WO1986004901A1

WO1986004901A1 - Rennin- and acid proteases-inhibitor peptidic amino alcohol derivatives, preparation process thereof and therapeutical application thereof

Info

Publication number: WO1986004901A1
Application number: PCT/FR1986/000060
Authority: WO
Inventors: Jean Wagnon; Rémy Guegan; Colette Lacour; Dino Nisato
Original assignee: Sanofi; Institut National De La Sante Et De La Recherche M
Priority date: 1985-02-26
Filing date: 1986-02-25
Publication date: 1986-08-28
Also published as: NZ215282A; EP0214209A1; PT82077B; GR860532B; ES8900018A1; AU587426B2; AU5410086A; ES552407A0; KR860006487A; PT82077A; DK89286D0; DK89286A

Abstract

Peptidic amino alcohol derivatives which are inhibitors of rennin and of acid proteases. Said derivatives are usable in therapeutics.

Description

Amino derivative peptide inhibitors of renin and acid proteasos, the method of preparation, and their application in therapy.

The present invention relates to new peptide-inhibitor derivatives of renin and more generally acid proteases. It also relates to a process for obtaining them and applying them in therapy.

In 1970, UMEZAWA isolated, from a culture of streptomyces, a pentapeptide designated under the name of pepstatin whose structure was established later and corresponds to the formula:

Isovaleryl - L-valyl - L-valyl - Statyl - L-alanyl - Statin

in which is designated under the name of "statin" an unusual amino acid, (3S, 4S) amino-4-hydroxy-3-methyl-6 heptanoic acid.

Pepstatin has been shown to be an inhibitor of acidic proteases and acts in particular on pepsin, cathepsin D and renin. Specifically, renin, an enzyme of renal origin, is involved in the angiotensinogen sequence, angiotensin I, angiotensin II at the level of the transformation of angiotensinogen into angiotensin.

Angiotensin II being a powerful vasoconstrictor agent, intervenes in the regulation of blood pressure. Consideration has been given to using pepstatin to control high blood pressure in humans. However, since pepstatin acts on all of the acid proteases and having a low solubility in an aqueous medium and a low affinity for renin, its use in therapy has proved difficult. Pepstatin derivatives have been described in the scientific literature. For example, attempts have been made to dissolve pepstatin by lengthening the peptide chain (J. Cardiovasc. Pharmacol., 1980, 2, 687-698).

According to the present invention it has been found that it is possible to obtain very active products without lengthening the peptide chain but by introducing hydrophilic residues at different sites of a peptide analogous to pepstatin. This is all the more surprising since the prior art (patents ET 114993 and US 4,470,971) teaches that the presence of hydrophilic substituents reduces the inhibitory activity against renin.

The present invention relates to peptides having a high level of activity as inhibitors of renin and other acidic proteases.

In the present invention and in the claims the following abbreviations will be used.

Amino acids and protective or gactivating groups.

These abbreviations are in accordance with those indicated by the Nomenclature Commission of the IUPAC-IUB Biochemistry section. The most recent recommendations are reported in Eur. J. Biochem., 1984, 138, 5-7 and 9-37.

Amino Acidas:

Abu: Aminobutyric gamma acid

Ala: Alanine

Asn: Asparagine

Asp: Aspartic Acid

Cpg: Cyclopentylglycine

Gin: Glutamine Gly: Glycine

His: Histidine

Island: Isoleucine

Leu: Leucine

Met: Methionine

Nie: Norleucine

Nva: Norvaline

Phe: Phenylalanine

Phg: Phenylglycine

Ser: Serine

Val: Valine

(tauMe) His: (tele-methyl) Histidine

These amino acids are of configuration L.

Sta: Statin: 4-amino-3-hydroxy-6-methyl-heptanoic acid AHPPA: 3-aminoM hydroxy-5-phenyl pentanoic acid ACHPA: 4-amino-5-cyclohexyl-hydroxy 3-pentanoic acid

Sta, AHPPA and ACHPA are, unless otherwise indicated, in 3S, 4S configuration.

Protective groups and activators:

Ac: Acetyl

Boc: t.butyloxycarbonyl

(Boc) 2 0: bis terbutyloxycarbonic anhydride

HONSu: N-hydroxysuccinimide

OEt: Ethyl ester

OMe: Methyl ester ONp: Nitrophenyl ester ONSu: N-hydroxysuccinimide ester iVa: Isovaleryl Z: Benzyloxycarbonyl

In addition, the following abbreviations are used here:

AA: Amino acid

AcOEt: Ethyl acetate

AcOH: Acetic acid

Bop: Benzyloxy tris dimethylamino phosphonium hexafluorophosphate

TLC: Thin layer chromatography

DCCI: Dicyclohexylcarbodiimide

DCHA: Dicyclohexylamine

DCU: Dicyclohexylurea

DIPEA: Diisopropylethylamine

DMF: Dimethylform-amide

DMSO: Dimethyl sulfoxide

Ether: Ethyl ether

HOBt: 1-Hydroxy benzotriazole

MeOH: Methanol

NEM: N-ethyl morpholine

NMM: N-methyl morpholine

TA: Ambient temperature

TFA: Trifluoroacetic acid

Reduced Sta: 4-amino-6-methyl-heptane-1,3 diol.

"we debug" means: we remove the protective group Boc.

The asymmetric carbons for which the configuration * is determined are denoted C. The compounds according to the invention correspond to the following general formula:

in which :

- R1 represents an acyl group selected from alkylcarbonyl, alkoxycarbonyl, arylcarbonyl, arylalkylcarbonyl, arylalkoxycarbonyl, heterocyclylcarbonyl, heterocyclylalkylcarbonyl, in which the alkyl group is optionally substituted by hydroxy, hétérocyolylcarbonylalkylcarbonyle, hétérocyclylalcénylcarbonyle, cycloalkylcarbonyl group or a (lower alkyl) sulfonyl not substituted or substituted on the alkyl by a free amino group or carrying a protective group or by a phenyl; or a phenylsulfonyl group which is unsubstituted or substituted on the phenyl ring with lower alkyl;

- R2 represents a lower alkyl group unsubstituted or substituted by a phenyl, naphthyl, cyclohexyl, or pyridyl; - R3 represents hydrogen, lower alkenyl, phenyl, naphthyl, cycloalkyl containing 3 to 6 carbon atoms, a 5- or 6-membered monocyclic heterocyclic group unsubstituted or substituted with lower alkyl or trifluoromethyl, or alternatively a lower alkyl unsubstituted or substituted by a free amino group or carrying a protective group, by a di (lower alkyl) amino group, by a free carboxyl or esterified with lower alkyl or benzyl, by a free carbamoyl or substituted with one or two lower alkyls or with a phenyl, by a hydroxy, lower alkoxy or benzyloxy group, by a pyridylmethyloxy group, by a lower alkylthio group, (lower alkyl) suifinyl or (lower alkyl) suifonyl, by a phenyl, by a naphthyl, by a cycloalkyl containing from 3 to 6 carbon atoms or by a 5 or 6-membered monocyclic heterocyclic group unsubstituted or substituted with lower alkyl or trifluoromethyl;

- Q represents isopropyl, phenyl or cyclohexyl forming respectively with the radical

the residue of the amino acid Sta, or AHPPA or ACHPA;

- X is either a direct bond, or the residue of an amino acid such as Ala, Nva, Val, Leu, Nie, Ile, Phg, Abu, Cpg; - R4 represents hydrogen or a lower alkyl unsubstituted or substituted by an indolyl, pyridyl, imidazolyl or phenyl radical;

- R5 represents an alkyl group of 3 to 20 carbon atoms unsubstituted or substituted by a phenyl, cyclohexyl, hydroxyphenyl group, the said alkyl group containing at least one hydroxyl group and optionally an amino group, in said alkyl group all the atoms of carbon being bound to at least one hydrogen atom;

- or R4 and R5 taken together represent a 1,4 butylene or 1,5 pentylene group substituted by one or more hydroxyl or lower hydroxyalkyl groups,

with the limitation that when R4 represents hydrogen and X is other than a direct bond or one of the residues Cpg, Phg, either R1 is other than an alkylcarbonyl, alkoxycarbonyl, arylcarbonyl, aryalkylcarbonyl, arylalkoxycarbonyl or cycloalkylcarbonyl group, either R3 is other than a lower alkyl unsubstituted or substituted by a free amino group or carrying a protective group, by a carboxyl group, by a hydroxy group, by a lower alkylthio group, by a phenyl, by a naphthyl, by an imidazolyl -4; as well as their optional pharmaceutically acceptable salts with mineral or organic acids or alkali or alkaline earth metals.

The preferred products according to the present invention have the formula (I) above in which R5 contains from 1 to 3 hydroxyl groups. Particularly the products according to the present invention are of formula (I) above in which R5 represents:

a) a group -Alk-CHOH- (CH2) m-OH where

- Alk represents methylene, ethylene, a lower alkylidene group unsubstituted or substituted in the omega position by a free amino group or carrying a protective group, a benzylidene or a 2-cyclohexylethylidene group or a 2-phenylethylidene group unsubstituted or substituted on the nucleus phenyl with a hydroxyl;

Alk can not be other than methylene or ethylene when R4 is other than hydrogen;

- m is one or two;

b) a group:

- p represents an integer between zero and 5 * - R6 represents hydrogen or lower alkyl

c) a group:

p is as defined above;

- R7 represents hydrogen, a lower alkyl unsubstituted or substituted by a free amino group or carrying a protective group;

In definition (a) of R5, when R4 is other than hydrogen, Alk can only be methylene or ethylene because the products, in which Alk is other than methylene or ethylene, cannot be prepared according to the methods described .

In the definition of R1, the term alkyl designates the radicals of aliphatic hydrocarbons, saturated or unsaturated containing from 1 to 10 carbon atoms, the preferred alkyl groups, are lower alkyls as defined below.

The terms "lower alkyl", "lower alkenyl" and "lower alkylidene", as used herein, refer to radicals of saturated or unsaturated aliphatic hydrocarbons containing up to 6 carbon atoms.

The expressions "lower alkoxy" and "lower alkylthio" represent the hydroxyl and thiol groups substituted by a lower alkyl group as defined above. The term "5- or 6-membered monocyclic heterocycle" includes pyrrolidine, imidazole, thiazole, thiophene, furan, pyrrole, triazole, oxazole, isoxazole, pyridine, thiadiazoles.

By "protecting group" is meant a protecting group normally used in peptide chemistry, for example Boc, Z or iVa.

The term "acyl group" used for the definition of R1 includes residues of aliphatic, alicyclic, aromatic or heterocyclic carboxylic acids. Preferred acyl groups are the residue of esterified carbonic acid, in particular the Boc and 2 groups, the residue of alkanoic acids containing from 2 to 6 carbon atoms, in particular the iVa group, the residue of cyclohexylcarboxylic acid, residue of phenylaliphatic acids, in particular the phenylacetyl, 3-phenylpropionyl, dibenzylacetyl groups, the residue of arylcarboxylic acids, such as naphthoic acid, biphenyl carboxylic acid and unsubstituted or substituted benzoic acid on the phenyl ring, in particular the group benzoyl, the residue of a carboxylic acid whose carboxyl is linked to a 5 or 6-membered monocyclic heterocycle, in particular the picolinoyl, nicotinoyl and isonicotinoyl groups and the residue of alkanoic or alkenoic acids, such as acetic acid, propionic acid, butyric acid, valeric acid and their omega-hydroxy or omega-oxo derivatives, substituted in the omega position by a monocy heterocycle click with 5 or 6 links as illustrated above. More particularly, the present invention preferably relates to the peptide amino alcohols of formula I in which R2, R3, Q, R4 and R5 are as defined above and R1 represents one of the following groups:

NH2CH2CH2 - SO2-

BocNHCH2CH2 - SO2-

Z-NHCH2CH2 - SO2-

A - SO2 -, where A is lower alkyl

C6H5 - (CH2) a - SO2 -, where a = 0, 1 or 2

Particularly preferred are the peptide amino alcohols of formula (I) in which R2, R3, Q, R4 and R5 are as defined above and R1 represents a group chosen from

- isovaleryl,

- (pyridyl-2) -4 oxo-4 butyryl,

- (2-pyridyl) -4 4-hydroxy-butyryl,

- (pyridyl-3) -3 propionyl,

- (pyridyl-3) -4 butyryl.

- nicotinoyl,

- benzenesulf onyl. In addition, particularly preferred are the compounds of formula (I) in which R1, R2, R3, R4, R5 and Q are as defined above and R3 is such that the residue NH-CH (R3) -CO- represents the residue (tauMe) His.

The products according to the invention can be prepared according to the usual methods of peptide chemistry. Thus, according to another of its aspects, the invention refers to a process for the preparation of the peptide amino alcohol derivatives of formula I above, characterized in that an amino alcohol of formula is treated:

in which W is a protective group or hydrogen and R4 and R5 are as defined above and whose hydroxyl groups are optionally protected, with the lower alkyl ester of the amino acid of formula:

in which X 'has the definition given above for X, but X' is other than a direct bond and Q is as defined above, then step by step the elongation of the peptide chain by coupling with the following amino acids or fragments, suitably protected, the various operations being carried out using either an activated ester of the amino acid or of the fragment to be coupled, or the N-protected amino acid in the presence of DCCI, the N-protecting groups being cleaved after each coupling either by hydrogenolysis or by hydrolysis in a strong acid medium and, where appropriate, the hydroxyl groups are deprotected from the amino alcohol thus obtained by acid hydrolysis; and the product thus obtained is transformed if necessary into its pharmaceutically acceptable salts.

When the amino acid to be introduced into the sequence has a function capable of reacting in its side chain, it should be blocked by a suitable protective group which is subsequently eliminated.

The protection of the N-terminal amino acid by the group R1 is carried out according to known methods, ie before the coupling of the residue

with the next amino acid, or later.

When the compound (I) carries a substituent R3 such that the corresponding residue is an unnatural amino acid, this unnatural amino acid is prepared according to known methods. It is then coupled with the following amino acid according to the usual method.

The amino alcohol portion can be prepared by various methods. One method (method 1) consists in coupling a suitably chosen amino alcohol with the desired amino acid. When the amino alcohol chosen is not commercially available, it can be prepared by reduction of the corresponding amino acid ester. It is also possible to prepare the aminodiol derivatives 1, 2 from protected amino acids N (method 2) or to operate by condensation of an amine on an epoxy propanol or on an optically active glycerol derivative to obtain the dihydroxylated secondary amines, when R 4 is not hydrogen (method 3). To prepare the linear aminodiol derivatives 1, 3, the operation is carried out by reduction of the corresponding beta hydroxy esters (method 4). When aminodiol is present in the form of acetonide, deprotection can be carried out subsequently.

Methods 2, 3 and 4 are described below:

Method 2:

The aminodiol derivatives 1, 2 obtained from the amino acids are synthesized according to the method described in J. Chem. Soc. Chem. Comm., 1979, 875: the amino acid protected, for example by Boc or Z, is transformed into diazoketone, then treated with trifluoroacetic acid and reduced with sodium borohydride.

W represents a protective group and R represents an amino acid residue.

Diols can be obtained by this method, for example from the following protected amino acids: leucine, phenylalanine, lysine.

The chirality of alpha carbon is preserved by this process; however the reduction with sodium borohydride provides the 2 secondary epimer alcohols which can either be separated, for example by chromatography on silica gel, or used as a mixture for the subsequent reactions and separated in a subsequent stage of the synthesis.

Method 3: The diols 1, 2 derived from the N-substituted linear aminopropanediols are obtained by condensation of an amine on the racemic 1,2-propanol epoxy. The precursor aminodiol derivatives of compound (I) are then in racemic form.

R4 has the meaning given above.

To obtain an optically active aminodiol derivative, one starts with a glycerol derivative itself optically active using methods known per se. For example, the use of (R) tosyloxy-3 propa-nediol-1,2 acetonide (commercial product) provides, by aminolysis then hydrolysis in acid medium, the derivative (S) amino-3 propanediol-1, 2:

The coupling with a protected amino acid (W-AA) can be carried out on the acetonide before or after the hydrolysis; after deprotection, the aminodiol derivative of the desired configuration is obtained:

It is also possible to react (R) glycerol acetonide, the synthesis of which is described in J. Am. Chem. Soc, 1980, 102, 6304-6311.

Method 4:

The linear aminodiol derivatives 1, 3 are obtained by reduction of beta hydroxy esters with a boron hydride, such as, for example, excess sodium tetraborohydride, in a hydroxylic solvent at a temperature between 0 and 60 ° C. Dipeptides or polypeptides which contain a terminal statin in the form of an ester can also be reduced in this way. To obtain N-substituted linear aminodiol derivatives 1, 3, it is possible to first prepare the aminodiol derivatives 1, 2 of determined configuration using methods known per se, for example from an optically active derivative of glycerol such as acetonide of (R) tosyl-3 propanediol-1,2. After condensation with an amine R4NH2 and hydrolysis of the acetonide in an acid medium, the amino function is protected, for example by the action of (Boc) 20. This gives an N-protected propanediol derivative of determined configuration. The corresponding beta hydroxyester is obtained by several successive reactions: esterification of the alcohol, for example with mesyl chloride, then substitution with sodium cyanide and, after hydrolysis in an acid medium, new protection of the amine and esterification with diazomethane then deprotection in an acid medium, for example by TFA. The coupling with a protected amino acid (W-AA) or a protected polypeptide is then carried out according to the usual methods before reducing the ester to alcohol, for example by sodium borohydride.

1) R4NH2

2) H + / H20

3) (Boc) 20

D CH3SO2Cl

2) NaCN

3) H + / H2O 4) (Boc) 20

5) CH2N2

6) TFA

+ *

(R) H2NR4-CH2-CHOH-CH2-CO2CH3

1) peptide coupling

2) NaBH4 *

(R) W-AA-NR4-CH2-CHOH-CH2-CH20H

If the action of cyanide precedes the action of the amine, the final secondary alcohol is of absolute configuration

(S). The final product of formula (I) can be transformed, according to known methods, into its pharmaceutically acceptable salts with mineral or organic acids or alkali metals, preferably sodium or potassium, or alkaline earth metals, preferably calcium.

The compounds of the present invention have, like pepstatin, a marked inhibitory action on acid proteases; in particular, they have a very important inhibitory action on human plasma renin activity and, in general, clearly superior to that of the natural product: pepstatin.

The products according to the invention have been studied with regard to their therapeutic properties and in particular their enzymatic inhibitory action. More particularly, the compounds were evaluated "in vitro" on the inhibition of Human Plasma Renin activity (A.R.P.). I - EVALUATION METHOD.

The evaluation method is inspired by GUYENE (J. clin. Endocrinol. Metab., 1976, 43, 1301). The inhibition of ARP is evaluated from a pool of human plasmas, rich in renin (15 to 20 mg of angiotensin I released per milliliter and per hour), incubated for 60 minutes at 37 ° C., in a phosphate buffer at pH 7.4, in the presence of increasing concentrations of the product to be studied.

Human plasma contains the substrate: angiotensinogen and the enzyme: renin. The angiotensin I released during the reaction is measured by radioimmunoassay: Plasma Renin Activity Kit from Travenol (n ° CA 533,553). A converting enzyme inhibitor, phenylmethylsulfonyl fluoride (PMSF) is added to the incubation medium. The total incubation volume is 555 microliters distributed as follows:

- 420 microliters of human plasma

- 11 to 50 microliters of the product to be studied at variable concentrations - 119 to 80 microliters of phosphate buffer

- 5 microliters of PMSF.

A solution of acetic acid in methanol (19/1 by volume) and a solution of sodium hydroxide (IN) in methanol (2/1 by volume) are prepared. In an equivolumic mixture of these 2 solutions, a 0.001 M stock solution is prepared, of the product to be studied. Subsequent dilutions are then made in phosphate buffer. The amount of solvent present in a solution does not interfere with the results.

II - RESULTS.

The results are expressed by the dose of compound evaluated in moles which inhibits by 50% (IC50) the Human Plasmic Renin activity observed in the absence of inhibitor. The results obtained with various products of the invention are shown in the following table 1 where the IC50s of each molecule appear as regards their inhibition of the Human Plasmatic Renin Activity at pH 7.4. 5-10 doses were required to determine these IC50s. Pepstatin, as a reference substance, is always tested in parallel in each experiment. The results are expressed by their logarithmic value (-Log IC50).

TABLE 1

:: ARP inhibition:: Human: SR number: -Log IC 50:::: pH 7.4::: Pepstatin: 4.92: 42 650: 5.79: 42 336: 6.19: 42 837: 6.69: 43 009: 5.92: 43 010: 6.17: 43 093: 6.13: 43 094: 7.02: 43 095: 5.65: 43 096: 7.04: 43 195: 6 , 18: 43 251: 7.28: 43 298: 6.85: 43 378: 7.14: 43 493: 6.69: 43 494: 6.61: 43 507: 7.08: 43 533: 6, 79: 43 541: 6.88: 43 548: 6.82: 43 549: 7.76: 43 571: 7.85: 43 653: 6.95: 43 654: 5.85 43 655 6.92

43,657 6.56

43,683 7.38

43,716 6.43

The toxicity of the products according to the invention is compatible with their use in therapy.

We can therefore consider the use of the products according to the invention in therapeutic fields where the inhibition of renin-angiotansin enzyme systems is justified, in particular hypertension, peptic ulcer disease and inflammatory conditions.

Thus, the subject of the present invention is, according to another of its aspects, the pharmaceutical compositions or antihypertensive drugs containing the peptides of formula (I) or their pharmaceutically acceptable salts as active principles.

The amino peptide alcohols of the present invention can be used in therapy by injection. intravenous, intramuscular or subcutaneous. They are used in a solvent such as physiological saline (isotonic saline solution) or in a buffer such as phosphate buffer; they can also be suspended in a diluting agent, aqueous or non-aqueous, pharmaceutically acceptable. Each dosage unit can contain from 1 to 1000 mg of active ingredient.

The following non-limiting examples are given by way of illustration of the present invention. The pH of solutions in an organic solvent is measured or controlled using wet pH indicator paper.

The indicated melting points (Fc) are measured at the capillary tube.

The rotation indices (alpha D) are measured at 25 ° C.

For recording the infrared (IR) spectra, the product is dissolved in CH2Cl2 or mixed with KBr to form a pellet.

Finally, the nuclear magnetic resonance (NMR) spectra are recorded at 250 MHz in solution in DMSO, the internal standard being hexamethyldisiloxane.

The following abbreviations are used: s: singlet d: doublet m: multiplet or solid.

Moreover, . H ar means aromatic H;

. H im means H imidazolic; . H pyr means H of pyridine; . H (reduced Sta) means H of 4-amino-6 methyl-heptanediol 1,3 of 3R, 4S or 3S, 4S configuration; . approx. means approximately.

Chemical shifts (delta) are measured in ppm. EXAMPLE 1:

The .aminodiol derivative is prepared according to method 3.

1 °) (S) ((imidazolyl-4) -2 ethylamino) -3

1,2-O-isopropylidene-1,2-propanediol.

1.6 g of sodium are gradually added to 200 ml of methanol. When everything is dissolved, 6.4 g of histamine dihydrochloride in solution in hot methanol are added and the mixture is stirred for 30 minutes. The precipitate formed is filtered, the solvent is evaporated in vacuo, the residue is taken up in 100 ml of methanol, added 5 g of (R) tosyloxy-3 O-1,2-isopropylene-1,2 propanediol-1,2 and brought to reflux for 72 hours. It is concentrated under vacuum, then the residue obtained is extracted with AcOET. After evaporation of the solvent, an oil is collected which is chromatographed on a column silica. The AcOEt elutes the expected product in the form of an oil. P = 1.1 g.

NMR SPECTRUM

Delta Aspect Protons Attribution

7.46 s 1 H CH im 6.70 s 1 H CH im

1.24-1.19 2 s CH3

To a solution of Boc-Ala-OH (586 mg) in CH2Cl2 containing triethylamine (320 mg) and cooled by brine (internal temperature approximately -10 ° C.), isobutyl chloroformate (421 mg) is added and stir 45 minutes. Then added 760 mg of the preceding product, continues to stir for 3 hours at 0 ° C and allowed to return to RT 48 hours. It is taken up in water, decanted, washed with a NaHCO3 solution, dried and concentrated. An oil is obtained which is chromatographed on silica. The AcOEt-MeOH mixture at 90/10 in eluted volume, the expected product in the form of an oil.

P = 540 mg. ;

IR: (CH2Cl2); 1705-1645 cm-1.

3 °) Boc-Nle-Sta-OCH3.

In an equivolumic mixture of CH2Cl2 and dioxane (16 ml), a solution of Boc-Nle-ONSu (900 mg), HOBt (421 mg), TFA-Sta-OCH3 (0.9 equivalent) containing sufficient NMM is reacted. to bring the pH of the solution to 7. After 18 hours, evaporate to dryness, add water, extracted with ether, washed the organic phase successively with a solution of KHSO4, water, a solution of NaHCO3, then a saline solution. After drying (MgSO4) and evaporation, a purified oil is isolated by chromatography on silica gel (eluent ethyl pentaneacetate 1/1 by volume). 85% yield.

4 °) Boc-Phe-Nle-Sta-OCH3.

The Boc-Nle-Sta-OCH3 peptide (800 mg) is treated at 0 ° C. for 10 minutes in TFA (8 ml) and then the excess solvent is evaporated at RT under the vacuum of Bϋchi rotavapor. To the salt thus obtained, a solution prepared in CH2Cl2 (15 ml), Boc-Phe-ONp (924 mg), HOBt (384 mg) and NMM (400 mg), then brings the pH of the reaction medium to 7 by addition of additional NMM. After 24 hours, the mixture is evaporated to dryness, water is added, AcOEt extract, the organic phase is washed with a solution of KHSO4, several times with a solution of Na2CO3, with saline water, then dried (MgSO4). After evaporation of the solvent and filtration on silica gel (eluent pentane-ethyl acetate 4/6 by volume), the expected compound is isolated in the form of a white powder. Yield 69%.

S

5 °) Boc-Phe-Nle-Sta-OH.

The above solid (750 mg) is hydrolyzed at RT in a mixture of water (5 ml) and dioxane (10 ml) with sodium hydroxide (1.3 equivalent) for 3 hours. Evaporated to dryness with a Buchi rotary evaporator at RT, then water is added, acidified with a solution of KHSO4, extracted with 3 volumes of AcOEt, washed with salt water, dry (MgSO4). After evaporation of the solvent, a solid is isolated (yield 95%) used as such for the subsequent reactions. NMR shows the total disappearance of the peak at 3.50 ppm.

6 °) SR 43 009.

330 mg of the product obtained in step 2 are treated with 5 ml of 20% acetic acid for 3 hours at 70 ° C. It is concentrated under vacuum (pressure = 5 mm Hg), the residue is taken up in benzene and evaporated. An oil is obtained which is treated with 10 ml of TFA for 15 minutes at 0 ° C. Evaporated under vacuum, cold neutralized salt, dissolved in CH2Cl2, with a slight excess of DIPEA, then added Boc-Phe-Nle-Sta-OH (440 mg), Bop (403 mg) and stirred 48 hours at RT. Evaporated in vacuo, taken up in AcOEt, washed with a Na2CO3 solution, with a saline solution, dried and concentrated. A solid is obtained which is chromatographed on silica. The AcOEt-MeOH mixture at 70-30 by volume elutes the expected product which crystallizes on evaporation of the solvent.

P = 270 mg;

Mp = 89-92 ° C;

IR (KBr): 1690-1645 cm -1.

EXAMPLE 2:

To prepare this pure isomer, method 3 is used, starting from an optically active acetonide.

1 °) (S) Benzylamino-3 O-isopropylidene-1,2 propanediol-1,2

Benzene (150 ml) is refluxed for 72 hours, a mixture of benzylamine (7.44 g) and acetonide of (R) tosyloxy-3-propanediol-1,2 (10 g). After cooling, the precipitate is filtered, the benzene solution is washed with a dilute sodium hydroxide solution, the organic phase is dried (MgSO4) and the solvent is evaporated. The oil obtained is chromatographed on silica gel. A brown oil is eluted with the AcOEt-MeOH mixture (9/1 by volume). Yield 69%. alpha D = -3.8 ° (c = 1, MeOH).

2) (S) -benzylamino-3 propanediol-1,2.

The previously obtained compound (5.1 g) is heated at 100 ° C for 3 hours in 20% acetic acid (50 ml). After cooling, it is extracted once with ether, then basified with concentrated sodium hydroxide, and extracted several times with CH2Cl2. Dry (MgSO4). After evaporation, an oil (3.1 g) is isolated which solidifies. The CH2Cl2-isopropyl ether mixture is crystallized twice. Mp = 56-57 ° C; alpha D = -16.5 ° (c = 1, MeOH).

To a solution cooled to -10 ° C of Boc alanine (1.77 g) daas CH2Cl2 (25 ml) is added triethylamine (971 mg) then slowly isobutyl chloroformate (1.14 g) in solution in same solvent (10 ml) in 45 minutes, then adds the previous aminodiol (1.7 g) in CH2Cl2 (5 ml) and then leaves to return to RT overnight. The organic phase is washed with water, with a Na2CO3 solution, then with a KHSO4 solution and then dried over MgSO4. We chromatography on silica gel and eluted with a CH2Cl2-AcOEt mixture at 70/30 by volume. The product crystallizes from the ether-hexane mixture.

Mp = 78 ° C.

The existence of rotational isomers is visible on the NMR spectrum.

300 mg of product obtained in step 1 are deprotected by TFA. The oil obtained is taken up in 10 ml of TFA; the salt is neutralized cold with a slight excess of DIPEA then 1 ml of DMF is added and reacted with a mixture of 375 mg of Bop and 330 mg of Boc-Nle-Sta-OH obtained by hydrolysis of the corresponding ester prepared in Example 1, step 3. After 24 hours of stirring, washing with KHSO4, NaHCO3, drying and concentrating. The solid obtained is chromatographed on silica, the AcOEt-MeOH mixture at 90/10 by volume elutes the product which precipitates in hexane. Yield 320 mg. Mp = 74-75 ° C.

5 °) SR 43 094.

300 mg of the preceding product are deprotected by TFA. The solid obtained is taken up in CH2Cl2, the salt is neutralized when cold with a slight excess of NMM and then it is reacted with a mixture of 200 mg of Boc-Phe-ONp, 80 mg of HOBt and 50 mg of NMM beforehand. stirred for 2 hours in CH2Cl2. After 18 hours of stirring, washing with Na2CO3, KH SO4, drying and concentrating. The oil obtained is chromatographed on silica, the AcOEt-MeOH mixture at 90/10 by volume eluted the expected product which precipitates from ether. Yield 253 mg. Mp = 95-98 ° C.

EXAMPLE 3:

SR 43 298.

This compound is prepared using method 2 from 2,3-epoxy propanol.

1 °) (R, S) N-benzylamino-3 propanediol-1, 2 *

Mixture (R, S) C6H5CH2-NH-CH2-CHOH-CH20H.

A mixture of benzylamine (16 g) and 2,3-epoxy propanol is brought to reflux of ethanol for 48 hours. The solvent is evaporated under vacuum, then the excess benzylamine is distilled. The residue is dissolved in the minimum of dilute HCl, the impurities are eliminated with ether, then the amine (fairly highly soluble in water) is regenerated by concentrated soda, then extracted with ether . After drying (MgSO4) and evaporation, the crude product used is isolated as it is for the following reaction.

This compound is prepared according to step 3 of Example 2, by coupling with Boc-Alanine of the product previously obtained. After drying (MgSO4) and evaporation, an oil is isolated which is chromatographed on silica gel. The expected compound is eluted with the CH2Cl2 / AcOEt mixture (7/3 by volume) in the form of an oil.

3 °) SR 43 298.

This compound is prepared by operating as in Example 1 by coupling with the desired polypeptide suitably protected.

NMR SPECTRUM

NMR compares to that of SR 43 094 but many peaks are split or not resolved.

8.11-7.92-7.50 (unresolved massifs, 3 NH CO) 6.98 (d, J = 8 Hz, 1 H, NH CO2).

EXAMPLE 4:

SR 42 837

1 °) (S) (2-Phenyl) 3-ethylamino O-isopropylidene-1,2 propanediol-1,2

2) (S) (2-phenyl) 3-ethylamino-1,2-propanediol. The first 2 steps of the synthesis of SR 42 837 are carried out according to method 3, as described in Example 2, steps 1 and 2, replacing the benzylamine with phenethylamine.

To a solution of Boc-Ala-OH (1.25 g) in 20 ml of CH2Cl2, containing triethylamine (0.66 g) and cooled by brine (internal temperature approximately -10 ° C), chloroformate is added isobutyl (897 mg and stirred for 45 minutes. 1.3 g of the product obtained in the preceding step, then dissolved in a minimum of CH2Cl2, are then added, then allowed to return to RT for 18 hours. The organic phase is washed with a KHSO4 solution, NaHCO3 and ClNa, then dry and concentrate An oil is obtained which is chromatographed on silica. The pentane / ethyl acetate mixture at 20/80 by volume eluted the expected product in the form of a oil. P = 1.35 g.

4 °) SR 42 837.

250 mg of the above product is treated at 0 ° C. for 15 minutes with 8 ml of TFA. Evaporated under vacuum at 25 ° C, taken up in ether and concentrated. An oil is obtained which is taken up in acetonitrile (10 ml), cold neutralizes the salt with a slight excess of DIPEA, then adds Boc-Phe-Nle-Sta-OH (250 mg), Bop (247 mg) and shakes for 48 hours at RT. Concentrated in vacuo, taken up in AcOET, washed with a solution of NaHCO3, KHSO4, ClNa, dried and concentrated. An oil is obtained which is chromatographed on silica. The AcOEt / MeOH mixture at 90/10 by volume elutes the expected product which crystallizes from the hexane-ether mixture. P = 192 mg. Mp = 85-88 ° C.

EXAMPLE 5:

SR 43 494. 1 °) (S) (2-Phenyl) 3-ethylamino-1,2-propanediol (S) C6H5- (CH2) 2-NH-CH2-CHOH-CH20H.

This product is obtained according to Example 4, steps 1 and 2, that is to say by following method 3. Then method 4 is used to prepare the compound 43494

2 °) (S) N-Boc (2-phenylethylamino) -3 propanediol-1,2

A mixture of 7.5 g of the above product and 8.17 g of (Boc) 2 O is brought to reflux of THF (200 ml) for 3 hours, then allowed to return to RT overnight. The oil obtained is evaporated under vacuum and chromatographed on a silica column. The product is eluted by the equivolumic mixture CH2Cl2-AcOEt. P = 10.5 g.

3 °) (R) N-Boc (2-phenylethylamino) -4 3-hydroxy-butyronitrile

To a solution of the above compound (2.9 g) in pyridine (15 ml) cooled to -5 ° C is added dropwise an equivalent of freshly distilled methanesulfonyl chloride, then stored at 0 ° C for 18 hours. It is poured onto a mixture of water and ice, extracted with CH2Cl2, the organic phase is washed several times with a cold solution of 2N HCl, dried (MgSO4) and the solvent is evaporated. After evaporation, a yellow oil (3.3 g) is isolated. Stirred for 30 minutes under reflux of ethanol (50 ml) a mixture of the above compound (1.8 g) and sodium cyanide (236 mg). The solvent is evaporated in vacuo, AcOEt is added, the organic phase is washed with water, with a KHSO4 solution and then with a Na2CO3 solution. After drying (MgSO4) and evaporation, the oil obtained is chromatographed on silica gel. An oil is eluted with the CH2Cl2 / AcOEt mixture (1/1 by volume). Yield 50%. alpha D = -9.2 ° (c = 1.1, CHCl3);

IR (CH2Cl2): 2260-1690-1660 cm-1.

4) Acid (R) N-Boc (2-phenyl ethylamino) -4 3-hydroxy butyric acid

The above compound (1.0 g) in concentrated HCl (5 ml) is heated at 100 ° C. for 18 hours. It is evaporated to dryness and a solid is collected. It is redissolved in a water-dioxane mixture (1/1 by volume, 10 ml) then triethylamine is added until neutral, then stirred at RT for 18 hours in the presence of tert-butyl carbonate (787 mg) and 365 mg of additional triethylamine. It is evaporated to dryness, water is added, a few impurities are extracted with a volume of ether, the aqueous phase is acidified with a solution of KHSO4, the organic residue is extracted with ether. After drying (MgSO4) and evaporation, an oil is isolated which is purified by filtration on silica gel. Eluted with an AcOEt-MeOH mixture at 9/1 by volume. Yield 45%. alpha D = + 3.4 ° (c = 1.91, CHCl3).

5 °) (R) N-Boc (2-phenylethylamino) -4 3-hydroxy-methyl butyrate

This compound is obtained from the above acid by esterification with diazomethane in ether. After purification by filtration on silica gel using a 2/3 by volume pentane-ether mixture as eluent, an oil is obtained. alpha D = + 5.7 ° (c = 2.7; CHCl3);

IR (CH2Cl2): 1735-1690-1665 cm-1.

The above compound (270 mg) is dissolved in TFA at 0 ° C for 15 minutes. The excess acid is evaporated at RT under vacuum. The salt obtained, neutralized with NMM, is added in the form of a suspension in THF to a solution of the activated ester of Boc alanine (166 mg) prepared according to example 1, step 2, replacing CH2Cl2 with THF and triethylamine by NMM. After contact for 18 hours at 0 ° C., it is evaporated to dryness at RT, water and AcOEt are added, the solution is washed with a solution of KHSO4, with water, then with a solution of NaHCO3 and a saline solution. . After drying (MgSO4) and evaporation, the residue is chromatographed on silica gel. The expected compound is eluted, in the form of an oil, with the mixture of pentane and ethyl acetate (2/3 by volume).

IR (CH2Cl2): 3430-1730-1710-1655 cm-1.

The preceding compound (220 mg) is dissolved for 15 minutes by TFA at 0 ° C. After cold evaporation of the excess acid on a Buchi rotary evaporator, the residue is dissolved in CH2Cl2 and neutralized under cold conditions with DIPEA. A solution prepared from Boc-Phe-Nle-Sta-OH (311 mg), Bop (257 mg) containing diisopropylamine in CH2Cl2 is then added and the mixture is stirred for 18 hours at RT. The organic solution is then washed with water, with a KHSO4 solution, with a KHCO3 solution, the organic phase is dried (MgSO4), the solvent is evaporated and the residue is chromatographed on silica gel (eluent: AcOEt). The compound crystallizes on evaporation of the solvent. Mp = 82-83 ° C.

8 °) SR 43 494.

This product is prepared from the ester obtained in the previous step, operating according to method 4. To the ester (67 mg) in 100% ethanol (5 ml), solid NaBH4 (12 mg) is added. and stirred at RT for 5 hours. Acidified with a solution of KHSO4 to a pH close to 4, then evaporated to dryness. Water is added, then extracted with AcOEt, the organic phase is washed with water and carbonated water, then dried (MgSO4). After evaporation, the residue is chromatographed on silica gel (eluent AcOEt / MeOH at 9/1 by volume).

NMR SPECTRUM:

This compound differs from its precursor the ester SR 43 762 by the disappearance of the 2 OCH3 peaks at 3.53 and 3.51 ppm, the disappearance of a multiplet between 2.5 and 2.0 ppm attributed to the group CH2CO2CH3 and the appearance of an additional CH2 group (CH2-CH2-OH) between 1.70 and 1.0 ppm (23 H); 1.23 ppm, s, (CH3) 3C; 1.06 ppm, d, J = 7 Hz, CH3 (Ala).

EXAMPLE 6

SR 43 378.

The 1,2-aminodiol derivative is prepared according to method 3.

1 °) Boc-Nle-AHPPA-OCH3.

Boc AHPPA OCH3 (3.0 g) is dissolved in TFA (20 ml) according to the usual technique, the salt obtained is dissolved in CH2Cl2 then neutralized at 0 ° C with NMM. Then added to the reaction medium Boc-Nle-ONSu (3.3 g), HOBt (1.6 g) and then stirred for 18 hours at RT while maintaining the pH of the solution around 7-8 by addition of additional NMM. After the usual treatments, the residue is chromatographed on silica gel (eluent CH2Cl2-AcOEt, 1/1 by volume). The peptide crystallizes on evaporation of the solvent on a cold Bϋchi rotary evaporator (yield 80%). Mp = 89-90 ° C;

IR (CH2Cl2): 3420-1715-1675 cm-1.

2 °) Boc-Phe-Nle-AHPPA-OCH3.

The above compound (3.5 g) is dissolved at 0 ° C in TFA (30 ml). The salt obtained by evaporation of the excess reagent according to the usual method is dissolved in

CH2Cl2 and neutralized at 0 ° C with NMM then added at 0 ° C to a previously obtained solution of Boc-Phe-ONp (3.3 g), HOBt (1.33 g) and NMM (808 mg) in CH2Cl2. The mixture is stirred for 18 hours at RT then the organic phase is washed with water, with a solution of Na2CO3, with a solution of KHSO4, with water, then dried (MgSO4). By evaporation of the solvent, a solid is isolated which is triturated in ether, then filtered; homogeneous in CCM. Yield 75%. Mp = 182-183 ° C.

3 °) Boc-Phe-Nle-AHPPA-OH.

The ester described in the previous step (2 g) in solution in dioxane (100 ml), is hydrolyzed at RT for 4 hours with sodium hydroxide (178 mg) dissolved in water (10 ml). The subsequent treatment according to Example 10, step 2, provides the acid which is triturated in isopropyl ether, filtered and dried under vacuum (yield 95%). Mp = 161-162 ° C.

4) SR 43 378.

Condensation of the previous acid with lee

prepared in Example 2, step 3, provides the expected peptide which is purified by chromatography on silica gel, eluting with the AcOEt-MeOH mixture at 4/1 by volume.

EXAMPLE 7:

(3S, 4S) N - ((pyridyl-2) -4 oxo-4 butyryl) -Phe-

SR 43195.

This compound is prepared by reducing Boc-Val-Sta-OCH3. NaBH4 (160 mg) is added to 100% ethanol (8 ml) and then, after dissolution, the solid Boc-Val-Sta-OCH3 dipeptide (370 mg is added and stirred for 270 minutes at RT, then brought to 45 ° C. for 30 minutes The reaction is followed by TLC on silica plates, water is then added (1 vol) and then acidified with care by a solution of KHSO4, the ethanol is evaporated under vacuum at 40 °. C and extracted with 4 volumes of AcOEt, washed with water, saline and dry water (MgSO4) After chromatography on silica gel (eluent AcOEt-MeOH at 9/1 by volume) a homogeneous product is eluted in CCM.

Yield 77%; alpha D = -33 ° (c = 0.76; MeOH);

Mp = 65-67 ° C (triturated in the ether-hexane mixture).

One can also first prepare (3S, 4S) N-Boc 4-amino-6-methyl-1,3-heptanediol and then couple it with valine according to conventional methods. The preparation of the aminodiol derivative is then carried out as follows:

To a solution of NaBH4 (261 mg) in 100% ethanol (15 ml) the Boc-Sta-OMe (500 mg) is added and the mixture is stirred for 3 hours at RT. Water is added, then a KHSO4 solution, the ethanol is evaporated in vacuo (bath temperature around 35 ° C). Extraction is carried out with AcOEt, washed with water, saline water, dry (MgSO4). The residue is chromatographed on silica gel. Eluted by the mixture of pentane and ethyl acetate (1/1 by volume) and crystallizes from a mixture of ether and hexane.

Yield 60%;

Mp = 68-70 ° C.

This product is prepared according to the usual coupling methods.

3 °) SR 43195.

The product prepared in the previous step (220 mg) is treated with TFA for 20 minutes at 0 ° C. The medium is evaporated to dryness and then the trifluoracetate is dissolved in acetonitrile (25 ml). We neutralize by adding

DIPEA then added (2-pyridyl) -4 oxo-4 butyric acid (70 mg) and Bop (170 mg). It is checked that the pH is greater than 7 and then stirred for 48 hours at RT. The medium is evaporated, taken up in AcOEt and then washed successively with solutions of KHSO4, NaHCO3 and ClNa. After drying and concentration, chromatography on silica. The AcOEt-MeOH mixture at 9/1 by volume elutes the amorphous product which precipitates in AcOEt; P = 101 mg.

EXAMPLE 8:

(3S, 4S) N - ((pyridyl-2) -4 hydroxy-4 butyryl)

SR 43251.

The SR 43195 prepared in Example 7 (53 mg) is solubilized in MeOH (20 ml) in the presence of NaBH4 (25 mg). We keep one night at TA. KHSO4 is added up to pH 3, then NaHCO3 up to pH 8. MeOH is evaporated, then the medium is taken up in water and then in AcOEt. The organic phase is washed with a saturated ClNa solution, dried and concentrated. The residue is dissolved in a little MeOH and then filtered. Evaporate MeOH with a nitrogen jet and then precipitate the expected product in ether; P = 45 mg. 85% yield.

EXAMPLE 9:

SR 43095. 1) (S) (2-Phenyl) 3-ethylamino-1,2-propanediol prepared in Example 4, step 2.

To a solution of Boc-Nle-OH (1.18 g) in 25 ml of THF containing 515 mg of NMM and cooled by brine (internal temperature: approximately -10 ° C.), chloroformate of dropwise is added. isobutyl (700 mg) and stir 45 minutes. 1 g of (phenyl-2) ethylamino-3-propanediol-1,2 dissolved in the minimum of THF is then added and the mixture is returned to RT for 18 hours. Concentrated in vacuo, taken up in AcOEt, washed with a solution of NaHCO3, a solution of KHSO4, dried and concentrated. An oil is obtained which is chromatographed on a silica column, eluting with an equivolumic mixture CH2Cl-AcOEt. 1.6 g of the expected product are collected in the form of an oil.

408 mg of the product obtained previously is deboculated according to the usual method with 10 ml of TFA. An oil is obtained which is taken up in CH2Cl2 (10 ml). The salt is cold neutralized by a slight excess of DIPEA and reacted with a mixture of Bop (442 mg) and

Boc-Nle-Sta-OH (388 mg). After 18 hours of stirring at RT, washing with KHSO4, with NaHCO3 and with water then drying and concentrating. A solid is obtained which is chromatographed on silica; the AcOEt-MeOH mixture at 9/1 by volume eluted the expected product which crystallizes from pentane; P = 400 mg. Mp = 70-72 ° C.

4 °) SR 43095.

160 mg of the product obtained above are debugged according to the usual method. An oil is obtained which is taken up in CH2Cl2, then the salt is cold neutralized with a slight excess of NMM, Boc-Nle-ONSu (83 mg), HOBt (39 mg) is added and the mixture is stirred for 18 hours at RT. Wash with water, par. KHSO4, NaHCO3, dries and concentrates. An oil is obtained which is chromatographed on silica. The AcOEt-MeOH mixture at 9 / l by volume elutes the expected product which precipitates from hexane; P = 110 mg. Mp = 97-100 ° C.

EXAMPLE 10:

SR 43096

prepared in Example 9, step 3.

2 °) SR 43096.

The product obtained previously is debugged according to the usual method by TFA. An oil is obtained which is dissolved in CH2Cl2; the salt is neutralized cold with a slight excess of NMM, then reacted with a mixture of Boc-Phe-ONp (127 mg), HOBt (50 mg) and NMM, previously stirred for 2 hours in CH2Cl2. After 18 hours of stirring, the mixture is washed with water, with NaHCO3, with KHSO4, dried and concentrated. An oil is obtained which is chromatographed on silica. The AcOEt-MeOH mixture at 9 / l by volume elutes the expected product which precipitates from pentane; P = 110 mg. Mp = 94-95 ° C.

EXAMPLE 11:

SR 43010 1 °) (S). ((Indolyl-3) -2) 3-ethylamino-1,2-prcpanediol

This compound is prepared by following method 3 as described in Example 2, steps 1 and 2.

This compound is prepared by following the method described in Example 2, step 3. alpha D = -1.2 ° (c = 1; CH2Cl2).

This compound is obtained by coupling with Boc-Nle-Sta-OH according to the usual method. Mp = 86-88 ° C.

4 °) SR 43010. This compound is prepared by coupling with Boc-Phe-ONp according to the usual methods. Mp = 98-100 ° C.

EXAMPLE 12:

(3S, 4S) N- ((pyridyl-2) -4 hydroxy-4 butyryl) -

SR 43 541. 1 °) (3S, 4S) N-Boc 4-amino-6-methyl-1,3-heptanediol

To a solution of NaBH4 (261 mg) in 100% ethanol (15 ml), Boc-Sta-OMe (500 mg) is added and the mixture is stirred for 3 hours at RT. Water is added, then a KHSO4 solution, the ethanol is evaporated in vacuo (bath temperature around 35 ° C). Extraction is carried out with AcOEt, washed with water, saline water, dry (MgSO4). The residue is chromatographed on silica gel. Eluted with a mixture of pentane and ethyl acetate (1/1 by volume) and crystallized from a mixture of ether and hexane. Yield 60%.

Mp = 68-70 ° C.

The preceding compound (1.3 g) is treated at 0 ° C with TFA (12 ml) for 15 minutes. The salt obtained after evaporation plus CH2Cl2 is neutralized by a slight excess of NMM, and a CH2Cl2 solution of Boc-Ala (ONSu) (1.5 g) and HOBt (800 mg) is added. The mixture is stirred at RT for 48 hours while maintaining the pH around 7 by addition of additional NMM. The medium is then washed with a solution of NaHCO3 then with a solution of KHSO4, water, then dried (MgSO4) and evaporated. The residue is chromatographed on silica gel (eluent AcOEt-MeOH, 8/2 by volume).

This aminodiol derivative can also be prepared directly by reduction of Boc-Ala-Sta-OCH3. NaBH4 (160 mg) is added to 100% ethanol (8 ml) and then, after dissolution, the solid Boc-Ala-Sta-OCH3 dipeptide (370 mg) is added and stirred for 270 minutes at RT, then brought to 45 ° C for 30 minutes. The reaction is monitored by TLC on silica plates. Then add water (1 vol) then carefully acidify with a solution of KHSO4, evaporate the ethanol under vacuum at 40 ° C and extract with 4 volumes of AcOEt, wash with water, saline and dry (MgSO4). After chromatography on silica gel (eluent AcOEt-MeOH at 9/1 by volume), a homogeneous product in TLC is eluted. Yield 77%).

IR (CH2Cl2): 1710 and 1672 cm-1; alpha D = -60.8 ° (c = 0.72; MeOH).

3 °) N - ((pyridyl-2) -4 oxo-4 butyryl) -Phe-His-Sta-OMe

This compound is prepared from

Boc-Phe-His-Sta-OCH3 according to the method described in Example 7, step 4.

4 °) N- ((2-pyridyl) -4 4-hydroxy-butyryl) -Phe-His-Sta-OMe.

This compound is obtained by reduction of the previous one with NaBH4 according to the method described in Example 8.

5 °) SR 42 541.

The compound prepared in step 4 is hydrolyzed and then coupled after acidification of the salt of the carboxylic acid with the diol prepared in step 2, to obtain the expected product.

EXAMPLE 13:

SR 43 507. 1 °) Benzylamino-3 propanol. The methyl ester of 3-benzylaminopropanoic acid (5.0 g) is dissolved in 150 ml of anhydrous ether and cooled by an ice bath, then the double lithium aluminum hydride is added in portions ( 1.3 g) and stirred for 2 hours at RT. The excess hydride is destroyed by successively adding water (1.3 ml) a 15% sodium hydroxide solution (1.3 ml) and then water (4 ml). The solid is filtered, the ethereal phase is evaporated, a yellow oil is isolated.

A solution of 1.15 mg of Boc-Ala-OH in CH2Cl2 is prepared to which 819 mg of isobutyl chloroformate in 5 ml of CH2Cl2 is added dropwise, then the product obtained in step is added after a few minutes previous and let return to TA overnight. Poured into water, washed with a solution of Na2CO3, with water, a solution of KHSO4, dried (MgSO4) and concentrated. The residue is chromatographed on silica, eluting with an equivolumic mixture CH2Cl2 / AcOEt.

3 °) SR 43 507

This compound is prepared following the usual methods.

Using the same preparation methods indicated in (), the compounds described in Table 2 were obtained. They are characterized by their NMR spectrum.

NMR SPECTRUM

Claims

CLAIMS.

1. Aminoalcohol peptide derivative of formula:

in which :

- R1 represents an acyl group chosen from alkylcarbonyl, alkoxycarbonyl, arylcarbonyl, arylalkylcarbonyl, arylalkoxycarbonyl, heterocyclylcarbonyl, heterocyclylalkvlcarbonvle, in which the alkyl group is optionally substituted by a hydroxy, heterocycylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbonylcarbyl group substituted or substituted on the alkyl by a free amino group or carrying a protective group or by a phenyl; or an unsubstituted phenylsulfonyl group having been substituted on the phenyl ring by lower alkyl;

- R2 represents a lower alkyl group unsubstituted or substituted by a phenyl, naphthyl, cyclohexyl, or pyridyl; - R3 represents hydrogen, lower alkenyl, phenyl, naphthyl, cycloalkyl containing 3 to 6 carbon atoms, a 5- or 6-membered monocyclic heterocyclic group unsubstituted or substituted with lower alkyl or trifluoromethyl, or alternatively a lower alkyl unsubstituted or substituted by a free amino group or carrying a protective group, by a di (lower alkyl) amino group, by a free carboxyl or esterified with lower alkyl or benzyl, by a free carbamoyl or substituted with one or two lower alkyls or with a phenyl, by a hydroxy, lower alkoxy or benzyloxy group, by a pyridylmethyloxy group, by a lower alkylthio group, (lower alkyl) sulfinyl or (lower alkyl) sulfonyl, by a phenyl, by a naphthyl, by a cycloalkyl containing from 3 to 6 carbon atoms or by a 5 or 6-membered monocyclic heterocyclic group unsubstituted or substituted with lower alkyl or trifluoromethyl;

- R4 represents hydrogen or a lower alkyl unsubstituted or substituted by an indolyl, pyridyl, imidazolyl or phenyl radical;

- R5 represents an alkyl group of 3 to 20 carbon atoms unsubstituted or substituted by a phenyl, cyclohexyl, hydroxyphenyl group, the said alkyl group containing at least one hydroxyl group and optionally an amino group. - or R4 and R5 taken together represent a 1,4 butylene or 1,5 pentylene group substituted with at least 1 hydroxyl or lower hydroxyalkyl group;

the residue of the amino acid Sta, or AHPPA or ACHPA;

- X is either a direct bond, or the residue of an amino acid such as Ala, Nva, Val, Leu, Nie, Ile, Phg, Abu, Cpg:

with the limitation that when R4 represents hydrogen and X is other than a direct bond or one of the residues Cpg, Phg, either R1 is other than an alkylcarbonyl, alkoxycarbonyl, arylcarbσnyle, arylalkylcarbonyl, arylalkoxycarbonyl or cycloalkylcarbonyl group, either R3 is other than a lower alkyl unsubstituted or substituted by a free amino group or bearing a protective group, by a carboxyl group, by a hydroxy group, by a lower alkylthio group, by a phenyl, by a naphthyl, by an imidazolyl -4;

as well as any pharmaceutically acceptable salts of said amino peptide alcohol with mineral or organic acids or with alkali or alkaline earth metals.

2. A peptide amino alcohol derivative according to claim 1, characterized in that R5 contains from 1 to 3 hydroxyl groups.

3. Aminoalcohol peptide derivative, according to claim 1, of formula:

-CHOH- (CH2) m-OH

in which R1, R2, R3, Q, X and R4 are as defined in claim 1,

- Alk represents methylene, ethylene, a lower alkylidene group unsubstituted or substituted in the omega position by a free amino group or carrying a protective group, a benzylidene or a group

2-cyclohexylethylidene or a 2-phenylethylidene group unsubstituted or substituted on the phenyl ring by a hydroxyl; Alk can not be other than methylene or ethylene when R4 is other than hydrogen;

- m is one or two.

4. Aminoalcohol peptide derivative, according to claim 1, of formula:

in which R1, R2, R3, R4, Q and X are as in claim 1 and:

- p represents an integer between zero and five; - R6 represents hydrogen or lower alkyl.

Amino alcohol derivative, according to claim 1, of formula:

in which R1, R2, R3, Q, X and R4 are as defined in claim 1,

- p represents an integer between zero and 5;

- R7 represents hydrogen or a lower alkyl unsubstituted or substituted by a free amino group or carrying a protective group;

6. Aminoalcohol peptide derivative according to one of claims 1 to 5 characterized in that R1 represents one of the following groups:

NH2CH2CH2 - SO2-

BOCNHCH2CH2 - SO2-

Z NHCH2CH2 - SO2- A - SO2 -, where A is lower alkyl

C6H5 - (CH2) a - SO2 -, where a = 0, 1 or 2

or one of the optional salts of mineral or organic acids which are pharmaceutically acceptable of the said amino peptide alcohols.

7. aminoalcohol peptide derivative according to one of claims 1 to 5, characterized in that R1 represents a group chosen by the following groups

- isovaleryl _ (pyridyl-2) -4 oxo-4 butyryl

- (pyridyl-2) -4 hydroxy-4 butyryl

- (pyridyl-3) -3 propionyl

- (pyridyl-3) -4 butyryl

- nicotinoyl - benzene sulfonyl.

8. Aminoalcohol peptide derivative according to any one of claims 1 to 7 characterized in that R3 is such that the residue -NH-CH (R3) -CO- represents the residue (tauMe) His. 9. aminoalcohol peptide derivative according to any one of claims 1 to 8, characterized in that R ₄ represents a lower alkyl unsubstituted or substituted by an indolyl, pyridyl, imidazolyl or phenyl radical; and X represents the residue of an amino acid such as Ala, Nva, Leu, Nie, Ile, Abu, Val.

10. Process for the preparation of the amino acid peptide derivatives according to claim 1, characterized in that an amino alcohol of the formula is treated:

in which W is a protective group or hydrogen and R4, R5 are as defined above and whose hydroxyl groups are optionally protected, with the lower alkyl ester of the amino acid of formula

in which X 'has the definition given above for X, but X' is other than a direct bond and Q is as defined above, then we proceed, step by step, to the elongation of the peptide chain by coupling with the following amino acids or fragments, suitably protected, the various operations being carried out using either an activated ester of the amino acid or of the fragment to be coupled, or the N-protected amino acid in the presence of DCCI, the N groups - protectors being cleaved after each coupling either by hydrogenolysis or by hydrolysis in a strong acid medium and, where appropriate, the hydroxyl groups are deprotected from the amino alcohol thus obtained by acid hydrolysis; and the product thus obtained is transformed, if necessary, into one of its salts of mineral or organic acids or of alkali or alkaline earth metals.

11. Pharmaceutical compositions characterized in that they contain a product according to one of claims 1 to 9 as an active ingredient.

12. Pharmaceutical compositions according to claim 9, usable in particular for the treatment of arterial pressure, characterized in that they contain from 1 to 1000 mg of product according to claim 1 per dosage unit mixed with a pharmaceutical excipient.