WO2014064162A1

WO2014064162A1 - Neprilysin inhibitors

Info

Publication number: WO2014064162A1
Application number: PCT/EP2013/072192
Authority: WO
Inventors: Bernard Pierre Roques; Marie-Claude Fournie-Zaluski; Hervé Poras
Original assignee: Pharmaleads
Priority date: 2012-10-23
Filing date: 2013-10-23
Publication date: 2014-05-01
Also published as: FR2997080B1; FR2997080A1

Abstract

Neprilysin inhibitors are described. The compounds or compositions containing them can be used as antidiarrheal agents, antisecretory agents, anxiolytic agents, antidepressants, cicatrizants, fluid effusion reducers or in the treatment of cardiac fluid overload, oedema and glaucoma.

Description

Inhibitors of Nepriysin

FIELD OF THE INVENTION

The present invention relates to inhibitors of nepriysin.

BACKGROUND OF THE INVENTION

Metailoproteinases represent an important group of hydrolases involved in many physiological regulatory processes by being responsible for either maturation (formation of an active molecule from an inactive precursor) or inactivation (formation of inactive metabolites at from an active molecule) of peptides or proteins. Among these regulatory enzymes, nepriysin or neutral endopeptidase (NEP) plays an important role. Indeed, in mammals, it is widely distributed throughout the body (Roques et al (1993) Pharmacol Rev., 45, 87-146); it is particularly abundant in the brush border of the kidney and the intestine. It is also found in the central nervous system, in the bones, dermis, muscles and organs. This wide tissue distribution suggests the possible existence of several substrates. Thus, for example, nepriysin by degrading enkephalins is involved in the control of their physiological functions, particularly pain and mood (Roques et al., (2012) Nature Rev. Drug Discov 11, 292-311). Indeed, the protection of enkephalins (endogenous opioids having the same affinity as morphine for opioid receptors) of the action of NEP increases their extracellular concentration and leads to analgesic, antidepressant and anxiolytic effects (Roques et al., 2012). Nature Rev. Drug Discov 11, 292-311, Noble et al (2008) Psychopharmacology 196, 327-335, Komg et al (1996) Nature 383, 535-538). By degrading plasma atrial natriuretic peptide and endothelin, it is likely to correct some forms of cardiovascular disorders and regulate water balance in the body. At the intestinal level, the protection of enkephalins from the action of NEP has anti-secretory and anti-diarrheal effects illustrated by the NEP inhibitor thiorphan (Roques et al., (1980) Nature 288, 286-288). in the human clinic since 1992. Moreover, NEP has recently been identified in the dermis (Morisaki et al., (2010) J Biol Chem 285, 39819-39827) where it plays a negative role in cell renewal (cicatrization). and wrinkle formation (Tsukahara et al (2001) J. Invest, Dermatol 117, 671-677). NEP inhibitors are therefore likely to be used in dermatology and cosmetology. Finally, the inhibition of NEP is likely to eliminate various fluidic effusions (joints, pleura, glaucoma, ascites) by protection of peptides such as natriuretic peptides. This enzyme therefore represents a particularly advantageous therapeutic target for improving various pathologies.

Neprilysin (NEP, EC3.4.24.11) is a zinc-metalloprotease of the M13 family of Glu- zincins (clan MA (E)). It was cloned and sequenced (Devault et al (1987) EMBO J. 6, 1317-322) and its three-dimensional structure was obtained by X-ray crystallography (Oefner et al., (2000) J.Mol.Biol. , 341-349). It contains the two consensus sequences, characteristics of this family. They correspond to sequences ⁵⁸³ HEITH ⁵⁸⁷ and ⁶⁴⁶ ENIAD ⁶⁵⁰ , ⁵⁸³ H, ⁵⁸⁷ H and ⁶⁴⁶ E being zinc ligands and ⁵⁸⁴ E being catalytic glutamate.

The classical strategy for developing effective inhibitors of this type of enzyme consists in designing a molecule recognizing the different protease binding sub-sites and having a group having a high affinity for zinc.

The best zinc ligands used in the design of the main inhibitors described in the literature are: -SH or -S ^" thiols, which are monovalent ligands, -COOH or -COO ^" carboxylates which can act as monodentates or bidentates phosphonate or phosphinate groups: -P (O) (OH) ₂ or -P (O) OH, which may also be mono- or bidentates, hydroxamates -CO-NHOH or N-formylhydroxylamines -NH (OH) C (O) H which behave as bidentates (for review see Thorsett ED and Wyvratt MJ in "Neuropeptides and their peptidases" AJ Turner Ed., Ellis Horwood series in Biomedecine, 1987, pp 229-292).

Many NEP inhibitors, having one of the zinc ligands described above, carried by a peptidomimetic backbone have been synthesized (Roques et al., (1980) Nature 288, 286-288, Fournié-Zaluski et al (1983) J. Chem., 26-60-65; Supplied-Zaluski et al. (1984), Pharmacol 102, 525-528; Chipkin (1986) Future Drugs 11, 593-606; Hernandez et al. (1988). J.Med.Chem 31, 1825-1831, Seymour et al (1989) Hypertension 14, 87-97, Northridge et al (1989) Lancet 2, 591-593, Chen et al (1998) PNAS 95 , 12028-12033, Maw et al (2006) Biol Chem., DrugDes 67, 74-77, Glossop et al (2011) Biochem Med Chem 21, 3404-3406). Some have been co-crystallized with Neprilysin (Oefner et al (2004) Acta Crystallogr., Sect D, 60, 392-396).

Virtually all the inhibitors described above have a peptide unit and thus one or more amide link (s). For example, peptidomimetic derivatives comprising at least one amide bond and further comprising a disulfide unit have been described in the articles Noble et al. 1992 (Journal of Pharmacology and Experimental Therapeutics 261). (1992), 1, 181-190), Noble et al. 1997 (Pain 73 (97), 383-391), and in the patent documents WO 2009/138436 and FR 2892413. Moreover, they generally possess a molecular weight (PM ) greater than 500 Da. These characteristics are unfavorable for the crossing of the physiological, blood-brain, dermal and intestinal barriers, for example, and as a result these inhibitors have a rather low oral bioavailability.

The objective of the present invention is therefore to provide neprilysine inhibitors overcoming the disadvantages described above. BRIEF DESCRIPTION OF THE INVENTION

The invention relates to compounds having the following general formula (I):

RS-C (RiR ₂ ) -CO-CH ₂ -CH (R ₃ ) -CO-R 4 (I)

with R, R 1, R ₂ , R ₃ and R ₄ as defined in claim 1.

The invention also relates to pharmaceutical or cosmetic compositions comprising at least one compound of the present invention.

The invention also relates to the compounds of the present invention or pharmaceutical compositions containing them for their use as antidiarrheal, antisecretory, anxiolytic, antidepressant, cicatrisant, reducer of watery effusions or in the treatment of cardiac water overload, edema and glaucoma.

DEFINITION

Alkyl groups denote linear or branched hydrocarbon chains of Cl, C2, C3, C4, C5 or C6 alkyl, particularly methyl, ethyl, w-propyl, ^z-propyl, n-butyl, i-butyl or / - butyl.

Examples of aromatic or saturated 5- or 6-membered heterocycles comprising at least one sulfur, oxygen or nitrogen atom include the following radicals: thienyl, pyrrolyl, imidazoyl, pyrazolyl, isothiazolyl, pyridyl, pyrazinyl, pyrimidinyl pyridazinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrazolinyl, piperidyl, piperazinyl, thiadiazolyl, furyl, pyranyl, isoxazolyl, morpholinyl, furazanyl, oxazolyl, oxazolidinyl and oxazolinyl.

The term "halogen" as used herein refers to chlorine, bromine, iodine or fluorine. DETAILED DESCRIPTION OF THE INVENTION

The compounds of the present invention are characterized by: i) an original thio-ketone unit capable of interacting with the zinc of NEP in a mono or bidentate manner, ii) an absence of peptide motif (thus lacking a friendly link of), iii) a backbone with a minimum number of groups capable of interacting with the subsites of the enzyme, thus making it possible to reach a nanomolar affinity for NEP, iv) a low molecular weight.

In addition, with the aim of further improving the bioavailability, the solubility, and the pharmacokinetic parameters of these compounds, labile protections on the functional groups of the described inhibitors, of the "prodrug" type, introduced on one or both of the terminal parts, are also described. This is also the case of homodization by formation of a disulfide bridge, known to improve the passage of endothelial membranes (Fournié-Zaluski et al (1992) J. Med Chem, 35, 2474-2481). and thus further enhance the oral bioavailability and crossing of physiological barriers.

The compounds of the present invention may be administered parenterally, topically, nasally or orally.

The compounds of the present invention have the following general formula (I):

RS-C (RIR ₂₎ -CO-CH2-CH (R ₃₎ -CO-R4 (I)

in which:

a) R represents:

a hydrogen;

an alkanoyl group R'CO-, with R 'representing a linear or branched alkyl group of 1 to 6 carbon atoms, which may or may not be substituted by one or more halogen atoms;

an alkylthio group R'S- or alkanoylthio R'COS-, with R 'having the same definition as above;

an alkoxyalkyloxycarbonyl R "C (O) O-CH (R" ') OC (O) - with

representing, independently of one another, an alkyf group

branched, from 1 to 6 carbon atoms;

an S-cysteinyl (H ₂ N) (CO ₂ H) CH-CH ₂ -S- group; or

a group ~ SC (RiR ₂ ) -CO-CH ₂ -CH (R ₃ ) -CO-R 4, with R 1, R ₂ , R ₃ and R ₄ as below forming a compound of formula (I) symmetrical; i represents

a linear or branched alkyl group of 1 to 6 carbon atoms, substituted or not by:

^H a -OR ₅ , -SR ₅ or -SO-R _{5 group} , with R ₅ representing a hydrogen, a linear or branched alkyl group of 1 to 4 carbons, a phenyl or benzyl group;

^■ a -C0 ₂ R 6 group, with R ₆ representing a hydrogen, an alkyl group, linear or branched, comprising 2 to 4 carbon atoms or a benzyl group;

^B a -NR7R8 group, with R ₇ and R _$ representing, independently of one another, a hydrogen, a linear or branched alkyl group of 1 to 4 carbon atoms, a phenyl or benzyl group or with -NR ₇ Rg taken together representing a saturated 5- or 6-membered heterocycle comprising one or more heteroatoms selected from oxygen and nitrogen, preferably representing a morpholine or piperidine;

^■ a carboxamide group -CONR ₇ R 8, with -NR7R8 having the same definition as above;

^■ phenyl, substituted or not by one or more halogens selected from fluorine or bromine, or by a - OR5, with R5 having the same definition as above;

^■ an aromatic 5- or 6-membered ring comprising 1 or 2 heteroatoms chosen (s) selected from oxygen, nitrogen and sulfur;

⁸ saturated cyclic compound of 5 or 6 membered heterocyclic or saturated 5- or 6-membered ring comprising 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur;

a 5- or 6-membered saturated 5- or 6-membered saturated or saturated heterocyclic ring compound having 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur;

a phenyl group, substituted or not, with one or more halogens selected from fluorine or bromine, or with a group-ORs, with R5 having the same definition as above;

and R2 is hydrogen; or R 1 and R ₂ are identical and represent a linear or branched alkyl group of 1 to 6 carbon atoms; or

-C (RiR ₂ ) - taken together represents:

^B a 5-membered saturated cyclic compound, attached or not to an aromatic ring, preferably leading to an indanyl ring;

⁸ a cyclic saturated compound to 6-membered ring;

^{B is} a 6-membered saturated heterocyclic compound having 1 heteroatom, at the 4-position, selected from oxygen, nitrogen and sulfur, wherein when the heteroatom is nitrogen, the nitrogen is substituted or unsubstituted by an alkyl group of 1 to 6 carbon atoms, an alkanoyl group, a phenyl group or a benzyl group;

3 represents:

a linear or branched alkyl group of 1 to 6 carbon atoms, substituted or not by:

^B a group -OR ₅ or -SR ₅ or -S (O) Rs, with R 5 representing a hydrogen, a linear or branched alkyl group of 1 to 4 carbons, a phenyl or benzyl group;

^B a -CO2R6 group, with R6 representing a hydrogen, a linear or branched alkyl group comprising from 2 to 4 carbon atoms or a benzyl group;

^H a group - R7R ₈ , with R ₇ and R§ representing, independently of each other, a hydrogen, a linear or branched alkyl group of 1 to 4 carbon atoms, a phenyl or benzyl group or with -NR R ₇ taken together represent a saturated 5- or 6-membered ring comprising one or more heteroatoms selected from oxygen and nitrogen, preferably representing a morpholine or piperidine;

^H a carboxamide group -CONRvRg _, with -NR ₇ Rs as defined above;

^B a phenyl group substituted or not by:

one or more halogens selected from fluorine or bromine;

a group-OR ₅ , with R ₅ having the same definition as above; a phenyl or thienyl group;

A 5- or 6-membered aromatic heterocycle comprising 1 or 2 heteroatoms chosen from oxygen, nitrogen and sulfur; or

^{B is} a 5 or 6 membered saturated 5- or 6-membered cyclic or saturated 5- or 6-membered heterocyclic compound having 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur;

a phenyl group substituted or not by:

^B one or more halogens selected from fluorine or bromine; or

^■ a group -OR.5, with R5 having the same definition as above;

^B phenyl;

^B a 5- or 6-membered aromatic heterocycle comprising 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur;

d) R4 represents:

a hydroxyl group;

a group -NR ₇ R ₈ , with R ₇ and R being, independently of each other, a hydrogen, a linear or branched alkyl group of 1 to 4 carbon atoms, a phenyl or benzyl group, or with -NR ₇ Rg taken together representing a 5- or 6-membered heterocycle, having one or more heteroatoms selected from N or O;

an alkoxy group -OR9, with R9 representing:

^B a linear or branched alkyl group having from 2 to 6 carbon atoms;

B a benzyl group;

"-CHR ₁₀ group -COORn, _{-CHR] 0} -OC (= O) R _u, -CHR _OJ or -C (= 0) -ORn wherein Rio and Ru are each independently of one another, a linear or branched alkyl group of 1 to 6 carbon atoms.

The compounds of the present invention may be in the form of pharmacologically acceptable addition salts, such as the addition salts of the compounds of formula (I) with pharmacologically acceptable organic or inorganic bases. Advantageously, the compounds have the general formula (I) in which R represents a hydrogen or a -SC (Ri ₂ ) -CO-CH ₂ -CH (R ₃ ) -CO-R 4 group, R 1, R ₂ , R ₃ and being such that the compound of formula (I) is symmetrical, with R _ls R _2, R ₃ and R 4 can be as defined above or below. Preferably, R is hydrogen.

In particular embodiments of the invention, the compounds have the general formula (I) in which R 1 represents a linear or branched alkyl group of 1 to 6 carbon atoms or a linear or branched alkyl group of 1 to 6 substituted carbon atoms by:

B a phenyl group;

^■ a phenyl group substituted by one or more halogens selected from fluorine or bromine;

^H a 5- or 6-membered saturated cyclic compound or saturated 5- or 6-membered heterocyclic ring containing 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur;

and R ₂ represents a hydrogen, or

R 1 and R ₂ are identical and represent a linear or branched alkyl group of 1 to 6 carbon atoms, or

-C (R _! R ₂ ) - taken together represents:

^B a 5-membered saturated cyclic compound;

^■ a cyclic compound saturated 5-membered ring fused to an aromatic ring;

^H a cyclic saturated compound to 6-membered ring; or

^B a 6 membered saturated heterocyclic compound comprising 1 heteroatom, at the 4-position, selected from oxygen, nitrogen and sulfur;

and R, R ₃ and R 4 are as described above.

More particularly, R 1 may represent an isobutyl group or a methyl group substituted with:

^■ a phenyl group;

^■ a phenyl group substituted in position 4 by halogen selected from fluorine or bromine; a phenyl group substituted in the 4-position by a phenyl group;

and R.2 represents a hydrogen, or

R 1 and R ₂ are identical and represent a methyl or ethyl group, or

-C (RjR ₂ ) - taken together represents:

^■ a cyclic moiety, saturated 5 or 6 membered ring; or

has a 5-membered saturated cyclic group attached to an aromatic ring.

In particular embodiments of the invention, R 1, R 1, R ₂ and R 4 are as described above or below and R ₃ represents a linear or branched alkyl group of 1 to 6 carbon atoms. This alkyl group may be unsubstituted or substituted by a phenyl group. This phenyl group can itself be unsubstituted or substituted by:

one or more halogens selected from fluorine or bromine;

a group-OR5, with R ₅ having the same definition as above;

a phenyl or thienyl group

Advantageously, R ₃ represents a linear or branched alkyl group of 1 to 6 carbon atoms substituted with:

^H a phenyl group; or

a phenyl group substituted with:

one or more halogens taken from fluorine or bromine; a phenyl or thienyl group,

and R, R 1, R ₂ and R ₄ are as described above.

More particularly, R may represent an alkyl group with a carbon substituted with:

a phenyl group;

a phenyl group substituted in the 4-position by a halogen taken from fluorine or bromine;

a phenyl group substituted in the 4-position by a phenyl group.

In particular embodiments of the invention, R, Ri, R ₂ and R? are as described above or below and R ₄ represents:

a hydroxyl group; an alkoxy group -OR9, with R9 representing:

^■ an alkyl, linear or branched, having from 2 to 6 carbon atoms;

B a benzyl group;

and R, R 1, R ₂ and R 3 are as described above.

In particular embodiments of the invention, the compounds have the general formula (I) wherein R is hydroxyl and R, R 1, R 2 and R 3 are as described above.

Preferred compounds have the general formula (I) in which

a) R represents a hydrogen;

b) R] represents a linear or branched alkyl group of 1 to 6 carbon atoms or a linear or branched alkyl group of 1 to 6 carbon atoms substituted with:

^H a phenyl group;

a phenyl group substituted with one or more halogens selected from fluorine or bromine;

and R2 is hydrogen, or

R 1 and R ₂ are identical and represent a linear or branched alkyl group of 1 to 6 carbon atoms; or

-C (R] R2) - taken together represents:

^■ a cyclic compound saturated 5-membered ring;

a 5-membered saturated cyclic compound attached to an aromatic ring;

A 6-membered saturated cyclic compound; or

H is a saturated 6-membered heterocyclic compound comprising 1 heteroatom, at the 4-position, selected from oxygen, nitrogen and sulfur;

c) R3 represents a linear or branched alkyl group of 1 to 6 carbon atoms substituted with: ^■ a phenyl group; or

^■ a phenyl group substituted by:

one or more halogens selected from fluorine or bromine;

a phenyl or thienyl group;

d) R represents a hydroxyl group.

Very particularly preferred compounds have the general formula (I) wherein:

- Rj = CH ₂ Ph; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- Ri = CH ₂ Ph; R ₂ = H; R ₃ = CH ₂ Ph; or

- R 1 = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Ph-Ph); or

- R 1 = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ Ph; or

- R 1 = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

R ₁ = CH ₂ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ Ph; or

- R _! = CH ₂ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- R _! = CH ₂ (4-Ph-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- Ri = CH ₃ ; R ₂ = CH ₃ ; R ₃ = CH ₂ (4-Br-Ph); or

- Ri = C ₂ H ₅ ; R ₂ = C ₂ H ₅ ; R ₃ = CH ₂ (4-Br-Ph); or

C (R R ₂ = Cyclopentyl; R ₃ = CH ₂ (4-Br-Ph); or

- C (R ₁ R ₂ ) - = Cyclohexyl; R ₃ = CH ₂ (4-Br-Ph); or

- C (RiR ₂ ) - = Phenylcyclopentyl (Indanyl); R ₃ = CH ₂ (4-Br-Ph); or

- C (RiR ₂ ) - = Cyclohexyl; R ₃ = CH ₂ (4-Ph-Ph); or

- C (RiR ₂ ) - = Cyclopentyl; R ₃ = CH ₂ (4-Ph-Ph); or

- C (R, R ₂ ) - = Phenylcyclopentyl (Indanyl); R ₃ = CH ₂ (4-Ph-Ph).

Preferred compounds have formula (I) wherein:

- R 1 = CH ₂ CH (CH ₃ ) ₂ ; ₂ = H; R ₃ = CH ₂ (4-Ph-Ph); or

- R 1 = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- C (R, R ₂ ) - = Cyclohexyl; R ₃ = CH ₂ (4-Br-Ph); or

- C (R, R ₂ ) - = Cyclohexyl; R ₃ = CH ₂ (4-Ph-Ph).

The compounds of the present invention may have the general formula (I) wherein R is hydrogen and R4 is hydroxyl. In preferred embodiments, the compounds of the present invention have the general formula (I) wherein R is hydrogen, R ₄ is a hydroxyl group and

- R, = CI LiPh; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- Ri = CH ₂ Ph; R ₂ = H; R ₃ = CH ₂ Ph; or

- R 1 = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Ph-Ph); or

- R 1 = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ Ph; or

- R 1 = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- Ri = C¾ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ Ph; or

- Ri = CH ₂ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- Ri = CH ₂ (4-Ph-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- Ri = CH ₃ ; R ₂ = CH ₃ ; R ₃ = CH ₂ (4-Br-Ph); or

- R = C ₂ H ₅ ; R ₂ = C ₂ H ₅ ; R = CH ₂ (4-Br-Ph); or

- C (RiR ₂ ) - = Cyclopentyl; R ₃ = CH ₂ (4-Br-Ph); or

- C (RiR ₂ ) - = Cyclohexyl; R ₃ = CH ₂ (4-Br-Ph); or

- C (RiR ₂ ) - = Phenylcyclopentyl (Indanyl); R ₃ = CH ₂ (4-Br-Ph); or

- C (RiR ₂ ) - = Cyclohexyl; R ₃ = CH ₂ (4-Ph-Ph); or

- C (RiR ₂ ) - = Cyclopentyl; R ₃ = CH ₂ (4-Ph-Ph); or

- C (RiR ₂ ) - = Phenylcyclopentyl (Indanyl); R ₃ = CH ₂ (4-Ph-Ph).

The compounds of the present invention can be used as a medicine. More particularly, the compounds can be used to prepare pharmaceutical compositions comprising as active ingredient at least one of the compounds described above in combination with at least one pharmaceutically acceptable excipient. Said excipients are selected according to the pharmaceutical form and the desired mode of administration from the usual excipients which are known to those skilled in the art.

Since the compounds of the present invention inhibit the enzymatic activities of neprilysin, these compounds and the compositions containing them can therefore be used in the treatment of pathologies in which the substrates of neprilysin are involved. The present invention also relates to the use of a compound of formula (I) or a composition according to the invention for the manufacture of a medicament for treating a pathology in which the substrates of neprilysin are involved.

For example, by inhibiting the degradation of enkephalins, the compounds of the present invention prove to be effective antidepressants and / or anxiolytics. Advantageously, the compounds of the present invention also appear to possess anti-diarrheal properties by protecting enkephalins from the action of neprilysin. By their mode of action, they can also be effective anti-secretory agents.

The compounds of the present invention may also prove to be effective wound healing agents.

They can be useful to regulate the water balance in the body, such as to reduce cardiac water overload, treat edema, eliminate various fluidic effusions (joints, pleura, ascites), reduce the intraocular pressure and thus treat the glaucoma.

Thus, the compounds of the present invention may be useful for at least one use chosen from the following uses: antisecretory, anxiolytic, antidepressant, cicatrisant, regulation of water balance in the body, reduction of cardiac water overload, treatment of edema or treatment of glaucoma.

The pharmaceutical compositions according to the invention may be administered parenterally, such as intravenously, intrathecally or intradermally, or orally, topically or nasally.

Parenteral dosage forms include aqueous suspensions, isotonic saline solutions or sterile and injectable solutions which may contain pharmacologically compatible dispersing agents and / or wetting agents. Orally administrable forms include tablets, soft or hard capsules, dragees, powders, granules, oral solutions (syrup, oral suspensions). Nasal administrable forms include aerosols, drops. Topically administrable forms include patches, gels, creams, ointments, lotions, sprays, eye drops.

The effective dose of a compound of the invention varies according to many parameters such as, for example, the chosen route of administration, weight, age, sex, the progress of the pathology to treat and the sensitivity of the individual to treat. The present invention, according to another of its aspects, also relates to a method for treating the pathologies indicated above which comprises the administration, to a patient in need thereof, of an effective dose of a compound according to the invention, or a pharmaceutically acceptable salt or a composition according to the invention, preferably parenterally, orally, topically or nasally. The compounds of the present invention can also be used in cosmetology. More particularly, the compounds of the present invention may be employed to prepare cosmetic compositions comprising at least one of the above described compounds. combination with at least one acceptable excipient. These cosmetic compositions can be used to reduce wrinkles and prevent skin aging.

Thus, in another aspect, the present invention relates to a method for reducing wrinkles and preventing skin aging, comprising applying to the skin a cosmetic composition comprising at least one of the compounds described above. combination with at least one acceptable excipient.

The compounds of the present invention can be obtained by various methods.

Thus compounds of formula I for which R = R ₂ = H and R ₄ = OH are obtained in 5 steps from an amino acid II of defined absolute configuration, preferably (R).

Step 1:

Amino acid II is converted to brominated derivative III by a deamination-halogenation reaction, which is generally carried out with configuration retention (Claeson G. et al., (1968) Acta Chem., Scand., 22, 3155-3159; A. et al (1987) J. Chem Soc Perkin Trans.1, 111-120).

The brominated derivative III is converted into thioether IV by nucleophilic substitution with inversion of configuration by the action of 4-methoxy-a-toluenethiol (PMBSH) in basic medium.

Step 2: Preparation of halomethyl cctone VI from IV

Process 1: compound IV is converted into ketene V either from the mixed anhydride of IV (prepared by the action of isobutylchloroformate and N-methylmorpholine), or from the acid chloride (prepared by the action of chloride thionyl on IV).

IV v The ketene V is then converted to halomethylketone VI by bubbling with HCl gas or HBr gas in -Dioxane, diethylether or ethyl acetate.

Process 2: alternatively chloromethylketone VI (X = Cl) can be obtained by the action of chloroiodomethane on the methyl ester of IV (prepared in the presence of DMAP and EDCI or else by the action of acetyl chloride in methanol) in the presence of freshly prepared LDA (Chen et al (1997) Tet, Lett 38, 18, 3175-3178).

Step 3

Process 1: Nal-treated halogenomethylketone VI undergoes halogen exchange and then reacts with the sodium salt of dialkylmalonate to give compound VII. R ₂ may be a methyl, ethyl or tert-butyl group.

VI

VII

The substituent R ₃ is introduced by the action of a brominated derivative R Br on the anion of the preceding malonate VII, deprotonated in situ by NaH. Compound VIII is obtained.

VII VIII

Process 2: the substituent R ₃ may also be introduced directly on the halogenomethylketone VI, in the case where X = Br. The latter, treated with NaI, undergoes halogen exchange and then reacts with the sodium salt of the substituted dialkylmalonate to give compound VIII.

This reaction makes it possible to preserve the configuration of the carbon carrying the thiol.

Step 4

After hydrolysis of the VIII esters by the action of TFA (where R ₂ is a tert-Butyl group) or by saponification (where R ₂ is a methyl or ethyl group), a reflux decarboxylation in toluene, for example, leads to compound IX.

Step 5

The deprotection of the thiol present on compound IX is carried out in 2 steps by the action of DTNP (2,2'-dithiobis (5-nitropyridine)) in trifluoroacetic acid followed by a reaction with TCEP (tris (2- carboxyethyl) phosphine) (Harris KM et al (2007) J Pept Sci (2), 81-93) or directly by heating in trifluoroacetic acid at 50 ° C in the presence of anisole to give the mercaptoalkanoic acid I.

PMBS

Alternatively, for the compounds I in which R 1 = R ₂ = alkyl, R = H and R = OH, the chloromethylketones VI can be prepared from a reaction between bromoacetic acid and 4-methoxybenzyl mercaptan followed by double alkylation and conversion to chloromethylketone VI as previously described.

In the case where Ri and R form a cycle.

Process 1: Chloromethyl ketone VI is prepared directly from the corresponding ester (for example methyl) by alkylation using 4-methoxy benzyl mercaptan disulfide (or from another activation of this thiol) and transformation in chloromethylketone VI.

Method 2: alternatively, the cyclic glycerine-methyl ketone VI can be prepared directly from the corresponding ester (methyl for example) by alkylation with activated 4-methoxy benzyl mercaptan and conversion to bromomethyl ketone VI using a TMSN solution ₂ .

The rest of the synthesis is carried out as described previously.

In the case where R ₃ is 4-Bromobenzyl, compound IX can undergo a Suzuki reaction to introduce

Compounds IX, when R ₂ = H, have 2 centers of asymmetry and consist of 4 stereoisomers. When R 1 = R ₂ = alkyl or ring, the IX compounds have a single center of asymmetry and are therefore a mixture of 2 stereoisomers.

The compounds of the present invention encompass all stereoisomers of I, obtained after deprotection of IX.

In the case where the IX compounds are chiral, they can be separated by selective precipitation with chiral amines such as Γ-methylbenzyl amine or norephedrine or chiral column HPLC.

EXAMPLES

The invention will be further illustrated without being limited by the following examples. Step 1: synthesis of (R) -2-Bromo carboxylic acids

The amino acid of configuration (R) - (39.3 mmol) is solubilized in 50 ml of water. At 0 ° C., KBr (3.5 eq, 31.8 g) and then H 2 SO 4 (7.73 ml) are added dropwise, while checking that the temperature remains below 5 ° C. The mixture is cooled to -10 ° C and NaN0 ₂ (1.3 eq, 3.59 g) solubilized in 17 mL of water is added dropwise. The mixture is stirred for 2 hours at -5 ° C.

After returning to ambient temperature, the mixture is extracted with CH ₂ Cl ₂ (2 * 50mL). The organic phase is washed with H ₂ O, sat. NaCl, dried over Na ₂ SO ₄ to give the expected product of configuration (R).

III a R 1 = CH ₂ Ph: light yellow oil; (Yield: 50%); Rf (CH ₂ Cl ₂ / MeOH) 0.62

NMR (CDCl ₃ , 200 MHz): 3.15-3.40 (2H, dd); 4.69 (1H, m); 7.20-7.40 (5H, m)

IIIb R = CH ₂ CH (CH ₃ ) ₂ : oil; (Yield: 82.5%); Rf (CH ₂ Cl ₂ / MeOH): 0.49

NMR (CDCl ₃ , 200 MHz): 0.90-1 .0 (6H, m); .55 (2H, m); 2.40 (1H, m); 4.29 (1H, d) III c R 1 = CH ₂ (4-Br-Ph): light yellow oil; (Yield: 50%); Rf (CH ₂ Cl ₂ / MeOH) 0.62

NMR (CDCl ₃ , 200 MHz): 3.15-3.40 (2H, dd); 4.70 (1H, m); 7.20-7.40 (4H, m)

IIId R] = CH ₂ (4-Ph-Ph): light yellow oil; (Yield: 60%); Rf (CH ₂ Cl ₂ / MeOH): 0.7

NMR (CDCl ₃ , 200 MHz): 3.15-3.40 (2H, dd); 4.70 (1H, m); 7.20-7.60 (9H, m)

Synthesis of aci (4-methoxybenzylthio) carboxylic acids

Under an inert atmosphere, 4-methoxybenzyl mercaptan (4.2 mL, 30.06 mmol, 1 eq) is solubilized in 70 mL of anhydrous THF and 1 eq of 60% NaH (1.33 g, 33.07 mmol) are added. The mixture is stirred for 15 min at room temperature and the brominated derivative 3 (1 eq, 30.06 mmol) solubilized in 30 mL of THF is added dropwise using a dropping funnel. The mixture is stirred overnight at room temperature. The mixture is evaporated to dryness and is taken up in AcOEt. The organic phase is washed with H ₂ O, sat. NaCl, dried with Na ₂ S0 ₄ , evaporated under reduced pressure to give the crude product. This is purified by chromatography on silica with CHex / AcOEt 5/5 as elution system to give compound IV of configuration (S) as an oil.

IV a R 1 = CH ₂ Ph: oil; (Yield: 40%); Rf (CH ₂ Cl ₂ / MeOH / 9/1): 0.5

NMR (CDC1 _3, 200 MHz): 2.80-3.10 (2H, m); 3.68 (2H, s); 3.76 (3H, s); 4.53 (1H, t); 6.84 (2H, d); 7.09-7.49 (7H, m)

IV b = CH ₂ CH (CH ₃ ) ₂ : oil; (Yield: 26%); Rf (CHex / AcOEt): 0.65

NMR (CDCl ₃ , 200 MHz): 0.90-1.0 (6H, m); 1.55 (2H, m); 2.25 (1H, m); 3.40 (1H, d); 3.70 (2H, s); 3.90 (3H, s); 6.8-6.9 (4H, m)

IV c R 1 = CH ₂ (4-Br-Ph): oil; (Yield: 40%); Rf (CH ₂ Cl ₂ / MeOH / 9/1): 0.5

NMR (CDCl3, 200 MHz): 3.0-3.30 (2H, m); 3.60 (1H, q); 3.70 (2H, s); 3.90 (3H, s); 6.80- 7.20 (8H, m)

IVdRi = CH ₂ (4-Ph-Ph): oil; (Yield: 50%); Rf (CH ₂ Cl ₂ / MeOH / 9/1): 0.6

NMR (CDCl ₃ , 200 MHz): 3.0-3.30 (2H, m); 3.60 (1H, q); 3.70 (2H, s); 3.90 (3H, s); 6.80-7.2 (13H, m)

2nd step:

Method 1:

Method 1: Synthesis of diazoketone from mixed anhydride

To a solution of IV acid (18.5 mmol) in 20 mL of dry THF under an inert atmosphere, N-methyl morpholine (2.15 mL, 1.05 eq) and iBuOCOCl (2.52 mL, 1.05) are successively added at -20 ° C. eq). The mixture is stirred for 5-10 min at -20 ° C and then the precipitate is filtered through Celite and washed with 20 mL THF.

CH ₂ N ₂ solution in ether (2.5 eq) (previously prepared from Diazald ^® and KOH in carbitol), is transferred to the solution of activated ester at 0 ° C. The solution becomes yellow. The mixture is stirred for 2 hours at room temperature.

Method 2: Synthesis of the diazoketone from the acid chloride

IV V The IV acid (14.5 mmol) is solubilized in 23 mL of anhydrous CH ₂ C1 ₂ . SOCl ₂ (1.5 eq; 21.75 mmol) is added at ambient temperature and the mixture is heated under reflux for 2 hours under an inert atmosphere. The mixture is then evaporated to dryness to give a brownish oil. The product is solubilized in anhydrous THF at a rate of 5 mmol / mL.

The previously prepared solution of CH ₂ N ₂ (2.5 eq) in ether is transferred to the acid chloride solution at 0 ° C. The solution becomes yellow. The mixture is stirred for 2 hours at room temperature under an inert atmosphere.

Synthesis of Chloromethyl Ketone

The solution of the compound V is placed, under an inert atmosphere, in a tricolor maintained at 0 ° C. The mixture is saturated with HCl, bubbling at 0 ° C.

After 30 min, the solvent and the excess HCl are evaporated under reduced pressure. The product is taken up in AcOEt (150 mL) and is then washed with NaHCO 3> 10%, H ₂ O and dried over Na ₂ SO ₄ to give the product in the crude state. The latter is used as it is without purification for the next step.

Process 2: Synthesis of Methyl Ester

IV VI

Acetyl chloride (3eq; 2mL) is added dropwise at 0 ° C., under an inert gas, to a solution of acid IV (9.1 mmol) in 50 ml of anhydrous MeOH. The mixture is stirred overnight at room temperature. The mixture is concentrated under reduced pressure, taken up in MTBE (mhthyl-tert-butyl ether) (200 ml). The organic phase is washed with NaHCO 3> 10% (100 ml), H ₂ 0 (100 ml) and sat. NaCl. (1 OOmL). The organic phase is dried over Na ₂ SO ₄ and is then concentrated under reduced pressure to give the crude methyl ester. The latter is chromatographed by flash system on silica gel.

A solution of LDA (5 eq) in THF (55 mL), freshly prepared from 1.6M BuLi in hexane (15 mL) and diisopropylamine (3.4 mL), was added dropwise at room temperature. min to methyl ester (4.42 mmol) and chloroiodomethane (1, 3 mL, 4 eq) in solution in 25 mL of THF. The internal temperature of the reaction is kept below -70 ° C during the addition and at -75 ° C for 10 min. A solution of acetic acid (6 mL in 44 mL of THF) is added keeping the temperature below -65 ° C to neutralize the medium. The mixture is then extracted with AcOEt. The organic phase is washed with 10% NaHCO _3, 10% citric acid, saturated NaCl, dried over Na ₂ SO ₄ and then concentrated under reduced pressure to give the crude product which is used as it is for the next step.

VI to R = CH ₂ Ph; R ₂ = H: orange oil; (Yield: 93.0%); Rf (CHex / AcOEt 6/4): 0.73 HPLC: romasil C18 CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) 60/40 Rt: 19.18 min

NMR (CDC1 _3, 200MHz): 2.85 (1H, dd); 3.2 (1H, dd); 3.55 (1H, d); 3.6 (2H, d); 3.7 (3H, s); 4.1 (2H, d); 6.7 (2H, d); 7.2-7.4 (7H, m)

VI b R = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H: orange oil; (Yield: 94.0%); Rf (CHex / AcOEt 5/5): 0.68 NMR (CDCl ₃ , 200MHz): 0.90-1.0 (6H, m); 1.55 (2H, m); 2.25 (1H, m); 3.40 (1H, d); 3.70 (2H, s); 3.90 (3H, s); 4.25 (2H, d); 6.80 (2H, d); 7.15 (2H, d)

VI c R 1 = CH ₂ (4-Br-Ph); R ₂ = H: orange oil; (Yield: 85.0%); Rf (CHex / AcOEt 6/4): 0.80 NMR (CDCl ₃ , 200MHz): 2.85 (1H, dd); 3.2 (1H, dd); 3.55 (1H, d); 3.6 (2H, d); 3.7 (3H, s); 4.1 (2H, s); 6.7 (2H, d); 7.2-7.4 (6H, m)

VI d R 1 = CH ₂ (4-Ph-Ph); R ₂ = H: orange oil; (Yield: 90.0%); Rf (CHex / AcOEt 6/4): 0.73 NMR (CDCl ₃ , 200MHz): 2.85 (1H, dd); 3.2 (1H, dd); 3.55 (1H, d); 3.6 (2H, d); 3.7 (3H, s); 4.1 (2H, s); 6.7 (2H, d); 7.2-7.5 (11H, m)

Synthesis of geminal chloromethylketones

Bromoacetic acid (10 g, 72 mmol) is dissolved in an inert atmosphere in 50 ml of MeOFI. 11 mL (1.1 eq) of 4-methoxybenzyl mercaptan are added at 4 ° C. and an alcoholic sodium hydroxide solution (6.4 g of NaOH (2.2 eq) in solution in 100 mL of MeOH) is added dropwise. The mixture is stirred for 40 min at room temperature. The solvent is evaporated under reduced pressure. The product is taken up in Et ₂ O (200 mL) and 350 mL of NaHCO ₃ 10%. The aqueous phase is acidified to pH = l and then extracted with 350 mL of Et ₂ 0. The organic phase is washed with H ₂ 0 (lOOmL), NaCl sat (lOOmL) and dried over Na ₂ S0 ₄ to give 15 g of a crude white solid (Yield: 98%) which is used as it is for the next step.

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 10-90% 15 min, Rt = 10.64 min

NMR (CDC1 _3, 200MHz): 3.0 (2H, s); 3.75 (3H, s); 3.75 (2H, s); 6.80 (2H, d); 7.20 (2H, d) Acetyl chloride (1.5eq, 7.6mL, 106 mmol) is added dropwise at 4 ° C, under an inert atmosphere, to a solution of the above acid (70.8 mmol) in 150 mL anhydrous MeOH. The mixture is stirred overnight at room temperature. The mixture is concentrated under reduced pressure, taken up in MTBE (methyl tert-butyl ether) (350 ml). The organic phase is washed with 0.5N HCl (2 * 100mL), 10% NaHCO ₃ (2 * 100mL), H ₂ O (100mL) and sat. NaCl. (LOOmL). The organic phase is dried over Na ₂ SO ₄ and is then concentrated under reduced pressure to give the methyl ester.

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 50-90% 10 min, Rt = 5.24min

NMR (CDCl3, 200MHz): 3.1 (2H, s); 3.75 (3H, s); 3.83 (2H, s); 3.85 (3H, s); 6.85 (2H, d); 7.30 (2H, d).

A solution of 1M LiHMDS THF (4.4mL; leq) is added dropwise, under an inert atmosphere, at -78 ° C., to a solution of the above ester (1 g; 4.4 mmol; leq) solubilized in 5 mL of anhydrous THF. . The mixture is stirred at -78 ° C. for 1 h and then the solution of the RX derivative (1 eq) is added under an inert atmosphere at -78 ° C. The mixture, brought to room temperature, is stirred for 3 hours. The mixture is again cooled to -78 ° C and 1 LiHMDS is added followed by 1 eq of RX. The mixture, brought to ambient temperature, is stirred for 4 hours. The mixture is then partitioned between 200 mL IN HCl and 300 mL AcOEt. The organic phase is dried over Na ₂ SO ₄ and is then concentrated under reduced pressure to give the crude product which is purified by chromatography on silica gel.

R 1 = CH ₃ , R ₂ = CH ₃ : oil (Yield: 44%)

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 50-90% 10 min, Rt = 7.39 min

NMR (CDCl ₃ , 200MHz): 1.60 (6H, s); 3.70 (3H, s); 3.80 (2H, s); 3.80 (3H, s); 6.85 (2H, d); 7.25 (2H, d)

Ri = C ₂ ¾, R ₂ = C ₂ H ₅ : oil (Yield: 55%) HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 50-90% 10 min, Rt = 9.72 min

NMR (CDCl3, 200MHz): 0.90 (6H, t); 1.85 (4H, m); 3.68 (2H, s); 3.70 (3H, s); 3.80 (3H, s); 6.82 (2H, d); 7.22 (2H, d)

Chloromethylketone is synthesized as described in 4b

Vi e R ₁ = CH ₃ , R ₂ = CH ₃ : amber oil

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 50-90% 10 min, Rt = 7.91 min

NMR (CDCl ₃ , 200MHz): 1.60 (6H, s); 3.55 (1H, d); 3.60 (2H, s); 3.80 (3H, s); 4.45 (2H, s); 6.7 (2H, d); 7.2 (2H, d)

VIf R, = C ₂ H ₅ , R ₂ = C ₂ H ₃ : amber oil

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 50-90% 10 min, Rt = 10.04 min

NMR (CDCl3, 200MHz): 0.85 (6H, t); 1.80 (4H, q); 3.55 (1H, d); 3.60 (2H, s); 3.80 (3H, s); 4.45 (2H, s); 6.7 (2H, d); 7.2 (2H, d)

Synthesis of cyclic chloromethylketones

Method 1:

To a solution in THF (9 mL), under an inert atmosphere, DIPA (diisopropylethylamine) (3 mmol, 1.3 eq, 420 μL) is introduced at -10 ° C., the 2.5M solution of BuLi in hexane (2.77 mmol; lmL, 1.2eq). The mixture is stirred at 0 ° C. This LDA solution, freshly prepared, is added dropwise to a solution of cyclopentanoic acid methyl ester in 5 mL of THF at -55 ° C. The mixture is stirred at -55 ° C. under an inert atmosphere. HMPA (hexylmethylphosphoramide) (3.46 mmol, 1.5eq, 610μί) is added and the mixture is stirred for 10 min at the same temperature. A solution of 4-methoxybenzyl mercaptan disulfide (3 mmol, 1.3 eq, 920 mg) in 12 mL of THF is then added dropwise at -55 ° C. After returning to ambient temperature, the mixture is stirred overnight. The mixture is partitioned between 10mL NH ₄ CI sat. and 20mL AcOEt. The organic phase is washed with NH ₄ C1 sat. (2 * 10 mL), NaCl sat (2 * 15 mL), dried over Na ₂ SC> 4 and then concentrated under reduced pressure to give the crude product which is purified by chromatography on silica gel.

C (R] R ₂ ) = Cyclopentyl: oil (Yield: 40%)

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 30-90% 10 min, Rt = 9.68 min

NMR (CDCl ₃ , 200MHz): 1.60-2.40 (8H, m); 3.67 (3H, s); 3.77 (2H, s); 3.80 (3H, s); 6.83 (2H, d); 7.24 (2H, d)

Chloromethylketone is synthesized as described in 4b

VI g C (Ri R ₂ ) = Cyclopentyl: amber oil

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 60-90% 10 min, Rt = 6.47 min

NMR (CDCl ₃ , 200MHz): 1.60-1.80 (8H, m); 3.54 (2H, s); 3.80 (3H, s); 4.47 (2H, s); 6.78 (2H, d); 7.22 (2H, d)

VI h C (RiR ₂ ) = Cyclohexyl: amber oil

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 70-90% 10 min, Rt = 8.11 min

NMR (CDCl ₃ , 200MHz): 1.40-2.20 (10H, m); 3.45 (2H, s); 3.80 (3H, s); 4.40 (2H, s); 6.80 (2H, d); 7.20 (2H, d)

Method 2

To a solution in THF (10 ml), under an inert atmosphere, DIPA (diisopropylethylamine) (16.7 mmol, 1.2 eq, 2.34 ml) is introduced at -10 ° C., the 2.5M solution BuLi in hexane (16 mmol, 6.4mL, 1.15eq). The mixture is stirred at 0 ° C. This freshly prepared solution of LDA is added dropwise to a solution of phenylcyclopentanoic acid methyl ester in 5 mL of THF and HMPA (hexylmethylphosphoramide) (1.Oeq, 2.5 mL) at -78 ° C. The mixture is stirred at -78 ° C. under an inert atmosphere. The activated 4-methoxybenzyl mercaptan thiol (18 mmol, 1.3 eq, 6.37 mg) is added in the solid state at -78 ° C. The mixture is stirred for 1 hour at -78 ° C. The mixture is partitioned between 200 mL IN HCl and 200 mL AcOEt. The organic phase is diluted with 200 mL AcOEt, washed with sat. NaCl (200 mL), dried over Na ₂ SC 4 and then concentrated under reduced pressure to give the crude product which is purified by chromatography on silica gel.

C (R, R ₂ ) = PhenylCyclopentyl: white solid (Yield: 25%)

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 50-90% 10 min, Rt = 9.59 min

NMR (CDC1 _3, 200MHz): 2.90-3.70 (6H, m); 3.55 (3H, s); 3.65 (3H, s); 6.60-7.10 (8H, m) The product of the previous step (1.13 g, 3.44 mmol) is solubilized in 14 mL of a THF / MeOH mixture. 14 ml of 2N NaOH are added and the mixture is stirred for 3 hours at room temperature. The mixture is diluted with 40 mL H ₂ O. The mixture is concentrated under reduced pressure. The aqueous phase is acidified with 1N HCl and then extracted with MTBE (3 × 100 mL), washed with sat. NaCl (100 mL), dried over Na ₂ SO ₄ and then concentrated under reduced pressure to give a white solid (Yield: 98% ).

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H _z O (0.1% TFA) Gradient 50-90% 10 min, Rt = 6.50 min

The oxalyl chloride (2 mmol, 1.5eq, 177 μΚ) is added dropwise, under an inert atmosphere, at 0 ° C., to a solution of the above acid (1.38 mmol, 434 mg) in 10 ml CH ₂ C1 ₂ , in the presence of 20μΕ DMF (0.2 eq). The mixture is allowed to come to room temperature and is then stirred for 30 minutes at room temperature.

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 70-90% 10 min, Rt = 6.70 min.

After evaporation under reduced pressure, the acid chloride is taken up, under an inert atmosphere, in anhydrous CH ₃ CN (5 mL) and TMSCHN ₂ (1M in Et ₂ O) (1.5 eq, 1 mL) is added dropwise at 0 °. vs. The mixture is allowed to come to room temperature and is then stirred for one hour. The mixture is then trapped with 300 μί (1.65 mmol, 1.2 eq) HBr 33% in acetic acid. The mixture is stirred for 15 minutes at room temperature. The mixture is concentrated under reduced pressure and is taken up in MTBE (200 mL). The organic phase is washed with NaHC (10% (100 mL), sat. NaCl (50 mL), dried over Na SO 4 and then concentrated under reduced pressure to give compound 6h as a brown oil (yield: 83%). (RiR ₂ ) = Phenylcyclopentyl (Indanyl): white solid

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 70-90% 10 min, Rt = 5.30 min

NMR (CDCl ₃ , 200MHz): 2.90-3.70 (6H, m); 3.65 (3H, s); 4.15 (2H, s); 6.60-7.10 (8H, m)

Step 3

Method 1

Under an inert atmosphere, 980 mg of 60% NaH (1 eq) is added to the dialkylmalonate (1 eq) in 25mL (1mL / mmol) of DME (1,2-dimethoxyethane). The mixture is stirred at room temperature.

A mixture of chloromethylketone VI (24.42 mmol) and NaI (24.42 mmol, 3.66 g, 1 eq) in 50 mL of DME is stirred at room temperature for 15 minutes and then added to a freshly prepared solution of dialkylmalonate sodium salt. The mixture is stirred for 4 hours at room temperature.

At the end of the reaction, the solvent is evaporated under reduced pressure. The product is taken up in dichloromethane. The organic phase is washed with water and dried over Na ₂ SC 4. The product is chromatographed on a silica column with CHex / AcOEt 9/1 as the elution system. VII al Ri = CH ₂ Ph, R ₂ = H, R ₁₂ = CH ₂ CH ₃ : orange oil; (Yield: 60%); Rf

(CHex / AcOEt 9/1): 0.16

HPLC: Kromasil Cl 8 CH ₃ CN / H ₂ O (0.1% TFA) 80/20 Rt: 6.57 min

NMR (CDCl ₃ , 200MHz): 1.30 (6H, t); 2.70-3.40 (4H, m); 3.45 (1H, t); 3.50 (2H, d); 3.60 (1H, t); 3.65 (3H, s); 4.15 (4H, q); 6.75 (2H, d); 7.2-7.4 (7H, m)

VII a 2 R 1 = CH ₂ Ph, R ₂ = H, R _n = tBu: orange oil; (Yield: 62%); Rf (CHex / AcOEt

9/1): 0.36

HPLC: Kromasil C18 CH ₃ CN / H ₂ O (0.1% TFA) 85/15 Rt: 15.51 min

NMR (CDCl ₃ , 200MHz): 1.40 (18H, s); 2.70-3.40 (4H, m); 3.45 (1H, t); 3.50 (2H, d); 3.60 (1H, t); 3.65 (3H, s); 6.75 (2H, d); 7.2-7.4 (7H, m) VII b R 1 = CH ₂ CH (CH ₃ ) ₂ , R ₂ = H, R ₁₂ = tBu: orange oil; (Yield: 30%); Rf (CHex / AcOEt 9/1): 0.49

HPLC: Kromasil C18 CH ₃ CN / H ₂ O (0.1% TFA) 90/10 Rt: 6.49 min

NMR (CDCl3, 200MHz): 0.90-1.0 (6H, m), 1.40 (18H, s); 1.55 (2H, m); 2.15 (1H, m); 3.19 (2H, m); 3.40 (2H, m); 3.50 (2H, d); 3.80 (3H, s); 6.75 (2H, d); 7.2 (2H, d)

VII c R 1 = CH ₂ (4-Br-Ph), R ₂ = H, K _n = tBu: orange oil; (Yield: 62%); Rf (CHex / AcOEt 9/1): 0.36

HPLC: Kromasil C18 CH ₃ CN / H ₂ O (0.1% TFA) 85/15 Rt: 15.51 min

NMR (CDCl3, 200MHz): 1.40 (18H, s); 2.70-3.40 (4H, m); 3.45 (1H, t); 3.50 (2H, d); 3.60 (1H, t); 3.65 (3H, s); 6.75 (2H, d); 7.2-7.4 (6H, m)

VIId R] = CH ₂ (4-Ph-Ph), R ₂ = H, R ₁₂ = tBu: orange oil; (Yield: 60%); Rf (CHex / AcOEt 9/1): 0.36

HPLC: Kromasil C18 CH ₃ CN / H ₂ O (0.1% TFA) 85/15 Rt: 16.71 min

NMR (CDCU, 200MHz): 1.40 (18H, s); 2.70-3.40 (4H, m); 3.45 (1H, t); 3.50 (2H, d); 3.60 (1H, t); 3.65 (3H, s); 6.75 (2H, d); 7.2-7.5 (11H, m)

VII e Rj = CH ₃ , R ₂ = CH ₃ , R, ₂ = and (yield: 40%)

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 70-90% 10 min, Rt = 4.87 min

NMR (CDCl3, 200MHz): 1.30 (6H, t); 1.55 (6H, s); 3.40 (2H, d); 3.55 (2H, s); 3.75 (1H, t); 3.82 (3H, s); 4.25 (4H, q); 6.85 (2H, d); 7.25 (2H, d)

VII f R = C ₂ H ₅ , R ₂ = C ₂ H ₅ , R 12 = Et (yield: 30%)

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 50-90% 10 min then 90-50%, Rt = 1 1.56 min

NMR (CDCl ₃ , 200MHz): 0.80 (6H, t); 1.20 (6H, t); 1.75 (4H, m); 3.30 (2H, s); 3.38 (2H, d); 3.65 (1H, t); 3.72 (3H, s); 4.15 (4H, q); 6.72 (2H, d); 7.15 (2H, d)

VII g C (R, R ₂ ) = Cyclopentyl, R, ₂ : and (yield: 24%)

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 60-90% 10 min, Rt = 8.04 min

NMR (CDCl ₃ , 200MHz): 1.29 (6H, t); 1.60-2.30 (8H, m); 3.39 (2H, d); 3.52 (2H, s); 3.74 (1H, t); 3.79 (3H, s); 4.22 (4H, q); 6.83 (2H, d); 7.20 (2H, d)

VII h C (RiR ₂ ) = Cyclohexyl, R, ₂ : and (Yield: 33%)

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 70-90% 10 min, Rt = 7.76 min NMR (CDCl ₃ , 200MHz): 1.29 (6H, t); 1.60-2.30 (10H, m); 3.45 (2H, d); 3.50 (2H, s); 3.75 (1H, t); 3.80 (3H, s); 4.25 (4H, q); 6.85 (2H, d); 7.20 (2H, d)

VII i C (R, R ₂ ) = Phenylcyclopentyl (Inda, ½: And (Yield: 80%)

HPLC: Atlantis T3, CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 70-90% 10 min, Rt = 6. min

NMR (CDCl ₃ , 200MHz): 1.20 (6H, t); 2.90-3.60 (9H, m); 3.65 (3H, s); 4.15 (4H, q); 6.60-7.10 (8H, m)

Alkylation of malonate

To a solution of the product VII in 15 ml of DME (dimethoxyethane) are added 1.5 eq of 60% NaH.

The mixture is stirred at room temperature for 1 h and then the brominated derivative R ₃ Br (3 eq) is added. The mixture is stirred overnight at room temperature.

The solvent is evaporated under reduced pressure and the mixture is taken up in H ₂ O and AcOEt. The organic phase is washed with H ₂ 0, dried over Na ₂ SO ₄ and then concentrated under reduced pressure. The product is chromatographed on silica with CHex / AcOEt 9/1 as elution system to give the desired product VIII.

VIII to R = CH ₂ Ph; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); R ₁₂ = And: orange oil

VIII b R = CH ₂ Ph; R ₂ = H; R ₃ = = CH ₂ Ph; Rj ₂ = tBu: orange oil

VI II c R = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = = H; R ₃ = CH ₂ (4-Ph-Ph); R ₁₂ = And: orange oil

VIIIdRi = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = = H; R ₃ = CH ₂ Ph; R ₁₂ = And: orange oil

VIII e R | = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = = H; R ₃ = CH ₂ (4-Br-Ph); R ₁₂ = And: oran oil, aged

VIII R = CH ₂ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ Ph; R ₁₂ = tBu: orange oil

VIII g Rj = CH ₂ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); Ri ₂ = tBu: orange oil;

VIII h R = CH ₂ (4-Ph-Ph); R ₂ - ^: H; R ₃ = CH ₂ (4-Br-Ph); R, ₂ = tBu: orange oil

VI II i R = CH ₃ ; R ₂ = CH ₃ ; R ₃ = = CH ₂ (4-Br-Ph); R ₁₂ = And: orange oil

VIII R = C ₂ H ₅ ; R ₂ = C ₂ H ₅ ; R 3 = CH ₂ (4-Br-Ph); Ri ₂ = And: orange oil

VI II k C (R, R ₂ ) - Cyclopentyl; R ₃ = CH ₂ (4-Br-Ph); R _{] 2} = And: orange oil

VIII 1 C (R R ₂ ) = Cyclohexyl; R ₃ = CH ₂ (4-Br-Ph); R, ₂ = And: orange oil

VHI m C (R] R ₂ ) = Phenylcyclopentyl (Indanyl); R ₃ = CH ₂ (4-Br-Ph); R, ₂ = And: orange oil Step 4

VIII IX

When R ₁₂ is a methyl or ethyl group, compound VIII (0.585 mmol) is dissolved in 10 mL of EtOH and 2N NaOH (6 eq) is added. The mixture is stirred overnight at room temperature. Ethanol is evaporated under reduced pressure. The product is taken up in water and extracted with Et ₂ O. The aqueous phase is acidified with 3N HCl and is extracted with Et ₂ O. The organic phase is dried over Na ₂ SC 4 and is then evaporated under reduced pressure to give give a yellowish oil.

When R ₁₂ is a tert-butyl group, product VIII is dissolved in 10 ml of CH ₂ C1 ₂ and 10 ml of TFA are added. The mixture is stirred for 2 hours at room temperature. The solvents are evaporated under reduced pressure. The product is taken up in water and extracted with Et ₂ O. The organic phase is dried over Na ₂ SO ₄ and then evaporated under reduced pressure to give a yellowish oil.

The product formed is then solubilized in 6 ml of toluene and is heated at 150 ° C. for 12 hours.

The solvent is evaporated under reduced pressure to give compound IX.

IX to Rj = CH ₂ Ph; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); orange oil

IX b R 1 = CH ₂ Ph; R ₂ = H; R ₃ = CH ₂ Ph; orange oil

IX c R ₍ = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Ph-Ph); orange oil

IX d Rj = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ Ph; orange oil

IX and R 1 = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); orange oil

IX f R = CH ₂ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ Ph; orange oil

IX g R1 = C¾ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); orange oil

IX h R 1 = CH ₂ (4-Ph-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); orange oil

IX i R 1 = CH ₃ ; R ₂ = CH ₃ ; R ₃ = CH ₂ (4-Br-Ph); orange oil

IX = C ₂ H ₅ ; R ₂ = C ₂ H ₅ ; R ₃ = CH ₂ (4-Br-Ph); orange oil

IXk C (RiR ₂ ) = Cyclopentyl; R ₃ = CH ₂ (4-Br-Ph); orange oil

IX 1 C (R ₁ R ₂ ) = Cyclohexyl; R ₃ = CH ₂ (4-Br-Ph); orange oil

IX m C (R ₁ R ₂ ) = Phenylcyclopentyl (Indanyl); R ₃ = CH ₂ (4-Br-Ph); orange oil Suzuki's action

Compound IX 1 (180 mg, 0.356 mmol) is dissolved in 2 ml of toluene under an inert atmosphere. Pd (PPh ₃ ) ₄ (1mg, 3% mol) is added and the mixture is stirred for 5 min at room temperature. Phenylboronic acid (46 mg, 0.374 mmol, 1.05 eq) is added in 1 mL MeOH followed by 500 μL · of 2M Na ₂ CO ₃ . The mixture is refluxed 1:30 and the mixture is concentrated under reduced pressure. The reaction mixture is taken up in 3 μL Et ₂ O and 30 mL 1N HCl. The organic phase is washed with 20 ml 1N HCl, 10 ml sat. NaCl, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The product is chromatographed on silica with Hept / AcOEt 65/35 as the elution system to give the desired product IX f.

IX ii C (R] R ₂ ) = Cyclohexyl; R ₃ = CH ₂ (4-Ph-Ph); 75 mg orange oil (Yield: 47%)

ESI (+): [M + Na] ⁺ = 525

HPLC: Atlantis T3 CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 70-90% 10 min, Rt = 8.60 min

IX o C (RiR ₂ ) = Cyclopentyl; R ₃ = CH ₂ (4-Ph-Ph); orange oil (yield: 25%)

ESI (-): [MH] ^" = 487

HPLC: Atlantis T3 CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 70-90% 10 min, Rt = 6.60 min

IX p C (RiR ₂ ) = Phenylcyclopentyl (Indanyl); R ₃ = CH ₂ (4-Ph-Ph); orange oil (Yield: 55%) ESI (-): [MH] ^" = 535

HPLC: Atlantis T3 CH ₃ CN (0.1% TFA) / H ₂ O (0.1% TFA) Gradient 70-90% 10 min, Rt = 7.45 min

Step 5

## STR2 ## 1 BTFA (1 M), TFA

HS.

_R R ₂ ^R or TFA, Anisole, 50 ° CR 2 R ₂

or 1) DTNP, TFA ¹

i ^X 2) TCEP, CH ₃ CN I

Method 1: Compound IX (0.53 mmol) from step 7 is solubilized in 2.1mL of BTFA (boron tristrifluoroacetate) (1M) (prepared beforehand from BBr ₃ and TFA) and is stirred lh at room temperature. The solvents are evaporated under reduced pressure and the mixture is purified by semi-preparative HPLC.

R = CH ₂ Ph; R ₂ = H; R ₃ = CH ₂ Ph

ESI (+): [M + H] ⁺ = 329

HPLC: ACE Cl 8 CH ₃ CN / H ₂ O (0.1% TFA) Gradient 60/90 in 30 min; Rt: 7.50 min

I b Ri = iBu; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph)

ESI (+): [M + H] ⁺ = 372-374

HPLC: ACE Cl 8 CH ₃ CN / H ₂ O (0.1% TFA) Gradient 60/90 in 30 min; Rt: 8.60 min

Ri = CH ₂ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph)

ESI (+): [M + H] ⁺ = 384

HPLC: ACE C18 CH ₃ CN / H ₂ O (0.1% TFA) Gradient 60/90 in 30 min; Rt: 10.52 min

R 1 = CH ₂ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ Ph

ESI (+): [M + H] ⁺ = 406-408

HPLC: ACE C18 CH ₃ CN / H ₂ O (0.1% TFA) Gradient 60/90 in 30 min; Rt: 9.01 min

R _I = CH ₂ Ph; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph)

ESI (+): [M + H] ⁺ = 406-408

HPLC: romasil Cl 8 CH ₃ CN / H ₂ O (0.1% TFA) 80/20; Rt: 6.23 min

R = CH ₂ (4-Ph-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph)

ESI (+): [M + H] ⁺ = 482-484

HPLC: ACE Cl 8 CH ₃ CN / H ₂ O (0.1% TFA) Gradient 60/90 in 30 min; Rt: 11.4 min

Method 2: The product IX c (2.03 mmol) is solubilized in TFA LOM in the presence of 5 eq of anisole (1.1 mL). The mixture is heated for 2.5 hours at 50 ° C. The solvents are evaporated under reduced pressure and the mixture is purified by semi-preparative HPLC to give compound I.

I b Rj = iBu; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph)

ESI (+): [M + H] ⁺ = 372-374

HPLC: ACE Cl 8 CH ₃ CN / H ₂ O (0.1% TFA) Gradient 60/90 in 30 min; Rt: 8.60 min Method 3: To a solution, under an inert atmosphere, of compound IX c (0.447 mmol, 120 mg) in 4.5 ml of TFA is added the thioanisole (2.8 eq, 0.936 mmol, 1 10 μΕ) followed by the addition of 2,2'-dithiobis (5-nitropyridine) (3eq, 1.0 mmol, 417 mg): the solution becomes orange. The mixture is stirred at ambient temperature for 1 h. The mixture is concentrated under pressure reduced to 30 ° C. The disulfide formed is purified by semi-preparative HPLC to give 146 mg of product (Yield: 62%).

The preceding disulfide is solubilized in Οί ^ Ν (720μΕ) / Η ₂ 0 (180μί). The tris (2-carboxyethyl) phosphine hydrochloride (1.2 eq; 96 mg) is then added and the mixture is stirred for 10 min at room temperature. The solvents are evaporated under reduced pressure and the compound is purified by semipréparative HPLC to give I b

This synthesis scheme makes it possible to preserve the chirality of the molecule if it exists

I b Ri = iBu; R ₂ = H; R ₃ = C¾ (4-Br-Ph)

ESI (+): [M + H] ⁺ = 372-374

HPLC: ACE C18 CH ₃ CN / H ₂ O (0.1% TFA) 60/90 over 30 min; Rt: 8.60 min

NMR (CDC1 _3, 200MHz): 0.87 (3H, d); 0.89 (3H, d); 1.50-1.80 (3H, m); 2.7-3.5 (6H, m); 7.08 (2H, d); 7.43 (2H, d)

Ig R = CH _3; R ₂ = CH ₃ ; R ₃ = CH ₂ (4-Br-Ph)

ESI (+): [M + Na] ⁺ = 366-368

HPLC: Atlantis T3 CH ₃ CN / H ₂ O (0.1% TFA) 50/90 in 10 min; Rt: 6.51 min

NMR (DMSO d6, 200MHz): 1.45 (3H, s); 1.47 (3H, s); 2.7-3.2 (5H, m); 7.18 (2H, d); 7.50 (2H, d)

R, = C ₂ H ₅ ; R ₂ = C ₂ H ₅ ; R ₃ = CH ₂ (4-Br-Ph)

ESI (+): [M + Na] ⁺ = 394-396

HPLC: Atlantis T3 CH ₃ CN / H ₂ O (0.1% TFA) 70/90 in 10 min; Rt: 3.9 min

NMR (DMSO d6, 200MHz): 1.30 (6H, t); 1.75 (4H, q); 2.6-3.1 (5H, m); 7.0 (2H, d); 7.40 (2H, d)

C (R] R ₂ ) = C ₄ H ₈ ; R ₃ = CH ₂ (4-Br-Ph)

HPLC: Atlantis T3 CH ₃ CN / H ₂ O (0.1% TFA) 60/90 in 10 min; Rt: 5.0 min

NMR (CDC1 _3, 200MHz): 1.60-2.25 (8H, m); 2.75-3.20 (5H, m); 7.10 (2H, d); 7.44 (2H, d); C (R, R ₂ ) = C ₅ H ₀ ; R3 = CH ₂ (4-Br-Ph)

ESI (+): [M + Na] ⁺ = 382-384

HPLC: Atlantis T3 CH ₃ CN / H ₂ O (0.1% TFA) 70/90 in 10 min; Rt: 8.97 min

NMR (CDCl ₃ , 200MHz): 1.60-2.40 (10H, m); 2.75-3.20 (5H, m); 7.10 (2H, d); 7.44 (2H, d) 1k C (R R ₂ ) = C ₈ H ₈ ; R ₃ = CH ₂ (4-Br-Ph)

HPLC: Atlantis T3 CH ₃ CN / H ₂ O (0.1% TFA) 70/90 in 10 min; Rt: 3.84 min

NMR (CDCl ₃ , 200MHz): 2.70 - 3.50 (5H, m); 3.60 (2H, d); 3.70 (2H, d); 7.10-7.6 (8H, m) C 1 (R ₂ ) = C ₅ H ₁₀ ; R ₃ = CH ₂ (4-Ph-Ph)

ESI (+): [M + H] ⁺ = 383

HPLC: Atlantis T3 CH ₃ CN / H ₂ O (0.1% TFA) 70/90 in 10 min; Rt: 5.35 min

NMR (CDC1 _3, 200MHz): 1.60-2.40 (10H, m); 2.75-3.20 (5H, m); 7.20-7.60 (9H, m)

I m C (RiR ₂ ) = C ₄ H ₈ ; R ₃ = CH ₂ (4-Br-Ph)

ESI (+): [M + H] ⁺ = 369

HPLC: Atlantis T3 CH ₃ CN / H ₂ O (0.1% TFA) 60/40; Rt: 9.79 min

NMR (CDCl ₃ , 200MHz): 1.60-2.25 (8H, m); 2.75-3.20 (5H, m); 7.05-7.55 (9H, m)

I n C (R 1 R ₂ ) = C ₈ H ₈ ; R ₃ = CH ₂ (4-Ph-Ph)

ESI (+): [M + H] ⁺ = 415

HPLC: Atlantis T3 CH ₃ CN / H ₂ O (0.1% TFA) 70/90 in 10 min; Rt .: 4.73 min

NMR (CDCl ₃ , 200MHz): 2.70 - 3.50 (5H, m); 3.60 (2H, d); 3.70 (2H, d); 7.10-7.8 (13H, m)

Measurement of inhibitory powers

The claimed inhibitors I have been tested on neprilysin type 1 (EC3.4.24.1 1, NEP-1). The assays are performed in 96-well plate in the presence of a specific fluorigenic substrate. In general, the inhibitors are pre-incubated in increasing concentrations with the enzyme for 10 min at 37 ° C. The substrate is then added and the mixture is incubated for 30 to 60 min at 37 ° C. The reaction is stopped at 4 ° C. and the reading of the fluorescence emitted is made in a Berthold Twinkle LS970B plate reader. An inhibition curve is then carried out as a function of the inhibitor concentration using the GraphPad software, then the Ki is determined from the Cheng Prusoff formula: Ki = IC ₅ o / (l + (S / Km) ). The results are presented in the following tables. Determination of Neprilysin (NEP)

Neprilysin, purified from rabbit kidney (Aubry M. et al., 1987, Biochem Cell Biol 65, 398-404), is used at 200 ng / ml final in 50 mM Tris buffer pH 7.4. The substrate, Dansyl-Gly- (NO ₂ ) Phe-B-Ala (Goudreau N. et al (1994) Anal Biochem., 219, 87-95) (Km = 37 μΜ), is dissolved in ethanol. and is used at 20μΜ final. Increasing concentrations (10 ^"10 to ^{10" 3} M) inhibitors were pre-incubated for 15 min at 37 ° C with the NEP-1 in 50 mM Tris buffer, pH 7.4. The substrate is then added and the incubation is continued for 60 minutes. The reaction is stopped by placing the plate in ice for 10 minutes. The reading of the fluorescence emitted is measured in a fluorimeter at 355 nm, λαη-535 nm.

Claims

claims

1. Compound of general formula (I)

RS-C (R, R ₂ ) -CO-CH ₂ -CH (R 1) -CO-R. ₍ (I)

in which:

a) R represents:

a hydrogen;

an alkoxyalkyloxycarbonyl R "C (O) O-CH (R" ') OC (O) - with R "and R"' representing, independently of one another, a linear or branched alkyl group, from 1 to 6 carbon atoms;

an S-cysteinyl (H ₂ N) (CO ₂ H) CH-CH ₂ -S- group; or

a group - ^ SC (R ₁ R ₂ ) -CO-CH 2 -CH (R ₃ ) -CO-R ₄ , with R _b R ₂ , R ₃ and R ₄ as below forming a compound of formula ( I) symmetrical;

b) Rj represents

a linear or branched alkyl group of 1 to 6 carbon atoms, substituted or not by:

^■ a group -OR _5, -SR 5 or -SO-R 5, with R ₅ representing a hydrogen, an alkyl group, linear or branched, 1 to 4 carbons, a phenyl or benzyl group;

^B a -C0 ₂ R6 group, with R ₆ representing a hydrogen, a linear or branched alkyl group comprising from 2 to 4 carbon atoms or a benzyl group;

^■ a group -NR ₇ R, with R ₇ and R representing, independently from each other hydrogen, an alkyl group, linear or branched, of 1 to 4 carbon atoms, a phenyl or benzyl group or -NR R ₇ taken together represent a saturated 5- or 6-membered ring comprising one or more heteroatoms selected from oxygen and nitrogen, preferably representing a morpholine or piperidine; ^■ a carboxamide group -CONR ₇ R 8, with -NR7R8 having the same definition as above;

^■ phenyl, substituted or not by one or more halogens selected from fluorine or bromine, or a group - OR _5, with R ₅ having the same definition as above;

A saturated 5- or 6-membered saturated cyclic or saturated 5- or 6-membered heterocyclic compound comprising 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur;

a saturated 5- or 6-membered saturated cyclic or saturated 5- or 6-membered heterocyclic compound comprising 1 or 2 heteroatoms chosen from oxygen, nitrogen and sulfur;

a phenyl group, substituted or not, by one or more halogens taken from fluorine or bromine, or by a group-OR ₅ , with R ₅ having the same definition as above;

and R ₂ is hydrogen; or

-C (RjR ₂ ) - taken together represents:

^H a 5-membered saturated cyclic compound, attached or not to an aromatic ring, preferably leading to an indanyl ring;

^B a 6-membered saturated cyclic compound;

3 represents:

a linear or branched alkyl group of 1 to 6 carbon atoms, substituted or not by: ^B a group -OR ₅ or -SR ₅ or -S (O) R ₅ , with R ₅ representing a hydrogen, a linear or branched alkyl group of 1 to 4 carbons, a phenyl or benzyl group;

^B a -CO ₂ R _{6 group,} with R ₆ representing a hydrogen, a linear or branched alkyl group comprising from 2 to 4 carbon atoms or a benzyl group;

^B a -NR7R8 group, with R ₇ and Rs representing, independently of one another, a hydrogen, a linear or branched alkyl group of 1 to 4 carbon atoms, a phenyl or benzyl group or with -NR ₇ Rg taken together representing a saturated 5- or 6-membered heterocycle comprising one or more heteroatoms selected from oxygen and nitrogen, preferably representing a morpholine or piperidine;

^H a carboxamide group -CONR ₇ R8 _; with -NR ₇ Rg as defined above;

^B a phenyl group substituted or not by:

one or more halogens selected from fluorine or bromine;

a group-OR5, with R having the same definition as above;

a phenyl or thienyl group;

B a 5- or 6-membered aromatic heterocycle comprising 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur; or "a 5- or 6-membered saturated 5- or 6-membered saturated or saturated heterocyclic ring compound having 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur;

a phenyl group substituted or not by:

^B one or more halogens selected from fluorine or bromine; or B a group -OR5 with R5 having the same definition as above;

B phenyl;

d) R4 represents:

a hydroxyl group; a group -NR7R.8, with R ₇ and Rs being, independently of one another, a hydrogen, a linear or branched alkyl group of 1 to 4 carbon atoms, a phenyl or benzyl group, or with NR ₇ Rg taken together representing a 5- or 6-membered heterocycle, having one or more heteroatoms selected from N or O;

an alkoxy group -OR ₉ , with R ₉ representing:

^■ an alkyl, linear or branched, having from 2 to 6 carbon atoms;

^■ a benzyl group;

^B a group -CHRio-COORn,

-CHR10 or -C (= O) -ORn in which Rio and Rn represent, independently of one another, a linear or branched alkyl group of 1 to 6 carbon atoms.

2. Compound according to claim 1 wherein Rj represents a linear or branched alkyl group of 1 to 6 carbon atoms or a linear or branched alkyl group of 1 to 6 carbon atoms substituted with:

B a phenyl group;

^■ a saturated cyclic compound of 5 or 6 membered heterocyclic or saturated 5- or 6-membered ring comprising 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur;

and R1 represents hydrogen, or

-C (Ri R ₂ ) - taken together represents:

^H a cyclic compound saturated 5-membered ring,

^■ a cyclic compound saturated 5-membered ring fused to an aromatic ring;

^■ a cyclic compound saturated 6-membered ring; or ^B a saturated 6-membered heterocyclic compound having 1 heteroatom, at the 4-position, selected from oxygen, nitrogen and sulfur;

A compound according to claim 1 or 2 wherein R3 represents a linear or branched alkyl group of 1 to 6 carbon atoms substituted with:

B a phenyl group; or

A phenyl group substituted with:

one or more halogens taken from fluorine or bromine; a phenyl or thienyl group.

Compound according to one of the preceding claims wherein R ₄ represents a hydroxyl.

Compound according to one of the preceding claims, in which:

a) R represents a hydrogen;

b) R 1 represents a linear or branched alkyl group of 1 to 6 carbon atoms or a linear or branched alkyl group of 1 to 6 carbon atoms substituted with:

^B a phenyl group;

^{H is} a saturated 5- or 6-membered saturated cyclic or saturated 5- or 6-membered heterocyclic compound having 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur;

and R2 is hydrogen, or

Ri and R are identical and represent a linear or branched alkyl group of 1 to 6 carbon atoms; or

-C (RjR2) - taken together represents:

^■ a cyclic compound saturated 5-membered ring, ^■ a cyclic compound saturated 5-membered ring fused to an aromatic ring;

A saturated 6-membered cyclic compound; or

^■ a heterocyclic compound saturated 6-membered ring having 1 heteroatom, in 4-position, selected from oxygen, nitrogen and sulfur;

c) R ₃ represents a linear or branched alkyl group of 1 to 6 carbon atoms substituted with:

^B a phenyl group; or

^■ a phenyl group substituted by:

one or more halogens selected from fluorine or bromine;

a phenyl or thienyl group;

d) R ₄ represents a hydroxyl group.

Compound according to one of the preceding claims wherein R] represents an isobutyl group or a methyl group substituted with:

B a phenyl group;

^■ a phenyl group substituted in position 4 by halogen selected from fluorine or bromine;

A phenyl group substituted in the 4-position by a phenyl group;

and R ₂ represents a hydrogen, or

R 1 and R ₂ are identical and represent a methyl or ethyl group, or

-C (R] R ₂ ) - taken together represents:

^■ a cyclic moiety, saturated 5 or 6 membered ring; or

^B a 5-membered saturated cyclic group attached to an aromatic ring.

Compound according to one of the preceding claims, in which R ₃ represents a carbon-substituted alkyl group substituted with:

H a phenyl group;

^H a phenyl group substituted in the 4-position by a halogen taken from fluorine or bromine; a phenyl group substituted in the 4-position by a phenyl group.

Compound according to one of the preceding claims, in which:

- R = CH ₂ Ph; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- R = CH ₂ Ph; R ₂ = H; R ₃ = CH ₂ Ph; or

- R = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Ph-Ph); or

- R = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ Ph; or

- R = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- R = CH ₂ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ Ph; or

- R = CH ₂ (4-Br-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- R = CH ₂ (4-Ph-Ph); R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- R = CH ₃ ; R ₂ = CH ₃ ; R ₃ = CH ₂ (4-Br-Ph); or

- R = C ₂ H ₅ ; R ₂ = C ₂ H ₅ ; R ₃ = CH ₂ (4-Br-Ph); or

- R -R ₂ = Cyclopentyl; R ₃ = CH ₂ (4-Br-Ph); or

- C R1R2) - = Cyclohexyl; R ₃ = CH ₂ (4-Br-Ph); or

- C R R ₂ ) - = Phenylcyclopentyl (Indanyl); R ₃ = CH ₂ (4-Br-Ph); or

- C R R ₂ ) - = Cyclohexyl; R ₃ = CH ₂ (4-Ph-Ph); or

- C R1R2) - = Cyclopentyl; R ₃ = CH ₂ (4-Ph-Ph); or

- c RjR ₂ ) - = Phenylcyclopentyl (Indanyl); R ₃ = CH ₂ (4-Ph-Ph).

9. Compound according to one of the preceding claims wherein:

- R 1 = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Ph-Ph); or

- R = CH ₂ CH (CH ₃ ) ₂ ; R ₂ = H; R ₃ = CH ₂ (4-Br-Ph); or

- C (RiR ₂ ) - = Cyclohexyl; R ₃ = CH ₂ (4-Br-Ph); or

- C (RiR ₂ ) - = Cyclohexyl; R ₃ = CH ₂ (4-Ph-Ph).

10. Compound according to one of the preceding claims wherein R is hydrogen and R4 is a hydroxyl group.

11. Compound according to one of the preceding claims for use as medicaments.

12. Compound according to claim 11 for its use as an antidiarrheal, antisecretory, anxiolytic, antidepressant, reductive cicatrizing of fluid effusions or in the treatment of cardiac water overload, oedemas and glaucoma.

13. Pharmaceutical composition comprising at least one compound according to one of claims 1 to 10 and at least one pharmaceutically acceptable excipient.

14. Pharmaceutical composition according to claim 13 for its use as antidiarrheal, antisecretory, anxiolytic, antidepressant, cicatrizant, reducing fluid effusions or in the treatment of cardiac water overload, oedemas and glaucoma.

15. The pharmaceutical composition of claim 13 or 14 for parenteral, oral, topical or nasal administration.

16. Cosmetic composition comprising at least one compound according to one of claims 1 to 10 and an acceptable excipient.