Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J1/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, not substituted in position 17 beta by a carbon atom, e.g. estrane, androstane
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J5/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane and substituted in position 21 by only one singly bound oxygen atom, i.e. only one oxygen bound to position 21 by a single bond
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
  • A61P15/08—Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
  • A61P15/16—Masculine contraceptives
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
  • A61P15/18—Feminine contraceptives
• • 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
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P5/00—Drugs for disorders of the endocrine system
  • A61P5/24—Drugs for disorders of the endocrine system of the sex hormones
  • A61P5/30—Oestrogens
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J7/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of two carbon atoms
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J75/00—Processes for the preparation of steroids in general

Definitions

• the present invention relates to 8 ⁇ -vinyl-11 ⁇ - ( ⁇ -substituted) alkyl-estra-1,3,5 (10) -trienes having ER ⁇ -antagonistic activity, processes for their preparation, their intermediates, pharmaceutical preparations containing the compounds according to the invention , as well as their use for the production of medicaments.
• the compounds according to the invention are those steroidal tissue-selective estrogens which have a higher affinity for estrogen receptor preparations of rat prostate than rat estrogen receptor preparations in vitro, and have a contraceptive action in vivo by virtue of their preferential effect on the ovary, and can develop through a characterized by an improved physicochemical profile.
• Contraceptive methods with chemical compounds are widely used in women who do not want to get pregnant.
• the following chemical methods of female contraception are currently available:
• Oral contraceptives which consist of various combinations of an estrogen with a progestin, are the most commonly used contraceptives in women. They work according to the endocrine principle. Although such contraceptives are very effective, undesirable side effects such as: irregular bleeding, nausea, vomiting, depression, weight gain or headache may occur. Occasionally, more severe conditions are observed, such as thromboembolism, stroke, liver adenomas, gall bladder disease or high blood pressure. These unwanted side effects of the oral contraceptives used today make clear the medical need for a new contraceptive method without side effects.
• An ideal contraceptive method is a method that attaches directly to the ovarian follicle without affecting the endocrine hypothalamic-pituitary-ovarian axis.
• Ovulation which is ovulated and fertilized, but does not lead to a preimplantation development, or c) folliculogenesis is limited and no ovulation occurs.
• Follicular growth is the development of an ovarian follicle from the primordial stage to the large ankyardous, proliferating follicle. Only an optimally constructed antral follicle has the potential to ovulate a mature egg.
• PCOS Policystic Ovary Syndrome
• Patients with impaired folliculogenesis associated with hormonal and ovulatory disorders as well as insufficiently mature oocytes (Franks et al., Mol. Cell Endocrinol 2000, 163, 49-52).
• folliculogenesis i.
• estrogen receptor ⁇ (ER ⁇ ) has been discovered as the second subtype of the estrogen receptor (Kuiper et al., Proc Natl Acad Sci 1996, 93, 5925-5930, Mosselman, Dijkema, FEBS Letters 1996, 392, 49-53 Tremblay et al., Molecular Endocrinology 1997, 11, 353-365).
• the expression pattern of ER ⁇ differs from that of ER ⁇ (Kuiper et al., Endocrinology 1996, 138, 863-870).
• ER ⁇ is only expressed in the outer follicle cells (the the the Aca cells), there is a strong expression of ER ⁇ in the oestradiol-producing granulosa cells.
• ER ⁇ and ER ⁇ Due to the different cell distribution of ER ⁇ and ER ⁇ in the ovarian follicle, it is to be expected that the interaction of a ligand with ER ⁇ or ER ⁇ will lead to different cellular responses. That ER ⁇ and ER ⁇ are functionally different has recently been confirmed by the successful production of ER ⁇ and ER ⁇ knockout mice (Couse et al., Endocrine Reviews 1999, 20, 358-417). Consequently, ER ⁇ is significantly involved in the function of the uterus, the mammary gland, the control of the sexual endocrine axis, whereas ER ⁇ is predominantly involved in the processes of ovarian physiology, in particular folliculogenesis, and ovulation.
• ER ⁇ is functional in the male animal is also evident from studies on ER ⁇ (ERKO) and ER ⁇ (BERKO) knockout mice: male ERKO mice (RA Hess et al., Nature 1997, 390, 509-512 ) show marked fertility disorders. This demonstrates the important role of estrogens in maintaining fertility testis function. ER ⁇ and ER ⁇ have significantly different amino acid sequences in their ligand binding and transactivation domain. This suggests that (1) ER subtypes with different affinity bind their ligands, and
• (2) ligands have different agonistic and / or antagonistic potential over the two receptor subtypes.
• Patent Applications WO 00/47603, WO 00/63228, WO 01/32680, WO 01/77138, US 60 / 207,370 and publications show that steroidal and nonsteroidal ligands with high affinity to ER ⁇ and ER ⁇ were found. Some compounds were considerably stronger agonists / antagonists on ER ⁇ , whereas other compounds were stronger agonists / antagonists on ER ⁇ .
• WO 00/31112 new steroidal compounds are described based on the base of the 8-position unsubstituted estradiol, which carry in 11 ß position a hydrocarbon radical containing a single linear chain with a length of 5 to 9 carbon atoms. These compounds have an ER ⁇ - agonistic / ER ⁇ antagonistic profile of action. Because of this mixed estrogen receptor profile, these compounds are useful as improved estrogens for the treatment of estrogen-related disorders and for contraception with a progestin.
• WO 01/77138 discloses 11 ⁇ -n-pentyl and 11 ⁇ -n-hexyl-8 ⁇ -substituted estra-1,3,5 (10) -trienes having ER ⁇ -antagonistic activity.
• 11 ⁇ - / --alkyl substitution leads to a further reduction of the polarity and thus also to the poorer water solubility of such compounds.
• the object of the present invention is to provide compounds with improved physicochemical properties which have a dissociation in vitro of binding to estrogen receptor preparations of rat prostate Ge025 and rat uterus and in vivo by their preferential effect on the ovary have a contraceptive effect, without other estrogen-sensitive organs such eg affecting the uterus or the liver. Furthermore, these compounds should be used for contraception in men and for the treatment of benign or malignant proliferative diseases of the ovary.
• the present invention relates to compounds of general formula I.
• R 3 is a group R 1 -O-, R u SO 2 -O-, -OC (O) R 321 1 .; n 3, 4, 5: X is a group of the formula II
• Z and W are independently R 19 , or
• Y is -OR 19 , -CN, -SCN, a halogen atom, R 20 , R 20 SO 2 -O-; or YR 19 or R 20 when Z and W together represent an oxygen atom;
• R 23 and R 24 independently represent a hydrogen atom or a halogen
• R 17 is hydrogen, -OR 19 or halogen
• alkynyl or an unsubstituted or substituted aryl, heteroaryl, heterocyclyl, aryl-C 1 -C -alkylene, heteroaryl-C 1 -C 4 -alkylene group; 20 is an R 21 R 22 N group, a group -C (NOR 19 ) H, or a group of the general formula III
• M is 0, 1, 2, 3, 4, 5, 6, 7 or 8;
• o is 0, 1, 2, 3, 4, 5, 6, 7 or 8, the sum of which is m + o 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12;
• R 21 and R 22 are independently R 19 ; R 25 R 19 , R 20 SO 2 - or an acyl group -C (O) R 21
• the present invention also encompasses the pharmaceutically acceptable salts of the compounds of general formula I according to the invention.
• the unbranched it may be, for example, a methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, ⁇ -heptyl, n-octyl; and the branched C 3 -C 8 alkyl groups a / 'so-propyl, / so-butyl, sec-butyl, te / f-butyl, / so-pentyl, neo-pentyl, 2-methylpentyl , 2,2-dimethylbutyl, 2,3-dimethylbutyl, 2-methylhexyl, 2,2-dimethylpentyl, 2,2,3-trimethylbutyl, 2,3,3-trimethylbutyl group.
• the optionally substituted with a phenyl radical C 3 -C 6 -cycloalkyl groups may be consistently a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or phenylcyclopropyl, phenylcyclobutyl, phenylcyclopentyl, Phenylcyclohexyl- group act.
• the (C 3 -C 6 -cycloalkyl) -CC-C 4 -alkylene groups may be, for example, a cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, cyclopropylethyl, cyclobutylethyl, cyclopentylethyl, cyclohexylethyl, cyclopropylpropyl, Cyclobutylpropyl, cyclopentylpropyl, cyclohexylpropyl, cyclopropylbutyl, cyclobutylbutyl, cyclopentylbutyl and cyclohexylbutyl groups, respectively.
• the branched or unbranched C 2 -C 5 -alkenyl groups may be, for example, a vinyl, trifluorovinyl, allyl, homoallyl, ( ⁇ ) -but-2-enyl, (Z) -but-2-enyl , (E) -but-1-enyl, (Z) -but-1-enyl, pent-4-enyl, (E) -pent-3-enyl, (Z) -pent-S- enyl, (E) -pent-2-enyl, (Z) -pent-2-enyl, ())) -pent-1-enyl, (Z) -penem-enyl, 2-methylvinyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, (E) -2-methylbut-2-enyl, (Z) -2-methylbut-2-enyl, 3-methylbut-2 act enyl group.
• the C 2 -C 5 -alkynyl groups may be, for example, an ethynyl, prop-1-inyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-one. inyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, 1-methylprop-2-ynyl, 1-methylbut-3-ynyl, 1 -Ethylprop-2-inyl group act.
• R 9 O groups for example, methoxy, ethoxy, n-propoxy, / 'so-propoxy, n-butoxy, sec-butoxy, / so-butoxy, tert- Butoxy group act.
• the aryl groups may be, for example, a phenyl, naphthalen-1-yl, naphthalen-2-yl, [1, 1'-biphenyl] -2-yl, [1, 1'-biphenyl] -3 -yl- or a [1, 1'-biphenyl] -4-yl group.
• the heteroaryl groups can be a pyridinyl, pyrimidinyl, quinolinyl, isoquinolinyl, benzofuranyl, benzothienyl, 1,3-benzodioxolyl, 2,1,3-benzothiadiazolyl, indolyl- , Furanyl, thienyl,
• the heterocyclyl groups for the radicals Z and Z ' may be a piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl, imidazolidinyl or pyrrolidinyl group linked via one of the substitutable sites.
• the substituents of the aryl, heteroaryl, heterocyclyl radicals may be, inter alia, unbranched or branched C 1 -C 4 -alkyl groups (methyl, ethyl, n-propyl, n-propyl, n-butyl, sec-butyl).
• Pent-4-enyl (E) -pent-3-enyl, (Z) -pent-3-enyl, (E) -pent-2-enyl, (Z) -pent-2-enyl- , 2-methylvinyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 2-methylbut-3-enyl, (E) -2-methylbut-2-enyl, (Z) -2-methylbut-2-enyl, 2-Ethylprop-2-enyl, Hex-5-enyl, (£) hex-4-enyl, (Z) -hex-4-enyl, (£) -hex-3-enyl, ( Z) -hex-S-enyl, (£) -hex-2-enyl, (Z) -hex-2-enyl, 1-methylpent-4-enyl, (E) -1-methylpent-3 -enyl-, (Z) -1-methyl
• C 3 -C 6 -cycloalkyl groups (cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl), and / or
• Halogen fluorine, chlorine, bromine, iodine
• Alkyl groups are identical or different, act.
• aryl-C 1 -alkylene groups for the radicals Z and Z ' may be a combination of the previously defined aryl and C 1 -C 4 -alkyl groups, for example: a
• the heteroaryl-C 1 -C 4 -alkylene groups for the radicals Z and Z ' may be a combination of the previously defined heteroaryl and dd-alkylene groups, for example a (pyridin-2-yl) methyl-, (pyridine) 3-yl) methyl, (pyridin-4-yl) methyl, (furan-2-yl) methyl, (furan-3-yl) methyl, (thien-2-yl) methyl, (thien) 3-yl) methyl, 2- (thien-2-yl) ethyl or a 2- (thien-3-yl) ethyl group.
• halogen to fluorine, chlorine, bromine or iodine.
• Preferred according to the present invention are those compounds of general formula I wherein
• Y is -OH, -CN, -SCN, a halogen atom, R 20 ;
• Y is -OH, -CN, -SCN, a halogen atom, R 20 ; or YR 20 , when Z and W together represent an oxygen atom,
• R 17 and R 17 ' together represent an oxygen atom, or
• R 17 is hydrogen, -OH
• R 17 represents hydrogen, -OH, -CC 4 -alkyl group, C 2 -C 5 -alkenyl group, a C 2 -C 5 - alkynyl group, or a trifluoromethyl group.
• the compounds according to the invention are suitable for inhibiting folliculogenesis and ovulation, for male contraception and for the treatment of benign and malignant proliferative disorders of the ovary.
• the compounds of the general formula I according to the invention alone, d. H. be used without the additional administration of progestins for contraception.
• ester derivatives of the estratrienes according to the invention can have as prodrug advantages over the unesterified active compounds with respect to their mode of application, their mode of action, potency and duration of action. Pharmacokinetic and pharmacodynamic advantages are also exhibited by the sulphamate derivatives of estratrienes according to the invention. These effects have already been described in other steroid sulfamates (Steroid Biochem Molec Biol 1995, 55, 395-403, Exp Opinion Invest Drugs 1998, 7, 575-589).
• the present invention describes 8 ⁇ -vinyl-11 ⁇ - (TO-substituted) alkyl-estra-1,3,5 (10) -trienes, the in vitro dissociation for binding to estrogen receptor preplarations of rat prostate and rat uterus, and those in vivo preferably have an inhibition of folliculogenesis and ovulation.
• the compounds of the invention have a contraceptive effect over a wide dose range, without other estrogen-sensitive organs, e.g. affecting the uterus or the liver.
• these compounds can be used for male contraception and for the treatment of benign or malignant proliferative diseases of the ovary.
• the present invention therefore relates to pharmaceutical preparations containing at least one compound of general formula I and their physiologically acceptable salts; the use of the compounds of general formula I for the manufacture of a medicament for male and / or female contraception, for the treatment of benign and malignant proliferative diseases of the ovary.
• the compounds according to the invention can be used for the following indications.
• the compounds of the general formula I according to the invention can be employed as individual components in pharmaceutical preparations or in combination, in particular, with GnRH antagonists, progesterone receptor antagonists, mesoprogestins, gestagens or tissue-selective gestagens (action via type A / B form).
• the compounds according to the invention and the preparations containing them are particularly suitable for ovarian contraception, for the treatment of benign or malignant proliferative diseases of the ovary, such as e.g. Ovarian carcinomas, granulosa cell tumors.
• the compounds may find use in the treatment of male fertility disorders and prostatic diseases.
• the amount of a compound of general formula I to be administered will vary within a wide range and may cover any effective amount. Depending on the condition to be treated and the mode of administration, the amount of compound administered may be 0.01 ⁇ g / kg-100 mg / kg body weight, preferably 0.04 ⁇ g / kg-1 mg / kg body weight, per day. In humans, this corresponds to a dose of 0.8 ⁇ g to 8 g, preferably 3.2 ⁇ g to 80 mg, r daily.
• a dosage unit according to the invention contains 1, 6 ⁇ g to 2000 mg of one or more compounds of general formula I.
• the compounds of the invention and their acid addition salts are suitable for the preparation of pharmaceutical compositions and preparations.
• the pharmaceutical compositions or medicaments contain as active ingredient one or more of the compounds according to the invention or their acid addition salts, optionally mixed with other pharmacologically or pharmaceutically active substances.
• the preparation of the drug is carried out in a known manner, wherein the known and customary pharmaceutical excipients and other conventional carriers and diluents can be used.
• carriers and excipients are, for example, those recommended or indicated in the following references as adjuvants for pharmacy, cosmetics and related fields: Ullmans Encyklopadie der ischen Chemie, Vol. 4 (1953), pages 1 to 39; Journal of Pharmaceutical Sciences, Vol. 52 (1963), page 918 et seq., H. v. Chr. Czetsch-ündenwald, adjuvants for pharmacy and adjacent areas; Pharm. Ind., Issue 2, 1961, page 72 u. ff. H. P. Fiedler, Lexicon of excipients for pharmacy, cosmetics and adjacent areas, Cantor KG. Aulendorf in WORK 1971.
• the compounds according to the invention can be administered orally or parenterally, for example intraperitoneally, intramuscularly, subcutaneously or percutaneously, or else be implanted in the tissue.
• the dosage units may contain, in addition to the active ingredient, a pharmaceutically acceptable carrier, such as starch, sugar, sorbitol, gelatin, lubricant,
• the active ingredients may be dissolved or suspended in a physiologically acceptable diluent.
• oils are often used with or without the addition of a solubilizer, surfactant, suspending or emulsifying agent. Examples of oils used are olive oil, peanut oil, cottonseed oil, soybean oil, castor oil and sesame oil.
• the compounds may also be used in the form of a depot injection or an implant preparation, which may be formulated to allow sustained release of active ingredient.
• Implants may contain as inert materials, for example, biodegradable polymers or synthetic silicones such as silicone rubber.
• the active ingredients can also be incorporated for percutaneous administration, for example in a plaster.
• intravaginal e.g., vaginal rings
• intrauterine systems e.g., pessaries, spirals, lUSs
• various polymers such as silicone polymers, ethylene vinyl acetate, polyethylene or polypropylene are suitable.
• the compounds may also be formulated as cyclodextrin clathrates.
• the compounds with ⁇ -, ß- or ⁇ -cyclodextrin or derivatives of these are implemented (PCT / EP95 / 02656).
• the compounds of the general formula I according to the invention can also be encapsulated with liposomes.
• the binding affinity of the compounds of the invention was tested in competition experiments using 3H-estradiol as a ligand on estrogen receptor preparations of rat prostates and rat uteri.
• the preparation of the prostate cytosol and the estrogen receptor test with the prostate cytosol was performed as described by J. Testas et al. in Endocrinology 1981, 109, 1287-1289.
• the preparation of rat uterus cytosol, as well as the receptor assay with the ER-containing cytosol were carried out in principle as described by Stack and Gorski in Endocrinology 1985, 117, 2024-2032, with some modifications according to U. Fuhrmann et al. in Contraception 1995, 51, 45-52).
• the compounds of the invention have higher binding affinity for estrogen receptor from rat prostate than for estrogen receptor from rat uterus (Tables 1 and 2). It is assumed that ER ⁇ predominates over ER ⁇ in the rat prostate and in rat uterus ER ⁇ over ER ⁇ . Table 1 shows that the ratio of prostate and uterine receptor binding qualitatively agrees with the relative binding affinity (RBA) ratio of rat human ER ⁇ and rat ER ⁇ (according to Kuiper et al., Endocrinology 1996, 138, 863-870) (Table 1) ).
• RBA relative binding affinity
• U-2 OS cells are grown in Dulbecco 's medium (DMEM) without phenol red (Gibco BRL; # 11880-028) + 5% fetal calf serum (FKS) (Seromed; # S 0115) + 100 units / ml penicillin / 100 ⁇ g / ml Streptomycin (Seromed, #A 2213), 4 mM L-glutamine (Gibco BRL; # 25030-024) (PSG) was cultured at 37 ° C and 8.5% CO 2 .
• DMEM Dulbecco 's medium
• FKS fetal calf serum
• PSG 4 mM L-glutamine
• the ER ⁇ expression plasmid (HEGO) used was amplified in E. coli DH5 ⁇ (from Invitrogen).
• the ER ⁇ expression plasmid (ER ⁇ O) used was produced in house and amplified in E. coli DH5 ⁇ .
• the expression plasmid pSG5 was used.
• the vector pBL-LUC + was provided with two tandem EREs (estrogen-responsive elements of the vitellogenin promoter) and amplified in E. coli (XL1-Blue, Fa. Stratagene).
• Plasmid DNA is prepared using the NucleoBond Plasmid Maxi Kit (CLONTECH; # K3003-2) and FuGENE 6 Reagent (Boehringer Mannheim, # 1 814 443). These are first diluted separately in a suitable volume of DMEM and incubated before combining the solutions and incubating again. Approaches for 96-well plates:
• the dilution stages are chosen such that the final concentrations on the test plate for agonism in the range of 10 - 12 M "7 10" (for E2: 10 -8 - 10 _13 M) lie.
• the transfection medium is replaced by 180 ⁇ l D-MEM + 5% CCS +
• the cells are additionally treated with estradiol. Subsequently, 20 ⁇ l of the substance dilutions are added by pipette. The negative controls receive 20 ⁇ l DMEM + 1% DMSO per well. The final test substance concentrations are 3x10 _1 M for ER ⁇ and 3x10 _ 0 M for ER ⁇ , respectively.
• the reference substance used is the known antiestrogen fulvestrant (AstraZeneca) in the same concentrations (Table 2).
• the lysates are mixed with 30 ⁇ l of luciferase substrate A (PharMingen, # 556867) and 30 ⁇ l of luciferase substrate B (PharMingen, # 556869).
• the measurement of the luciferase activity takes place 30 seconds after the addition of substrate B in the cycle mode of
• Luminometer (DYNATECH, ML3000).
• the evaluation of the measured data takes place by means of the device manufacturer
• Immature female rats are hypophysectomized. This tag is defined as day 0. From Day 1 - Day 4, treatment, subcutaneous and / or oral, is with the active substance in combination with 17 ⁇ -oestradiol. On day 5, a subcutaneous injection with PMSG (pregnant mare serum gonadotropin) is performed. On day 7, hCG is administered intraperitoneally to induce ovulation. On day 8, the ovary is harvested and analyzed macroscopically (e.g., ovary weights) and / or microscopically (e.g., follicular histological evaluation, so-called follicle staging). The tubes are rinsed and examined for the presence of egg cells.
• PMSG pregnant mare serum gonadotropin
• Immature female rats are subcutaneously treated with PMSG (pregnant mare serum gonadotropin) at day 23 (day 1).
• PMSG pregnant mare serum gonadotropin
• the animals receive the active substance administered subcutaneously or orally.
• the animals are given an intraperitoneal injection of hCG to induce ovulation.
• Autopsy is performed 16 hours after hCG administration. The tubes are rinsed and examined for the presence of oocytes.
• Another possibility for detecting the dissociated estrogen effect of the substances according to the invention in vivo is, after single application of the substances in rats, to measure effects on the expression of 5HT2a receptor and serotonin transporter protein and mRNA levels in ER ⁇ -rich brain areas. Compared to the effect on serotonin receptor and transporter expression, the effect on LH secretion is measured. Substances with higher binding to the rat prostate compared to the rat uterine estrogen receptor are more potent as regards Increasing the expression of serotonin receptor and transporter, compared to their positive effect on LH secretion. The density of serotonin receptor and transporter is determined by radioactive ligands, and the corresponding mRNA by in situ hybridization. The method is described in the literature: G. Fink & BEH Sumner 1996 Nature 383: 306; BEH Sumner et al. 1999 Molecular Brain Research, in press.
• the present invention also relates to the intermediates of general formula VI
• the present invention further relates to processes for the preparation of the compounds of general formula I, and in each case a process for the preparation of the individual intermediates VI to IX.
• the compounds of general formula I according to the invention can be prepared as described in the examples. By analogous procedure using homologous reagents to the reagents described in the examples, the other compounds of general formula I can be obtained.
• Etherification and / or esterification of free hydroxy groups takes place by methods familiar to the person skilled in the art.
• the compounds according to the invention can be present at the carbon atom 17 as ⁇ , ⁇ -stereoisomers.
• the compounds are usually obtained as mixtures of the corresponding ⁇ , ß-isomers.
• the mixtures can be separated, for example, by chromatographic methods.
• Substituents which are possible according to the general formula I may already be present in the starting form or in the form of a precursor in the starting product, an estron already corresponding to the desired end product.
• 17-Substituents are, also by known methods, introduced by nucleophilic addition of the desired substituent or a reactive precursor thereof, and optionally further developed.
• estratriene carboxylic acid esters according to the invention are prepared analogously to likewise known processes from the corresponding hydroxysteroids (see, for example, Pharmaceutical Active Ingredients, Syntheses, Patents, Applications, A. Kleemann, J. Engel, Georg Thieme Verlag, Stuttgart 1978. Arzneistoff, Fort Kunststoff 1972 to 1985 A. Kleemann, E. Lindner, J. Engel, VCH 1987, pp. 773-814).
• estratriene sulfamates according to the invention are obtainable in a manner known per se from the corresponding hydroxy steroids by esterification with sulfamoyl chlorides in the presence of a base (Z. Chem., 1975, 15, 270-272, Steroids 1996, 61, 710-717). Subsequent acylation of the sulfamide group leads to the (N-acyl) sulfamates according to the invention, for which pharmacokinetic advantages have already been demonstrated in the absence of an 8-substituent (compare WO 97/14712).
• the preparation of the sulfamates according to the invention with one or more additional hydroxyl groups in the molecule is also possible by starting from suitable hydroxy-steroid ketones.
• one or more hydroxyl groups are subjected to sulfamoylation.
• the sulfamate groups can optionally be converted with a desired acyl chloride in the presence of a base in the respective / N-acyl) sulfamates.
• the now present oxosulfamates or oxo (N-acyl) sulfamates are converted by reduction into the corresponding hydroxysulfamates or hydroxy- (N-acyl) sulfamates (Steroids 1996, 61, 710-717).
• Suitable reducing agents are sodium borohydride and the borane-dimethyl sulfide complex.
• substituents according to the general formula I can also be introduced at the stage of the estratrienes already substituted in the 8-position. This can be useful or necessary in particular in the case of multiple substitution of the desired end compound.
• 1, 3,5 (10), 9 (11) -tetraenes can be, as well as the 8ß-substituted 11 ß-alkyl estraene
• THP tetrahydropyran-2-yl
• Tf trifluoromethanesulfonyl
• TBS tert-butyldimethylsilyl
• a solution of 28 ml of diisobutylaluminum hydride in 83 ml of toluene is added dropwise at -10 ° C. to a solution of 15.4 g of nitrile 4 in 280 ml of toluene.
• the reaction solution is stirred until complete reaction at 0 ° C, successively with 460 ml of toluene, 92 ml of sat.
• Sodium bicarbonate solution and 9 ml of 2-propanol stirred for several hours at room temperature. It is then filtered through Celite and the filtrate is concentrated.
• the colorless foam thus obtained is dissolved in 280 ml of ethanol / water (5: 1), 28.75 g of p-toluenesulfonic acid are added, and the reaction solution is heated to 60 ° C. and stirred until complete reaction. Subsequently, a large portion of the ethanol is removed on a rotary evaporator, the residue diluted with ethyl acetate, washed with water, sat. Sodium bicarbonate and sat. Sodium chloride solution, dried over magnesium sulfate and concentrated.
• a solution of 1 ml of diisobutylaluminum hydride in 1 ml of toluene is added dropwise at -10 ° C. to a solution of 1.16 g of cyanide 11a in 23 ml of toluene.
• the reaction solution is stirred until complete reaction at -10 ° C, successively with 17 ml of toluene, 6 ml of sat.
• Sodium bicarbonate solution and 0.7 ml of 2-propanol stirred for several hours at room temperature. It is then filtered through Celite and the filtrate is concentrated.
• a solution of 0.4 ml of diisobutylaluminum hydride in 0.4 ml of toluene is added dropwise at -10 ° C. to a solution of 480 mg of cyanide 11b in 10 ml of toluene.
• the reaction solution is stirred until complete reaction at -10 ° C, successively with 8 ml of toluene, 3 ml of sat.
• Sodium bicarbonate solution and 0.3 ml of 2-propanol stirred for several hours at room temperature. It is then filtered through Celite and the filtrate is concentrated.
• the resulting trimethylsilyl ether proves to be partially unstable during subsequent purification by column chromatography and is reacted in a mixture with its corresponding alcohol.
• the mixture is dissolved in tetrahydrofuran (10 ml / mmol), treated at room temperature with tetrabutylammonium fluoride trihydrate (1.5 equiv.) And stirred until complete reaction at room temperature.
• tetrahydrofuran (10 ml / mmol)
• tetrabutylammonium fluoride trihydrate 1.5 equiv.
• For workup is diluted with diethyl ether, the organic phase with water and sat. Sodium chloride solution, dried over magnesium sulfate and concentrated.
• Column chromatographic purification (cyclohexane / ethyl acetate) gives the corresponding perfluoroalkyl-substituted alcohols as colorless foams.
• estratriendiol 18a as a colorless foam (GC-MS: m / z theor .: 520, practical: 520) together with 46 mg of the by-produced unreacted trimethylsilyl ether, which by reaction with Tetrabutylammonium fluoride trihydrate in tetrahydrofuran (see procedure 1.1 / trimethylsilyl ether cleavage) is converted into compound 18a (33 mg).
• steroid 17c give, in the reaction analogous to procedure 1.2, 57 mg of the unreacted trimethylsilyl ether which, in the reaction with tetrabutylammonium fluoride trihydrate in tetrahydrofuran (cf., regulation 1.1 / trimethylsilyl ether cleavage), contains 45 mg of estratriene diol 18c as a colorless foam (GC-MS: m / z theor .: 548, practical: 548).
• the colorless foam obtained is dissolved in 1 ml of toluene, added dropwise at 0 ° C 0.15 ml of diisobutylaluminum hydride and the reaction solution heated to reflux until complete. After cooling to 0 ° C, the reaction mixture is successively with 0.5 ml of ethanol, 0.5 ml of ethanol / water (1: 1) and 0.5 ml semicon. Hydrochloric acid and stirred for about 30 minutes. The phase separation takes place between diethyl ether / water. The organic phase is washed with sat. Sodium chloride solution, dried over magnesium sulfate and concentrated.
• the organic phase is washed with water and sat. Washed sodium chloride solution, dried over magnesium sulfate and concentrated.
• the column-chromatographic purification is carried out on silica gel with a cyclohexane / ethyl acetate mixture as eluent and gives the corresponding carboxylic acids.

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