MXPA97009467A - Pharmaceutical compositions to minimize loss or - Google Patents

Abstract

The present invention provides a method for minimizing the effect of the bone loss of a compound of the formula (II) or a pharmaceutically acceptable salt thereof, wherein the compound of the formula (II) is administered to a mammal in need of treatment, comprising the concurrent or sequential administration to the mammal of an effective amount of a compound of the formula (I), wherein each R1 is independently -H, -OH, -O (alkyl of 1 to 4 carbon atoms), -OCOC6H5 , -OCO (alkyl of 1 to 6 carbon atoms), or -OSO 2 (alkyl of 4 to 6 carbon atoms), and R 2 is 1-piperidinyl, 1-pyrrolidinyl, methyl-1-pyrrolidinyl, dimethyl-1-pyrrolidinyl, -morpholino, dimethylamino, diethylamino or 1-hexamethyleneimino, or a pharmaceutically acceptable salt thereof. The method is also provided for minimizing the bone loss induced by the administration of a compound of the formula (II), which comprises administering an anabolic-bone agent concurrently or sequentially. Pharmaceutical compositions are also provided

Description

ANTAGONISTS OF CALCIUM CHANNELS DESCRIPTION OF THE INVENTION This invention relates to the discovery that a group of 2-aryl-3-aroylbenzo [b] thiophenes are antagonists of calcium channels in vascular tissue. Estrogen replacement therapy is generally recognized as a producer of beneficial effects on the cardiovascular system in postmenopausal women. See Knopt, Obs tet. Gynecol. , 12, 23s-30s (1988). In postmenopausal women who receive estrogen, the proportion of cardiovascular mortality is reduced by approximately 30% to approximately 50%, and the proportion of cerebrovascular mortality is reduced by approximately 50%. See Stampfer et al., N. Engl. J. Med. , 325, 756-762 (1991). Although these beneficial cardiovascular effects may involve alterations in the profile of the lipids, recent data suggest that estrogen may also have beneficial effects on the vascular responses of the atherosclerotic coronary arteries. See Gisclard et al., J.

REF: 26325 Pharmacol. and Experimental Therapeuti cs, 244, 19-22 (1988); Williams et al., Circulate on, 81, 1680-1687 (1990); Gangar et al., Lancet, 388, 839-842 (1991); and Williams et al., JACC, 20, 452-457 (1992). The endothelium-independent and endothelium-dependent effects of estrogen have been described in vascular tissue. See Jiang et al., Br. J. Pharmacol. , 104, 1033-1037 (1991); Jiang et al., Ameri can Journal of Physi olgy, 32, H271-H275 (1992); Cheng and Gruetter, European Journal of Pharmacol. , 215, 171-176 (1992); Mügge et al., Cardi ovas. Res. , 21, 1939-1942 (1993); Rooms and collaborators, European Journal of Pharmacol. , 258, 47-55 (1994); Williams et al., Circul ation, 81, 1680-1687 (1990); Cheng et al., Life Sci ences, 10, 187-191 (1994); Gilligan et al., Circulation, 89, 2545-2551 (1994); and Reis et al., Circul a ti on, 89, 52-60 (1994). Several reports have also suggested that the vasodilatory effects of estradiol and / or its ability to attenuate contractile responses can be mediated by the inhibition of calcium influx via voltage-dependent calcium channels. See Jiang et al., Br. J. Pharmacol. , 104, 1033-1037 (1991); Jiang et al., Ameri can Journal of Physi olgy, 32, H271-H275 (1992); Collins et al., Lancet, 341, 1264 (1993); Muck et al., Med. Sci. Res. , 22, 19 (1994); and Salas et al., European Journal of Pharmacol. , 258, 47-55 (1994). Others have postulated that estradiol may increase the cyclic AMP or cyclic GMP content, or increase the ATP-sensitive potassium channels. See Mügge et al., Cardiovas. Res. , 21, 1939-1942 (1993). The 2-aryl-3-aroylbenzo [b] thiophene compounds that are used in the methods of this invention were first developed by Jones and Suarez as anti-fertility agents. See U.S. Patent No. 4,133,814 (issued January 9, 1979). These compounds are generally useful for suppressing the growth of mammary tumors. Jones later found that a group of these compounds are particularly useful for antiestrogenic and antiandrogenic therapy, especially in the treatment of mammary and prostatic tumors. See U.S. Patent No. 4,418,068 (issued November 29, 1983). One of these compounds, 6-hydroxy-2- (4-hydroxyphenyl) -3- [4- (2-piperidino-ethoxy) benzoyl] benzo [b] thiophene was clinically studied for the treatment of breast cancer.

This compound is called raloxifene, formerly keoxifene. This invention provides methods for antagonizing or blocking calcium channels in vascular tissue, comprising administering to a warm-blooded animal in need thereof, an effective amount of a compound of the formula R I) wherein R1 and R3 are independently hydrogen, alkyl of 1 to 4 carbon atoms, -CO- (alkyl of 1 to 6 carbon atoms), -CH2Ar, or -COAr, wherein Ar is phenyl or substituted phenyl; R2 is selected from the group consisting of pyrrolidino, hexamethyleneimino, and piperidino; or a pharmaceutically acceptable salt thereof. The present invention also provides the use of the compounds of formula I, or pharmaceutically acceptable salts thereof, for the manufacture of a medicament for antagonizing or blocking calcium channels in vascular tissue. Figure 1: Contractile responses to norepinephrine (upper part), serotonin (intermediate part), and U46619 (lower part) in rat aortic rings in the presence and absence of endothelium. The integrity of the endothelium was measured by the challenge with acetylcholine as indicated in the methods. The points are the average values and the vertical bars represent the standard error of the mean for the number of rings indicated in the parentheses. Figure 2: Effect of 16 on the contractile response to norepinephrine (upper part), serotonin (intermediate part), and U46619 (lower part) in the rat aorta that has an intact endothelium. The points are the average values and the vertical bars represent the standard error of the mean for the number of rings indicated in parentheses. Figure 3: Effect of 16 on the contractile response to norepinephrine (upper part), serotonin (intermediate part), and U46619 (lower part) in rat aortic rings without an intact endothelium. The points are the average values and the vertical bars represent the standard error of the mean for the number of rings indicated in parentheses. Figure 4: Effect of 14 on the contractile response to serotonin in rat aorta with (top) and without (bottom) an intact endothelium. The points are the average values and the vertical bars represent the standard error of the mean for the number of rings indicated in parentheses. Figure 5: Effect of 14 on the contractile response to norepinephrine in rat aortic rings with (top) and without (bottom) an intact endothelium. The points are the average values and the vertical bars represent the standard error of the mean for the number of rings indicated in parentheses. Figure 6: Effect of β-estradiol (upper part) and 16 (lower part) on the contractile response of the rat aorta that has an intact endothelium for BayK 8644. The points are the mean values and the vertical bars represent the standard error of the mean for the number of tissues indicated in parentheses. Figure 7: Effect of β-estradiol (upper part) and 16 (lower part) on the contractile response on the rat aorta without an intact endothelium for BayK 8644. The points are the mean values and the vertical bars represent the standard error of the mean for the number of tissues indicated in parentheses. Figure 8: Effect of 14 (10 ~ 6 and 107M) on the contractile response to BayK 8644 in rat aorta lacking an endothelium. The points are the average values and the vertical bars represent the standard error of the mean for the number of tissues indicated in parentheses. The present invention relates to the discovery that a select group of 2-aryl-3-aorylbenzo [b] thiophenes (benzo [b] thiophenes), the compounds of the formula I, are calcium channel antagonists. Therefore, the present invention provides methods for antagonizing or blocking calcium channels in vascular tissue. One aspect of the invention is a method for treating cardiac disorders, including but not limited to variant angina, angina induced by physical exercise, unstable angina, myocardial damage by ischemia-reperfusion, and arrhythmias. Another aspect is a method to treat cerebrovascular disorders, including but not limited to cerebral vasospasm due to arterial rupture, shock and migrainous headaches. Another aspect is a method to treat kidney disorders by increasing renal clearance due to increases in renal blood flow, useful to reduce renal failure. Another aspect is to treat gastrointestinal disorders, including but not limited to diseases related to diarrhea, such as IBS and IBD, predominant diarrhea. Another aspect is a method to treat hypertension. Therapeutic treatments provided by this invention are practiced by administering to a warm-blooded animal in need thereof, a pharmaceutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof. In the above formula, the term "alkyl of 1 to 6 carbon atoms" represents a linear, cyclic, or branched alkyl chain having from 1 to 6 carbon atoms. Typical alkyl groups of 1 to 6 carbon atoms include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, n-hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. The term "alkyl of 1 to 4 carbon atoms" represents a linear or branched alkyl chain having from 1 to 4 carbon atoms. Typical alkyl groups of 1 to 4 carbon atoms include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, and t-butyl. The term "Ar" represents groups such as phenyl and substituted phenyl. The term "substituted phenyl" as used herein, represents a phenyl group substituted with one or more portions selected from the group consisting of halogen, hydroxyl, cyano, nitro, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, acetyl, formyl, trichloromethyl, or trifluoromethyl. Examples of a substituted phenyl group include 4-chlorophenyl, 2,6-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-bromophenyl, 3,4-dibromophenyl, 3- chloro-4-fluorophenyl, 2-fluorophenyl, 4-hydroxyphenyl, 3-hydroxyphenyl, 2,4-dihydroxyphenyl, 3-nitrophenyl, 4-nitrophenyl, 4-cyanophenyl, 4-methylphenyl, 4-ethylphenyl, 4-methoxyphenyl, 4- propylphenyl, 4-n-butylphenyl, 4-t-butylphenyl, 3-fluoro-2-methylphenyl, 2,3-difluorophenyl, 2,6-difluorophenyl, 2,6-dimethylphenyl, 2-fluoro-5-methylphenyl, 2, 4,6-trifluorophenyl, 2-trifluoromethylphenyl, 2-chloro-5-trifluoromethylphenyl, 3,5-bis (trifluoromethyl) phenyl, 2-methoxyphenyl, 3-methoxyphenyl, 3,5-dimethoxyphenyl, 4-hydroxy-3-methylphenyl, 3, 5-dimethyl-4-hydroxyphenyl, 2-methyl-4-nitrophenyl, 4-methoxy-2-nitrophenyl, 2,4-dinitrophenyl, and the like. The term "C 1 -C 4 alkoxy" represents groups such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, and the like. The term "halogen" represents fluoro, chloro, bromo and iodo. The term "blocking" or "antagonizing" indicates that the compounds of formula I bind to the calcium channels in the vascular tissue and thereby inhibit the flow of calcium. The term "pharmaceutically effective amount" is used herein to represent an amount of the compound of formula I, which is capable of antagonizing or blocking calcium channels in vascular tissue. The particular dose of the compound of formula I will of course be determined by the particular circumstances surrounding the case, including the compound administered, the route of administration, the particular condition treated, and similar considerations.

The term "warm-blooded animal", as used herein, includes humans; companion animals, such as dogs and cats; and domestic animals, such as horses, cattle, sheep, pigs, goats and chickens. Preferably, the warm-blooded animal is a human or companion animal. More preferably, the warm-blooded animal is a human. While all compounds of formula I are useful for antagonizing or blocking calcium channels in vascular tissue, certain compounds are preferred. Preferably, R1 and R3 are independently hydrogen, alkyl of 1 to 4 carbon atoms, -CO- (alkyl of 1 to 6 carbon atoms), or benzyl, and R2 is piperidino or pyrrolidino.

Compounds representative of this preferred group include 6-hydroxy-2- (4-hydroxyphenyl) -3- [4- (2-pyrrolidinoethoxy) benzoyl] -benzo [b] thiophene, 6-methoxy-2- (4-methoxyphenyl) -3- [4- (2-piperidinoethoxy) benzoyl] benzo [b] thiophene, 6-acetoxy-2- (4-acetoxyphenyl) -3- [4- (2-pyrrolidinoethoxy) benzoyl] -benzo [b] thiophene, and 6-benzyloxy-2- (4-benzyloxyphenyl) -3- [4- (2-piperidinoethoxy) benzoyl] benzoyl] -benzo [b] thiophene.

More preferably, R1 and R3 are independently hydrogen or alkyl of 1 to 4 carbon atoms, and R2 is piperidino or pyrrolidino. Representative compounds from this most preferred group include 6-hydroxy-2- (4-hydroxyphenyl) -3- [4- (2-pyrrolidinoethoxy) benzoyl] benzo [b] thiophene, 6-hydroxy-2- (4-hydroxyphenyl) ) -3- [4- (2-piperidinoethoxy) benzoyl] benzo [b] thiophene, 6-methoxy-2- (4-methoxyphenyl) -3- [4- (2-pyrrolidinoethoxy) -benzoyl] benzo [b] thiophene , and 6-methoxy-2- (4-methoxyphenyl) -3- [4- (2-piperidinoethoxy) benzoyl] benzo [b] thiophene. more preferably, R1 and R3 are hydrogen and R2 is pyrrolidino. This most preferred compound is 6-hydroxy-2- (4-hydroxyphenyl) -3- [4- (2-pyrrolidinoetho) benzoyl] -benzo [b] thiophene. The compounds of the formula I used in the methods of the present invention can be made according to established procedures, such as those described in U.S. Patent Nos. 4,133,814, 4,418,068, and 4,380,635, all of which are incorporated by reference into US Pat. the present. In general, the process begins with β-hydroxy-2- (4-hydroxyphenyl) benzo [b] thiophene. this initial compound is protected, acylated at carbon 3 with a 4- (2-aminoethoxy) enzoyl group, and optionally deprotected to form the compounds of formula I. Examples of the preparation of such compounds are given in the North American patents discussed previously. The compounds used in the methods of this invention form pharmaceutically acceptable acid addition salts and, wherein R 1 and / or R 3 is hydrogen, pharmaceutically acceptable base addition salts, with a wide variety of organic and inorganic acids and bases, including the physiologically acceptable salts which are frequently used in pharmaceutical chemistry. 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, aromatic 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-acetoxybenzoate, naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate, and b-hydroxybutyrate, butyn-1,4-dioate, hexin-1,6-dioate, caprate, caprylate, chloride, cinnamate, citrate, formate, fumarate, glycolate, heptanoate, decanoate, hippurate, lactate, malate, maleate, hydroxyaleate malonate, mandelato, mesylate, nicotinate, isonicotinate, nitrate, oxalate, phthalate, terephthalate, phosphate, monoacid phosphate diacid, metaphosphate, pyrophosphate, propiolate, propionate, phenylpropionate, salicylate, sebacate, succinate, suberate, sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate, benzenesulfonate, p-bro-ofhenylsulfonate, chlorobenzenesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate, methanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, p-toluenesulfonate, xylene sulfonate, tartrate and the like. The most 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 amount of the acid. The reactants are generally combined in an organic solvent such as methanol, diethyl ether or benzene. The salt normally precipitates from the solution within about an hour to 10 days, and can then be isolated by filtration, or the solvent can be removed by conventional means. Bases commonly used for the formation of salts include ammonium hydroxide and alkali metal and alkaline earth metal hydroxides, carbonates, as well as primary, secondary or tertiary aliphatic amines, and aliphatic diamines. Bases especially useful in the preparation of the addition salts include ammonium hydroxide, potassium carbonate, methylamine, diethylamine, ethylenediamine, and cyclohexylamine. These salts are generally prepared by the reaction of a compound of the formula I, wherein R1 and / or R3 are hydrogen, with one of the above bases in an organic solvent, such as methanol, diethyl ether or benzene. The salts are isolated as described in the preceding paragraph. These 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. The compounds of the formula I are preferably formulated prior to administration such as in a pharmaceutical formulation comprising a compound of the formula I and a pharmaceutically acceptable carrier, diluent or excipient. These pharmaceutical formulations are prepared by known procedures using well-known and readily known ingredients. In making these compositions, the active ingredient will usually be mixed with a carrier, diluted with a carrier, or enclosed within a carrier, which may be in the form of a capsule, sachet, paper or other container. When the carrier serves as a diluent, it can be a solid, semi-solid or liquid material which acts as a vehicle, excipient or medium for the active ingredient. The compositions may be in the form of tablets, lozenges, powders, pills, sacks, capsules, elixirs, suspensions, emulsions, solutions, syrups, sprays, ointments containing, for example, up to 10% by weight of the active compound, gelatin capsules soft and hard, skin patches, suppositories, sterile injectable solutions, and sterile packaged powders. Some examples of suitable carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum, acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, crosslinked polyvinylpyrrolidone, cellulose or derivatives thereof, water syrup, methyl cellulose, methyl and propyl hydroxybenzoates, talc, magnesium stearate and mineral oil. The formulations may also include lubricating agents, wetting agents (e.g. surfactant), emulsifying and suspending agents, disintegrating agents, preservatives, sweetening agents, or flavoring agents. The compositions of the invention can be formulated to provide quick releasesustained or delayed active ingredient after administration to the patient, by employing procedures well known in the art. The particular dose of a compound of the formula I, required to antagonize or block the calcium channels in the vascular tissue, according to the invention, will depend on the severity of the condition, the route of administration, and the related factors. which will be decided by the attending physician. In general, the effective daily doses will be from about 0.1 to about 1000 mg / day, and more typically from about 50 to about 250 mg / day. Such doses will be administered to a subject in need thereof from one to about three times per day, or more frequently as necessary to effectively treat the condition or symptom. It is usually preferred to administer a compound of the formula I in the form of an acid addition salt, as is customary in the administration of pharmaceutical products that possess a basic group, such as the piperidino group. For these purposes the following oral dosage forms are available.

In the following formulations, "Active Ingredient" means a compound of formula I.

Formulation 1: Gelatin Capsules Hard gelatin capsules are prepared using the following ingredients: Ingredient Quantity (mg / capsule) Active ingredient 0.1 - 1000 Starch, NF 0 - 650 Fluid starch powder 0 - 650 Fluid silicone 350 centistokes 0 - 15 The ingredients are mixed, passed through a No. 45 mesh US sieve, and filled into hard gelatin capsules.

Examples of raloxifene-specific capsule formulations that have been made include those shown below: Formulation 2: Raloxifene Capsule Ingredient Quantity (mg / capsule; Raloxifene 1 Starch, NF 112 Flowable powder starch 225.3 350 centistokes fluid silicone 1.7 Formulation 3: Raloxifene capsule Ingredient Quantity (mg / capsule) Raloxifene 5 Starch, NF 108 Fluid starch powder 225. 3 Fluid silicone 350 centistokes 1. 7 Formulation 4: Raloxifene capsule Ingredient Quantity (mg / capsule) Raloxifene 10 Starch, NF 103 Fluid starch powder 225. 3 Fluidic Silicone of 350 centistokes 1. 7 Formulation 5: Raloxi capsule pheno Ingredient Quantity (mg / cápsula¡ Raloxifene 50 Starch, NF 150 Starch flowable powder 397 Silicone fluid 350 centistokes 3 of. 0 The specific formulations above may be changed in compliance with the reasonable variations provided.

A tablet formulation is prepared using the following ingredients: Formulation 6: Tablets Ingredient Quantity (mg / tablet) Active ingredient 0.1 1000 Cellulose, microcrystalline 0 650 Silicon dioxide, smoked 0 650 Stearic acid 0 15 The components are mixed and compressed to form tablets.

Alternatively, the tablets each containing 0.1 to 1000 mg of active ingredient, are constituted as follows: Formulation 7: Tablets Ingredient Quantity (mg / tablet) Active ingredient 0.1 - 1000 Starch 45 Cellulose, microcrystalline 35 Polyvinylpyrrolidone 4 (as a 10% 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 sieve of No. 14 mesh US The granules so produced are dried at 50 ° -60 ° C and passed through a sieve U.S. No. 18 mesh the sodium carboxymethylcellulose, magnesium stearate, and talc, previously passed through a sieve of No. 60 mesh US, they are then added to the granules which, after mixing, are compressed in a tablet-forming machine, to produce tablets. The suspensions each containing 0.1-1000 mg of active ingredient per 5 ml of dose, are constituted as follows: Formulation 8: 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 c.s. Color c.s. Purified water cbp 5 ml The active ingredient is passed through a No. 45 mesh US sieve and mixed with the sodium carboxymethyl cellulose 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.

Illustrative compounds that can be used in the methods of the present invention are shown in Table 1.

Table 1 Compound No. R1 and R3 R¿ Form 1 - . 1 -C (0)? (~ -F piperidino base 2 - . 2 -C (0,? O ^ -F piperidino HCl 3 - . 3 -C (O) - / piperidino base 4 - . 4 -C (?) Piperidino HCl - . 5 -C (0) CH2CH2CH3 piperidino base 6 -. 6 -C (0) CH2CH2CH3 piperidino HCl 7 -. 7 -C (0) C (CH3) 3 piperidino base 8 -. 8 -C (0) C (CH3) 3 piperidino HCl 9 -. 9 -C (0) CH2C (CH3) 3 piperidino base -. 10 -C (0) CH2C (CH3) 3 piperidino HCl eleven - . 11 -C (O) CH3 piperidino HCl 12 piperidino base -C (0, -? \) 13 H piperidino base 14 H piperidino HCl Table 1 Compound No. R1 and R3 R2 Shape H pyrrolodino base 16 H pyrrolodino HCl 17 H hexamethyleneimine HC1 18 CH3 piperidino HCl The utility of the compounds of the formula I is illustrated by the positive impact they have in at least one of the experiments described below: Methods Male Wistar rats (250-350 g, Charles River Laboratories, Portage, MI) were sacrificed by cervical dislocation. The aorta was removed and cleared of foreign tissue and cut into ring segments, each approximately 4-5 mm in length. In some tissues, the endothelium was removed by rotating the ring segment around the tip of a forceps 10 times. The lack of tissue relaxation to acetylcholine (10 ~ 6M) after contraction with norepinephrine (10 ~ 7M) was used as evidence for bare endothelium. Tissues with an intact endothelium were relaxed with acetylcholine with a relaxation of 88.5 ± 1.6% (n = ll).

The ring segments were placed between two stainless steel hooks and mounted in baths for isolated organs, containing 10 ml of Krebs bicarbonate buffer, modified from the following composition (mM): NaCl, 118.2; KCl, 4.6 'CaCl2 • 2H20, 1.6; KH2P04, 1.2; MgSO4, 1.2; glucose, 10; and NaHCO 3, 24.8. The solutions for organ bath were aerated with 95% 02/5% C02 and maintained at 37 ° C. The tissues were placed under an optimal force of 4 grams and balanced for one hour with washes every 15 minutes. Changes in force were recorded and analyzed by a Biopac MP100 data acquisition system (World Precision Instruments, Sarasota, FL) via Sensotec transducers (model MBL 5514-02) (Sensotec Inc. Columbus, OH). All tissues were initially challenged with KCl (67 mM) to establish viability. The concentration-response curves were generated in a cumulative manner and are reported as a percentage of an initial contraction by KCl (67 mM) produced in each tissue. The vehicle, ß-estradiol, 14 or 16 was added to the tissues 60 minutes before the start of the concentration-response curves for the agonists. In the BayK 8644 studies, the tissues were exposed to 10 mM KCl before initiating the concentration-response curves for BayK 8644. Only one concentration-response curve was generated for the agonist in each tissue. All results were expressed as the mean ± SE, where n represents the number of rings examined.

Chemicals 5-HT, acetylcholine, norepinephrine, U46619, and ß-estradiol were purchased from Sigma Chemical Company (Saint Louis, MO). BayK 8644 was purchased from Research Biochemicals Inc. (Wayland, MA). Diltiazem, nifedipine, nitrendipine, and compounds Nos. 14 and 16 (hereinafter referred to as 14 and 16) were synthesized at Lilly Research Laboratories (Indianapolis, IN).

RESULTS The contractile response of rat aorta to norepinephrine, serotonin and thromboxane mimic U46619 was clearly endothelial dependent (Figure 1). For each agonist, the contractile response was greater in the absence of endothelium, suggesting that the contractile response was modulated by the relaxing agonist (s) released from the endothelium. The non-steroidal benzothiophene derivative 16 (10"d and 10" 5 M) potently inhibited the contractile responses to norepinephrine, serotonin and U46619 in rat aorta, and the inhibition of the contractile response appeared independent of the endothelium (Figures 2 and 3). ). As previously observed with ß-estradiol, 16 was more potent for inhibiting the contractile response to serotonin relative to norepinephrine or U46619. As with 16, 14 it also inhibited the contractile responses to serotonin (Figure 4), an inhibition that was greater than with norepinephrine (Figure 5); effects that were independent of an intact endothelium. It is known that serotonin-induced contraction utilizes extracellular calcium via voltage-dependent calcium channels, while vascular contraction for norepinephrine is more highly dependent on the activation of phosphoinositide production and the use of intracellular calcium stores . In this way, serotonin contractions are more sensitive to inhibition by calcium channel blockers than norepinephrine contractions.

Cohen and Berkowitz, Bl ood Vessel s, 13, 139-154 (1976); and Gouw et al., Naunyn -Schmi edebergf s Arch.

Pharmacol. , 339, 533-539 (1989). Because 14 and 16 appeared to be most effective in inhibiting the contractile responses induced by serotonin, 14 and 16 may be acting as a calcium channel antagonist. For this reason, the effectiveness of 14 and 16 to inhibit the contractile responses induced by the calcium agonist BayK 8644 was examined. See Brown et al., Na t ure, 311, 570-572 (1984). In the absence of an intact endothelium the inhibition of contraction for BayK 8644 occurred with 14 and 16 (Figures 7 and 8). This data with BayK 8644 document the inhibition of the contractile response for this calcium agonist by 14 and 16.

Discussion The ability of estrogen to block calcium channels can contribute to the beneficial effects of estrogen, when used as replacement therapy in postmenopausal women. It is known that estrogen possesses calcium channel antagonist activity, an effect that has been observed in rabbit and pig coronary arteries. Recently, several groups have been tried to identify compounds that can mimic the beneficial hemodynamic effects of estrogen, while minimizing the uterotropic or undesirable effects of estrogen when used as replacement therapy. In this regard, a series of partial agonist compounds of the benzothiophene structure have been described, of which 14 and 16 are examples. Although 14 and 16 have similar activity by the estrogen receptor such as 17-hydroxy-β-estradiol, little is known about vascular effects. Uchiumi et al., In tl. J. Cancer, 47, 80-85 (1991). We show that 14 and 16 can antagonize the vascular contractile responses induced by the calcium agonist BayK 8644, documenting the antagonist activity of calcium channels. In this respect, 16 was more potent than 14, a conclusion consistent with its ability to inhibit contraction for the calcium agonist BayK 8644 and its marked inhibitory effect on the contractile responses to norepinephrine, serotonin and U46619.

The antagonist activity of calcium channels 14 and 16 is probably unrelated to their ability to bind to the estrogen receptor, for several reasons. First, both compounds had similar affinity in the estrogen receptor (Uchiumi et al., 1991) and even 16 was considered more potent as a calcium channel antagonist than 14. Second, the ability to inhibit contractile responses occurred acutely in these studies, suggesting that a nuclear event was not required. Thus, the present studies establish an acute effect in vitro of 14 and 16 to inhibit calcium channels in vascular tissue, using the rat aorta by demonstrating the inhibition of BayK-induced contraction 8644, an effect independent of the endothelium. The benzothiophene derivative 16 which binds with affinity similar to the estrogen receptor that 17-β-estradiol is a more potent antagonist of calcium channels in vascular tissue than estrogen. In addition, these comparative studies provide additional data to support the argument that blocking calcium channels by 14 and 16 is unrelated to the ability of these agents to bind to the estrogen receptor. If the inhibition of calcium channels contributes to the clinical effects of estrogen, then 14 and 16 may have some important advantages over estrogen as replacement therapy in postmenopausal women.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

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

Claims (15)

1. The use of a compound for the preparation of a medicament for antagonizing or blocking calcium channels in vascular tissue, comprising administering to a warm-blooded animal in need thereof, a pharmaceutically acceptable amount of a compound having the fdrmula (I) wherein R1 and R3 are independently hydrogen, alkyl of 1 to 4 carbon atoms, -CO- (alkyl of 1 to 6 carbon atoms), -CH2Ar, or -COAr, wherein Ar is phenyl or substituted phenyl; R2 is selected from the group consisting of pyrrolidino, hexamethyleneimino, and piperidino; or a pharmaceutically acceptable salt thereof.

2. The use according to claim 1, wherein in said compound, R1 and R3 are independently hydrogen, alkyl of 1 to 4 carbon atoms, -CO- (alkyl of 1 to 6 carbon atoms), or benzyl; and R2 is piperidino or pyrrolidino.

3. The use according to claim 2, wherein R1 and R3 are independently hydrogen or alkyl of 1 to 4 carbon atoms, and R2 is piperidino or pyrrolidino.

4. The use according to claim 3, wherein R1 and R3 are hydrogen and R2 is piperidino or pyrrolidino.

5. The use according to claim 4, wherein R2 is piperidino.

6. The use according to claim 5, wherein the pharmaceutically acceptable salt is the hydrochloride salt.

7. The use according to claim 4, wherein R is pyrrolidino.

8. The use according to claim 7, wherein the pharmaceutically acceptable salt is the hydrochloride salt.

9. The use according to claim 1, wherein the compound is for the preparation of a medicament for the treatment of cardiac disorders.

10. The use according to claim 9, wherein the compound is for the preparation of a medicament for the treatment of cardiac disorders such as angina variant, angina induced by physical exercise, unstable angina, myocardial damage by ischemia-reperfusion, and arrhythmias. .

11. The use according to claim 1, wherein the compound is for the preparation of a medicament for treating cerebral vascular disorders.

12. The use according to claim 11, wherein the compound is for the preparation of a medicament for the treatment of cerebral vascular disorders such as cerebral vasospasm due to arterial rupture, shock, and migraine headache.

13. The use according to claim 1, wherein the compound is for the preparation of a medicament for the treatment of renal disorders.

14. The use according to claim 1, wherein the compound is for the preparation of a medicament for the treatment of gastrointestinal disorders.

15. The use according to claim 1, wherein the compound is for the preparation of a medicament for the treatment of hypertension.