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WO2006005725A2 - Composes organiques - Google Patents

Composes organiques Download PDF

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Publication number
WO2006005725A2
WO2006005725A2 PCT/EP2005/053271 EP2005053271W WO2006005725A2 WO 2006005725 A2 WO2006005725 A2 WO 2006005725A2 EP 2005053271 W EP2005053271 W EP 2005053271W WO 2006005725 A2 WO2006005725 A2 WO 2006005725A2
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alkyl
hydrogen
halogen
heterocyclyl
crc
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PCT/EP2005/053271
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English (en)
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WO2006005725A3 (fr
Inventor
Peter Herold
Robert Mah
Vincenzo Tschinke
Christoph Schumacher
Aleksandar Stojanovic
Michael Quirmbach
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Speedel Experimenta Ag
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Publication of WO2006005725A2 publication Critical patent/WO2006005725A2/fr
Publication of WO2006005725A3 publication Critical patent/WO2006005725A3/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems

Definitions

  • the invention relates to novel heterocycles, to a process for the preparation of the compounds according to the invention, to pharmaceutical preparations comprising them and to their use as pharmaceutical active substances, in particular as aromatase inhibitors.
  • the present invention relates first to compounds of the general formula
  • W represents C or, if Z represents a bond and X represents C, also represents N;
  • X represents C or, if Z represents a bond, also represents N;
  • Y represents C or, if Z represents C, also represents N;
  • Z represents C or a bond
  • R represents hydrogen, d-C ⁇ -alkyl, Ci-C 8 -alkoxy-C 0 -C 4 -alkyl, halogen or trifluoromethyl;
  • R 1 represents hydrogen, Ci-C 8 -alkyl, Co-C 8 -alkylcarbonyl, carbamoyl, mono- or di(CrC 8 - alkyl)aminocarbonyl, carboxy, carboxy-Ci-C 4 -alkyl, halogen, cyano, trifluoromethyl,
  • C 8 -alkylcarbonylamino C 0 -C 8 -alkylcarbonyl-Ci-C 8 -alkylamino, carbamoyl, mono- or di(Ci-C 8 - alkyl)aminocarbonyl, carboxy-C 0 -C 4 -alkyl, Ci-C 8 -alkoxy, CrC ⁇ -alkoxycarbonyl, aryl or heterocyclyl, and, in addition, if W represents C, also represents amino, mono- and di(Ci-C 8 - alkyl)amino, Co-C ⁇ -alkylcarbonylamino, C 0 -C 8 -alkylcarbonyl-CrC 8 -alkylamino or C 1 -C 8 - alkoxy-Co-C 4 -alkyl;
  • R 2 represents heterocyclyl or aryl, which radicals are unsubstituted or substituted by 1-4
  • L represents -C(O)-, -C(R 3 )(R 4 )-O-, -C(R 3 )(R 4 )-S(O) m - or -C(R 3 )(R 4 )-NR 5 - and, in addition, if W represents C, also represents -O-, -NR 5 -, -S(O) n ,-, -O-C(R 3 )(R 4 )-, -S(O) m -C(R 3 )(R 4 )-,
  • R 3 represents hydrogen or Ci-C 8 -alkyl
  • R 4 a) represents hydrogen or C r C 8 -alkyl; or b) represents, together with R 3 , oxo;
  • R 5 represents hydrogen, Ci-C 8 -alkyl or C 0 -C 8 -alkylcarbonyl;
  • m represents a number 0, 1 or 2;
  • aryl denotes an aromatic hydrocarbon which generally comprises 5-14, preferably 6-10, carbon atoms and is, for example, phenyl, indenyl, e.g. 2- or 4-indenyl, or naphthyl, e.g. 1- or 2-naphthyl.
  • the abovementioned radicals may be unsubstituted or substituted one or more times, e.g. once or twice, it being possible for the substituent to be in any position, e.g.
  • Aryl-C 0 -C 4 -alkyl is, for example, phenyl, naphthyl or benzyl.
  • heterocyclyl denotes a saturated or unsaturated, 4-8-membered, particularly preferably 5-6-membered, monocyclic ring system, a saturated or unsaturated, 7-12- membered, particularly preferably 9-10-membered, bicyclic ring system and also a saturated or unsaturated, 7-12-membered, tricyclic ring system, in each case comprising an N, O or S atom in at least one ring, it also being possible for an additional N, O or S atom to be present in one ring.
  • the abovementioned radicals may be unsubstituted or substituted one or more times, e.g. once or twice, it also being possible for a plurality of identical or different substituents to be present.
  • Unsaturated monocyclic heterocyclyl-C 0 -C 4 -alkyl is, for example, pyrrolyl, pyridyl, thiophenyl, thiazolyl or oxazolyl.
  • Saturated monocyclic heterocyclyl-C 0 -C 4 -alkyl is, for example, pyrrolidinyl.
  • Unsaturated bicyclic heterocyclyl-C 0 -C 4 -alkyl is, for example, 4,5,6,7-tetrahydroisobenzofuranyl, 4,5,6,7-tetrahydrobenzothiazolyl, benzofuranyl, benzothiophenyl, indazolyl, indolyl, isoquinolinyl, quinolinyl, 3,4-dihydro-2H- [1 ,6]naphthyridinyl or pyrrolo[3,2-c]pyridin-1-yl.
  • Ci -C 8 -Al kyl may be straight-chain or branched and/or bridged and is, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or a pentyl, hexyl or heptyl group.
  • d-C ⁇ -Alkoxy is, for example, CrCs-alkoxy, such as methoxy, ethoxy, propyloxy, isopropyloxy, butyloxy, isobutyloxy, sec-butyloxy, tert-butyloxy or pentyloxy, but can also be a hexyloxy or heptyloxy group.
  • Ci-C 8 -Alkoxy-C 0 -C 4 -alkyl is, in addition to the meanings mentioned for Ci-C 8 -alkoxy, for example, Ci-C 5 -alkoxy-CrC 4 -alkyl, such as methoxyethyl, ethoxyethyl, propyloxymethyl, isopropyloxybutyl, butyloxymethyl, isobutyloxyethyl, sec-butyloxypropyl, tert-butyloxybutyl or pentyloxymethyl, but can also be a hexyloxymethyl or heptyloxymethyl group.
  • Ci-C 5 -alkoxy-CrC 4 -alkyl such as methoxyethyl, ethoxyethyl, propyloxymethyl, isopropyloxybutyl, butyloxymethyl, isobutyloxyethyl, sec-butyloxypropyl,
  • Ci-C 8 -Alkoxycarbonyl is preferably CrC ⁇ -alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl, propyloxycarbonyl, isopropyloxycarbonyl, butyloxycarbonyl, isobutyloxycarbonyl, sec-butyloxycarbonyl or tert-butyloxycarbonyl.
  • Co-Cs-Alkylcarbonyl is, for example, formyl, acetyl, propionyl, propylcarbonyl, isopropylcarbonyl, butylcarbonyl, isobutylcarbonyl, sec-butylcarbonyl or tert-butylcarbonyl.
  • Halogen is, for example, fluorine, chlorine, bromine or iodine.
  • Carboxy-Ci-C 4 -alkyl is, for example, carboxymethyl, 2-carboxyethyl, 2- or 3-carboxypropyl, 2-carboxy-2-methylpropyl, 2-carboxy-2-ethyl butyl or 4-carboxybutyl, in particular carboxymethyl.
  • Mono- or di(CrC 8 -alkyl)amino is, for example, Ci-C 4 -alkylamino, such as methylamino, ethylamino, propylamino or butylamino, or di(Ci-C 4 -alkyl)amino, such as dimethylamino, N-methyl-N-ethylamino, diethylamino, N-methyl-N-propylamino or N-butyl-N-methylamino.
  • Ci-C 4 -alkylamino such as methylamino, ethylamino, propylamino or butylamino
  • di(Ci-C 4 -alkyl)amino such as dimethylamino, N-methyl-N-ethylamino, diethylamino, N-methyl-N-propylamino or N-butyl-N-methylamino.
  • Mono- or di(CrC 8 -alkyl)aminocarbonyl is, for example, Ci-C 4 -alkylaminocarbonyl, such as methylaminocarbonyl, ethylaminocarbonyl, propylaminocarbonyl or butylaminocarbonyl, or di(C r C 4 -alkyl)aminocarbonyl, such as dimethylaminocarbonyl, N-methyl-N-ethylamino- carbonyl, diethylaminocarbonyl, N-methyl-N-propylaminocarbonyl or N-butyl-N-methylamino- carbonyl.
  • Ci-C 4 -alkylaminocarbonyl such as methylaminocarbonyl, ethylaminocarbonyl, propylaminocarbonyl or butylaminocarbonyl
  • di(C r C 4 -alkyl)aminocarbonyl such as dimethylamin
  • Co-C ⁇ -Alkylcarbonylamino is, for example, formylamino, acetylamino, propionylamino, propylcarbonylamino, isopropylcarbonylamino, butylcarbonylamino, isobutylcarbonylamino, sec-butylcarbonylamino or tert-butylcarbonylamino.
  • Co-C ⁇ -Alkylcarbonyl-CrCs-alkylamino is, for example, formyl-, acetyl-, propionyl-, propylcarbonyl-, isopropylcarbonyl-, butylcarbonyl-, isobutylcarbonyl-, sec-butylcarbonyl- or tert-butylcarbonyl-methylamino, formyl-, acetyl-, propionyl-, propylcarbonyl-, isopropylcarbonyl-, butylcarbonyl-, isobutylcarbonyl-, sec-butylcarbonyl- or tert-butylcarbonyl- ethylamino, formyl-, acetyl-, propionyl-, propylcarbonyl-, isopropylcarbonyl-, butylcarbonyl-, isobutylcarbonyl-, sec-butylcarbonyl-
  • Preferred compounds of the formula (I) are compounds of the general formulae
  • L preferably represents -C(O)-, -O-, -O-C(R 3 )(R 4 )-, -C(R 3 )(R 4 )-O- or -C(R 3 )(R 4 )-NR 5 -, very particularly preferably -C(O)-, -O- or -C(R 3 )(R 4 )-NR 5 -, in which R 5 is hydrogen or Ci-C 3 -alkyl.
  • R preferably represents hydrogen, CrC 8 -alkyl ( halogen or trifluoromethyl, very particularly preferably hydrogen or methyl.
  • R 1 preferably represents hydrogen, d-Cs-alkyl, halogen, heterocyclyl-Co-C 4 -alkyl or aryl-Co-C 4 -alkyl, very particularly preferably hydrogen, halogen or Ci-C 3 -alkyl.
  • R 2 preferably represents heterocyclyl or aryl, which radicals can be monosubstituted by halogen, cyano or heterocyclyl in particular.
  • R 3 and R 4 together preferably represent oxo.
  • R 5 preferably represents hydrogen or C r C 8 -alkyl, very particularly preferably hydrogen or Ci-C 3 -alkyl.
  • n preferably represents a number 0 or 1.
  • Preferred substituents for aryl or heterocyclyl are halogen, cyano, trifluoromethyl, heterocyclyl or C 0 -C 8 -alkylcarbonyl.
  • Very particularly preferred substituents for aryl or heterocyclyl are bromine, cyano, thiophenyl, thiazolyl, oxazolyl or acetyl.
  • L represents -C(O)-, -O- or -C(R 3 )(R 4 )-NR 5 -;
  • R represents hydrogen, C r C 8 -alkyl, halogen or trifluoromethyl, very particularly preferably hydrogen or methyl;
  • R 1 represents hydrogen, CrC 8 -alkyl, halogen, heterocyclyl-C 0 -C 4 -alkyl or aryl-Co-C 4 -alkyl, very particularly preferably hydrogen, halogen or Ci-C 3 -alkyl;
  • R 2 represents heterocyclyl or aryl, which radicals can be monosubstituted by halogen, cyano or heterocyclyl in particular;
  • R 3 represents hydrogen or Ci-C 8 -alkyl
  • R 4 a represents hydrogen or d-C 8 -alkyl; or, very particularly preferably, b) represents, together with R 3 , oxo; and R 5 represents hydrogen or Ci-C 3 -alkyl.
  • the compounds of the formula (I) having at least one asymmetric carbon atom can exist in the form of optically pure enantiomers or mixtures of enantiomers or as racemates.
  • Compounds with a second asymmetric carbon atom can exist in the form of optically pure diastereomers, mixtures of diastereomers, diastereomeric racemates or mixtures of diastereomeric racemates or as meso compounds.
  • the invention includes all these forms.
  • Mixtures of enantiomers, racemates, mixtures of diastereomeric isomers, diastereomeric racemates or mixtures of diastereomeric racemates can be separated by conventional methods, e.g. by racemate resolution, column chromatography, thin layer chromatography, HPLC and the like.
  • salts with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, nitric acid, sulphuric acid, phosphoric acid, citric acid, formic acid, maleic acid, acetic acid, succinic acid, tartaric acid, methanesulphonic acid, p-toluenesulphonic acid and the like.
  • Salts of compounds having salt-forming groups are in particular acid addition salts, salts with bases or, in the presence of a plurality of salt-forming groups, optionally also mixed salts or inner salts.
  • the compounds of the formula (I) can be prepared in a manner analogous to preparation methods disclosed in the literature. Details of the specific preparation variants can be taken from the examples.
  • the compounds of the formula (I) can also be prepared in optically pure form.
  • the separation into antipodes can be carried out by methods known per se, either, preferably, at an early stage in the synthesis by salt formation with an optically active acid, such as, for example, (+)- or (-)-mandelic acid, and separation of the diastereomeric salts by fraction crystallization or, preferably, at a rather late stage by derivatization with a chiral auxiliary component, such as, for example, (+)- or (-)-camphanoyl chloride, separation of the diastereomeric products by chromatography and/or crystallization, and subsequent cleavage of the bond to the chiral auxiliary.
  • the pure diastereomeric salts and derivatives can be analysed using current spectroscopic methods in order to determine the absolute configuration of the compound present, single-crystal X-ray spectroscopy representing a particularly suitable method.
  • Salts are primarily the pharmaceutically useful or nontoxic salts of compounds of the formula (I).
  • Such salts are, for example, formed from compounds of the formula (I) having an acid group, e.g. a carboxy or sulpho group, and are, for example, their salts with suitable bases, such as non-toxic metal salts derived from metals of Groups Ia, Ib, Na and Nb of the Periodic Table of the Elements, e.g.
  • alkali metal salts in particular lithium, sodium or potassium salts, alkaline earth metal salts, for example magnesium or calcium salts, furthermore zinc salts or ammonium salts, and also those salts which are formed with organic amines, such as optionally hydroxy-substituted mono-, di- or trialkylamines, in particular mono-, di- or tri(lower alkyl)amines, or with quaternary ammonium bases, e.g.
  • methyl-, ethyl-, diethyl- or triethylamine mono-, bis- or tris[2-hydroxy(lower alkyl)]amines, such as ethanol-, diethanol- or triethanolamine, tris(hydroxymethyl)methylamine or 2-hydroxy-tert-butylamine, N,N-di(lower alkyl)-N-[hydroxy(lower alkyl)]amine, such as N,N-dimethyl-N-(2-hydroxy- ethyl)amine, or N-methyl-D-glucamine, or quaternary ammonium hydroxides, such as tetrabutylammonium hydroxide.
  • the compounds of the formula (I) having a basic group e.g.
  • an amino group can form acid addition salts, e.g. with suitable inorganic acids, e.g. hydrohalic acid, such as hydrochloric acid or hydrobromic acid, sulphuric acid, with replacement of one or both protons, phosphoric acid, with replacement of one or more protons, e.g. orthophosphoric acid or metaphosphoric acid, or pyrophosphoric acid, with replacement of one or more protons, or with organic carboxylic, sulphonic or phosphonic acids or N-substituted sulphamic acids, e.g.
  • suitable inorganic acids e.g. hydrohalic acid, such as hydrochloric acid or hydrobromic acid, sulphuric acid, with replacement of one or both protons, phosphoric acid, with replacement of one or more protons, e.g. orthophosphoric acid or metaphosphoric acid, or pyrophosphoric acid, with replacement of one or more protons, or with organic carboxylic, sulphonic or phosphonic acids or N-
  • ⁇ - amino acids as well as methanesulphonic acid, ethanesulphonic acid, 2- hydroxyethanesulphonic acid, ethane-1 ,2-disulphonic acid, benzenesulphonic acid, 4- toluenesulphonic acid, naphthalene-2-sulphonic acid, 2- or 3-phosphoglycerate, glucose 6- phosphate or N-cyclohexylsulphamic acid (with formation of cyclamates), or with other acidic organic compounds, such as ascorbic acid.
  • Compounds of the formula (I) having acidic and basic groups can also form inner salts.
  • the compounds of the formula (I) and (la-d) also include those compounds in which one or more atoms are replaced by their stable nonradioactive isotopes; for example a hydrogen atom by deuterium.
  • Prodrug derivatives of the compounds described above are derivatives thereof which, on in vivo application, release the original compound by a chemical or physiological process.
  • a prodrug can, for example, be converted to the original compound when a physiological pH is reached or by enzymatic conversion.
  • Prodrug derivatives can, for example, represent esters of freely available carboxylic acids or S- and O-acyl derivatives of thiols, alcohols or phenols, the acyl group being as defined above.
  • ester derivatives which are converted to the original carboxylic acid by solvolysis in a physiological medium, such as, for example, lower alkyl esters, cycloalkyl esters, lower alkenyl esters, benzyl esters or mono- or disubstituted lower alkyl esters, such as lower ⁇ -(amino, mono- or dialkylamino, carboxy or lower alkoxycarbonyl)-alkyl esters or such as lower ⁇ -(alkanoyloxy, alkoxycarbonyl or dialkylaminocarbonyl)-alkyl esters; pivaloyloxymethyl esters and similar esters are conventionally used as such.
  • a physiological medium such as, for example, lower alkyl esters, cycloalkyl esters, lower alkenyl esters, benzyl esters or mono- or disubstituted lower alkyl esters, such as lower ⁇ -(amino, mono- or dialkylamino, carboxy
  • a particular compound in this invention also includes its prodrug derivative and salt form, provided that this is possible and appropriate.
  • the naturally occurring oestrogens 17 ⁇ -oestradiol (E2), oestrone (E1) and oestriol (E3) are C18 steroids derived from cholesterol. After cholesterol links up to lipoprotein receptors of steroidogenic cells, it is stored intracellularly and is brought in to sites of steroid synthesis. The aromatization of the A ring in the steroid backbone forms the final synthetic step in the formation of oestrogen.
  • oestradiol is mainly formed in the theca and granulosa cells of the ovaries and in the placenta.
  • LH luteinizing hormone
  • FSH follicle-stimulating hormone
  • oestradiol In the blood circulation, oestradiol binds reversibly to sex hormone binding giobulin, a ⁇ -giobulin, and with relatively weak affinity to albumin; 2-3% of oestradiol is nonbonded in the plasma. Oestrogens are metabolized as sulphates or giucoronides and the conjugates are removed in the gail bladder or urine. The hydrolysis of these conjugates by the intestinai flora and the resulting reabsorption can bring about an enterohepatic circulation of the oestrogens.
  • Oestrogens stimulate the growth, the blood supply and the enriching in water of the sexual organs. They can accordingly also cause breast cancer and endometrial hyperplasia. Oestrogens increase the expression of lipoprotein receptors in the liver, which results in a reduction in the level of low density lipoprotein cholesterol (LDL-C). However, they also increase the ability of the blood to clot by stimulating the production of clotting factors in the liver. In bone, the osteoclasts and osteoblasts are direct targets of oestrogens, where they exert an antiresorptive effect on the bone mass.
  • LDL-C low density lipoprotein cholesterol
  • the oestrogens stimulate the growth and the differentiation of the ductal epithelium, they induce the mitotic activity of the columnar cells of the duct and they stimulate the growth of the connective tissue. Oestrogens stimulate above all the growth of breast cancer cells. In postmenopausal women with breast cancer, the local oestradiol concentration in the tumour is increased because of increased in situ aromatase enzyme activity, in spite of low oestradiol plasma levels.
  • the chemical compounds described in the present invention exhibit useful pharmacological properties by inhibiting the aromatase enzyme (CYP19) in mammals, in particular in man. In this way, the metabolic conversion of the androgens to oestrogens is inhibited.
  • the compounds can accordingly be used for the treatment of oestrogen-dependent diseases, such as, for example, oestrogen-dependent breast cancer, above all in postmenopausal women.
  • the compounds can also, because of the inhibition of steroid aromatization, be used for the treatment of gynecomastia, the development of breast tissue in men.
  • the aromatase-inhibiting effect of these described compounds can be shown by means of in vitro assays in cell-free or cellular test systems.
  • the inhibition of the aromatase activity in vitro by the compounds described in the present invention can be shown using commercially available Cyp19 assay kits.
  • the Cyp19/Methoxy-4-trifluoromethyl-coumarin (MFC) High Throughput Inhibition Screening Kit (Becton Dickinson Biosciences, San Jose, CA, USA), for example, allows the testing in the 96-well plate format of possible chemical compounds which inhibit the Cyp19 catalytic activity.
  • the kit comprises recombinantly expressed Cyp19 enzyme in the form of supersomes, a fluorescent P450 substrate, an NADPH regenerating system, a reaction buffer and a stop reagent.
  • the MFC fluorogenic substrate is quickly converted by the Cyp19 supersome activity to the highly fluorescent product 7-hydroxy-4- trifluoromethyl-coumarin (7-HFC).
  • This assay is carried out in the presence of a wide range of concentrations of test compound, including the concentrations from 0.2 nanomolar up to 20 millimolar, according to the instructions of the manufacturer of the kit.
  • the IC 50 values for active test compounds are determined by simple linear regression analysis in order to produce inhibition curves free from data weighting.
  • the inhibition curve was calculated by fitting a 4-parameter logistic function with the raw data points using the least squares method.
  • the equation of the 4-parameter logistic function is calculated according to:
  • the compounds of the present invention show, in the in vitro test systems, inhibitory effects at minimum concentrations of approximately 10 ⁇ 3 to approximately 10 "10 mol/l.
  • the Cyp19-inhibiting properties of chemical compounds of the present invention can also be shown in a cellular test system.
  • the NCI-H295R cell line was originally derived from an adrenocortical carcinoma and has been characterized in the literature by the stimulable secretion of steroid hormones and the presence of the enzymes essential for steroid genesis.
  • the NCI-H295R cells have Cyp11 A (cholesterol side-chain cleavage), Cyp11B1 (steroid 11 ⁇ -hydroxylases), Cyp11B2 (aldosterone synthases), Cyp17 (steroid 17 ⁇ -hydroxylases and/or 17,20-lyases), Cyp19 (aromatases), Cyp21B2 (steroid 21 -hydroxylases) and 3 ⁇ -HSD (hydroxysteroid dehydrogenases).
  • the cells show the physiological property of zonal Iy undifferentiated human foetal adrenocortical cells which, however, have the ability to produce those steroid hormones which are formed in the three phenotypically distinct zones in the adult adrenalcortex.
  • the NCI-H295R cells (American Type Culture Collection, ATCC 1 Rockville, MD, USA) are cultured in Dulbecco's Modified Eagle's/Ham's F-12 Medium (DME/F12) in 75 cm 2 cell culture flasks at 37°C and in a 95% air/5% carbon dioxide atmosphere, the DME/F12 having been supplemented with Ultroser SF serum (Soprachem, Cergy-Saint-Christophe, France), insulin, transferrin, selenite (I-T-S, Becton Dickinson Biosiences, Franklin Lakes, NJ, USA) and antibiotics. The cells are subsequently transferred to a 24-well incubation flask for colony formation.
  • DME/F12 Dulbecco's Modified Eagle's/Ham's F-12 Medium
  • DME/F12 medium which now is supplemented with 0.1% bovine serum albumin instead of Ultroser SF serum.
  • the experiment is initiated by culturing the cells for 72 hours in DME/F12 medium which is supplemented with 0.1% bovine serum albumin and test compound, in the presence or absence of cell stimulants.
  • the test substance is added in a concentration range of 0.2 nanomolar up to 20 millimolar.
  • Angiotensin Il (10 or 100 nanomolar), potassium ions (16 millimolar), forskolin (10 micromolar) or a combination of two stimulants can be used as cell stimulants.
  • the cellular secretion of oestrone, oestradiol, dihydroepiandrostendione, aldosterone, corticosterone and/or Cortisol in the culture medium can be detected and quantified in radioimmunoassays using commercially available specific monoclonal antibodies according to the manufacturer's instructions. Inhibition of the release of certain steroids can be used as a measure of the respective enzyme inhibition by the test compounds added. The dose-dependent inhibition of the enzyme activity by a compound is plotted by the inhibition curve, which is characterized by an IC 50 value.
  • the IC 50 values for active test compounds are determined by simple linear regression analysis in order to produce inhibition curves free from data weighting.
  • the inhibition curve was calculated by fitting a 4-parameter logistic function with the raw data points using the least squares method.
  • the equation of the 4-parameter logistic function is calculated according to:
  • the compounds of the present invention show, in the in vitro test systems, inhibitory effects at minimum concentrations of approximately 10 "3 to approximately 10 "10 mol/l.
  • oestrogen-reducing effect of the compounds described above can be tested in vivo with suitable mammalian models, such as, for example, guinea pigs, mice, rats, cats, dogs or apes.
  • the inhibition of the aromatase activity by chemical compounds can be monitored by measuring the plasma steroid level, as described in the following protocol: female rats in the sexual cycle are treated with 5 subcutaneous injections, applied on alternating days, of 100 IU of pregnant mare serum gonadotropin (PMSG, Sigma) in 0.1 ml of sterile saline solution. 24 hours after the final injection, the test compound is administered to the animals per os at doses of 0.01 up to 10 mg/kg. Twenty-four hours after treatment, the animals are subjected to a terminal bleed. The heparinized plasma is stored at -20 0 C until analysed.
  • PMSG pregnant mare serum gonadotropin
  • the plasma levels of the steroids 17 ⁇ -oestradiol, oestrone, oestriol, progesterone, testosterone, aldosterone and corticosterone are determined using commercially available radioimmuno ⁇ assays in accordance with the instructions of the manufacturer.
  • a purification and concentration step is necessary in order to determine plasma testosterone in female rats.
  • the plasma samples are treated with 4 units by volume of diethyl ether, mixed for 15 minutes by gentle rocking and centrifuged at 2000 rpm for 5 minutes.
  • the aqueous phase is frozen with dry ice, the organic phase is separated and the solvent is evaporated using a stream of nitrogen.
  • the dry extract is again taken up in the assay buffer.
  • the inhibition of the aromatase activity by chemical compounds can also be determined by measuring the oestrogen content of the ovaries according to the following protocol: female rats, aged 21 days, are treated with a subcutaneous injection of 10 IU of pregnant mare serum gonadotropin (PMSG, Sigma). Two days later, 30 IU of human chorionic gonadotropin (hCG, Sigma) are injected subcutaneously into the same rats. On the following day, either 0.2 ml of polypropylene glycol or various doses of the test substance are injected subcutaneously into the rats. One hour later, 2.25 mg of 4-androstene-3,17-dione in 0.1 ml of oil are injected subcutaneously into the rats.
  • PMSG pregnant mare serum gonadotropin
  • hCG human chorionic gonadotropin
  • the rats are killed and their ovaries are removed.
  • the ovaries are freed from adhering tissue and stored as pairs at -50 0 C.
  • the organs are treated with 1.5 ml of a 0.05M aqueous phosphate buffer solution pH 7.4 and with 0.2 ml of a 0.1 M aqueous sodium hydroxide solution, and homogenized.
  • the homogenate is extracted with 15 ml of diethyl ether before aliquots of 5 ml are subjected to a radioimmunoassay.
  • the antiserum used for this radioimmunoassay exhibits a 100% crossreactivity for oestradiol, oestrone and oestriol.
  • the result of the analysis is expressed as ng of oestrogen content per pair of ovaries.
  • the antitumour effects of the compounds, above all for oestrogen-dependent tumours can be determined in vivo, for example in female Sprague-Dawley rats in which a breast tumour has been induced using dimethylbenzanthracene (DMBA) (see Proc. Soc. Exp. Biol. Med., 160, 296-301, 1979).
  • DMBA dimethylbenzanthracene
  • Compounds of this invention bring about, after daily application per os of doses from 1 up to 20 mg/kg or even less, a regression of existing tumours and a suppression of the emergence of new tumours.
  • the compounds of the present invention can be administered orally or enterally, for example intravenously, intraperitoneally, intramuscularly, rectally, subcutaneously or also by direct injection of the active substance locally into tissues or tumours.
  • patient describes warm-blooded animals and mammals, such as, for example, man, primates, cattle, dogs, cats, horses, sheep, mice, rats and pigs.
  • the compounds can be administered as a pharmaceutical preparation or can be incorporated in an administration device which guarantees continuous release of the compound.
  • the amount of substance to be administered can vary over a wide range and can represent any effective dose.
  • the dose of the effective substance can be between approximately 0.005 and 50 milligrams per kilogram of body weight daily; however, it is preferably between approximately 0.05 and 5 milligrams per kilogram of body weight daily.
  • the compounds can be formulated in solid or liquid pharmaceutical forms, such as, for example, as capsules, pills, tablets, sugar-coated tablets, granules, powders, solutions, suspensions or emulsions.
  • the dose of a solid pharmaceutical form can be a conventional hard gelatin capsule which can be filled with active substances and adjuvants, such as lubricants and fillers, such as, for example, lactose, sucrose and maize starch.
  • a tableting of the active substance of the present invention can represent an additional administration form.
  • the tableting can be carried out with conventional tableting adjuvants, such as, for example, lactose, sucrose or maize starch, combined with binders from gum acacia, maize starch or gelatin, disintegrating agents, such as potato starch or crosslinked polyvinylpyrrolidone (PVPP) 1 and lubricants, such as stearic acid or magnesium stearate.
  • tableting adjuvants such as, for example, lactose, sucrose or maize starch, combined with binders from gum acacia, maize starch or gelatin
  • disintegrating agents such as potato starch or crosslinked polyvinylpyrrolidone (PVPP) 1
  • lubricants such as stearic acid or magnesium stearate.
  • excipients suitable for soft gelatin capsules are vegetable oils, waxes, fats, semi-solid and liquid polyols, and the like.
  • excipients suitable for producing solutions and syrups are water, polyols, sucrose, invert sugar, glucose, and the like.
  • the compounds can be formulated in solid or liquid pharmaceutical forms, such as, for example, suppositories.
  • excipients suitable for suppositories are natural or hardened oils, waxes, fats, semi-solid or liquid polyols, and the like.
  • the compounds can be formulated as injectable doses of the active substance in a liquid or suspension.
  • the preparations generally comprise a physiologically compatible sterile solvent which can comprise a water-in-oil emulsion, with or without surfactant, and other pharmaceutically acceptable adjuvants.
  • Oils which can be used for such preparations are liquid paraffins and triglycerides of vegetable, animal or synthetic origin, such as, for example, peanut oil, soybean oil and mineral oil.
  • Injectable solutions generally comprise liquid carriers, such as, preferably, water, saline, dextrose or related sugar solutions, ethanol and glycols, such as propylene glycol or polyethylene glycol.
  • the substances can be administered as transdermal patch system, as depot injection or implant, if the formulation makes possible continuous delivery of the active substance.
  • the active substance can be compressed as granules or to narrow cylinders and can be administered subcutaneously or intramuscularly as depot injection or implant.
  • the pharmaceutical preparations can in addition also comprise preservatives, solubility promoters, viscosity-increasing substances, stabilizing agents, wetting agents, emulsifiers, sweeteners, colorants, aromatizing agents, salts for changing the osmotic pressure, buffers, coating agents or antioxidants. They can also comprise yet other therapeutically valuable substances.
  • the present invention makes possible the use of the compounds of the formula (I) and (la-d) and their pharmaceutically useable salts for the prevention, for slowing down the progression or for the treatment of symtoms or findings which respond to aromatase inhibition, especially proliferative diseases, such as breast cancer or similar types of hormone-sensitive soft tissue cancers, above all oestrogen-dependent findings, such as gynecomastia, tumours of the breast or of the endometrium, endometriosis and premature labour.
  • the compounds can also be used for the treatment or prevention of locally advanced or metastatic breast cancer in postmenopausal women who have been tested hormone receptor-positive or hormone receptor-unknown.
  • the compounds of the formula (I) and (la-d) and their pharmaceutically useable salts can be used in combination with one or more active substances with an antineoplastic effect, thus, for example, with an active substance exhibiting an antioestrogen activity, such as, for example, described for exemestane, toremifen, fulvestrant, and tamoxifen; with an active substance exhibiting an activity in inhibiting bone resorption, such as, for example, described for pamidronates and zoledronic acid; with an active substance exhibiting an alkylating activity, such as, for example, described for busulfan, temozolomide, melphalan, chlorambucile and mechlorethamine; with an active substance exhibiting an activity in the intercalation of nucleotide bases, such as, for example, described for adriamycin, daunorubicin, dactinomcyin, doxorubicin, epirubicin and idarubicin; with an active
  • a daily dose can vary within a wide range and must naturally from case to case be adjusted to the individual conditions.
  • a daily dose of 0.3 mg up to 3 g, however preferably a daily dose of 1 mg up to 1 g, is used, which, for example at a dose of 10 mg for an adult weighing 70 kg, can be administered in 1 to 3 individual daily doses proportioned identically or differently.
  • the starting materials are prepared as follows:
  • the starting material is prepared as follows:
  • the starting material is prepared as follows:

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)

Abstract

L'invention concerne de nouveaux composés hétérocycliques de formule générale (I), dans laquelle L, R, R<sup
PCT/EP2005/053271 2004-07-09 2005-07-08 Composes organiques WO2006005725A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CH1154/04 2004-07-09
CH11542004 2004-07-09

Publications (2)

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WO2006005725A2 true WO2006005725A2 (fr) 2006-01-19
WO2006005725A3 WO2006005725A3 (fr) 2006-06-01

Family

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AR (1) AR049667A1 (fr)
TW (1) TW200617010A (fr)
WO (1) WO2006005725A2 (fr)

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB8820730D0 (en) * 1988-09-02 1988-10-05 Erba Carlo Spa Substituted 5 6 7 8-tetrahydroimidazo/1.5-a/pyridines & process for their preparation
DK0625155T3 (da) * 1992-01-27 2001-11-12 Janssen Pharmaceutica Nv Pyrroloimidazolyl- og imidazopyridinyl-substituerede 1H-benzimidazolderivater som aromataseinhibitorer
CA2568159A1 (fr) * 2004-05-28 2005-12-15 Speedel Experimenta Ag Derives tetrahydro-imidazo[1,5-a] pyridine tenant lieu d'inhibiteurs de l'aldosterone syntase

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AR049667A1 (es) 2006-08-23
TW200617010A (en) 2006-06-01
WO2006005725A3 (fr) 2006-06-01

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