USE OF ALPHA-1C SPECIFIC COMPOUNDS TO TREAT BENIGN PROSTATIC HYPERPLASIA

Abstract: A method of treating benign prostatic hyperplasia in a subject which comprises administering to the subject a therapeutically effective amount of a compound which binds to a human alpha 1C adrenergic receptor with a binding affinity greater than ten-fold higher than the binding affinity with which the compound binds to a human alpha 1A adrenergic receptor, a human alpha 1B adrenergic receptor and a human histamine H1 receptor, and, binds to a human alpha 2 adrenergic receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to such alpha 1C adrenergic receptor. Compounds meeting these criteria are provided.

Description: USE OF ALPHA-1C SPECIFIC COMPOUNDS TO TREAT BENIGN PROSTATIC HYPERPLASIA Background of the invention Benign Prostatic Hyperplasia (BPH) , also called Benign Prostatic Hypertrophy, is a progressive condition which is characterized by a nodular enlargement of prostatic tissue resulting in obstruction of the urethra. This results in increased frequency of urination, nocturia, a poor urine stream and hesitancy or delay in starting the urine flow. Chronic consequences of BPH can include hypertrophy of bladder smooth muscle, a decompensated bladder and an increased incidence of urinary tract infection. The specific biochemical, histological and pharmacological properties of the prostate adenoma leading to the bladder outlet obstruction are not yet known. However, the development of BPH is considered to be an inescapable phenomenon for the aging male population. BPH is observed in approximately 70% of males over the age of 70. Currently, in the United States, the method of choice for treating BPH is surgery (Lepor, H. Urol. Clinics North Amer.. 17, 651 (1990)). Over 400,000 prostatectomies are performed annually (data from 1986) . A medicinal alternative to surgery is clearly very desirable. The limitations of surgery for treating BPH include the morbidity rate of an operative procedure in elderly men, persistence or recurrence of obstructive and irritative symptoms, as well as the significant cost of surgery. α-Adrenergic receptors are specific neuroreceptor proteins located in the peripheral and central nervous systems on tissues throughout the body. These receptors are important switches for controlling many physiological functions and, thus, represent important targets for drug development. In fact, many α- adrenergic drugs have been developed over the past 40 years. Examples include clonidine, phenoxybenzamine and prazosin (treatment of hypertension) , naphazoline (nasal decongestant) , and apraclonidine (treating glaucoma) . α-Adrenergic drugs can be broken down into two distinct classes: agonists (clonidine and naphazoline are agonists) , which mimic the receptor activation properties of the endogenous neurotransmitter norepinephrine, and antagonists (phenoxybenzamine and prazosin are antagonists) , which act to block the effects of norepinephrine. Many of these drugs are effective but also produce unwanted side effects (for example, clonidine produces dry mouth and sedation in addition to its antihypertensive effects) . During the past 15 years a more precise understanding of α-adrenergic receptors and their drugs has evolved through increased scientific scrutiny. Prior to 1977, only one α-adrenergic receptor was known to exist. Between 1977 and 1988, it was accepted by the scientific community that at least two α-adrenergic receptors—α, and α 2 —existed in the central and peripheral nervous systems. Since 1988, new techniques in molecular biology have led to the identification of at least six α-adrenergic receptors which exist throughout the central and peripheral nervous systems: α 1A , α 1B , α 1c , α 2A , α 2B and α 2c (Bylund, D.B., FASEB J. , 6, 832 (1992)). It is not known precisely which physiological responses in the body are controlled by each of these receptors. Ih addition, many α-adrenergic drugs that were developed before 1992 are not selective for any particular α-adrenergic receptor. Many of these drugs produce untoward side effects which may be attributed to their poor α-adrenergic receptor selectivity. Since the mid 1970's, nonselective α-antagonists have been prescribed to treat BPH. In 1976, M. Caine, et al. (Brit. J. Urol. , 48, 255 (1976)), reported that the nonselective α-antagonist phenoxybenzamine was useful in relieving the symptoms of BPH. This drug may produce its effects by interacting with α-receptors located on the prostate. However, this drug also produces significant side effects which severely limit its use in treating patients on a chronic basis. More recently, the α-adrenergic antagonists prazosin and terazosin have also been found to be useful for treating BPH. However, these drugs also produce untoward side effects. The most recently approved drug Proscar ™ (Merck) prescribed for BPH is not an α-adrenergic antagonist, but rather acts by blocking 5- α-reductase. While Proscar is able to relieve symptoms, it is effective in only 30% of all patients, and requires a period of up to 6 months to show results. From binding studies using cloned rat α 1A , hamster α 18 , and bovine α 1c receptors, and functional studies of antagonism in vitro using human prostrate, I. Marshall, et al., concluded that the receptor mediating contraction of the human prostrate is of the α 1c subtype (Marshall, I., et al., Brit. Pharmacol. Soc. , (1992)). Furthermore, using cloned human receptors the binding characteristics of the known BPH drugs to various receptor subtypes have been determined, as described more fully hereinafter. Based upon such binding information and additional data, it has been observed that the side effects which occur with the drugs prazosin and terazosin may be due to their poor selectivity for specific α-adrenergic receptors. In contrast, indoramin is a drug which is slightly selective for the human α 1c receptor relative to the other human α-adrenergic receptors, but it also interacts at human histamine HI receptors. This compound produces untoward side effects which may be attributed to its activity at such H 1 receptors. It would be desirable to provide methods and compounds which allow the treatment of BPH but which avoid the production of side effects observed for all currently used medications. From the binding information described hereinafter, it has unexpectedly been discovered that compounds which are specific for an α 1c adrenergic receptor with a binding affinity greater than ten-fold higher than the binding affinity with which the compounds bind to an α 1A adrenergic receptor, a human α 1B adrenergic receptor, and a human histamine H 1 receptor, and (b) bind to an α 2 adrenergic receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compounds bind to such α 1c adrenergic receptor are effective for the treatment of BPH. Furthermore, we have characterized several antagonists selective for the α 1c adrenergic receptor using a rat orthostatic hypotension model to ascertain the vascular effects of drugs which may be indicative of their ability to produce dizziness in patients, and observed that while nonselective alpha 1 antagonists produce significant effects on orthostatic hypotension, selective alpha lc antagonists do not produce significant effects. Brief Description of the Drawings A more complete understanding of the invention and many of its advantages will become apparent by reference to the detailed description which follows when considered in conjunction with the accompanying drawings, wherein: Figure 1 illustrates compounds which are potent antagonists of the cloned human α 1c receptor. Figure 2A illustrates the correlation of inhibition constants (pK { ) for a series of α 1 antagonists at the cloned human α 1A receptors with efficiency of blocking contraction of human prostate tissue (pA 2 ) . Figure 2B illustrates the correlation of inhibition constants (pK { ) for a series of α 1 antagonists at the cloned human α 1B receptors with efficiency of blocking contraction of human prostate tissue (pA 2 ) . Figure 2C illustrates the correlation of inhibition constants (pK,-) for a series of α, antagonists at the cloned human α 1c receptors with efficiency of blocking contraction of human prostate tissue (pA 2 ) . SUBSTITUTE SHEET Summary of the Invention The present invention provides a method of treating benign prostatic hyperplasia in a subject which comprises administering to the subject a therapeutically effective amount of a compound which (a) binds to a human α lc adrenergic receptor with a binding affinity greater than ten-fold higher than the binding affinity with which the compound binds to a human α 1A adrenergic receptor, a human α 1B adrenergic receptor, and a human histamine H, receptor, and (b) binds to a human α 2 adrenergic receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to such α 1c adrenergic receptor. The present invention also provides a method of inhibiting contraction of prostate tissue which comprises contacting the prostate tissue with an effective contraction-inhibiting amount of a compound which (a) binds to a human α 1c adrenergic receptor with a binding affinity greater than ten-fold higher than the binding affinity with which the compound binds to a human α 1A adrenergic receptor, a human α 1B adrenergic receptor, and a human histamine H 1 receptor, and (b) binds to a human α 2 adrenergic receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to such α 1c adrenergic receptor. Detailed Description of the Invention The present invention provides a method of treating benign prostatic hyperplasia in a subject which comprises administering to the subject a therapeutically effective amount of a compound which (a) binds to a human α 1c adrenergic receptor with a binding affinity greater than ten-fold higher than the binding affinity with which the compound binds to a human α 1A adrenergic receptor, a human α 1B adrenergic receptor, and a human histamine H 1 receptor, and (b) binds to a human α 2 adrenergic receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to such α 1c adrenergic receptor. Desirably, the compound used to practice the method of the invention additionally binds to a calcium channel with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the α 1c adrenergic receptor. Alternatively or incrementally, the compound used to practice the method of the invention also binds to a dopamine D 2 receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the α 1c adrenergic receptor. Alternatively or incrementally, the compound used to practice the method of the invention additionally binds to a dopamine D 2 receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the α 1c adrenergic receptor. Alternatively or incrementally, the compound used to practice the method of the invention additionally binds to any serotonin receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the α 1c adrenergic receptor. Alternatively or incrementally, the compound used to practice the method of the invention also binds to a human dopamine D 3 receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the α 1c adrenergic receptor. Alternatively or incrementally, the compound used to practice the method of the invention also binds to a human dopamine D 4 with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the α 1c adrenergic receptor. Alternatively or incrementally, the compound used to practice the method of the invention also binds to a human dopamine D 5 receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the α 1c adrenergic receptor. Alternatively or incrementally, the compound used to practice the method of the invention also does not cause orthostatic fall in blood pressure at a dosage effective to alleviate benign prostatic hyperplasia. Alternatively or incrementally, the compound used to practice the method of the invention also does not cause orthostatic fall in blood pressure in rats at a dosage 10 ug/kg. A number of compounds have been identified or synthesized which are useful in the practice of the invention. For example, the compound has the structure: In another example, the compound has the structure: In still another example, the compound has the structure: In an additional example, the compound has the structure: Included within the scope of the method of treating BPH in accord with the invention are the use of both R and S enantiomers of the compounds described which possess stereogenic centers, as well as the use of pharmaceutically acceptable salts and complexes thereof. The invention also provides a method of inhibiting contraction of prostate tissue which comprises contacting the prostate tissue with an effective contraction-inhibiting amount of a compound which (a) binds to a human α 1c adrenergic receptor with a binding affinity greater than ten-fold higher than the binding affinity with which the compound binds to a human α 1A adrenergic receptor, a human α 1B adrenergic receptor, and a human histamine H, receptor, and (b) binds to a human α 2 adrenergic receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to such α 1c adrenergic receptor. The activity of compounds at the different human receptors was determined in vitro using cultured cell lines that selectively express the receptor of interest. These cell lines were prepared by transfecting the cloned cDNA or cloned genomic DNA or constructs containing both genomic DNA and cDNA encoding the human α-adrenergic, serotonin, histamine, and dopamine receptors as further described in detail in Example 9 hereinbelow. In connection with this invention, a number of cloned human receptors discussed herein, either as plasmids or as stably transfected cell lines, have been made pursuant to, and in satisfaction of, the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purpose of Patent Procedure, and are made with the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852. Specifically, these deposits have been accorded ATCC Accession Numbers as follows: The data shown in the accompanying Tables 1 and 2 indicate that the α 1c -specific receptor antagonists which satisfy the criteria as defined herein have significant efficacy in the inhibition of contraction of human prostate tissue. This in vitro property is recognized in the art as correlating with efficacy in treating benign prostatic hyperplasia in vivo. The present invention therefore provides a method of treating benign prostatic hyperplasia, which comprises administering a quantity of any of the α 1c receptor antagonists defined as herein in a quantity effective against BPH. The drug may be administered to a patient afflicted with benign prostatic hyperplasia by any conventional route of administration, including, but not limited to, intravenous, intramuscular, oral, subcutaneous, intratumoral, intradermal, and par¬ enteral. The quantity effective against BPH is between 0.001 mg and 10.0 mg per kg of subject body weight. The method of treating BPH disclosed in the present invention may also be carried out using a pharmaceutical composition comprising any of the α 1c receptor antagonists as defined herein and a pharmaceutically acceptable carrier. The composition may contain between 0.05 mg and 500 mg of an α 1c receptor antagonist, and may be constituted into any form suitable for the mode of administration selected. Compositions suitable for oral administration include solid forms, such as pills, capsules, granules, tablets, and powders, and liquid forms, such as solutions, syrups, elixers, and suspensions. Forms useful for parenteral administration include sterile solutions, emulsions, and suspensions. The drug may otherwise be prepared as a sterile solid composition which may be dissolved or suspended at the time of administration using sterile water, saline, or other appropriate sterile injectable medium. Carriers are intended to include necessary and inert binders, suspending agents, lubricants, flavorants, sweeteners, preservatives, dyes, and coatings. Optimal dosages to be administered may be readily determined by those skilled in the art, and will vary with the particular α 1c receptor antagonist in use, the strength of the preparation, the mode of administration, and the advancement of the disease condition. Additional factors depending on the particular patient being treated will result in a need to adjust dosages, including patient age, weight, diet, and time of administration. The following Experimental Details are set forth to aid in an understanding of the invention, and are not intended, and should not be construed, to limit in any way the invention set forth in the claims which follow thereafter. Experimental Details. Prazosin, 5-methylurapidil, and S-niguldipine were obtained from Research Biochemicals, Inc. A30360 (4- fluoro-4-(8-fluoro-l,3,4,5-tetrahydro-2H-pyrido[4,3- b]indol-2-yl)butyrophenone hydrochloride) was obtained from Aldrich Chemical Co. Other compounds were prepared according to the examples which follow. Example l Synthesis of Terazosin Hydrochloride N-(2-Furoyl)piperazine This compound and its preparation has been described in Great Britain Patents 1,390,014 and 1,390,015. Piperazine hexahydrate (194 g, 1 mole) was dissolved in 250 ml H 2 0. The solution was acidified to pH 4.5 with 6 N HC1. Furoyl chloride (130.5 g, 1 mole, Aldrich) was added along with 10% NaOH solution at such a rate that the pH was maintained at 4.5. After 1 hour, the solution was made basic (pH = 8.5) with NaOH solution. The reaction mixture was continuously extracted with chloroform for 36 hours. The CHC1 3 extract was dried over MgS0 4 , and filtered. Distillation gave 108.2 g product (60%), b.p. 132° - 138° C/0.6 mm Hg, m.p. 69° - 70 °C. N-(Tetrahydro-2-furoyl) iperazine The furoylpiperazine of Example 1 was converted to the hydrobromide salt (m.p. 173" - 175° C) . This salt (39.0 g) in 250 ml methyl alcohol and 9.0 g Raney nickel was hydrogenated at 3 at . After uptake of H 2 ceased, the catalyst was filtered, the solvent concentrated, and the residue crystallized from isopropyl alcohol to give 35.2 g. tetrahydrofuroylpiperazine HBr, m.p. 152° - 156 °C. This was suspended in 20 ml H 2 0. Then 10.5 g 50%, NaOH solution was added slowly followed by 2.0 g solid Na 2 C0 3 . This was extracted with 4 x 100 ml portions of warm CHC1 3 . The CHC1 3 extractions were distilled to give 22.5 g tetrahydrofurolylpiperazine, b.p. 120° - 125°C/0.2 mm Hg. 2 [4-(Tetrahydro-2-furoyl)piperazinyl]-4-araino-6,7- dimethoxyquinazoline hydrochloride To 7.00 g 2-chloro-4-amino-6,7-dimethoxyquinazoline (Lancaster Synthesis) in 50 ml methoxyethanol was added 10.8 g, tetrahydrofurolylpiperazine, and the mixture refluxed 3 hours. The clear solution was concentrated and an aqueous solution of potassium bicarbonate was added. The resultant solid that formed was filtered and washed with water. It was then added to methanol and the resulting suspension was acidified with a solution of hydrogen chloride in isopropyl alcohol. The resulting solution was concentrated and the residue crystallized from isopropyl alcohol giving 8.12 g. of product, m.p. 278° - 279°C. Example 2 Preparation of Indoramin 4-Benzamido-l-[2-(3-indolyl)ethylpyridiniu Bromide A solution of 4-benzamidopyridine (1.98 g) and 3-(2- bromoethyl) indole (2.24 g) in EtOH (15 ml) was refluxed for 2 hours, and the crystallized product (3.13 g, mp 264 - 266°C) was collected by filtration from the hot reaction mixture. Recrystallization gave the hydrate. 3-[2-4-Benzamidopiperid-l-yl)ethyl]indole (Indoramin) 4-Benzamido-l-[2-(3-indolyl)ethyl]pyridinium bromide (3.0g) in 91% EtOH (300 ml) containing Et 3 N (0.8 g) was hydrogenated in the presence of freshly prepared W-7 Raney Ni catalyst (ca. 3 g) at 28.12 kg/cm 2 and 50° for 4 hours. After filtering off the catalyst, the filtrate was evaporated and the residue was shaken with CHC1 3 and 2 N NaOH. The resulting insoluble material (1.61 g, mp 203 - 206°C) was collected and dried. Recrystallization from EtOH gave the product (1.34 g) , as colorless needles. Example 3 Preparation of 1- ( 3 -benzoy1propy1 ) - - benzamidopiperidine A mixture of 4-chlorobutyrophenone (447 mg, 2.45 mmol) , 4-benzamidopiperidine (500 mg, 2.45 mmol) and K 2 C0 3 (338 mg, 2.45 mmol) was heated up in boiling water bath for 1 hour. The reaction mixture was portioned between water and CHCl 3 . The organic layer was separated and dried over Na 2 S0 4 . After filtration and removal of solvent, the residue was purified by chromatography (Si0 2 , MeOH:CHCl 3 , 5:95). Recrystallization from AcOEt/hexane gave a white powder (78 mg, 8.2%) . mp 143- 144°C; 1 H NMR (CD 3 OD, 400MHz) δ 1.65 (dq, J^.lδ Hz, J 2 =11.9 Hz, 2H) , 1.90-2.00 (m, 4H) , 2.18 (t, J=11.9 Hz, 2H) , 2.48 (m, 2H) , 3.00-3.10 (m, 4H) , 3.88 (m, IH) , 7.40-8.00 (m, 10H) ; Mass spectrum (M+l) + at m/z 351. Example 4 Preparation of l-[3-(4-chlorobenzoyl)propyl]-4- benzamidopiperidine A mixture of 3-(4-chlorobenzol)propyl bromide (640 mg, 2.45 mmol), 4-benzamidopiperidine (500 mg, 2.45 mmol) and K 2 C0 3 (1.01 g, 7.34 mmol) in 50 ml of acetone was heated up to refluxing condition for 48 hours. The solid was removed by filtration. Concentration of filtrate In vacuo gave a yellowish solid, which was purified by chromatography (Si0 2 , MeOH:CHCl 3 , 5:95) . 320 mg (33.9%) of white powder was obtained E NMR (CDC1 3 , 300 mHz) S 1.46 (dq, J^l.O Hz, J 2 =8.4 Hz, 2H) , 1.90-2.10 (m, 4H) , 2.16 (m, 2H) , 2.43 (t, J=6.9 Hz, 2H) , 2.80-2.90 (m, 2H) , 2.97 (t, J=6.9 Hz, 2H) , 3.97 (m, IH) , 5.92 (d, J=7.8 Hz, IH, N-H) , 7.40-8.00 (m, 9H) ; Product was converted to HC1 salt and recrystallized with MeOH/Et 2 0, mp 243-244°C; Calcd for C 22 H 25 C1N 2 0 2 -HC1-H 2 0: C 60.15, H 6.37, N 6.37; Found: C 60.18, H 6.34, N6.29. Example 5 Preparation of SKF-104856 1-[ (4-Chlorophenyl)thio}-2-propanone Chloroacetone (32.3 g, 0.347 mol) was added to a mixture of 4-chlorothiophenol (50 g, 0.347 mmol) and sodium hydroxide (14 g, 0.347 mol) in water (400 ml) and the mixture was stirred at 25"C for 1 hour. The mixture was extracted with ethyl ether and the organic phase was washed with water, dried with magnesium sulfate and concentrated to give 69 g (99%) of l-[(4- chlorophenyl)thio]-2-propanone. 5-Chloro-3-methylbenzo(b)thiophene l-[ (4-Chlorophenyl)thio)-2-propanone (50 g, 0.25 mol) was added to polyphosphoric acid (300 g) and the mixture was stirred as the temperature was gradually raised to 120"C as an exotherm started. The mixture was stirred at 130°C for 1 hour, diluted with water, extracted with ethyl ether and the organic phase was dried and concentrated. The residue was stirred in methanol (200 ml) , filtered and the filtrate concentrated to give 17.5 g (40%) of 5-chloro-3- methylbenzo(b)thiophene: bp 120°C (0.6 mm Hg) . Ethyl5-chloro-3-methylbenzo (b) thiophene-2-carboxylate n-Butyllithium in hexane (2.6 M, 2.3 ml) was added to a solution of 5-chloro-3-methylbenzo(b)thiophene (1,0 g, 6 mmol) in ethyl ether (20 ml) stirred at 0°C under argon. The mixture was stirred for 30 minutes and transferred slowly under argon pressure to a stirred solution of ethyl chloroformate (0.63 g, 6 mmol) in ethyl ether (20 ml) . The mixture was stirred at 0°C for 30 minutes and at 25°C for 1.5 hours. The mixture was treated with water and the organic phase was dried, concentrated and triturated with hexane to give 1.0 g (67%) of ethyl 5-chloro-3-methylbenzo(b)thiophene-2- carboxylate: mp 92.5 - 94 °C. Ethyl 3-bromomethyl-5-chlorobenzo(b)thiophene-2- carboxylate A mixture of ethyl 5-chloro-3-methylbenzo(b)thiophene- 2-carboxylate (9.0 g, 0.035 mol), N-bromosuccinimide (6.53 g, 0.037 mol) and benzoyl peroxide (130 mg) in carbon tetrachloride (150 ml) was refluxed and illuminated with sunlamp for 2 hours. The resulting suspension was cooled, filtered and the filter cake was triturated with methanol to give 9.9 g, (85%) of the methanol-insoluble ethyl 3-bromomethyl-5- chlorobenzo(b)thiophene-2-carboxylate: mp 148-150°C. Ethyl 5-Chloro-3- [N- ( 2 , 2-dimethoxyethyl ) -N-methyl ( am- inomethyl) ] enzol (b) thiophene-2 -carboxylate A mixture of ethyl 3 -bromomethy 1-5- chlorobenzo(b) thiophene-2 -carboxylate (11 g, 0.033 mol), methylaminoacetaldehyde dimethyl acetal (4.76 g, 0.04 mol) and potassium carbonate (11.4 g, 0.8 mol) in dry acetone (200 ml) was stirred for 48 hours, filtered and the filtrate concentrated to give 11.8 g, (96%) of ethyl 5-chloro-3- (N-2 , 2-dimethoxyethyl) -N- methyl (a inomethyl) benzol (b) thiophene-2 -carboxy late. Ethyl 7-chloro-3,4-dihydro-4-methylthieno[ ,3,2-ef]- [3]benzazepine-2-carboxylate Ethyl 5-chloro-3-[N-(2.2-dimethoxyethyl)-N- methyl (a inomethyl) ]benzo[b_,t.hiophene-2-carboxylate (3.0 g, 8.1 mmol) was added in portions to trifluoromethanesulfonic acid (10 ml) stirred at 0°C under argon. The mixture was stirred at 25°C for 45 minutes and diluted with water. The mixture was basified with aqueous sodium hydroxide and extracted with ethyl ether to give ethyl 7-chloro-3,4-dihydro-4- methylthieno-[4,3,2-ef] [3]benzazepine-2-carboxylate. Ethyl 7-chloro-3,4,5,6-tetrahydro-4-methylthieno[4,3,2- ef] [3]benzazepine-2-carboxylate Diborane in tetrahydrofuaran (1 M, 40 ml) was added to a solution of ethyl 7-chloro-3,4-dihydro-4- methylthieno[4,3,2-ef] [3]benzazepine-2-carboxylate (2.8 g) in tetrahydrofuran (30 ml) stirred at 0°C. The mixture was refluxed for 3 hours and stirred at 25°C for 18 hours, cooled, treated with methanol (50 ml) , refluxed for 18 hours and concentrated. The residue was triturated with ethyl ether-hexane (3:1) to give 1.6 g (84%) of ethyl 7-chloro-3,4,5,6-tetrahydro-4- methylthieno[4,3,2-ef] [3]benzazepine-2-carboxylate:mp 138-140 °C. The free base was treated with hydrogen chloride to give ethyl 7-chloro-3,4,5,6-tetrahydro-4- methylthieno[4,3,2-ef] [3]benzazepine-2-carboxylate hydrochloride: mp 240°C. 7-Chloro-3,4,5,6-tetrahy ro-4-methylthieno[4,3,2- ef] [3]benzazepine-2-methanol A solution of ethyl 7-chloro-3,4,5,6-tetrahydro-4- methylthieno[4.3.2-ef] [3]benzazepine-2-carboxylate (4.0 g, 12.9 mmol), in ethyl ether (48 ml) was treated with lithium aluminum hydride (0.53 g, 14 mmol). The mixture was stirred for 1.5 hours, cooled and treated carefully with water (2.0 ml), 10% sodium hydroxide (1.0 ml) and water (2.0 ml) . The resulting mixture was filtered and the solvent evaporated to give 1.9 g (57%) of 7-chloro-3 ,4,5, 6-tetrahydro-4-methylthieno [4,3,2- ef ] [3]benzazepine-2-methanol: mr 184-185°C. 7 -Chloro- 3 ,4,5, 6-tetrahydro-4-methylthieno-4 ,3,2- ef ] [3]benzazepine-2-carboxaldehyde A solution of 7-chloro-3 , 4 , 5, 6-tetrahydro-4- methylthieno[4,3,2-ef ] [ 3 ]benzazepine-2 -methanol (1.6 g, 6 mmol) in dichloromethane (150 ml) was stirred under argon with activated manganese dioxide (8.3 g) for 2 hours. The mixture was filtered through Celite ™ and the filtrate was dried with magnesium sulfate and concentrated to give a 63% yield of 7-chloro-3, 4 , 5, 6- tetrahydro-4-methylthieno[4 , 3 , 2-ef [ [ 3 ] benzazepine-2- carboxaldehyde . 7-Chloro-2-ethenyl-3 , 4 , 5 , 6-tetrahdyro-4- methylthieno[4,3,2-ef ] [3]benzazepine (SKF-104856) Sodium hydride (60 % dispersion in mineral oil. 3.8 mmol) was added to a stirred solution of methyl triphenylphosphonium bromide (1.35 g, 3.8 mmol) in dry tetrahydrofuran (30 ml) and stirred for 15 minutes. The mixture was treated with a solution of 7- chloro-3,4,5,6-tetrahydro-4-methylthieno[4, 3, 2-ef] [3]- benzazepine-2-carboxaldehyde, prepared as in Example 3, (0.5 g, 1.9 mmol) in dimethyl forma ide (4 ml), stirred at 25 "C for 16 hours, quenched with ice and extracted with ethyl acetate. The organic phase was washed, dried and concentrated and the residue was chromatographed on silica gel eluted with a gradient of methylene chloride to methanol-methylene chloride (3.5:96.5). The product was treated with hydrogen chloride to give 0.2 g (35%) of 7-chloro-2-ethenyl- 3 , 4 , 5 , 6-tetrahydro-4-methylthieno[4 , 3 , 2 - ef ] [3]benzazepine hydrochloride: mp 234-236 °C. Example 6 2-Hydroxymethyl-l,2,3, -tetrahydronaphthalene A solution of 1,2,34-tetrahydro-2-naphthoic acid (2.50 g, 14.2 mmol) in 100 ml THF was treated with LiAlH 4 (681 mg, 17.04 mmol) and the reaction mixture was heated at reflux for 5 hours. The suspension was cooled to O'C and quenched by addition of solid Na 2 SO A Bl0H 2 O. The mixture was stirred at room temperature for 4 hours. The solid was removed by filtration. Concentration of filtrate in vacuo gave a yellowish oil (2.28 g, 98.8%); H NMR (CDCl 3 , 300 MHz) 6 1.43 (m, IH) , 2.00 (m, 2H) 2.51 (dd, J.,=16.5 Hz, J 2 =10.8 Hz, IH) , 2.85 (m, 3H) , 3.65 (dd, J^δ.3 Hz, J 2 =1.2 Hz, 2H) , 7.09 (s, 4H) . 2-Bromomethyl-l,2,3,4-tetrahydronaphthalene A solution of 2-hydroxymethy1-1 , 2 , 3 , 4- tetrahydronaphthalene (2.28 g, 14.0 mmol) in 100 ml of CH 2 C1 2 was treated with PBr 3 (1.28 g, 4.73 mmol) at 0°C. The mixture was stirred at room temperature for 72 hours then poured onto 100 g of ice. The organic layer was isolated, washed with 10% K 2 C0 4 aqueous solution, H 2 0, sat'd brine, and then dried over Na 2 S0 4 . After filtration and removal of solvent, the residue was purified by chromatography (Si0 2 , EtOAc:hexane, 1:10) to give a colorless oil (1.33 g, 41.6%); 1 H NMR (CDC1 3 , 300 MHz) δ 1.55 (m, IH) , 2.11 (m, IH) , 2.11 (m, 2H) , 2.58 (dd, J^lβ.2 Hz, J 2 =10.2 Hz, IH) , 2.80-3.10 (m, 3H) , 3.45 (d, J=6.3 Hz, 2H) , 7.10 (m, 4H) . 2-[(4-Methoxyphenethyl)aminomethyl]-l,2,3,4- tetrahydronaphthalene (Compound 11) A solution of 2 -bromomethy1-1 , 2 , 3 , 4 - tetrahydronaphthalene (1.33 g, 5.91 mmol) and 4- methoxyphenethylamine (1.79 g, 11.8 mmol) in 50 ml of EtOH was refluxed for 48 hours. After removal of EtOH in vacuo, the residue was dissolved in 100 ml of CHC1 3 , washed with 10% K 2 C0 3 , H 2 0, sat'd brine, and then dried over Na 2 S0 4 . Filtration followed by evaporation of solvent gave a yellow oil, which was purified by chromatography (Si0 2 , MeOH:CHCl 3 , 5:95) to a give a yellowish oil (1.03 g, 58.9%). The product was converted to HCl salt, crystallization with MeOH/Et 2 0 gave a white powder. mp 274-275°C; Calcd for C 20 H 25 NO.HC1: C 72.37, H 7.91, N 4.22; Found C 72.40, H 7.76, N 4.13. Example 7 4 ,4-Diphenylpiperidine hydrochloride A mixture of 4-piperidone monohydrate hydrochloride (15.0 g, 97.6 mmol, 1.00 equiv, Aldrich) and A1C1 3 (130 g, 976 mmol, 10.0 equiv) in anhydrous benzene (600 mL) was stirred at reflux for 4 hours. Ice (300 g) and water (50 mL) were added, the mixture was filtered, and the solid was washed with toluene and dried to afford 19.2 g (72%) of off-white solid, which was pure by 1 H NMR. Recrystallization from ethanol gave the analytically pure sample: m.p. 300-301 °C; 1 H NMR (300 MHz, CD 3 0D) δ 2.65 (m, 4 H) , 3.18 (m, 4 H) , 7.18 (m, 2 H) , 7.30 (m, 8 H) ; Anal. Calcd. for C 17 H 19 N-HC1: C, 74.57; H. 7.36; N, 5.12. Found: C, 74.32; H, 7.34; N, 5.02. The free base was generated by addition of the above salt to dilute aqueous sodium hydroxide and extraction with CH 2 C1 2 . The organic phase was dried over MgS0 4 and concentrated to give a light brown solid: IR (neat) 2942.8, 1494.5, 1445.9 cm "1 ; CIMS (NH 3 ) m/e 238 (M + 1) + . 3- (4, -Diphenylpiperidin-l-yl)propionitrile To a suspension of 4, 4-diphenylpiperidine hydrochloride (195 mg, 0.712 mmol, 1.0 equiv) in ETOh (1.5 mL) was added triethylamine (0.25 mL, 1.83 mmol, 2.6 equiv) followed by acrylonitrile (0.13 mL, 2.01 mmol, 2.8 equiv) . The resulting solution was stirred at room temperature under argon for 15 minutes and then concentrated. Water was added, and the mixture was extracted three times with EtOAc. The combined organic extracts were dried over MgS0 4 and concentrated to give 170 mg (87%) of tan solid, which was used for the next reaction without purification. m.p. 95-96°C; 1 H NMR (300 MHz, CDC1 3 ) δ 2.37 (m, 2H) , 2,46 (m, 4H) , 2.52 (m, 6H) , 7.12 (m, 2H) , 7.23 (m, 8H) ; 13 C NMR (75 MHz, CDC1 3 ) δ 16.65, 36.71. 45.08, 50.78, 54.13, 119.70, 126.48, 127.78, 129.11, 147.87; IR (neat) 2944.4, 2821.0, 1495.5, 1445.9 cm '1 . 1-(3-Aminopropyl)-4,4-diphenylpiperidine To a stirred solution of 3-(4,4-diphenylpiperidine-l- yl)propionitrile (2.00 g, 6.89 mmol, 1.0 equiv) in anhydrous THF (20 mL) under argon was added a solution of BH 3 in THF (1.0 M, 24.1 mL, 24 mmol, 3.5 equiv) at room temperature. The mixture was refluxed for 4.5 hours and then cooled to room temperature. Aqueous HCl (6 N, 50 mL) was added and stirring was continued for 1 hour. The mixture was basified to pH 9 by addition of 6 N aq. NaOH, extracted 3 times with CH 2 C1 2 , dried over MgS0 4 and concentrated. The residue was purified by flash chromatography (Si0 2 , EtOAc-MeOH, 9:1, followed by EtOAc-MeOH-isopropylamine (60:10:1), followed by EtOAc-MeOH-isopropylamine (40:10:2) to give 1.35 g (66%) of tan solid: m.p. 98-99°C; 1 H NMR (300 MHz, CDC1 3 ) δ 1.64 (tt, J=7.7 Hz, 2H) , 2.33 (br t, J=7.2 Hz, 2H) , 2.50 ( , 8H) , 2.76 (br t, J=6.5 Hz, 2H) , 3.06 (br s, 2H) , 7.13 (m , 2H) , 7.26 (m, 8H) ; 13 C NMR (75 MHz, CDC1,) δ 29.79, 36.80, 41.41, 45.24, 51.25, 57.41, 126.30, 127.77, 128.97, 148.11; IR (neat) 3361.5 cm '1 ; CIMS (NH 3 ) m/e 295 (M + 1) + . Acetoacetic acid N-[3-(4,4-diphenylpiperidin-l- yl)propyl]amide Diketene (0.44 mL, 5.68 mmol, 1.3 equiv, Aldrich) was added at room temperature to a stirred solution of 1- (3-aminopropyl)-4-,4-diphenylpiperidine (1.288 g, 4.37 mmol, 1.0 equiv) in anhydrous toluene (15 mL) under argon, and stirring was continued for 48 hours. The mixture was concentrated to give 1.294 g (78%) of white solid, which was used for the next reaction without purification: 1 H NMR (300 MHz, CDC1 3 ) δ 1.70 (tt, J=6.4, 6.4 Hz, 2H) , 2.23 (s, 3H) , 2.44 (br t, J=6.5 Hz), 2.49-2.67 (m, 8H) , 3.32 (br t, J=5.8 Hz) , 3.36 (s, 2H) , 7.16 (m, 2H) , 7.27 (m, 8H) . 2 ,6-Dimethyl-4-(4-nitrophenyl)-1, -dihydropyridine-3,5- dicarboxylic acid N-[3-(4,4-diphenylpiperidine-l- yl)propyl]amide methyl ester A solution of acetoacetic acid N-[3-(4,4- diphenylpiperidin-l-yl)propyl]amide (365 mg, 0.964 mmol, 1.0 equiv), methyl 3-aminocrotonate (138 mg, 1.20 mmol, 1.2 equiv, Aldrich), and 4-nitrobenzaldehyde (181 mg, 1.20 mmol, 1.2 equiv, Aldrich) in isopropanol was refluxed under argon for 60 hours. The mixture was cooled to room temperature and concentrated, and the residue was diluted with CH 2 C1 2 , washed with water, dried over MgS0 4 , and concentrated. The residue was purified by flash chromatography (Si0 2 , EtOAc, followed by EtOAc-MeOH, 19:1 and 9:1) to give 147.8 mg (25%) of yellow solid: 1 H NMR (300 MHz, CDC1 3 ) δ 1.55 (m, 2H) , 2.14 (Ξ, 3H) , 2.15-2.50 (m, 10H) , 2.32 (s, 3H) , 3.20 (m, IH) , 3.37 (m, IH) , 3.54 (s, 3H) , 5.00 (s, 3H) , 5.48 (br s) , 6.98 (br t, J=4.9 Hz, IH) , 7.14-7.30 (m, 10H) , 7.39 (dm, J=8.7 Hz, 2H) , 8.05 (dm, J=8.7 Hz, 2H) ; 13 C NMR (75 MHz, CDC1 3 ) δ 18.74, 20.64, 25.61, 36.77, 40.20, 42.26, 45.03, 51.16, 51.61, 58.08, 100.65, 109.71, 124.35, 126.46, 127.61, 128.84, 129.06, 135.52, 146.96, 147.10, 154.55, 168.22, 168.70; IR (neat) 1680, 1610, 1515, 1340 cm "1 ; MS (FAB) m/e 609 (M + H) + . 2,6-Dimethyl-4-(4-nitrophenyl)-l,4-dihydropyridine-3,5- dicarboxylic acid N-[3-(4,4-diphenylpiperidin-l-yl)- propyl]amide methyl ester hydrochloride hydrate (Compound 2) To a solution of 2 ,6-dimethyl-4-(4-nitrophenyl)-1,4- dihydro-pyridine-3,5-dicarboxylic acid N-[3-(4,4- diphenylpiperidin-l-yl)propyl]amidemethyl ester (147.8 mg, 0.243 mmol, 1.0 equiv) in EtOH (2 mL) was added a solution of HCl in ether (1.0 M, 0.24 mL, 0.24 mmol, 1.0 equiv). Addition of ethyl acetate (3 mL) followed by heating gave a clear solution. Slow cooling of this solution, followed by filtration gave 91 mg of yellow crystalline solid: m.p. 182-183°C; Anal. Calcd. for C 36 H 40 N 4 O 5 -HCI ' H 2 O: C ' 65 - 20 / H » 6.54 ; N, 8.45. Found: C, 65.30; H, 6.28; N, 8.15. Example 8 3-(4,4-Diphenylpiperid-l-yl)-propanol 4,4-Diphenylpiperidine (40 g) , 3-bromopropanol (24.7 g, Aldrich), powdered potassium carbonate (116.4 g) and approximately 1 g of potassium iodide (in 500 ml of a 1:1 mixture of dioxane and 1-butanol) were heated for about 48 hours under reflux and with vigorous stirring. After cooling, the mixture was filtered, and the filtrate was concentrated. The oily residue was taken up in ethyl acetate, and the solution was filtered again. Concentrating the filtrate to dryness yielded the product in the form of a yellowish, oily residue which slowly solidifies to a wax-like product (yield: 44.8 g) . Hydrochloric acid in ether produced the hydrochloride (m.p.: 226° to 227° C) , which was recrystallized from 2-propanol. Acetoacetic acid 3- (4,4-diphenylpiperidin-l-yl)propyl ester 23.6 g of 3-(4,4-diphenylpiperid-l-yl)-propanol were dissolved in 100 ml of absolute toluene, and 16 ml of a 50% strength solution of diketene in acetone were added with stirring. After standing for several days at room temperature (monitored by thin layer chromatography) , the mixture was concentrated, and the residue was dried under high vacuum. The pale yellow, viscous oil which remains was employed without further purification for the next stage. 2 ,6-Dimethyl-4- (4-nitrophenyl)-1,4-dihydropyridine-3,5- dicarboxy-ylic acid [3- (4,4-diphenylpiperidin-l- yUpropyl] ester methyl ester A solution of methyl 3-aminocrotonate (265 mg, 2.3 mmol, 1.0 equiv), 4-nitrobenzaldehyde (348 mg, 2.3 mmol, 1.0 equiv), and acetoacetic acid 3-[4, 4- diphenylpiperidin-l-yl)propyl] ester (872 mg, 2.3 mmol, 1.0 equiv) in isopropanol was refluxed under argon with stirring for 68 hours. Cooling and removal of solvent gave a residue, which was purified by flash chromatography (Si0 2 , EtOAc-hexane, 1:1 and 1:2, followed by EtOAc) to afford 717 mg (51%) of yellow solid: 1 H NMR (300 MHz, CDCl 3 ) <5 1.73 (m, 2H) , 2.22 (m, 2H) , 2.30-2.51 (m, 8H) , 2.34 (s, 3H) , 2.35 (s, 3H) , 3.63 (s, 3H) , 4.05 (dt, J=2.1, 7.9 Hz, 2H) , 5.06 (s, IH) , 5.73 (br s, IH) , 7.14 (m, 2H) , 7.27 (m, 8H) , 7.42 (dm, J=8.8 Hz, 2H) , 8.06 (dm, J=8.8 Hz, 2H) ; 13 C NMR (75 MHZ, CDC1 3 ) δ 15.30, 19.65, 26.32, 36.11, 39.88, 44.60, 50.60, 51.12, 55.34, 62.66, 102.99, 107.55, 123.39, 125.67, 127.12, 128.33, 128.65, 144.80, 144.93, 146.36, 147.50, 154.78, 166.91, 167.43; IR (neat) 1698.0, 1684.7, 1517.5, 1345.7 cm "1 ; CIMS (NH 3 ) 610 (M + 1) + , 553, 338. 2,6-Dimethyl-4-(4-nitrophenyl)-1,4-dihydropyridine-3,5- dicarboxylic acid [3-(4,4-diphenylpiperidin-l- yDpropyl] ester methyl ester hydrochloride (Compound 8) To a solution of 2,6-dimethyl-4-(4-nitrophenyl)-1,4- dihydro-pyridine-3,5-dicarboxylic acid [3-(4,4- diphenylpiperidine-1-yl)-propyl] ester methyl ester (710 mg, 1.16 mmol, 1.0 equiv) in EtOH (5 L) was added a solution of HCl in ether (1.0 M, 1.5 mL, 1.5 mmol, 1.3 equiv). The solvents were removed and the residue was dissolved in CH 2 C1 2 . This solution was added dropwise to 25 mL of ether to afford, after filtration, 500 mg of yellow crystalline solid: m.p. 152-153°C. Anal. Calcd. for C 36 H 39 N 3 0 6 •HCl: C, 66.92; H, 6.24; N, 6.50. Found: C, 66.70; H, 5.99; N, 6.27 Example 9 Protocol for the Determination of the Potency of α 1 Antagonists The activity of compounds at the different human receptors was determined in vitro using cultured cell lines that selectively express the receptor of interest. These cell lines were prepared by transfecting the cloned cDNA or cloned genomic DNA or constructs containing both genomic DNA and cDNA encoding the human α-adrenergic, serotonin, histamine, and dopamine receptors as follows: α 1A Human Adrenergic Receptor: The entire coding region of αlA (1719 bp) (Sequence I.D. No. 1), including 150 basepairs of 5" untranslated sequence (5' UT) and 300 bp of 3' untranslated sequence (3' UT) , was cloned into the Ba HI and Clal sites of the polylinker-modified eukaryotic expression vector pCEXV-3, called EXJ.HR. The construct involved the ligation of partial overlapping human lymphocyte genomic and hippocampal cDNA clones: 5' sequence were contained on a 1.2 kb Smal-Xhol genomic fragment (the vector-derived BamHI site was used for subcloning instead of the internal insert-derived Smal site) . and 3' sequences were contained on an 1.3 kb Xhol-Clal cDNA fragment (the Clal site was from the vector polylinker) . Stable cell lines were obtained by cotransfection with the plasmid αlA/EXJ (expression vector containing the αlA receptor gene) and the plasmid pGCcos3neo (plasmid containing the a inoglycoside transferase gene) into LM(tk " ) , CHO, and NIH3T3 cells, using calcium phosphate technique. The cells were grown, in a controlled environment (37°C, 5% C0 2 ) , as monolayers in Dulbecco's modified Eagle's Medium (GIBCO, Grand Island, NY) containing 25mM glucose and supplemented with 10% bovine calf serum, 100 units/ml penicillin g, and 100 μg/ml streptomycin sulfate. Stable clones were then selected for resistance to the antibiotic G-418 (1 mg/ml) , and membranes were harvested and assayed for their ability to bind [ 3 H]prazosin as described below (see "Radioligand Binding assays") . α 1B Human Adrenergic Receptor: The entire coding region of αlB (1563 bp) (Sequence I.D. No. 3), including 200 basepairs and 5' untranslated sequence (5' UT) and 600 bp of 3' untranslated sequence (3" UT) , was cloned into the EcoRI site of pCEXV-3 eukaryotic expression vector. The construct involved ligating the full-length containing EcoRI brainstem cDNA fragment from λ ZapII into the expression vector. Stable cell lines were selected as described above. Human α 1c Adrenergic Receptor: The entire coding region of αlC (1401 bp) (Sequence I.D. No. 5), including 400 basepairs of 5' untranslated sequence (5' UT) and 200 bp of 3' untranslated sequence (3' UT) , was cloned into the Kpnl site of the polylinker-modified pCEXV-3- derived eukaryotic expression vector, EXJ.RH. The construct involved ligating three partial overlapping fragments: a 5' 0.6kb Hindi genomic clone, a central 1.8 EcoRI hippocampal cDNA clone, and a 3' 0.6Kb PstI genomic clone. The hippocampal cDNA fragment overlaps with the 5' and 3' genomic clones so that the Hindi and PstI sites at the 5' and 3' ends of the cDNA clone, respectively, were utilized for ligation. This full- length clone was cloned into the Kpnl site of the expression vector, using the 5' and 3' Kpnl sites of the fragment, derived from vector (i.e., pBluescript) and 3'-untranslated sequences, respectively. Stable cell lines were selected as described above. Radioligand Binding Assays: Transfected cells from culture flasks were scraped into 5ml of 5mM Tris-HCl, 5mM EDTA, pH 7.5, and lysed by sonication. The cell lysates were centrifuged at 1000 rpm for 5 min at 4°C, and the supernatant was centrifuged at 30,000 x g for 20 min at 4°C. The pellet was suspended in 50mM Tris- HCl, ImM MgCl 2 , and 0.1% ascorbic acid at pH 7.5. Binding of the αl antagonist [ 3 H]prazosin (0.5 nM, specific activity 76.2 Ci/mmol) to membrane preparations of LM(tk-) cells was done in a final volume of 0.25 ml and incubated at 37°C for 20 min. Nonspecific binding was determined in the presence of 10 μM phentolamine. The reaction was stopped by filtration through GF/B filters using a cell harvester. Inhibition experiments, routinely consisting of 7 concentrations of the tested compounds, were analyzed using a non-linear regression curve-fitting computer program to obtain Ki values. α 2 Human Adrenergic Receptors: To determine the potency of α 1 antagonists at the α 2 receptors, LM(tk-) cell lines stably transfected with the genes encoding the α 2A , α 2B , and α 2c receptors were used. The cell line expressing the α 2A receptor is designated L-α 2A , and was deposited on November 6, 1992 under ATCC Accession No. CRL 11180. The cell line expressing the α 2B receptor is designated L-NGC-α 2B , and was deposited on October 25, 1989 under ATCC Accession No. CRL10275. The cell line expressing the α 2c receptor is designated L-α 2c , and was deposited on November 6, 1992 under ATCC Accession No. CRL-11181. Cell lysates were prepared as described above (see Radioligand Binding Assays) , and suspended in 25mM glycylglycine buffer (pH 7.6 at room temperature) . Equilibrium competition binding assay were performed using [3H]rauwolscine (0.5nM), and nonspecific binding was determined by incubation with lOμM phentolamine. The bound radioligand was separated by filtration through GF/B filters using a cell harvester. Human Histamine H 1 Receptor: The coding sequence of the human histamine H., receptor, homologous to the bovine H, receptor, was obtained from a human hippocampal cDNA library, and was cloned into the eukaryotic expression vector pCEXV-3. The plasmid DNA for the H 1 receptor is designated pcEXV-Hl, and was deposited on November 6, 1992 under ATCC Accession No. 75346. This construct was transfected into COS-7 cells by the DEAE-dextran method. Cells were harvested after 72 hours and lysed by sonication in 5mM Tris-HCl, 5mM EDTA, pH 7.5. The cell lysates were centrifuged at 1000 rpm for 5 min at 4°C, and the supernatant was centrifuged at 30,000 x g for 20 min. at 4°C. The pellet was suspended in 37.8 mM NaHP0 4 , 12.2 mM KH 2 P0 4 , pH 7.5. The binding of the histamine H, antagonist [ 3 H]mepyramine (InM, specific activity: 24.8 Ci/mM) was done in a final volume of 0.25 ml and incubated at room temperature for 60 min. Nonspecific binding was determined in the presence of 10 μM mepyramine. The bound radioligand was separated by filtration through GF/B filters using a cell harvester. Human Histamine H 2 Receptor: The coding sequence of the human H 2 receptor was obtained from a human placenta genomic library, and cloned into the cloning site of PCEXV-3 eukaryotic expression vector. The plasmid DNA for the H 2 receptor is designated pcEXV-H2, and was deposited on November 6, 1992 under ATCC Accession No. 75346. This construct was transfected into COS-7 cells by the DEAE-dextran method. Cells were harvested after 72 hours and lysed by sonication in 5mM Tris-HCl, 5mM EDTA, pH 7.5. The cell lysates were centrifuged at 1000 rpm for 5 min at 4°C, and the supernatant was centrifuged at 30,000 x g for 20 min at 4 °C. The pellet was suspended in 37.8 mM NaHP0 4 , 12.2 mM K2P0 4 , pH 7.5. The binding of the histamine H 2 antagonist [ 3 H]tiotidine (5nM, specific activity: 70 Ci/mM) was done in a final volume of 0.25 ml and incubated at room temperature for 60 min. Nonspecific binding was determined in the presence of 10 μM histamine. The bound radioligand was separated by filtration through GF/B filters using a cell harvester. Human Serotonin Receptors: 5HT 10β , 5HT 1W , 5HT 1E , 5HT 1F Receptors: The cell lysates of LM(tk-) clonal cell line stably transfected with the genes encoding each of these 5HT receptor-subtypes were prepared as described above. The cell line for the 5HT 1Dα receptor, designated as Ltk-8-30-84, was deposited on April 17, 1990, and accorded ATCC Accession No. CRL 10421. The cell for the 5HT 1W receptor, designated as Ltk-11, was deposited on April 17, 1990, and accorded ATCC Accession No. CRL 10422. The cell line for the 5HT 1E receptor, designated 5 HT 1E - 7, was deposited on November 6, 1991, and accorded ATCC Accession No. CRL 10913. The cell line for the 5HT 1F receptor, designated L-5-HT 1F , was deposited on December 27, 1991, and accorded ATCC Accession No. ATCC 10957. These preparations were suspended in 50mM Tris-HCl buffer (pH 7.4 at 37°C) containing 10 M MgCl 2 , 0.2 M EDTA, lOμM pargyline, and 0.1% ascorbate. The potency of α 1 antagonists was determined in competition binding assay by incubation for 30 minutes at 37°C in the presence of 5nM [3H]serotonin. Nonspecific binding was determined in the presence of lOμM serotonin. The bound radioligand was separated by filtration through GF/B filters using a cell harvester. Human 5HT 2 Receptors: The coding sequence of the human 5HT 2 receptor was obtained from a human brain cortex cDNA library, and cloned into the cloning site of pCEXV-3 eukaryotic expression vector. This construct was transfected into COS-7 cells by the DEAE-dextran method. Cells were harvested after 72 hours and lysed by sonication in 5mM Tris-HCl, 5mM EDTA, pH 7.5. This cell line was deposited with the ATCC on October 31, 1989, designated as L-NGC-5HT 2 , and was accorded ATCC Accession No. CRL 10287. The cell lysates were centrifuged at 1000 rpm for 5 minutes at 4°C, and the supernatant was centrifuged at 30,000 x g for 20 minutes at 4°C. The pellet was suspended in 50mM Tris- HCl buffer (pH 7.7 at room temperature) containing 10 mM MgS0 4 , 0.5mM EDTA, and 0.1% ascorbate. The potency of alpha-1 antagonists at 5HT2 receptors was determined in equilibrium competition binding assays using [3H]ketanserin (InM) . Nonspecific binding was defined by the addition of lOμM mianserin. The bound radioligand was separated by filtration through GF/B filters using a cell harvester. Human Dopamine D2 Receptors: The potency of α 1 antagonists at the D2 receptor was determined using membrane preparations from COS-7 cells transfected with the gene encoding the human D2 receptor. The coding region for the human D2 receptor was obtained from a human striatum cDNA library, and cloned into the cloning site of PCDNA 1 eukariotic expression vector. The plasmid DNA for the D 2 receptor is designated pcEXV-D2, and was deposited on November 6, 1992 under ATCC Accession No. ATC 75344. This construct was transfected into COS-7 cells by the DEAE-dextran method. Cells were harvested after 72 hours and lysed by sonication in 5mM Tris-HCl, 5mM EDTA, pH 7.5. The cell lysates were centrifuged at 1000 rpm for 5 minutes at 4°C, and the supernatant was centrifuged at 30,000 x g for 20 minutes at 4°C. The pellet was suspended in 50 mM Tris-HCl (pH 7.4) containing ImM EDTA, 5mM KCl, 1.5mM CaCl 2 , 4mM MgCl 2 , and 0.1% ascorbic acid. The cell lysates were incubated with [3H]spiperone (2nM) , using lOμM (+)Butaclamol to determine nonspecific binding. Other Dopamine receptors are prepared by known methods (D 3 : Sokoloff, P. et al., Nature, 347, 146 (1990), and deposited with the European Molecular Biological Laboratory (EMBL) Genbank as X53944; D 4 : Van Tol, H.H.M., et al., Nature, 350, 610 (1991), and deposited with EMBL Genbank as X58497; D 5 : Sunahara, R.K. , et al., Nature, 350, 614 (1991), and deposited with EMBL Genbank as X58454-HU HD 5DR) . Determination of the Activity of α 1 Antagonists at Calcium Channels The potency of α 1 antagonists at calcium channels was determined in competition binding assays of [3H]nitrendipine to membrane fragments of rat cardiac muscle, essentially as described by Glossman and Ferry (Methods in Enzy ology 109:513-550, 1985). Briefly, the tissue was minced and homogenized in 50mM Tris-HCl (pH 7.4) containing O.lmM phenylmethylsulfonyl fluoride. The homogenates were centrifuged at 1000 g for 15 minutes, the resulting supernatant was centrifuged at 45,000 g for 15 minutes. The 45,000 g pellet was suspended in buffer and centrifuged a second time. Aliquots of membrane protein were incubated for 30 minutes at 37°C in the presence of [3H]nitrendipine (InM) , and nonspecific binding was determined in the presence of lOμM nifedipine. The bound radioligand was separated by filtration through GF/B filters using a cell harvester. Example 10 Functional Properties of α 1 Antagonists in the Human Prostate The efficacy of α, adrenergic antagonists for the treatment of benign prostatic hyperplasia (BPH) is related to their ability to elicit relaxation of prostate smooth muscle. An index of this efficacy can be obtained by determining the potency of α 1 antagonists to antagonize the contraction of human prostatic tissue induced by an α 1 agonist "in vitro". Furthermore, by comparing the potency of subtype selective α, antagonists in binding assays using human α 1 receptors with their potency to inhibit agonist- induced smooth muscle contraction, it is possible to determine which of the α 1 adrenergic receptor subtypes is involved in the contraction of prostate smooth muscle. Methods: Prostatic adenomas were obtained at the time of surgery from patients with symptomatic BPH. These were cut into longitudinal strips of 15mm long and 2-4 mm wide, and suspended in 5ml organ baths containing Krebs buffer (pH 7.4). The baths were maintained at 37°C and continuously oxygenated with 5% C0 2 and 95% 0 2 . Isometric tension was measured with a Grass Instrument FT03 force transducer interfaced with a computer. Tissue strips were contracted with varying concentrations of phenylephrine after incubating for 20 minutes in the absence and presence of at least three different concentrations of antagonist. Dose-response curves for phenylephrine were constructed, and the antagonist potency (pA 2 ) was estimated by the dose- ratio method. The concentration of some antagonists in the tissue bath was assessed by measuring the displacement of [3H]prazosin by aliquots of the bath medium, using membrane preparations of the cloned human α 1c receptor. This control was necessary to account for losses of antagonist due to adsorption to the tissue bath and/or metabolism during the time the antagonists were equilibrated with the prostate tissue. Results: Table 1 shows that the pA 2 values measured for a series of α_, antagonists in human prostate tissue correlate closely (r=0.76) with the corresponding pK { values measured in the α lc receptor assays. In contrast, the human prostate pA 2 values correlate poorly with the pK i values measured at the α 1A (r=-0.06) and α 1B (r=-0.24) adrenergic receptors. (See Figure 2A-Figure 2C.) Thus, antagonists which are more potent at blocking the α 1c adrenergic receptor are more effective at blocking the contraction of the human prostate than antagonists which are more potent at the α 1A or α 1B adrenergic receptors. In addition, antagonists which are selective for the α 1c receptor will have a better therapeutic ratio than nonselective α antagonists. With SNAP 5036 (11), the low pA 2 observed in the prostate may be attributed to tissue absorption or metabolism. Table 2 illustrates the cross reactivity of α, antagonists at other receptors such as α^, α 2B , α 2c , histamine H,, H 2 , serotonin 5-HT 1Dβ , 5-H 1w , 5-HT 1E , 5- HT 1F , 5-HT 2 , and dopamine D 2 . Only compounds SNAP 5036, 5041, and 5089 have binding affinities which are greater than ten-fold higher at α 1c receptors than the binding affinities at other receptors. SUBSTITUTE SHEET Table 1. COMPARISON OF THE BINDING POTENCY (pK,) OF ALPHA-1 ANTAGONISTS IN CLONED HUMAN RECEPTORS ANO THEIR PROTENCV (pλ,) TO INHIBIT PROSTATE SMOOTH MUSCLE CONTRACTION I Table 2. CROSS REACTIVITY OF ALPHA-1 ANTAGONISTS AT CLONED HUMAN RECEPTORS <pκ. ) I ω oo I ND=Not Determined Example 11 Functional Properties of α 1 Antagonists on Rat Orthostatic Hypertension We have identified a large series of compounds (well over 150 compounds, data not shown) which exemplify the hereinabove described properties of antagonists highly selective for the α 1c adrenergic receptor. That is, these compounds are highly selective Alpha lc antagonists which have less than 10 fold the affinity at cloned human Alpha la, Alpha lb, Alpha 2a, Alpha 2b, Alpha 2c, Histamine HI, Dopamine D2 and Serotonin receptors. In addition, these compounds have 10 fold lower affinity at calcium channels (data not shown) . We designated five of these highly selective antagonists for the α 1c adrenergic receptor as drugs 21- 25 and used them to further characterize highly selective antagonists for the α lc adrenergic receptor. In addition, a number of these selective alpha lc antagonists are potent at inhibiting the phenylephrine stimulated contraction of human prostate as described in Example 10. This is a well established protocol for evaluation the efficacy of drugs which may be useful for the treatment of BPH. In addition, we have examined a number of selective alpha lc antagonists in an in vivo canine prostate model (Felson, D. , et al., J. Urol.. 141, 1230-1233 (1989))which is a well characterized model for evaluating the efficacy of BPH drugs (data not shown) . In this model, selective alpha lc antagonists increase urethral pressure at doses which do not produce significant decreases in canine blood pressure. In contrast, nonselective alpha 1 antagonists do not have as large a separation between the effects on urethral pressure and the effects on blood pressure. These observations support our premise that a selective alpha lc antagonist will have a better safety profile than a nonselective alpha 1 antagonist. We have further characterized selective alpha lc antagonists in a rat orthostatic hypotension model. This model gives information on the vascular effects of drugs which may be indicative of their ability to produce dizziness in patients (Hieble, J.P., et al.. Cardiovascular Pharmacology. 15, 845 (1990)). Our objective was to characterize the effects of selective alpha lc antagonists on rat orthostatic hypotension and contrast the results with those obtained using nonselective alpha 1 antagonists. Methods Rat Orthostatic Hypotension Model Adult male Sprague-Dawley normotensive rats were anesthetized with sodium pentobarbital (45mg/kg, i.v. ) . The femoral vein and artery of the right hindlimb were cannulated for drug administration and blood pressure monitoring, respectively. Heart rate was determined by a cardiotachometer triggered by the blood pressure pulse. The rats were secured in the supine position to a board that could be tilted 90 degrees. When blood pressure and heart rate had stabilized, the rats were subjected to a 90 degree vertical (head up) tilt for 60 seconds. Changes in blood pressure and heart rate from pre-tilt levels were monitored continuously. The rats were returned to the supine position and blood pressure and heart rate were allowed to stabilize. Either an antagonist selective for the α 1c adrenergic receptor (designated drug 21, 22, 23, 24 or 25), an antagonist nonselective for the α 1c adrenergic receptor (Prazosin or Terazosin) or saline was then administered through venous cannula, either as an i.v. bolus or as an infusion. When blood pressure had stabilized, the rats were subjected to a second tilt and blood pressure and heart rate were recorded as described above. Most saline treated rats typically exhibit a greater ability to return their blood pressure toward pre-tilt levels during the second tilt. Data from the second tilt are used in statistical analysis. Results Table 3 shows that while nonselective alpha 1 antagonists produce significant effects on orthostatic hypotension, selective alpha lc antagonists do not produce significant effects. More specifically, Prazosin and Terazosin consistently cause orthostasis at the lowest dose (lOug/kg) and, in some rats, in a dose-dependent manner. Drug 21 causes orthostasis only at the highest dose (1000 ug/kg) in 2 out of 4 rats, while the other antagonists selective for the α 1c adrenergic receptor caused no orthostasis at the highest dose. Placebo and 22, 23, 24, 25 did not induce orthostasis at any dose. Taken all together, this is a positive result since it is believed that orthostatic hypotension contributes to the dizziness observed clinically with nonselective alpha 1 antagonists. This further supports our premise that a selective alpha lc antagonist will have a better safety profile than a nonselective alpha 1 antagonist. TABLE 3 Summary of Studies on Drug Effects on Orthostasis I I + and - mean positive or negative indings, respective y +, ++ and +++ are relative to doses of the same drug but not compared to other drugs +/- positive findings found in some rats SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Charles Gluchowski, et al. (ii) TITLE OF INVENTION: Use of Alpha-lC Specific Compounds to Treat Benign Prostatic Hyperplasia (iii) NUMBER OF SEQUENCES: 6 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: COOPER & DUNHAM (B) STREET: 30 Rockefeller Plaza (C) CITY: New York (D) STATE: New York (E) COUNTRY: U.S.A. (F) ZIP: 10112 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.24 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: White, John P. (B) REGISTRATION NUMBER: 28,678 (C) REFERENCE/DOCKET NUMBER: 376901 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (212) 977-9550 (B) TELEFAX: (212) 664-0525 (C) TELEX: (212) 422523 COOP Ul (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2140 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N (iv) ANTI-SENSE: N (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 178..1893 (D) OTHER INFORMATION: -44- CCGGGCCλGG CACGTCCGCT CTCGGACAGC CGCTCCGCGT CACAGGAACT TGGGCAGGAC 60 CCGACGGGAC CCGTGCGCGG AGCTGCATCT GGAGCCCCGC GGCTATGCCC TGTGCTCCCC 120 TCCTGCCGGC CGCTCGTTCT GTGCCCCCGG CCCGGCCACC GACGGCCGCG CGTTGAG 177 ATG ACT TTC CGC GAT CTC CTG AGC GTC AGT TTC GAG GGA CCC CGC CCG 225 Met Thr Phe Arg Aβp Leu Leu Ser Val Ser Phe Glu Gly Pro Arg Pro 1 5 10 15 GAC AGC AGC GCA GGG GGC TCC AGC GCG GGC GGC GGC GGG GGC AGC GCG 273 Aβp Ser Ser Ala Gly Gly Ser Ser Ala Gly Gly Gly Gly Gly Ser Ala 20 25 30 GGC GGC GCG GCC CCC TOG GAG GGC CCG GCG GTG GGC GGC GTG CCG GGG 321 Gly Gly Ala Ala Pro Ser Glu Gly Pro Ala Val Gly Gly Val Pro Gly 35 40 45 GGC GCG GGC GGC GGC GGC GGC GTG GTG GGC GCA GGC AGC GGC GAG GAC 369 Gly Ala Gly Gly Gly Gly Gly Val Val Gly Ala Gly Ser Gly Glu Asp SO 55 60 AAC CGG AGC TCC GCG GGG GAG CCG GGG AGC CCG GGC GCG GGC GGC GAC 417 Aan Arg Ser Ser Ala Gly Glu Pro Gly Ser Ala Gly Ala Gly Gly Asp 65 70 75 80 GTG AAT GGC ACG GCG GCC GTC GGG GGA CTG GTG GTG AGC GCG CAG GGC 465 Val Asn Gly Thr Ala Ala Val Gly Gly Leu Val Val Ser Ala Gin Gly 85 90 95 GTG GGC GTG GGC GTC TTC CTG GCA GCC TTC ATC CTT ATG GCC GTG GCA 513 Val Gly Val Gly Val Phe Leu Ala Ala Phe He Leu Met Ala Val Ala 100 105 110 GGT AAC CTG CTT GTC ATC CTC TCA GTG GCC TGC AAC CGC CAC CTG CAG 561 Gly Aβn Leu Leu Val He Leu Ser Val Ala Cys Asn Arg Hie Leu G n 115 120 12S ACC GTC ACC AAC TAT TTC ATC GTG AAC CTG GCC GTG GCC GAC CTG CTC 609 Thr Val Thr λβn Tyr Phe He Val Aβn Leu Ala Val Ala Aβp Leu Leu 130 135 140 CTG AGC GCC ACC GTA CTC CCC TTC TCC GCC ACC ATG GAG GTT CTG GGC 657 Leu Ser Ala Thr Val Leu Pro Phe Ser Ala Thr Met Glu Val Leu Gly 145 150 155 160 TTC TGG GCC TTT GGC CGC GCC TTC TGC GAC GTA TGG GCC GCC GTG GAC 705 Phe Trp Ala Phe Gly Arg Ala Phe Cyβ Aβp Val Trp Ala Ala Val Aβp 165 170 175 GTG CTG TGC TGC ACG GCC TCC ATC CTC AGC CTC TGC ACC ATC TCC GTC 753 Val Leu Cys Cys Thr Ala Ser He Leu Ser Leu Cys Thr He Ser Val 180 185 190 GAC CGG TAC GTG GGC GTG CGC CAC TCA CTC AAG TAC CCA GCC ATC ATG 801 Aβp Arg Tyr Val Gly Val Arg His Ser Leu Lye Tyr Pro Ala He Met 195 200 205 ACC GAG CGC AAG GCG GCC GCC ATC CTG GCC CTG CTC TGG GTC GTA GCC 849 Thr Glu Arg Lye Ala Ala Ala He Leu Ala Leu Leu Trp Val Val Ala 210 215 220 -45- CTC GTC GTG TCC GTA GGG CCC CTG CTC GGC TGG AAG GAG CCC GTG CCC 897 Leu Val Val Ser Val Gly Pro Leu Leu Gly Trp Lyi Glu Pro Val Pro 225 230 235 240 CCT GAC GAG CGC TTC TGC GGT ATC ACC GAG GAG GCG GGC TAC GCT GTC 945 Pro Aβp Glu Arg Phe Cys Gly He Thr Glu Glu Ala Gly Tyr Ala Val 245 250 255 TTC TCC TCC GTG TGC TCC TTC TAC CTG CCC ATG GCG GTC ATC GTG GTC 993 Phe Ser Ser Val Cyβ Ser Phe Tyr Leu Pro Met Ala Val He Val Val 260 265 270 ATG TAC TGC CGC GTC TAC GTG GTC GCG CGC AGC ACC ACG CGC AGC CTC 1041 Met Tyr Cyβ Arg Val Tyr Val Val Ala Arg Ser Thr Thr Arg Ser Leu 275 280 285 GAG GCA GGC GTC AAG CGC GAG CGA GGC AAG GCC TCC GAG CTG GTG CTG 1089 Glu Ala Gly Val Lye Arg Glu Arg Gly Lye Ala Ser Glu Val Val Leu 290 295 300 CGC ATC CAC TGT CGC GGC GCG GCC ACG GGC GCC GAC GGG GCG CAC GGC 1137 Arg He Hie Cyβ Arg Gly Ala Ala Thr Gly Ala Aβp Gly Ala Hie Gly 305 310 315 320 ATG CGC AGC GCC AAG GGC CAC ACC TTC CGC AGC TCC CTC TCC GTC CGC 1185 Met Arg Ser Ala Lys Gly His Thr Phe Arg Ser Ser Leu Ser Val Arg 325 330 335 CTG CTC AAC TTC TCC CGT GAG AAG AAA GCG GCC AAG ACT CTC GCC ATC 1233 Leu Leu Lys Phe Ser Arg Glu Lys Lys Ala Ala Lys Thr Leu Ala He 340 345 350 GTC GTG GGT GTC TTC GTG CTC TGC TGG TTC CCT TTC TTC TTT GTC CTG 1281 Val Val Gly Val Phe Val Leu Cys Trp Phe Pro Phe Phe Phe Val Leu 355 360 365 CCG CTC GGC TCC TTG TTC CCG CAG CTG AAG CCA TCG GAG GGC GTC TTC 1329 Pro Leu Gly Ser Leu Phe Pro Gin Leu Lys Pro Ser Glu Gly Val Phe 370 375 380 AAG GTC ATC TTC TGG CTC GGC TAC TTC AAC AGC TGC GTC AAC CCG CTC 1377 Lys Val He Phe Trp Leu Gly Tyr Phe Asn Ser Cys Val Aβn Pro Leu 385 390 395 400 ATC TAC CCC TGT TCC AGC CGC GAG TTC AAG CGC GCC TTC CTC CGT CTC 1425 He Tyr Pro Cys ser Ser Arg Glu Phe Lys Arg Ala Phe Leu Arg Leu 405 410 415 CTG CGC TCC CAC TGC CGT CGT CGC CGG CGC CGC CGC CCT CTC TGG CGT 1473 Leu Arg Cys Gin Cys Arg Arg Arg Arg Arg Arg Arg Pro Leu Trp Arg 420 425 430 GTC TAC GGC CAC CAC TGG CGG GCC TCC ACC AGC GGC CTG CGC CAG GAC 1521 Val Tyr Gly His Bis Trp Arg Ala Ser Thr Ser Gly Leu Arg Gin Asp 435 440 445 TGC GCC CCG AGT TCG GGC GAC GCG CCC CCC GGA GCG CCG CTG GCC CTC 1569 Cyβ Ala Pro Ser Ser Gly Asp Ala Pro Pro Gly Ala Pro Leu Ala Leu 450 455 460 1 950 -46- ACC GCG CTC CCC GAC CCC GAC CCC GAA CCC CCA GGC ACG CCC GAG ATG 1617 Thr Ala Leu Pro Aβp Pro Aβp Pro Glu Pro Pro Gly Thr Pro Glu Met 465 470 475 480 CAG GCT CCC GTC GCC AGC CGT CGA AAG CCA CCC AGC GCC TTC CGC GAG 1665 Gin Ala Pro Val Ala Ser Arg Arg Lys Pro Pro Ser Ala Phe Arg Glu 485 490 495 TGG AGG CTG CTG GGG CCG TTC CGG AGA CCC ACG ACC CAG CTG CGC GCC 1713, Trp Arg Leu Leu Gly Pro Phe Arg Arg Pro Thr Thr Gin Leu Arg Ala 500 505 510 AAA GTC TCC AGC CTG TCG CAC AAG ATC CGC GCC GGG GGC GCG CAG CGC 1761 Lye Val Ser Ser Leu Ser Hie Lye He Arg Ala Gly Gly Ala Gin Arg 515 520 525 GCA GAG GCA GCG TGC GCC CAG CGC TCA GAG GTG GAG GCT GTG TCC CTA 1809 Ala Glu Ala Ala Cyβ Ala Gin Arg Ser Glu Val Glu Ala Val Ser Leu 530 535 540 GGC GTC CCA CAC GAG GTC GCC GAG GGC GCC ACC TGC CAG GCC TAC GAA 1857 Gly Val Pro Hie Glu Val Ala Glu Gly Ala Thr Cyβ Gin Ala Tyr Glu 545 550 555 560 TTG GCC GAC TAC AGC AAC CTA CGG GAG ACC GAT ATT TAAGGACCCC 1903 Leu Ala Aβp Tyr Ser Aβn Leu Arg Glu Thr Asp He 565 570 AGAGCTAGGC CGCGGAGTGT GCTGGGCTTG GGGGTAAGGG GGACCAGAGA GGCGGGCTGC 1963 TGTTCTAAGA GCCCCCGTGC AAATCGGAGA CCCGGAAACT GATCAGGGCA GCTGCTCTGT 2023 GACATCCCTG AGGAACTGGG CAGAGCTTGA GGCTGGAGCC CTTGAAAGGT GAAAAGTAGT 2083 GGGGCCCCCT GCTGGACTCA GGTGCCCAGA ACTCTTTTCT TAGAAGGGAG AGGCTGC 2140 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 572 amino acidβ (B) TYPE: amino acid (0) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Met Thr Phe Arg Aβp Leu Leu Ser Val Ser Phe Glu Gly Pro Arg Pro 1 5 10 15 Aβp Ser Ser Ala Gly Gly Ser Ser Ala Gly Gly Gly Gly Gly Ser Ala 20 25 30 Gly Gly Ala Ala Pro Ser Glu Gly Pro Ala Val Gly Gly Val Pro Gly 35 40 45 Gly Ala Gly Gly Gly Gly Gly Val Val Gly Ala Gly Ser Gly Glu Asp 50 55 60 -47- Aβn Arg Ser Ser Ala Gly Glu Pro Gly Ser Ala Gly Ala Gly Gly Aβp 65 70 75 80 Val Aβn Gly Thr Ala Ala Val Gly Gly Leu Val Val Ser Ala Gin Gly 85 90 95 Val Gly Val Gly Val Phe Leu Ala Ala Phe He Leu Met Ala Val Ala 100 105 110 Gly λβn Leu Leu Val He Leu Ser Val Ala Cyβ Asn Arg Hie Leu Gin 115 120 125 Thr Val Thr λβn Tyr Phe He Val Aβn Leu Ala Val Ala Asp Leu Leu 130 135 140 Leu Ser Ala Thr Val Leu Pro Phe Ser Ala Thr Met Glu Val Leu Gly 145 150 155 160 Phe Trp Ala Phe Gly Arg Ala Phe Cyβ Aap Val Trp Ala Ala Val Aβp 165 170 175 Val Leu Cys Cyβ Thr Ala Ser He Leu Ser Leu Cyβ Thr He Ser Val 180 185 190 λβp λrg Tyr Val Gly Val Arg Hie Ser Leu Lye Tyr Pro Ala He Met 19S 200 205 Thr Glu λrg Lys λla λla Ala He Leu Ala Leu Leu Trp Val Val Ala 210 215 220 Leu Val Val Ser Val Gly Pro Leu Leu Gly Trp Lys Glu Pro Val Pro 225 230 235 240 Pro λβp Glu λrg Phe Cyβ Gly He Thr Glu Glu λla Gly Tyr λla Val 245 250 255 Phe Ser Ser Val Cyβ Ser Phe Tyr Leu Pro Met λla Val He Val Val 260 265 270 Met Tyr Cyβ λrg Val Tyr Val Val λla Arg Ser Thr Thr λrg Ser Leu 275 280 285 Glu λla Gly Val Lys λrg Glu λrg Gly Lys λla Ser Glu Val Val Leu 290 295 300 λrg He His Cys λrg Gly λla λla Thr Gly λla λβp Gly λla Hie Gly 30S 310 315 320 Met λrg Ser λla Lye Gly Hie Thr Phe Arg Ser Ser Leu Ser Val λrg 325 330 335 Leu Leu Lys Phe Ser λrg Glu Lys Lye λla λla Lye Thr Leu λla He 340 345 350 Val Val Gly Val Phe Val Leu Cyβ Trp Phe Pro Phe P.he Phe Val Leu 355 360 365 Pro Leu Gly Ser Leu Phe Pro Gin Leu Lye Pro Ser Glu Gly Val Phe 370 375 380 Lye Val He Phe Trp Leu Gly Tyr Phe Aβn Ser Cyβ Val Aβn Pro Leu 385 390 395 400 PCI7US93/10950 -48- He Tyr Pro Cyβ Ser Ser Arg Glu Phe Lye Arg Ala Phe Leu λrg Leu 405 410 415 Leu λrg Cyβ Gin Cyβ λrg λrg λrg λrg λrg Arg λrg Pro Leu Trp λrg 420 425 430 Val Tyr Gly Hie Hie Trp λrg λla Ser Thr Ser Gly Leu λrg Gin Aβp 435 440 445 Cyβ λla Pro Ser Ser Gly λβp λla Pro Pro Gly λla Pro Leu λla Leu 450 455 460 Thr Ala Leu Pro Aβp Pro Aβp Pro Glu Pro Pro Gly Thr Pro Glu Met 465 470 475 480 Gin λla Pro Val λla Ser λrg λrg Lye Pro Pro Ser Ala Phe Arg Glu 485 490 495 Trp Arg Leu Leu Gly Pro Phe Arg Arg Pro Thr Thr Gin Leu Arg λla 500 505 510 Lye Val Ser Ser Leu Ser Hie Lye He Arg Ala Gly Gly λla Gin λrg 515 520 525 λla Glu λla λla Cyβ λla Gin λrg Ser Glu Val Glu λla Val Ser Leu 530 535 540 Gly Val Pro Hie Glu Val λla Glu Gly λla Thr Cyβ Gin λla Tyr Glu 545 550 555 560 Leu λla λβp Tyr Ser λβn Leu λrg Glu Thr λβo He 565 570 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (λ) LENGTH: 1738 base pairβ (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY) unknown (ii) MOLECULE TYPE: DNλ (genomic) (iii) HYPOTHETICAL: H ( iv) AN I -SENSE: N ( ix) FEATURE* (λ) NAME/KEY: CDS (B) LOCATION: 124. . 1683 (D) OTHER INFORMATION : (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3 : GCCAGGλGGG CGCCTCTGGG λλGAAGλCCA CGGGGGAAGC AAAGTTTCAG GGCAGCTGAG 60 GAGCCTTCGC CGCAGCCCTT CCGλGCCCλλ TCλTCCCCCλ GGCTλTGGAG GGCGGACTCT 120 AAG ATG AAT CCC GAC CTC GAC ACC GGC CAC AAC ACA TCλ GCλ CCT GCC 168 Met λβn Pro λβp Leu λβp Thr Gly Hie Aβn Thr Ser Ala Pro λla 1 5 10 15 CλC TGG GGλ GAG TTC Aλλ λλT GCC λλC TTC λCT GGC CCC λλC CAG ACC 216 Hie Trp Gly Glu Leu Lye λβn λla λen Phe Thr Gly Pro λβn Gin Thr 20 25 30 TCG AGC AAC TCC ACA CTG CCC CAG CTG GAC ATC ACC AGG GCC ATC TCT 264 Ser Ser Aβn Ser Thr Leu Pro Gin Leu Aβp He Thr Arg λla He Ser 35 40 45 GTG GGC CTG GTG CTG GGC GCC TTC ATC CTC TTT GCC ATC GTG GGC AAC 312 Val Gly Leu Val Leu Gly Ala Phe He Leu Phe Ala He Val Gly Asn 50 55 60 ATC CTA GTC ATC TTG TCT GTG GCC TGC AAC CGG CAC CTG CGG ACG CCC 360 He Leu Val He Leu Ser Val λla Cyβ λβn λrg Hie Leu λrg Thr Pro 65 70 75 λCC λλC TλC TTC λTT GTC λλC CTG GCC ATG GCC GAC CTG CTG TTG AGC 408 Thr Aβn Tyr Phe He Val Aβn Leu Ala Met λla λsp Leu Leu Leu Ser 80 85 90 95 TTC λCC GTC CTG CCC TTC TCλ GCG GCC CTλ GλG GTG CTC GGC TλC TGG 456 Phe Thr Val Leu Pro Phe Ser λla λla Leu Glu Val Leu Gly Tyr Trp 100 105 110 GTG CTG GGG CGG λTC TTC TGT GλC λTC TGG GCλ GCC GTG GλT GTC CTG 504 Val Leu Gly λrg He Phe Cys λβp He Trp λla λla Val λβp Val Leu 115 120 125 TGC TGC λCλ GCG TCC λTT CTG λGC CTG TGC GCC λTC TCC λTC GλT CGC 552 Cyβ Cyβ Thr λla Ser He Leu Ser Leu Cyβ λla He Ser He λβp λrg 130 135 140 TλC λTC GGG GTC CGC TλC TCT CTG CλG TλT CCC λCG CTG GTC λCC CGC 600 Tyr He Gly Val λrg Tyr Ser Leu Gin Tyr Pro Thr Leu Val Thr λrg 145 150 155 λGC AAG GCC ATC TTG GCG CTG CTC AGT GTC TGG GTC TTG TCC ACC GTC 648 Arg Lye λla He Leu λla Leu Leu Ser Val Trp Val Leu Ser Thr Val 160 165 170 175 λTC TCC λTC GGC CCT CTC CTT GGG TGG λλG GλG CCG GCλ CCC λλC GλT 696 He Ser He Gly Pro Leu Leu Gly Trp Lye Glu Pro λla Pro Aβn λβp 180 185 190 GλC λλG GλG TCC GGC GTC λCC Gλλ Gλλ CCC TTC TλT GCC CTC TTC TCC 744 λβp Lye Glu Cyβ Gly Val Thr Glu Glu Pro Phe Tyr λla Leu Phe Ser 195 200 205 TCT CTG GGC TCC TTC TλC λTC CCT CTG GCG GTC λTT CTλ GTC ATG TAC 792 Ser Leu Gly Ser Phe Tyr He Pro Leu λla Val He Leu Val Met Tyr 210 215 220 TGC CGT GTC TAT ATλ GTG GCC λλG AGA ACC ACC AAG AAC CTA GAG GCA 840 Cyβ Arg Val Tyr He Val λla Lye λrg Thr Thr Lye Asn Leu Glu Ala 225 230 235 GGλ GTC λTC λλG GλG λTG TCC λλC TCC AAG GAG CTG ACC CTG AGG ATC 888 Gly Val Met Lye Glu Met Ser Aβn Ser Lye Glu Leu Thr Leu Arg He 240 245 250 255 -50- CAT TCC AAG λλC TTT CλC GλG GAC ACC CTT AGC AGT ACC AAG GCC AλG 936 Hie Ser Lye λen Phe Hie Glu λβp Thr Leu Ser Ser Thr Lye λla Lye 260 265 270 GGC CAC λλC CCC λGC λGT TCC λTλ GCT GTC λλλ CTT TTT AAG TTC TCC 984 Gly Hie λβn Pro λrg Ser Ser He λla Val Lye Leu Phe Lye Phe Ser 275 280 285 AGG GAλ AAG Aλλ GCλ GCT AAG ACG TTG GGC ATT GTG GTC GGT ATG TTC 1032 Arg Glu Lye Lye λla λla Lye Thr Leu Gly He Val Val Gly Met Phe 290 295 300 ATC TTG TGC TGG CTA CCC TTC TTC ATC GCT CTA CCG CTT GGC TCC TTG 1080 He Leu Cyβ Trp Leu Pro Phe Phe He Ala Leu Pro Leu Gly Ser Leu 305 310 315 TTC TCC ACC CTG AAG CCC CCC GAC GCC GTG TTC AAG GTG GTG TTC TGG 1128 Phe Ser Thr Leu Lye Pro Pro Aβp Ala Val Phe Lye Val Val Phe Trp 320 325 330 335 CTG GGC TλC TTC AAC AGC TGC CTC AAC CCC ATC ATC TAC CCA TGC TCC 1176 Leu Gly Tyr Phe Aβn Ser Cyβ Leu Aβn Pro He He Tyr Pro Cyβ Ser 340 345 350 AGC AλG GλG TTC λλG CGC GCT TTC GTG CGC λTC CTC GGG TGC CλG TGC 1224 Ser Lye Glu Phe Lye λrg λla Phe Val λrg He Leu Gly Cyβ Gin Cys 355 360 365 CGC GGC CGC GGC CGC CGC CGλ CGC CGC CGC CGC CGT CGC CTG GGC GGC 1272 λrg Gly λrg Gly λrg λrg λrg λrg Arg Arg Arg Arg Arg Leu Gly Gly 370 375 380 TGC GCC TAC λCC TAC CGG CCG TGG ACG CGC GGC GGC TCG CTG GλG CGC 1320 Cyβ λla Tyr Thr Tyr λrg Pro Trp Thr λrg Gly Gly Ser Leu Glu λrg 385 390 395 TCG CλG TCG CGC λλG GλC TCG CTG GλC GλC λGC GGC AGC TGC CTG AGC 1368 Ser Gin Ser λrg Lys λβp Ser Leu λβp λβp Ser Gly Ser Cyβ Leu Ser 400 405 410 415 GGC λGC CλG CGG λCC CTG CCC TCG GCC TCG CCG AGC CCG GGC TAC CTC 1416 Gly Ser Gin λrg Thr Leu Pro Ser λla Ser Pro Ser Pro Gly Tyr Leu 420 425 430 GGC CGC GGC GCC CCλ CCG CCλ GTC GAG CTG TGC GCC TTC CCC GAG TGG 1464 Gly Arg Gly λla Pro Pro. Pro Val Glu Leu Cyβ λla Phe Pro Glu Trp 435 440 445 λλG GCG CCC GGC GCC CTC CTG λGC CTG CCC GCG CCT GAG CCC CCC GGC 1512 Lys Ala Pro Gly λla Leu Leu Ser Leu Pro λla Pro Glu Pro Pro Gly 450 455 460 CGC CGC GGC CGC CλC GλC TCG GGC CCG CTC TTC ACC TTC AAG CTC CTG 1560 Arg Arg Gly Arg Hie Aβp Ser Gly Pro Leu Phe Thr Phe Lye Leu Leu 465 470 475 ACC GAG CCC GAG AGC CCC GGG ACC GAC GGC GGC GCC AGC AAC GGA GGC 1608 Thr Glu Pro Glu Ser Pro Gly Thr Aβp Gly Gly Ala Ser Asn Gly Gly 480 485 490 495 -51- TGC GλG GCC GCC GCC GλC GTG GCC AAC GGG CAG CCG GGC TTC Aλλ λGC 1656 Cyβ Glu λla λla λla Aβp Val Ala Aβn Gly Gin Pro Gly Phe Lye Ser 500 505 510 AAC ATG CCC CTG GCG CCC GGG CAG TTT TAGGGCCCCC GTGCGCAGCT 1703 λβn Met Pro Leu λla Pro Gly Gin Phe 515 520 TTCTTTCCCT GGGGAGGAAA λCATCGTGGG GGGGA 1738 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 520 amino acidβ (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: Met Aβn Pro Asp Leu Aβp Thr Gly Hie Aβn Thr Ser Ala Pro λla Hie 1 5 10 15 Trp Gly Glu Leu Lye λβn Ala λβn Phe Thr Gly Pro Asn Gin Thr Ser 20 25 30 Ser λβn Ser Thr Leu Pro Gin Leu λβp He Thr λrg λla He Ser Val 35 40 45 Gly Leu Val Leu Gly λla Phe He Leu Phe λla He Val Gly λβn He 50 55 60 Leu Val He Leu Ser Val λla Cyβ λβn λrg Hie Leu λrg Thr Pro Thr 65 70 75 80 λβn Tyr Phe He Val λβn Leu λla Met λla λβp Leu Leu Leu Ser Phe 85 90 95 Thr Val Leu Pro Phe Ser λla λla Leu Glu Val Leu Gly Tyr Trp Val 100 105 110 Leu Gly λrg He Phe Cyβ λβp He Trp λla λla Val λβp Val Leu Cys 115 120 125 Cyβ Thr λla Ser He Leu Ser Leu Cys λla He Ser He λsp λrg Tyr 130 135 140 He Gly Val λrg Tyr Ser Leu Gin Tyr Pro Thr Leu Val Thr λrg λrg 145 150 155 160 Lye λla He Leu λla Leu Leu Ser Val Trp Val Leu Ser Thr Val He 165 170 175 Ser He Gly Pro Leu Leu Gly Trp Lye Glu Pro Ala Pro Aβn Asp Aβp 180 185 190 Lye Glu Cyβ Gly Val Thr Glu Glu Pro Phe Tyr Ala Leu Phe Ser Ser 195 200 205 -52- Leu Gly Ser Phe Tyr He Pro Leu Ala Val He Leu Val Met Tyr Cyβ 210 215 220 λrg Val Tyr He Val λla Lye λrg Thr Thr Lye Aβn Leu Glu Ala Gly 225 230 235 240 Val Met Lye Glu Met Ser Aβn Ser Lye Glu Leu Thr Leu λrg He Hie 245 250 255 Ser Lye Aβn Phe Hie Glu Aβp Thr Leu Ser Ser Thr Lye Ala Lye Gly 260 265 270 Hie λβn Pro λrg Ser Ser He λla Val Lye Leu Phe Lye Phe Ser λrg 275 280 285 Glu Lye Lye λla λla Lye Thr Leu Gly He Val Val Gly Met Phe He 2S0 295 300 Leu Cyβ Trp Leu Pro Phe Phe He λla Leu Pro Leu Gly Ser Leu Phe 305 310 315 320 Ser Thr Leu Lye Pro Pro Aβp Ala Val Phe Lye Val Val Phe Trp Leu 325 330 335 Gly Tyr Phe Aan Ser Cyβ Leu Aβn Pro He He Tyr Pro Cyβ Ser Ser 340 34S 350 Lys Glu Phe Lye λrg λla Phe Val λrg He Leu Gly Cyβ Gin Cyβ λrg 355 360 365 Gly λrg Gly λrg λrg λrg λrg λrg λrg λrg λrg λrg Leu Gly Gly Cys 370 375 380 λla Tyr Thr Tyr λrg Pro Trp Thr λrg Gly Gly Ser Leu Glu λrg Ser 385 390 395 400 Gin Ser λrg Lye λβp Ser Leu λβp λβp Ser Gly Ser Cyβ Leu Ser Gly 405 410 415 Ser Gin Arg Thr Leu Pro Ser λla Ser Pro Ser Pro Gly Tyr Leu Gly 420 425 430 λrg Gly Ala Pro Pro Pro Val Glu Leu Cys Ala Phe Pro Glu Trp Lys 435 440 445 λla Pro Gly λla Leu Leu Ser Leu Pro λla Pro Glu Pro Pro Gly λrg 4S0 455 460 λrg Gly λrg His λβp Ser Gly Pro Leu Phe Thr Phe Lye Leu Leu Thr 465 470 475 480 Glu Pro Glu Ser Pro Gly Thr λsp Gly Gly λla Ser Asn Gly Gly Cye 485 490 495 Glu λla λla λla λβp Val λla λβn Gly Gin Pro Gly Phe Lye Ser λβn 500 505 510 Met Pro Leu λla Pro Gly Gin Phe 515 520 (2) INFORMλTION FOR SEQ ID NO:5: -53- (i) SEQUENCE CHλRACTERISTICS: (λ) LENGTH: 1639 baβe paire (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNλ (genomic) (iii) HYPOTHETICλL: N (iv) λNTI-SENSE: N (ix) FEλTURE: (λ) NAME/KEY: CDS (B) LOCλTION: 126..1523 (D) OTHER INFORMATION: ( i) SEQUENCE DESCRIPTION: SEQ ID NO:5: CCAGCCAλλC CACTGGCλGG CTCCCTCCAG CCGAGACCTT TTATTCCCGG CTCCCGAGCT 60 CCGCCTCCGC GCCAGCCCGG GAGGTGGCCC TGACAGCCGG ACCTCGCCCG GCCCCGGCTG 120 GGACC ATG GTG TTT CTC TCG GGλ λλT GCT TCC GλC λGC TCC λλC TGC 167 Met Val Phe Leu Ser Gly λβn λla Ser λβp Ser Ser λβn Cyβ 1 5 10 λCC Cλλ CCG CCG GCλ CCG GTG λλC λTT TCC λλG GCC λTT CTG CTC GGG 215 Thr Gin Pro Pro λla Pro Val λβn He Ser Lye λla He Leu Leu Gly IS 20 25 30 GTG λTC TTG GGG GGC CTC λTT CTT TTC GGG GTG CTG GGT AAC ATC CTλ 263 Val He Leu Gly Gly Leu He Leu Phe Gly Val Leu Gly λβn He Leu 35 40 45 GTG λTC CTC TCC GTλ GCC TGT CλC CGλ CλC CTG CλC TCλ GTC λCG CλC 311 Val He Leu Ser Val λla Cys His λrg Hie Leu Hie Ser Val Thr His 50 55 60 TλC TλC λTC GTC λλC CTG GCG GTG GCC GλC CTC CTG CTC ACC TCC ACG 359 Tyr Tyr He Val λβn Leu λla Val λla λβp Leu Leu Leu Thr Ser Thr 65 70 75 GTG CTG CCC TTC TCC GCC λTC TTC GAG GTC CTA GGC TAC TGG GCC TTC 407 Val Leu Pro Phe Ser λla He Phe Glu Val Leu Gly Tyr Trp λla Phe 80 85 90 GGC λGG GTC TTC TGC λλC λTC TGG GCG GCA GTG GAT GTG CTG TGC TGC 455 Gly λrg Val Phe Cyβ λβn He Trp Ala Ala Val Asp Val Leu Cyβ Cyβ 95 100 105 110 ACC GCG TCC ATC ATG GGC CTC TGC ATC ATC TCC ATC GAC CGC TAC ATC 503 Thr λla Ser He Met Gly Leu Cyβ He lie Ser He Asp Arg Tyr He 115 120 125 GGC GTG AGC TAC CCC CTG CGC TAC CCA ACC ATC GTC ACC CAG AGG AGG 551 Gly Val Ser Tyr Pro Leu Arg Tyr Pro Thr He Val Thr Gin Arg Arg 130 135 140 -54- GGT CTC ATG GCT CTG CTC TGC GTC TGG GCA CTC TCC CTG GTC ATA TCC 599 Gly Leu Met λla Leu Leu Cyβ Val Trp Ala Leu Ser Leu Val He Ser 145 150 155 λTT GGλ CCC CTC TTC GGC TGG AGG CAG CCG GCC CCC GAG GAC GλG ACC 647 He Gly Pro Leu Phe Gly Trp Arg Gin Pro λla Pro Glu λβp Glu Thr 160 165 170 ATC TGC CAG λTC λλC GAG GAG CCG GGC TAC GTG CTC TTC TCA CCG CTG 695 He Cyβ Gin He Aβn Glu Glu Pro Gly Tyr Val Leu Phe Ser Ala Leu 175 180 185 190 GGC TCC TTC TAC CTG CCT CTG GCC λTC ATC CTG GTC ATG TAC TGC CGC 743 Gly Ser Phe Tyr Leu Pro Leu Ala He He Leu Val Met Tyr Cyβ Arg 195 200 205 GTC TAC GTG GTG GCC AAG AGG GAG AGC CGG GGC CTC AAG TCT GGC CTC 791 Val Tyr Val Val Ala Lye Arg Glu Ser λrg Gly Leu Lye Ser Gly Leu 210 215 220 AλG λCC GλC AAG TCG GAC TCG GAG Cλλ GTG λCG CTC CGC λTC CλT CGG 839 Lye Thr λβp Lye Ser Asp Ser Glu Gin Val Thr Leu Arg He Hie Arg 225 230 235 Aλλ AAC GCC CCG GCλ GGλ GGC λGC GGG ATG GCC AGC GCC AλG λCC λλG 887 Lys λβn λla Pro λla Gly Gly Ser Gly Met λla Ser λla Lye Thr Lye 240 245 250 λCG CλC TTC TCλ GTG λGG CTC CTC λλG TTC TCC CGG GλG λλG λAλ GCC 935 Thr Hie Phe Ser Val λrg Leu Leu Lye Phe Ser λrg Glu Lye Lye λla 255 260 * 265 270 GCC λλλ λCG CTG GGC λTC GTG GTC GGC TGC TTC GTC CTC TGC TGG CTG 983 λla Lye Thr Leu Gly He Val Val Gly Cyβ Phe Val Leu Cyβ Trp Leu 275 280 285 CCT TTT TTC TTλ GTC λTG CCC ATT GGG TCT TTC TTC CCT GλT TTC λλG 1031 Pro Phe Phe Leu Val Met Pro He Gly Ser Phe Phe Pro λβp Phe Lye 290 295 300 CCC TCT Gλλ λCλ GTT TTT λλλ λTλ GTλ TTT TGG CTC GGλ TλT CTλ λλC 1079 Pro Ser Glu Thr Val Phe Lye He Val Phe Trp Leu Gly Tyr Leu λβn 305 310 315 AGC TGC ATC AλC CCC λTC λTλ TλC CCλ TGC TCC λGC Cλλ GλG TTC λλλ 1127 Ser Cys He λβn Pro He He Tyr Pro Cyβ Ser Ser Gin Glu Phe Lye 320 325 330 AAG GCC TTT CλG λλT GTC TTG λGλ λTC CAG TGT CTC TGC AGλ AAG CAG 1175 Lyβ λla Phe Gin λan Val Leu λrg He Gin Cyβ Leu Cyβ λrg Lye Gin 33S 340 345 350 TCT TCC λλλ CλT GCC CTG GGC TλC λCC CTG CAC CCG CCC AGC CAG GCC 1223 Ser Ser Lyβ Hie λla Leu Gly Tyr Thr Leu Hie Pro Pro Ser Gin Ala 355 360 365 GTG GAλ GGG Cλλ CλC AAG GAC ATG GTG CGC ATC CCC GTG GGA TCA AGλ 1271 Val Glu Gly Gin Hie Lyβ λsp Met Val Arg He Pro Val Gly Ser Arg 370 375 380 -55- GAG ACC TTC TλC λGG λTC TCC AAG ACG GAT GGC GTT TGT GAA TGG Aλλ 1319 Glu Thr Phe Tyr λrg He Ser Lyβ Thr Aap Gly Val Cyβ Glu Trp Lyβ 385 390 395 TTT TTC TCT TCC ATG CCC CGT GGA TCT GCC AGG ATT ACA GTG TCC Aλλ 1367 Phe Phe Ser Ser Met Pro Arg Gly Ser Ala λrg He Thr Val Ser Lyβ 400 405 410 GAC CAλ TCC TCC TGT λCC λCλ GCC CGG GTG λGλ AGT AAA AGC TTT TTG 1415 Asp Gin Ser Ser Cyβ Thr Thr Ala Arg Val Arg Ser Lyβ Ser Phe Leu 415 420 425 430 CAG GTC TGC TGC TGT GTA GGG CCC TCA ACC CCC AGC CTT GAC AAG AAC 1463 Gin Val Cyβ Cyβ Cyβ Val Gly Pro Ser Thr Pro Ser Leu Aβp Lyβ Aβn 435 440 445 CAT CAA GTT CCA ACC ATT AAG GTC CAC ACC ATC TCC CTC AGT GAG AAC 1511 Hie Gin Val Pro Thr He Lyβ Val Hie Thr He Ser Leu Ser Glu Aβn 450 455 460 GGG GAG GAA GTC TAGGλCλGGλ λλGλTGCAGA GGλλλGGGGλ λTATCTTAGG 1563 Gly Glu Glu Val 465 TλCCλTλCCC TGGλGTTCTλ GλGGλTTCCT CGλCλλGCTT λTTCCGλTCC λGλCλTGλTλ 1623 GλTλCλTTGλ TGλGTT 1639 (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 466 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: : Met Val Phe Leu Ser Gly λβn λla Ser Asp Ser Ser Aβn Cyβ Thr Gin 1 5 10 15 Pro Pro λla Pro Val λβn He Ser Lyβ λla He Leu Leu Gly Val He 20 25 30 Leu Gly Gly Leu He Leu Phe Gly Val Leu Gly λβn He Leu Val He 35 40 45 Leu Ser Val λla Cyβ Hie λrg His Leu His Ser Val Thr Hie Tyr Tyr 50 55 60 He Val λβn Leu λla Val λla λβp Leu Leu Leu Thr Ser Thr Val Leu 65 70 75 80 Pro Phe Ser λla He Phe Glu Val Leu Gly Tyr Trp λla Phe Gly λrg 85 90 95 Val Phe Cyβ Aβn He Trp Ala Ala Val Aβp Val Leu Cyβ Cyβ Thr Ala 100 105 110 -56- Ser He Met Gly Leu Cyβ He He Ser He Aβp Arg Tyr He Gly Val 115 120 125 Ser Tyr Pro Leu Arg Tyr Pro Thr He Val Thr Gin Arg Arg Gly Leu 130 135 140 Met Ala Leu Leu Cyβ Val Trp Ala Leu Ser Leu Val lie Ser He Gly 145 150 155 160 Pro Leu Phe Gly Trp Arg Gin Pro Ala Pro Glu Aβp Glu Thr He Cyβ 165 170 175 Gin He Asn Glu Glu Pro Gly Tyr Val Leu Phe Ser Ala Leu Gly Ser 180 185 190 Phe Tyr Leu Pro Leu Ala He He Leu Val Met Tyr Cyβ Arg Val Tyr 195 200 205 Val Val λla Lya Arg Glu Ser Arg Gly Leu Lyβ Ser Gly Leu Lyβ Thr 210 215 220 Asp Lys Ser Asp Ser Glu Gin Val Thr Leu Arg He Hie Arg Lys Asn 225 230 235 240 Ala Pro λla Gly Gly Ser Gly Met λla Ser λla Lyβ Thr Lyβ Thr Hie 245 250 255 Phe Ser Val λrg Leu Leu Lyβ Phe Ser λrg Glu Lyβ Lyβ λla λla Lye 260 265 270 Thr Leu Gly He Val Val Gly Cyβ Phe Val Leu Cyβ Trp Leu Pro Phe 275 280 285 Phe Leu Val Met Pro He Gly Ser Phe Phe Pro λβp Phe Lyβ Pro Ser 290 295 300 Glu Thr Val Phe Lyβ He Val Phe Trp Leu Gly Tyr Leu λβn Ser Cyβ 305 310 315 320 He λβn Pro He He Tyr Pro Cyβ Ser Ser Gin Glu Phe Lys Lys λla 325 330 335 Phe Gin λβn Val Leu λrg He Gin Cyβ Leu Cyβ λrg Lyβ Gin Ser Ser 340 345 350 Lyβ His λla Leu Gly Tyr.Thr Leu Hie Pro Pro Ser Gin λla Val Glu 355 360 365 Gly Gin Hie Lys λβp Met Val λrg He Pro Val Gly Ser λrg Glu Thr 370 37S 380 Phe Tyr λrg He Ser Lyβ Thr Aβp Gly Val Cyβ Glu Trp Lyβ Phe Phe 385 390 395 400 Ser Ser Met Pro Arg Gly Ser Ala Arg He Thr Val Ser Lyβ Asp Gin 405 410 415 Ser Ser Cyβ Thr Thr Ala Arg Val λrg Ser Lyβ Ser Phe Leu Gin Val 420 425 430 Cyβ Cyβ Cyβ Val Gly Pro Ser Thr Pro Ser Leu Asp Lyβ Asn Hie Gin 435 440 445 Val Pro Thr He Lyβ Val Hie Thr He S. '.eu Ser Glu Aβn Gly Glu 450 455 460 Glu Val 465

Claims: hat is claimed is; 1. A method of treating benign prostatic hyperplasia in a subject which comprises administering to the subject a therapeutically effective amount of a compound which: a. binds to a human 1c adrenergic receptor with a binding affinity greater than ten-fold higher than the binding affinity with which the compound binds to a human α 1A adrenergic receptor, a human α 1B adrenergic receptor, and a human histamine H 1 receptor; and b. binds to a human a. adrenergic receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to such α 1c adrenergic receptor. 2. A method of claim 1, wherein the compound additionally binds to a calcium channel with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the 1c adrenergic receptor. 3. A method of claim 1 or 2, wherein the compound additionally binds to a dopamine D 2 receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the α 1c adrenergic receptor. 4. A method of claim 1, 2, or 3, wherein the compound additionally binds to any serotonin receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the lc adrenergic receptor. 5. A method of claim 1, 2, 3, or 4, wherein the compound additionally binds to a dopamine D 3 , D 4 , or D 5 receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to the 1c adrenergic receptor. 6. A method of claim 1, 2, 3, 4, or 5 wherein the compound additionally does not cause orthostatic fall in blood pressure at dosages effective to alleviate benign prostatic hyperplasia. 7. The method of claim 6 wherein the compound additionally does not cause orthostatic fall in blood pressure in rats at a dosage of 10 microgra s of compound per kilogram of rat. 8. A method of claim 1, wherein the compound has the structure: 0CH-, 9. A method of claim 1, wherein the compound has the structure: 10. A method of claim 1, wherein the compound has the structure: 11. A method of claim 1, wherein the compound has the structure: 12. A method of inhibiting contraction of prostate tissue which comprises contacting the prostate tissue with an effective contraction-inhibiting amount of a compound which: a. binds to a human α 1c adrenergic receptor with a binding affinity greater than ten-fold higher than the binding affinity with which the compound binds to a human α 1A adrenergic receptor, a human α 1B adrenergic receptor, and a human histamine H 1 receptor; and b. binds to a human α 2 adrenergic receptor with a binding affinity which is greater than ten-fold lower than the binding affinity with which the compound binds to such α 1c adrenergic receptor.

Inventors: BRANCHEK THERESA A, CHIU GEORGE, FORRAY CARLOS C, GLUCHOWSKI CHARLES, HARTIG PAUL R, WETZEL JOHN M, BRANCHEK, THERESA, A., CHIU, GEORGE, FORRAY, CARLOS, C., GLUCHOWSKI, CHARLES, HARTIG, PAUL, R., WETZEL, JOHN, M.
Applicants: BRANCHEK THERESA A, CHIU GEORGE, FORRAY CARLOS C, GLUCHOWSKI CHARLES, HARTIG PAUL R, SYNAPTIC PHARMA CORP, WETZEL JOHN M, BRANCHEK, THERESA, A., CHIU, GEORGE, FORRAY, CARLOS, C., GLUCHOWSKI, CHARLES, HARTIG, PAUL, R., SYNAPTIC PHARMACEUTICAL CORPORATION, WETZEL, JOHN, M.
Application year: 1993
Main classification: A61K31
IPC codes: A61K31, A61K45, A61P13, A61P15, A61P43, C07C211, C07C215, C07C217, C07D211, C07D401, C07D405, C07D495