FR2778564A1 - Inhibition of apoptosis using a benzodiazepine peripheral receptor ligand - Google Patents

Abstract

The use of a ligand for benzodiazepine peripheral receptor in compositions to inhibit apoptosis. The ligands are preferably those compounds described in the following patents EP94271;EP210084;EP248734;EP295161;EP446141;FR2669926;EP524846;WO96/32 383;JP07165721;US4898861;EP346207;EP346208. Specific compounds that may be used are PK 11195, PK 14105, SR26276, Ro 5-4864, diazepam, and flunitrazepam, the formulae for which are given in the Figure. The use of SR26276 is especially advantageous.

Description

The present invention relates to the use of certain compounds and

  their pharmaceutically acceptable salts and solvates for the preparation of medicaments capable of

  reduce programmed cell death or apoptosis.

  The term apoptosis indicates all of the physiological processes linked to cell death. In its terminal phase, apoptosis is characterized by activation of endonucleases which cleave double stranded DNA in intemucleosomal regions thus generating mono and oligonucleosomes which complex with histones. We then observe in the cytoplasm of apoptotic cells an enrichment in oligo

  and histone-linked mononucleosomes.

  Although this phenomenon, unlike necrosis, is physiological, it can be

  also caused by pathological stimulation.

  D.A. Carson reports (The Lancet 1993, 341, 1251-1254) the role of apoptosis in certain pathologies such as immunosuppression, immune deficiencies in

  AIDS patients, cell aging, degenerative disorders.

  Mathieu. J. et ai. (Ann. Pharmaceutiques Français 1996, 54, 5, 193-201) have shown that the pathological effects caused by chemical and physical agents,

  as free radicals and ionizing radiation are caused by the pro-

apoptotics of these agents.

  Patent application WO 93/11771 claims a method for treating damage caused to the central nervous system by a head trauma or by cerebral ischemia using an agonist inhibitor of the peripheral benzodiazepine receptor. Apoptosis regulating products have been described in patent application WO 96/21449. The general formula includes both inhibitors and stimulators of

  apoptosis, without being given the means to distinguish from each other.

  It has now been found according to the present invention, that compounds having an affinity for the peripheral benzodiazepine receptor (PBR) have

  also anti-apoptotic activity.

  It was completely unexpected that receptors for benzodiazepines were discovered on the kidney (Braestrup. C. et al., Proc. Natl. Aca. Sci. USA 1977, 74, 3805-3809) and then on many other "peripheral" tissues such as heart, lung, adrenal glands, hence their name peripheral benzodiazepine receptors (PBR). This class of binding sites is physically and pharmacologically distinct from the "central" benzodiazepine receptors (CBR). Indeed, the most powerful benzodiazepines therapeutically have a low affinity for the peripheral site. Conversely, the affinity of certain benzodiazepines devoid of anxiolytic effect is 100 to 1000 times stronger for the peripheral receptor than for the central receptor. It is on this property that the pharmacological distinction of the two classes of receptors is based. Thus, ligand-receptor interaction studies (binding assay) show that the PBR specifically fixes the tritiated PK 11195 or the tritiated Ro 5-4864 with a dissociation constant of the order of nanomolar. The binding of these two radioligands is displaced by the following ligands, classified according to an order of decreasing displacement power: PK 11195> Ro 5-4864> diazepam = flunitrazepam >> clonazepam (Le Fur. G. et al., Life Sci. 1983, 32, 1839-1847 and 1849-1856; Wang. JKT et al., Mol. Pharmacol. 1984, 25, 349-351). Clonazepam is not recognized by PBR, it is a selective ligand for the central receptor. Diazepam and flunitrazepam have an equivalent affinity for the two types of receptors, they are said to be "mixed". The PK

  11195 and Ro 5-4864 only bind to peripheral type receivers; the Ro 5-

  4864 presenting the thermodynamic characteristics of an agonist and PK 11195 those of an antagonist (Le Fur. G. et al., Life. Sci. 1983, 33, 449-457). It should be noted that these ligands are classified into two structural groups. The first includes the benzodiazepines themselves: Ro 5-4864; diazepam, flunitrazepam, clonazepam while the second is formed from isoquinoline carboxamide derivatives: PK 11195, PK 14105, SR 26276 (Figure 1). Finally, PK 14105 and flunitrazepam have a particular property: under ultraviolet irradiation, their nitro substitution is activated and forms a covalent bond with an amino acid

  close. This property was used for the characterization of the two receptors.

  If the pharmacological characterization makes it possible to distinguish the two types of receptors, central and peripheral, other properties also contribute to it. Despite their name, peripheral sites also exist in the central nervous system, but their distribution is different from that of central sites; they are preferentially located on the glial cells rather than on the neurons. Conversely, CBR is practically absent from peripheral tissues, while PBR is widely distributed throughout the body. The latter is detected in large quantities in endocrine function tissues (adrenal glands, testes and pituitary gland) as well as in tissues with rapid cell renewal (skin, lingual and nasal epithelium). Its presence becomes weaker in smooth muscles and tissues of the digestive tract. A certain heterogeneity is found in the lymphoid organs studied by the autoradiography technique on sections with the ligand PK 11195 tritiated: the highest density of sites is detected in the lymph nodes followed by intestinal Peyer's plaques, the spleen and the thymus . Inside each organ, the labeling suggests a preferential distribution at the level of T lymphocytes and macrophages. Particularly rich areas are the marginal area of the white pulp for the spleen, the medulla for the thymus, the paracortex of the lymph nodes and the periphery of the follicles for plaques

of Peyer.

  At the subcellular level, the peripheral receptor seems to be mainly associated with mitochondria as indicated by several correlations obtained between the binding activity of specific PBR ligands and the specific enzymatic activity of the mitochondria, after subcellular fractionations. More specifically, it would be a protein of 17-18 kd of molecular mass, localized in the outer membrane of the mitochondria (Anholt. R; R; H. et aI, J. Biol. Chem. 1986, 261, 576 -583; Doble. A. et ai, Mol. Pharmacol. 1987.31, 42-49; Antkiewicz-Michaluk. L. et al, Mol.

  Pharmacol. 1988, 34, 272-278; Hirsch. J. D. et al, Mol. Pharmacol. 1988, 34, 164-172).

  However, a membrane localization of the PBR was detected in the nuclear fraction (Marangos. PJ et al, Mol. Pharmacol. 1982, 22, 2633; Anholt. RRH et al, J. Biol. Chem. 1986, 261, 576-583 ) or on membrane structures (Doble. A. et al, Eur. J. Pharmacol. 1985, 119, 153-167; Antkiewicz-Michaluk et al, Mol.

Pharmacol., 1988, 34, 272-278 1988).

  At the nucleic level, the cDNA coding for PBR has been cloned and sequenced. This cDNA codes for a protein of 169 amino acids, devoid of cysteine residue, having a predominance of basic amino acids and having 5 hydrophobic transmembrane regions. The most likely conformation and orientation of the protein

  would be given by these hydrophobic domains brought together and housed in the membrane.

  In terms of expression, Northern blot analyzes demonstrate that the major quantitative differences in the levels of PBR-specific mRNA in different tissues are comparable to the differences in density of the PBR itself, found in these same tissues. This suggests that the cellular amount of PBR is regulated at the level of gene expression. The use of the human PBR cDNA as a probe made it possible to localize the PBR gene in the 22q13.3 band of

human genome.

  The absence of homology between the sequence of the PBR and that of the various subunits making up the CBR confirms a distinction drawn on the pharmacology and the

tissue localization.

  Finally, unlike the central benzodiazepines for which the discovery of a pharmacological role preceded the characterization of a receptor, the peripheral site was detected in the absence of significant effects of these same benzodiazepines at the periphery, o a first qualification of "acceptor". But currently, the term "receiver" seems to be justified due to many effects

  biological effects induced by the binding of a ligand.

  The exact function of the PBR receptor remains unknown to date, but an overexpression of this receptor has been found during many phenomena consecutive to ischemia (cerebral in particular), of traumatic origin or during degeneration processes (neural types Alzheimer, amyotrophic lateral sclerosis or

  ALS ...). Most of the time, this overexpression of PBR, measured by binding of 3H-

  PK 14105, was considered to be a cellular activation factor (linked to

presence of macrophages).

  The natural ligands of PBR are poorly understood. Protoporphyrins (IX in particular) appear to be reasonable candidates but it has never been implemented

  evidence of any pharmacological effect of these products.

  The intracellular location, at the mitochondrial level of PBR is well known and could be responsible for the relationship with early apoptosis processes at the level of regulation of mitochondrial oxidative metabolism. In this regard, reference can be made to Carayon's publication. P. et ai., Blood 1996, 87, 3170-3178, which suggested that certain cells of the hematopoietic system could be resistant to oxidative stress by overexpression of bcl2 (a well-known anti-apoptotic protein), in close relation with level of expression of PBR. To date, it has never been demonstrated that PBR ligands can have a pro- or anti-apoptotic effect in any system having any pharmacological effect in pathologies linked to a dysregulation of apoptosis. These disturbances are also

many.

  Thus, the present invention relates to the use of PBR ligands as compounds

  anti-apoptotics as well as pharmaceutical compositions containing them.

  The anti-apoptotic activity of these compounds was measured on muscle cells

  human aortic smooths on a lymphoblastoid cell line (U937) opposite

  vis the pro-apoptotic activity of a deprivation in serum or after the addition of toxic compounds such as tumor necrosis factor a (TNFa) or hydrogen peroxide (H202) using a dosing kit ELISA (from the English "enzyme-linked immunosorbent assay) specific detecting the presence of oligonucleosomes whose existence inside the cells is a specific marker of programmed cell death (apoptosis), according to the method described by Del Bino G. et ai., Experimental Cell Research 1991, 27, 193 and 1991, 195, 485 or Darzynkiewicz A et ai., Cytometry

1992, 13, 795.

  Advantageous PBR ligands used in the pharmaceutical compositions of the present invention are those of formulas (1) to (XIII) below: I - the compounds of general formula (1): R Ci / ABy CONjR (I Ai Ar, in which: - R11 and R22 independently represent an alkyl group comprising 1 to 6 carbon atoms, linear or branched, a cycloalkyl group comprising 3 to 7 carbon atoms, a phenylalkyl or cycloalkylalkyl group, the alkyl part of which comprises 1 to 3 atoms carbon, - R11 and R21 may also represent an alkenyl or alkynyl group comprising 3 to 6 carbon atoms, provided that the double or triple bond is not located in position 1-2 relative to the nitrogen atom , - R. 'and R21 may also represent a group of formula -R31-Z1-R41 in which R31 represents a linear or branched alkylene group of 2 to 6 carbon atoms, provided that at least 2 carbon atoms separate the nitrogen of the grou eg Zl, - R41 represents an alkyl group comprising from 1 to 4 carbon atoms and Z1 the oxygen or sulfur atom or the group> N-R51, Rs1 representing the hydrogen atom or an alkyl group of 1 with 3 carbon atoms, - R11 and R21 can form with the nitrogen atom to which they are attached a heterocycle; optionally comprising a second heteroatom, - Ar, represents a phenyl, pyridyl or thienyl group or a phenyl group substituted by one or two substituents chosen from halogen atoms, alkyl, alkoxy and alkylthio groups having 1 to 4 carbon atoms, the trifluoromethyl group and the nitro group; - A1 and B, independently represents N or -CH CLv represents the sequence or when A1 represents N and Yi B1 represents CH, one of the following sequences:

I s-

  - XI and Y1 independently representing the hydrogen atom, a halogen atom, an alkyl or alkoxy group comprising 1 to 3 carbon atoms, the nitro or trifluoromethyl group described in EP 94 271; II - the compounds of general formula (II): V2 X-X (CH2) r-- (C H) m-2 CO-N \ / A Rj2 2

W 2 Z2

  in which: - A2 represents a nitrogen atom or a CH group, - B2 represents a nitrogen atom or a CH group, - V2 and W2 identical or different, represent hydrogen or halogen atoms, groups alkyl or alkoxy having 1 to 3 carbon atoms, nitro or trifluoromethyl groups, - Z2 is fixed in the ortho or para position with respect to B2 and represents a phenyl, thienyl, pyridyl or phenyl radical substituted by one or two substituents taken from the halogen atoms, alkyl and alkoxy groups containing 1 to 4 carbon atoms, trifluoromethyl or nitro groups, - the chain X2- (CH2) n2 - (CHR2) m2 -CO-NR, 2R22 is fixed in the ortho position or para with respect to B2, - R2 represents a hydrogen atom or an alkyl group having 1 to 3 carbon atoms, - R12 and R22, identical or different, represent a linear or branched alkyl group containing 1 to 6 carbon atoms, cycloalkyl having 3 to 6 carbon atoms, p hényle, phenylalkyle ou cycloalkylalkyle of which the alkyl part contains 1 to 3 carbon atoms and the cycloalkyle part contains 3 to 6 carbon atoms, alkenyl containing 3 to 6 carbon atoms provided that the double bond is not located in position 1, 2 with respect to the nitrogen atom, - R12 and R22 can also form, together with the nitrogen atom to which they are attached, a pyrrolidine, piperidine, morpholine or thiomorpholine ring, - X2 represents a group CH- R32, N -R42, SO, SO2 or an oxygen or sulfur atom, - R32 represents a hydrogen atom or an alkyl group having 1 to 3 carbon atoms, - R42 represents an alkyl group having 1 to 3 carbon atoms, - m2 is equal to 0 or 1 - n2 is equal to 0, 1 or 2, it being understood that when X2 represents a group SO, SO2 or N-R42 the sum m2 + n2 is at least equal to 1, it being understood that when A2 and B2 each represent a nitrogen atom and Z2 is in the para position relative to port to B2, X2 cannot represent the CH-R32 group, it being understood that when A2 represents a CH group, B2 represents a nitrogen atom, Z2 is in the ortho position with respect to B2, X2 represents an oxygen atom and R2 represents a hydrogen atom, the sum m2 + n2 is different from 1 and with the exception of N, N-dimethylcarbamate of phenyl-2-quinolyl-4, its diastereoisomers, racemic and enantiomers and when it can exist , its addition salt with an acid described in EP 210 084; III - the compounds of general formula (III): R CO-N

X R3

2C (111)

  Ar3 in which: - Ri3 and R23, identical or different, represent straight or branched chain alkyl groups containing 1 to 4 carbon atoms, cycloalkylalkyl in which the alkyl part contains 1 to 3 carbon atoms and the cycloalkyl part comprises 3 to 6 carbon or phenyl atoms, - R13 and R23 can also form together with the nitrogen atom to which they are attached a piperidine ring, - Ar3 represents a phenyl, thienyl or phenyl radical substituted by one or two substituents chosen from atoms 'halogen, alkyl or alkoxy groups containing 1 to 4 carbon atoms, nitro or trifluoromethyl and - X3 <represents one of the following sequences:

us o-

  1tL.s, [XX, and when the group NR13R23 comprises at least one asymmetric carbon atom, their racemates and stereoisomers described in EP 248 734; IV - the compounds, in the form of pure enantiomers or of a mixture of enantiomers, corresponding to the general formula (IV): N

N / X4

  R1N / ofeS4 in which, - X4 represents a hydrogen or halogen atom, a C1-C4 alkoxy group or a C1-C6 alkyl group, - Y4 represents a hydrogen atom or a methyl group, - R , 4 represents a C1-C4 alkyl group, and - R24 represents a C1-C6 alkyl group, as well as their addition salts with acids acceptable in pharmacology described in

EP 295,161

  V - imidazo [1,2-c] quinazoline derivatives of general formula (V): X5

R3N N N (V)

  R5 in which: -Y5 represents an oxygen or sulfur atom; -R15 represents: a) a radical having 1 to 6 carbon atoms in a straight or branched chain,

  optionally substituted by a phenyl radical itself optionally mono- or di-

  substituted by a halogen atom or an alkyl or alkoxy radical each having from 1 to 6 carbon atoms in a straight or branched chain; b) a mono, bi or tricyclic cycloalkyl radical containing from 1 to 3 carbon atoms; c) an aromatic group chosen from the group formed by:

  - an unsubstituted phenyl radical and the mono- and di- phenyl radicals

  substituted by a substituent chosen from halogen atoms and alkyl and alkoxy radicals each having from 1 to 6 carbon atoms in a straight and branched chain, and - a furyl radical and a thienyl radical, or d) an acyl radical, such that for example an alkoxycarbonyl, aminocarbonyl, N, N-dialkylaminocarbonyl radical in which the alkyl group contains from 1 to 6 carbon atoms in a straight or branched chain, or a benzoyl radical optionally mono- or di-substituted by a halogen atom or an alkyl or alkoxy radical each having from 1 to 6 carbon atoms in a straight or branched chain; - R25 represents a hydrogen or halogen atom or an alkyl radical containing from 1 to 6 carbon atoms optionally substituted by an amino alkylamino or dialkyl amino radical in which the alkyl part contains from 1 to 6 carbon atoms in a straight chain or branched; - R3s represents: a) a hydrogen atom, b) an alkyl radical having from 1 to 6 carbon atoms in a straight or branched chain, optionally substituted by an aryl radical such as for example a phenyl radical, itself optionally mono- or poly-substituted by an alkyl or alkoxy radical each having from 1 to 6 carbon atoms in a straight or branched chain, or; c) a radical of formula:

R5-CO-A5-

  in which: - A5 is a straight or branched chain alkylene radical having from 1 to 6 carbon atoms, and - R5 is an alkoxy radical having from 1 to 6 carbon atoms or an amino radical of formula: -NR "'in which R'5 and R "5, identical or different, each represent a hydrogen atom or an alkyl radical containing from 1 to 6 carbon atoms in a straight or branched chain optionally substituted by a hydroxy or alkoxy radical having from 1 to 6 carbon atoms, or R'5 and R "s form together with the nitrogen atom to which they are linked, a penta or hexagonal heterocyclic radical optionally containing a second heteroatom and optionally substituted such as for example a morpholino or N- radical methylpiperazinyl, d) an aryl radical such as for example a phenyl radical, optionally substituted by one or more alkyl or alkoxy radicals each having from 1 to 6 carbon atoms in a straight or branched chain, - X5 represents a hy atom drogen, described in EP 446 141 VI compounds of general formula (VI): R6 z6 6

X6 I I 11

  N-C-N_ (vI) R 6 in which, - X6 and X'6 represent a hydrogen or a halogen, X'6 possibly being in position

ortho, meta or para.

  - R6 is hydrogen or a C1 to C3 alkyl - Z. is an oxygen or sulfur atom R16 is a C1 to C5 alkyl and R26 is a phenyl group optionally substituted by a nitro group or a halogen, or well R16 and R26 together form a heterocycloalkyl with at least one heteroatom constituted by the nitrogen atom to which they are attached described in FR 2 669 926 VII - the compounds, in the form of pure optical isomer or mixture of two enantiomers , corresponding to the general formula (VII): 7 \ OH A / y7 ó (

M A I N (VII)

  N in which: - R17 represents a hydrogen or halogen atom or a methyl group, - R27 represents one or two substituents chosen from hydrogen and halogen atoms and (C1-4) alkyl, hydroxy groups , (C1-4) alkoxy, trifluoromethyl, acetylamino and methylsulfonylamino, and either X7 represents a CH2 group and Y7 represents a CH2 group; (CH2) 2 or CO, or X7 represents a CO group and Y7 represents a CH2 group, being excluded the compound in the formula of which R17 and R27 each represent a hydrogen atom and X7 and Y7 each represent a CH2 group, to the state of free base or of pharmaceutically acceptable acid addition salt described in EP 524 846; VIII - acetamide derivatives of general formula (VIII):

8 R3 R 8

R7, / R1

xi N "

R8 N 0 R2

  8a> \ / N- NR (Viii) in which: - X8 represents an oxygen atom or a group NR4, - R38 represents a hydrogen atom, an alkyl or hydroxylalkyl group, - R48 represents a hydrogen atom or an alkyl group, or

  - R38 + R48 represents a complete pyrrolidine, piperidine or a 2,3-dihydro- group

  1 H-indole, - R58 represents a hydrogen atom, an alkyl group (optionally substituted by a hydroxyl, an amino optionally itself protected by a carboxyl which can be substituted, benzyloxy, acyloxy or alkoxy), alkenyl, CF3, halo, amino, monoalkylamino, dialkylamino, acylamino, nitro, carbamoyl, mono (alkyl) carbamoyl, di (alkyl) carbamoyl or optionally protected carboxyl, - R68 represents a hydrogen atom, an optionally substituted alkyl, trifluoromethyl or phenyl group, or - R38 + R68 represents a group (CH2)., N8 = 3 to 6, - R78 represents a hydrogen, a halogen, an alkyl or alkoxy group, CF3, hydroxy, cyano, nitro, amino, monoalkylamino or dialkylamino, - R88 represents a hydrogen , a halogen, an alkyl or alkoxy group, - RI8 represents a hydrogen, an alkyl, alkenyl or cycloalkylalkyl, - R28 represents an alkyl, cycloalkyl group, an optionally substituted phenyl or optionally substituted phenylalkyl é or, -NR18R28 represents a group of formula:

--N A8

  R "in which: - A8 corresponds to a bond, methyl, oxygen or HN, R'8, R" 8 represents hydrogen or alkyl, and if A8 represents a bond, and R'8 and R "8 are in position 2 and 3 on the cycle, then R'8 and R "8 can merge and form a benzene cycle described in patent application WO 96/32383; IX- isoquinolinecarboxamide derivatives of general formula (IX): R C-N R2

C N R

N (IX)

  j-- X9 in which: - R91 and R92 represent an alkyl group comprising from 1 to 6 cycloalkyl carbon atoms comprising from 3 to 7 phenylalkyl carbon atoms with 1 to 3 carbon atoms or (C3C7) cycloalkyl (C1-C3) alkyl, and - X9 represents an iodine or a bromine described in JP 07165721 X - the N-phenylthiourea derivatives of general formula (X): 1o N - C - Np, 0 (x; 0 RR x'0 in which: - X10 and X'10 represents a hydrogen or a halogen, - R represents a hydrogen or a (Cl-C3) alkyl group, - Zl0 represents an oxygen or sulfur atom, R 10 represents an alkyl group comprising from 1 to 5 carbon atoms, R2 10 represents a phenyl group optionally substituted by a halogen atom or a nitro group, - NR1t R2 'comprises at least one heteroatom consisting of the nitrogen atom described in FR 2 669 926; XI the compounds of general formula (XI): il P v N Xi 1 "-N (X1) Y il in which: - R.1" represents a (Có-C3) alkyl group, (C 2- C3) alkenyl or (C2-C3) alkynyl, - Y ,, represents a halogen, hydroxy or alkoxy, - X ,, represents halogen or a hydrogen and, - Zl represents a hydrogen atom or a halogen described in US 4,898,861 A XII - the compounds of general formula (XII):

12 / R

I'4-N- (CH2) n, 2 CO-N \ 12

12,212

A'12 CO2R3 R

12 B (XII)

  d1 t> DAN in which: - R, '2 and R22, independently of one another, represent the hydrogen atom, a C1 to Cs alkyl group, a phenyl or benzyl group, or R1'2 and R2'2 can form, with the nitrogen atom to which they are attached, a saturated heterocycle C4 to C8, - R312 and R412, independently of each other, represent the hydrogen atom or an alkyl group in C1 to C6, -R512 represents a hydrogen or halogen atom, or a C, to C4 alkyl, C1 to C4 alkoxy, nitro or trifluoromethyl group, it being understood that the alkyl groups may be linear, branched or cyclic, and that the phenyl or benzyl groups may be optionally substituted by C1 to C6 alkyl, C1 to C6 alkoxy, trifluoromethyl or nitro groups, or by halogen atoms, -nl2 is 1,2 or 3 and - one of the symbols A12, B12, C12 or D12 represents N and the others represent CH, - as well as the N-oxides of heterocyclic nitrogen and their ls of addition with pharmaceutically acceptable acids and bases described in patent EP 346 207B; XIII - the compounds of general formula (XIII): R13

13 I 13 13

  13 R4-N-CH- (CH2) n-CONR1R2 R, d Lc1o z y3 (II _13

1 \ A13 C0Z13

1> k3 (XIII)

R C.Z_ N R13

(to) P

  in which: - R113 and R213 identical or different, each represent the hydrogen atom, a C1 to C6 alkyl group or C2 to C6 alkenyl, a phenyl or benzyl group, or R113 and R213 form with the nitrogen atom to which they are attached a saturated heterocycle chosen from piperidine, pyrrolidine, morpholine and piperazine, - R313 represents the hydrogen atom, a C1 to C6 alkyl group, phenyl C7 to Cg alkyl, or phenyl - R413 represents the d atom hydrogen or a C1 to C4 alkyl group - Rs513 and R613 identical or different each represent a hydrogen or halogen atom, a C1 to C3 alkyl or alkoxy group, a nitro group, trifluoromethyl, or together form a methylenedioxy group , - Z13 represents OR13 in which R713 represents the hydrogen atom or a C1 to C6 alkyl group; NR8 13R9 13 wherein R813 and R913 each represent a hydrogen atom, a C1-C4 alkyl group, a phenyl or benzyl group; a C1-C4 alkyl group; a benzyl group; an aryl group chosen from phenyl, pyridyl, pyrrolyl, furyl, thienyl and imidazole groups, - R1013 represents the hydrogen atom, a C1-C4 alkyl group or a phenyl group, it being understood that when Z13 is not a benzyl or aryl group, R313 does not represent H, the phenyl and benzyl groups possibly being substituted by halogen atoms, C1 to C3 alkoxy, alkyl and thioalkyl groups, nitro, trifluoromethyl and hydroxy groups, alkyl groups and alkoxy can be linear, branched or cyclic, -n13 represents 0.1 or 2, represents 0 or 1 and - one of the symbols A13, B13, C13 D13 represents N and the other CH or A13, B13, C13, D13 represents each CH, in the form of a racemic or enantiomers, as well as their addition salts with pharmaceutically acceptable acids or bases described in patent EP 346,208; An advantageous compound for the use according to the invention is SR 26276 and its salts

  and solvates described in EP 346 208, example number 84.

  Representative compounds of the invention and their pharmaceutically acceptable salts, as a function of time and dose, inhibit the pro-apoptotic effect induced

  by deprivation in serum or by the addition of TNFa or H2O2.

  Thus, according to a further aspect, the present invention relates to the use of

  compounds of formula (I) to (XàI), of an advantageous or preferred compounds mentioned above

  above, or their pharmaceutically acceptable salts and solvates for the preparation of a medicament for the treatment of diseases which may be

  treated by a decrease in apoptosis.

  Such pathologies are, for example, diseases linked to abnormal apoptotic activity, ocular affections such as cataracts or glaucoma, osteoporosis, bone fractures, epidermal lesions, restenosis, conditions linked to incorrect proliferation or migration of cells. smooth muscles, inflammations of the respiratory system, asbestosis, silicosis, lupus erythematosus, granulomatosis, eosinophilic granulomatosis, gastric and duodenal ulcers, esophagitis, enteritis, gastritis, septicemia, dysfunctions of the hematopoiesis and / or lymphopoiesis, cystic fibrosis, psoriasis, peripheral neuropathies, brain damage caused by an attack, diseases causing demyelination in general such as multiple sclerosis, diseases which involve neuronal degeneration such as vascular dementia, disorders post-apoplectic, post-syndromes traumatic brain injury, cerebral anoxia disorders, Alzheimer's disease, senile dementia, subcortical dementia, such as Huntington's disease and Parkinson's disease, dementia caused by AIDS, neuropathies derived from morbidity or damage to the body sympathetic or sensory nerves, and brain diseases, such as cerebral edema, and spinocerebellar degeneration, amyotrophic lateral sclerosis, or diseases requiring cardioprotection such as myocardial infarction, cardiac ischemia, heart failure, vasospasm coronary artery disease, angina, heart disease and heart valve disease, inflammatory heart disease, side effects from medication

  cardiotoxics or in patients undergoing cardiac surgery.

  According to a particularly advantageous aspect, the present invention relates to the use of the compounds of formula (1) to (XIII) of one of the advantageous or preferred compounds mentioned above, or of their pharmaceutically acceptable salts and solvates

  for the preparation of medicaments capable of inhibiting apoptosis.

  It is thanks to this anti-apoptotic activity that the compounds of the present invention can be used for the preparation of medicaments intended for infections by antiviruses such as HIV and HTLV 1 and 2 (from the English human immunodeficiency virus and human T lymphocyte virus) and their consequences such as leukemia, myelopathies and arthropathies, hepatitis (C, A, B, F), AIDS, immune deficiencies, cell aging, phenomena of tissue degeneration, inflammation, cell proliferation, infectious diseases, transplant rejection, acute or chronic rheumatoid arthritis, ulcerative colitis, thrombocytopenic purpura, autoimmune hemolytic anemia, type I (insulin-dependent) diabetes, myelodysplastic syndrome, prion diseases, respiratory distress syndrome, prostatic hypertrophy, asthma, atherosclerosis and its complications thromboembolic disorders, kidney disease and glomerulonephritis, diseases related to abnormal apoptotic activity, eye conditions such as cataracts or glaucoma, osteoporosis, bone fractures, skin damage, restenosis, conditions related to improper proliferation or migration of smooth muscle cells, inflammation of the respiratory system, asbestosis, silicosis, lupus erythematosus, granulomatosis, eosinophilic granulomatosis, gastric and duodenal ulcers, esophagitis, enteritis, gastritis, septicemia , dysfunctions of hematopoiesis and / or lymphopoiesis, cystic fibrosis, psoriasis, peripheral neuropathies, brain damage caused by an attack, diseases causing demyelination in general such as multiple sclerosis, diseases which involve

  neuronal degeneration such as vascular dementia, post-

  apoplectics, post-traumatic brain injury syndromes, disorders of cerebral anoxia, Alzheimer's disease, senile dementia, subcortical dementia, such as Huntington's chorea and Parkinson's disease, dementia caused by AIDS, neuropathies derived of morbidity or damage to the sympathetic or sensory nerves, and brain diseases, such as cerebral edema, and spinocerebellar degeneration, amyotrophic lateral sclerosis, or diseases requiring cardioprotection such as myocardial infarction, cardiac ischemia, heart failure, coronary vasospasm, angina, heart disease and heart valve disease, inflammatory heart disease, side effects from cardiotoxic drugs or

  still in patients undergoing cardiac surgery.

  According to the aim of the present invention, by the expression "treatment of diseases" is meant

  includes both disease care and prevention, where possible.

  So, for example, when we consider transplant rejection, the compositions

  Pharmaceuticals can be used for prevention.

  According to another of its aspects, the present invention relates to a method for inhibiting apoptosis, which comprises administering to a mammal in need of an effective dose of a compound of formula (I) to (XII), d one of the advantageous or preferred compounds mentioned above, or of a salt and solvate thereof

pharmaceutically acceptable.

  According to a preferred aspect, SR 26276 and its salts and solvates pharmaceutically

  acceptable are administered in microparticulate form.

  The compounds of formula (I) to (XmI), one of the advantageous or preferred compounds mentioned above or their pharmaceutically acceptable salts and solvates are of

  preferably administered orally.

  The amount of active ingredient to be administered depends on the degree of progression of the disease and on the age and weight of the patient. Nevertheless, the unit doses generally comprise from 0.25 to 700 mg, advantageously from 0.5 to 300 mg, preferably from 1 to 150 mg, for example between 2 and 50 mg of active principle. These unit doses are normally administered one or more times a day, preferably one to three times a day, the overall dose in humans being variable between 0.5 and 1400 mg per day, for example from 1 to 900 mg per day. , advantageously from 2 to 500 mg per day, more suitably from 2 to 200 mg per day. When the active ingredient administered is for example SR 26276, the unit dose generally comprises from 0.5 to 10

  mg, advantageously from 1 to 5, preferably from 1 to 3 mg, for example 1 1.5 - 2 -

  2.5 - 3 mg of active ingredient. These unit doses are normally administered one or more times a day, preferably one to three times a day, the overall dose in humans being variable between 0.5 and 50 mg per day, for example from 1 to 20 mg per day. ,

  advantageously from 2 to 10 mg per day.

  The above doses and quantities refer to the compounds of formula (I) to (XIII) or to

  one of the advantageous or preferred compounds cited above, in nonsalified form.

  In the pharmaceutical compositions of the present invention for oral administration, the active principle can be administered in unit administration forms, in admixture with conventional pharmaceutical carriers, to mammals, animals and humans for the treatment of the abovementioned conditions . Suitable unit dosage forms include, for example, optionally scored tablets, capsules, powders, granules and

oral solutions or suspensions.

  When preparing a solid composition in the form of tablets, the main active ingredient is mixed with a pharmaceutical vehicle such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic or the like. The tablets can be coated with sucrose or other suitable materials or else they can be treated in such a way that they have a prolonged or delayed activity and that they continuously release a predetermined quantity of active principle. A preparation in capsules is obtained by mixing the active ingredient with a diluent

  and by pouring the mixture obtained into soft or hard capsules.

  A preparation in the form of a syrup or elixir may contain the active ingredient together with a sweetener, preferably calorie-free, methylparaben and propylparaben as antiseptics, as well as a flavoring agent and an appropriate color. Water dispersible powders or granules may contain the active ingredient in admixture with dispersing or wetting agents, or suspending agents, such as polyvinylpyrrolidone, as well as with

  sweeteners or flavor correctors.

  The active principle can also be formulated in the form of microcapsules,

  possibly with one or more supports or additives.

  In the pharmaceutical compositions according to the present invention, the active principle can also be in the form of an inclusion complex in cyclodextrins, their ethers or their esters. The PREPARATIONS and EXAMPLES below illustrate

the invention better.

PREPARATION 1

  000 smooth muscle cells isolated from the human aorta (supplier CLONETICS) are placed in a 35 mm dish in a medium containing 2 ml of DMEM (Eagle medium modified by Dulbecco containing 4.5 g / l of glucose, 3.7 g / I NaHCO3 and does not contain L-glutamine or Na-pyruvate). 20% v / v of fetal calf serum decomplemented for 30 minutes at 80 ° C., 4 mM L-glutamine, U / ml of penicillin and 50 μg / ml of streptomycin are added. The cells are left in this medium for a growth period of three days before being subjected to

  experiments according to the examples reported below.

PREPARATION 2

  000 cells of the U937 lymphoblastoid line (supplier: ATCC) are placed in a 35 mm dish in a medium containing 2 ml of RPMI 1640 (containing 2 g / l of glucose, and not containing L-glutamine). 20% v / v of fetal calf serum decomplemented at 80 ° C., 4 mM L-glutamine, 50 U / ml of penicillin and 50 μg / ml of streptomycin are added. The cells are left in this medium for a growth period of three days before subjecting them to the experiments.

  according to the examples reported below.

PREPARATION 3

  The boxes containing the cells are prepared as in PREPARATION 1 or preparation 2. Apoptosis is induced by three different methods: a) by replacing the medium of PREPARATION 1 with the same medium containing

than 0.2% fetal calf serum.

  b) by adding to the medium described in PREPARATION 1 increasing doses of TNFa (0.01 ng / ml to 100 ng / ml) c) by adding to the medium described in PREPARATION 1 or PREPARATION 2 increasing doses of H2O2 ( 0.11PM to 100PM) By an ELISA test for assaying the mono- and oligonucleosomes associated with cytoplasmic histones after washing and cell lysis, the effects of the compounds of the invention on apoptosis are measured after 24 hours of contact with the cells in comparison with the rates of apoptosis obtained in the absence of products (maximum rate of apoptosis) or in the presence of 20% of fetal calf serum (rate

minimum of apoptosis).

EXAMPLE 1

  Boxes containing cells are prepared as in PREPARATIONS 1 or 2. Boxes containing cells are prepared as in PREPARATION 1 or 2 and apoptosis is induced as in PREPARATION 3 according to method a). The anti-apoptotic effects of SR 26276 and of compound Ro5-4864 are measured after 24 hours of contact with the cells in comparison with the controls (0.2% fetal calf serum and 20% fetal calf serum

  b) by the method described in PREPARATION 3.

  The compounds tested significantly inhibit apoptosis induced by deprivation of serum after 24 hours of contact with the cells and for at least 7 days.

EXAMPLE 2

  Boxes containing cells are prepared as in PREPARATION 1 or 2. Apoptosis is induced according to method b) of PREPARATION 3. The levels of apoptosis are measured after 24 hours of contact with the cells by the method described in PREPARATION 3; in comparison with the controls (20% fetal calf serum and 20% fetal calf serum + TNF alpha) by the method described in PREPARATION 3. SR 26276 and the compounds Ro 5-4864 significantly inhibit

  significant the pro-apoptotic effect of TNFo.

EXAMPLE 3

  Boxes containing cells are prepared as in PREPARATION 1 or 2. Apoptosis is induced according to method c) of PREPARATION 3. The levels of apoptosis are measured after 24 hours of contact with the cells by the method described in PREPARATION 3 in comparison with the controls (20% fetal calf serum and 20% fetal calf serum + H2O2) by the method described in PREPARATION 3. SR 26276 and the compounds Ro5-4864 significantly inhibit

  significant the pro-apoptotic effect of H202.

Claims (6)

  1. Use of a peripheral benzodiazepine receptor ligand for the   preparation of pharmaceutical compositions capable of inhibiting apoptosis.   2. Use according to claim 1 characterized in that said ligand is a   peripheral benzodiazepine receptor ligand.   3. Use according to claim 1, characterized in that the pharmaceutical compositions are indicated for the treatment of infections by antivirus such as HIV and HTLV 1 and 2 (from the English human immunodeficiency virus and human T lymphocyte virus) and their consequences such as leukemias, myelopathies and arthropathies, hepatitis (C, A, B, F), AIDS, immune deficiencies, cell aging, phenomena of tissue degeneration, inflammation, cell proliferation, diseases infectious, transplant rejection, acute or chronic rheumatoid arthritis, ulcerative colitis, thrombocytopenic purpura, autoimmune hemolytic anemia, type I (insulin-dependent) diabetes, myelodysplastic syndrome, prions, respiratory distress syndrome, prostatic hypertrophy, asthma, atherosclerosis and its thromboembolic complications, kidney and glomerulonephritis, diseases related to abnormal apoptotic activity, eye conditions such as cataracts or glaucoma, osteoporosis, bone fractures, epidermal lesions, restenosis, conditions related to improper proliferation or migration of cells smooth muscles, inflammations of the respiratory system, asbestosis, silicosis, lupus erythematosus, granulomatosis, eosinophilic granulomatosis, gastric and duodenal ulcers, esophagitis, enteritis, gastritis, septicemia, dysfunctions of hematopoiesis and / or lymphopoiesis, cystic fibrosis, psoriasis, peripheral neuropathies, brain damage caused by an attack, diseases causing demyelination in general such as multiple sclerosis, diseases that involve   neuronal degeneration such as vascular dementia, post-   apoplectics, post-traumatic brain injury syndromes, disorders of cerebral anoxia, Alzheimer's disease, senile dementia, subcortical dementia, such as Huntington's chorea and Parkinson's disease, dementia caused by AIDS, neuropathies derived of morbidity or damage to the sympathetic or sensory nerves, and brain diseases, such as cerebral edema, and spinocerebellar degeneration, amyotrophic lateral sclerosis, or diseases requiring cardioprotection such as myocardial infarction, cardiac ischemia, heart failure, coronary vasospasm, angina, heart disease and heart valve disease, inflammatory heart disease, side effects from cardiotoxic drugs or   still in patients undergoing cardiac surgery.   4. Use according to claim 3, characterized in that the pharmaceutical compositions are indicated for the treatment of pathologies linked to an activity abnormal apoptotic.   5. Use according to claim 4, characterized in that the pharmaceutical compositions are indicated for the treatment of a disease chosen from cancer and its metastases; antivirus infections such as HIV and HITV 1 and 2 and their consequences such as leukemia; myelopathies and arthropathies; hepatitis (C, A, B, F); AIDS; immune deficiencies; cell aging; tissue degeneration phenomena; Inflammation; cell proliferation; infectious diseases; rejection of transplants; acute or chronic rheumatoid arthritis; ulcerative colitis; thrombocytopenic purpura; Autoimmune hemolytic anemia; type I (insulin-dependent) diabetes; myelodysplastic syndrome; Huntington's disease; prion diseases; Respiratory distress syndrome;   prostatic hypertrophy; Asthma; Atherosclerosis and its thrombo-complications   embolics; kidney disease and glomerulonephritis.   6. Use according to claim 5, characterized in that the pharmaceutical compositions are indicated for the treatment of transplant rejection or   acute or chronic rheumatoid arthritis.