WO2000004032A1 - Unsaturated phosponates derived from indole - Google Patents

Abstract

The invention concerns compounds corresponding to the general formula (1) wherein: n represents an integer equal to 1, 2 or 3; R represents a hydrogen, R', COR' or SO2R'; R1 represents a hydrogen, an alkyl chain, an acyloxyalkyl, acylthioethyl, a phenyl residue; R2 represents a hydrogen, an alkyl, phenyl, benzyl, or phenethyl radical, as well as their pharmaceutically acceptable salts and solvates. The invention also concerns the methods for preparing said compounds, the pharmaceutical compositions containing them and their use as medicines.

Description

 “Indole derived unsaturated phosphonates”

The present invention relates to unsaturated phosphonates derived from indole, their manufacturing process, the pharmaceutical compositions containing them and their use as medicaments.

Ras oncogenes are the most frequently encountered oncogenes in human tumors and probably play an important role in the proliferation of these tumors. In particular, it has been shown that mutated Ras oncogenes are present in 50% of colon tumors and in more than 90% of pancreatic cancers (Barbacid, M. Ann Rev Biochem 1987; 56: 779-827; Bos, JL Cancer Res 1989; 49: 4682-4689). The transformed Ras proteins, products of these oncogenes, are therefore involved in the proliferation and differentiation processes of cancer cells. Consequently, blocking the function of the Ras proteins must result in the inhibition of the growth of tumor cells dependent on the activation of Ras and / or those which express mutated Ras proteins (Perriπ, D .; Halazy, S. and Hill , BT J Enzyme Inhi 1996; 11: 77-95. Levy, R. Presse Med 1995; 24: 725-729. Sebolt-Leopold, J., S. Emerging Drugs 1996; 1: 219-239. Hamilton, AD and Sebti, SM Drug News Perspect 1995; 8: 138-145; Der, CJ; Cox, AD; Sebti, SM and Hamilton, AD Anti-Cancer Drugs 1996; 7: 165-172 and Halazy, S. and Coll. Drugs of the Future, 1997, 22: 1133-1146).

To be active, the Ras proteins must be associated with the cell membrane, according to a process which depends on post-transcriptional modifications of the Ras gene product, notably involving the addition of a famesyl group on a cysteine residue of the C-terminal tetrapeptide. of the Ras protein. Famesylation of the Ras proteins in which farnesylpyrophosphate acts as a co-substrate is catalyzed by the protein Protein-Faresyl-Transferase (PFTase). The inhibition of PFTase and therefore of the famesylation of the Ras protein makes it possible to control the proliferation of mutated Ras cancer cells. This has been demonstrated using PFTase inhibitors such as BZA-5B (James, GL; Goldstein, JL; Brown, MS et al. Science 1993; 260 [51 16d: 1937-1942) or the derivative L -731, 734 (Kohi, NE; Mosser, SD; DeSolms, SJ et al Science 1993; 260 [5116]: 1934-1937) at the level of cell proliferation but also more recently at the level of tumors grafted in mice (Kohi , NE; Wilson, FR; Mosser, SD et al. Proc Natl Acad Sci USA 1994; 91: 9141-9145. Kohi, NE; Orner, CA; Conner, MW et al. Nature Med 1995; 1: 792-797) . The famesylation of proteins catalyzed by the enzyme PFTase is not limited to the Ras proteins and plays an important role in the function of other proteins also involved in cell proliferation such as for example the Rho B protein or the tyrosine protein. PTPcaax phosphatase (CA Cates et al., Cancer Lett 1997; 110: 49-55). Consequently, PFTase inhibitors find their utility as anticancer agents, capable of controlling cell proliferation in tumors in which protein famesylation plays a determining role.

Furthermore, it has also been shown that the inhibition of geranylgeranylation of certain Ras proteins, a process which is catalyzed by the enzyme geranylgeranyltransferase I (GG Tase I), could also result in the control of the function of the Ras proteins and therefore in controlling their influence on cell differentiation and proliferation (Lerner, EC; Hamilton, AD and Sebti, SM Anti- Cancer D g Design 1997; 12: 229-238; Lerner, EC; Qian, Y .; Hamilton, AD and Sebti, SM J Biol Chem 1995; 270: 26770-26773; Yokoyama, K .; Zimmerman, K .; Scholten, J. and Gelb, MH J Biol Chem 1997; 272: 3944-3952; Osman, H .; Mazet, JL; Maume, G. and Maume, BF Biophys Res Commun 1997; 231: 789-792; Zhang, FL; Kirschmeier, P .; Carr, D. et al. J Biol Chem 1997; 272: 10232-10239). Consequently, compounds capable of inhibiting geranylgeranyltransferase also find their utility in controlling cell proliferation and therefore as anticancer agents.

Controlling the prenylation (famesylation and / or geranylgeranylation) of Ras proteins by PFTase or GGTase I inhibitors also finds utility in the treatment of viral infections such as hepatitis delta virus (δ) infections ( JC Otto and PJ Casey, J Biol Chem 1996; 271: 4569-4572). Furthermore, the role played by Ras proteins in the process of cell proliferation of smooth muscle cells following a vascular lesion (Indolfi et al., Nature Med 1995; 1 (6): 541-545) suggests that the Ras protein prenylation inhibitors are potentially useful for the treatment or prevention of atherosclerosis and restenosis.

The subject of the present invention is a new class of protein prenylation inhibitors, and more particularly inhibitors of the protein Protein-Faresyl-Transferase enzyme (PFTase) and the protein Protein Geranyl Geranyl Transferase (GGTase). distinguish from all the closest derivatives of the prior art by a different chemical structure. Their originality is essentially due to the simultaneous presence of a tryptophan residue, an unsaturated chain and a phosphonate residue which, unexpectedly, confers on these molecules remarkable biological properties, such as for example the potential to selectively inhibit the prenylation of proteins, such as Ras proteins, at the enzymatic level but also at the cellular level. The present invention relates to unsaturated phosphonates derived from indole which are unambiguously distinguished from inhibitors of prenylation of proteins of the prior art by their original chemical structure and by their biological profile. The prior art in this field is illustrated in particular by:

US patent application 329587 and publication J. Biol.

Chem. 268, 7617 (1993) which describe hydroxyfarnesylphosphonic acid as a PFTase inhibitor.

Patent applications EP 534546, US 5523430, US

5510510, WO 9419357, EP 570706 which claim phosphonates derived from famesyl as inhibitors of

PFTase and / or squalene synthase.

Patent application WO 9719091 which claims phosphonates derived from geranylgeranyl as GGTase inhibitors.

Patent application F 9415237 (WO9619484) which claims phosphonates derived from carboxamides.

Patent application EP 618224 which claims phosphonates derived from dipeptides as inhibitors

"Endothelin converting enzyme".

The present invention relates to compounds of general formula (I)

dans laquelle, n représente un nombre entier égal à 1 , 2 ou 3;

in which, n represents an integer equal to 1, 2 or 3;

R represents a hydrogen, R ', COR' or SO ₂ R ', in which R' represents a linear or branched alkyl residue comprising from 1 to 6 carbon atoms, limits included, phenyl, benzyl or phenethyl;

R represents a hydrogen, a linear or branched alkyl chain comprising from 1 to 6 carbon atoms inclusive of limits, an acyloxyalkyl (-CHR OCOR ₅ ), acylthioethyl (-CH ₂ CH ₂ SCOR ₅ ) residue or a phenyl optionally substituted by a or several alkyl (R ₅ ) or alkoxy (OR5) residues in which R ₄ and R ₅ , which are identical or different, represent a linear or branched alkyl residue comprising from 1 to 6 terminal carbon atoms inclusive, or alternatively, R ₄ represents hydrogen.

R ₂ represents a hydrogen, a linear or branched, saturated or unsaturated alkyl residue comprising from 1 to 15 carbon atoms, limits included, a phenyl, a benzyl, or a phenethyl, which may be optionally substituted by one or more residues chosen from OR ", SiMe ₃ , SR", CN, CF ₃ , in which R "represents a linear or branched alkyl residue comprising from 1 to 6 carbon atoms, limits included, phenyl, benzyl or phenethyl.

The compounds of general formula (I) containing 2 asymmetric centers have isomeric forms. The racemates, enantiomers and then diastereoisomers of these compounds and their mixtures in all proportions are also part of the present invention. In particular, the present invention comprises derivatives of general formula (I) in which the asymmetric carbon atom carrying the substituent CO ₂ R ₂ is of configuration (R).

The present invention also includes the compounds of general formula (I) in which the asymmetric carbon atom carrying the substituent CO R is of configuration (S).

The invention also includes the salts, solvates (for example hydrates) of these compounds acceptable for pharmaceutical use.

Among the compounds of general formula (I) forming part of the present invention, a particularly preferred class of compounds corresponds to the compounds of general formula (I) in which n = 2.

Another class of compounds forming part of the present invention which is also particularly appreciated corresponds to the compounds of general formula (I) in which R 1 and R ₂ simultaneously represent hydrogen.

A third class of compounds forming part of the present invention and more particularly appreciated corresponds to the compounds of general formula (I) in which R represents CH ₂ OCO ¹ Bu OR CH ₂ CH ₂ SCO ¹ Bu.

A fourth class of compounds forming part of the present invention and more particularly appreciated corresponds to the compounds of general formula (I) in which R ₂ represents a methyl or isopropyl radical.

Finally, a fifth class of compounds forming part of the present invention and more particularly appreciated corresponds to the compounds of general formula (I) in which R is hydrogen. The present invention also relates to the general process for preparing the derivatives of general formula (I) and more particularly the derivatives of general formula (I) in which R1 and R2 are different from a hydrogen which comprises the formation of an amide bond by reaction (condensation) between a carboxylic acid or a carboxylic acid derivative of general formula (II),

(II) in which n is defined as in the general formula (I), R represents a linear or branched alkyl residue comprising from 1 to 6 terminal carbon atoms inclusive and L represents OH, Cl, imidazole, or the group " COL "represents a carboxylic acid or the activated form of a carboxylic acid suitable for the formation of an amide after reaction with an amino acid derivative (tryptophan derivative) of general formula (III)

in which R is defined as in the general formula (I) and R ₂ is different from hydrogen. Condensation of a carboxylic acid or one of its derivatives of general formula (II) (the preparation of which is described in patent WO 9619484) with an amino acid derivative (an amine) of general formula (III) can be carried out by methods and techniques well known to those skilled in the art for preparing an amide from an amine and a carboxylic acid.

Thus, a particularly preferred method of preparing compounds of general formula (I) in which R 1 and R ₂ are different from hydrogen comprises the condensation of a carboxylic acid of general formula (II) in which L = OH with an amine of general formula (III) using 1- (3-dimethylaminopropyl) -3-ethyl-carbodiimide (EDC) and 3-hydroxy-1,2,3-benzotriazin-4 (3H) -oπe, in the presence of a tertiary amine such as diisopropylethylamine, in a polar aprotic anhydrous solvent such as dichloromethane, at a temperature between -15 ° C and 40 ° C, or alternatively, by using benzotriazol -1 -yloxy-tris (dimethylamino) phosphoniumhexafluorophosphate in the presence of a tertiary amine (N-methylmorpholine) in a polar solvent (DMF or DMSO) at a temperature between -10 ° and 35 ° C. It is understood that the condensation of intermediates (II) and (III) to prepare the compounds of general formula (I) will be carried out under the appropriate conditions which, inter alia, may involve the use of protective groups which are adequate at the level functional groups of intermediates (II) or (III).

When it is desired to prepare a compound of general formula (I) in which the asymmetric carbon of the amino acid residue of formula (III) is of configuration (R), the condensation of a carboxylic acid of general formula (II) will be carried out , according to the methods and techniques described above, with an amino acid derivative of general formula (III) in which the asymmetric carbon is of configuration (R). It is well understood, that, in the same way, a compound of general formula (I) in which this same asymmetric carbon is of configuration (S) will be prepared, by the same methods starting from an amino acid derivative of general formula ( III) in which the asymmetric carbon is of configuration (S).

The derivatives of general formula (I) in which n, R and R, are defined as in the general formula (I) and R ₂ represents a hydrogen are obtained by hydrolysis of a carboxylic ester of general formula (I) in which n , R, Ri and R ₂ are defined as in the general formula (I), by methods well known to those skilled in the art for preparing a carboxylic acid from an ester such as for example the use of 'A sodium, potassium or lithium hydroxide in a polar solvent such as tetrahydrofuran in the presence of water at a temperature between 0 ° C and 40 ° C.

The derivatives of general formula (I) in which n, R and R ₂ are defined as in the general formula (I) and R, represents a hydrogen are prepared from derivatives of general formula (I) in which n, R and R ₂ are defined as in the general formula (I) and Ri represents an alkyl residue comprising from 1 to 6 carbon atoms (preferably an ethyl residue) by reaction with bromotrimethylsilane, in a polar solvent such as dichloromethane, 1 , 2-dichloroethane, acetonitrile, at a temperature between -10 ° C and 60 ° C, in the optional presence of a base such as an amine, for example diisopropylethylamine, N-methylmorpholine, 2,4 , 6-collidine or pyridine.

A process for the preparation of the compounds according to formula I in which

R1 and R2 represent a hydrogen comprises the treatment of a derivative of general formula (I) in which R1 represents an ethyl and R2 represents a methyl or an ethyl with successively the bromotrimethylsilane and a hydroxide of sodium, lithium or potassium.

A process for preparing the compounds according to formula I in which R1 represents CHR ₄ OCOR ₅ or CH ₂ CH ₂ SCOR ₅ comprises the condensation of a compound of general formula (I) in which R1 represents H with XCHR ₄ OCOR ₅ or XCH ₂ CH ₂ SCOR ₅ in which X represents a leaving group such as Cl, Br, I, O-tosyle or O-mesyl.

The compounds of general formula (I) in which R represents CHR ₄ OCOR5 while n, R are defined as in the general formula (I) and R ₂ is different from hydrogen are obtained from the corresponding compounds of general formula ( I) in which Ri represents H by reaction with XCHR4OCOR5 (in which X represents a leaving group such as CI, Br, I, O-tosyl, O-mesyl) in the presence of a base such as a tertiary amine, for example DBU, of a catalyst for example tetrabutylammonium of hydrogen sulphate, optionally of an activating salt for example Nal or Kl in a mixture of polar solvents such as THF, acetonitrile or DMF at a temperature between 25 ° C and 150 ° C.

The compounds of general formula (I) in which R represents CH ₂ CH ₂ SCOR ₅ while n and R are defined as in general formula (I) and R ₂ is different from hydrogen are obtained from the corresponding compounds of general formula (I) in which R- _\ represents H by reaction with HOCH ₂ CH ₂ SCOR ₅ in the presence of 2,4,6-triisopropylbenzene sulfonyl chloride, of an amino base such as pyridine at a temperature between 0 ° C and 100 ° C. An alternative method consists in treating a phosphonic acid of general formula (I) in which R ^ represents H with X CH CH ₂ SCORs in which X is defined as previously by the methods and techniques described above for the preparation of products of formula (I) Ri = CHR ₄ OCORs. The compounds of general formula (I) in which R _T represents a phenyl residue optionally substituted by one or more alkyl or alkoxy residues are prepared by reaction of a derivative of general formula (I) in which R, = H with successively either SOCI ₂ or oxallyl chloride in the presence of DMF, either PCI ₅ or OPCI ₃ in the optional presence of an amino base (for example pyridine, 2,4,6-collidine or triethylamine) in a polar solvent such as THF or dichloromethane at a temperature between -80 ° C and 60 ° C then a phenol optionally substituted by one or more alkyl or alkoxy residues in the presence of an amine such as toludine, DIPEA, 2,4,6 - collidine or DBU in a polar solvent such as DMF, THF.

When it is desired to isolate a compound of general formula (I) in which Ri and / or R ₂ represents a hydrogen, in the form of a salt, for example salt by addition with a base, this can be achieved by treating the free acid of general formula (I) wherein R and / or R ₂ represents hydrogen with a suitable base, preferably in an equivalent amount.

Such salts acceptable for pharmaceutical use include the cation salts such as lithium, sodium, potassium, calcium, magnesium, aluminum, ammonium, quaternary ammoniums and the salts formed after reaction with an amine non-toxic organic, such as for example lysine.

The compounds according to the present invention can exist in the form of racemates, enantiomers, diastereoisomers and their mixtures in all proportions. When the processes described above for preparing the compounds of the invention give mixtures of diastereoisomers, these isomers can be separated by conventional methods such as preparative chromatography.

When the new compounds of general formula (I) have one or more asymmetric centers, they can be prepared in the form of a racemic mixture or in the form of enantiomers, either by enantioselective synthesis or by resolution.

The following examples illustrate the invention without, however, limiting its scope.

EXAMPLE 1

(2S) 3- (1 / V-indol-3-yl) -2 - [(4E, 8E) 2- (dihydroxyphosphoryl) - 5,9,13-trimethyltetradeca-4,8,12-trienoylamino] acid propionic)

The preparation of compound Λ is carried out as follows: Preparation of compound 1A: Le (2S) 3- (1H-indol-3-yi) -2 - [(4E, 8E) 2- (diethoxyρhosp oryl) -5.9, 13-trimethyltetradeca-4,8 ₎ 12-trienoylamino] methyl propionate

(4E.8E) 2- (diethoxyphosphoryl) -5,9,13-trimethyltetradeca-4,8,12-trienoic acid (patent WO 9619484 A) (4.14 g; 10.3 mmol) is dissolved in a mixture of ethyl acetate (100 ml) and chloroform (100 ml) at room temperature and under a nitrogen atmosphere. 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI; 2.18 g; 11.4 mmol) and 3-hydroxy-1,2,3-benzotriazin-4 (3H) -one (HOOBT; 1.86 g; 11.4 mmol) are added. The reaction mixture is stirred for 30 minutes then the hydrochloride of L-tryptophan methylester (3.16 g; 12.4 mmol) and diisopropylethylamine (5.4 ml; 31 mmol) are introduced. The reaction medium is stirred for 15 h at 20 ° C. then diluted in ethyl acetate (300 ml) and water (300 ml). The organic phase is washed once with 200 ml of a saturated aqueous NaCl solution and washed once with 200 ml of a saturated aqueous NH4CI solution. It is then dried over magnesium sulfate, filtered and concentrated. The crude product obtained is purified by flash chromatography with a gradient (0 to 2%) of methanol in dichloromethane.

Mass obtained: 4.92 g (79%)

¹ H-NMR (400 MHz; DMSO-dβ), mixture of two diastereoisomers. δ: 10.91 and 10.87 (s, 1H, NH); 8.55-8.40 (m, 1H, NH); 7.46 (d, 1 H, 7.7 Hz); 7.35-7.30 (m, 1H); 7.24 and 7.12. (brs, 1 H); 7.09-7.00 (m, 1H); 7.00-6.95 (m, 1H); 5.12-4.90 (m, 3H); 4.59 and 4.52 (q, 1H, 6.4Hz); 4.10-3.75 (m, 4H); 3.54 (s, 3H); 3.15-2.90 (m, 3H); 2.60-2.40 (m, 1H); 2.40-2.20 (m, 1H); 2.10-1.75 (m, 8H); 1.65-1.45 (m, 12H); 1.25-1.00 (m, 6H).

Preparation of compound 1 B: (2S) 3- (1H-indol-3-yl) -2 - [(4E, 8E) 2- (diet oxyphosphoryl) -5,9,13-trimethyltetradeca-4,8 acid , 12- triénoylamino] propionique

The derivative 1A (943 mg; 1.57 mmol) is treated with lithium hydroxide (1 M in water; 1.57 ml; 1.57 mmol) in tetrahydrofuran (2 ml) at room temperature. The solution is stirred for 1 hour then poured into ethyl acetate (30 ml) and brought back to pH = 3 by addition of hydrochloric acid (1 N in water). The organic phase is separated, dried over magnesium sulfate and concentrated. The 1JB product is purified by flash chromatography with a gradient (1 to 5%) of methanol in dichloromethane.

Mass obtained: 695 mg (75%)

^ H-NMR (400 MHz; DMSO-dβ), mixture of two diastereoisomers. δ:

12.27 (m, 1H, OH); 10.85 and 10.80 (s, 1H, NH); 8.18 (m, 1H, NH); 7.55-

7.48 (m, 1H); 7.35-7.27 (m, 1H); 7.23 and 7.09 (brs, 1H); 7.05-6.98 (m, 1H); 6.98-6.90 (m, 1H); 5.10-4.90 (m, 3H); 4.55-4.35 (m, 1H); 4.10-3.75 (m, 4H); 3.25-2.90 (m, 3H); 2.55-2.35 (m, 1H); 2.35-2.15 (m, 1H); 2.10-1.75, (m, 8H); 1.70-1.45 (m, 12H); 1.30-1.00 (m, 6H).

1- Diester 1B (590 mg; 1.0 mmol) is placed in solution in dry dichloromethane (15 ml). 2,4,6-collidine (665 μl; 5.0 mmol) and then bromotrimethylsilane (TMSBr; 665 μl; 5.0 mmol) are added at 0 ° C. The reaction is brought slowly to room temperature and then stirred for 20 hours. The reaction mixture is co-evaporated twice with toluene. The crude product is taken up in ethyl acetate (50 ml) and acidified with an aqueous solution of hydrochloric acid (1 N) to pH approximately equal to 2. The organic phase is dried over magnesium sulfate, filtered and concentrated. The residual oil is dissolved in dichloromethane and filtered through glass wool. The filtrate is concentrated to give derivative 1 in the form of a hard, clear and slightly colored oil.

Mass obtained: 473 mg (88%)

¹ H-NMR (400 MHz; DMSO-dβ), mixture of two diastereoisomers. δ: 12.5-11.0 (brs, 1H, OH); 10.72 (s, 1H, NH); 8.01 (brs, 1H, NH); 7.50 (brd, 1H); 7.29 (brd, 1H); 7.25 and 7.12 (brs, 1H); 7.03 (brt, 1H); 6.94 (brt, 1H); 5.15-4.95 (m, 3H); 4.60-4.40 (m, 1H); 3.25-2.95 (m, 2H); 2.80-2.55 (m, 1H); 2.55-2.20 (m, 2H); 2.10-1.75 (m, 8H); 1.70-1.45 (m, 12H).

The derivative is dissolved in methanol (3 ml) then an aqueous solution (50%) of lysine is added (294 μl, 1.25 eq). Salt precipitates. It is filtered, rinsed with a minimum of a petroleum ether - methanol mixture (50 - 50).

Mass obtained: 470 mg (white solid)

Elemental analysis for: C28H39N2O6P; 1.8 C6H1 N2O2; 0.8 H2O Calculated C: 57.67; H: 8.21; N: 9.71 Found C: 57.65; H: 8.29; N: 9.65.

Mass (ESI-) 429 (MH) ⁺ ; 551 (M-2H + Na) ⁺ .

EXAMPLE

(2R, 2S) 3- (1tf-indol-3-yl) -2 - [(4E, 8E) 2- (dihydroxyp osphoryl) - 5,9,13-trimethyltetradeca-4,8,12-trienoylamino acid ] propionic {V)

V

The derivative Y is prepared from D, L tryptophan according to the same procedures used in the synthesis of 1. Mass obtained: 212 mg H-NMR (400 MHz; DMSO-de), mixture of diastereoisomers. δ: 12.5-

10.5 (brs, 1H, OH); 10.80 (s, 1H, NH); 8.00 (m, 1H, NH); 7.50 (brd, 1H); 7.29 (brd, 1H); 7.25 and 7.12 (brs, 1H); 7.03 (brt, 1H); 6.94 (brt, 1H); 5.15-4.95 (m, 3H); 4.60-4.40 (m, 1H); 3.25-2.90 (m, 2H); 2.75-2.60 (m, 1H); 2.55-2.20 (m, 2H); 2.10-1.75 (m, 8H); 1.70-1.45 (m, 12H).

The derivative Y is dissolved in methanol (3 ml), an aqueous solution (50%) of lysine is added (97 μl, 1.0 eq) followed by 50 ml of water. The white opalescent mixture is concentrated under reduced pressure and then lyophilized.

Mass obtained: 200 mg (off-white solid)

Elementary analysis for: C28 ^H 39N2θ6P; 1, 3 C6H1 N2O2; 2.5 H2O

Calculated C: 56.16; H: 8.19; N: 8.41

Found C: 56.20; H: 7.87; N: 8.41.

Mass (ESI +) 431 (M + H) ⁺

EXAMPLE 2

(2S) 3- (1H-indol-3-yl) -2 - [(4E, 8E) 2- (dihydroxyphosphoryl) -5,9,13- trimethyltetradeca-4,8,12-trienoylamino] methyl propionate ( 2)

Compound 2 is obtained by hydrolysis of derivative 1A according to the procedure used for the preparation of from the following reagents: 1A (3.85 g; 6.4 mmol); TMSBr (4.2 ml; 32 mmol); 2,4,6-collidine (4.2 ml; 32 mmol) in dichloromethane (90 ml).

Mass obtained: 3.90 g (98%, oil) 1 H-NMR (400 MHz; DMSO-dβ), mixture of two diastereoisomers. δ:

10.91 and 10.87 (s, 1H, NH); 8, 10-8.40 (m, 1H, NH); 7.50-7.42 (m, 1H); 7.35-7.30 (m, 1H); 7.24 and 7.13 (brs, 1H); 7.05 (brt, 1H); 7.00-6.95 (m, 1H); 5.12-4.90 (m, 3H); 4.57 and 4.51 (brq, 1H); 3.51 and 3.49 (s, 3H); 3.15-3.00 (m, 2H); 2.80-2.60 (m, 1H); 2.60-2.40 (m, 1H); 2.40-2.20 (m, 1H); 2.10-1.75 (m, 8H); 1.65-1.45 (m, 12H).

EXAMPLE 3

(2S) 3- (1W-indol-3-yl) -2 - {(4E, 8E) 2- [bis (2,2- dimethylpropionyloxymethoxy) phosphoryl] -5,9,13-trimethyltetradeca-

4,8,12-triénoylamino} methyl propionate (3)

Derivative 2 (2.96 g; 5.43 mmol) is dissolved in THF (23 ml) and acetonitrile (23 ml) in the presence of tetrabutylammonium hydrogen sulfate (930 mg; 5.43 mmol) of DBU (810 μl; 5.43 mmol) of pivaoyl chloride (3.15 ml; 21.7 mmol) and sodium iodide (815 mg; 5.43 mmol) under a nitrogen atmosphere and at room temperature. The reaction mixture is then heated to 60 ° C. At the third hour of heating, peony chloride (3.15 ml; 21.7 mmol) and DBU (810 μl; 5.43 mmol) are added. At the seventh hour of heating, peony chloride (2 ml; 14 mmol) is added. The reaction mixture is heated a total of 22 hours. It is then cooled and concentrated. Derivative 3 is purified by flash chromatography with a gradient (0 to 2%) of methanol in dichloromethane.

Mass obtained: 1.54 g (36%, hard oil)

¹ H-NMR (400 MHz; DMSO-de), mixture of two diastereoisomers. δ:

10.89 and 10.88 (s, 1H, NH); 8.60 (d, 1H, NH, 7.4 Hz); 7.50-7.40 (m, 1H); 7.35-7.30 (m, 1H); 7.20 and 7.10 (brs, 1H); 7.05 (brt, 1H); 7.00-6.93 (m, 1H); 5.65-5.45 (m, 4H); 5.10-4.90 (m, 3H); 4.60-4.50 (m, 1H); 3.55 and 3.51 (s, 3H); 3.25-3.0 (m, 3H); 2.55-2.35 (m, 1H); 2.35-2.20 (m, 1H); 2.05-1.75 (m, 8H); 1.65-1.45 (m, 12H); 1.17 (s, 18H). Rf = 0.35 and 0.39 (5% MeOH in CH ₂ CI ₂ )

Mass obtained: 227 mg (5%, hard oil) ¹ H-NMR (400 MHz; DMSO-dβ), least polar diastereoisomer, δ:

10.89 (s, 1H, NH); 8.60 (d, 1H, NH, 7.4 Hz); 7.47 (d, 1H, 7.8Hz); 7.32 (d, 1H, 8.0 Hz); 7.20 (d, 1H, 1.9Hz); 7.06 (t, 1H, 7.5Hz); 6.97 (t, 1H, 7.4Hz); 5.65-5.45 (m, 4H); 5.10-5.00 (m, 3H); 4.55 (q, 1H, 7.1 Hz); 3.51 (s, 3H); 3.25-3.0 (m, 3H); 2.55-2.35 (m, 1H); 2.35-2.20 (m, 1H); 2.05-1.75 (m, 8H); 1.65-1.45 (m, 12H); 1.17 (s, 18H). Rf = 0.39 (5% MeOH in CH ₂ CI ₂ ) EXAMPLE 4

(2S) 3- (1H-indol-3-yl) -2 - ((4E, 8E) 2- {bis [2- (2,2- dimethylpropionylsulfanyl) ethoxy] phosphoryl} -5,9,13- trimethyltetradeca -4,8,12-trienoylamino) methyl propionate (4)

Derivative 2 (841 mg; 1.54 mmol) is dissolved in pyridine (20 ml) in the presence of 2- (2,2-dimethylpropionylsulfanyl) ethanol (1.25 g; 7.72 mmol). The solution is concentrated under reduced pressure, then again dissolved in pyridine (20 ml) and concentrated. The oily residue is placed under a nitrogen atmosphere and dissolved in pyridine (12 ml). 2,4,6-Triisopropylbenzene sulfonyl chloride (2.41 g, 7.72 mmol) is added and the reaction is stirred for 23 hours at room temperature. The reaction mixture is then diluted in 180 ml of dichloromethane, washed once with 160 ml of a saturated aqueous sodium bicarbonate solution, once with 120 ml of water, then dried over magnesium sulfate and concentrated. The resulting solution is co-evaporated twice toluene. Derivative 4 is purified by flash chromatography with a gradient (0 to 1%) of methanol in dichloromethane.

Mass obtained: 234 mg (18%, hard oil) ¹ H-NMR (400 MHz iDMSO-dβ), mixture of two diastereoisomers. δ 10.90 and 10.88 (s, 1H, NH); 8.53 (d, 1H, NH, 7.4 Hz); 7.50-7.40 (m, 1H) 7.32 (brd, 1H); 7.23 and 7.11 (brs, 1H); 7.06 (brt, 1H); 7.00-6.93 (m, 1H) 5.15-4.90 (m, 3H); 4.59 and 4.53 (q, 1H, 6.7Hz); 4.15-3.75 (m, 4H); 3.54 and 3.52 (s, 3H); 3.20-2.95 (m, 7H); 2.55-2.35 (m, 1H); 2.35-2.20 (m, 1H); 2.10-1.70 (m, 8H); 1.70-1.45 (m, 12H); 1, 20-1, 10 (brs, 18H). Rf = 0.15 (5% MeOH in CH ₂ CI ₂ )

The derivatives of the present invention are inhibitors of prenylation of proteins and more particularly of famesylation of Ras proteins as shown by studies of inhibition of famesyl transferase protein and of geranylgeranyl transferase protein.

A) Evaluation of the inhibition of the Farnesyl Transferase Protein:

Principle

The famesylation of the dansylated peptide GCVLS, catalyzed by the protein farnesyl transferase enzyme, causes a change in the emission spectrum of the dansyl group, and in particular an increase in the emission at 505 nm when the molecule is excited at 340 nm. Measured with a spectrofluorometer, this emission is proportional to the activity of the enzyme (Pompliano et al., J. Am. Chem. Soc. 1992; 114: 7945-7946).

Equipment

Reaction buffer: 55 mM TRIS / HCI Ph 7.5; 5.5 mM DTT; 5.5 mM MgCl ₂ ; 110 μM ZnCI ₂ , 0.22% B-octyl-B D-glucopyrannoside. Substrates:

Faresylpyrophosphate (FPP), (Sigma)

Dansylated peptide dansyl-GCVLS (Neosystem / Strasbourg, France) Enzyme: Farnesyl protein transferase is partially purified from beef brain by ion-exchange chromatography on Q-Sepharose (Pharmacia) (Moores et al., J. Biol. Chem. 1991, 266: 14603-14610, Reiss et al., Cell 1990, 62: 81-88).

Method

The reaction mixture containing 2.2 μM of FPP, 2.2 μM of dansyl GCVLS with or without (zero) the quantity of enzyme giving an intensity of 100 to the spectrofluorimeter after incubation for 10 minutes at 37 ° C., is prepared on ice. In an Eppendorf tube, 360 μL of reaction mixture are mixed with 40 μl of test product 10 times concentrated or of solvent, and incubated for 10 minutes at 37 ° C. The reaction is stopped on ice and the intensity of the fluorescence is measured (excitation 340 nm, slit 4 nm, emission 505 nm, slit10 nm). The tests are carried out in duplicate. The results are expressed as a percentage of inhibition. Under these conditions, the derivatives of the present invention have been identified as potent inhibitors of farnesyl protein transferase (IC ₅₀ <10 μM).

B) Evaluation of the inhibition of Geranylqeranyl Protein Transferase I

Equipment

Reaction buffer:

55 mM TRIS / HCI Ph 7.5; 5.5 mM DTT; 5.5 mM MgCl ₂ ; 110 μM ZnCI ₂ , 0.22% N-octyl-B D-glucopyrannoside. Substrates:

³ H-geranylgeranyl pyrophosphate (GGPP), 66 μM, 15 Cl / mmol, (Isotopchim)

Recombinant Rho-GST Protein Enzyme:

GGPT I is partially purified from beef brain by ion exchange chromatography on Q-sepharose (Pharmacia); elution at 0.23 and 0.4 M NaCI resp.

(Moores et al., J. Biol. Chem. 1991, 266: 14603-14610; Reiss et al., Cell 1990, 62: 81-88).

Method

The reaction mixture containing 220 nM 3H-GGPP; 0.5 μM of rho- GST with or without (zero) 5 μl of GGPT 1 / test, is prepared on ice. In an Eppendorf tube, 45 μl of reaction mixture are mixed with 5 μl of test product 10 times concentrated or of solvent, and incubated 45 min at 37 ° C. A 45 μl aliquot is placed on a numbered P81 phosphocellulose filter, washed in a 1/1 (V / V) mixture of 95% ethanol and 75 mM phosphoric acid and counted by scintillation. The tests are carried out in duplicate.

The results are expressed as a percentage of inhibition.

The derivatives of the present invention are inhibitors of the enzymes which catalyze the prenylation of proteins and more particularly of the enzymes PFTase and GGTase. They are distinguished from the closest derivatives of the prior art, not only by their original chemical structure but also by their biological activity and more particularly by their effectiveness in inhibiting prenylation enzymes such as PFTase. Thus, by way of illustration, the derivative of the present invention has proved, quite unexpectedly, a much better PFTase inhibitor and much more selective than the derivatives closest to the art. previous as indicated by the results collated in the following table.

Also to be considered as forming part of the present invention are pharmaceutical compositions containing, as active ingredients, a compound of general formula (I) or a physiologically acceptable salt of a compound of formula (I) combined with one or more agents therapeutic, such as for example anticancer agents such as for example cytotoxic anticancer agents such as navelbine, vinflunine, taxol, taxotere, 5-fluorouracil, methotrexate, doxorubicin, camptothecin, etoposide, cis- platinum or BCNU.

The present invention also relates to pharmaceutical compositions containing as active ingredient a compound of general formula (I) or one of its salts acceptable for pharmaceutical use, mixed or combined with a suitable excipient. These compositions can take, for example, the form of solid, liquid compositions, emulsions, lotions or creams.

The present invention also relates to the compounds of general formula I for their application as active therapeutic substances for the treatment or prevention of cancers, both curative and preventive treatment of disorders related to famesylation and to geranylgeranylation of proteins, the treatment or the prevention of restenosis, atherosclerosis and viral infections caused by the delta (δ) liver virus.

As solid compositions for oral administration, tablets, pills, powders (gelatin capsules, cachets) or granules can be used. In these compositions, the active principle according to the invention is mixed with one or more inert diluents, such as starch, cellulose, sucrose, lactose or silica, under a stream of argon. These compositions can also comprise substances other than diluents, for example one or more lubricants such as magnesium stearate or talc, a dye, a coating (dragees) or a varnish.

As liquid compositions for oral administration, there may be used pharmaceutically acceptable solutions, suspensions, emulsions, syrups and elixirs containing inert diluents such as water, ethanol, glycerol, vegetable oils or oil paraffin. These compositions can include substances other than diluents, for example wetting, sweetening, thickening, flavoring or stabilizing products.

The sterile compositions for parenteral administration can preferably be aqueous or non-aqueous solutions, suspensions or emulsions. As solvent or vehicle, water, propylene glycol, a polyethylene glycol, vegetable oils, in particular olive oil, injectable organic esters, for example ethyl oleate or other suitable organic solvents, can be used. These compositions can also contain adjuvants, in particular wetting agents, isotonizers, emulsifiers, dispersants and stabilizers. Sterilization can be done in several ways, for example by aseptic filtration, by incorporating sterilizing agents into the composition, by irradiation or by heating. They can also be prepared in the form of sterile solid compositions which can be dissolved at the time of use in sterile water or any other sterile injectable medium.

The compositions for rectal administration are suppositories or rectal capsules which contain, in addition to the active product, excipients such as cocoa butter, semi-synthetic glycerides or polyethylene glycols.

The compositions for topical administration can be, for example, creams, lotions, eye drops, mouthwashes, nasal drops or aerosols.

The doses depend on the desired effect, on the duration of the treatment and on the route of administration used; they are generally between 0.001 g and 1 g (preferably between 0.005 g and 0.25 g) per day, preferably orally for an adult with unit doses ranging from 0.1 mg to 500 mg of active substance, preferably from 1 mg to 50 mg.

In general, the doctor will determine the appropriate dosage based on age, weight and all other factors specific to the subject to be treated. The following examples illustrate compositions according to the invention. In these examples, the term "active component designates one or more (generally one) of the compounds of formula (I) according to the present invention.

Tablets

They can be prepared by direct compression or by passing through wet granulation. The direct compression procedure is preferred, but it may not be suitable in all cases depending on the doses and the physical properties of the active component.

A - By direct compression mg for 1 tablet active component 10.0 microcrystalline cellulose B.P.C. 89.5 magnesium stearate 0.5

100.0

The active component is passed through a sieve with a mesh opening of 250 μm on a side, it is mixed with the excipients and it is compressed using 6.0 mm punches. Tablets with other mechanical strengths can be prepared by varying the compression weight with the use of appropriate punches.

B - Wet granulation mg for one tablet active ingredient 10.0 lactose Codex 74.5 starch Codex 10.0 corn starch pregelatinized Codex 5.0 magnesium stearate 0 3

Compression weight 100.0 The active component is passed through a sieve with a mesh opening of 250 μm and mixed with lactose, starch and pregelatinized starch. The mixed powders are moistened with purified water, they are granulated, dried, sieved and mixed with magnesium stearate. The lubricated granules are put into tablets as for the formulas by direct compression. A coating film can be applied to the tablets using suitable film-forming materials, for example methylcellulose or hydroxypropyl-methyl-cellulose, according to conventional techniques. Sugar tablets can also be coated.

Capsules

mg for one active ingredient capsule 10.0

^* starch 1500 89.5 magnesium stearate Codex 0.5

Filling weight 100.0

^* a form of directly compressible starch from the firm Colorcon Ltd, Orpington, Kent, United Kingdom.

The active component is passed through a sieve with a mesh opening of 250 μm and mixed with the other substances. The mixture is introduced into hard gelatin capsules No. 2 on an appropriate filling machine. Other dosage units can be prepared by changing the filling weight and, if necessary, changing the size of the capsule. Syrup mg per 5 ml dose active ingredient 10.0 sucrose Codex 2750.0 glycerin Codex 500.0 buffer) flavor) color) qs preservative) distilled water 5.0

The active component, the buffer, the flavor, the color and the preservative are dissolved in part of the water and the glycerin is added. The remainder of the water is heated to 80 ° C. and the sucrose is dissolved therein and then cooled. The two solutions are combined, the volume is adjusted and mixed. The syrup obtained is clarified by filtration.

Suppositories

Active ingredient 10.0 mg

"Witepsol H15 supplement to 1.0 g

^* Brand sold for Adeps Solidus of the European Pharmacopoeia. A suspension of the active component in Witepsol H15 is prepared and introduced into a suitable machine with 1 g suppository molds. Liquid for administration by intravenous injection

active ingredient g / 1 water for injection Codex 2.0 complement to 1000.0

Sodium chloride can be added to adjust the tone of the solution and adjust the pH to maximum stability and / or to facilitate the dissolution of the active component by means of a dilute acid or alkali or by adding buffer salts. appropriate. The solution is prepared, clarified and introduced into suitable size vials which are sealed by melting the glass. The liquid for injection can also be sterilized by heating in an autoclave according to one of the acceptable cycles. The solution can also be sterilized by filtration and introduced into a sterile ampoule under aseptic conditions. The solution can be introduced into the ampoules in a gaseous atmosphere.

Cartridges for inhalation g / cartridge active ingredient micronized 1, 0 lactose Codex 39.0

The active component is micronized in a fluid energy mill and made into fine particles before mixing with lactose for tablets in a high energy mixer. The powder mixture is introduced into hard gelatin capsules No. 3 on an appropriate encapsulating machine. The contents of the cartridges are administered using a powder inhaler. Pressure aerosol with metering valve mg / dose for 1 can micronized active component 0.500 120 mg oleic acid Codex 0.050 12 mg trichlorofluoromethane for pharmaceutical use 22.25 5.34 g dichlorodifluoromethane for pharmaceutical use 60.90 14.62 g

The active component is micronized in a fluid energy mill and put into the state of fine particles. The oleic acid is mixed with the trichlorofluoromethane at a temperature of 10-15 ° C. and the micronized drug is introduced into the solution using a mixer with a high shearing effect. The suspension is introduced in measured quantity into aluminum aerosol cans on which are fixed appropriate metering valves delivering a dose of 85 mg of the suspension; dichlorodifluoromethane is introduced into the boxes by injection through the valves.

Claims

1. Compounds corresponding to the general formula (I)

in which, n represents an integer equal to 1, 2 or 3; R represents a hydrogen, R ', COR' or SO R ', in which R' represents a linear or branched alkyl residue comprising from 1 to 6 carbon atoms, limits included, a phenyl, benzyl or phenethyl; R ^ represents a hydrogen, a linear or branched alkyl chain comprising from 1 to 6 carbon atoms limits inclusive, an acyloxyalkyl residue (-CHR OCOR ₅ ), acylthioethyl (-CH ₂ CH ₂ SCOR ₅ ) or a phenyl optionally substituted by one or more alkyl (R ₅ ) or alkoxy (OR5) residues in which R ₄ and R ₅ , which are identical or different, represent a linear or branched alkyl residue comprising from 1 to 6 carbon atoms, limits included, or alternatively, R ₄ represents hydrogen, R ₂ represents a hydrogen, a linear or branched, saturated or unsaturated alkyl residue comprising from 1 to 15 terminal carbon atoms inclusive, a phenyl, a benzyl, or a phenethyl, which may be optionally substituted by one or more residues chosen from OR " , SiMβ ₃ , SR ", CN, CF ₃ , in which R" represents a linear or branched alkyl residue comprising from 1 to 6 terminal carbon atoms inclusive, a phenyl, benzyl or phenethyl, as well as their salts, solvates acceptable for pharmaceutical use, said compounds possibly existing in the form of racemates, enantiomers, diasteroisomers and their mixtures in all proportions. 2. Compounds according to Claim 1, characterized in that n = 2. 3. Compounds according to claim 1 or 2 characterized in that R ^ and R ₂ simultaneously represent hydrogen. 4. Compounds according to claim 1 or 2 characterized in that R ^ represents CH ₂ OCθ'Bu OR CH ₂ CH ₂ SCθ'Bu. 5. Compounds according to one of claims 1, 2 or 4 characterized in that R ₂ represents CH ₃ or CH (CH3) 2- 6. Compounds according to one of claims 1 to 5 characterized in that R is hydrogen. 7. Process for the preparation of compounds according to claim 1 for which the groups R1 and R2 in the general formula (I) are different from hydrogen, a process which comprises a step of condensation of a carboxylic acid or of a derivative of this carboxylic acid of general formula (II)

(he) in which R 1 represents a linear or branched alkyl residue comprising from 1 to 6 carbon atoms, limits included, and the group "COL" represents a carboxylic acid residue or an activated form of this carboxylic acid, with an amino acid derivative of general formula (III) in which R ₂ is different from hydrogen

by techniques and methods well known to those skilled in the art for preparing an amide from a carboxylic acid and an amine. 8. A process for preparing the compounds according to claim 1 for which the groups Ri and R ₂ of the general formula (I) represent a hydrogen, process which comprises the treatment of a derivative of general formula (I) in which R ^ represents an ethyl and R ₂ represents a methyl or an ethyl with successively the bromotrimethylsilane and a sodium, lithium or potassium hydroxide. 9. Process for preparing the compounds according to claim 1 for which the group Ri represents CHR ₄ OCORs or CH ₂ CH ₂ SCOR ₅ which comprises the condensation of a compound of general formula (I) in which Ri represents H with XCHR4OCOR5 or XCH ₂ CH ₂ SCOR ₅ in which X represents a leaving group such as Cl, Br, I, O-tosyl or O-mesyl. 10. Compounds according to one of claims 1 to 6 or compounds obtained by the process according to one of claims 7 to 9 for their application as therapeutically active substances. 11. Compounds according to claim 10 for the treatment or prevention of cancers. 12. Compounds according to claim 10 for both curative and preventive treatment of disorders related to famesylation and to geranylgeranylation of proteins. 13. Compounds according to claim 10 for the treatment or prevention of restenosis and atherosclerosis. 14. Compounds according to claim 10 for the treatment or prevention of viral infections due to the delta hepatic virus (δ). 15. Pharmaceutical compositions containing as active ingredient a compound according to one of claims 1 to 6 or one of its salts acceptable for pharmaceutical use and an appropriate excipient.