Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
  • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
  • C07D487/04—Ortho-condensed systems
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/14—Drugs for dermatological disorders for baldness or alopecia
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P33/00—Antiparasitic agents
• • 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

• CDK Cyclin-dependent kinase
• GSK-3 glycogen svnthase kinase-3
• the subject of the present invention is new inhibitors of cyclin-dependent kinases, and in particular of cyclin B1 / cdc2, and of glycogen synthase kinase-3 (GSK-3).
• CDKs cyclin-dependent kinases
• CDK1-X The enzyme cdc2 is also called CDK1 (Meijer et al., Ewr. /. Biochem. (1997), 243, 527-536).
• CDK3 each of the CDKs is specifically associated with one or more members of the cyclin family:
• CDK9 - cyclin T: CDK9.
• the CDK partners of cyclins F, G and I have not yet been identified.
• Other kinases close to cdc2 and other cyclins have been identified and the characterization of their functions is in progress, for example in parasites (Le Roch et al., /. Biol. Chem. (2000), 275, 8952 -8958) or in herpes viruses (Card et al., EMBO (2000), 19, 2877-2888).
• CDKs are controlled by two other families of enzymes which work in opposition (Jessus and Ozon, Prog. Cell Cycle Res. (1995), 1, 215-228).
• the first groups kinases such as Weel and Mikl which deactivate CDKs by phosphorylating certain amino acids (Den Haese et al., Mol. Biol. Cell (1995), 6, 371-385).
• the second groups together phosphatases such as the Cdc25 family which activates certain CDKs by dephosphorylating the tyrosine and threonine residues of CDKs (Gould et al., Science (1990), 250, 1573-1576).
• CDKs Many compounds blocking the kinase activity of CDKs are known (Meijer and Kim, Methods Enzymol. (1997), 283, 113-128). They are being studied in several therapeutic fields such as oncology to prevent the division of tumor cells (McDonald and el-Deiry, Int. J. Oncol. (2000), 16, 871-886), neurobiology to prevent natural apoptosis or chemo-induced normal cells (e.g. neurons) (Maas et al., J. Neurochem. (1998), 70, 1401-1410; Park et al., J. Neurosci.
• Cyclin-dependent kinase inhibitors are therefore likely to be used as drugs, in particular in the treatment of the diseases / disorders described. in Meijer et al., Pharmacol. Ther. (1999); 82, 279-284, and in particular:
• neurodegenerative diseases for example tauopathies and in particular Alzheimer's disease
• CDK inhibitors • and more generally in the treatment of all diseases / disorders corresponding to reported uses for CDK inhibitors.
• the glycogen synthase kinase-3 (GSK-3) enzyme (Parker et al., Eur. J. Biochem. (1983), 130, 227-234) is a serine / threonine kinase enzyme.
• GSK-3 glycogen synthase kinase-3
• the ⁇ isoform encodes a 51 kd polypeptide.
• the ⁇ isoform codes for a 47 kd polypeptide having an amino acid homology of 85% with GSK-3 ⁇ (Woodgett, EMBO (1990), 9, 2431-2438).
• the levels of messenger expression for the ⁇ and ⁇ isoforms of GSK-3 are predominant in the testes, thymus, prostate and ovaries but low in the lung and kidney. Analysis of protein detection in different tissues shows a lack of correlation between transcription and translation (Lau et al., J. Pept. Res. (1999), 54, 85-91).
• GSK-3 is in an activated form in cells where it inhibits Glycogen synthase by direct phosphorylation (Eldar-Finkelman et al., Proc. Natl. Acad. Sci. USA (1996), 93, 10228-10233) (3) .
• Insulin inhibits GSK-3 and activates glycogen synthase.
• the inhibition of GSK-3 can be observed with other growth factors such as Y Insulin-like Growth Factor-I (IGF-I) or VEpidermal Growth Factor (EGF).
• IGF-I Insulin-like Growth Factor-I
• EGF VEpidermal Growth Factor
• GSK-3 participates in other biological processes including cell cycle control (Diehl et al., Ge ⁇ es & Dev. (1998), 12, 3499-3511), the cellular distribution of ⁇ -catenin (Yost et al., Genes & Dev. (1996), 10, 1443-1454), cell survival and activation of Nf-kappaB in the control of apoptosis (Hoeflich et al., Nature (2000), 406, 86-90), glucose metabolism (Summers et al., /. Biochem. (1999), 274,17934-17940), phosphorylation of the tau protein (Spittaels et al., J. Biol. Chem., 27 September 2000), or the dynamics of microtubules (Krylova et al., /. Cell Biol. (2000), 151 (1), 83-94).
• cell cycle control Diehl et al., Ge ⁇ es & Dev. (1998), 12, 3499-35
• lithium a therapeutic agent used in the treatment of depression for many years, which is a direct inhibitor of GSK-3; in addition to these effects on depression, lithium can modulate the proliferation of normal or tumor cells (Cui et al., Brain Res. Dev. Brain Res. (1998), 111 (2), 177-88);
• CDK inhibitors except those derived from purines, which are reported to be potent inhibitors of GSK-3 (Meijer, Supplement to Cancer Clinical Research (November 2000), 6, Proceedings of the NCI-EORTC - ACCR Symposium, 043).
• CDKs which are not inhibitors of GSK-3, such as derivatives of purines (roscovitine, olomucine, purvalanol ...) and butyrolactone.
• the specific inhibition values of different products on the two classes of enzymes are reported in Leclerc et al., /. Biol. Chem., September 2000.
• neurodegenerative disorders such as Parkinson's disease
• tauopathies pathologies where the tau protein is hyperphosphorylated as in Alzheimer's disease or certain dementias;
• triazolopyrazine derivatives of simpler structures were already used in therapy, for example as phosphodiesterase inhibitors (US patents 3,846,423 and US 3,865,824), as corticotropin releasing factor (CRF) antagonists (patent applications).
• A represents a hydrogen atom, a halogen atom, a formyl, cyano, nitro, guanidinoaminomethylenyl, (1, 3-dihydro-2-oxoindol) -3-ylidenemethyl, alkylcarbonyl, aralkylcarbonyl or heteroaralkylcarbonyl radical, or a radical
• L represents an alkylene radical and R and R are chosen
• 1 9 independently from a hydrogen atom and an alkyl radical or R and R taken together with the nitrogen atom which carries them form a heterocycle of 5 to 7 members, the complementary members being independently chosen from the group consisting of - CH 2 -, -NR -, -S- and -O-, R representing independently each time it intervenes a hydrogen atom or an alkyl radical;
• X represents a hydrogen atom, an alkylthio, aralkylthio, alkylthioxo or aralkylthioxo radical, or also a NR 4 R 5 radical in which R 4 represents an alkyl radical, a hydroxyalkyl radical, a cycloalkyl radical optionally substituted by one or more chosen radicals among the alkyl, hydroxy and amino radicals, an aralkyl radical in which the aryl radical is optionally substituted by one or more radicals chosen from a halogen atom, the cyano radical, the nitro radical and the alkyl or alkoxy radicals, or alternatively R 4 represents a heteroaryl or heteroarylalkyl radical, the heteroaryl radical of the heteroaryl or heteroarylalkyl radicals being optionally substituted by one or more alkyl radicals and R represents a hydrogen atom, or then R 4 and R 5 taken together with the nitrogen atom which carries them form a heterocycle of 5 to 7 members,
• Y represents NH or an oxygen atom
• Z represents a bond or an alkyl or alkylthioalkyl radical
• Ar represents a carbocyclic aryl radical optionally substituted from 1 to 3 times by radicals chosen independently from a halogen atom, the cyano radical, the nitro radical, an alkyl or alkoxy radical and a NR 7 R 8 radical in which R 7 and R 8 independently represent a hydrogen atom or an alkyl radical or R 7 and R 8 taken together with the nitrogen atom which carries them form a heterocycle of 5 to 7 members, the complementary members being independently chosen from the group consisting of - CH 2 -, -NR 9 -, -S- and -O-, R 9 independently representing each time that it intervenes a hydrogen atom or an alkyl radical, or Ar represents a heterocyclic aryl radical with 5 or 6 members and whose heteroatom (s) are chosen from nitrogen, oxygen or sulfur atoms, said heteroatoms possibly being oxidized (Ar can represent, for example, the oxidopyridyl radical ) and said heterocyclic aryl radical which may be optionally substituted
• CDK cyclin dependent kinases
• the compounds of general formula (I) can be used to prepare a medicament intended to inhibit both cyclin-dependent kinases (CDK) and glycogen synthase kinase-3 (GSK-3) .
• CDK cyclin-dependent kinases
• GSK-3 glycogen synthase kinase-3
• alkyl when no further details are given, is meant a linear or branched alkyl radical containing from 1 to 6 carbon atoms.
• cycloalkyl when it is not given more precision, is meant a cycloalkyl radical having 3 to 7 carbon atoms.
• carbocyclic or heterocyclic aryl when it is not given more precision, is meant a carbocyclic or heterocyclic system comprising from one to three condensed rings of which at least one is an aromatic ring, a system being said to be heterocyclic when l 'at least one of the cycles which compose it includes one or more heteroatoms (O, N or S).
• aryl when it is not given more precision, one understands a carbocyclic aryl radical.
• heteroaryl is meant a heterocyclic aryl radical.
• alkylcarbonyl aralkylcarbonyl, heteroaralkylcarbonyl, alkylthio, aralkylthio, alkylthioxo, aralkylthioxo, hydroxyalkyl, alkylthioalkyl, aralkyl, heteroaralkyl, aminoalkyl, alkylaminoalkyl and dialkylaminoalkyl meant respectively the alkylcarbonyl, aralkylcarbonyl, heteroaralkylcarbonyl, alkylthio, aralkylthio, alkylthioxo, aralkylthioxo, hydroxyalkyl , alkylthioalkyle, aralkyle, heteroaralkyle, aminoalkyle, alkylaminoalkyle and dialkylaminoalkyle whose radicals aryl, heteroaryl and alkyl have the meanings indicated previously.
• linear or branched alkyl having from 1 to 6 carbon atoms is meant in particular the methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl, pentyl, neopentyl, isopentyl, hexyl, isohexyl radicals.
• Cycloalkyl is understood to mean in particular the cyclopropyl and cyclohexyl radicals.
• carbocyclic aryl is meant in particular the phenyl and naphthyl radicals.
• heterocyclic aryl in particular the pyrrolyl, furannyl, thienyl, pyridyl, imidazolyl, oxazolyl, thiazolyl, indolyl and quinolyl.
• halogen is meant the fluorine, chlorine, bromine or iodine atoms.
• pharmaceutically acceptable salt means in particular the addition salts of inorganic acids such as hydrochloride, hydrobromide, iodhydrate, sulfate, phosphate, diphosphate and nitrate or organic acids such as acetate, maleate, fumarate, tartrate, succinate, citrate , lactate, methanesulfonate, p-toluenesulfonate, pamoate and stearate.
• inorganic acids such as hydrochloride, hydrobromide, iodhydrate, sulfate, phosphate, diphosphate and nitrate or organic acids such as acetate, maleate, fumarate, tartrate, succinate, citrate , lactate, methanesulfonate, p-toluenesulfonate, pamoate and stearate.
• bases such as sodium or potassium hydroxide.
• the compounds according to the present invention may contain asymmetric carbon atoms. Consequently, the compounds according to the present invention have two possible enantiomeric forms, that is to say the “R” and “S” configurations.
• the present invention includes the two enantiomeric forms and all combinations of these forms, including the "RS” racemic mixtures. For the sake of simplicity, when no specific configuration is indicated in the structural formulas, it should be understood that the two enantiomeric forms and their mixtures are represented.
• the compounds of general formula (I) defined above, or their pharmaceutically acceptable salts can be used to prepare a medicament intended to treat the following diseases / disorders / natural phenomena: tumor proliferation, proliferation of normal cells, l alopecia, alopecia induced by exogenous products, radio-induced alopecia, spontaneous or induced apoptosis of normal cells (ischemia), meiosis, fertilization, oocyte maturation, viral or retroviral infections ( herpes, AIDS, cytomegalo virus), neurodegenerative diseases (for example tauopathies including Alzheimer's disease), proliferation of parasites (proliferation of protozoa, for example Trypanosomes, Toxoplasmas or Plasmodium) and myopathies.
• diseases / disorders / natural phenomena tumor proliferation, proliferation of normal cells, l alopecia, alopecia induced by exogenous products, radio-induced alopecia, spontaneous or induced apoptosis of normal cells (ischemia), meiosis, fertilization
• the compounds of general formula (I) defined above, or their pharmaceutically acceptable salts can be used to prepare a medicament intended to treat the following diseases / disorders / natural phenomena: tumor proliferation, proliferation of normal cells, in particular especially restenosis, and tauopathies including Alzheimer's disease.
• the compounds according to the invention will be such that they have at least one of the following characteristics:
• A represents a hydrogen atom, a halogen atom, a formyl, cyano, nitro, guanidinoaminomethylenyl, (1,3-dihydro-2-oxoindol) -3-ylidenemethyl, alkylcarbonyl or aralkylcarbonyl radical, or also a radical -L- NR R in which L
• R and R are independently chosen from a
• X represents a hydrogen atom, an alkylthio or alkylthioxo radical, or also a NR R 5 radical in which R 4 represents an alkyl radical, a hydroxyalkyl radical, a cycloalkyl radical optionally substituted by one or more amino radicals, an aralkyl radical the aryl radical of which is optionally substituted by one or more radicals chosen from a halogen atom and the alkyl or alkoxy radicals, or alternatively R 4 represents a heteroaryl or heteroarylalkyl radical, the heteroaryl radical of the heteroaryl or heteroarylalkyl radicals being optionally substituted by one or more alkyl radicals and R 5 represents a hydrogen atom, or then R 4 and R 5 taken together with the nitrogen atom which carries them form a heterocycle of 5 to 7 members, the complementary members being chosen independently from the group composed of -CH 2 -, -NR 6 -, -S- and -O-, R 6 independently representing each time
• the compounds according to the invention will be such that they have at least one of the following characteristics:
• A represents a halogen atom, a formyl radical, guanidinoaminomethylenyl, (1,3-dihydro-2-oxoindol) -3-ylidènemethyl or alkylcarbonyl, or a radical -L-NR ⁇ 2 in which L represents a methylene radical and R 1 and R 2 are chosen independently from a hydrogen atom and an alkyl radical or R 1 and R 2 taken together with the nitrogen atom which carries them form a heterocycle of 5 to 7 members, the complementary members being chosen independently from the group consisting of -CH 2 -, -NR 3 - and -O-, R 3 representing independently each time it intervenes a hydrogen atom or an alkyl radical; • X represents an alkylthio or alkylthioxo radical, or also a NR 4 R 5 radical in which R 4 represents an alkyl radical, a hydroxyalkyl radical, a cycloalkyl radical optionally substituted by one or more
• Ar represents a carbocyclic aryl radical optionally substituted from 1 to 3 times by radicals independently chosen from a halogen atom and a radical NR 7 R 8 in which R 7 and R 8 independently represent a hydrogen atom or an alkyl or R radical 7 and R 8 taken together with the nitrogen atom which carries them form a heterocycle of 5 to 7 members, the complementary members being chosen independently from the group composed of -CH 2 -, -NR 9 - and -O-, R 9 independently representing each time that it intervenes a hydrogen atom or an alkyl radical, or alternatively Ar represents a heterocyclic aryl radical having 5 or 6 members and the heteroatom (s) of which are chosen from nitrogen atoms and d oxygen, said heteroatoms possibly being oxidized and said heterocyclic aryl radical possibly being substituted by one or more radicals chosen independently from alkyl, amine radicals oalkyl, alkylaminoalkyl and dialkylaminoalkyl.
• the compounds according to the invention will be such that they have at least one of the following characteristics:
• A represents a halogen atom, a formyl radical, guanidinoaminomethylenyl, (1,3-dihydro-2-oxoindol) -3-ylidènemethyl or alkylcarbonyl, or also a radical -L-NR J R 2 in which L represents a methylene radical and R 1 and R 2 are chosen
• R 1 9 independently from a hydrogen atom and an alkyl radical or R and R taken together with the nitrogen atom which carries them form a heterocycle of 5 to 7 members, the complementary members being independently chosen from the group consisting of - CH 2 -, -NR 3 - and -O-, R 3 representing independently each time it intervenes a hydrogen atom or an alkyl radical; • X represents an alkylthio (and preferably methylthio) or alkylthioxo (and preferably methylthioxo) radical, or also a NR 4 R 5 radical in which R 4 represents an alkyl radical, a hydroxyalkyl radical, a cycloalkyl radical (and preferably cyclohexyl ) optionally substituted by one or more amino radicals, or alternatively R 4 represents a heteroaryl or heteroarylalkyl radical, the heteroaryl radical of the heteroaryl or heteroarylalkyl radicals being optionally substituted by one or more alkyl radicals and R
• Ar represents a carbocyclic aryl radical (said carbocyclic aryl radical preferably being a phenyl radical) optionally substituted from 1 to 3 times by radicals chosen independently from a halogen atom and a radical NR 7 R 8
• R and R independently represent a hydrogen atom or an alkyl radical or R 7 and R 8 taken together with the nitrogen atom which carries them form a heterocycle of 5 to 7 members, the complementary members being chosen independently from the group consisting of -CH 2 - and -NR 9 -, R 9 representing independently each time it involves an alkyl radical, or alternatively
• Ar represents a heterocyclic aryl radical containing 5 or 6 members and of which the heteroatom (s) are chosen from nitrogen and oxygen atoms (said heterocyclic aryl radical preferably being a pyridyl radical), said heteroatoms possibly being oxidizable and said heterocyclic aryl radical possibly being optionally substituted by one or more radicals chosen independently from the radicals alkyl, aminoalkyl, alkylaminoalkyl and dialkylaminoalkyl.
• A represents a formyl, cyano, nitro, guanidinoaminomethylenyl, (1,3-dihydro-2-oxoindol) -3-ylidenemethyl, alkylcarbonyl, aralkylcarbonyl or heteroaralkylcarbonyl radical, or a radical -L-NR ⁇ 2 in which L represents a
• R and R are independently chosen from a hydrogen atom and an alkyl radical or R 1 and R 2 taken together with the nitrogen atom which carries them form a 5- to 7-membered heterocycle, the members complementary being independently selected from the group consisting of -CH 2 -, -NR 3 -, -S- and -O-, R 3 representing independently each time it intervenes a hydrogen atom or an alkyl radical;
• X represents a hydrogen atom, an alkylthio, aralkylthio, alkylthioxo or aralkylthioxo radical, or also a NR 4 R radical in which R 4 represents an alkyl radical, a hydroxyalkyl radical, a cycloalkyl radical optionally substituted by one or more radicals chosen from alkyl, hydroxy and amino radicals, an aralkyl radical in which the aryl radical is optionally substituted by one or more radicals chosen from a halogen atom, the cyano radical, the nitro radical and the alkyl or alkoxy radicals, or alternatively R represents a heteroaryl radical or heteroarylalkyl, the heteroaryl radical of the heteroaryl or heteroarylalkyl radicals being optionally substituted by one or more alkyl radicals and R 5 represents a hydrogen atom, or then R 4 and R 5 taken together with the nitrogen atom which carries them form a 5- to 7-membered heterocycle,
• Z represents a bond or an alkyl or alkylthioalkyl radical
• Ar represents a carbocyclic aryl radical optionally substituted from 1 to 3 times by radicals chosen independently from a halogen atom, the cyano radical, the nitro radical, an alkyl or alkoxy radical and a NR 7 R 8 radical in which R 7 and R 8 independently represent a hydrogen atom or an alkyl radical or R 7 and R 8 taken together with the nitrogen atom which carries them form a heterocycle of 5 to 7 members, the complementary members being independently chosen from the group consisting of - CH 2 -, -NR 9 -, -S- and -O-, R 9 independently representing each time that it intervenes a hydrogen atom or an alkyl radical, or alternatively Ar represents a heterocyclic aryl radical having 5 or 6 links and the heteroatom (s) of which are chosen from nitrogen, oxygen or sulfur atoms, said heteroatoms possibly being oxidized (Ar may represent, for example, the oxidopyridyl radical) and said aryl radical heterocyclic which may be optionally
• Z represents an alkyl or thioalkyl radical
• X represents a radical NR 4 R 5 in which R 4 represents an aralkylthio, aralkylthioxo or hydroxyalkyl radical, one of the alkyl, alkylthio or alkylthioxo radicals containing from 2 to 5 carbon atoms, a cycloalkyl radical optionally substituted by one or radicals chosen from alkyl, hydroxy and amino radicals, an aralkyl radical in which the aryl radical is optionally substituted by one or more radicals chosen from a halogen atom and the alkyl or alkoxy radicals, or alternatively R 4 represents a heteroaryl or heteroarylalkyl radical, the heteroaryl radical of the heteroaryl or heteroarylalkyl radicals being optionally substituted by one or more alkyl radicals and R 5 represents a hydrogen atom, or else R 4 and R 5 taken together with the nitrogen atom which carries them form a heterocycle of 5 to 7 links, the complementary links being chosen independently from the
• the invention relates, as medicaments, to the compounds of Examples 3 to 33.
• compositions comprising, as active principle, at least one compound of general formula (II), one of the compounds of Examples 3 to 33 or a pharmaceutically acceptable salt of one of the latter.
• compounds of general formula (II) or their salts or one of the compounds of Examples 3 to 33 or a salt of one of these are also relates to pharmaceutical compositions.
• compositions containing a compound of the invention can be in the form of a solid, for example powders, granules, tablets, capsules, liposomes or suppositories.
• Suitable solid carriers can be, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, carboxymethyl cellulose sodium, polyvinylpyrrolidine and wax.
• compositions containing a compound of the invention can also be presented in liquid form, for example, solutions, emulsions, suspensions or syrups.
• suitable liquid carriers can be, for example, water, organic solvents such as glycerol or glycols, as well as their mixtures, in varying proportions, in water.
• the administration of a medicament according to the invention can be done by topical, oral, parenteral route, by intramuscular injection, etc.
• the administration dose envisaged for a medicament according to the invention is between 0.1 mg to 10 g depending on the type of active compound used.
• the compounds of general formula (I) can be prepared by the methods described below. Preparation of the compounds of general formula (I):
• a number of triazolopyrazines of general formula (I) can be easily prepared according to the procedures described in US Patent 4,565,815.
• the other compounds of general formula (I) according to the invention can be prepared in a few steps, scheme 1, from the compounds of general formula (III) in which A 'represents a hydrogen atom or a halogen atom and X' represents a hydrogen atom or an alkylthio radical.
• scheme 1 The preparation of the compounds of general formula (III) is described in US Patent 4,565,815 or in Kobe et al., J. Het. Chem. (1974), 11 (2), 199 and s.
• the starting compound of general formula (III) is such that X represents H or alkylthio and A represents H or a halogen atom Hal.
• the synthesis strategy is summarized in diagram 2 below.
• the compound of general formula (III) is subjected to a nucleophilic substitution reaction with the compound of general formula (IV) to give the compound of general formula (I).
• the reaction can, if necessary, be carried out in a solvent such as chloroform.
• the starting compound of general formula (III) is such that X 'represents alkylthio and preferably methylthio.
• Diagram 3 The compound of general formula (III) is first subjected to a substitution reaction with alcohol or the amine of general formula (IV) to give the compound of general formula (V).
• the compound of general formula (V) is then treated with rnet ⁇ -chloroperbenzoic acid and then with the amine of general formula R 4 NHR 5 to finally give the compound of general formula (I).
• These reactions are preferably carried out in a solvent such as chloroform.
• the compound of general formula (VI) represented in scheme 4 is used as starting material.
• This compound is a compound of general formula (I) in which A represents H and its synthesis has therefore been described previously.
• the compound of general formula (VI) is for example first treated with an excess of (chloromethylene) -dimethylammonium chloride in an aprotic polar solvent such as an acetonitrile-dimethylformamide mixture. This makes it possible to obtain compounds of general formula (I) in which A represents the formyl radical.
• These compounds can be prepared in a conventional manner from the compound of general formula (VI), for example according to the process represented in scheme 5.
• the compound of general formula (VI) can for example be treated at low temperature (for example at -78 ° C) successively with butyllithium in a polar aprotic solvent such as ethyl ether or tetrahydrofuran then the compound of general formula (VII) in which Hal represents a halogen atom, before being hydrolyzed with water slightly acidified to give the compound of general formula (I) in which A represents a radical -L-NR ⁇ 2 .
• the compound of general formula (I) in which A represents a formyl radical is converted into the compound of general formula (I) in which A represents a guanidinoaminomethylenyl radical, scheme 7, by reaction with the aminoguanidine bicarbonate in a solvent such as ethanol and in the catalytic presence of a base like piperidine.
• the compound of general formula (I) in which A represents a formyl radical is converted into the compound of general formula (I) in which A represents a (1,3-dihydro-2-oxoindol) -3-ylidenemethyl radical by the same type of reaction, oxoindole replacing bicarbonate of aminoguanidine.
• the compound of general formula (I) in which A represents a formyl radical is converted into the compound of general formula (I) in which A represents a cyano radical, scheme 8, by reaction with hydroxylamine in a mixture of sodium formate and d formic acid. The reaction is preferably carried out with heating.
• Example 12 8-bromo-2- (1R-isopropyl-2-hydroxyethylamino) - 4- (3-chloroanilino) -pyrazolo [1,5-a] -1-3-5-triazine
• the reaction medium is diluted with chloroform (10 ml) and is washed with an aqueous solution of NaHSO 3 then an aqueous solution of NaHCO 3 .
• Example 13 8-bromo-2- (2-aminocyclohexylamino) -4- (3-chloroanilino) - pyrazolo [1,5-a] -1,3,5-triazine
• Example 14 8-bromo-2- (1R-isopropyl-2-hydroxyethylamino) -4- (3-oxido-pyridylmethylamino) -pyrazolo [1,5-a] -1, 3-5-triazme
• Example 16 8-bromo-2- (4 '-hydroxy ethylpiperazinyl) -4- (3-oxido-pyridylmethylamino) -pyrazolo [1-5-a] -1, 3-5-triazme
• Example 17 8-bromo-2- (4'-hydroxyethylpiperazinyl) -4- (3-pyridylmethylamino) - pyrazolo [1,5-a] -1, 3-5-triazine
• Example 20 8-acetyl-4- (3-pyridylmethyIamino) -2-methyIthiopyrazolo [1,5-a] - 1,3,5-triazine
• A1C1 3 are successively added to a solution of 2-methylthio-4- (3-pyridylmethylamino) -pyrazolo- [1,5-a] - 1,3,5-triazine (110 mg) in 15 ml of dichloromethane then 90 ⁇ l of acetyl chloride. The mixture is brought to reflux for 4 hours. After dilution with chloroform (20 ml), the mixture is acidified with dilute HCl, then basified with an aqueous solution of NaHCO 3 and the organic phase recovered is dried over MgSO 4 . The solvents are removed by dry evaporation under vacuum.
• Example 24 8 - [(1,3-dihydro-2-oxoindol) -3-ylidenemethyl) -2-methylthio- 4- (3-pyridylmethylammo) pyrazolo [1,5-a] -1-3-5-triazine
• Example 25 8- (guanidmoammomethylene) -2-methylthio- 4- (3-pyridylmethylamino) pyrazoIo [1-5-a] -1-3.5-triazine
• This compound is prepared according to a procedure analogous to that described for intermediate 12.1. Dark yellow powder. Melting point: 70-71 ° C.
• Example 28 8 - [(1,3-dihydro-2-oxoindol) -3-ylidèneméthyI] -2-methylthio- 4- [3- (l-imidazoIyI) propylammo] pyrazolo [1,5-a] -1, 3-5-triazine
• This compound is prepared according to a procedure analogous to that described for Example 24, 8-formyl-2-methylthio-4- (3-pyridylmethylamino) pyrazolo [1,5-a] - 1,3,5-triazine being replaced by 8-formyl-2-methylthio-
• This compound is prepared by heating at reflux a mixture containing the compound of Example 22 (1 equivalent), hydroxylamine hydrochloride (2 equivalents), sodium formate (10 equivalents) and formic acid (100 equivalents) ) (cf. /. Chem. Soc. (1965), 1564). Solid pale yellow. Mass spectrometry (Electrospray): 298.2.
• This compound is prepared according to a procedure analogous to that described for Example 23, the morpholine being replaced by N-methylpiperazine. Solid brown. Mass spectrometry (Electrospray): 385.4; 386.4.
• Cupric nitrate (70 mg) is added to a suspension of 2-methylthio- 4- (3-pyridylmethylamino) -pyrazolo [1,5-a] -1,3,5-triazine (50 mg; compound of the example 31) in 6 ml of acetic anhydride.
• the mixture is stirred at ambient temperature overnight before being partitioned between chloroform and a saturated aqueous solution of NaHCO 3 .
• the organic phase is dried over MgSO 4 and the solvents are evaporated to dryness under vacuum.
• the residue is subjected to preparative chromatography on silica gel using a chloroform-methanol mixture (15: 1) as eluent. The appropriate fraction is isolated and extracted with the chloroform methanol mixture.
• the cyclin B / CDK1 complex isolated from starfish oocytes (Marthasterias glacialis) is purified by affinity chromatography and then is eluted with 0.2 M NaCl. Glycerol at a final concentration of 20%, v / v is added to the purified enzyme before storage at -80 ° C (Meijer and Kim, Methods Enzymol. (1997), 283, 113-128). The reaction is carried out in 96-well plates with a final volume of 30 ⁇ ⁇ .
• Each reaction contains 5 ⁇ l of histone Hl at 5 mg / ml in final concentration (Sigma, H5505, Saint Quentin en Yvelines, France), 16 ⁇ ⁇ of buffer composed of ⁇ -glycerophosphate 60 mM (Sigma, G6251, Saint Quentin in Yvelines, France), p-nitrophenyl phosphate 30 mM (Sigma, N6260, Saint Quentin en Yvelines, France), MOPS 25 mM (Sigma M5789, Saint Quentin en Yvelines, France), dithiothreitol 1 mM (Sigma D9779, Saint Quentin en Yvelines , France), ⁇ GTA 5 mM (Sigma ⁇ 8145, Saint Quentin en Yvelines, France), sodium orthovanadate 0, lmM (Sigma S6508, Saint Quentin en Yvelines, France), MgCl 2 15 mM (Sigma M8286, Saint
• the inhibitor in increasing concentration is added in a volume of 3 ⁇ l.
• the reaction starts with the addition of 5 ⁇ l of an ATP solution containing 4 ⁇ l of ATPy33 (370 MBq / mmol, Amersham BFIOOO, Les Ulis, France), 90 ⁇ l of cold ATP 1 mM (Sigma A7699, Saint Quentin en Yvelines, France) diluted in 906 ⁇ l of the buffer described above.
• the plates are incubated for 10 minutes at 30 ° C.
• the reaction medium is recovered on 96-well phosphocellulose P81 filtration plates (Unifilter Polyfiltronics Whatman 7700-0512, Rungis, France) and washed with 1% TCA on a collector (Filtermate Harvester, Packard, Rungis, France). After the filter has dried, Microscint ® 0 scintillant (Packard, 6016311, Rungis, France) is distributed to all wells. The radioactivity is read in a scintillation counter for Topcount ® microplates (Packard, Rungis, France). The results are expressed as the value of the concentration of inhibitor inhibiting 50% of the enzymatic reaction.
• the effect of a treatment on two human cell lines Mia-Paca2 and DU145 will be studied with the compounds of examples 1 to 33 described above.
• the DU145 (human prostate cancer cells) and Mia-PaCa2 (human pancreatic cancer cells) cell lines were acquired from the American Tissue Culture Collection (Rockville, Maryland, USA).
• the cells were treated on day 1 for 96 hours with 5 ⁇ l of each of the compounds at increasing concentrations from 0 to 25 ⁇ M in final concentration.
• the quantification of cell proliferation is evaluated by colorimetric test based on the cleavage of the tetrazolium salt WST1 by mitochondrial dehydrogenases in viable cells leading to the formation of formazan (Boehringer Mannheim, Meylan, France ). These tests are carried out in duplicate with 8 determinations per concentration tested.
• the values included in the linear part of the sigmoid were used for a linear regression analysis and used to estimate the inhibitory concentration IC 50 .
• the products are dissolved in dimethylsulfoxide (DMSO) at 10 "2 M and used in culture with 0.5% DMSO in the end. Results:
• the compounds of the present invention were further tested according to the tests relating to the antiproliferative activity described above, their activities being compared to that of Roscovitine. All the compounds tested of the present invention showed an antiproliferative activity superior to that of Roscovitine with regard to Mia-PaCa2 cells. In addition, while no antiproliferative activity with respect to DU-145 cells was observed with Roscovitine, many of the compounds tested of the present invention showed antiproliferative activity against these cells as well.

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