JPH09510959A - Novel antiandrogens and related pharmaceutical compositions and methods of use - Google Patents

Abstract

(57) [Summary] A novel anti-androgen agent is provided. Exemplary groups of compounds have the structural formula (I), wherein R ¹ to R ^10, a and b are as defined herein. Provided are pharmaceutical compositions and methods of using the compounds of formula (I) for treating androgen-related clinical conditions, as well as methods and compositions of using the compounds as contraceptives.

Description

Detailed Description of the Invention       Novel antiandrogens and related pharmaceutical compositions and methods of use   Government endorsement   This invention was partially funded by the National Institutes of Health under Approval No. AM33747-03. Provided; therefore, the US Government may have certain rights in this invention.   Technical field   The present invention relates generally to steroid hormones, and more specifically to antiandrogens And useful novel compounds. Further, the present invention is androgen responsive Treatment of clinical conditions that are either associated with or androgen excess And a pharmaceutical composition for treating such a clinical condition. Sa Moreover, the present invention relates to the use of the novel compounds as male contraceptives.   background   Antiandrogens are androgen-responsive or are androgen-rich Clinical conditions that are either related to (eg, prostate carcinoma, good Prostatic hyperplasia (BPH), acne, seborrhea, alopecia, hirsutism, polycystic ovarian disease, And male pattern baldness). Antiandrogen treatment , May relate to any regulatory step in androgen production or androgen action . Regulation of androgen production in the testis is controlled by the hypothalamic-pituitary peptide hormone. Is mediated directly by. Hypothalamic neurons are gonadotropin-releasing High-affinity cell surface receptor on the plasma membrane of pituitary cells that secretes monone (LHRH) Secrete a decapeptide that interacts with the site. LHRH is luteinizing hormone (LH) Release of both follicle-stimulating hormone and follicle-stimulating hormone (FSH) by a calcium-dependent mechanism. Stimulate.   LH secretion is related to the reaction of androgens and estrogens in the hypothalamus and pituitary gland. Therefore, it is controlled. The control of LH in men is primarily due to negative feedback. Happens. Because testicular steroids inhibit LH secretion. test Both sterone and estradiol can inhibit LH secretion. Testostero Can be converted to estradiol in the brain and pituitary, but two hormones Probably works independently. Testosterone acts on the central nervous system (CNS) Slow down floor pulse generator, resulting in reduced frequency of LH pulsed secretion . In addition, testosterone has a negative feedback on LH secretion in the pituitary gland. It seems to do.   LH reaches the testis by peripheral circulation, where it is the plasma membrane of Leydig cells. Interacts with certain high affinity cell surface receptors above. LH to LH receptor Binding stimulates testosterone biosynthesis.   Testosterone, the major secreted product, is dihydrotestosterone (DHT ), Androsterone, androstenedione, progesterone, and 17-hydr Roxyprogesterone is also secreted by the testes.   In peripheral tissues, testosterone (eg CNS, skeletal muscle and seminiferous epithelium Formation of DHT as a circulating prohormone (eg. For example, prostate) and estrogen formation. In the prostate , Testosterone is diffused into cells, where testosterone is 5-alpha-red. Reduced to DHT by cutase. 90% of total prostate androgen is in the form of DHT It is mainly derived from testis androgen. Prostate androgen rest 10% of this is produced in the adrenal glands. In the cells of the prostate, testostero And DHT bind to the same high affinity androgen receptor protein I do. The hormone receptor complex then binds to specific DNA in the nucleus of prostate cells. It binds to the binding site. This results in increased transcription of androgen-dependent genes and Which ultimately results in stimulation of protein synthesis. Conversely, from androgen-sensitive tissues Withdrawal of androgens reduces protein synthesis, tissue decay, and some In the case of, cells die.   As mentioned above, many physiological problems are associated with androgen production. At the moment The most pressing of these is prostate cancer, which in humans Is the leading cause of cancer in cancer, diagnosed in 100,000 orders per year, and 26, 000 people die. Androgen dependence of some prostate cancers is well established Androgen is the leading treatment for metastatic prostate cancer. Suppression. Androgen suppression can be accomplished as follows: sperm Removal of the testes, a major source of androgens, by nestectomy; luteinizing hormone Pituitary levels by either mon-releasing hormone analogs or estrogens Of testicular steroidogenesis in humans; testis-level sperm using enzyme inhibitors Inhibition of focal steroid production; or by androgen receptor antagonists Inhibition of the action of androgens.   Another important therapeutic area related to androgen production and action is in the United States alone. Benign prostatic hyperplasia, i.e. an old man with approximately 400,000 prostatectomy performed between Cure of major sexual problems (and prostate cancer precursors in about half of diagnosed cases) It is a cure. Surgical medicine currently represents the most established treatment for BPH, but Several pharmacological approaches have been tested as well. But still, drug treatment Is not satisfied.   Known antiandrogens act by several mechanisms. Pituitary gland There are drugs that inhibit LH secretion and reduce testosterone and DHT production; They are called "LHRH agonists" and, for example, nafarelin, leupro Includes Lido, Goserelin and Buserelin. Similarly, it inhibits pituitary LH secretion, Reduces the production of testosterone and DHT, but also the androgen receptor -There are additional agents that also inhibit; these are cyproterone acetate, zanotero Logestin (eg megestrol acetate, hydroxy-progesterone Drugs such as caproate, and medrogestone, and negative feedback It exhibits anti-androgenic effects through the process. Other antiandrogens are non-sulfur Androgen receptor inhibitor) and 5-α-reductase inhibitor Includes bitters (eg finasteride), which reduce DHT selectivity .   However, currently available antiandrogens are associated with widespread problems. For example, public Known antiandrogens have many side effects, such as impotence, decreased sexual drive, females, among others. It causes sexualized breasts, heat intolerance, and hot flashes. Several The drug is associated with fatal hepatotoxicity (eg, D.K.Wysowski et al.Ann. Int. Med.  118(11): 860-864 (1993)). Moreover, known drugs have a very short half-life. And require more frequent and / or higher doses.   The present invention relates to the above need in the art and to certain novel 17-positions. The substitution steroids are associated with prior art antiandrogen compounds at relatively low doses. Presumed to be useful as an antiandrogen without causing problems And Therefore, the new compound may cause the above-mentioned undesirable side effects. Non-androgen responsive and / or associated with androgen production Useful for treating clinical conditions. Therefore, this compound Chinoma, benign prostatic hyperplasia, acne, seborrhea, alopecia, hirsutism, polycystic ovarian disease It is useful for treating patients and male pattern baldness. In addition, this compound is a male contraceptive Is also useful as While not wishing to be bound by theory, the present inventors Most of these new drugs are stimulated by exogenous testosterone. By inhibiting the growth of the prostatic Acts as a gonist, and is essentially a progesten and agonist andro Considered to lack gen activity.   Background technology   The following documents relate to known antiandrogen compounds and / or molecular structures, These are related in some respects to the novel agents described herein: Anner et al. U.S. Pat. No. 4,150,127 (19-oxygenated supra described as useful as a diuretic) Pyroxane-type steroids); Sokolowski U.S. Pat. No. 4,41 2,993 (in the treatment of pseudopregnancy, galactorrhea, and mastitis in mammals Describes 7α-methyltestosterone said to be useful); Lau rent et al., U.S. Pat. No. 4,673,673 (17α-alkyl-17 as an antiandrogen β-hydroxy-1α-methyl-4-androsten-3-one is described); Nique U.S. Pat. No. 4,874,754 (as a contraceptive or for the treatment of certain gynecological disorders. Specific 19-nor steroids for Xu No. 4,891,365 (specific 17-substituted estradiene for the treatment of gynecological diseases and And Estratriene are listed). Wiechert et al. U.S. Pat.No. 4,456,600, Nos. 4,558,041 and 4,892,867 all describe 17α-substituted steroids. All of these patents are useful for treating acne, seborrhea, alopecia, hirsutism. Anti-androgens. In addition, G. Ohta et al. the study. IV. Synthesis of Androstano [2,3-c] -furazane and Related Compounds, Chem. Pharm. Bull.13: 1445-1459 (1965), and Ferrari et al., “Three species of laboratory animals Endocrine Profile of Topterone, a Topical Antiandrogen, "Meth and F ind Exptl Clin Pharmacol2(2): 65-69 (1980) relates to related steroid structures. The purpose is to   Summary of the Invention   Therefore, the main object of the present invention is to identify new compounds useful as antiandrogens. By providing, it addresses the above-mentioned need in the art. .   Another object of the present invention is androgen responsiveness or androgen excess. Providing a pharmaceutical composition for treating a clinical condition that is either associated with Is to provide.   Yet another object of the invention is a pharmaceutical composition for use as a male contraceptive. Is to provide.   Yet another object of the present invention is androgen responsiveness or androgen To provide a method of treating a clinical condition that is either associated with And.   A further object of the present invention is to provide an effective androgen dose in combination with a compound as described above. It is intended to provide a method which comprises the oral administration of a product.   Another further object of the present invention is to provide a fertile male within a predetermined dosing regimen. By providing a contraceptive method of orally administering a compound to a mammal of the above. is there.   Further objects, advantages and novel features of the present invention will be explained in part in the description below. And shown in part to one of ordinary skill in the art with regard to the following description. Or can be understood by practice of the invention.   In one embodiment, the present invention provides an antiandrogen and / or contraceptive agent. And within the effective dosing regimen to achieve the desired intended result, Regarding methods of using the compound having formula (I): In formula (I):   R¹And R²Are independently selected from the group consisting of hydrogen and lower alkyl, And R^ThreeAnd R^FourAre independently hydrogen, hydroxyl, and lower alkoxy? Selected from the group consisting of or R^ThreeAnd R^FourTogether with one carbonyl group Where R^ThreeAnd R^FourOne of is hydroxyl or lower alkoxy When the other is hydrogen;   Or R¹And R²One is R^ThreeAnd R^FourOne of the following rings Provides:   R if "a" does not represent a double bond^FiveIs hydrogen or lower alkyl, and "a R represents a double bond^FiveNot;   R⁶And R⁷Are independently hydrogen, hydroxyl, and lower alkyl. Selected from the group⁶And R⁷Together represent one carbonyl group Or a cycloalkyl ring containing 3 to 6 carbon atoms, which may be attached. Or may provide a cyclooxyalkyl ring;   R⁸And R⁹Are independently selected from the group consisting of hydrogen and lower alkyl;   R if "b" does not represent a double bond^TenIs hydrogen and “b” represents a double bond. If you^TenNot; and   a and b represent a double bond as needed.   The present invention also provides a pharmaceutical composition containing one or more compounds of structural formula (I) With respect to the compounds, and as further detailed below, is included in formula (I) Particular novel compound.Brief description of the drawings   FIG. 1 is a graph illustrating the effect of drug administration on prostate weight. This will be described in Example 3.Detailed description of the invention Definition and naming   Prior to disclosing and describing the compounds, compositions and methods of the present invention, the present invention Specific reagents or reaction conditions, specific pharmaceutical carriers, or specific administration methods It is to be understood that the present invention is not limited to the above, and can naturally change. Used herein The term is intended to describe only particular embodiments and is not intended to be limiting. Should be understood.   As used herein and in the appended claims, the singular form "a", "An" and "the" are in the plural unless the context clearly dictates otherwise. It should be noted that it includes the referent. So, for example, "Anti Andro Reference to "genetic agent" includes a mixture of antiandrogens and refers to "pharmaceutical carrier". Reference to "a" includes a mixture of two or more such carriers. And so on.   In this specification and in the claims that follow, it is defined as having the following meanings: Many terms to be mentioned are mentioned:   As used herein, the term "alkyl" has 1 to 24 carbon atoms. , Branched or unbranched saturated hydrocarbon groups, such as methyl, ethyl, n-pro Pill, isopropyl, n-butyl, isobutyl, t-butyl, octyl, decyl, It refers to tradecyl, hexadecyl, icosyl, tetracosyl, etc. In this specification A preferred alkyl group in the group contains from 1 to 12 carbon atoms. The term "lower al “Kil” means an alkyl group having 1 to 6, preferably 1 to 4 carbon atoms. Intended. The term "cycloalkyl" typically refers to 3 to 6 carbon atoms, More preferably, a cyclic alkyl group having 4 to 5 carbon atoms is intended. the term" Cyclooxyalkyl "contains one ether linkage and is more typically 3 Cyclic alkanes containing 1 to 6 carbon atoms, more preferably 4 to 5 carbon atoms. Intends a kill group.   The term "alkenylene" has 2 to 24 carbon atoms and at least one double Refers to a bifunctional branched or unbranched hydrocarbon chain containing a bond. "Lower Arche “Nylene” is an alkene having from 2 to 6, more preferably 2 to 5 carbon atoms. Refers to a nylene group.   The term "aryl" as used herein is a monocyclic ring of 5 to 7 carbon atoms. An aromatic species, typically phenyl. If desired, these groups are 1 One to four, more preferably one to two lower alkyl, lower alkoxy, hydro Substituted with xy and / or nitro substituents.   “Halo” or “halogen” refers to fluorine, chlorine, bromine, or iodine, Usually, it means a halo substitution product of a hydrogen atom in an organic compound. Of the halo, chlorine and And fluorine are generally preferred.   “As needed” or “as needed” means an event or Facts may or may not occur, It is meant to include when and when fruit does not occur. For example, the phrase "Double bond optionally present" means that a double bond may or may not be present. Both the case where the double bond is present and the case where it is not present. It means to include the person. The dashed line adjacent to the solid line is "double bond if necessary. , And thus a double bond may or may not be present (double bond If no bond exists, adjacent atoms are covalently bonded by a single bond). means.   The term "effective amount" or "antiandrogen effective" of the agents provided herein "Amount" is a nontoxic but sufficient amount of drug to provide the desired antiandrogenic effect. Means the amount of. As noted below, the exact amount required will depend on the species of the subject. Type, age, and general condition, severity of condition to be treated, and specific It varies depending on the subject depending on the anti-androgen agent and its administration form. Therefore, it is not possible to specify an exact "effective amount". But a valid effective The amount can be determined by one of ordinary skill in the art using only routine experience.   "Pharmaceutically acceptable" means a non-biological or other unwanted substance That is, the substance causes any undesired biological effects Without or deleteriously interact with any other ingredients of the pharmaceutical composition containing the substance. It can be administered individually with the selected anti-androgen agent without use.   When describing the positions of groups and substituents, the following numbering system is used:   This system uses the number of cyclopentanophenanthrene nuclei as IUPAC or Chemical Ab Intended to follow the conventions used by the stracts Service. In this specification The term "steroid" as used has the cyclopentanophenanthrene nucleus described above. Is intended to mean a compound.   In addition, in these structures, bold lines and lines showing the particular conformation of the group And dashed lines follow the IUPAC Steroid Nomenclature Convention. The symbols “α” and “β” are drawn The specific stereochemical configuration of substituents on asymmetric carbon atoms in such a chemical structure Show. In other words, the “α,” indicated by the broken line is the group in question. It is shown below the general plane of the molecule and is indicated by a thick line. The “β,” that appears is that the group at the problematic position is on the paper surface of the molecule as drawn. Indicates that   Furthermore, the 5- and 6-membered rings of steroid molecules are often, as shown, Called A, B, C and D.New compound :   The novel compounds provided herein are defined by structural formula (I) as described above. A compound, which is defined as R¹~ R^Ten, A and b are as described herein above. As defined in, except that (1) R¹And R²One side is R^ThreeAnd R^FourOne of When connecting to Provides R⁶And R⁷Are not hydroxyl; and (2) R^ThreeAnd And R^FourWhen together represent one carbonyl group, R^FiveIs hydrogen, there is no "a" . Preferred compounds included in formula (I) are included in the following two groups.   A first group of compounds can be represented by the following structural formula (II): In this subset of compounds, R¹And R²Are independently hydrogen and lower Selected from the group consisting of alkyl, and R⁶, R⁷, R⁸, And R⁹In formula (I) As defined above. Particularly preferred compounds in this subset Has the following structural formula (II-1):   A second group of compounds may be represented by the following structural formula (III): In a subset of this compound, R^FiveIs either hydrogen or lower alkyl And then R⁶, R⁷, R⁸, And R⁹Is as defined above for formula (I) is there. Examples of particularly preferred compounds in this subset are represented by the following structural formula (III-1) Having:   Other examples of compounds included in structural formula (I) are as follows:   Uses and administration:   The compounds defined by structural formula (I) are useful as antiandrogens , Therefore prostate carcinoma, benign prostatic hyperplasia, acne, seborrhea, alopecia, With androgen responsiveness such as hirsutism, polycystic ovarian disease, and male pattern baldness Useful for the treatment of clinical conditions that are present and / or associated with androgen excess It is. The compound is combined with one or more pharmaceutically acceptable carriers. Can be successfully formulated into a pharmaceutical composition composed of a compound ofRemington's Ph armaceutical Science The latest edition, edited by E.W.Martin (Mack Publ. Co., Easton PA) , Formulations using typical carriers and anti-androgen compounds of the present invention. It discloses conventional methods of preparing pharmaceutical compositions that can be used to make. This compound may also be administered in the form of pharmacologically acceptable salts or esters. You. Salts or esters of this compound are standard salts known to those skilled in synthetic organic chemistry. Can be prepared using the procedure described in J. March,Advanced Organic Chemistry: React ions, Mechanisms and Structure 4th Edition (New York: Wiley-Interscience, 1992) )It is described in. This disclosure is hereby incorporated by reference herein. Used. The ester was prepared by using C-17 hydroxyl group as a functional group of steroid molecule. (Eg, introduction of succinate, malonate, glutarate groups, etc.) Including.   Is the compound administered orally or parenterally (eg, intravenously) , Topically, transdermally, by intramuscular injection, and Is administered by intraperitoneal injection or the like, but oral administration is preferred. Activity administered The amount of the compound will, of course, be the subject to be treated, the weight of the subject, the mode of administration and the formulation. To It depends on the judgment of the doctor. However, in general, the dosage is about 0.5 mg / kg / day to 20 mg / kg / day. , And more typically about 1.0 mg / kg / day to 10 mg / kg / day.   Depending on the intended form of administration, the pharmaceutical composition may be a solid, semi-solid or liquid dosage. In a given form (eg tablets, suppositories, pills, capsules, powders, solutions, suspensions, etc.) Possible, preferably in unit dose form suitable for single administration of the correct dose . The composition is selected in combination with a pharmaceutically acceptable carrier, as described above. The drug contains an effective amount of Excipients and the like.   For solid compositions, conventional non-toxic solid carriers include, for example, medicated gray. De mannitol, lactose, starch, magnesium stearate, sacchar Sodium phosphate, talc, cellulose, glucose, sucrose, magnesia carbonate Including um. Liquid pharmaceutically administrable compositions are described herein, for example. And an optional pharmaceutical adjuvant in an excipient (Eg water, saline, dextrose solution, glycerol, ethanol Solution, suspension, etc., thereby preparing a solution or suspension Form a liquid. If desired, the pharmaceutical composition to be administered may also contain traces of wetting. Non-toxic auxiliary substances such as agents or emulsifiers, pH buffering agents (eg sodium acetate) Um, sorbitan monolaurate, triethanolamine sodium acetate, tri Ethanolamine oleate, etc.). To prepare such a dosage form The actual methods of doing so are known or apparent to those skilled in the art; aboveRemington's Pharmaceutical Sciencereference.   For oral administration, finely divided powders or granules may be used as diluents, dispersions and / or capsules. May contain a surface active agent and may be present in water or in a syrup or in the dry state May be present in capsules or sachets, or contain suspending agents May be present in a non-aqueous solution or suspension, or may be binders and lubricants May be present in tablets, or may be in suspension in water or syrup. Can exist. Fragrances, preservatives, suspending agents, if desired or necessary Thickeners or emulsifiers may be included. Tablets and granules are preferred oral dosage forms And these can be coated.   Parenteral administration, if used, is generally characterized by injection. Injection Quality may be in conventional form, in liquid solution or suspension form or in liquid prior to injection. Either as a solid form suitable for solution or suspension, or as an emulsion Can be prepared. A recently modified approach to parenteral administration has been Includes the use of slow or sustained release systems to maintain dosage. For example , U.S. Pat. No. 3,710,795. This is incorporated herein by reference.Preparation process   The compounds of the present invention have high yields, as exemplified in the experimental section herein. At a rate, it can be prepared using a relatively simple direct method.   The synthesis of representative compounds is detailed in the examples below. Compound II-1 was synthesized The synthesis of compound III-1 is described in detail in Example 2, while described in detail in Example 1. It is.   Biological test procedures and results are described in Example 3 below.Experiment   The practice of the present invention, unless otherwise indicated, includes synthetic organic chemistry, biological test methods, etc. Conventional techniques are used. These are within the purview of those skilled in the art. this Such techniques are explained fully in the literature. For example, steroid-related synthetic procedures For more information on Fieser et al.,Steroids(New York: Reinhold, 195 9), and Djerassi,Steroid Reactions: An Outline for Organic Chemists(S an Francisco: Holden-Day, 1963). Described herein, and Steroids useful for assessing compounds as claimed in For more information on the Lumon Receptor Competition Assay,J. Steroid Biochem. 12: 143 See -157 (1980). Mentioned herein, both above and below. All patents, patent applications, and publications cited are incorporated herein by reference. You.   While the present invention has been described with respect to certain preferred embodiments thereof, the above description The following and the following examples are intended to illustrate and limit the scope of the invention. Not intended.   Other aspects, advantages and modifications within the scope of the invention will occur to those skilled in the art to which the invention pertains. Is obvious.   In the examples below, the numbers (eg, amounts, temperatures, etc.) used in the examples Efforts have been made to ensure accuracy but some experimental errors and deviations should be accounted for. Should be. Unless stated otherwise, temperature is in degrees Celsius (° C) and pressure is atmospheric pressure. Or near atmospheric pressure. All solvents were purchased as HPLC grade and all All reactions were routinely run under an inert atmosphere of argon unless otherwise noted. Was. NMR analysis was performed on either a Varian XL-400 or JEOL FX90Q and a δ7.27 With chloroform as the reference. FTIR spectra recorded on Perkin-Elmer 1610 .                                   Example 1   This example shows that as shown in Scheme 1, 17α-propyl estra-4,9-diene-1. The synthesis of 7β-ol-3-one (II-1) is described.   (a.) 3-methoxy-17α-propynyl estra-1,3,5-trien-17β-ol (2) :   3-methoxy estrone (19.02 g, 31.8 mmol) in THF (250 ml), A solution of Nylmagnesium bromide (120 mL of 0.56M in THF) was added. Got The yellow solution was heated under reflux for 3 hours, then cooled to room temperature and saturated NH 4_FourCl water soluble Stir with liquid (100 mL) and extract with ethyl acetate (3 x 100 mL). one The combined organic layers were washed with brine (2 x 100 mL) and MgSO_FourAnd then dry Porated to give a yellow solid. From a mixture of ethyl acetate and hexane Recrystallization was performed to obtain white fine crystals (8.77 g (85%), melting point 199 ° C to 200 ° C). HRMS. C_{twenty two}H₂₈O₂Calculated for: 324.209. Found: 324.210.   (b.) 3-methoxy-17α-propyl estra-1,3,5-trien-17β-ol (3) :   Alkyne2(4.00 g, 12.3 mmol), 5% Pd / C (100 mg), and ethyl acetate (4 00mL) suspension in H₂The mixture was stirred for 14 hours under the atmosphere. The catalyst is removed by filtration, The filtrate was concentrated under vacuum to give a white foam. In the subsequent process (in successive crops ) From methanol to give 3.70 g (92%) of white needles (melting point 96 ° C-97 ° C). Obtained. HRMS. C_{twenty two}H₃₂O₂Calculated value for: 328.240. Found: 328.240.   (c.) 3-methoxy-17α-propyl estra-2,5 (10) -dien-17β-ol (4) :   Lithium particles (0.63 g) were added to liquid ammonia (350 mL) at -78 ° C. Metal melts Then dry isopropanol (30 mL) and aromatic3(4.70 g, 14.3 mmol) THF solution (100 ml) was continuously added. After 30 minutes at -78 ° C, wash the resulting blue solution. Warm to room temperature overnight. The resulting cloudy solution was washed with saturated aqueous ammonium chloride solution (2 Stirred with 00 mL) and diethyl ether (200 mL). Separate the ether layer and get tired Wash with aqueous ammonium chloride solution (100 mL) and brine (2 x 100 mL), K₂CO_Three Dried in vacuo and evaporated to give a white amorphous solid (4.90, 100%). this Was used without further purification, and diethyl ether and hexane Crystallization from a mixture of the above gave purified crystals for analysis (melting point 148 ° C to 149 ° C). HRMS. C_{twenty two}H₃₄O₂Calculated for: 330.256. Found: 330.256.   (d.) 17α-propyl estra-5 (10) -en-17β-ol-3-one (5):   Enol ether in THF (100 mL) and methanol (100 mL)4(3.00g, 9.10 Solution of oxalic acid (4.0 g) in H₂O solution (40 mL) was added. After 60 minutes at room temperature , A small amount of K₂CO_ThreeAnd add the resulting solid to a minimum of H 2.₂Dissolved in O. The mixture It was extracted with diethyl ether (3 x 100 mL). The combined ether layer is H₂O (50 mL) And washed with brine (2 x 50 mL), Na₂SO_FourThen dry and evaporate To give a white amorphous solid. In a subsequent step, this was diethyl ether / acetate Recrystallized from chill / hexane, white crystals (2.76g, 96%, melting point 148 ℃ -149.5 ℃) I got HRMS. C_{twenty one}H₃₄O₂Calculated value for: 316.240. Found: 316.240.   (e.) 17α-Propyl estra-4,9-dien-17β-ol-3-one (II-1):   γ-enone5(398 mg, 1.26 mmol) in pyridine (30 mL) was added with pyridine hydrate. Robromide perbromide (0.60 g, 1. 6 eq) was added. After 2 hours at room temperature, the reaction mixture was treated with 10% aqueous HCl (150 mL). Stir and extract with diethyl ether (3 x 25 mL). 10 combined ether layers % HCl aqueous solution (10 mL), brine (2 × 10 mL), saturated NaHCO 3._ThreeAqueous solution (2 x 25 mL) , Saturated CuSO_FourWash with aqueous solution (15 mL), and brine (2 x 15 mL), wash with Na₂SO_Fourso Dried and evaporated to give a yellow oil. Use this with silica gel Further purification was done by flash column chromatography. Ethyl acetate and And diene II-1 as a colorless foam (249 mg, 63%). Isolated. HRMS. C_{twenty one}H₃₀O₂Calculated value for: 314.225. Found: 314.225. UV (ethanol): λ_max306 nm (ε = 20,740).                               Example 2   In this example, 2 ', 3'α-tetrahydrofuran-2'-spiro-17- (5α-andross The preparation of tan [2,3-c] furazane (III-1) is described.   (a.) 2-hydroxyimino-17β-hydroxy-5α-androstan-3-one (7):   steroid6(17β-hydroxy-5α-androstan-3-one) benzene / me To the tanol solution was added a mixture of HCl and methanol. With vigorous stirring To this solution, t-BuNO in benzene₂Was added. During the addition, the reaction temperature was 19 ℃ and 2 Keep between 5 ° C. Also, t-BuNO₂Regarding the addition of, a precipitate appeared after the addition was completed. , And the reaction was stirred at room temperature for another hour. 20 mL H₂Add O and get Collected precipitates, H₂O, 2% NaHCO_ThreeAnd H₂Washed with O. After drying, 29.3 g of crude product7I got This was recrystallized from methanol and purified to 24.22 g. The product was obtained. The melting point was 264 ° C to 266 ° C (literature value; melting point 266 ° C to 267 ° C).   (B.) 17β-acetoxy-2,3-dihydroxyimino-5α-androstane (8):   steroid7, NH₂OH / HCl, and NaOAc / H₂The O 2 suspension was refluxed for 20 minutes. The reaction is cooled to room temperature and H₂It poured into O. The precipitate was collected by filtration. Get Solid8, H₂Washed with O and air and dried; weight 23.4g.   (C.) 17β-Hydroxy-5α-androstane [2,3-c] furazane (9):   Steroids in ethylene glycol8And KOH suspension at 180 ℃ -190 ℃ for 0.5 hours Heated for a while. During this time the steroid dissolved. The reaction mixture was cooled, then H 2₂O Poured into. The solid is collected by filtration and H₂Washed with O. Solid as CHCl_ThreeDissolved in Solved, Na₂SO_FourDried in vacuo and evaporated under reduced pressure. The weight is 7.49g Was. Dry column chromatography CHCl_ThreePurified by / 10% ethyl acetate, 3.3 0 g of pure product9I got Recrystallize from 95% ethanol to give 2.55g of product Obtained: melting point 157 ° C-159 ° C (literature value; melting point 160 ° C-161 ° C).   (D.) 17-keto-5α-androstane [2,3-c] furazan (Ten):   alcohol9(1.4 g, 4.43 mmol) in acetone solution (70 mL), Jones reagent Was added dropwise so that the orange color remained. The TLC at this time indicates the presence of starting material. I didn't. The reaction mixture was diluted with water (200 mL). Filter the crude material on the filter. Collected and washed with water. The crude material was dried overnight under reduced pressure and washed with silica gel. Chromatography on 5% ethyl acetate in dichloromethane, 10% hex Eluted with sun. pureTenYield, 1.19 g (85.5%).   (E.) 17β-hydroxy-17α-tetrahydropyranyl-oxypropargyl propargyl) -5α-Androstan [2,3-c] flazan (11):   THF in ethyl magnesium bromide (3.06 mL of 3.0 M in ether, 2.5 eq) Add the solution (10 mL) to the tetrahydropyra of propargyl alcohol at -78 ° C. Nyl ether (1.47 mL, about 2.6 eq) was added via syringe. The resulting slurry Warm to room temperature over 1 hour and use ketoneTen(1.19 g, 3.79 mmol) in THF (15 mL) was added using a cannula. Very little starting material remained after 10 minutes. Room After stirring at room temperature for 8 hours, the reaction mixture was mixed with saturated aqueous ammonium chloride solution (100 mL). Stir and CH₂Cl₂Extracted with (3 x 50 mL). Bored organic layer Wash with Japanese NaCl, K₂CO_Three2.47 g of yellow oyster dried and evaporated Got the le. It is purified on a silica gel column and CH₂Cl₂10% ethyl acetate in 1.72 desired product, eluting with / 10% hexane11Was obtained (yield about 100%).   (f.) 17β-Hydroxy-17α- (3′-tetrahydropyranyl-oxypropyl) -5α -Androstan [2,3-c] Frazan (12):   115% Pd / C (100 mg) in ethyl acetate solution (100 mL) of (1.72 g, 3.79 mmol) Was added and the solution was hydrogenated at atmospheric pressure. After 13 hours, starting material present No, some olefin remained. Add some new catalysts However, no further changes were observed after a further 2 hours. By filtering the catalyst Removed and the filter washed with ethyl acetate (100 mL). Evaporate the filtrate To obtain a white foam. 1.61 g of crude product12I got   (g.) 17β-hydroxy-17α- (3'hydroxypropyl) -5α-androstane [2, 3-c] Frazan (13):   12To a methanol solution (100 mL) of (crude product 1.6 g, 3.49 mmol) was added p-toluene. Sulfonic acid (0.72 g, 1.1 eq) was added. After stirring for 30 minutes at room temperature, TLC is allowed to react. Is complete. The reaction solution was saturated aqueous sodium bicarbonate solution (100 mL). And CH₂Cl₂Stirred with (100 mL). The resulting suspension was poured into water (50 mL). water CH solution₂Cl₂It was extracted with (2 × 100 ml). CH together₂Cl₂Saturated sodium chloride Solution (50 mL), dried over potassium carbonate, and evaporated to 1. 22 g of white solid was obtained. NMR is the intended structure13Matched.   (h.) 2 ', 3'α-tetrahydrofuran-2'-spiro-17- (5α-androstane [2,3-c ] Frazan (III-1):   Diol13To a pyridine solution (100 mL) of (1.22 g, 3.26 mmol) was added p-toluene. Sulfonyl chloride (2.0 g; 3.2 eq) was added under argon. 16 at room temperature Stir for hours to give the desired product III-1. Reaction mixture was saturated with aqueous sodium bicarbonate Washed with solution (150 mL) for 10 minutes. The suspension obtained is treated with water and dried. Extracted with ether (3 x 100 mL). The combined ether layer was washed with sodium bicarbonate. And washed with NaCl solution, dried over potassium carbonate and evaporated to give 0.98 g of crystals. Material was obtained. Purify III-1 by column chromatography, 0.39g white Plates (melting point 152 ° C-153 ° C) were obtained. The structure was analyzed by NMR, UV and IR spectroscopy. Confirmed.                                 Example 3                               Biological evaluation Preparation of cytosol:   Male rats were castrated 20-22 hours prior to sacrifice. Decapitate and exsanguine these animals Was slaughtered by. Immediately after the prostate was removed from the animal, it was kept on ice. Fat After removal, the prostate is minced into 3 volumes of Tris-HCl buffer (0.02M, pH 7. 4, containing 1.5 mM EDTA and 10% glycerol). Homozygous The nates were centrifuged at 10,000 xg for 10 minutes, and the resulting supernatant was 60 at 100,000 xg. Centrifuged for minutes. The final supernatant contained androgen binding receptor. Labeling of receptor proteins:   For the binding assay, 0.75 mL of prostate cytosol was added at 0.09 ng, 27500 dpm.^ThreeH -Only 5 μl DMSO containing dihydrotestosterone and 1 μL DMSO Was mixed with DMSO + competitor. Use this mixture to homogenize the tissue It was made up to 1.0 mL with Tris-HCl buffer. The mixture is then shaken occasionally at 4 ° C. for 4 hours. While incubating. At the end of the incubation, receptor binding Androgens were isolated by ammonium sulfate precipitation. Precipitation of receptor-bound androgen:   Prepare a 70% saturated ammonium sulfate solution and adjust its pH to 7.4 with aqueous ammonia. Was. Gently add 1.0 mL of 70% saturated ammonium sulfate to 1.0 mL of labeled cytosol. It was added dropwise with stirring. Allow the sample to sit at 0-4 ° C for 35 minutes, then at 10,000 xg Centrifuge for 20 minutes. The precipitate was treated with Tris-HCl as described above containing 2 mg bovine albumin. Resuspended in 1.0 mL of buffer. The androgen receptor complex is saturated with 70% of its equivalent volume. It was immediately reprecipitated by the addition of ammonium sulphate. Samples at 0-4 ° C It was left for 45 minutes and then centrifuged again as above. Decant the supernatant and The tube wall was completely dried with filter paper. 1.0 mL of final precipitated protein Resuspend in 10 mL of the same buffer and then add 10 mL of Scintisol for counting. Quantitatively transferred to a counting vial. The androgen receptor complex is 35% sulfate Precipitation with ammonium and release^ThreeNot only isolated from H-dihydrotestosterone And androgen binding protein precipitated with 50% ammonium sulfate, and 60-8 It is also isolated from bovine albumin precipitated with 0% ammonium sulfate. Comparison of relative binding affinities:   Comparing the relative binding affinities of different antianthrogens reported in the literature For this, we have used dihydrtestosterone (DHT) as a standard and Setting the value of to 100, the reported IC for various antiandrogens₅₀Values from those Relative binding affinity was estimated. This comparison is shown in Table 1. As the table shows, Products II-1 and III-1 are cyprotease receptors for the rat prostate androgen receptor. Exhibits a higher affinity than all other known antiandrogens except for long acetate .   In addition, compounds II-1 and III-1 are the rat ventral prostate androgen receptors It can be seen to show compatibility. Compounds II-1 and III-1 contain three other non-stero Flutamide (an anti-androgen receptor antagonist that acts as an anti-androgen receptor) Marketed in the United States), nilutamide (developed by Roussel, France) , Combine. Androgen receptors, as previously shown by other antiandrogens The extent of binding of the thyroid gland by inhibition of prostate growth in uncastrated male rats. Shows good correlation with in vivo antiandrogenic potency, as measured It is interesting to mean that.   Previous in castrated male rats given additional testosterone, summarized below. In an in vivo study evaluating compounds II-1 and III-1 for inhibition of gonad growth Is a similar correlation between in vivo potency and androgen receptor binding affinity The relationship is clear as shown in Table 3. Inhibition of Prostate Growth by Compounds II-1 and III-1 in Castrated Male Rats:   Male rats weighing 36-38 grams were castrated at 22 days of age. On the day of castration, agonize the animals Strtestosterone 0.6 mg; and antagonist III-1, 30.0 mg; II-1, 30.0 mg And 30.0 mg cyproterone acetate as the anti-androgen reference standard Used. Castrated rats were treated with steroids once daily for 10 consecutive days. place On day 10 post-procedure, the rats were sacrificed and dissected, and the final seminal vesicle weight and ventral anterior The gland weight was measured. Compounds II-1 and III-1 and anti-androgen reference standard The results of inhibition of prostate growth by cyproterone acetate are shown in Table 2 below.   These results clearly indicate that compounds II-1 and III-1 are steroidal anti-and Established as a new chemical class of Rogen. In vivo, they are Acts as a drogen receptor antagonist and in castrated male rats And inhibits prostate growth stimulated by exogenous testosterone. Prostate growth The degree of inhibition also appeared to be dose-dependent, as shown in Table 4, but this The trial conclusions of this study were that further dose / response experimental studies were not conducted for final confirmation. I have to.   The relative anti-androgen potency in vivo is as follows:   Different Uses in the Paragonad and Testis of Uncastrated Sprague-Dawley Male Rats Effects of Amount of Antiandrogen Compound III-1:   Using the above procedure, the non-castrated Sprague-Dawley male rat paragonads and The effect of anti-androgen compound III-1 on the testis was evaluated. The results are shown in Table 4. . As can be seen in the table, anti-androgens of the Ventral Prostate Growth After Administration Antagonism on the Effect of Testosterone Produced Is shown.   Agonist Andro of Competitive Receptor Binding Antagonists II-1 and III-1 Lack of gen activity was performed on 22-day-old castrated male rats using standard Hershberger and It was shown in vivo by a rogen assay. I at doses of 0.8 mg, 1.6 mg, and 3.2 mg The weight of seminal vesicles and ventral prostates of animals treated with I-1 and III-1 is vehicle only Was equal to the control of. Very flat response to androgenic activity The line was obtained. It is estimated that this is an effective anti-inflammatory agent for the treatment of castrated male rats. At the androgen dose (30.0 mg) both III-1 and II-1 are essentially agonist agonists. Shows a lack of drogenic activity.   FIG. 1 shows that Compounds II-1 and III-1 produced fairly stable prostate weight at higher doses. It clearly shows that even in the case of. By comparison, testosterone itself It can be seen that the body proportionally increases the weight of the prostate with increasing dose . Prostate C_ThreeMeasurement of mRNA:   The Prostate 17: 41 (1990), dihydrotestosterone Of rat ventral prostate explant culture for compound III-1 combined with Drogen responsiveness was evaluated. Testosterone was used as a control. Said In the trial, as described in the reference, prostatin (rat ventral prostate More secreted androgen-dependent protein). Rat prostate ec Addition of androgen to the culture medium of sprung culture was carried out by adding C of prostate._Threesub Maintains the unit's mRNA, and prostatein or its C_ThreeSubunit mRN Any measurement of A is an androgen response sensitive to the explant system. Useful as an answer marker. The results are shown in Table 5: As can be seen in Table 5, prostatin and subunit mRNAs are post-castration , Which is significant in the compound III-1 / dihydrotestosterone formulation. It exhibits anti-androgenic activity. Additional in vivo studies:   An additional in vivo protocol that can assess the efficacy of novel compounds is the male Copenhagen. × Fischer's F₁Androgen-sensitive PAP mutation in rat Dunning R-3327 prostate tumor Includes the body. Use flutamide, nilutamide, etc. as positive controls As a novel compound, administered by oral gavage and subcutaneous sesame oil suspension Can be done. Once the tumor is measurable (generally 7-20 weeks after transplantation), tumorigenicity Can be tested. 20, 60, and 200 mg (subcutaneous) / kg / day in 3 treatment groups Feed, 4th group 100 mg (oral) / kg / day, and 5th control 30 mg (subcutaneous) / kg / day of flutamide or the same dose of nilutami for the roll group Give the de. Rats receive a single daily dose either subcutaneously or by oral gavage Receive 28 days and weigh weekly. Tumors measured 3 times twice a week Set. Tumor volume doubling time and ventral prostate weight are evaluated. Compound of the present invention Significantly reduces tumor volume doubling time and keeps ventral prostate weight relatively low It is expected that.

────────────────────────────────────────────────── ─── Continuation of front page    (72) Inventor Chung, Wesley Kay. M.             United States California 92024,               Encinitas, W. "Eye"               Street number 3 330 (72) Inventor Clau, David F.             United States California 93726,               Fresno, East Andrews             2615 E

Claims (1)

[Claims] 1. A compound having the following structural formula (II): Wherein R ¹ and R ² are independently selected from the group consisting of hydrogen and lower alkyl; one of R ⁶ and R ⁷ is hydroxyl and the other is n-propyl, or R ⁶ and R 7 ⁷ is joined to provide a cycloalkyl or cyclooxyalkyl group containing 3 to 6 carbon atoms; and R ⁸ and R ⁹ are hydrogen. 2. The compound of claim 1, wherein one of R ⁶ and R ⁷ is hydroxyl and the other is n-propyl. 3. The compound of claim 2, wherein R ⁶ and R ⁷ are joined to provide a cycloalkyl or cyclooxyalkyl group containing 3 to 6 carbon atoms. 4. A compound according to claim 2 having the following structural formula (II-1): 5. A compound having the following structural formula (III): Wherein R ⁵ is hydrogen or lower alkyl; one of R ⁶ and R ⁷ is hydroxyl and the other is n-propyl, or R ⁶ and R ⁷ are joined to form 3 to 6 Providing a cycloalkyl or cyclooxyalkyl group containing carbon atoms; and R ⁸ and R ⁹ are hydrogen. 6. The compound of claim 5, wherein one of R ⁶ and R ⁷ is hydroxyl and the other is n-propyl. 7. The compound of claim 5, wherein R ⁶ and R ⁷ are joined to provide a cycloalkyl or cyclooxyalkyl group containing 3 to 6 carbon atoms. 8. The compound of claim 6, having the following structural formula (III-1): 9. A pharmaceutical composition comprising an amount of a compound having the structural formula (II) below in combination with a pharmaceutically acceptable carrier: Wherein R ¹ and R ² are independently selected from the group consisting of hydrogen and lower alkyl; one of R ⁶ and R ⁷ is hydroxyl and the other is n-propyl, or R ⁶ and R 7 ⁷ is joined to provide a cycloalkyl or cyclooxyalkyl group containing 3 to 6 carbon atoms; and R ⁸ and R ⁹ are hydrogen. 10. The composition according to claim 9, wherein in the compound (II), one of R ⁶ and R ⁷ is hydroxyl and the other is n-propyl. 11. The composition according to claim 10, wherein in the compound (II), R ⁶ and R ⁷ are combined to provide a cycloalkyl group or a cyclooxyalkyl group containing 3 to 6 carbon atoms. 12. The composition of claim 10, wherein the compound has the following structural formula (II-1): 13. A pharmaceutical composition containing an amount of a compound having structural formula (I II) below in combination with a pharmaceutically acceptable carrier: Wherein R ⁵ is hydrogen or lower alkyl; one of R ⁶ and R ⁷ is hydroxyl and the other is n-propyl, or R ⁶ and R ⁷ are joined to form 3 to 6 Providing a cycloalkyl or cyclooxyalkyl group containing carbon atoms; and R ⁸ and R ⁹ are hydrogen. 14． The composition according to claim 13, wherein in the compound (III), one of R ⁶ and R ⁷ is hydroxyl and the other is n-propyl. 15. The composition according to claim 14, wherein in the compound (III), R ⁶ and R ⁷ are combined to provide a cycloalkyl group or a cyclooxyalkyl group containing 3 to 6 carbon atoms. 16. 15. The composition of claim 14, wherein the compound has the structural formula (III-1):