MXPA98009292A - Benzotiophenes, formulations that contain them, ymeto - Google Patents

Abstract

The invention provides the compounds of the formula I, wherein R 1 is hydrogen, hydroxyl, alkoxy of 1 to 4 carbon atoms, -OCOO (alkyl of 1 to 6 carbon atoms), -OCO (alkyl of 1 to 6 carbon atoms), -OCOAr wherein Ar is phenyl or optionally substituted phenyl, -OSO2 (straight chain alkyl of 4 to 6 carbon atoms), or -OSO3H, R2 is R1, Cl or F, with the proviso that at least one of R1 or R2 is -OSO3H; R3 is 1-piperidinyl, 1-pyrrolidinyl, methyl-1-pyrrolidinyl, dimethyl-1-pyrrolidino, 4-morpholino, dimethylamino, diethylamino, diisopropylamino, or 1-hexamethyleneimino; and n is 2 or 3; pharmaceutically acceptable solvates thereof. The present invention further relates to pharmaceutical compositions containing the compounds of the formula (I), optionally containing estrogen or progestin, and the use of such compounds, alone, or in combination with estrogen or progestin, to alleviate the symptoms of post-menopausal syndrome, particularly osteoporosis, pathological conditions related to the cardiovascular system, and estrogen-dependent cancer. The compounds of the present invention are also useful for inhibiting uterine fibroid disease and endometriosis in women and the proliferation of aortal smooth muscle cells, particularly restenosis, in human

Description

BENZOTIOPHENES, FORMULATIONS THAT CONTAIN THEM, AND METHODS FIELD OF THE INVENTION This invention relates to the fields of pharmaceutical and organic chemistry - and provides benzothiophene compounds, which are useful for the treatment of various medical indications associated with post-menopausal syndrome, and uterine fibroid disease, endometriosis, and proliferation of smooth aortic muscle cells. The invention also relates to pharmaceutical compositions.

BACKGROUND OF THE INVENTION "Post-menopausal syndrome" is a term used to describe various pathological conditions that frequently affect women who have entered or completed the physiological metamorphosis known as menopause. Although numerous pathologies are contemplated by the use of this term, three major effects of the post-menopausal syndrome are the source of the greatest medical interest to REF: 28605 long-term: osteoporosis, cardiovascular effects such as hyperlipidemia, and cancer-dependent estrogen, particularly breast and uterine cancer. Osteoporosis describes a group of diseases that arise from various etiologies, but which are characterized by the net loss of bone mass per unit volume. The consequence of this loss of bone mass and resulting bone fracture is the failure of the skeleton to provide adequate structural support for the body. One of the most common types of osteoporosis is that associated with menopause. Most women lose from about 20% to about 605 of the bone mass in the trabecular compartment of the bone within 3 to 6 years after cessation of menses. This rapid loss is generally associated with an increase in bone resorption and formation. However, the restoring cycle is more dominant and the result is a net loss of bone mass. Osteoporosis is a common and serious disease among post-menopausal women. There is an estimate of approximately 25 million women in the United States, only, who are affected with this disease. The results of osteoporosis are personally harmful and also explain a great economic loss due to its chronicity and the need for long-term and extensive support (hospitalization and nursing home care) due to the consequences of the disease. This is especially true in older patients. In addition, although it is not generally thought that osteoporosis is a life-threatening condition, a mortality rate of 20% to 30% is related to hip fractures in elderly women. A large percentage of this mortality rate may be directly associated with post-menopausal osteoporosis. The most vulnerable tissue in the bone for the effects of postmenopausal osteoporosis is trabecular bone. This tissue is often referred to as cancellous or cancerous bone and is particularly concentrated near the ends of the bone (near the joints) and in the vertebrae of the spine. The trabecular tissue is characterized by small osteoid structures which are interconnected with each other, as well as the more solid and dense cortical tissue which constitutes the outer surface and the central axis of the bone. This interconnected network of trabeculae gives lateral support to the external cortical structure and is critical for the biomechanical strength of the entire structure. In post-encephalic osteoporosis, it is mainly net resorption and loss of trabeculae that lead to bone failure and fracture. In light of the loss of the trabeculae of post-menopausal women, it is not surprising that most fractures are those associated with bones that are highly dependent on the trabecular support, for example the vertebrae, the neck of the bones that they support weight, such as the femur and the forearm. Of course, hip fracture, Colles fractures, and vertebral crush fractures are hallmarks of post-menopausal osteoporosis. At this time, the most predominant method for the treatment of postmenopausal osteoporosis is estrogen replacement therapy. Although this therapy is generally successful, the patient's compliance with the therapy is mainly low because the treatment with estrogen frequently produces undesirable side effects. Recently, a new treatment for post-menopausal osteoporosis has been achieved, for example, treatment with bis-phosphonates. Although this therapy is effective, it suffers from the disadvantage of treating only the aspect of osteoporosis of post-menopausal syndrome. In addition, many reports on the use of bisphosphonates have revealed undesirable gastrointestinal side effects. Throughout post-menopausal time, most women have a lower incidence of cardiovascular disease than men of the same age. After menopause, however, the rate of cardiovascular disease in women increases slowly to equal the rate observed in men. This loss of protection has been linked to the loss of estrogen and, in particular, to the loss of estrogen's ability to regulate serum lipid levels. The nature of estrogen's ability to regulate serum lipids is not yet well understood, but evidence to date indicates that estrogen can up-regulate low-density lipid (LDL) receptors in the liver to eliminate excess of cholesterol. In addition, estrogen seems to have some effect on the biosynthesis of cholesterol, and other beneficial effects on cardiovascular health. It has been reported in the literature that post-menopausal women who have estrogen replacement therapies have a return of serum lipid levels to concentrations of the pre-menopausal state. In this way, estrogen would seem to be a reasonable treatment for this condition. However, the side effects of estrogen replacement therapy are not acceptable to many women, thus limiting the use of this therapy. An ideal therapy for this condition would be an agent that could regulate the level of serum lipids as estrogen does, but that would be devoid of the side effects and risks associated with estrogen therapy. The third major pathology associated with post-menopausal syndrome is estrogen-dependent breast cancer and, to a lesser extent, estrogen-dependent cancers of other organs, particularly the uterus. Although such neoplasms are not only limited to a postmenopausal woman, and the use of the compounds herein is not limited to such, they are more prevalent in the older post-menopausal population. Current chemotherapy of these cancers has relied heavily on the use of anthogenic compounds such as, for example, tamoxifen. Although such mixed agonist-antagonists have beneficial effects in the treatment of these cancers, and the estrogenic side effects are tolerable in acute life-threatening situations, these are not ideal. For example, these agents may have stimulatory effects on certain populations of cancer cells in the uterus, due to their estrogenic properties (agonists) and may, therefore, be counterproductive in some cases. A better therapy for the treatment of these cancers would be an agent that is an antiestrogenic compound that has negligible or non-estrogenic agonist properties on the reproductive tissues. In response to the clear need for new pharmaceutical agents that are capable of alleviating the symptoms of, among other things, post-menopausal syndrome, the present invention provides benzothiophene compounds, pharmaceutical compositions thereof, and methods for using such compounds for the treatment of post-menopausal syndrome and other pathological conditions related to estrogen, such as those mentioned below. Uterine fibrosis (uterine fibroid disease) is an old and even current clinical problem that presents a variety of names, including uterine fibroid disease, uterine hypertrophy, uterine lieomyomata, myometrial hypertrophy, fibrous uterus, and fibrotic metritis. Essentially, uterine fibrosis is a condition where there is an inappropriate deposition of fibroid tissue on the wall of the uterus. This condition is a cause of dysmenorrhea and infertility in women. The exact cause of this condition is poorly understood, but the evidence suggests that it is an inappropriate response of fibroid tissue to estrogen. Such a condition has been produced in rabbits by the daily administration of estrogen for 3 months. In guinea pigs, the condition has been produced by the daily administration of estrogen for 4 months. In addition, in rats, estrogen causes similar hypertrophy. The most common treatment of uterine fibrosis involves surgical procedures that are either expensive and sometimes a source of complications such as the formation of adhesions and abdominal infections. In some patients, the initial surgery is only a temporary treatment and the fibroids develop again. In those cases a hysterectomy is performed which effectively ends with the fibroids but also with the patient's reproductive life. Also, gonadotropin releasing hormone antagonists can be administered, but their use is diminished by the fact that they lead to osteoporosis. Thus, there is still a need for new methods for the treatment of uterine fibrosis, and the methods of the present invention satisfy that need. Endometriosis is a condition of severe smenorrhea, which is accompanied by severe pain, bleeding within the endometrial masses or peritoneal cavity and often leads to infertility.The cause of the symptoms of this condition seems to be endometrial development. Ectopics that respond inappropriately to normal hormonal control and are located in inappropriate tissues.Due to inappropriate locations for endometrial development, the tissue seems to initiate local responses similar to inflammatory ones causing macrophage infiltration and a cascade of events leading to the onset of The painful response The exact etiology of this disease is not well understood and its treatment by hormonal therapy is diverse, poorly defined, and marked by numerous unwanted and perhaps dangerous side effects One of the treatments for this disease is the use of estrogen at low doses to suppress the endometrial development through a negative feedback effect on the release of central gonadotropin and the subsequent production of estrogen in the ovary; however, it is sometimes necessary to use continuous estrogen to control the symptoms. This use of estrogen can often lead to undesirable side effects and even the risk of endometrial cancer. Another treatment consists of the continuous administration of progestins which induce amenorrhea, and by suppressing the production of ovarian estrogens they can cause regressions of the endometrial developments. The use of chronic progestin therapy is often accompanied by the side effects of progestins, which are unpleasant in the central nervous system, and often leads to infertility due to the suppression of ovarian function. A third treatment consists of the administration of weak androgens, which are effective in controlling endometriosis; however, they induce severe effects of masculinization. Several of these treatments for endometriosis have also been implicated in causing a slight degree of bone loss with continuous therapy. Therefore, new methods of treating endometriosis are desirable. The proliferation of smooth aortic muscle cells plays an important role in diseases such as atherosclerosis and restenosis. Vascular restenosis after percutaneous transluminal coronary angioplasty (PTCA) has been shown to be a tissue response characterized by an early phase and a late phase. The early phase that occurs hours to days after PTCA is due to thrombosis with some vasospasms, whereas the late phase appears to be dominated by excessive proliferation and migration of aortal smooth muscle cells. In this disease, increased cell mortality and colonization by such muscle cells and macrophages contribute significantly to the pathogenesis of the disease. Excessive proliferation and migration of vascular aortic smooth muscle cells may be the primary mechanism for reocclusion of the coronary arteries after PTCA, atherectomy, laser angioplasty and arterial bypass graft surgery. See "Intimal Proliferation of Smooth Muscle Cells as an Explanation for Recurrent Coronary Artery Stenosis after Percutaneous Transluminal Coronary Angioplasty," Austin et al., Journal of the American College of Cardiology, 8: 369-375 (August 1985). Vascular restenosis remains a major long-term complication after surgical intervention of blocked arteries, through percutaneous transluminal coronary angioplasty (PTCA), atherectomy, laser angioplasty, and arterial bypass graft surgery. In approximately 35% of patients who suffer from PTCA, reocclusion occurs within three to six months after the procedure. Current strategies for treating vascular restenosis include mechanical intervention by devices such as stents or drug therapies including heparin, low molecular weight heparin, coumarin, aspirin, fish ceite, calcium antagonist, steroids and prostacyclin. These strategies have failed to decrease the reocclusion rate and have not been effective for the treatment and prevention of vascular restenosis. See "Prevention of Restenosis after Percutaneous Transluminal Coronary Angioplasty: The Search for a Magic Bullet," Hermans et al., Ameri can Heart Journal, 122: 171-187 (July 1991). In the pathogenesis of restenosis, proliferation and excessive cell migration occur as a result of the growth factors produced by the cellular constituents in the blood and the damaged wall of the arterial vessels, which mediates the proliferation of the smooth muscle cell in vascular restenosis. Agents that inhibit the proliferation and / or migration of aortal smooth muscle cells are useful in the treatment and prevention of restenosis. The present invention provides the use of compounds as inhibitors of the proliferation of the aortic muscle cell and, thus, inhibitors of restenosis.

BRIEF DESCRIPTION OF THE INVENTION This invention provides the compounds of the formula I (I. wherein R1 is hydrogen, hydroxyl, alkoxy of 1 to 4 carbon atoms, -OCOO (alkyl of 1 to 6 carbon atoms), -OCO (alkyl of 1 to 6 carbon atoms), -OCOAr where Ar is phenyl or optionally substituted phenyl, -OS02 (straight chain alkyl of 4 to 6 carbon atoms), or -0S03H; R2 is R1, Cl or F; with the proviso that at least one of R1 or R2 is -OSO3H; R3 is 1-piperidinyl, 1-pyrrolidinyl, methyl-1-pyrrolidinyl, dimethyl-1-pyrrolidino, 4-morpholino, dimethylamino, diethylamino, diisopropylamino, or 1-hexamethyleneimino; and n is 2 or 3; or a pharmaceutically acceptable salt or solvate thereof. The present invention further relates to pharmaceutical compositions containing the compounds of formula I, which optionally contain estrogen or progestin, and the use of such compounds, alone, or in combination with estrogen or progestin, to alleviate the symptoms of post-stroke syndrome. -menopausal, particularly osteoporosis, pathological conditions related to the cardiovascular system, and estrogen-dependent cancer. As used herein, the term "estrogen" includes steroid compounds that have estrogenic activities, such as, for example, 17-b-estradiol, estrone, conjugated estrogen (Premarin®), equine estrogen, 17-b-ethinylestradiol , and similar. As used herein, the term "progestin" includes compounds that have progestational activity such as, for example, progesterone, norethylnodrel, nongestrel, megestrol acetate, norethindrone, and the like. The compounds of the present invention are also useful for the inhibition of uterine fibroid disease and endometriosis in women and the proliferation of aortal smooth muscle cell, particularly restenosis, in humans.

DETAILED DESCRIPTION OF THE INVENTION The present invention is related to the discovery that a select group of 2-aryl-3-arylbenzo [b] thiophenes, for example, the compounds of the formula I, are useful for the treatment or prevention of the symptoms and pathologies of: post-menopausal-osteoporosis and hyperlipidemia, estrogen-dependent cancers, uterine fibrosis, endometriosis, or restenosis in mammals, including humans The term inhibit is defined to include its generally accepted meaning which includes prohibition, prevention, restriction, and retardation, arrest or progression of the reversion, or severity, or such action on a resulting symptom As such, the present invention includes medical therapeutic and / or prophylactic administration, as appropriate In the above formula, the term "alkyl" of 1 to 6 carbon atoms "represents a linear or branched alkyl chain having from 1 to 6 carbon atoms. 1 to 6 carbon atoms include methyl, ethyl, n-propyl and n-butyl. The term "alkoxy of 1 to 4 carbon atoms" represents groups such as methoxy, ethoxy, n-propoxy, and n-butoxy. Optionally substituted phenyl includes phenyl and phenyl substituted once or twice with alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 4 carbon atoms, hydroxyl, nitro, chloro, fluoro, or tri (chloro or fluoro) methyl. The term "solvate" represents an aggregate comprising one or more molecules of the solute, such as a compound of the formula I, with one or more molecules of the solvent. The starting material for the preparation of the compounds of the present invention is a compound of the formula II II where Rla is hydrogen, hydroxyl, or alkoxy of 1 to 4 carbon atoms, R2a is hydrogen, hydroxyl, -Cl, -F, or alkoxy of 1 to 4 carbon atoms, with the proviso that at least one of Rla or R2a is hydroxyl; and R3 and n have their previous meanings. The compounds of formula II are well known in the art and are prepared essentially as described in U.S. Patent Nos. 4,133,814, 4,418,068, and 4,380,635 all of which are incorporated by reference herein. See, also, Jones, C.D. and collaborators, J. Med. Chem. , 27: 1057-66 (1984).

In the preparation of the compounds of the present invention, in general, a monophenolic compound of the formula II is sulfated to provide an acid sulfate derivative of the formula la. The monosulfate product can be purified as such, thereby producing the neutral amphoteric form. This can also be converted to a variety of pharmaceutically acceptable salts. the wherein Rlb is hydrogen, alkoxy of 1 to 4 carbon atoms, or -0S03H; R2b is hydrogen, alkoxy of 1 to 4 carbon atoms, Cl, -F or -OS03H, with the proviso that R Ib or R2 must be -OS03H; and R "and n have their previous meanings Other compounds (formula Ib) of the present invention can be prepared from the compounds of formula II, where Rla and R2a are both hydroxyl These compounds can be monosulphated using a simple equivalent of sulfation reagent and a simple equivalent of a strong base to ionize one of the phenols that will produce a mixture of isomers The resulting mixture of derivatives (mono-sulphate-ono-hydroxy) can be directly isolated by techniques such as precipitation or recrystallization , which are well known to those of skill in the art Alternatively, the derivatives can be purified by normal phase or reverse phase chromatography.The mono-sulfate mono-hydroxy derivatives, thus obtained, can also be converted to other compounds of the formula Ib by appropriate acylation or sulfonation of the phenolic hydroxyl by the methods two known in the art. Such methods may be found in the references cited above or in U.S. Patent No. 5,482,949. By means of such chemical transformations, the compounds of the formula la and Ib, which together constitute the compounds of the formula I, can be obtained.

Ib wherein Ric is hydroxyl, -OCOO (alkyl of 1 to 6 carbon atoms), -OCO (alkyl of 1 to 6 carbon atoms), -OCOAr wherein Ar is phenyl or optionally substituted phenyl, -OS02 (chain alkyl) linear of 4 to 6 carbon atoms), or -OS03H; R2c is Rlc, with the proviso that at least one of Rlc or R2c is -0S03H; R3 is 1-piperidinyl, 1-pyrrolidinyl, methyl-1-pyrrolidinyl, dimethyl-1-pyrrolidino, 4-morpholino, dimethylamino, diethylamino, diisopropylamino, or 1-hexamethyleneimino; and n is 2 or 3. In the sulfation step of the present process, a phenolic compound of formula II is converted to its mono- (or di-) sulfate derivative, via a one-step protocol, essentially as described by Gilbert, EE et al., Ch emi cal Rev., 62: 549-589 (1962). In essence, a phenolic starting material of formula II is contacted with a sulfation reagent such as sulfur trioxide (S03), S03-pyridine, S03-trimethylamine, S03-triethylamine, S? 3-dimethylaniline, S03-dioxane , S03-thioxane, S03-2-methylpyridine, S03-quinoline, S03-dimethylformamide, S03-trimethylamine which is particularly preferred, and the like, in a suitable inert solvent, in the presence of an acid scavenger, such as a metal base alkaline or a tertiary amine. Suitable inert solvents include, for example, alcohols, ethers, polar solvents such as dimethylformamide or dimethyl sulfoxide and particularly water. A preferred alkaline solution for the sulfation reaction contains sodium or potassium hydroxide in an inert solvent such as water. In this solution, the hydroxyl group (s) of the initial phenol material of the formula II exist as a phenoxide ion which readily participates in the sulfation reaction by reaction with sulfur trioxide or a derivative thereof. When running under the preferred reaction conditions, the present sulfation reaction takes from about 12 to about 72 hours to complete. The following examples are presented to further illustrate the preparation of the compounds of the present invention. The invention is not intended to be limited in scope by virtue of any of the following examples. The nuclear magnetic resonance (NMR) data for the following examples were generated on a 300 MHz GE NMR instrument, and anhydrous d-6 DMSO was used as the solvent, unless otherwise indicated.

Example 1A [2- (4-hydroxy-enyl) -6-hydrogensulf-oyloxy-benzo [b] thien-3-yl] [4- [2- (1-piperidinyl) ethoxy] -fenyl] -methanone The hydrochloride salt of [6-hydroxy-2- (4-hydroxyphenyl) benzo [b] thien-3-yl] [4- [2- (l-piperidinyl) ethoxy-phenyl-methanone, (Raloxifene hydrochloride; 2.02 g, mmol) was mixed with 16 ml of IN sodium hydroxide, to provide a dark reddish-brown solution to which S 3 3 Me 3 N (0.566 g, 4 mmol) was added at room temperature under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 4 days, at which time thin layer chromatography (silica gel, 8: 1: 1 v / v chloroform methanol: triethylamine) indicated a mixture of starting material (Rf 0.7 ), the 4'-monosulfate (Example IB) (Rf 0.5), the 6-monosulfate (Example IA) (Rf 0.4), and the 4 ', 6-disulphate (Example 2) that had been formed. Chromatographic analysis by high performance liquid chromatography (HPLC) (C-18 Novapak column), 3.9 mm x 150 mm; with UV detection equipment at 280 nm, and a flow rate of 1 ml / minute of isocratic aqueous buffer of acetonitrile: 0.5% NH4H2P04, 20:80) indicated that the compounds were formed in a percentage ratio of area of 1.0 : 0.5: 1.2: 0.2, respectively. One quarter of the reaction mixture, which was a dark brown-yellow solution, was applied directly to two RPC18 Waters cartridges for purification on the LC2000 instrument. The cartridges were pre-equilibrated with aqueous buffer of acetonitrile: 0.5% NH4H2P04, 10:90 and 5 ml of deionized water was applied just before and just after the introduction of the sample to minimize the precipitation of the sample. The column was then eluted at a rate of 150 ml per minute with a gradient system which initially consisted of acetonitrile: 10:90 ammonium diacid phosphate buffer (as described above) and linearly raising to 40:60 acetonitrile: shock absorber . The elution was then continuously continued to a 40:60 mixture for an additional 10 minutes. Fractions of approximately 200 ml were collected and analyzed by high performance liquid chromatography, and appropriate fractions were combined. The rest of the reaction mixture was purified in batches in a similar manner. Although there was considerable overlap of the peaks corresponding to the two monosulfates, the eluent enriched in the 6-monosulfate was separated from that enriched in the 4'-monosulfate. The enriched fractions of each isomer were separately concentrated to near dryness and the residue was washed with water to remove the inorganic salts, and the sparingly soluble monosulfates were collected by filtration. In the final purification, each monosulfate was redissolved in aqueous NaOH (pH 11) and re-chromatographed essentially as already described. After the combination and concentration of the appropriate fractions, the precipitation procedure was repeated to provide 142 mg (7%) of [2- (4-hydroxyphenyl) -6-hydrogensulfoyloxybenzo [b] thien-3-yl] [4- [2- (1-piperidinyl) ethoxy] phenylmetanone as a whitish amorphous solid, with pink dyes which had a purity > 97% determined by the test of HPLC. : H NMR (DMSO-d6) d 9.79 (s, ÍH), 9.17 (s broad, ÍH), 7. 84 (d, J = 2.2 Hz, 1H), 7.73 (d, J = 8.7 Hz, 2H), 7. 30 (d, J = 8.7 Hz, ÍH), 7.24 (d, J = 8.7 Hz, 2H), 7. 19 (dd, J = 2.2 Hz, J = 8.7 Hz, ÍH), 6.99 (d, J, = 8. 7 Hz, 2H), 6.70 (d, J = 8.7 Hz, 2H), 4.38 (t, J = 4. 8 Hz, 2H), 3.48-3.33 (m, 4H), 2.97 (m, 2H), 1.80 (m, 2H), 1.65 (m, 3H), 1.37 (m, ÍH); MS (FAB "+ ion mode) m / e 554 (MH +); Calculated Analysis for C28H-27N0 S2: C, 60.74; H, 4.92; N, 2.53, Found C, 60.03; H, 5.02; N, 2.14.

Example IB [2 - (-4-Hydrogensul f or iloxy f eni l) - 6-hydroxybenzo [b] thien-3 i] [4- [2- (1-piperidinyl) ethoxyfine] methanone From Example IA, in a similar final purification procedure, rechromatography, concentration and precipitation, 196 mg (9%) of the title compound was obtained as a creamy amorphous solid with a purity >98% by HPLC analysis. H NMR (DMSO-d6) d 9.84 (s, 1H), 9.16 (broad s, 1H), 7.70 (d, J = 8.7 Hz, 2H), 7.38 (d, J = 2.2 Hz, 1H), 7.34 (d , J = 8.7 Hz, ÍH), 7.25 (d, J = 8.7 Hz, 2H), 7.08 (d, J = 8.7 Hz, 2H), 6.96 (d, J = 8.7 Hz, 2H), 6.90 (dd, J = 2.2 Hz, J = 8.7 Hz, ÍH), 4.35 (t, J = 4.8 Hz, 2H), 3.48 (m, 4H), 2.98 (, 2H), 1.80 (m, 5H), 1.66 (m, ÍH) 1.38 (m, ÍH); ' MS (FAB + ion mode) m / e 554 (MH +); Analysis Calculated for C i.H -N07S2: C, 60.74; H, 4.92; N, 2.53. Found C, 60.78; H, 5.11; N, 2.33.

Example 2 Triethylamine salt of [2- (4-hydrogensulfoyloxyphenyl) -6-hydrogensul-phenyloxybenzo [b] thien-3-yl] [4- [2- (l-piperidinyl) e t ox i] f enyl] met anone The hydrochloride salt of [6-hydroxy-2- (4-hydroxyphenyl) benzo [b] thien-3-yl] [4- [2- (l-piperidinyl) ethoxy-phenyl-methanone, (raloxifene hydrochloride; 1.02 g, 2.0 mmol) was combined with 8.0 ml of 1 N NaOH, and water (30 ml) and the mixture was heated under a nitrogen atmosphere in an oil bath at 60 ° C until a dark yellowish-brown solution was obtained. Without cooling the solution, S03-Me3N (1.11 g, 8.0 mmol) was added and heating was continued in a 60 ° C bath for 72 hours, during which time most of the dark color disappeared. HPLC assay using acetonitrile: 0.5% aqueous NH4H2P04 buffer 30:70 indicated a formation of more than 96% of the desired disulfate derivative. Less than 2% of the starting material or the monosulfates remained at the end of the reaction. The pale yellow reaction mixture was adjusted to pH 8.4 with 3N hydrochloric acid, filtered and applied directly to two cartridges of RPC18 (Waters LC2000 instrument, the cartridges had been pre-equilibrated with acetonitrile: 0.5% aqueous buffer of NH4H2P04 10: 90. Elution at a flow rate of 125 ml / min employed a gradient system that consisted initially of acetonitrile: aqueous buffer at 0.5% NH4H2P04 10:90, and then linearly changing the acetonitrile mixture in 40 minutes: buffer 30:70 and finally acetonitrile: buffer 50:50 in 5 minutes Fractions of approximately 200 ml each were collected and those considered to contain only 4 ', 6-disulfate were combined and evaluated by HPLC, which indicated purity greater than 98% The combined fractions were evaporated under reduced pressure and the temperature was kept below 30 ° C to eliminate most of the The acetonitrile The concentrate, which represented approximately 450 ml, was immediately applied to two Waters C18 cartridges and which had been pre-equilibrated with acetonitrile: water 10:90. Prolonged elution of the column with 3 liters of acetonitrile: water 10:90 was used to remove the inorganic salts. Later a linear gradient system that consisted initially of acetonitrile: water 10:90, and then changed linearly in 40 minutes to a mixture of acetonitrile: water 30:70 and finally to a 50:50 mixture of the same solvents in 5 minutes, it served to elute the desired product. The appropriate fractions (approximately 200 ml each) were combined to provide a total of approximately 1.1 liter which contained a single component by HPLC assay. A 300 ml aliquot of the combined fractions was treated at room temperature with a solution of 1 ml of triethylamine and 9 ml of water. The resulting clear and colorless solution was evaporated almost to dryness on a rotary evaporator while maintaining the temperature below 45 ° C. The colorless residue was dried under a high vacuum at room temperature to give an amorphous solid, 295 mg (74% yield), which was a simple component by HPLC analysis. lE NMR (DMSO-d6) d 9.16 (s broad, 1H), 8.82 (s broad H), 7.86 (d, J = 2.2 Hz, IH), 7.72 (d, J = 8.7 Hz, 2H), 7.38 (d , J = 8.7 Hz, ÍH), 7.31 (d, J = 8.7 Hz, 2H), 7.22 (dd, J = 2.2 Hz, J = 8.7 Hz, 1H), 7.10 (d, J = 8.7 Hz, 2H), 6.98 (d, J = 8.7 Hz, 2H), 4.37 (m, 2H), 3.55-3.32 (, 4H), 3.18-3.05 (m, 6H), 3.05-2.88 (m, 2H), 1.85-1.57 (m , 5H), 1.50-1.25 (m, ÍH); MS (FAB + ion mode) m / e 735.3 (MH +); An exact elemental analysis for carbon was not obtained: Analysis Calculated for C34H42 2O10S3: C, 55.57; H, 5.76; N, 3.81. Found C, 53.89; H, 5.67; N, 3.85.

The following examples illustrate the methods for the use of the compounds of the formula I in experimental models or in clinical studies.

Test Procedure General Preparation Procedure In the examples that illustrate the methods, a post-menopausal model was used in which the effects of the different treatments on the circulating lipids were determined. Female rats Sprague Da law of seventy-five days of age (weight range 200 to 225 g) were obtained from Charles River Laboratories (Portage, MI). The animals were bilaterally ovariectomized (OVX) or exposed to a fake surgical procedure at Charles River Laboratories, and then shipped after a week. After arrival, they were placed in hanging metal cages in groups of 3 or 4 per cage and had access to food ad l ibi t um (calcium content of approximately 0.5%) and water for a week. The ambient temperature was maintained at 22.2 ° C ± 1.7 ° C with a minimum relative humidity of 40%. The photoperiod in the room was 12 hours of light and 12 hours of darkness.

Fabric Collection in the Dosing Regimen. After a week of acclimation time (therefore, two weeks after the OVX) the daily dosing with the test compound was started. The 17α-ethynyl tradiol or test compound was administered orally, unless otherwise indicated, as a suspension in 1% carboxymethylcellulose or dissolved in 20% cyclodextrin. The animals were dosed daily for 4 days. After the dosing regimen, the animals were weighed and anesthetized with a mixture of ketamine: xylazine (2: 1, V: V) and a blood sample was collected by cardiac puncture. The animals were then sacrificed by asphyxiation with C02, the uteri were removed through an incision in the intermediate line, and the weight of a wet uterus was determined.

Cholesterol analysis. The blood samples were allowed to clot at room temperature for 2 hours, and the serum was obtained after centrifugation for 10 minutes at 3000 rpm. Cholesterol was determined using a high-precision cholesterol assay from Boehringer Mannheim Diagnostics. Briefly, the cholesterol is oxidized to colest-4-en-3-one and to hydrogen peroxide. The hydrogen peroxide is then reacted with phenol and 4-aminophenazone in the presence of peroxidase to produce a p-quinone-imine dye, which is read spectrophotometrically at 500 nm. The cholesterol concentration is then calculated against a standard curve. The complete test is automated using a Biomek Automated Workstation.

Uterine Eosinophil Peroxidase (EPO) assay. The uteri were kept at 4 ° C until the hour of the enzymatic analysis. The uteri were then homogenized in 50 volumes of 50 mM Tris buffer (pH 8.0) containing 0.005% Triton X-100. After the addition of 0.01% hydrogen peroxide and 10 mM 0-phenylenediamine (final concentrations) in Tris buffer, the increase in absorbance was checked periodically for one minute at 450 nm. The presence of eosonophils in the uterus is an indication of the estrogenic activity of a compound. The maximum speed of a second interval of 15 minutes is determined on the initial linear portion of the reaction curve.

Source of Compound: 17-α-ethynyl tradiol which was obtained from Sigma Chemical CO., Saint Louis, MO.

Hyperlipidemia: The data presented in Table 1 show the comparative results between the ovariectomized rats, the rats treated with 17-α-ethinylestradiol (EE) and the rats treated with certain compounds of this invention. Although EE2 caused a decrease in serum cholesterol when orally administered at 0.1 mg / kg / day, it also exerted a simulating effect on the uterus so that the uterine weight of EE2 was substantially greater than the uterine weight of the ovariectomized animals. This uterine response to an estrogen is well recognized in the art. Not only did the compounds of the present invention reduce serum cholesterol in comparison to ovariectomized animals, but the weight of the uterus was only minimally increased. In comparison to the estrogenic compounds known in the art, the benefit of reducing serum cholesterol without adversely affecting the weight of the uterus is unusual and desirable.

As expressed in the following data, trogenicity was also evaluated by evaluating the response of eosinophil infiltration in the uterus. The compounds of this invention did not cause a large increase in the number of eosinophils observed in the stromal layer of the uteri of ovariectomized rats. EE2 caused a substantial and expected increase in the infiltration of eosinophils. The data presented in Table 1 reflects the response of five or six rats per treatment group.

Table 1 Compound No. Dose Weight Uterine Eosinophils Cholesterol (Example No.) mg / kg% Incinerate Serum% of (Vmax) c Desc.d EE.a 0.1 227.1 * 392.7 * 71.9 * Raloxifene 0.1 75.4 * 8.4 * 60.0 * 0.01 46.0 * 4.8 -20.5 0.1 17.7 * 3.9 -2.8 1 40.6 * 3.0 14.8 Ib 0.01 3 3.9 -34.6 0.1 46.2 * 4.2 12.6 1 60.5 * 4.8 49.1 * a 17-a-ethinylestradiol b% increase in uterine weight versus controls ovariectomized c Maximum eosinophil peroxidase d Serum cholesterol decrease versus ovariectomized controls * p < .05 Osteoporosis Test Procedure Following the General Preparation Procedure described below, the rats were treated daily for 35 days (6 rats per treatment group) and sacrificed by asphyxiation with carbon dioxide on day 36. The time period of 35 days is sufficient for allow the maximum reduction in bone density, measured as described herein. At the time of sacrifice, the uteri were removed, dissected to release them from the foreign tissue, and the fluid content was expelled before determination of the dry weight, in order to confirm the estrogen deficiency associated with complete ovariectomy. The weight of the uterus was routinely reduced by approximately 75% in response to ovariectomy. The uteri were then placed in 10% neutral buffered formalin, to allow subsequent histological analysis. The right femurs were excised and digitized X-rays were generated and analyzed by an image analysis program (NIH image) in the distal metaphysis. The proximal aspect of the tibias of these animals were also explored by quantitative computed tomography. In accordance with the above procedures, the compounds of the present invention and the ethynyl tradiol (EE;) in 20% hydroxypropyl-β-cyclodextrin were orally administered to the test animals. In summary, the ovariectomy of the test animals causes a significant reduction in femur density compared to intact vehicle-treated controls. Ethinylestradiol (EE_) administered orally prevented this loss, but the risk of uterine stimulation with this treatment was still present.

The compounds of the present invention prevent bone loss in a generally dose dependent manner. Accordingly, the compounds of the present invention are useful for the treatment of post-menopausal syndrome, particularly osteoporosis.

MCF-7 Proliferation Assay MCF-7 breast adenocarcinoma cells (ATCC HTB 22) were maintained in MEM (minimal essential medium, free of phenol red, Sigma, Saint Louis, MO) supplemented with 10% fetal bovine serum (FBS) (V / V), L-glutamine (2 mM) ), sodium pyruvate (1 mM), HEPES. { (10-7 M) N-essential amino acids and bovine insulin (1 μg / ml) (maintenance medium) Ten days before the test, MCF-7 cells were changed to maintenance medium supplemented with fetal bovine serum refined with 10% dextran-coated mineral charcoal (DCC-FBS) (assay medium) instead of 10% FBS to deplete internal steroid stores The MCF-7 cells were removed from the maintenance flasks using the cell dissociation medium (Ca'VMg "free HBSS (phenol red free) supplemented with 10 M H? P? S and 2 M EDTA). cells were washed twice with the assay medium and adjusted to 80,000 cells / ml Approximately 100 ml (8,000 cells) were added to the flat bottom microculture wells (Costar 3596) and incubated at 37 ° C in an incubator Humidified with 5% CO; for 48 hours, to allow cell adhesion and balance after transfer. Serial dilutions of the drugs or DMSO were prepared as a diluent control, in the assay medium, and 50 ml were transferred to microcultures in triplicate, followed by 50 ml of assay medium for a final volume of 200 ml. After an additional period of 48 hours at 37 ° C in a humidified incubator with CO; at 5%, the microcultures were pulsed with tritiated thymidine (1 μCi / well) for 4 hours. The cultures were terminated by freezing at -70 ° C for 24 hours, followed by thawing and harvesting of the microcultures using a Skatron Semiautomatic Cell Harvester. The samples were counted by liquid scintillation using a ß Wallace BetaPlace counter.

Inhibition of mammary tumor induced by DMBA Estrogen-dependent mammary tumors were produced in female Sprague Dawley rats which were purchased from Harlan Industries, Indianapolis, Indiana. At approximately 55 days of age, the rats received a simple oral 20 mg diet of 7,12-dimethylbenz [a] anthracene (DMBA). Approximately 6 weeks after the administration of DMBA, the mammary glands were palpated at weekly intervals for the appearance of tumors. Whenever one or more tumors appeared, the longest and shortest diameters of each tumor were measured with a calibrator, measurements were recorded, and that animal was selected for experimentation. An attempt was made to uniformly distribute the various sizes of tumors in the treated and control groups, such that tumors of average size were equally distributed among the test groups. The control groups and the test groups for each experiment contained 5 to 9 animals. The compounds of foa I were administered either through intoneral injections of 2% acacia, or orally. The compounds administered orally were either dissolved or suspended in 0.2 ml of corn oil. Each treatment, which included control treatments with acacia and corn oil, was administered once a day to each test animal. After the measurement of the initial tumor and the selection of the test animals, the tumors were measured every week by the aforementioned method. The treatment and measurements of the animals continued for 3 to 5 weeks, at which time the final areas of the tumors were determined. For each treatment with compound and control, the change in the mean tumor area was determined.

Uterine Fibrosis Test Procedures Test 1 Between 3 and 20 women who had uterine fibrosis were administered with a compound of the present invention. The amount of the compound administered is 0.1 to 1000 mg / day, and the administration period is 3 months. The women were observed during the administration period, and up to 3 months after the discontinuation of the administration, for the effects on uterine fibrosis.

Test 2 The same procedure was used as in Trial 1, except that the administration period was 6 months.

Test 3 The same procedure was used as in Trial 1, except that the administration period was 1 year.

Test 4 A. Induction of fibroid tumors in guinea pigs.

Prolonged stimulation with estrogens was used to induce leiomyomata in sexually mature female guinea pigs. Animals were dosed with estradiol 3 to 5 times per week by injection for 2 to 4 months or until tumors emerged. Treatments consisting of a compound of the invention or vehicle were administered daily for 3 to 16 weeks, and then the animals were sacrificed and the uteri harvested and analyzed for tumor regression.

B. Implantation of human uterine fibrous tissue in nude mice The tissue from human leiomyomas was implanted in the peritoneal cavity and / or in the uterine myometrium of nude female, castrated, sexually mature mice. The exogenous estrogen was delivered to induce the growth of the explanted tissue. In some cases, the harvested tumor cells were cultured in vitro before implantation. The treatment consisting of a compound of the present invention or vehicle, was delivered by gastric lavage on a daily basis for 3 to 16 weeks and the implants were removed and measured for development or regression. At the time of sacrifice, the uteri were harvested to evaluate the state of the organ.

Test 5 A. Tissue from human uterine fibroid tumors was harvested and maintained in vi tro, as untransformed primary cultures. Surgical specimens were pushed through a mesh or sterile screen, or alternatively shredded to separate them from the surrounding tissue, to produce a suspension of single cells. The cells were maintained in medium containing 10% serum and antibiotic. The rates of development in the presence and absence of estrogen were determined. The cells were evaluated for their ability to produce the C3 component of the complement and its response to growth factors and growth hormone. In vitro cultures were evaluated for their proliferative response after treatment with progestin, GnRH, a compound of the present invention and vehicle. The levels of steroid hormone receptors were evaluated weekly to determine if the important characteristics of the cell were maintained, in vi tro. The tissue from 5 to 25 patients was used. The activity in at least one of the above tests indicates that the compounds of the present invention are of potential in the treatment of uterine fibrosis.

Endometriosis Test Procedure In Tests 1 and 2, the effects of administration of the compounds of the present invention at 14 and 21 days on the development of explanted endometrial tissue can be minimized.

Test 1 Twelve to thirty adult female CD strain rats were used as test animals. These are divided into three groups of equal number. The estrous cycle of all the animals was periodically verified. On the day of the proestrus, surgery was performed on each female. The females in each group had the left uterine horn excised, sectioned into small squares, and the pictures were sutured loosely at various sites adjacent to the mesenteric blood flow. In addition, the females in Group 2 had their ovaries excised. On the day after surgery, the animals in Groups 1 and 2 received intraperitoneal injections of water for 14 days, while the animals in Group 3 received intraperitoneal injections of 0.1 mg of a compound of the present invention per kilogram. body weight for the same duration. After 14 days of treatment, each female was sacrificed and the endometrial explants, the adrenal glands, the remaining uteri, and the ovaries, where applicable, were excised and prepared for histological examination. The ovaries and adrenals were weighed.

Test 2 Twelve to thirty adult female rats of the CD strain were used as the test animals. These were divided into two equal groups. The estrous cycle of all the animals was checked periodically. On the day of proestrus, surgery was performed on each of the females. The left uterine horn was removed from the females in each group, sectioned into small squares, and the boxes were loosely sutured at various sites adjacent to the mesenteric blood flow. Approximately 50 days after surgery, animals assigned to Group 1 received intraperitoneal injections of water for 21 days, while animals in Group 2 received intraperitoneal injections of 1.0 mg of a compound of the present invention per kilogram of body weight per the same duration. After 21 days of treatment, each female was sacrificed and removed and the endometrial and adrenal explants were weighed. The explants were measured as an indication of development or growth. Periodical cycles were checked periodically.

Test 3 A. Surgical induction of endometriosis Endometrial tissue autografts were used to induce endometriosis in rats and / or rabbits. The female animals at reproductive maturity underwent bilateral oophorectomy, and the estrogen was exogenously supplied thereby providing a specific and constant level of hormone. Autologous endometrial tissue was implanted in the peritoneum of 5-150 animals and estrogen was delivered to induce the development of the explanted tissue. The treatment consisting of a compound of the present invention, was delivered by gastric lavage on a daily basis for 3 to 16 weeks, and the implants were removed, and measured for development or regression. At the time of sacrifice, the intact horn of the uterus was harvested to evaluate the state of the endometrium.

B. Implantation of human endometrial tissue in nude mice The tissue originating from human endometrial lesions was implanted in the peritoneum of female nude, castrated, sexually mature mice. The exogenous estrogen was delivered to induce the development of the explanted tissue. In some cases, the harvested endometrial cells were cultured in vi tro before implantation. The treatment consisted of a compound of the present invention delivered by gastric lavage on a daily basis for 3 to 16 weeks, and the implants were removed and measured for growth or regression. At the time of sacrifice, the uteri were harvested to evaluate the state of the intact endometrium.

Test 4 A. Tissue from human endometrial lesions was harvested and maintained in vi tro as untransformed primary cultures. Surgical specimens were pushed through a mesh or sterile screen, or alternatively cut from the surrounding tissue to produce a suspension of single cells. The cells were maintained in medium containing 10% serum and antibiotic. The rates of development or growth in the presence and absence of estrogen were determined. The cells were evaluated for their ability to produce the C3 component of the complement and its response to growth factors and growth hormone. In vitro cultures were evaluated for their proliferative response after treatment with progestins, GnRH, a compound of the invention, and vehicle. The levels of steroid hormone receptors were evaluated weekly to determine if important cellular characteristics were maintained in vi tro. Tissue from 5 to 25 patients was used. The activity in any of the above assays indicates that the compounds of the present invention are useful in the treatment of endometriosis.

Inhibition of Proliferation of Aortic Smooth Muscle Cells / Restenosis Test Procedure The compounds of the present invention have the ability to inhibit the proliferation of aortal smooth muscle cells. This can be demonstrated by the use of cultured smooth muscle cells derived from rabbit aortas, proliferation being determined by measurement of DNA synthesis. Cells are obtained by the explant method as described in Ross, J. of Cell Bi o. 50: 172 (1971). The cells are seeded in 96-well microculture plates for five days. Crops become confluent and development stops. Cells are then transferred to Dulbecco's Modified Eagle's Medium (DMEM) containing 0.5-2% platelet-poor plasma, 2 mM L-glutamine, 100 U / ml penicillin, 100 mg / ml streptomycin, 1 mC / ml of 3H-thymidine, 20 ng / ml of platelet-derived growth factor, and varying concentrations of the present compounds. The stock solution of the compounds is prepared in dimethyl sulfoxide and then diluted to the appropriate concentration (0.01 -30 mM) in the above assay medium. The cells are then incubated at 37 ° C for 24 hours under 5% CO; / 95% air. At the end of 24 hours, the cells are fixed in methanol. The 3 H-thymidine incorporation in the DNA is then determined by scintillation counting as described in Bonin et al., Exp. Cell Res. 181: 475-482 (1989). The inhibition of proliferation of aortal smooth muscle cells by the compounds of the present invention is further demonstrated by the determination of their effects on exponentially growing cells. The smooth muscle cells from rabbit aortas are seeded in 12-well tissue culture plates in DMEM containing 10% fetal bovine serum, 2 mM L-glutamine, 100 U / ml penicillin, and 100 mg / ml Streptomycin After 24 hours, the cells are coupled and the medium is replaced with DMEM containing 10% serum, 2 mM L-glutamine, 100 U / ml penicillin, 100 mg / ml streptomycin, and the desired concentrations of the compounds . The cells are allowed to develop for four days. The cells are treated with trypsin and the number of cells in each culture is determined by counting using a ZM-Coulter counter. Activity in the above assays indicates that the compounds of the present invention are of potential use in the treatment of restenosis. The present invention also provides a method for alleviating post-menopausal syndrome in women, which comprises the aforementioned method using the compounds of formula I, and further comprising administering to a woman, "an effective amount of estrogen or progestin". These treatments are particularly useful for the treatment of osteoporosis and the decrease of serum cholesterol because the patient will receive the benefits of each pharmaceutical agent, while the compounds of the present invention could inhibit the undesirable side effects of estrogen and progestin. The activity of these combination treatments in any of the post-menopausal tests, mentioned below, indicates that the combination treatments are useful for relieving post-menopausal symptoms in uj. Various forms of estrogen and progestin are commercially available. Estrogen-based agents include, for example, ethinylestrogen (0.01 - 0.03 mg / day), mestranol (0.05 - 0.15 mg / day), and conjugated estrogenic hormones such as Premarin® (Wyeth-Ayerst; 0.3 - 2.5 mg / day). Progestin-based agents include, for example, medroxyprogesterone such as Provera® (Upjohn, 2.5-10 mg / day), norethylnodrel (1.0-10.0 mg / day), and nonetindrona (0.5-2.0 mg / day). A preferred estrogen-based compound is Premarin, and norethynodrel and norethindrone are preferred agents based on progestin. The method of administering each agent based on estrogen and progestin is consistent with that which is known in the art. For most of the methods of the present invention, the compounds of the formula I are administered continuously, 1 to 3 times a day. However, cyclic therapy can be especially useful in the treatment of endometriosis or it can be used acutely during painful attacks of the disease. In the case of restenosis, therapy may be limited to short intervals (1-6 months) following medical procedures such as angioplasty. As used herein, the term "effective amount" means an amount of a compound of formula I that is capable of alleviating the symptoms of the various pathological conditions described herein. The specific dose of a compound administered according to this invention, of course, will be determined by the particular circumstances surrounding the case including, for example, the compound administered, the route of administration, the health status of the patient, and the condition pathological that is treated. A typical daily dose will contain a non-toxic dose level of from about 0.1 mg to about 1000 mg / day of a compound of the present invention, and more particularly will be from about 15 mg to about 80 mg / day. As will be recognized by one of ordinary skill in the art, compounds of formula I most likely exist as amphoteri or zwitterions involving the tertiary amine structure of R3 and a monohydrogen sulfate group. The compounds of this invention also form the pharmaceutically acceptable base and acid addition salts, (for example in the R3 group, in the acid sulfate portion, or in an acid sulfate portion of an amphoteric), with a wide variety of organic and inorganic acids and bases, and include physiologically acceptable salts that are frequently used in pharmaceutical chemistry. Such salts are also part of this invention. Typical inorganic acids used to form such salts include hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, phosphoric, hypophosphoric acids and the like. Salts derived from organic acids, such as mono- and dicarboxylic aliphatic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic and hydroxyalkanedioic acids, aliphatic and aromatic sulfonic acids, can also be used. Such pharmaceutically acceptable salts include in this manner the salts of acetate, phenylacetate, trifluoroacetate, acrylate, ascorbate, benzoate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetonxybenzoate, naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate, β-hydroxybutyrate, butyne-1,4-dioate, hexin-1,4-dioate, caprate, caprylate, chloride, cinnamate, citrate, formate, fumarate, glycolate, heptanoate, hippurate, lactate, malate, maleate, hydroxyaleate, malonate, Mandelate, mesylate, nicotinate, isonicotinate, nitrate, oxalate, phthalate, terephthalate, phosphate, monoacid phosphate, diacid phosphate, metaphosphate, pyrophosphate, propiolate, propionate, phenylpropionate, salicylate, sebacate, succinate, suberate, sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate, benzenesulfonate, p-bromophenylsulfonate, chlorobenzenesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate, methanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, p-toluensu lonato, xylene sulfonate, tartrate, and the like. A preferred salt is the hydrochloride salt. The pharmaceutically acceptable acid addition salts are typically formed by the reaction of a compound of the formula I with an equimolar or excess acid amount. The reactants are generally combined in a mutual solvent such as diethyl ether or benzene. Salt is normally precipitated from the solution within approximately one hour to 10 days, and can be isolated by filtration or the solvent can be removed by conventional means. The bases commonly used for the formation of salts include ammonium hydroxide and alkali metal and alkaline earth metal hydroxides, carbonates, as well as aliphatic and primary, secondary and tertiary amines, aliphatic diamines, etc.

Bases especially useful in the preparation of the addition salts include ammonium hydroxide, potassium carbonate, methylamine, diethylamine, ethylenediamine and cyclohexylamine. The pharmaceutically acceptable base addition salts are typically formed by the reaction of a compound of formula I with an equimolar or excess amount of base. The reagents are generally combined in a mutual solvent such as diethyl ether, ethyl acetate, alcohols or benzene. The salt usually precipitates from the solution within about one hour to 10 days and can be isolated by filtration or the solvent can be removed by conventional means. Compounds having two sulfate moieties can be converted to a salt by addition of base and an amphoteric salt by reaction of the amphoteric with one equivalent of base. The reagents are generally combined in a mutual solvent such as diethyl ether, ethyl acetate, alcohols or benzene. The salt normally precipitates from the solution within about an hour to 10 days and can be isolated by filtration or the solvent can be removed by conventional means. Pharmaceutically acceptable salts generally have improved solubility characteristics compared to the compound from which they are derived, and are thus often more suitable for formulation as liquids or emulsions. It is usually preferred to administer a compound of the formula I in the form of a salt by the addition of acid, as is customary in the administration of pharmaceutical products having a basic group, such as e? piperidino ring. It is also advantageous to administer such compound by the oral route. For these purposes the following oral dosage forms are available. The compounds of this invention can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal. These compounds are preferably formulated before administration, the selection of which will be decided by the attending physician. Thus, yet another aspect of the present invention is a pharmaceutical composition comprising an effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof, optionally containing an effective amount of estrogen or progestin, and a carrier , pharmaceutically acceptable diluent or excipient. The total active ingredients in such formulations comprise from 0.1% to 99.9% by weight of the formulation. By "pharmaceutically acceptable" it is meant that the carrier, diluent, excipient and salt, must be compatible with the other ingredients of the formulation, and not harmful to the container thereof. The pharmaceutical formulations of the present invention can be prepared by methods known in the art using well known and readily available ingredients. For example, compounds of formula I with or without an estrogen or progestin compound can be formulated with common excipients, diluents, or carriers, and formed into tablets, capsules, suspensions, powders, and the like. Examples of excipients, diluents, and carriers that are suitable for such formulations include the following: fillers and extenders such as starch, sugars, mannitol, and silicic derivatives; binding agents such as carboxymethylcellulose and other cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone; wetting agents such as glycerol; disintegrating agents such as calcium carbonate and sodium hydrogen carbonate; agents for delaying dissolution such as paraffin; resorption accelerators such as quaternary ammonium compounds; surface active agents such as cetyl alcohol, glycerol monostearate; adsorptive carriers, such as kaolin and bentonite; and lubricants such as talc, calcium and magnesium stearate, and solid polyethylene glycols. The compounds may also be formulated as elixirs or solutions for "convenient oral administration or as appropriate solutions for parenteral administration, for example, by intramuscular, subcutaneous or intravenous routes." In addition, the compounds are well suited for formulation as forms of administration. sustained release dose and the like The formulations can be constituted in such a way that they release the active ingredient uniquely or preferably at a particular physiological site, possibly over a period of time.The coatings, wraps, and • Protective matrices can be made, for example, from polymeric substances or waxes.

The compounds of the formula I, alone or in combination with a pharmaceutical agent of the present invention, will generally be administered in a convenient formulation. The following formulation examples are illustrative only and are not intended to limit the scope of the present invention.

Formulations In the following formulations, "active ingredient" means a compound of formula I, or a salt or solvate thereof.

Formulation 1: Gelatin Capsules Hard gelatin capsules are prepared using the following: Ingredient Quantity (mg / capsule) Active ingredient 0.1 - 1000 Starch, NF 0 - 650 Flowable powder starch 0 - 650 350 centistoke silicone fluid 0 - 15 The above formulation can be changed in compliance with the reasonable variations provided. A tablet formulation is prepared using the following ingredients: Formulation 2: Tablets Ingredient Quantity (mg / tablet) Active ingredient 2.5 - 1000 Microcrystalline cellulose 200 - 650 Silicon dioxide, smoked 10 - 650 Stearic acid 5 - 15 The components are mixed and compressed to form tablets. Alternatively, the tablets each containing 2.5-1000 mg of active ingredient are constituted as follows: Formulation 3: Tablets Ingredient Amount (mg / tablet; Active ingredient 25 - 1000 Starch 45 Microcrystalline cellulose 35 Polyvinylpyrrolidone (as a 4% solution in water) Sodium carboxymethylcellulose 4.5 Magnesium stearate 0.5 Talcum 1 The active ingredient, starch, and cellulose are passed through a No. 45 mesh American sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a No. 14 American mesh screen. The granules thus produced are dried at 50 ° C-60 ° C and passed through a US No. 18 mesh sieve. Sodium carboxymethylcellulose, magnesium stearate, and talcum, previously passed through a No. 60 mesh American sieve, are then added to the granules, after mixing, they are compressed into a tablet-forming machine, to produce tablets. The suspensions each containing 0.1 to 1000 mg of medication per 5 ml of dose, are elaborated as follows: Formulation 4: Suspensions Ingredient Quantity (mg / 5 ml) Active ingredient 0.1 - 1000 mg Sodium carboxymethylcellulose 50 mg Syrup 1.25 mg Benzoic acid solution 0.10 ml Flavoring q.s. Color c.s. Purified water up to 5 ml The drug is passed through a No. 45 mesh American sieve and mixed with the sodium carboxymethylcellulose and the syrup to form a smooth paste. The benzoic acid solution, the flavor and the color are diluted with some of the water and added, with stirring. Sufficient water is then added to produce the required volume. An aerosol solution is prepared containing the following ingredients: Formulation 5: Aerosol Ingredient Quantity (% by weight) Active ingredient 0.25 Ethanol 25.75 Propellant 22 (chlorodifluoromethane) 70.00 The active ingredient is mixed with ethanol and the mixture is added to a portion of propellant 22, cooled to 30 ° C, and transferred to a filling device. The required amount is then fed to a stainless steel vessel and diluted with the remaining propellant. The valve units are then adjusted to the container. Suppositories are prepared as follows: Formulation 6: Suppositories Ingredient Quantity (mg / suppository) Active ingredient 250 Saturated fatty acid glycerides 2,000 The active ingredient is passed through a No. 60 American mesh screen and suspended in the saturated fatty acid glycerides previously melted, using the minimum necessary heat. The mixture is then emptied into a suppository mold with a nominal capacity of 2 grams and it is to be cooled. An intravenous formulation is prepared as follows: Formulation 7: Intravenous solution Ingredient Quantity Active ingredient 50 Isotonic saline 1,000 ml The solution of the above ingredients is administered intravenously to a patient at a rate of approximately 1 ml per minute Formulation 8: Capsule in Combination I Ingredient Quantity (mg / capsule; Active ingredient 50 Premarin 1 Avicel pH 101 50 Starch 1500 117.50 Silicone oil 2 Tween 80 0.50 Cab-O-Sil 0.25 Formulation 9: Capsule in Combination II Ingredient Quantity (mg / capsule; Active ingredient 50 Norethylnodrel 5 Avicel pH 101 82.50 Starch 1500 90 Silicone oil 2 Tween 80 0.50 Formulation 10: Tablet in Combination Ingredient Quantity (mg / capsule) Active ingredient 50 Premarin 1 Corn starch NF 50 Povidone, K29-32 6 Avicel pH 101 41.50 Avicel pH 102 136.50 Crospovidone XL10 2.50 Magnesium stearate 0.50 Cab-O-Sil 0.50 It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects to which it relates.

Having described the invention as above, property is claimed as contained in the following:

Claims (18)

1. A compound of the formula I (I) characterized in that R1 is hydrogen, hydroxyl, alkoxy of 1 to 4 carbon atoms, -OCOO (alkyl of 1 to 6 carbon atoms), -OCO (alkyl of 1 to 6 carbon atoms), -OCOAr wherein Ar is phenyl or optionally substituted phenyl, -OSO; (straight chain alkyl of 4 to 6 carbon atoms), or -OS03H; R2 is R1, Cl or F; with the proviso that at least R1 or R2 is -OSO3H; R3 is 1-piperidinyl, 1-pyrrolidinyl, methyl-1-pyrrolidinyl, dimethyl-1-pyrrolidino, 4-morpholino, dimethylamino, diethylamino, diisopropylamino, or 1-hexamethyleneimino; and n is 2 or 3; or a pharmaceutically acceptable salt or solvate thereof.

2. A compound according to claim 1, characterized in that R1 and R2 are each -OSO, H.

3. A compound according to claim 1, characterized in that R is hydroxyl and R "is -OS03H.

4. A compound according to claim 1, characterized in that R2 is hydroxyl and R1 is -OS03H.

5. A compound according to claim 1, characterized in that n = 2.

6. A compound according to claim 1, characterized in that R3 is 1-piperidinyl.

7. A pharmaceutical composition, characterized in that it comprises a compound according to claim 1, or a pharmaceutically acceptable salt thereof and, optionally, an effective amount of estrogen or progestin, in combination with a pharmaceutically acceptable carrier, diluent or excipient.

8. The use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for alleviating the symptoms of post-menopausal syndrome in a woman.

9. The use according to claim 8, characterized in that the pathological condition of the post-lessus syndrome is osteoporosis.

10. The use according to claim 8, characterized in that the pathological condition of the post-menopausal syndrome is related to a cardiovascular disease.

11. The use according to claim 10, characterized in that the cardiovascular disease is hyperlipidemia.

12. The use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for inhibiting estrogen-dependent cancer in a patient.

13. The use "of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for inhibiting uterine fibroid disease in a woman.

14. The use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for inhibiting endometriosis in a woman.

15. The use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for inhibiting the proliferation of aortal smooth muscle cells in a human.

16. The use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for inhibiting restenosis in a human.

17. The use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, and estrogen for the preparation of a medicament for alleviating the symptoms of post-menopausal syndrome in a woman.

18. The use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, and progestin for the preparation of a medicament for alleviating the symptoms of post-menopausal syndrome in a woman. SUMMARY OF THE INVENCIÓ The invention provides the compounds of the formula I, wherein R 1 is hydrogen, hydroxyl, alkoxy of 1 to 4 carbon atoms, -OCOO (alkyl of 1 to 6 carbon atoms), -OCO (alkyl of 1 to 6 carbon atoms) carbon), -OCOAr wherein Ar is .phenyl or optionally substituted phenyl, -OSO; (straight chain alkyl of 4 to 6 carbon atoms), or -OS03H; R "is R1, Cl or F with the proviso that at least one of R1 or R2 is -0S03H, R3 is 1-piperidinyl, 1-pyrrolidinyl, methyl-1-pyrrolidinyl, dimethyl-1-pyrrolidino, 4-morpholino , dimethylamino, diethylamino, diisopropyl, or 1-hexamethyleneimino, and n is 2 or 3, or a pharmaceutically acceptable salt or solvate thereof The present invention further relates to pharmaceutical compositions containing the compounds of the formula (I), which optionally contain estrogen or progestin, and the use of such compounds, alone, or in combination with estrogen or progestin, to alleviate the symptoms of post-menopausal syndrome, particularly osteoporosis, pathological conditions related to the cardiovascular system, and estrogen-dependent cancer The compounds of the present invention are also useful for inhibiting uterine fibroid disease and endometriosis in women and the proliferation of aortal smooth muscle cells, particularly restenosis. , in humans.