Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
  • C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
  • C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
  • C07C323/57—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups
  • C07C323/58—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups with amino groups bound to the carbon skeleton
  • C07C323/59—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups with amino groups bound to the carbon skeleton with acylated amino groups bound to the carbon skeleton
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

• the present invention relates to new famesyl transferase inhibitors of general formula:
• the inhibition of farnesyl transferase and, therefore, of the mesylation of the Ras protein blocks the ability of the mutated Ras protein to transform normal cells into cancer cells.
• the C-terminal sequence of the Ras gene contains the motif "CAAX" or "Cys-
• Aaa ⁇ -Aaa2-Xaa "in which Aaa represents an aliphatic amino acid and Xaa represents any amino acid. It is known that tetrapeptides with a CAAX sequence can inhibit the f mesylation of the Ras protein. For example, in PCT application WO 91 / 16340 and in application EP 0 461 869 have been described peptides inhibitors of the famesyl transferase Cys-Aaai -Aaa2-Xaa which are particularly represented by the peptides
• Cys-Val-Leu-Ser, Cys-Val-Ue-Met and Cys-Val-Val-Met which show their inhibitory activity at concentrations close to 10 ⁇ 6 or 10 "'M.
• Ri represents a radical of general formula YS-Ai - in which Y represents a hydrogen atom, or an amino acid residue or a fatty acid residue or a radical alkyl or alkoxycarbonyl or a radical R4-S- in which R4 represents an alkyl radical containing 1 to 4 carbon atoms optionally substituted by a phenyl radical or a radical of general formula:
• Aj represents a straight or branched alkylene radical containing 1 to 4 carbon atoms optionally substituted at ⁇ of group> C (Xj) (Y ⁇ ) with an amino radical, alkylamino containing 1 to 6 carbon atoms in a straight or branched chain, dialkylamino in which each alkyl part contains 1 to 6 carbon atoms in a straight or branched chain, alkanoylamino containing 1 to 6 carbon atoms in a straight or branched chain or alkoxycarbonylamino the alkyl part of which contains 1 to 6 carbon atoms in a straight or branched chain,
• X represents an oxygen or sulfur atom
• R2 represents an alkyl, alkenyl or straight or branched alkynyl radical containing 1 to 6 carbon atoms optionally substituted by a hydroxy radical, alkoxy containing 1 to 4 carbon atoms, mercapto, alkylthio containing 1 to 4 carbon atoms, alkylsulfinyl containing 1 to 4 carbon atoms or alkylsulfonyl containing 1 to 4 carbon atoms, it being understood that, when R2 represents an alkyl radical substituted by a hydroxy radical, R2 can form with the carboxy radical to a lactone, R'2 represents a hydrogen or a straight or branched alkyl radical containing 1 to 6 carbon atoms, and R represents a hydrogen atom or an alkyl radical containing 1 to 6 carbon atoms optionally substituted by an alkoxy radical containing 1 to 4 carbon atoms, alkylthio containing 1 to 4 carbon atoms, alkylsulfinyl containing 1 to 4
• Ri represents a radical of formula YS-Aj- in which Y represents a hydrogen atom or a lysine residue or a fatty acid residue containing up to 20 carbon atoms and Ai represents an ethylene or propylene radical optionally substituted by a amino radical,
• X represents an oxygen atom
• R2 represents an alkyl radical containing 1 to 4 carbon atoms optionally substituted by a hydroxy, methoxy, mercapto, methylthio, methylsulfinyl or methylsulfonyl radical
• R'2 represents a hydrogen atom or a methyl radical
• R represents a hydrogen atom or an alkyl radical containing 1 to 4 carbon atoms, optionally substituted by an alkoxy radical, or a phenyl radical. More particularly still, 97/03047 PC17FR96 / 01071
• Rj represents a radical of formula Y-S-Ai - in which Y represents a hydrogen atom and A ⁇ represents an ethylene or propylene radical optionally substituted by an amino radical,
• R'i represents a hydrogen atom
• X represents an oxygen atom
• R2 represents a methyl, ethyl, propyl or butyl radical optionally substituted by a hydroxy, methoxy, mercapto or methylthio radical
• R * 2 represents a hydrogen atom
• R represents a hydrogen atom or an alkyl radical containing 1 to 4 carbon atoms.
• R 1 C (X 1 ) (Y 1 ) (NR ' 1 ) and R'2CH (NR' 2 ) CO-OH preferably have the configuration of natural amino acids.
• the present invention also relates to the mineral or organic salts of the products of general formula (I).
• the new products according to the invention can be prepared by applying known methods derived from the methods used more particularly in peptide chemistry for the assembly of chains.
• X represents an oxygen atom are obtained from 3- or 4-nitro- acid naphthalene-1-carboxylic on which an amino acid of general formula is condensed:
• R2 and R'2 are defined as above and R 'represents an alkyl radical containing 1 to 4 carbon atoms, optionally substituted by a phenyl radical, preferably a tert-butyl radical, operating in the presence of an agent condensation, such as hydroxybenzotriazole and dicyclohexylcarbodiimide, and of a base, such as triethylamine, in an organic solvent, such as dimethylformamide, to give a product of general formula:
• a reducing agent such as sodium cyanoborohydride, sodium borohydride, sodium triacetoxyborohydride or hydrogen in presence of a catalyst.
• the reaction is carried out in an organic solvent such as an alcohol such as methanol optionally in combination with another organic solvent such as an ether such as tetrahydrofuran. It is particularly advantageous to operate in an anhydrous medium.
• the protective groups are replaced by hydrogen atoms by application of the usual techniques.
• the Boc or trityl or tert-butyl protecting groups can be replaced by hydrogen atoms by means of trifluoroacetic acid in the presence of ethanedithiol or triethylsilane.
• the products of general formula (I) in which X represents a sulfur atom can be obtained from a product of general formula (IV) in which X represents an oxygen atom by thionation, generally with the reagent of Lawesson, then by performing the reduction, condensation or reductive amination steps as appropriate and deprotection described above for the preparation of a product of general formula (I) in which X represents an oxygen atom.
• the 3-nitro-naphthalene-1-carboxylic acid can be prepared according to the method described by T. NAKAYAMA et al., Chem. Pharm. Bull., 32, 3968 (1984).
• the 4-nitro-naphthalene-1-carboxylic acid can be prepared according to the method described by GJ. LEUCK and R.P. PERKINS, J. Amer. Chem. Soc., 5J, 1831 (1929).
• the products of general formula (I) can be purified according to the usual methods such as chromatography.
• 4-Nitro-naphthalene-1-carboxylic acid is prepared according to the method of GJ. LEUCK and R.P. PERKINS, J. Amer. Chem. Soc, 51, 1831 (1929).
• the organic solution is washed twice with 200 cm3 of an aqueous solution of sodium bicarbonate at 10% (w / v) then with an aqueous solution of citric acid at 10% (w / v), with water then distilled with a saturated aqueous solution of sodium chloride.
• the organic phase is dried over magnesium sulfate, filtered and then concentrated to dryness under reduced pressure. 24.5 g of an oil are obtained which is purified by chromatography on silica eluting with a cyclohexane-ethyl acetate mixture (1-1 by volume). 19 g of the methyl ester of N - [(4-nitro-naphthyl) -l-carbonyl] -L-methionine are thus obtained in the form of an orange oil.
• EXAMPLE 3 The methyl ester of N - [(4-nitro-naphthyl) -l-thiocarbonyl] -L- methionine is prepared, with a yield of 35%, using LAWESSON reagent, according to the OCAIN method and RICH., J. Med. Chem., 1988, 31 (11), 2195 (1988), starting from the methyl ester of N - [(4-nitro-naphthyl) -l-carbonyl] -L-methionine.
• ditrifluoroacetate of the methyl diester of di- ⁇ N- [4- (2 (R) -amino-3-mercapto-propylamino) -naphthyl-1-thiocarbonyl] -L-methionine ⁇ in the form of a powder.
• the ditrifluoroacetate of the methyl diester of di- ⁇ N- [4- (2 (R) -amino-3-mercapto-propylamino) -naphthyl- 1 -thiocarbonyl] -L-methionine ⁇ has the following characteristics:
• 6-nitro-naphthalene-1-carboxylic acid and 3-nitro-naphthalene-1-carboxylic acid are prepared in a ratio of 80/20 according to the method of T. NAKAYAMA and. coll., Chem. Pharm. Bull., 32, 3968 (1984).
• the organic solution is washed twice with 200 cm3 of an aqueous solution of sodium bicarbonate at 10% (w / v) then with an aqueous solution of citric acid at 10% (w / v), with water then distilled with a saturated aqueous solution of sodium chloride.
• the organic phase is dried over magnesium sulfate, filtered and then concentrated to dryness under reduced pressure. 13.3 g of an oil are obtained which is purified by chromatography on silica eluting with a cyclohexane-ethyl acetate mixture (1-1 by volume).
• the reaction mixture is poured onto ice and then brought to a pH close to 7-8 by addition of an aqueous solution of sodium hydrogencarbonate at 5% (w / v).
• the mixture obtained is filtered on sintered glass garnished with celite.
• the organic phase is separated by decantation and the aqueous phase is extracted 3 times with 400 cm3 of ethyl acetate.
• the organic phases are combined, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure. 3 g of an oil are thus obtained which is purified by chromatography on silica eluting with a cyclohexane-ethyl acetate mixture (1-1 by volume).
• reaction mixture is stirred for 48 hours at a temperature in the region of 20 ° C. and then filtered through sintered glass lined with celite. The sintered glass is washed with acetonitrile. The filtrate is concentrated under reduced pressure. An orange oil is obtained which is purified by chromatography on silica eluting with a cyclohexane-ethyl acetate mixture (7-3 by volume). 1.67 g of a mixture of methyl esters of N- [6- (2 (R) -tert-butoxycarbonylamino-3-triphenylmethylthio-propylamino) -naphthyl-1-carbonyl] -L-methionine and N- are obtained.
• the reaction mixture is stirred for 2 hours at a temperature in the region of 20 ° C., then concentrated under reduced pressure.
• the residue is triturated 3 times with 15 cm 3 of ethyl ether and then dried under reduced pressure.
• the residue is purified by high performance liquid chromatography (phase Cl 8), eluting with xin acetonitrile-water mixture containing 0.1% trifluoroacetic acid.
• the methyl esters of N - [(6-nitro-naphthyl) -l- thiocarbonyl] -L-methionine and N - [(3-nitro-naphthyl) -l-thiocarbonyl] -L-methionine are prepared. in a ratio 70-30, with a yield of 50%, by the LAWESSON reagent, according to the method of OCAIN and RICH., J. Med.
• the reaction mixture is poured onto ice and then brought to a pH close to 7-8 by addition of an aqueous solution of sodium hydrogencarbonate at 5% (w / v).
• the mixture obtained is filtered through a sintered glass lined with celite.
• the organic phase is separated by decantation and the aqueous phase is successively extracted 3 times with 150 cm3 of ethyl acetate.
• the organic phases are rexinized, dried over magnesium sulfate, filtered and concentrated to dryness under reduced pressure.
• reaction mixture is stirred for 24 hours at a temperature in the region of 20 ° C. and then filtered through sintered glass lined with celite. The sintered glass is washed with acetonitrile. The filtrate is concentrated under reduced pressure. A meringue is obtained which is purified by chromatography on silica eluting with a dichloromethane-ethyl acetate mixture (95-5 by volume). 1.2 g of a mixture of the methyl esters of N- [6- (2 (R) -tert-butoxycarbonylamino-3-triphenylmethylthio-propylamino) -naphthyl-1-thiocarbonyl] -L-methionine are obtained.
• the reaction mixture is stirred for 1 hour at a temperature in the region of 20 ° C., then concentrated under reduced pressure.
• the residue is successively triturated with 3 times 5 cm 3 of hexane, 3 times 5 cm 3 of pentane, 3 times 5 cm 3 of ethyl ether and then dried under reduced pressure.
• the compounds are purified and separated by high performance liquid chromatography (phase C18), eluting with an acetonitrile-water mixture containing 0.1% trifluoroacetic acid.
• the famesyltransferase activity is determined by the amount of (- ⁇ H) famesyl transferred from ( ⁇ H) famesylpyrophosphate [( ⁇ H) FPP) to the p21 H-Ras protein.
• the standard reaction mixture is composed, for a final volume of 60 ⁇ l, of Tris-HCl 50 mM, MgCl2 5 mM, dithiotreitol 5 mM, octyl- ⁇ -D-glucopyranoside 0.2%, ⁇ 21 H-ras 200 picomoles, ( 3 H ) FPP (at 61,000 dpm / picomole) 4.5 picomoles.
• the reaction is initiated by the addition of approximately 5 ng of human famesyltransferase purified from cultures of THP1 cells. After incubation for 20 minutes at 37 ° C. in a microtiter plate containing 96 holes of 1 cm 3 per plate (Titer Plate®, Beckman), the reaction is stopped by adding 0.4 cm 3 of 0.1% SDS in methanol. at 0 ° C. 0.4 cm 3 of trichloroacetic acid (TCA) 30% in methanol is then added to the mixture. The plates are left for 1 hour in ice.
• TCA trichloroacetic acid
• the precipitated content is then retained on a fiberglass membrane Filtermat®, Pharmacia) with the filtration unit (Combi Cell Harvester®, Skatron) and rinsed with 6% trichloroacetic acid in distilled water.
• the membranes are dried in the microwave oven then impregnated with scintillant by melting under hot air with Meltilex® (Pharmacia) and finally counted in cpm in a ⁇ -Plate® counter (LKB). Each test is repeated 3 times.
• the activity unit is defined by 1 picomole of ( 3 H) FPP transferred to p21 H-Ras in 20 minutes.
• the inhibition percentages are obtained by comparison of the tests with and without inhibitor after deduction of the blanks, the IC50 being measured from the inhibitions obtained with 9 different concentrations using the Enzfitter® or Graf it® software.
• the activity on cells can be determined as follows:
• the cell line is the THAC line (CCL 39 cells transfected with activated Ha-Ras) according to K. Seuwen et al., EMBO J., 7 (1) 161-168 (1988).
• the cells are seeded in 6 cm diameter petri dishes containing DMEM medium, 5% fetal calf serum, 1% G418.
• the culture medium is changed (with or without serum) and the product to be studied is added in solution in dimethylformamide (DMF), in the presence or in the absence of DTT (final concentrations of 0.5% in DMF and 0.1mM in DTT).
• DMF dimethylformamide
• the lysates are clarified by centrifugation at 4,000 rpm for 10 minutes.
• the Ras protein is revealed by the Western blot technique: the membrane is incubated with a specific anti-Ras monoclonal antibody (pan-Ras Ab3, Oncogene Science) then with protein A labeled with 125 ⁇ l. After autoradiography, the bands are identified, cut and counted in a ⁇ counter. The radioactivity of the bands corresponding to famesylated Ras and to non-famesylated Ras makes it possible to determine the percentage inhibition of famesylation of the Ras protein. The results obtained are collated in Table I.
• the new products of general formula (I) can be in the form of non-toxic and pharmaceutically acceptable salts.
• These non-toxic salts include the salts with mineral acids (hydrochloric, sulfuric, hydrobromic, phosphoric, nitric) or with organic acids (acetic, propionic, succinic, maleic, hydroxymaleic, benzoic, fumaric, methane sulfonic, trifluoroacetic or oxalic ) or with mineral (soda, potash, lithine, lime) or organic (tertiary amines such as triethylamine, piperidine, benzylamine) bases depending on the nature of the Rjet R symbols of the product of general formula (I).
• the present invention also relates to pharmaceutical compositions which contain at least one product of general formula (I) in combination with one or more diluents or pharmaceutically acceptable adjuvants whether inert or physiologically active.
• compositions can be administered orally, parenterally or rectally.
• compositions for oral administration include tablets, pills, powders or granules.
• the active product according to the invention is mixed with one or more inert diluents such as sucrose, lactose or starch.
• these compositions can include substances other than diluents, for example a lubricant such as magnesium stearate.
• liquid compositions for oral administration can be used pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs containing inert diluents such as water or paraffin oil.
• These compositions can also include substances other than diluents, for example wetting, sweetening or flavoring products.
• compositions according to the invention for parenteral administration can be sterile aqueous or non-aqueous solutions, suspensions or emulsions.
• solvent or vehicle propylene glycol, a polyethylene glycol, vegetable oils, in particular olive oil or injectable organic esters, for example ethyl oleate.
• These compositions can also contain adjuvants, in particular wetting agents, emulsifiers and dispersants. Sterilization can be done in several ways, for example using a bacteriological filter, incorporating sterilizing agents into the composition 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.
• compositions for rectal administration are suppositories which may contain, in addition to the active product, excipients such as cocoa butter.
• excipients such as cocoa butter.
• the compositions according to the invention are particularly useful in human therapeutics in the treatment of cancers of various origins.
• the doses depend on the desired effect, the duration of the treatment and the factors specific to the subject to be treated.
• the doses are, in humans, between 0.1 and 20 mg / kg per day intraperitoneally.
• Example 2 200 mg of the product obtained in Example 1 are dissolved in 100 cm 3 of physiological saline. The solution obtained is aseptically distributed in 10 cm 3 ampoules. The ampoules are administered as a single injection or as an infusion.

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• 1995
  • 1995-07-12 FR FR9508423A patent/FR2736638B1/fr not_active Expired - Fee Related
• 1996
  • 1996-07-05 ZA ZA965750A patent/ZA965750B/xx unknown
  • 1996-07-10 EP EP96924961A patent/EP0842150A1/de not_active Ceased
  • 1996-07-10 CA CA002224411A patent/CA2224411A1/fr not_active Abandoned
  • 1996-07-10 WO PCT/FR1996/001071 patent/WO1997003047A1/fr not_active Application Discontinuation
  • 1996-07-10 AU AU65233/96A patent/AU6523396A/en not_active Abandoned
  • 1996-07-10 JP JP9505563A patent/JPH11508911A/ja active Pending
• 1998
  • 1998-01-09 US US09/005,154 patent/US5889053A/en not_active Expired - Fee Related

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