FR2730488A1 - New 15-deoxy:spergualine analogues - Google Patents

Abstract

15-Deoxyspergnaline (15-DSG) analogues of formula (I) and their addn. salts are new. In (I), Q is a gp. of formula (i); R is H, OH, OMe or CH2OH; asterisk C is an asymmetric carbon of configuration (R,S), (R) or (S), with the proviso that R is not H; double asterisk C is an asymmetric carbon of configuration (R,S) or (R). Also claimed are (A) intermediate cpds. of formula (II) which are useful in prepn. of (I) and/or the isomers 11-(5)-15-DSG and 11-(R)-15-DSG where Ar is an aromatic substit.; Ra is 1-3C alkoxy or HN-(CH2)4-N(R2)-(CH2)2-CH(Me)-NHR3; R1-R3 are amino-protecting gp.; and asterisk C is a carbon atom of configuration (S) or (R).

Description

Analogues of lS-deoxyspergualine, their use in therapeutics and
their preparation process
Field of the invention
The present invention relates as novel industrial products to 15-deoxyspergualine-related structure compounds.

It also relates to their method of preparation and their use in therapy.

Prior art
It is known that 15-deoxyspergualine (DSG), also known under the international nonproprietary name "Gusperimus", has an interesting activity in the field of immunosuppression. Numerous publications report this activity: a series of articles on this topic can be found in: "Immunomodulating Drugs" - Annals of the
New York Academy of Sciences, Vol 685, pages 123-201.

 However, 15-deoxyspergualin does not exhibit satisfactory chemical stability and more stable compounds have been sought, for example (i) by replacing the α-hydroxyglycine group of the deoxyspergualin with various α or α-amino acids, (ii) by modifying the structure of the central link, or (iii) by modifying the link carrying the guanidine function. Examples of such modifications are described in EP-A-0181 592, EP-A-0 105 193 and FR-A-2 698 628.

 The modifications relating to the spermidine chain were mainly studied in J. Antibiot. 40 pages 1303-1315 and most of the prepared compounds were inactive. None of the proposed structures showed an activity at least equivalent to that of the DSG and the authors concluded the essential presence of spermidine sequencing.

 The present invention relates to compounds analogous to 15-oxyspergualine, but whose spermidine chain has been modified and which have a higher activity than the known products.

Object of the invention
The present invention provides novel compounds, the general structure of which remains related to 15-deoxyspergualine, which have an activity superior to the products known from the prior art in the field of immunosuppression.

 The products according to the invention are distinguished in particular from the products known from the prior art by the presence of a methyl group on the carbon atom bearing the primary amine function of the spermidine chain of the molecule. The choice of a particular configuration of this asymmetric carbon atom (denoted hereinafter C) also makes it possible to improve the activity of these new compounds.

dans laquelle
- R représente un atome d'hydrogène, un groupe OH, un groupe OCH3
ou un groupe CH2OH,
- *C, dans le cas où R n'est pas un atome d'hydrogène, est un atome de
carbone asymétrique dont la configuration peut être indéterminée (R,S),
ou déterminée (R) ou (S),
- **C est un atome de carbone asymétrique de configuration
indéterminée (R,S) ou de configuration déterminée (R) ; et, (ii) leurs sels d'addition.The analogous compounds of 1'15-deoxyspergualine according to the invention are characterized in that they are chosen from the group consisting of: (i) compounds of formula

in which
R represents a hydrogen atom, an OH group or an OCH3 group;
or a CH2OH group,
- * C, in the case where R is not a hydrogen atom, is an atom of
asymmetric carbon whose configuration can be indeterminate (R, S),
or determined (R) or (S),
- ** C is an asymmetric carbon atom of configuration
indeterminate (R, S) or determined configuration (R); and, (ii) their addition salts.

dans laquelle:
- R représente un atome d'hydrogène, un groupe OCH3, un groupe OH,
un groupe CH2OH, un groupe OR' ou un groupe CH2OR',
- R' représente un groupe protecteur de la fonction hydroxyle,
- R1, R2, R3, identiques ou différents, représentent chacun un groupe
protecteur de la fonction amine,
- *C représente, lorsque R n'est pas l'atome d'hydrogène, un atome de
carbone asymétrique, de configuration (R,S), (R) ou (S),
- **C représente un atome de carbone asymétrique de configuration
(R,S) ou (R), selon un ou plusieurs traitements réactionnels connus de l'homme de l'art, pour obtenir le remplacement de tous les groupes R1, R2, R3, R' par un atome d'hydrogène.According to the invention, a process for the preparation of the compounds of formula (I) and of their addition salts, said process comprising the deprotection of a compound of formula, is also recommended.

in which:
R represents a hydrogen atom, an OCH 3 group, an OH group,
a CH 2 OH group, an OR 'group or a CH 2 OR group,
R 'represents a group protecting the hydroxyl function,
- R1, R2, R3, identical or different, each represent a group
protective of the amine function,
- * C represents, when R is not the hydrogen atom, an atom of
asymmetric carbon, of (R, S), (R) or (S) configuration,
- ** C represents an asymmetric carbon atom of configuration
(R, S) or (R), according to one or more reaction treatments known to those skilled in the art, to obtain the replacement of all the groups R1, R2, R3, R 'with a hydrogen atom.

 It is also recommended to use a substance chosen from the compounds of formula I and their non-toxic addition salts for obtaining medicinal products intended for therapeutic use for the treatment or the prevention of immune disorders or of malaria, or for use in the field of analysis as a pharmacological reagent.

Detailed description of the invention
By "addition salts" is meant here acid addition salts obtained by reaction of a compound of formula I with a mineral acid or an organic acid. The preferred mineral acids for salification are hydrochloric, hydrobromic, phosphoric and sulfuric acids. The preferred organic acids for salification are fumaric, maleic, methanesulfonic, oxalic, citric and trifluoroacetic acids.

As indicated in formula I, the compounds according to the invention comprise a carbon noted * C which, when R is not a hydrogen atom, is an asymmetric carbon, and a second carbon atom noted ** C which is an asymmetric carbon. When R represents a hydrogen atom, the present invention includes, among the compounds of formula
I, the racemic where ** C is of configuration (R, S) and the enantiomer where ** C is of configuration (R) .When R is not a hydrogen atom, the present invention encompasses, among the compounds of formula I which then have two centers of chirality, (1) the product of [(R, S) - * C (R, S) - ** C] configuration which is a substantially equimolecular mixture of the four stereoisomers, (2) ) "hemiracemic" products of configuration [(R, S) * C; (R) - ** C], [(R) - * C; (R, S) - ** C] and [(S) - * C; (R, S) - ** C], and (3) diastereoisomers of [(R) - * C configuration; (R) - ** C] and [(S) - * C; (R) - ** C].

 In a practical way, the compounds of formula I according to the invention in which ** C is of (R) configuration are preferred.

 The compounds of formula I can be obtained according to methods known per se, by application of conventional reaction mechanisms, in particular reactions commonly used in peptide chemistry, making it possible to obtain amide type bonds.

 The process for preparing the compounds of formula I which is recommended according to the invention comprises, as indicated above, the deprotection of a compound of formula II.

In a practical way, the protective groups R 1, R 2, R 3 that will be replaced by a hydrogen atom are amino-protecting groups of a type known in the field of peptide chemistry for temporarily blocking the "amine" functions. totally substituted. Some of the groups that are suitable for this purpose include:
a) oxycarbonyl groups, such as for example
alkoxycarbonyl and benzyloxycarbonyl groups:
Boc: t-butyloxycarbonyl (or 1,1-dimethylethoxycarbonyl)
Fmoc: 9-fluorenylmethyloxycarbonyl
Z: benzyloxycarbonyl
Z (p: 4-chlorobenzyloxycarbonyl, or
Z (p: 4-methoxybenzyloxycarbonyl
on the one hand, and
b) benzyl groups such as, for example, the group
phenylmethyl (Bn), on the other hand.

 Among these amino-protecting groups, the preferred groups are Boc, Fmoc and Bn groups.

 When in formula I, the substituent R comprises a hydroxyl function, it may be necessary to protect it to carry out the reactions leading to the compounds of formula II.

In this case, R can represent intermediately in the formula
II, a group OR 'or CH20R' wherein R 'is a hydroxyl protecting group.

Among these protecting groups for the hydroxyl function, mention may be made in particular of:
a) benzyl groups such as, for example, the group
phenylmethyl (Bn),
b) trialkylsilyl groups, such as groups
trimethylsilyl or (tert.-butyl) (dimethyl) silyl (tBDMS) from
formula:

and
c) the 2-tetrahydropyranyl group.

dans laquelle::
- R représente un atome d'hydrogène, un groupe O-CH3, un groupe OH,
un groupe CH2OH, un groupe OR' ou un groupe CH2OR',
- R' représente un groupe trialkylsilyle ou un groupe phénylméthyle,
- R1, R2, R3, identiques ou différents, représentent chacun un groupe
amino-protecteur de type alkoxycarbonyle ou benzyle,
- *C représente, lorsque R n'est pas l'atome d'hydrogène, un carbone
asymétnque de configuration (R,S), (R) ou (S),
- **C représente un carbone asymétrique de configuration (R,S) ou (R), selon un ou plusieurs traitements suivant la nature des groupes protecteurs, comme par exemple, si l'un au moins des groupes R1, R2, R3 et R' représente un groupe du type alcoxycarbonyle ou trialkylsilyle, par action d'un acide fort tel que notamment l'acide trifluoroacétique, ou, si l'un au moins des groupes R1, R2, R3 ou R' représente un groupe du type benzyle, par hydrogénation catalytique en présence d'un sel de palladium ou de charbon palladium, pour obtenir un composé de formule I sous forme de base libre ou de l'un de ses sels d'addition, et, si nécessaire (ii) à partir d'un sel d'addition obtenu selon l'étape (i), obtenir le composé de formule I sous forme de base libre par action d'une base forte, puis obtenir à partir de ladite base libre les autres sels d'addition.Conveniently, the process for preparing a compound of formula I or an addition salt thereof is characterized by comprising the steps of (i) deprotecting a compound of formula II:

in which::
R represents a hydrogen atom, an O-CH 3 group, an OH group,
a CH 2 OH group, an OR 'group or a CH 2 OR group,
R 'represents a trialkylsilyl group or a phenylmethyl group,
- R1, R2, R3, identical or different, each represent a group
amino-protecting agent of the alkoxycarbonyl or benzyl type,
- * C represents, when R is not the hydrogen atom, a carbon
asymmetrical configuration (R, S), (R) or (S),
- C represents an asymmetric carbon configuration (R, S) or (R), according to one or more treatments depending on the nature of the protecting groups, for example, if at least one of the groups R1, R2, R3 and R 'represents a group of the alkoxycarbonyl or trialkylsilyl type, by the action of a strong acid such as in particular trifluoroacetic acid, or, if at least one of the groups R1, R2, R3 or R' represents a benzyl type group; by catalytic hydrogenation in the presence of a palladium salt or palladium charcoal to obtain a compound of formula I in free base form or an addition salt thereof, and if necessary (ii) from of an addition salt obtained according to step (i), obtaining the compound of formula I in free base form by action of a strong base, then obtaining from said free base the other addition salts.

dans laquelle R1 représente un groupe amino-protecteur, comme par exemple un groupe Boc, avec un dérivé d'aminoacide de formule:

dans laquelle::
- R représente un atome d'hydrogène ou un groupe CH2OR',
- R' représente un groupe protecteur de la fonction hydroxyle, comme
par exemple un groupe tBDMS,
- R4 représente un groupe alkyle en C1-C3, et,
- *C, lorsque R n'est pas l'atome d'hydrogène, représente un atome de
carbone asymétrique de configuration (R,S) (racémique), (R) ou (S), par activation de la fonction acide à l'aide d'un agent de couplage de type carbodiimide, notamment le 1,3-dicyclohexylcarbodiimide (DCC), en présence d'un agent nucléophile, notamment le l-hydroxybenzotriazole (HOBT), dans un solvant organique comme par exemple du dichlorométhane, à une température comprise entre 0 et 40C C, et à raison de 1 mole du composé m pour environ 1 mole du composé IV, pour obtenir un composé de formule::

dans laquelle R, R1, R4 et *C conservent les mêmes significations que cidessus; (ii) hydrolyser la fonction ester du composé de formule V ainsi obtenu, soit selon l'étape (i) ci-dessus lorsque R représente l'atome d'hydrogène ou un groupe CH2OR' tel que décrit ci-dessus, soit selon un procédé connu lorsque R représente un groupe OR', où R' représente un groupe protecteur de la fonction hydroxyle comme par exemple le groupe tBDMS, par action d'une solution diluée d'une base, comme par exemple de l'hydroxyde de sodium, en présence d'un solvant miscible à l'eau comme par exemple du 1 ,2-diméthoxyéthane, à une température voisine de la température ambiante et pendant environ 2 à 30 minutes pour obtenir un composé de formule::

dans laquelle R1 et *C conservent la même signification que dans le composé de formule V, et R représente un atome d'hydrogène, un groupe CH2OR' ou un groupe OR', R' représentant un groupe protecteur de la fonction hydroxyle; (iii) faire réagir un composé de formule VI obtenu selon l'étape (ii) ci-dessus avec un composé de formule:

dans laquelle:
- R2 et R3, identiques ou différents, représentent chacun un groupe
amino-protecteur comme par exemple un groupe Boc ou un groupe Bn,
- **C représente un carbone asymétrique de configuration (R,S) ou (R), dans des conditions identiques à celles décrites à ltétape (i) ci-dessus pour obtenir un composé de formule Il:

dans laquelle R, R1, R2, R3, *C et **C conservent la même signification que ci-dessus ; et,
(b) la variante B qui comprend les étapes consistant à: (i) condenser un drivé d'aminoacide de formule

dans laquelle:
- R représente un atome d'hydrogène ou un groupe CH2OR' où R' est un
groupe protecteur de la fonction hydroxyle,
- R5 représente un groupe protecteur de la fonction amine, comme par
exemple un groupe Fmoc,
- *C représente, lorsque R n'est pas l'atome d'hydrogène, un atome de
carbone asymétrique de configuration (R,S), (R) ou (S),
avec un composé de formule: :

dans laquelle
- R2 et R3, identiques ou différents, représentent chacun un groupe
protecteur d'amine, tous deux différents du groupe protecteur R5
présent dans le composé de formule VIII, et
- **C représente un carbone asymétrique de configuration (R,S) ou (R), dans des conditions analogues à celles du procédé de ltétape (i) de la variante
A cidessus, pour obtenir un composé de formule::

dans laquelle R, R2, R3, Rg, *C et **C conservent la même signification que cidessus; (ii) déprotéger le composé IX ainsi obtenu, selon un procédé spécifique à la coupure de la liaison N-Rs comme par exemple, si R5 est un groupe Fmoc, un traitement avec de la pipéridine, dans un solvant, comme par exemple du dichlorométhane, à température ambiante et pendant environ 1 à 5 heures, pour obtenir un composé de formule:

dans laquelle R, R2 R3, *C et **C conservent la même signification que cidessus ; et, (iii) condenser l'amine de formule X ainsi obtenue avec un acide de formule:

dans laquelle R1 représente un groupe protecteur d'amine, comme par exemple un groupe Boc, dans des conditions opératoires analogues à celles décrites pour le procédé (i) de la variante A, pour obtenir un composé de formule fi:

dans laquelle R, R1, R2, R3, *C et **C conservent la même signification que cidessus. For the preparation of a compound of formula II, it is possible to use a process chosen from the following variants
(a) variant A which comprises the steps of: (i) condensing an acid of formula

wherein R1 represents an amino protecting group, such as for example a Boc group, with an amino acid derivative of formula:

in which::
R represents a hydrogen atom or a CH 2 OR group,
R 'represents a group protecting the hydroxyl function, as
for example a group tBDMS,
R4 represents a C1-C3 alkyl group, and,
- * C, when R is not the hydrogen atom, represents an atom of
asymmetric carbon configuration (R, S) (racemic), (R) or (S), by activation of the acid function using a carbodiimide coupling agent, especially 1,3-dicyclohexylcarbodiimide (DCC) ), in the presence of a nucleophilic agent, especially 1-hydroxybenzotriazole (HOBT), in an organic solvent such as dichloromethane, at a temperature of between 0 and 40 ° C., and at a concentration of 1 mol of the compound m for about 1 mole of compound IV, to obtain a compound of formula:

wherein R, R 1, R 4 and C retain the same meanings as above; (ii) hydrolyzing the ester function of the compound of formula V thus obtained, or according to step (i) above when R represents the hydrogen atom or a CH2OR 'group as described above, or according to a known method when R represents a group OR ', where R' represents a group protecting the hydroxyl function, for example the tBDMS group, by the action of a dilute solution of a base, such as, for example, sodium hydroxide, in the presence of a water-miscible solvent such as, for example, 1, 2-dimethoxyethane, at a temperature of about room temperature and for about 2 to 30 minutes to obtain a compound of formula:

wherein R1 and C retain the same meaning as in the compound of formula V, and R represents a hydrogen atom, a CH2OR 'group or an OR' group, R 'representing a hydroxyl protecting group; (iii) reacting a compound of formula VI obtained according to step (ii) above with a compound of formula:

in which:
- R2 and R3, identical or different, each represent a group
amino-protector such as for example a Boc group or a Bn group,
- ** C represents an asymmetric carbon of (R, S) or (R) configuration under conditions identical to those described in step (i) above to obtain a compound of formula II:

wherein R, R 1, R 2, R 3, C and C retain the same meaning as above; and,
(b) variant B which comprises the steps of: (i) condensing an amino acid derivative of formula

in which:
R represents a hydrogen atom or a CH2OR 'group where R' is a
protecting group of the hydroxyl function,
- R5 represents a protective group of the amine function, as per
example an Fmoc group,
- * C represents, when R is not the hydrogen atom, an atom of
asymmetric carbon configuration (R, S), (R) or (S),
with a compound of formula:

in which
- R2 and R3, identical or different, each represent a group
amine protector, both different from the R5 protecting group
present in the compound of formula VIII, and
- ** C represents an asymmetric carbon of (R, S) or (R) configuration under conditions analogous to those of step (i) of the variant
Above, to obtain a compound of formula:

wherein R, R2, R3, R8, C and C retain the same meaning as above; (ii) deprotecting the compound IX thus obtained, according to a specific method for cleaving the N-Rs bond, for example, if R5 is a Fmoc group, a treatment with piperidine, in a solvent, for example dichloromethane at room temperature and for about 1 to 5 hours, to obtain a compound of formula:

wherein R, R2, R3, C and C retain the same meaning as above; and (iii) condensing the amine of formula X thus obtained with an acid of formula:

in which R 1 represents an amine protecting group, such as for example a Boc group, under similar operating conditions to those described for process (i) of variant A, to obtain a compound of formula fi:

wherein R, R 1, R 2, R 3, C and C retain the same meaning as above.

 The acid of formula III wherein R 1 is 1,1-dimethylethoxycarbonyl (Boc) is prepared by reacting 7-aminoheptanol with N, N'-bis (Boc) -S-methylisothiourea and then oxidizing the alcohol. thus obtained by pyridinium dichromate in dimethylformamide.

dans laquelle
- Bn est le groupe phénylméthyle,
- Boc est le groupe 1, l-diméthyléthoxycarbonyle,
- **C est un atome de carbone asymétrique de configuration (R,S) ou
(R), par hydrogénation catalytique en présence de charbon palladium, ce qui permet de remplacer le groupe Bn par un atome d'hydrogène, puis action du dicarbonate de di(tert.-butyle) ce qui conduit au composé de formule:

et enfin, hydrogénation catalytique du groupe cyano, en présence de nickel de Raney, ce qui permet d'obtenir le composé de formule VII attendu.The compounds of formula VII in which R 2 and R 3 are identical and each represents the Boc group may be obtained starting from a compound of formula:

in which
Bn is phenylmethyl
Boc is the 1, 1-dimethylethoxycarbonyl group,
- ** C is an asymmetric carbon atom of configuration (R, S) or
(R), by catalytic hydrogenation in the presence of palladium-carbon, which makes it possible to replace the group Bn with a hydrogen atom, then the action of di (tert.-butyl) dicarbonate, which leads to the compound of formula:

and finally, catalytic hydrogenation of the cyano group, in the presence of Raney nickel, which makes it possible to obtain the compound of formula VII expected.

The compound of formula VII in which R2 and R3 are each
Boc is new and is one of the objects of the invention. It acts as a synthetic intermediate for obtaining the compounds of formula I according to the invention.

 The invention will be better understood on reading the examples which follow and the results of pharmacological tests obtained with the compounds according to the invention, compared with the results obtained with products known from the prior art. The nomenclature used in the examples is that recommended by Chemical Abstracts, so an ester of the type "t-butyl acetate" will be written in the form "acidic acid, 1,1-dimethylethyl ester" .

 In the experimental part, the preparations are relative to the intermediates and the examples relate to the products according to the invention.

 When the compounds comprise in their structure an asymmetric carbon, the absence of any particular indication or the indication (R, S) means that it is a substantially equimolecular mixture of the two enantiomers (ie "racemic" compound) . When these same compounds are denoted with a sign (R) or (S) immediately following the identification of the position of a substituent, it means that the carbon bearing this substituent is of (R) or (S) configuration. ), in accordance with the rules of Cahn, Ingold and Prelog.

 When the compounds have in their structure two centers of asymmetry, the absence of any particular indication or the presence of the sign (R, S) immediately following the positional identifications of the substituents means that it is a mixture of four stereoisomers. When these same compounds are denoted with a sign (R) or (S) immediately following the positional identification of a substituent, it means that the carrier carbon of this substituent is of determined configuration (R) or (S ): if only one of the centers of asymmetry is named with a sign of chirality, the product described will be a substantially equimolar mixture of two diastereoisomers. If the two centers of asymmetry are named with a sign of chirality, the described product is a pure stereoisomer.

 The spectral characteristics of the nuclear magnetic resonance (NMR) signals are given for the proton (1H) or for the carbon isotope 13 (13C): the chemical shift with respect to the tetramethylsilane signal is indicated and, in parentheses, the shape signal (s for singlet, d for doublet, t for triplet, q for quadruplet, m for multiplet, sl for wide signal) and the number of protons involved in the signal. As an indication, the 1 H NMR spectra were made at 300 MHz.

PREPARATION I
Acid (7-hydroxy-heptylcarbonimidoyl) bis (carbamic), bis (1,1-dimethylethyl) ester.

 4.57 g (35.10-3 mol) of 7-aminoheptanol and 10.15 g (35.10 -3 mol) of [[[(1,1-dimethylethoxy) -carbonyl] amino] (methylthio) methylene are dissolved. carbamic acid, 1,1-dimethylethyl ester in 400 ml of tetrahydrofuran and stirred for 15 hours at room temperature. The reaction mixture is concentrated under reduced pressure and purified by chromatography on silica gel, eluting with a methylcyclohexane / ethyl acetate mixture (7/3 v / v). 12.6 g of the expected product are thus obtained in the form of a yellow oil (yield = 96%).

1H NMR (CDCl3) 1.25-1.6 (m, 28H); 3.35-3.45 (q, 2H); 3.63 (q, 2H) 8.25 (t, 1H); 11.5 (s, 1H).

PREPARATION II
[(6-Carboxy-hexyl) carbonimidoyl] bis (carbamic) acid, bis (1,1-dimethylethyl) ester.

 12.6 g (33.7 × 10 -3 mol) of the compound obtained according to Preparation I are dissolved in 100 ml of dimethylformamide and 25.2 g (67.103 mol) of pyridinium dichromate are added. The reaction medium is stirred for 2 hours at room temperature and then hydrolyzed on 1 l of water. It is extracted 3 times with diethyl ether and the combined organic phases are washed with a solution of copper sulphate and then with water.

The organic phase is then dried over magnesium sulfate and then concentrated under reduced pressure. 12 g of the expected product are thus obtained in the form of an oil (yield = 91.6%).

1H NMR (CDCl3): 1.25-1.70 (m, 26H); 2.35 (t, 2H); 3.40 (q, 2H), 8.30 (t, 1H); 11.5 (sl, 1H).

PREPARATION m
3 - [[(1,1-dimethylethoxy) carbonyl] amino] -1H-oxo-2,4,12-triazetetradec-2-enedioic acid, 1- (1,1-dimethylethyl) ethyl ester.

 A solution of 5.28 g (13.64 × 10 -3 mol) of the acid obtained according to Preparation II in 80 ml of dichloromethane is prepared. The solution is cooled to 0 ° C. and 1.84 g (13.64 × 10 -3 mol) of 1-hydroxybenzotriazole hydrate (HOBT), 5.63 g (27.28 × 10 -3 mol) of N, N'-dicyclohexylcarbodiimide ( DCC) and then a solution of 2.1 g (15 × 10 -3 moles) of ethyl glycinate hydrochloride and 1.51 g (15 × 10 -3 moles) of triethylamine in 20 ml of dichloromethane. The reaction mixture is stirred for 48 hours at room temperature and then concentrated under reduced pressure. The residue is then purified by chromatography on silica gel, eluting with a methylcyclohexane / ethyl acetate mixture (6/4) to give 4.41 g of the expected product in the form of a colorless oil (Yield = 68.6%). %).

1H NMR (CDCl3): 1.25 (t, 3H); 1.3-1.8 (m, 26H); 2.25 (t, 2H) 3.4 (t, 2H); 4.0 (d, 2H); 4.2 (q, 2H); 6.0 (sl, 1H); 8.3 (t, 1H); 11.5 (s, 1H).

PREPARATION IV
3 - [[(1,1-dimethylethoxy) carbonyl] amino] -11-oxo-2,4,12-triazetetetradec-2-enedioic acid, 1- (1,1-dimethylethyl) ester.

 4.41 g (9.34 × 10 -3 mol) of the compound obtained according to Preparation III are dissolved in 15 ml of 1,2-dimethoxyethane, and 15 ml of 1N sodium hydroxide are then added. Stirred at room temperature (20-25 ° C.) for 30 minutes, then 100 ml of dichloromethane are added and the mixture is slowly acidified, with cooling and with good stirring, with 1N hydrochloric acid, to pH = 1. Decanting the organic phase and then the aqueous phase is extracted twice with 100 ml of dichloromethane. The combined organic phases are dried over magnesium sulphate and then concentrated under reduced pressure. 4.1 g of the expected product are thus obtained in the form of a yellow oil (yield = 99%).

1H NMR (CDCl3): 1.3-1.8 (m, 26H); 2.25 (t, 2H); 3.3-3.35 (m, 2H) 4.0 (d, 2H); 6.5 (t, 1H); 8.45 (sl, 1H); 11.5 (s, 1H).

PREPARATION V
3-L (1,1-dimethylethoxy) carhonyllamino] -23 (R) -methyl-20-phenylmethyl-11,14-dioxo-2,4,12,15,20,24-hexaazapentacos-2-enedioic acid bis (1,1-dimethylethyl) ester.

4.1 g (9.23 × 10 -3 mol) of acid obtained according to Preparation IV are dissolved in 100 ml of dichloromethane, then 1.35 g (10 × 10 -3 mol) of HOBT and 4.13 g are added ( 20.10-3 mol) of DCC and stirred at 0.degree.
C for 30 minutes. 3.3 g (9.45 × 10 -3 moles) of [3 - [(4-aminobutyl) (phenylmethyl) amino] -1 (R) -methylpropyl] carbamic acid, 1,1-dimethylethyl ester are then added, followed by stirring for 24 hours at room temperature. After concentration of the reaction medium under reduced pressure, the residue is purified by chromatography on silica gel in Quant with an ethyl acetate / ethanol (9/1) mixture. 5.9 g of the expected product are thus obtained in the form of an amorphous white solid (yield = 82.5%).

[?] D24.5 = + 0.44 (c = 0.45, CHCl3)
1H NMR (DMSO d6): 0.95 (d, 3H); 1.2-1.7 (m, 41H); 2.1 (t, 2H) 2.3-2.45 (m, 4H); 3.0-3.1 (m, 2H); 3.2-3.4 (m, 4H); 3.4-3.5 (m, 1H) 3.6 (d, 2H); 6.65 (d, 1H); 7.25-7.35 (m, SH); 7.7 (t, 1H); 7.95 (t, 1H); 8.3 (t, 1H); 11.5 (s, 1H).

PREPARATION VI
3 - [[(1,1- (dimethylethoxy) carbonyl] amino] -23 (R) -methyl-11,14-dioxo-2, 4, 12, 15,20,2hexaazapentacos-2-enedioic acid, bis (I) 1-dimethylethyl) ester.

 5.9 g (7.61 × 10 -3 mol) of the compound obtained according to Preparation V are dissolved in 120 ml of pure ethanol, 500 mg of 5% palladium-on-carbon are added and the mixture is stirred under an atmosphere of hydrogen for 8 hours. hours, at room temperature. The catalyst is then removed by filtration and the filtrate is concentrated under reduced pressure. 4.94 g of the expected product are thus obtained in the form of an amorphous solid (yield = 95%).

[a] D24 = - 4.8 (c = 1.00, CHCl3)
1H NMR (DMSO d6): 1.05 (d, 3H); 1.2-1.8 (m, 41H); 2.17 (t, 2H) 2.7-2.85 (m, 4H); 3.0-3.15 (m, 2H); 3.25-3.4 (m, 3H); 3.45-3.55 (m, 1H) 3; 6 (d, 2H); 6.35 (d, 1H); 7.85 (t, 1H); 8.0 (t, 1H); 8.3 (t, 1H); 11.5 (sl, 1H).

Example 1 N- [2 [[4 - [(3 (R) -aminobutyl) amino] butyl] amino] -2-oxoethyl] -7 - [(aminoiminomethyl) amino] heptanamide, tris (trifluoroacetate).

 A mixture of 4 g (5.83 × 10 -3 mol) of the compound obtained according to Preparation VI in 25 ml of dichloromethane and 25 ml of trifluoroacetic acid is prepared and stirred at room temperature for 5 hours. The reaction mixture is then concentrated under reduced pressure and the evaporation residue is purified by medium pressure chromatography (MPLC), using a grafted silica gel of RP18 type with a particle size of 5 to 20 μm. The eluent is a mixture of acetonitrile, water and trifluoroacetic acid in the proportions 0.8 / 8 / 1.2 by volume. The pure fractions are lyophilized and the solid obtained is redissolved in 100 ml of water. The solution obtained is washed three times with ethyl acetate and then freeze-dried. This gives 3.0 g of the expected product in the form of a white amorphous solid (yield = 70%).

[a] D23 = + 1.3 (c = 1.00, CH30H)
1H NMR (DMSO d6) 1.18 (d, 3H); 1.2-1.6 (m, 12H); 1.65-1.85 (m, 1H) 1.85-2.0 (m, 1H); 2.10 (t, 2H); 2.85-3.1 (m, 8H); 3.2-3.4 (m, 1H) 3.65 (d, 2H); 6.9-8.5 (m, 12H).

13 C NMR (D 2 O / Dioxane Hg): 18.02; 23.68; 25.81; 26.31; 26.37 28.51; 28.61; 31.23; 36.19; 39.22; 41.89; 43.44; 44.61; 46.10; 48.16; 158.20; 172.37; 178.71.

Example 2
N- [2 [[4 - [(3 (R, S) -aminobutyl) amino] butyl] amino] -2-oxoethyl] -7 - [(aminoiminomethyl) amino] heptanamide, tris (trifluoroacetate).

 By following a procedure analogous to the synthesis scheme used for Example 1, but using [3 - [(4-aminobuty) -phenylmethyl) aminol-1 (R, S) -methylpropyl] carbamic acid, 1 1-dimethylethyl ester, the expected product is obtained in the form of an amorphous white solid.

1H NMR (DMSO d6) 1.18 (d, 3H); 1.2-1.35 (m, 4H); 1.4-1.60 (m, 8H) 1.65-1.85 (m, 1H); 1.85-2.0 (m, 1H); 2.10 (t, 2H); 2.8-3.2 (m, 8H) 3.2-3.4 (m, 1H); 3.65 (d, 2H); 6.8-8.7 (m, 12H).

13 C NMR (D 2 O + Dioxane Hg) 18.01; 23.68; 25.81; 26.31; 26.37 28.51; 28.60; 31.23; 36.18; 39.21; 41.88; 43.44; 44.61; 46.03 48.16; 157.54; 172.38; 178.72.

PREPARATION VII
[3 - [(3-cyanopropyl) amino] -1 (R) -methylpropyl] carbamic acid, 1,1-dimethylethyl ester.

21 g (60.8 × 10 -3 moles) of [3 - [(3-cyanopropyl) - (phenylmethyl) amino] -1 (R) -methylpropyl] carbamic acid, 1,1-dimethylethyl ester are dissolved in 400 ml of ethyl acetate. ethanol and then 0.25 ml of hydrochloric acid is added
M, then 1.2 g of 5% palladium on carbon. The mixture is stirred under an atmosphere of hydrogen at ambient temperature, at atmospheric pressure.

After 24 hours of reaction, the catalyst is removed by filtration and the filtrate is concentrated under reduced pressure. The residue is purified by chromatography on silica gel, eluting with a mixture of ethyl acetate / ethanol / ammonia (6/3 / 0.1 v / v). 6 g of the expected product are thus obtained in the form of an oil (yield = 34%).

1H NMR (CDCl3): 1.15 (d, 3H); 1.3-1.9 (m, 13H); 2.4-2.7 (m, 6H) 3.4-3.85 (m, 1H); 4.7-4.9 (sl, 1H).

PREPARATION VIII
[3 - [(3-cyanopropyl) [(1,1-dimethylethoxy) carbonyl] amino] -1 (R) -methylpropyl] carbamic acid, 1,1-dimethylethyl ester.

A solution of 5.82 g (22.92 × 10 -3 mol) of the compound obtained according to Preparation VII in 100 ml of tetrahydrofuran is prepared and 3.45 g (34.23 × 10 -3 mol) of triethylamine and then 5.97 are added. g (27.38.1t3 mol) of di (tert.-butyl) dicarbonate (chemical structure
O [CO2C (CH3] 2). The reaction mixture is stirred for 15 hours at room temperature. After concentration of the reaction medium under reduced pressure, the crude product is purified by chromatography on silica gel, eluting with a methylcyclohexane / ethyl acetate mixture (9/1 and then 8/2 v / v). 5.6 g of the expected product are thus obtained in the form of an oil (yield = 68%).

1H NMR (CDCl3): 1.15 (d, 3H); 1.4-1.6 (m, 20H); 1.8-1.95 (m, 2H) 2.35 (t, 2H); 3.1-3.4 (m, 4H); 3.55-3.70 (m, 1H); 4,254.55 (sl, 1H).

PREPARATION Ix
[3 - [(eaminobutyl) [(1,1-dimethylethoxy) carbonyl] -1- (R) -methylpropyl] carbamic acid, 1,1-dimethylethyl ester.

5.5 g (15.5 × 10 -3 moles) of the compound obtained according to Preparation VIII are dissolved in 100 ml of methanol, 400 mg of nickel are added.
Raney and the mixture is stirred under a hydrogen atmosphere at room temperature under a pressure of 3.105 Pascals for 24 hours. The reaction mixture is then filtered to remove the catalyst and then concentrated under reduced pressure. The residue is purified by chromatography on silica gel by netting with a methylcyclohexane / ethyl acetate mixture (6/4 v / v) and then ethyl acetate / ethanol / ammonia (6/3 / 0.2 v / v) . 3.96 g of the expected product are thus obtained in the form of an oil (yield = 71%).

[a] D21 = + 41.3 (c = 3; CHCl3)
1H NMR (CDCl3): 1.15 (d, 3H); 1.4-1.7 (m, 22H); 1.8 (s, 4H); 2.75 (t, 2H); 3.05-3.35 (m, 4H); 3.55-3.7 (m, 1H); 4,354.6 (sl, 1H).

PREPARATION X
3 - [[(1,1-dimethylethoxy) carbonylamino] -13- (carboxy) -15,15,16,16-tetramethyl-11-oxo-14-oxa-2,4,12-triaza-15-silaheptadec acid -2-ennoic acid, 1,1-dimethylethyl ester.

 1.73 g (2.94 × 10 -3 moles) of 3 - [[1,1-dimethylethoxy] carbonyl] amino] -13- (methoxycarbonyl) -15,15,16,16-tetramethyl- 11-oxo-2,4,12-triaza- 15-silaheptadec-2-ene, 1,1-dimethylethyl ester in 4 ml of 1,2-dimethoxyethane and 4 ml of a molar aqueous solution of sodium. The reaction mixture is stirred for 10 minutes at room temperature, then 20 ml of water and 20 ml of dichloromethane are added and the mixture is acidified with good stirring to pH 2 using 1N hydrochloric acid. After separation of the organic phase, the aqueous phase is extracted with 3 times 25 ml of dichloromethane. The combined organic phases are dried over magnesium sulphate and then concentrated under reduced pressure. In this way 1.68 g of the expected product is obtained in the form of a colorless oil (quantitative yield).

1H NMR (CDCl3): 0.1-0.15 (m, 6H); 0.85-1 (m, 9H); 1.3-1.8 (m, 26H) 2.25 (t, 2H); 3.2-3.45 (m, 2H); 5.55 (d, H); 7.0-7.1 (sl, 1H) 8.3-8.7 (m, 1H); 11-12 (sl, 1H).

 PREPARATION XI 3 - [[(1,1-Dimethylethoxy) carbonyl] amino] -20 - [(1,1-dimethylethoxy) carbonyl] -23 (R) -methyl-13 - [[(1,1-dimethylethyl) ) dimethylsilyl] oxy] -11,14-dioxo-2,4,12,15,2O, 24-hexaazapentaoes-2edioic acid, bis (1,1-dimethylethyl) ester.

 1.68 g (2.92 × 10 -3 mol) of the acid obtained according to Preparation X are dissolved in 30 ml of dichloromethane and cooled to 0 ° C. 0.4 g (3.10 -3 mol) of HOBT are added, then 1.24 g (6.10-3 mol) of DCC. This mixture is stirred for 15 minutes, then 1.05 g (2.92 × 10 -3 mol) of the compound obtained according to Preparation IX are added and the reaction medium is stirred at room temperature for 48 hours. The solvent is then removed under reduced pressure and the residue is purified by chromatography on silica gel, eluting with ethyl acetate and methylcyclohexane (1/1 v / v) and then with ethyl acetate. 0.78 g of the expected product is thus obtained in the form of an amorphous solid (yield = 30%).

[a] D19.5 = - 0.1 (c = 1; CHCl3)
1H NMR (DMSO d6): 0.079 (s, 3H); 0.137 (s, 3H); 0.88 (s, 9H) 1; 05 (d, 3H); 1.1-1.95 (m, 50H); 2.14 (t, 2H); 3-3.4 (m, 9H); 5.60 (d, 1H) 6.7 (sl, 1H); 7.75-7.85 (sl, 1H); 8.3 (t, 1H); 8.62 (d, 1H); 11.5 (s, 1H).

Example 3
N- [2 - [(4 - [(3 (R) -aminohutyl) amino] butyl] amino] -1 (R, S) -methoxy-2-oxoethyl] 4-7 - [(aminoiminomethyl) amino] heptanamide, tris (trifluoroacetate).

31 mg (38.7 × 10 -6 mol) of the compound obtained are dissolved in the mixture
XI in 10 ml of trifluoroacetic acid and 1 ml of methanol. This mixture is then kept stirring for 4 hours and then the solvents are removed under reduced pressure at room temperature. The crude product is then purified by chromatography on grafted silica of RP 18 type (particle size: 5 to 20 m), eluting with an acetonitrile / water / trifluoroacetic acid mixture (1.5 / 8 / 0.5 then 2/8 / 0.1 v / v). 16 mg of the expected product are thus obtained in the form of an amorphous white solid (Yield = 55%).

[a] D20 = + 1.2 (c = 1.53; H 2 O)
1H NMR (DMSO d6) 1.18 (d, 3H); 1.2-1.35 (m, 4H); 1.35-1.65 (m, 8H) 1.65-1.85 (m, 1H); 1.85-2.0 (m, 1H); 2.1-2.3 (m, 2H); 2.8-3.2 (m, 8H) 3.22 (m, 4H); 5.26 (d, 1H); 6.8-7.4 (s, 3H); 7.55 (t, 1H); 7.8-8.05 (m, 4H); 8.15 (t, 1H); 8.48 (d, 1H); 8.5-8.65 (m, 2H).

Example 4
N- [2 - [[4 - [(3 (R) -aminobutyl) amino] butyl] amino] -1 (R, S) -hydroxy-2-oxoethyl] -7 - [(aminoiminomethyl) amino] heptanamide tris (trifluoroacetate).

150 mg (0.164 × 10 -3 mol) of the compound obtained according to Preparation XI are dissolved in 4 ml of trifluoroacetic acid and stirred at room temperature for 4 hours. After removal of the solvent under reduced pressure, the product is purified by FPLC (Fast Protein Liquid Chromatography) technique on a CM Sepharose-type gel.
Fast Flow (Pharmacia) eluting with pure water and then with a solution of sodium chloride, the concentration of which increases gradually from 0 to 1 M according to a substantially linear gradient, with a concentration step at 0.4 M. Fractions containing the expected product are lyophilized and the white solid obtained is desalted by chromatography on a column of SEPHADEX # LH 20 (Pharmacia) eluting with methanol. The final purification of the product is carried out by silica gel chromatography graft type RP 18 "Varian Bond Elut" eluting with pure water, then with a mixture acetonitrile / water / trifluoroacetic acid (7 / 0.5 / 0.2 v / v). 10 mg of the expected product are thus obtained in the form of an amorphous white solid.

[a] D22 = + 1.5 (c = 0.45, CH3OH)
1 H NMR (D 2 O): 1.25-1.4 (m, 7H); 1.45-1.8 (m, 8H); 1.85-2.05 (m, 1H) 2.1-2.20 (m, 1H); 2.25 (t, 2H); 3.0-3.3 (m, 8H); 3.4-3.55 (m, 1H) 5.41 (s, 1H).

13 C NMR (H2O + Dioxane H8): 18.34; 23.70; 25.6; 26.12; 26.22 28.38; 28.51; 31.14; 36.33; 39.37; 41.94; 44.66; 46.28; 48.22 72.54; 157.25; 171.79; 178.41.

PREPARATION XII
3 - [[(1,1-dimethylethoxy) carbonyl] amino] -23 (R, S) -methyl-20 (phenylmethyl) -13 - [[(1,1-dimethylethyl) dimethylsilyloxy] -11,14-dioxo 2,4, 12, 15,20,24-hexaazapentacos-2inedioic acid, bis (1,1-dimethylethyl) ester.

 By proceeding analogously to the process of Preparation XI, but using [3 - [(4-aminobutyl) (phenylmethyl) amino] -1 (R, S) -methylpropyl] -carbamic acid, 1,1- dimethylethyl ester, the expected product is obtained in the form of an oil, with a yield of 49%.

1H NMR (CDCl3): 0.105 (s, 3H); 0.21 (s, 3H); 0.90 (s, 9H); 1.04 (d, 3H) 1.3-1.7 (m, 41H); 2.20 (m, 2H); 2.41 (m, 2H); 2.55 (m, 1H) 3.1-3.75 (m, 7H); 5.3 (sl, 1H); 5.7 (d, 1H) 6.4 (ss, 1H); 6.7 (sl, 1H) 7.20-7.30 (m, 5H); 8.3 (sl, 1H); 11.5 (s, 1H).

PREPARATION XIII
3 - [[(1,1-Dimethylethoxy) carbonyl] amino] -23 (R, S) -methyl-13 (R, S) - [[(1,1-dimethylethyl) dimethylsilyl] oxy] -11-14 2,4,12,15-dioxo-20,24-hexaazapentacos-2-enedioic acid, bis (1,1-dimethylethyl) ester.

 300 mg (0.33 × 10 -3 mol) of the compound obtained according to Preparation XII are dissolved in 30 ml of methanol and 30 mg of palladium hydroxide are added. The mixture is stirred under an atmosphere of hydrogen at atmospheric pressure and at room temperature for 15 min and then the catalyst is removed by filtration. After concentration, the crude product is purified by chromatography on silica gel, eluting with a mixture of ethyl acetate / ethanol / ammonia (6/3 / 0.1 v / v). In this way 200 mg of the expected product is obtained in the form of an oil (yield: 74%).

1H NMR (CDCl3): 0.011 (s, 3H); 0.021 (s, 3H); 0.9 (s, 9H); 1.13 (d, 3H) 1.3-1.9 (m, 41H); 2.20 (m, 2H); 2.7 (m, 4H); 2.7 (m, 4H); 3.1-3.6 (m, 5H); 4.24 (sl, 1H); 4.8 (sl, 1H); 5.7 (d, 1H); 6.7-6.9 (m, 2H) 8.3 (sl, 1H); 11.5 (s, 1H).

Example 5
N- [2 - [[4 - [(3 (R, S) -aminobutyl) amino] butyl] amino] -1 (R, s) -hydroxy-2-oxoethyl] -7 - [(aminoiminomethyl) amino] ] heptanamide, tris (trifluoroacetate).

 200 mg (0.245 × 10 -3 mole) of the compound obtained according to Preparation XIII are stirred in 10 ml of trifluoroacetic acetate at room temperature for 45 minutes. After evaporation of the solvent, the compound is purified by chromatography on grafted silica gel of RP 18 5-20N type, eluting with an acetonitrile / water / trifluoroacetic acid (2/8 / 0.1 v / v) mixture. After lyophilization, 82 mg of the expected product are obtained in the form of a white solid (yield: 45%).

1H NMR (DMSO): 1.15 (d, 3H); 1.2-1.35 (m, 4H); 1.35-1.55 (m, 8H) 1.65-1.8 (m, 1H); 1.85-2.0 (m, 1H); 2.1-2.2 (t, 2H); 2.85-3.35 (m, 9H) 5.4 (d, 1H); 6.5 (sl, 1H) 6.7-7.4 (nd, 3H); 7.6 (t, 1H) 7.8-8.1 (m, 5H) 8.4-8.7 (m, 3H).

PREPARATION xlv
131S) - [(9H-Fluoro-9-yl) -methoxycarbonyl] amino] -3 (R) -methyl-L2-oxo-14- (phenylmethoxy) -6- (phenylmethyl) -2,6,11 acid - triazatetradecanoic acid, 1,1-dimethylethyl ester.

 N-[(9H-fluoren-9-yl) methoxycarbonyl] -O-phenylmethyl- (L) -serine (1.71 g, 4.1 × 10 -3 mol) was dissolved in 60 ml of dichloro-methane. It is cooled to 0 ° C. and 0.55 g (4.10-3 mol) of HOBT and 1.54 g (7.5 × 10 -3 mol) of DCC dissolved in 20 ml of dichloromethane are added.

The mixture is stirred for 0.5 hours and then 1.30 g (3.72 × 10 -3 mol) of N- [3 - [(4-aminobutyl) (phenylmethyl) amino] -1- (R) are added. Methylpropylcarbamic acid, 1,1-dimethylethyl ester and the reaction mixture was allowed to stir at room temperature for 16 hours. The solvent is removed under reduced pressure and the residue is purified by chromatography on silica gel, eluting with a mixture of ethyl acetate / methylcyclohexane (1/1 v / v) and then with pure ethyl acetate. 2.7 g of the expected product are thus obtained in the form of a white crystalline solid (yield = 96%).

[?] D23 = +11 (c = 1.1, CHCl3) mp = 118 ° C
1H NMR (CDCl3): 1.02 (d, 3H); 1.3-1.8 (m, 15H); 2.25-2.6 (m, 4H) 3.15-3.3 (m, 2H); 3.35-3.75 (m, 4H); 3.8-3.95 (m, 1H); 4.154.6 (m, 6H) 5.3 (d, 1H); 5.6-5.8 (sl, 1H); 6.35-6.55 (sl, 1H); 7.15-7.45 (m, 14H) 7.55 (d, 2H); 7.75 (d, 2H).

PREPARATION XV
13 (S) -amino-3 (R) -methyl-12-oxo-14-phenylmethoxy-6-phenylmethyl-2,6,11-triazatetradecanoic acid, 1,1-dimethylethyl ester.

 2.56 g (3.42 × 10 -3 mol) of the compound obtained according to Preparation XIV are dissolved in 100 ml of dichloromethane and 5 g of piperidine are added. Stirring is maintained for 3 hours at room temperature and then concentrated under reduced pressure. 2 ml of toluene are added at the end of the concentration in order to expel the excess of piperidine. The residue is then purified by chromatography on silica gel, eluting with a mixture of ethyl acetate / methanol (8/2 v / v). 1.68 g of the expected product are thus obtained in the form of a yellow viscous oil (yield = 90%).

[a] D22 = + 10 (c = 1.1, CHCl3)
1H NMR (CDCl3) 1.03 (d, 3H); 1.4-1.7 (m, 15H); 2.3-2.65 (m, 4H) 3.15-3.30 (m, 2H); 3.4-3.8 (m, 6H); 4.5 (s, 2H); 5.35-5.55 (sl, 1H) 7.2-7.5 (m, 10H).

(Amine protons and amide undetected).

PREPARATION XVI
3 - [[(1,1-dimethylethoxy) carbonyl] anmino] -13 (S) - [phenylmethoxymethyl] -23 (R) -methyl-20-phenylmethyl-11,14-dioxo-2,4,12 acid , 15,20,24-hexaazapentacos-2-enedioic acid, bis (1,1-dimethylethyl) ester.

 1.46 g (3.79 × 10 -3 mol) of the acid obtained in Preparation II and 60 ml of dichloromethane are mixed and cooled to 0 ° C. 0.51 g (3.79 × 10 -3 mol) of HOBT and 1.56 g (7.56 × 10 -3 moles) of DCC, dissolved in 20 ml of dichloromethane. After stirring for 30 minutes, 1.60 g (3.10-3 mol) of the compound obtained according to Preparation XV are added and the mixture is stirred for 16 hours at room temperature. The reaction tit is then concentrated under reduced pressure and purified by chromatography on silica gel, eluting with a mixture of ethyl acetate / methylcyclohexane (6/4 v / v) and then pure ethyl acetate. 2.27 g of the expected product are thus obtained in the form of an oil (yield = 83%).

[a] D22 = + 4.2 (c = 4.7; CHCl3)
1H NMR (CDCl3) 1.03 (d, 3H); 1.2-1.8 (m, 41H); 2.2 (t, 2H); 2.3-2.65 (m, 4H); 3.1-3.8 (m, 8H); 3.85 (dd, 1H); 4.55 (q, 3H); 5.3-5.4 (m, 1H) 6.3-6.6 (m, 2H); 7.2-7.4 (m, 10H); ; 8.3 (t, 1H); 11.5 (s, 1H).

PREPARATION XVII
3 - [[(1,1-dimethylethoxy) carhonyl] amino] -13 (S) - [phenylmethoxymethyl] -23 (R) -methyl-11,14-dioxo-2,4,12,15,20 , 24-hexaazapentacos-2-enedioic acid, bis (1,1-dimethylethyl) ester.

 A solution of 1.65 g (1.84 × 10 -3 mol) of the compound obtained according to Preparation XVI is prepared in 100 ml of ethanol. 200 mg of 10% palladium on carbon are added and this reaction mixture is stirred under an atmosphere of hydrogen at atmospheric pressure for 24 hours at room temperature. The catalyst is removed by filtration and the filtrate is concentrated under reduced pressure. The residue is purified by chromatography on silica gel, eluting with a mixture of ethyl acetate / ethanol / ammonia (6/3 / 0.1 v / v). 1 g of the expected product is thus obtained in the form of yellow viscous oil (yield = 67%).

1H NMR (CDCl3) 1.15 (d, 3H); 1.3-1.9 (m, 41H); 2.2 (t, 2H); 2.5-2.7 (m, 4H); 3.2-3.3 (m, 2H); 3.4 (td, 2H); 3.5 (t, 1H); 3.65-3.75 (t, 1H); 3.85 (m, 1H); 4,454.65 (m, 3H); 4.84.9 (sl, 1H); 6.46.5 (sl, 1H); 6.45 (t, 1H); 7.25-7.4 (m, 5H); 8.3 (t, 1H); 11.5 (s, 1H).

PREPARATION XVIII
3 - [[(1, dimethylethoxy) carbonyl] amino] -13 (S) - (hydroxymethyl) - 23 (R) -methyl-1,1,4-dioxo-2,4,12,15,20,24- hexaazapentacos-2-eneediol that bis (1,1-dimethylethyl) ester.

 By following a procedure analogous to Preparation XVII, starting from the compound obtained according to Preparation XVII1 but carrying out the hydrogenation on an acidified reaction medium to pH 3.5 with concentrated hydrochloric acid, the expected product is obtained. with a yield of 95%.

[a] D22 = - 5.6 (c = 1.0, CHCl3)
1H NMR (CDCl3) 1.2 (d, 3H); 1.3-2.0 (m, 41H); 2.35 (t, 2H); 2.8-3.55 (m, 9H); 3.7-3.9 (m, 2H); 4.04.1 (m, 1H); 4.85-4.95 (s1.1H); 7.35 (t, 1H); 7.4 (d, 1H); 8.3 (t, 1H); 8.6-8.8 (sl, 1H); 9.3-9.5 (sl, 1H); 11.5 (s, 1H).

Example 6
N [2 - [[4 - [(3 (R) -aminobutyl] amino] butyl] amino] -1 (S) -hydroxymethyl-2-oxoethyl-7 - [(aminoiminomethyl) amino] heptanamide, tris (trifluoroacetate) .

 A solution of 1.10 g (1.54 × 10 -3 mol) of the compound obtained according to Preparation XVIII in 15 ml of dichloromethane is prepared, 15 ml of trifluoroacetic acid are added and the mixture is stirred for 16 hours at room temperature. The reaction medium is then concentrated under reduced pressure and purified by RP 18 silica gel chromatography, eluting with a water / trifluoroacetic acid / acetonitrile mixture (8/1/1 v / v). The fractions containing the pure product are freeze-dried and then taken up with 100 ml of distilled water, extracted with ethyl acetate and again lyophilized. 610 mg of the expected product is thus obtained in the form of a white amorphous solid (yield = 52%).

[a] D22 = - 3.2 (c = 1.0, CH30H)
1H NMR (DMSO d6): 1.2 (d, 3H); 1.2-1.35 (m, 4H); 1.4-1.6 (m, 8H) 1.70-1.85 (m, 1H); 1.85-2.0 (m, 1H); 2.14 (t, 2H); 2.85-3.15 (m, 8H) 3.25-3.4 (m, 1H); 3.55 (d, 2H); 4.2 (q, 1H); 4.8-4.95 (sl, 1H) 6.85-7.4 (sl, 3H); 7.65 (t, 1H), 7.80 (d, 1H); 7.9 (t, 1H); 7.95-8.1 (m, 3H) 8.5-8.8 (s, 3H).

13 C NMR (D2O + Dioxane H8): 17.77; 23.43; 25.58; 26.06; 26.09; 26.09 28.25; 28.35; 30.97; 35.86; 39.06; 41.65; 44.35; 45.81; 47.90 to 56.54; 61.65; 158.00; 172.54; 178.1.

 The immunosuppressive activity of the products according to the invention was demonstrated using a so-called graft versus host test.

Male B6D2F1 mice (first generation hybrids C57B1 / 6x
DBA / 2) are immunosuppressed by intraperitoneal (ip) injection of cyclophosphamide. Three days later (day 0 of the experiment: JO), they receive intravenously 4 x 107 splenocytes of C57B1 / 6 mice. The animals are then distributed in batches of at least 8 and receive a daily treatment of J1 to J5 and J7 to J10 by ip The control group receives the vehicle alone. Mortality is monitored until D60. The results, expressed as the mean value of survival in days at the indicated dose, are grouped in Table I where the values given are significant according to the Logrank test (probability less than or equal to 5%).

For the purpose of comparison, the values obtained with the products known from the prior art are also indicated in Table I: 15-deoxyspergualine in the form of tris (trifluoroacetate) (Product A) and the following known compounds
Product B:

3 CF3CO2H Prodiit C:

 3 CF3CO2H where * C is configuration S.

 This comparison shows that the products according to the invention are more active than the products known from the prior art.

 The products according to the invention are useful in therapy as curative or preventive immunosuppressive agents, in particular in the prevention of the rejection of allogeneic or xenogeneic vascular or non-vascular organs, of the graft-versus-host reaction following a vascularized graft. or not, in the treatment of auto-immune diseases genetically defined or acquired (such as for example systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis), chronic inflammatory diseases such as rheumatoid arthritis as well as in all pathologies where an immune disorder appears to be the cause or the factor responsible for maintaining a degraded clinical condition.

 The products according to the invention may also be administered in addition to cytotoxic anticancer drugs in order to limit the side effects and in addition to the administration of products derived from biotechnologies, in particular recombinant cytokines, mono- and polyclonal antibodies in order to decrease the appearance of protective antibodies produced by the patient.

 The products according to the invention can be used in the curative treatment of parasitoses, in particular in the case of malaria.

 The products according to the invention can be administered orally, by injection (especially intramuscularly or intravenously), topically (especially in the form of cream for topical application, eye drops), transdermally, rectally. in the form of suppository, or by inhalation.

 The products according to the invention are also useful as pharmacological reagents, particularly in the study of autoimmune diseases.

 In Table I, all the products cited examples of the invention are in the form of tris (trifluoroacetate).

3 CF3C02H

TABLE I

3 CF3C02H

<tb> Example <SEP> R <SEP> * C <SEP> ** C <SEP><SEP> Dose <SEP> Activity
<tb><SEP> (mg / Kg)
<tb><SEP> 1 <SEP> H <SEP> R
<tb><SEP> 2 <SEP> H <SEP> R, S <SEP> l <SEP><SEP> 42
<tb><SEP> 3 <SEP> OCH3 <SEP> R, S <SEP> R
<tb><SEP> 4 <SEP> OH <SEP> R, S <SEP> R
<tb><SEP> 5 <SEP> OH <SEP> R, S <SEP> R, S <SEP> 0.3 <SEP> 56
<tb><SEP> 6 <SEP> CH2OH <SEP> S <SEP> R <SEP> 1 <SEP> 56
<tb> Product <SEP> A <SEP> (See <SEP> above) <SEP> 0.3 <SEP> 26
<tb> Product <SEP> B <SEP> (See <SEP> above) <SEP> 1 <SEP> 27
<tb> Product <SEP> C <SEP> (See <SEP> above) <SEP> 1 <SEP> 18
<Tb>

Claims (10)

 (R), and (ii) their addition salts.  - ** C is an asymmetric carbon atom of configuration (R, S) or  asymmetric carbon configuration (R, S), (R) or (S),  - * C, in the case where R is not a hydrogen atom, is an atom of  or a CH2OH group,  R represents a hydrogen atom, an OH group or an OCH3 group;  in which

 1. Compound belonging to the family of analogs of deoxyspergualine, characterized in that it is chosen from the group consisting of (i) compounds of formula 2. Compound according to claim 1, characterized in that the asymmetric carbon ** C is of R configuration. 3. Process for the preparation of a compound of formula I or an addition salt thereof according to claim 1, said process being characterized in that it comprises the deprotection of a compound of formula:

  in which:  R represents a hydrogen atom, an OCH 3 group, an OH group,  a CH 2 OH group, an OR 'group or a CH 2 OR group,  R 'represents a group protecting the hydroxyl function,  - R1, R2, R3, identical or different, each represent a group  protective of the amine function,  - * C represents, when R is not the hydrogen atom, an atom of  asymmetric carbon configuration (R, S), (R) or (S),  - ** C represents an asymmetric carbon atom of configuration  (R, S) or (R), according to one or more reaction treatments known to those skilled in the art, to obtain the replacement of all the groups R1, R2, R3, R 'with a hydrogen atom.  wherein R, R 1, R 2, R 3, C and C retain the same meaning as above.

 wherein R 1 represents an amine protecting group, under similar operating conditions to those described for process (i) of variant A, to obtain a compound of formula II:

 wherein R, R2, R3, C and C retain the same meaning as above; and (iii) condensing the amine of formula X thus obtained with an acid of formula:

 in which R, R2, R3, R8, C and C retain the same meaning as above (ii) deprotect the compound IX thus obtained, according to a specific method for the hydrolysis of the N-R5 bond, in a solvent at room temperature and for about 1 to 5 hours, to obtain a compound of formula:

A above, to obtain a compound of formula:  (R, S) or (R), under conditions similar to those of the process of step (i) of the variant  - ** C represents an asymmetric carbon atom of configuration  present in the compound of formula VIII, and  amine protector, both different from the R5 protecting group  - R2 and R3, identical or different, each represent a group  in which:

 asymmetric carbon configuration (R, S), (R) or (S), with a compound of formula:  - * C represents, when R is not the hydrogen atom, an atom of  R5 represents a group protecting the amine function,  R 'is a protecting group of the hydroxyl function,  R represents a hydrogen atom or a CH2OR 'group in which   in which::

 (b) variant B which comprises the steps of: (i) condensing an amino acid derivative of formula  wherein R, R 1, R 2, R 3, C and C retain the same meaning as above; or

 (R, S) or (R) under conditions identical to those described in step (i) above to obtain a compound of formula II:  - ** C represents an asymmetric carbon atom of configuration  amino-protecting  - R2 and R3, identical or different, each represent a group  in which:

 wherein R1 and C retain the same meaning as in the compound of formula V, and R represents a hydrogen atom, a CH2OR 'group or an OR' group, R 'representing a hydroxyl protecting group; and, (iii) reacting a compound of formula VI obtained according to step (ii) above with a compound of formula:

 wherein R 'is a hydroxyl protecting group by the action of a dilute solution of a base in the presence of a water-miscible solvent at a temperature of about room temperature and for about 2 to 30 minutes. minutes to obtain a compound of formula:  or according to a known process when R represents a group OR '  hydrogen or a CH2OR 'group as described above,  or according to step (i) above when R represents the atom  wherein R, R 1, R 4 and C retain the same meanings as above; (ii) hydrolyzing the ester function of the compound of formula V obtained

 asymmetric carbon configuration (R, S), (R) or (S), by activation of the acid function using a carbodiimide coupling agent, in the presence of a nucleophilic agent, in an organic solvent at a temperature of between 0 and 40 ° C., and at a concentration of 1 mole of compound m for about 1 mole of compound IV, to obtain a compound of formula:  - * C, when R is not the hydrogen atom, represents an atom of  R4 represents a C1-C3 alkyl group, and,  R 'represents a group protecting the hydroxyl function,  R represents a hydrogen atom or a CH 2 OR group,  in which

 wherein R1 represents an amino protecting group, with an amino acid derivative of formula:

 (a) variant A which comprises the steps of: (i) condensing an acid of formula 4 Process according to claim 3, characterized in that it also comprises the steps chosen from the group consisting of 5. Intermediate compound, useful in the synthesis of a compound of formula I according to claim 1, characterized in that it is chosen from the group consisting of compounds of formula:

 in which:  R represents a hydrogen atom, an OCH 3 group, an OH group,  a CH 2 OH group, an OR 'group or a CH 2 OR group,  R 'represents a group protecting the hydroxyl function,  - R1, R2, R3, identical or different, each represent a group  protective of the amine function,  - * C represents, when R is not the hydrogen atom, an atom of  asymmetric carbon configuration (R, S), (R) or (S),  - ** C represents an asymmetric carbon atom of configuration  (R, S) or (R). 6. Intermediate compound, useful in the synthesis of a compound of formula I according to claim 1, characterized in that it corresponds to the formula:

 wherein R2 and R3 are the same and each represents a 1,1-dimethylethoxycarbonyl (Boc) group and ** C represents an asymmetric carbon atom of (R, S) or (R) configuration. 7. Use of an immunosuppressive substance, selected from the group consisting of the compounds of formula I and their non-toxic addition salts according to claim 1, for obtaining a medicinal product intended for therapeutic use vis-à-vis live with immune disorders. 8. Use of a substance selected from the group consisting of the compounds of formula I and their non-toxic addition salts according to claim 1, for obtaining a medicament for use in therapy with respect to Malaria screw. 9. Use of a substance selected from the group consisting of the compounds of formula I and their addition salts according to claim 1 as a pharmacological reagent. 10. Therapeutic composition characterized in that it contains, in combination with a physiologically acceptable excipient, at least one compound selected from the group consisting of the compounds of formula I and their non-toxic addition salts according to claim 1.