloromethane is added, and the reactional medium 
is left for 20 hours at ambient temperature. After evaporating to dryness, 
the residue is taken up with water. The aqueous phase is extracted with 
dichloromethane. The organic extracts are dried on anhydrous sodium 
sulphate and evaporated to dryness. The oily residue is purified by 
chromatography on a silica column (elution: toluene-ethyl acetate, 1:1). 
White crystals, yield: 78%; m.p. 178.degree. C. 
EXAMPLE 16 
1-[N.alpha.-(N.alpha.-betanaphthylsulphonyl-(R)-phenylglycyl)-p-amidinophen 
ylalanyl]piperidine: (I: Ar=betanaphthyl; R.sub.1 =C.sub.6 H.sub.5 ; 
NR.sub.2 R.sub.3 =piperidino) derivative No. 1 
(a) Formation of the imido-ester. 
Under an inert atmosphere, 100 ml of methanol is saturated with hydrogen 
chloride gas at 0.degree. C., and 8.1 g (0.0139 mole) of 
1-[N-(N-betanaphthylsulphonyl-(R)-phenylglycyl)-p-cyanophenylalanyl]piperi 
dine (example 8) is added all at once, and the whole is left at 0.degree. 
C. for 20 hours. After evaporating to dryness without heating the 
methanol, a white resin is obtained, consisting of the hydrochloride of 
the imido-ester with the general formula (VII: Ar=betanaphthyl: R.sub.1 
=C.sub.6 H.sub.5 : NR.sub.2 R.sub.3 =piperidino), which is used without 
further purification in the last stage. 
(b) Formation of the amidine 
Under an inert atmosphere, 100 ml of methanol is saturated with ammonia gas 
at temperatures between 0.degree. C. and +5.degree. C., and the white 
resin obtained in example 10a, after being dissolved in 20 ml of methanol, 
is added to this methanol ammonia solution. The reactional mixture is 
taken to reflux under an inert atmosphere, for 3 hours. After evaporating 
to dryness, the residue is taken up with an excess of 1N hydrochloric 
acid. The aqueous acid phase is extracted with dichloromethane. The 
organic phase is dried on anhydrous sodium sulphate and evaporated to 
dryness. The semi-crystalline residue, obtained after evaporation, is 
taken up with water. The insoluble crystals in the water are recovered, 
dried, then treated with ethyl acetate, and taken to reflux for 10 
minutes. The insoluble crystals in the boiling ethyl acetate are filtered 
off and dried. 
White crystals, hydrated hydrochloride, yield: 84%; m.p. 188.degree. C. 
The examples 17 to 23 use the same operational methods as those described 
in example 16. They lead to the 
N.alpha.-arylsulphonylaminoacyl-p-amidinophenylalaninamides with the 
general formula (I) and result from the conversion of the nitriles with 
the general formula (V) by the intermediary of the imido-esters with the 
general formula (VI). They are tabulated in the following table. 
__________________________________________________________________________ 
##STR43## 
R.sub.1 
example 
Derivative 
Ar (configuration of the amino acid) 
NR.sub.2 R.sub.3 
x YIELD m.p. 
__________________________________________________________________________ 
.degree.C. 
17 2 
##STR44## CH.sub.2C.sub.6 H.sub.5 (R) 
##STR45## 
1 83% 178.degree. 
18 3 
##STR46## CH.sub.3 (R) 
##STR47## 
1,5 
53% 170.degree. 
19 4 
##STR48## CH.sub.3 (S) 
##STR49## 
1 54% 176.degree. 
20 5 
##STR50## CH.sub.3 (R) 
##STR51## 
2 69% 177.degree. 
21 6 
##STR52## CH.sub.3 (S) 
##STR53## 
2 64% 176.degree. 
22 7 
##STR54## CH.sub.2 OH(S) 
##STR55## 
1 51% 162.degree. 
23 8 
##STR56## CH(CH.sub.3)CH(S) 
##STR57## 
0,5 
68% 184.degree. 
__________________________________________________________________________ 
The results of the toxicological and pharmacological studies which are 
reported below have made apparent the useful properties of the invention 
compounds. These are endowed with a very good thrombin-inhibiting 
activity, and possess, in addition, remarkable antithrombotic properties 
in vivo, which compounds with the formulae (A) and (B) and the compound 
(C) do not show. Compared with heparin, they have a much greater duration 
of action without inducing an increase in the bleeding time. The subject 
of the invention is therefore also a medicament presenting, in particular, 
anti-thrombotic properties, characterized in that it contains, as active 
principle, a compound with the formula (I) or an addition salt with a 
therapeutically acceptable mineral or organic acid. 
Toxicological study 
The invention compounds enjoy a good tolerance and a weak toxicity. Tests 
carried out on different animal species on the acute, sub-chronic and 
chronic toxicity have not brought to light any sort of local or general 
reaction, perturbation or anomaly in the biochemical, macro-scopical and 
microscopical examinations carried out throughout the tests. 
Pharmacological study 
In this study, the invention compounds have been compared with heparin and 
with [N-(N-betanaphthylsulphonylglycyl)-p-amidinophenylalanyl]piperidine, 
a compound of closely similar structure described as a powerful thrombin 
inhibitor (J. HAUPTMANN & COLL., THROMB., RES., 38 771-775, 1983) and 
which will be called derivative C. 
(1) Thrombin time. 
The coagulation time of citrated plasma in the presence of thrombin is 
measured ex vivo in the rat, according to the technique of BIGGS R. M., 
(Human blood coagulation, haemostasis and thrombosis: Oxford, Blackwell 
Scientific Publications, 1972). 
The samples were taken one hour after sub-cutaneous administration of the 
compound under test, by puncture of the abdominal aorta. The blood was 
collected on sodium citrate at 3.8% (1 volume for 9 volumes of blood). The 
plasma was obtained by centrifuging at 2600 g for 10 minutes. To 0.2 ml of 
the plasma, 0.2 ml of a thrombine solution was added, (20 U/ml). The 
coagulation time was recorded. The results are assembled in the following 
table and expressed in seconds. 
______________________________________ 
Dose Result % ex- 
Product mg/Kg Route (time in secs.) 
extension 
p 
______________________________________ 
control s.c. 6 .+-. 0 
heparin 10 s.c. 20 .+-. 4 
230 0.001 
control s.c. 7 .+-. 0 
derivative C 
10 s.c. 9 .+-. 0 29 0.001 
control s.c. 5.9 .+-. 0 
derivative No. 7 
10 s.c. 48.6 .+-. 10.8 
782 0.001 
control s.c. 8.2 .+-. 0.2 
derivative No. 4 
10 s.c. 49.9 .+-. 6.2 
508 0.001 
control s.c. 7.7 .+-. 0.1 
derivative No. 6 
10 s.c. 12.7 .+-. 0.3 
65 0.001 
control s.c. 7 .+-. 0 
derivative No. 8 
10 s.c. 87 .+-. 11 
1104 0.01 
______________________________________ 
(2) Venous thrombosis with the spiral 
The tests were carried out according to an adaptation of the method of T. 
KUMADA et al., (Thromb. Res., 18, 189-203, 1980) A metallic spiral (re-cut 
dentist's paste rammer) is inserted in the inferior vena cava of an 
anesthetized rat. The animals had received the compound under test one 
hour earlier by sub-cutaneous route. Five hours later, the spiral was 
withdrawn with the thrombus which it retained, and was then dried by 
repeated dabbing with filter paper and weighed. The spiral was then 
relieved of the thrombus, dried and weighed again. The weight difference 
gives the weight of the thrombus. The results are assembled in the 
following table. 
______________________________________ 
Dose Weight of thrombus 
Product mg/Kg in mg Variation 
p 
______________________________________ 
control 4.47 .+-. 0.51 
heparin 5 2.91 .+-. 0.53 
-35% 0.05 
heparin 10 1.62 .+-. 0.34 
-64% 0.001 
heparin 20 0.26 .+-. 0.04 
-94% 0.001 
control 4.77 .+-. 0.47 
derivative C 
20 3.99 .+-. 0.45 
-15% n.s. 
derivative C 
50 3.63 .+-. 0.37 
-24% n.s. 
derivative C 
100 3.08 .+-. 0.28 
-35% 0.01 
control 3.81 .+-. 0.38 
derivative No. 4 
5 2.67 .+-. 0.22 
-30% 0.05 
derivative No. 4 
10 2.48 .+-. 0.35 
-35% 0.05 
derivative No. 4 
20 1.92 .+-. 0.14 
-50% 0.001 
control 3.72 .+-. 0.33 
derivative No. 4 
50 2.62 .+-. 0.26 
-30% 0.05 
control 3.72 .+-. 0.33 
derivative No. 7 
10 2.42 .+-. 0.26 
-35% 0.01 
control 4.28 .+-. 0.53 
derivative No. 8 
10 2.07 .+-. 0.12 
-52% 0.01 
______________________________________ 
(3) Bleeding time 
This study was carried out according to an adaptation of the technique of 
L. STELLA et al., (Thromb. Res.; 1975 7, 709-716). After anesthetizing a 
rat with pentobarbital, its tail was cut at 5 mm from the end and the 
blood from the wound was carefully dabbed every 15 seconds with a filter 
paper until haemostasia occurred. The latter was obtained when no stain 
appeared during one minute. The products to be tested are administered by 
sub-cutaneous route one hour before the tail was cut. The results are 
assembled in the following table: 
______________________________________ 
Dose Duration 
Product mg/Kg in seconds 
limits p 
______________________________________ 
control 360 330-480 
heparin 5 465 330-540 n.s. 
heparin 10 3600 525-&gt;3600 
0.01 
heparin 20 3600 690-&gt;3600 
0.01 
control 540 450-675 
derivative C 
10 375 360-510 n.s. 
derivative C 
20 750 420-960 n.s. 
derivative C 
50 600 435-615 n.s. 
control 405 390-510 
derivative No. 4 
10 440 400-520 n.s. 
derivative No. 4 
20 450 420-530 n.s. 
derivative No. 4 
50 480 420-3500 n.s. 
control 540 400-680 
derivative No. 6 
10 380 340-480 n.s. 
derivative No. 6 
20 550 420-700 n.s. 
derivative No. 6 
50 600 430-605 n.s. 
control 525 450-610 
derivative No. 7 
10 605 530-760 n.s. 
derivative No. 7 
20 585 495-860 n.s. 
derivative No. 7 
50 540 460-650 n.s. 
______________________________________ 
Bleeding time: this study has clearly shown the risk of hemorrhage induced 
by heparin which extends considerably the bleeding time. Being without 
action on the bleeding time, the invention derivatives possess a much 
greater safety margin than heparin does. 
The medicament of the invention can be presented for oral administration in 
the form of tablets, sugar-coated tablets, capsules, drops, syrup or 
granules. It can also be presented for rectal administration in the form 
of suppositories and for parenteral administration in the form of an 
injectable soluteion. Each unit dose advantageously contains from 0.005 g 
to 0.500 g of active principle as a function of the age of the invalid and 
of the seriousness of the affection treated. By way of non-limitative 
examples, there are given below several pharmaceutical formulations of the 
invention medicament. 
______________________________________ 
(1) tablets 
derivative No. 4 
0.050 g 
excipient lactose, corn starch, talc, 
colloidal silica, magnesium stearate. 
(2) sugar-coated tablets 
derivative No. 6 
0.025 g 
excipient sugar, corn starch, polyvinyl pyrro- 
lidone, silicic acid, magnesium 
stearate, talc, "orange yellow". 
(3) capsules 
derivative No. 3 
0.100 g 
excipient talc, wheat starch, magnesium 
stearate 
(4) suppositories 
derivative No. 5 
0.050 g 
excipient semi-synthetic glycerides 
(5) injectable solution 
derivative No. 11 
0.025 g 
excipient isotonic solvent, q.s. for 5 ml. 
______________________________________ 
For its anti-coagulant and anti-thromobotic properties, without secondary 
effects due to the risk of hemorrhagia, the invention medicament is 
usefully administered in the prevention and treatment of thrombo-embolic 
disease.