JP2000506527A - Methods for treating or preventing interstitial cystitis - Google Patents

Abstract

(57) Abstract: The present invention is a method for treating or preventing interstitial cystitis or urethral syndrome in a mammal, comprising administering to a mammal in need thereof an effective amount of a formula: Wherein R ¹ and R ² are independently selected from the group consisting of hydrogen, methyl, methoxy, chloro and trifluoromethyl (provided that only one of R ¹ and R ² can be hydrogen ), Y is A compound represented by NR ^a or CH-NR ^b R ^c (where R ^a , R ^b and R ^c are independently selected from the group consisting of hydrogen and C ₁ -C ₆ alkyl)] Or administering a pharmaceutically acceptable salt or solvate thereof.

Description

DETAILED DESCRIPTION OF THE INVENTION                       Methods for treating or preventing interstitial cystitisBackground of the Invention   Tachykinins share a common amidated carboxy-terminal sequence. It is a peptide family. Substance P is first isolated in this family Should have been purified, and its primary sequence determined until the early 1970s. It was not done in.   From 1983 to 1984, a number of groups were A (also known as substance K, neuromedin L, and neurokinin α) And neurokinin B (neuromedin K and neurokinin β) Two new types of tachykinins, also known as The authors reported their isolation (for a review of these findings, see J.E.Maggio, Peptides, 6 (extra issue 3): 237-243 (1985)).   Tachykinins are widely distributed in both the central and peripheral nervous systems, are released from the nerves, In most cases, it depends on the activation of specific receptors expressed on the membrane of the target tissue. Exert various biological effects. Tachykinin is also produced in many non-neural tissues You.   Mammalian tachykinins substance P, neurokinin A and new Lokinin B has three major components, called NK-1, NK-2 and NK-3, respectively. It acts through various receptor subtypes. These receptors are found in various organs Exists.   Above all, Substance P is used for pain sensation, including pain associated with migraine and arthritis. It is thought to be involved in neurotransmission. These peptides are useful for inflammatory bowel disease. It is also involved in gastrointestinal diseases and disorders. Tachykinin is used in various forms described below. It also plays a role in other diseases.   Tachykinins are used to mediate the sensation and transmission of pain or nociception, especially migraine Playing a major role (eg, S.L. Shepheard et al., British Journal of Pharmacology, 108: 11-20 (1993); S.M.Moussaoui et al., European Journal of Pharmacology, 238: 421-424 (1993) and W.S.Lee et al., British Journal of Pharmacology, 112: 920-924 (1994)).   In view of various diseases related to tachykinin excess, tachykinin receptor The development of a gonist will help control these clinical conditions. Earliest The tachykinin receptor antagonists were peptide derivatives. The antago Nist is considered to have limited pharmaceutical utility due to metabolic instability Was.   Recent publications generally state that early tachykinin receptor antagonists Better oral bioavailability and metabolic stability than Las A new class of non-peptidyl tachykinin receptor antagonists with Was posted. Such a new non-peptidyl tachykinin receptor antagonist Examples are US Pat. No. 5,328,927 (issued Jul. 12, 1994) and US Pat. No. 5,360,820 (November 1, 1994), US Patent No. 5,344,830 (issued on September 6, 1994) No. 5,331,089 (issued on July 19, 1994), European Patent Publication No. 591,040A1 (1994 Published on April 6), European Patent Publication No. WO 94/01402 (published on January 20, 1994), Publication No. WO94 / 04494 (published March 3, 1994) and Patent Cooperation Treaty Publication No. WO93 / 011609 (published on January 21, 1993).   Interstitial cystitis is a chronic debilitating inflammatory disorder of the bladder. The disease is typically about Most common in women between the ages of about 30 and 60 with onset of a medical condition occurring in their 40s Seen in Interstitial cystitis is a bladder associated with morphological and histological changes in the bladder Suprapubic compression and pain with filling, pollakiuria, nocturia, dysuria, emergency and irritating voiding It is characterized by many urological disorders such as pain. The condition does not affect the bladder surface It is thought that the intercellular space (or stroma) in the inner lining of the bladder is thought to be affected, Interstitial cystitis ".   Urethral syndrome is associated with pain of unknown etiology affecting women with various above mentioned conditions Dysuria.   U.S. Pat. No. 5,145,859, issued Sep. 8, 1992, the contents of which are incorporated herein by reference. Contrary to the etiology of interstitial cystitis and urethral syndrome Many compounds have been proposed for treating such conditions, based on the theory. At present, none of these treatments is considered completely effective. Present Currently there are complaints in the affected population about currently available interstitial cystitis treatments There is a need for more effective and safer treatments.Summary of the Invention   The present invention is directed to treating or preventing interstitial cystitis or urethral syndrome in a mammal. For treating a mammal in need thereof with an effective amount of a compound of formula I: [Wherein, R¹And R^TwoAre independently hydrogen, methyl, methoxy, chloro and trif Selected from the group consisting of fluoromethyl (provided that R¹And R^TwoOnly one is water Can be elementary), Y is NR^aOr CH-NR^bR^c(Where R^a, R^bAnd R^cIndependently Hydrogen and C₁-C₆Selected from the group consisting of alkyl) Or a pharmaceutically acceptable salt or solvate thereof. And a method comprising:Detailed description and preferred embodiments   Terms and abbreviations used in this example have their ordinary meanings unless otherwise specified. For example, “° C.” is a degree Celsius, “N” represents a definition or a normality, “Mol” represents mol or mol (s), “mmol” is mmol or mmol (G), gram (g) or gram (s), and kg (kg) Represents gram or kilogram (s), where “L” is liter or liter (multiple) Number), "mL" means milliliter or milliliter (s), “M” represents moles or molarity, “MS” represents mass spectrometry, and “NM” "R" stands for nuclear magnetic resonance analysis.   As used herein, the term "C₁-C₆"Alkyl" is a straight-chain or Represents a branched monovalent saturated aliphatic chain, which includes methyl, ethyl, propyl, Propyl, butyl, isobutyl, t-butyl, pentyl, isopentyl and Includes but is not limited to xyl. The term "C₁-C₆Alkyl " Includes the term "C₁-C_ThreeAlkyl ".   "Halo" represents chloro, fluoro, bromo or iodo.   As used herein, the term "haloformate" refers to an ester of haloformic acid . This compound has the formula: Wherein X is halo and R^dIs C₁-C₆Alkyl]. Preferred c Roformate is bromoformate and chloroformate. Especially preferred Is chloroformate. R^dIs C_Three-C₆An alkyl haloformate is Particularly preferred. Most preferred is isobutyl chloroformate.   The compound produced by the method of the present invention has an asymmetric center. Have this chiral center As a result, the compounds produced in the present invention are racemates, enantiomers As mixtures and individual enantiomers as well as diastereomers and dimers Could occur as an astereomeric mixture. Such asymmetric, individual Methods for producing isomers and mixtures thereof are within the scope of the present invention.   The terms "R" and "S" as used herein are commonly used in organic chemistry. And shows a specific arrangement of the chiral center. The term "R" (rectus (Right)) indicates the superiority of the group when looking at the group with the lowest priority along the bond. Chiral centers with a clockwise relationship (highest to second lowest) Show the arrangement. The term "S" (sinister (left)) has the lowest priority along the join Looking at the base, the counter-clockwise relationship between the base priority (highest to second Shows the configuration of the chiral center having the lowest). The priority of a group is its atomic number (In order of decreasing atomic number). Some lists of priorities and stereochemistry Discussion of the NOMENCLATURE OF ORGANIC COMPOUNDS: PRINCIPLES AND PRACTICE (Edited by J.H. Fletcher et al., 1974), pp. 103-120.   In the present specification, in addition to the (R)-(S) method, the old DL method is also used particularly for amino acids. Used to indicate absolute configuration. In this way the Fischer projection formula is The first carbon is oriented such that it is on top. The prefix "D" is a functional (determining) group Is used to denote the absolute configuration of the isomer to the right of the chiral center carbon atom, "L" is used to denote the absolute configuration of the isomer to the left.   As used herein, the term "treating (treating)" (or "treatment (treatment)" Do) includes alleviating, stopping or reversing the progression, severity or symptoms Invert, prevent, prevent, and control (Restraining) including its generally accepted meaning. this As such, the methods of the present invention include both therapeutic and prophylactic administration.   In particular, Patent Cooperation Treaty Publication WO95 / 14017 (published May 26, 1995) has the formula II : Wherein m and n are independently 0-6, Z is-(CHR^Four)_p− (CHR⁶)_q(Where p is 0 or 1; q is 0 or 1;^FourAnd R⁶Is independently hydrogen and C₁-C_ThreeAlkyl Selected from the group consisting of Y is NR^aOr CH-NR^bR^c(Where R^a, R^bAnd R^cIs independently water Prime and C₁-C₆Selected from the group consisting of alkyl), R¹And R^TwoIs independently hydrogen, halo, C₁-C₆Alkoxy, C₁-C₆Alkyl E, nitro, trifluoroomethyl, or C₁-C₆Is alkyl] A series of tachykinin receptor antagonists or their pharmaceutically acceptable Disclose salts or solvates. These compounds are very active, specific It has been shown to be a Nin receptor antagonist. Particularly preferred compounds Is such that m and n are both 1 and R¹And R^TwoAre independently hydrogen, methoxy, d Toshiki, chloro, fluoro, trifluoromethyl, methyl and ethyl; Z is methylene and Y is 4- when bonded to the heterocyclic group to which it is bonded. (Piperidin-1-yl) piperidin-1-yl, 4- (cyclohexyl) piperazi 1-yl, 4- (phenyl) piperazin-1-yl or 4- (phenyl) pi A compound of formula II that forms peridin-1-yl.   Particularly preferred is the compound (R) -3- (1H-indol-3-yl) -1-. [N- (2-methoxybenzyl) acetylamino] -2- [N- (2- (4-piperidi 1-yl) piperidin-1-yl) acetyl) amino] propane and pharmaceuticals thereof Salts and solvates. Most preferred is compound (R) -3- (1H-Indol-3-yl) -1- [N- (2-methoxybenzyl) acetyla Mino] -2- [N- (2- (4-piperidin-1-yl) piperidin-1-yl) ace Tyl) amino] propane dihydrochloride trihydrate.   The most preferred synthesis method of this compound is shown in the following reaction formula I. Various aspects of this synthesis method The process of the Patent Cooperation Treaty Publication No. WO95 / 14017 (published May 26, 1995) and European Patent No. 693,489 (published Jan. 24, 1996).Reaction Formula I [In the formula, “Tr” represents a trityl group, and “NMM” represents N-methylmorpholine. You. ]Reaction formula I (continued) (R) -2- [N- (2-((4-cyclohexyl) piperazin-1-yl) acetyl ) Amino] -3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) ) Acetylamino] propane  (A) (R) -3- (1H-indol-3-yl) -2- (N-triphenyl Production of [methylamino] propanoic acid [N-trityltryptophan]Tritylation   Chlorotrimethylsilane (70.0 mL, 0.527 mol) in a nitrogen environment D-tryptophan (10 mL) in anhydrous methylene chloride (800 mL) at moderate speed. (0.0 g, 0.490 mol). This mixture is added to 4.25 Stirred continuously for hours. Triethylamine (147.0 mL, 1.055 mol ) And then triphenylmethyl chloride in methylene chloride (400 mL) (147.0 g, 0.552 mol) was added using an addition funnel. mixture Was stirred at room temperature under a nitrogen environment for at least 20 hours. Methanol (500m L) was added to stop the reaction.   The solution is concentrated to almost dryness on a rotary evaporator and the mixture is Redissolved in Tylene and ethyl acetate. Then a 5% citric acid solution (2X) and Aqueous work-up was carried out using water and brine (2X). Organic layer with anhydrous sodium sulfate Dry, filter, concentrate on a rotary evaporator and dry. Heat solids Dissolved in diethyl ether and then added hexane to promote crystallization. This step affords analytically pure (R) -3- (1H-indole- 173.6 g of 3-yl) -2- (N-triphenylmethylamino) propanoic acid (0 . 389 mol) was isolated (two recovery yields 79% yield). FDMS446 (M⁺). C₃₀H₂₆N_TwoO_TwoAnalysis of:         Theory: C, 80.69; H, 5.87; N, 6.27.         Found: C, 80.47; H, 5.92; N, 6.10.   (B) (R) -3- (1H-indol-3-yl) -N- (2-methoxybenz) Production of (Zyl) -2- (N-triphenylmethylamino) propanamideCoupling   Anhydrous tetrahydrofuran (1.7 L) and anhydrous N, N -Hydroxybenzotriazole hydration in dimethylformamide (500 mL) (57.97 g, 0.429 mol), (R) -3- (1H-indole-3 -Yl) -2- (N-triphenylmethylamino) propanoic acid (179.8 g, 0 . 403 mol), and 2-methoxybenzylamine (56.0 mL, 0.4 29 mol) into a stirred solution of triethylamine (60.0 mL, 0.43 mol). 1) and 1- (3-dimethylaminopropyl) -3-ethoxycarbodiimide salt The acid salt (82.25 g, 0.429 mol) was added. Reduce mixture under nitrogen Warmed to room temperature for at least 20 hours. The mixture is concentrated on a rotary evaporator, Then redissolved in methylene chloride, 5% citric acid solution (2X), saturated sodium bicarbonate Aqueous solution (2X) and brine (2X). Organic layer anhydrous sulfur It was dried over sodium acid, concentrated on a rotary evaporator and dried. Next Recrystallize the desired product from hot ethyl acetate with analytically pure material 215. 8 g (0.381 mol, 95%) were obtained. FDMS 565 (M⁺). C₃₈H₃₅N_ThreeO_TwoAnalysis of:         Theory: C, 80.68; H, 6.24; N, 7.43.         Found: C, 80.65; H, 6.46; N, 7.50.   (C) (R) -3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) amino Production of 2- (N-triphenylmethylamino) propaneReduction of carbonyl   RED-AL® dissolved in anhydrous tetrahydrofuran (400 mL), [3 of sodium bis (2-methoxyethoxy) aluminum hydride in toluene . 4M solution] (535 mL, 1.819 mol) was added to anhydrous tetrahydrofuran under a nitrogen environment. (R) -3- (1H-indol-3-yl)-in (1.0 L) N- (2-methoxybenzyl) -2- (N-triphenylmethylamino) propanamide (228.6 g, 0.404 mol) was added to the refluxing solution using an addition funnel. The reaction mixture is purple It became a solution. Supersaturated Rochelle salt solution (potassium sodium tartrate tetrahydride) The reaction was stopped after at least 20 hours by slow addition. Isolate the organic layer , Washed with brine (2X), dried over anhydrous sodium sulfate, filtered, and rotary evaporated. Concentrated with a vaporizer to an oil. This product can be used without further purification Was used directly in the step.   (d) (R) -3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) -acetylamido Production of [no] -2- (N-triphenylmethylamino) propaneAcylation of secondary amines   (R) -3- (1H-Indoline in anhydrous tetrahydrofuran (1.2 L) at 0 ° C. under nitrogen environment -3-yl) -1- [N- (2-methoxybenzyl) amino] -2- (N-triphenylmethyla To a stirred solution of (mino) propane (0.404mol), triethylamine (66.5mL, 0.477mol) And acetic anhydride (45.0 mL, 0.477 mol). After 4 hours, mix the mixture on a rotary Concentrate with an evaporator, redissolve in methylene chloride and ethyl acetate, and add water (2 × ) And brine (2X), dried over anhydrous sodium sulfate, filtered and filtered. It was concentrated to a solid by a tally evaporator. The resulting solid is chloroform And flow through silica gel 60 (230-400 mesh) Elution was carried out with a 1: 1 mixture of ethyl acetate and hexane. The product is then treated with ethyl acetate Crystallized from a / hexane mixture. The resulting product, (R) -3- (1H-indole-3- Yl) -1- [N- (2-methoxybenzyl) acetylamino] -2- (N-triphenylmethyla Mino) propane was crystallized and 208.97 g of an analytically pure substance by three isolations (yield 87%). C₄₀H₃₉N_ThreeO_TwoAnalysis of:       Theory: C, 80.91; H, 6.62; N, 7.08.       Obtained: C, 81.00; H, 6.69; N, 6.94.   (e) (R) -2-amino-3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) ace Production of [Cylamino] propaneDeprotection   Formic acid (9.0 mL, 238.540 mmol) was added to a solution of (R) in anhydrous methylene chloride at 0 ° C. under a nitrogen environment. ) -3- (1H-Indol-3-yl) -1- [N- (2-methoxybenzyl) acetylamino] -2- (N -Triphenylmethylamino) propane (14.11 g, 23.763 mmol) Was. After 4 hours, the reaction mixture was concentrated on a rotary evaporator to an oil, Redissolved in diethyl ether and 1.0 N hydrochloric acid. Aqueous layer with diethyl ether And basified with sodium hydroxide to pH 12 or higher. Product Was extracted with methylene chloride (4X). Combine the organic extracts and dry with anhydrous sodium sulfate , Filtered and concentrated on a rotary evaporator to give a white foam. Compound (R) -2-amino-3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) a [Cetylamino] propane (7.52 g, 21.397 mmol) was isolated in 90% yield. More elaborate There was no need to make. (f) (R) -2-Amino-3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) acetyl [Ramino] propane dihydrochloride   (R) -3- (1H-Indol-3-yl) -1- [N- (2-methoxybenzoic acid) in 2 volumes of methylene chloride Stirred solution of (benzyl) acetylamino] -2- (N-triphenylmethylamino) propane Was cooled to −40 ° C. to −50 ° C. When the temperature of the reaction mixture is 0 ° C. Added at a rate that would not go above. The reaction mixture is stirred at 0 to 10 ° C for 30 minutes to 1 hour. Stirred.   To the reaction mixture was added 2 volumes of methyl t-butyl ether, and the resulting mixture Was stirred at 0 to 10 ° C for 30 minutes to 1 hour. Filter out the resulting crystalline solids And washed with methyl t-butyl ether. Reactive products are dried at 50 ° C under reduced pressure (Yield> 98%). C_{twenty one}H_{twenty five}N_ThreeO_TwoAnalysis of 2HCl:         Theory: C, 59.44; H, 6.41; N, 9.90.         Found: C, 60.40; H, 6.60; N, 9.99. (g) Production of potassium salt hydrate of 2-((4-cyclohexyl) piperazin-1-yl) acetic acid   Cyclohexylpiperazine (10.0 g, 0.059 mol) in 10 volumes of methylene chloride at room temperature Added. To this mixture was added sodium hydroxide (36 mL of a 2N solution, 0.072 mol) and bromide. Tetrabutylammonium (1.3 g, 0.004 mol) was added. Sodium hydroxide and After adding tetrabutylammonium bromide, methyl bromoacetate (7.0 mL, 0.073 mol) was added and the reaction mixture was stirred for 4-6 hours. Gas chromatog The progress of the reaction was monitored by luffy.   The organic fraction was separated and the aqueous phase was back-extracted with methylene chloride. Mix the organic phases and remove Washed twice with deionized water, once with saturated sodium bicarbonate solution and then with brine. Yes The organic phase was dried over magnesium sulfate, and the solvent was removed under reduced pressure to give methyl yellowish oil. 2- (4-Cyclohexyl) piperazin-1-yl) acetate was obtained.   Methyl 2- (4-cyclohexyl) piperazin-1-yl) acetate (10.0 g, 0.042 mol Was dissolved in 10 volumes of diethyl ether to give the title compound. The solution was cooled to 15 ° C. and then potassium trimethylsilanoate (5.9 g, 0.0 44) was added. The mixture was then stirred for 4-6 hours. Filter the reaction product Remove and wash twice with 5 volumes of diethyl ether, then 2 times with 5 volumes of hexane Washed and then dried in a vacuum oven at 50 ° C. for 12-24 hours. C₁₂H_{twenty one}KN_TwoO_Two・ 1.5H_TwoO:         Theory: C, 49.63; H, 7.98; N, 9.65.         Found: C, 49.54; H, 7.72; N, 9.11.   (h) (R) -2- [N- (2-((4-cyclohexyl) piperazin-1-yl) acetyl) amino] -3 -(1H-Indol-3-yl) -1- [N- (2-methoxybenzyl) acetylamino] propane Manufacturing of   Potassium 2-((4-cyclohexyl) piperazin-1-yl) acetate was added to anhydrous methyl chloride. The title compound was prepared by first cooling to a temperature of -8 ° C to -15 ° C in 5 volumes. Built. The mixture is added to isobutylchloroform at a rate such that the temperature does not exceed -8 ° C. Formate was added. The resulting reaction mixture was stirred for about 1 hour and the temperature was Maintained at 15 ° C.   Next, (R) -2-amino-3- (1H-indol-3-yl) -1- [N- (2-meth [Xybenzyl) acetylamino] propane dihydrochloride so that the temperature does not exceed 0 ° C Added at a great rate. The mixture is then charged with N-methylmorpholine at a temperature above 0 ° C. Added at an unbelievable rate. The mixture is then heated at a temperature between -15 ° C and -8 ° C for about Stir for 1 hour.   The reaction was stopped by adding 5 volumes of water. Wash the organic layer once with saturated sodium bicarbonate solution Was cleaned. Next, the organic phase was dried over anhydrous potassium carbonate and filtered to remove the desiccant. . Next, 2 equivalents of concentrated hydrochloric acid and then 1 volume of isopropyl alcohol were added to the filtrate. Next, methylene chloride was distilled under reduced pressure and exchanged with isopropyl alcohol.   Next, the final amount of isopropyl alcohol was concentrated to 3 volumes under reduced pressure. Reaction mixing The mass was cooled to 20-25 ° C and the product was allowed to crystallize for at least 1 hour. Next, The desired product is recovered by filtration, washed with plenty of isopropyl alcohol and colorless. A filtrate was obtained. The crystal cake was dried at 50 ° C. under reduced pressure. MS 560 (M + 1⁺). C₃₃H₄₅N_FiveO_ThreeAnalysis of:         Theory: C, 70.81; H, 8.10; N, 12.51.         Found: C, 70.71; H, 8.21; N, 12.42.   (R) -3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) acetylamino]- 2- [N- (2- (4- (piperidin-1-yl) piperidin-1-yl) acetyl) amino] propane Synthesis of (a) 2- (4- (piperidin-1-yl) piperidin-1-yl) acetic acid, potassium salt   4- (piperidin-1-yl) piperidine (1.20 kg, 7.13 mol) Added to styrene (12.0 L). Then, tetrabutylammonium bromide (0.150 kg, 0.47 mol) and sodium hydroxide (1.7 L of a 5N solution, 8.5 mol). Reaction mixture Was cooled to 10 to 15 ° C., and methyl bromoacetate (1.17 kg, 7.65 mol) was added. The resulting solution was stirred for a minimum of 16 hours.   Next, deionized water (1.2 L) was added to the mixture and the phases were separated. Methyle chloride in aqueous layer (2.4 L). Combine the organic fractions, deionized water (3x 1.2L), saturated bicarbonate Washed with sodium solution (1.1 L) and saturated sodium chloride solution (1.1 L). next, The organic fraction was dried over anhydrous magnesium sulfate and concentrated on a rotary evaporator. To give an oil, methyl 2- (4- (piperidin-1-yl) piperidin-1-yl) acetate 1.613 kg (93.5%) was obtained.   Methyl 2- (4- (piperidin-1-yl) piperidin-1-yl) a in methanol (2.4 L) A solution of acetate (2.395 kg, 9.96 mol) in methanol (10.5 L) under a nitrogen environment It was added to a solution of potassium hydroxide (0.662 kg, 10.0 mol ＠ purity 85%). The reaction mixture Heated to 45-50 ° C for a minimum of 16 hours.   The solvent of the solution was changed from methanol to acetone (15.0 L) using a rotary evaporator. ) Was replaced. The solution was cooled slowly to room temperature over 16 hours. The solid obtained The material was filtered, rinsed with acetone (5.0 L) and then dried to give 2- (4- (piperidine-1 2.471 kg (93.8%) of -yl) piperidin-1-yl) acetic acid / potassium salt were obtained. MS 265 (M⁺¹).   (B) (R) -3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) acetyla Mino] -2- [N- (2- (4-piperidin-1-yl) piperidin-1-yl) acetyl) amino] p Production of Lopan   (R) -2-amino-3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) acetyl [Amino] propane dihydrochloride (50.0 g, 0.118 mol) in methylene chloride under nitrogen The title compound was prepared by first mixing with 100 mL.   In a second flask under a nitrogen atmosphere, 2- (4- (piperidin-1-yl) piperidin-1-y L) Potassium acetate (62.3 g, 0.236 mol) was added to 600 mL of methylene chloride. This mixture The material was cooled to about -10 ° C and stirring was continued. 2- (4- (piperidin-1-yl) piperidin-1- (Il) To prevent the temperature of the potassium acetate salt mixture from increasing appreciably, Isobutyl chloroformate (23 mL, 0.177 mol) was added dropwise to the mixture.   The reaction mixture was stirred at about −10 ° C. for about 1.5 hours, at which point it was prepared earlier (R) -2-amino-3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) acetyl Amino] propane dihydrochloride / methylene chloride mixture was converted to 2- (4-piperidin-1-yl) pi Peridin-1-yl) acetic acid potassium salt / isobutyl chloroformate / methylene chloride Added slowly to the solution. Next, the obtained solution is heated at a temperature of -15 ° C to -8 ° C for about 1 hour. Stirred.   Remove the reaction mixture from the ice bath, warm to 15-20 ° C, and stop the reaction by adding 200 mL of water. I did. The pH of the solution was adjusted to 2.3-2.7 by adding 1N sulfuric acid. Separate the layers and separate the aqueous layer Washed with 100 mL of methylene chloride.   The organic fractions were combined and washed with water (100mL). After washing with water, methylene chloride ( (500 mL) and mixed with the aqueous fraction obtained above. Methylene chloride (500 mL) Add to the combined aqueous layers, stir the mixture at room temperature for 15 minutes, and add It was basified to a final pH of 9.8-10.2.   The organic and aqueous fractions were separated. The aqueous fraction was washed with methylene chloride to form the organic fraction. Methylene chloride was added. Next, the organic fraction was washed with a saturated sodium bicarbonate solution (100 m L) and water (50 mL). Bicarbonate wash from organic fraction Separate and back extract with methylene chloride (50 mL). Methylene chloride fractionation of the back extract And the combined fractions were dried over magnesium sulfate. Filter magnesium sulfate And the volatiles were removed by distillation under reduced pressure to give the title product as a foam (72.5 g, Yield> 98%). MS 559 (M⁺¹) C₃₃H₄₅N_FiveO_ThreeAnalysis of:         Theory: C, 70.81; H, 8.10; N, 12.51.         Found: C, 70.57; H, 8.05; N, 12.39.   Another method for preparing compounds of formula I is as follows. (R) -3- (1H-indol-3-yl) -2- (N-triphenylmethylamino) propanoic acid, N -Methylmorpholine salt (N-trityl-D-tryptophan N-methylmorpholine salt) Manufacturing.   1L4 neck with stirrer, condenser, probe and stopper In a flask, D-tryptophan (40.0 g, 0.196 mol), acetonitrile (240 mL) , And 1,1,1,3,3,3-hexamethyldisilazane (39.5 g, 0.245 mol) were added. Profit The resulting mixture was heated to 50-60 <0> C and stirred until homogeneous. In another beaker Slurry trityl chloride (60.06 g, 0.215 mol) and acetonitrile (120 mL) I made it. Add the slurry to the silylated tryptophan mixture and place the beaker in acetonitrile. Rinse with 40 mL of Toril. Add N-methylmorpholine (23.7 mL, 21.8 g, 0.21 g) to the reaction mixture. 6 mol) was added and the resulting mixture was stirred for 1 hour. Chromatograph the progress of the reaction I monitored it.   After sufficient progress, water (240 mL) was added dropwise to the reaction mixture, and the resulting mixture was The mixture was cooled to below ℃, stirred for 30 minutes and filtered. Wash the residue with water and then dry 108.15 g (> 99% yield) of the desired title product were obtained. C₃₀H₂₆N_TwoO_TwoAnalysis of:         Theory: C, 80.69; H, 5.87; N, 6.27.         Found: C, 80.47; H, 5.92; N, 6.10. (R) -3- (1H-indol-3-yl) -N- (2-methoxybenzyl) -2- (N-triphenylmeth Production of (tylamino) propanamide   2 with stirrer, condenser, and thermocouple (thermocouple) L In a four-necked flask, add N-trityl-D-tryptophan N-methylmorpho Phosphate (108.0 g, 0.196 mol), acetonitrile (800 mL), 2-chloro-4,6-dimethoxy -1,3,5-triazine (38.63 g, 0.22 mol) and N-methylmorpholine (29.1 mL) were added. I got it. The resulting mixture was stirred at ambient temperature until homogeneous (about 10 minutes).   After about 1 hour, 2-methoxybenzylamine (29 mL) was added. 35 of the resulting mixture Heated to ° C. and maintained at that temperature overnight. Monitor the progress of the reaction by chromatography. I did it. Next, water (750 mL) was added to the reaction mixture, and the resulting mixture was cooled to 10 ° C or lower. And stirred for 30 minutes, then filtered. Wash the residue with water (about 100 mL) And dried to give the desired title product (yield 87% and 91% in two operations) ). FDMS 565 (M⁺). C₃₈H₃₅N_ThreeO_TwoAnalysis of:         Theory: C, 80.68; H, 6.24; N, 7.43.         Found: C, 80.65; H, 6.46; N, 7.50.Reduction of carbonyl (R) -3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) amino] -2- (N-tri Production of (phenylmethylamino) propane   RED-AL® dissolved in anhydrous tetrahydrofuran (400 mL), [3 of sodium bis (2-methoxyethoxy) aluminum hydride in toluene . 4M solution] (535 mL, 1.819 mol) was added to anhydrous tetrahydrofuran under a nitrogen environment. In (1.0 L), the previously formed acylated product, (R) -3- (1H-indol-3-yl) -N- ( 2-methoxybenzyl) -2- (N-triphenylmethylamino) propanamide (228.6 g, 0.404 mol) was added to the refluxing solution using an addition funnel. Reaction mixture purple A color solution resulted. Supersaturated Rochelle salt solution (potassium sodium tartrate tetrahydrogen The reaction was stopped after at least 20 hours by slowly adding Organic layer Isolate, wash with brine (2X), dry over anhydrous sodium sulfate, filter, and It was concentrated to an oil by a reevaporator. Further purification of this product And used directly in the next step.Acylation of secondary amines   (R) -3- (1H-Indol-3-yl) -1- [N- (2-methoxybenzyl) -acetylamino] Production of 2- (N-triphenylmethylamino) propane   (R) -3- (1H-indole in anhydrous tetrahydrofuran (1.2 L) under a nitrogen atmosphere at 0 ° C. Ru-3-yl) -1- [N- (2-methoxybenzyl) amino] -2- (N-triphenylmethylamido (N) In a stirred solution of propane (0.404 mol), triethylamine (66.5 mL, 0.477 mol) and And acetic anhydride (45.0 mL, 0.477 mol) were added. After 4 hours, the mixture was Concentrate with a evaporator, redissolve in methylene chloride and ethyl acetate, add water (2X) And brine (2X), dried over anhydrous sodium sulfate, filtered, and The solid was concentrated by a reevaporator. The obtained solid is converted into chloroform. Dissolve, pour over silica gel 60 (230-400 mesh), add ethyl acetate and Eluted with a 1: 1 mixture of sun. The product is then added to an ethyl acetate / hexane mixture Crystallized from The resulting product, (R) -3- (1H-indol-3-yl) -1- [N- (2-meth Toxibenzyl) acetylamino] -2- (N-triphenylmethylamino) propane Crystallized and three isolations gave 208.97 g (87% yield) of analytically pure material. . C₄₀H₃₉N_ThreeO_TwoAnalysis of:       Theory: C, 80.91; H, 6.62; N, 7.08.       Obtained: C, 81.00; H, 6.69; N, 6.94.Deprotection (R) -2-amino-3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) acetyl Production of [amino] propane dihydrochloride   (R) -3- (1H-Indol-3-yl) -1- [N- (2-methoxybenzoic acid) in 2 volumes of methylene chloride Stirred solution of (benzyl) acetylamino] -2- (N-triphenylmethylamino) propane Was cooled to −40 ° C. to −50 ° C. When the temperature of the reaction mixture is 0 ° C. Added at a rate that would not go above. The reaction mixture is stirred at 0 to 10 ° C for 30 minutes to 1 hour. Stirred.   To the reaction mixture was added 2 volumes of methyl t-butyl ether, and the resulting mixture Was stirred at 0 to 10 ° C for 30 minutes to 1 hour. Filter out the resulting crystalline solids And washed with methyl t-butyl ether. Reactive products are dried at 50 ° C under reduced pressure (Yield> 98%). C_{twenty one}H_{twenty five}N_ThreeO_TwoAnalysis of 2HCl:         Theory: C, 59.44; H, 6.41; N, 9.90.         Found: C, 60.40; H, 6.60; N, 9.99. Preparation of 2-((4-cyclohexyl) piperazin-1-yl) acetic acid potassium salt hydrate   Cyclohexylpiperazine (10.0 g, 0.059 mol) in 10 volumes of methylene chloride at room temperature Added. To this mixture was added sodium hydroxide (36 mL of a 2N solution, 0.072 mol) and bromide. Tetrabutylammonium (1.3 g, 0.004 mol) was added. Sodium hydroxide and After addition of tetrabutylammonium bromide, methyl bromoacetate (7.0 mL, 0.1 mL) was added. 073 mol) was added and the reaction mixture was stirred for 4-6 hours. Gas chromatograph The progress of the reaction was monitored graphically.   The organic fraction was separated and the aqueous phase was back-extracted with methylene chloride. Mix the organic phases and remove Washed twice with deionized water, once with saturated sodium bicarbonate solution and then with brine. Yes The organic phase was dried over magnesium sulfate, and the solvent was removed under reduced pressure to give methyl yellowish oil. 2- (4-Cyclohexyl) piperazin-1-yl) acetate was obtained.   Methyl 2- (4-cyclohexyl) piperazin-1-yl) acetate (10.0 g, 0.042 mol Was dissolved in 10 volumes of diethyl ether to give the title compound. The solution was cooled to 15 ° C. and then potassium trimethylsilanoate (5.9 g, 0.0 44) was added. The mixture was then stirred for 4-6 hours. Filter the reaction product Remove and wash twice with 5 volumes of diethyl ether, then 2 times with 5 volumes of hexane Washed and then dried in a vacuum oven at 50 ° C. for 12-24 hours. C₁₂H_{twenty one}KN_TwoO_Two・ 1.5H_TwoO:         Theory: C, 49.63; H, 7.98; N, 9.65.         Found: C, 49.54; H, 7.72; N, 9.11. (R) -2- [N- (2-((4-cyclohexyl) piperazin-1-yl) acetyl) amino] -3- (1H- Production of indole-3-yl) -1- [N- (2-methoxybenzyl) acetylamino] propane Construction   Potassium 2-((4-cyclohexyl) piperazin-1-yl) acetate was added to anhydrous methyl chloride. The title compound was prepared by first cooling to a temperature of -8 ° C to -15 ° C in 5 volumes. Built. The mixture is added to isobutylchloroform at a rate such that the temperature does not exceed -8 ° C. Formate was added. The resulting reaction mixture was stirred for about one hour and the temperature was Maintained at 15 ° C.   Next, (R) -2-amino-3- (1H-indol-3-yl) -1- [N- (2-meth [Xybenzyl) acetylamino] propane dihydrochloride so that the temperature does not exceed 0 ° C Added at a great rate. The mixture is then charged with N-methylmorpholine at a temperature above 0 ° C. Not at such a speed. The mixture is then brought to a temperature of -15 ° C to -8 ° C for about 1 hour. Stirred for hours.   The reaction was stopped by adding 5 volumes of water. Wash the organic layer once with saturated sodium bicarbonate solution Was cleaned. Next, the organic phase was dried over anhydrous potassium carbonate and filtered to remove the desiccant. . Next, 2 equivalents of concentrated hydrochloric acid and then 1 volume of isopropyl alcohol were added to the filtrate. Next, methylene chloride was distilled under reduced pressure and exchanged with isopropyl alcohol.   Next, the final amount of isopropyl alcohol was concentrated to 3 volumes under reduced pressure. Reaction mixing The mass was cooled to 20-25 ° C and the product was allowed to crystallize for at least 1 hour. Next, The desired product is recovered by filtration, washed with plenty of isopropyl alcohol and colorless. A filtrate was obtained. The crystal cake was dried at 50 ° C. under reduced pressure. MS 560 (M + 1⁺). C₃₃H₄₅N_FiveO_ThreeAnalysis of:         Theory: C, 70.81; H, 8.10; N, 12.51.         Found: C, 70.71; H, 8.21; N, 12.42. Production of 2- (4- (piperidin-1-yl) piperidin-1-yl) acetic acid, potassium salt  4- (piperidin-1-yl) piperidine (1.20 kg, 7.13 mol) Added to styrene (12.0 L). Then, tetrabutylammonium bromide (0.150 kg, 0.47 mol) and sodium hydroxide (1.7 L of a 5N solution, 8.5 mol). Reaction mixture Was cooled to 10 to 15 ° C., and methyl bromoacetate (1.17 kg, 7.65 mol) was added. The resulting solution was stirred for a minimum of 16 hours.   Next, deionized water (1.2 L) was added to the mixture and the phases were separated. Methyle chloride in aqueous layer (2.4 L). Combine the organic fractions, deionized water (3x 1.2L), saturated bicarbonate Washed with sodium solution (1.1 L) and saturated sodium chloride solution (1.1 L). next, The organic fraction was dried over anhydrous magnesium sulfate and concentrated on a rotary evaporator. To give an oil, methyl 2- (4- (piperidin-1-yl) piperidin-1-yl) a 1.613 kg (93.5%) of the acetate was obtained.   Methyl 2- (4- (piperidin-1-yl) piperidin-1-yl) a in methanol (2.4 L) A solution of acetate (2.395 kg, 9.96 mol) in methanol (10.5 L) under a nitrogen environment It was added to a solution of potassium hydroxide (0.662 kg, 10.0 mol ＠ purity 85%). The reaction mixture Heated to 45-50 ° C for a minimum of 16 hours.   The solvent of the solution was changed from methanol to acetone (15.0 L) using a rotary evaporator. ) Was replaced. The solution was cooled slowly to room temperature over 16 hours. The solid obtained The material was filtered, rinsed with acetone (5.0 L) and then dried to give 2- (4- (piperidine- 2.471 kg (93.8%) of 1-yl) piperidin-1-yl) acetic acid / potassium salt was obtained. MS 265 (M⁺¹). (R) -3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) acetylamino] -2- [N- (2- (4-piperidin-1-yl) piperidin-1-yl) acetyl) amino] propane Manufacture of dihydrochloride and trihydrate   Under a nitrogen environment, 2- (4-piperidin-1-yl) piperidin-1-yl) acetic acid / potassium Salt (0.75 kg, 2.84 mol) was added to methylene chloride (7.5 L). The resulting mixture is -15 Cool to ~ -8 ° C and add isobutyl chloroformate (0.29 kg, 2.12 mol) to the reaction mixture. Was added at such a rate that the temperature was maintained below -8 ° C. After addition, the resulting reaction mixture The mixture was stirred at -15 to -8C for 90 minutes.   The reaction mixture was then cooled to -35 ° C and solid (R) -2-amino-3- (1H-indole Ru-3-yl) -1- [N- (2-methoxybenzyl) amino] propane dihydrochloride (0.60 kg, 1.14 mol) was added at such a rate that the reaction temperature was kept below -20 ° C. After addition The reaction mixture was stirred for about 1 hour while maintaining the temperature at -37 to -20C. The reaction was stopped by adding deionized water (7.5 L). The reaction mixture was added with 5N sodium hydroxide. And basified to pH 12.8-13.2. The aqueous fraction was removed and retained. Add deionized water (3.75 L) to the organic fraction and add enough 5N sodium hydroxide. The pH was readjusted to 12.8-13.2.   The two aqueous fractions are combined, back-extracted with methylene chloride (1.5 L) and then the aqueous fraction Abandoned the painting. The organic fractions were combined and washed with deionized water (4x3.5L). The extraction The effluents were combined, back-extracted with methylene chloride (1.5 L) and then discarded. Two organic layers And washed with saturated sodium chloride solution (3.7 L).   The organic fraction was dried over anhydrous magnesium sulfate, filtered, and rotary evaporated. The solvent was exchanged from methylene chloride to acetone (3.75 L) with a mouse. Hydrochloric acid (6N solution 0.48 L, 2.88 mol) of an aqueous solution and seed crystals (2 g) were added and the mixture was Stir for 0-90 minutes. Next, acetone (13.2 L) was added, and the slurry was stirred for 1 hour. While stirring. Next, the obtained solid is filtered and washed with acetone (2 × 1.4 L). And dried to give (R) -3- (1-indol-3-yl) -1- [N- (2-methoxybenzyl) a Cetylamino] -2- [N- (2- (4- (piperidin-1-yl) piperidin-1-yl) acetyl) 633 g (90%) of [amino] propane dihydrochloride trihydrate were obtained. (R) -3- (1H-Indol-3-yl) -1- [N- (2-methoxybenzyl) acetylamino]- 2- [N- (2- (4- (piperidin-1-yl) piperidin-1-yl) acetyl) amino] propane ・ Manufacture of dioxalate   In a 500 mL jacketed round bottom flask, add 2- (4- (piperidin-1-yl) piperidi 1-yl) acetic acid, potassium salt (25.0 g, 94.5 mmol) and N, N-dimethyl 375 mL of formamide was charged. The resulting slurry is cooled to -19 ° C and Til chloroformate (12.9 g, 94.5 mmol) was added over 5 minutes. Profit The resulting mixture was stirred for 20 minutes, then 75 mL of anhydrous N, N-dimethylformamide (R) -2-amino-3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) dissolved in L) acetylamino] propane dihydrochloride (25.0 g, 58.1 mmol) for 10 minutes Added over.   Next, the resulting mixture was cooled to 0 ° C., stirred for about 10 minutes, and then warmed to room temperature. Was. The progress of the reaction was monitored by chromatography. Reactant conversion after 90 minutes Was 99% by high performance liquid chromatography.   Ethyl acetate (375 mL) and saturated sodium bicarbonate solution (375 mL) ). The aqueous layer was back-extracted with 375 mL of ethyl acetate. Mix the organic fractions, Washed with water (3 × 375 mL) and dried over magnesium sulfate. Next, Lithium is added to the previously obtained aqueous fraction and the resulting basic solution is extracted with ethyl acetate. Next, the organic fraction is dried with magnesium sulfate.   Next, the mixed dried organic fraction is treated with a concentrated oxalic acid solution. Filter the resulting solid, Drying at 50 ° C. in a vacuum oven gave 23.5 g of the desired intermediate.   As one of ordinary skill in the art will appreciate, the mixed anhydride step is performed using the anhydrous N, N-dimethyl It will work in many organic solvents in addition to formamide. Solvents that can be used Representative examples include acetonitrile, tetrahydrofuran, and dichloromethane. I will. The mixed acid anhydride step can be performed at a temperature of 0 ° C. or less.   The oxalate is ethyl acetate, and possibly acetone, acetonitrile and t-butyl. It can be isolated from other solvents including tyl methyl ether. However, The use of oxalic acid in precipitation is very heavy since many acids do not produce precipitates. It is important. Of the acids tried, those found not to be sufficient for the method of the invention are: Citric acid, anhydrous hydrochloric acid, tartaric acid, mandelic acid, trifluoroacetic acid, p-nitrobenzo Perfume acid, phenoxyacetic acid, maleic acid, fumaric acid, glutaric acid, adipic acid, meta Sulfonic acid, p-toluenesulfonic acid, pamoic acid, tiger -1,2-cyclohexanedicarboxylic acid, succinic acid, phthalic acid, trans-1,2-di Aminocyclohexane-N, N, N ', N'-naphthalenedisulfonic acid, and 5-sulfosa Lithilic acid. Only oxalic acid and 1,5-naphthalenedisulfonic acid are solids with reproducibility Occurred. (R) -3- (1H-Indol-3-yl) -1- [N- (2-methoxybenzyl) acetylamino]- 2- [N- (2- (4-piperidin-1-yl) piperidin-1-yl) acetyl] amino] propane ・ Manufacture of dihydrochloride and trihydrate   In a large beaker, add (R) -3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) a Cetylamino] -2- [N- (2- (4-piperidin-1-yl) piperidin-1-yl) acetyl) a Mino] propane dioxalate (13.4 g, 18.1 mmol), methylene chloride (58.16 mL, 78.51 g ) And water (118.59 mL) were added. The resulting mixture is stirred and the pH of the reaction mixture is Was adjusted to 10-12 using 50% caustic.   The layers were separated and the organic phase was back-extracted with water (101.44 mL). Organic fraction with jacket The mixture was transferred to a round bottom flask, and the solvent was exchanged using about 23 volumes of acetone. Aceto Portion was added to the product solution and the added amount was distilled off. Progress of solvent exchange Was monitored by NMR. The amount of desired product is determined by high performance liquid chromatography. Monitored.   Sufficient water is added to bring the water concentration to 11% and the resulting mixture is heated to 55 ° C. Was. The pH was lowered to 2.0 by adding a sufficient amount of concentrated hydrochloric acid, and then the reaction mixture was allowed to stand for 45 minutes. And cooled to 37 ° C.   The product solution was seeded and stirred for 10-30 minutes. The product solution over 2 hours After cooling to 19 ° C., acetone (10 volumes) was added over 3 hours, after which the temperature was raised to 1 The reaction mixture was stirred at 1-3 ° C. for 1-3 hours. The product solution is filtered and the remaining The material was washed with 6.67 volumes of acetone. The residue is then placed in a vacuum oven for 42 hours. Drying at <RTIgt; 0 C </ RTI> provided the desired title product.   Other compounds of the formula I are prepared substantially according to the above and using the corresponding starting materials. Could be.   Of compounds considered to be effective as tachykinin receptor antagonists Biological efficacy is tested against known NK-1 and NK-2 receptor sites Use an initial screening assay to quickly and accurately measure compound binding This can be confirmed by Tachykinin receptor antagonists reviewed Assays useful for titration are well known in the art (J. Jukki et al., Life Sciences, 49: 1463-1469 (1991); N. Kucharczyk et al., Journal of Medicinal. Chemistry, 36: 1654-1661 (1993); N. Louisi et al., Biochemical and Biophysical. Research Communications, 176: 894-901 (1991)).NK-1 receptor binding assay   Radioreceptor binding assay using a modification of a previously published protocol (D.G. Payan et al., Journal of Immunology, 133: 3260-3265 (1984)). This In the assay, a partial preparation of IM9 cells (1 × 10⁶Pieces / tube, 1 0% fetal calf serum in RPMI 1604 medium) in the presence of increasing competitor concentrations. In the presence of 20 pM at 4 ° C. for 45 minutes¹²⁵Incubation with I-labeled substance P Was done.   The IM9 cell line is a well-characterized cell line that is readily available to the public ( For example, Annals of the New York Academy of Science, 190: 221-234 (1972); Nature (London), 251: 443-444 (1974); Proceedings of the National Academy of Science (USA), 71: 84-88 (1974)). The cells were 50 μg / mL gentamic sulfate. Cultured routinely in RPMI 1640 supplemented with Syn and 10% fetal calf serum.   Using a filter pre-soaked in 0.1% polyethyleneimine for 20 minutes The reaction is terminated by filtration with a glass fiber filter collection system. Was. Specific binding of labeled substance P was measured in the presence of 20 nM unlabeled ligand. Was.   The various compounds used in the methods of the present invention are effective NK-2 receptor antagonists. He is also a gonist.NK-2 receptor binding assay   CHO-h, a CHO-derived cell line transformed with the human NK-2 receptor NK-2R cells (expressing about 400,000 such receptors per cell ) Is 75 cm in 10% fetal calf serum-supplied minimum essential medium (Alpha modified method)^TwoH Grow in Lasco or roller bottles. Human NK-2 receptor gene The sequence is described in N.P.Gerard et al., Journal of Biological Chemistry, 265: 20455-20462 (1990 ).   Prepare 30 confluent roller bottle cultures for membrane preparation. Place each roller bottle in calcium- and magnesium-free Dulbeccolin Wash with 10 mL of acid-buffered saline (PBS), and then remove the enzyme-free cell separation solution ( PBS base, obtained from Specialty Media, Inc.) Released. After an additional 15 minutes, the separated cells were pooled and 1000 RPM in a clinical centrifuge. For 10 minutes. The membrane was prepared by transferring the cell pellet to 50 mM Tris buffer (pH 7.0). 4) 10-15 seconds with Tekmar® homogenizer in 300 mL Homogenized and then 12000 RPM using a Becman JA-14® rotor. It was prepared by centrifugation at (20,000 xg) for 30 seconds. Pellet the above steps Wash once and wash the final pellet with 50 mM Tris buffer (pH 7.4) 100 Resuspended at ~ 120 mL and aliquots of 4 mL were stored frozen at -70 ° C. This key The protein concentration of the product was 2 mg / mL.   For the receptor binding assay, a 4 mL sub-label of the CHO-hNK-2R membrane preparation was Transfer the book to 50 mM Tris (pH 7.4), 3 mM magnesium chloride, 0.02% bovine. Assay containing serum albumin (BSA) and 4 μg / mL chymostatin Suspended in 40 mL of buffer. Homogenate (40 μg protein) per sample 2 A 00 μL volume was used. The radioactive ligand is [¹²⁵I] Iodohistidyl-new Rokinin A (New England Nuclear, NEX-252), 2200 Ci / mmol . The ligand was prepared at 20 nCi / 100 μL in assay buffer and assayed. The final concentration in was 20 pM. Non-specific binding using 1 μM eledoisin It was measured. Standard concentration-response curves for 10 concentrations of eledoisin from 0.1 to 1000 nM It was used for.   All samples and standards were screened (one dose) for dimethyl sulfoxide Cid (DMSO) in 10μ, IC₅₀For measurement, incubate in 5 μL of DMSO. Added to the solution. The order of additives for incubation is Buffer 190 or 195 μL, homogenate 200 μL, sample 10 in DMSO Or 5 μL and 100 μL of radioactive ligand. Inject sample at room temperature for 1 hour Cultivation, and then with cell harvester, 0.5% BS Filter with a filter immersed for 2 hours in 50 mM Tris buffer (pH 7.7) containing A I have. Wash the filter three times with about 3 mL of cold 50 mM Tris buffer (pH 7.7) Was cleaned. Next, the filter was punched out into a circular shape to obtain a 12 x 75 mm polystyrene chip. The sample was placed in a tube and counted with a gamma counter.   The method of the present invention is directed to a mammal having symptoms of interstitial cystitis and / or urethral syndrome. It has been found to be effective in treating physical, especially middle-aged women. In this regard The clinical and local immune responses to the compounds of the present invention were Diagnosed according to agreed criteria developed in 1987 at the Shop An open-trail study will be conducted on 10 women with interstitial cystitis. Patient's condition Because of the status and clinical response, we (urgency), nocturia, dysuria, and suprapubic pain in US Pat. No. 5,859 (issued on September 8, 1992) (the contents of which are incorporated herein by reference. Score) (scale 0-2). The compound of the present invention It is administered as a single daily dose as determined by a volume titration test. Cellular inflammation marker Urine interleukin-2 inhibitory activity (IL-2-IN) It was measured using a tarleukin-2-dependent cell line.   A decrease in the patient's clinical symptoms was observed. Side effects of the drug were minimal. Before treatment The presence of cellular inflammation was confirmed by urinary IL-2-IN activity, and IL- 2-IN activity is normal in most patients regardless of the severity of the condition, Indicates that the compound of formula I has an immunosuppressive effect. Data are for compounds of formula I It suggests that it may be an effective and convenient medicine that is well tolerated in treating cystitis.   Furthermore, as shown more clearly in Example 2 below, the present inventors treated urethral syndrome. A similar response has been observed for treatment. As a result, the test data is Compound used is effective for treating interstitial cystitis and / or urethral syndrome This clearly indicates that it may be a therapeutic agent.   As a result, the compounds of formula I not only provide effective relief but also Interstitial cystitis and / or urine because they can be used and are relatively inexpensive It has been found to be particularly suitable for the treatment of tract syndrome. Administered a compound of formula I Patients have substantially reduced morbidity exhibited by the two types of painful bladder disorders. To maintain the patient's daily activity in a relatively normal state of living compared to the state before treatment. I knew I could do it.   The present invention is further described by the following non-limiting examples.Example 1 Materials and methods patient. National Institutes of Health Workshop on Interstitial Cystitis (August 1987) Mon, Gillenwater, J.Y. and Wein, A.J .: Summary of the National Institute of  Arthritis, Diabetes, Digestive and Kidney Diseases Workshop on Interstitial Cystitis, National Institutes of Health, Bethesda, Md., Aug. 28-29, 1987, J. Urol., 140: 203, 1988) Diagnosis of interstitial cystitis according to US Patent No. 5,145,859, Assign 10 51-year-old women.Interstitial cystitis: diagnostic criteria Inclusion criteria Exclusion criteria Hunner ulcer (automatically wraps if present under 18 years old) Benign or malignant tumor                                      Irradiation, tuberculosis, bacterial Positive factors (at least to include or cyclophosphamide cystitis 2): vaginitis                                      Duration of symptoms <1 year Suprapubic pain, pelvic pain, urethral pain, vaginal pain Gynecological cancer Or perineal pain urethral diverticulum, bladder                                      Or lower urethral stones Bladder distention (water pressure 80 cm x 1 mi active herpes (HSV II) n. ) Glomerular invasion during cystoscopy after awakening frequency <12 at 12 hours Glomerulation Nocturnal polyuria <2                                      Nervous bladder dysfunction Cyst Metrogram Awakening capacity> 400 mL, Compliance in the cystometrogram Lack of urgency due to bladder filling Lower symptoms relieved by antibiotics, Bladder filling pain relieved by emptying Urinary analgesics or fungicides   Perform cystometrics after discontinuing other treatments and before setting treatment The awake bladder volume of all patients was less than 350 mL (range 150 mL to 340 mL).   Symptom evaluation: The symptom score (total score range: 0 to 10) is the basis of the evaluation of the therapeutic effect. It is the cornerstone. A numerical value is assigned to the severity of each symptom as follows. Investigation of symptom severity   At the time of diagnosis and before treatment, the package of criteria (above) agreed at the NIH workshop Patients that meet the inclusion or exclusion descriptor parameters have at least Scoring to “4” (frequent urine <1; urgency <1; nocturia <1; and voiding Dysuria or suprapubic pain <1).   Urine collection: Collect urine specimens from all patients before and during treatment. Urinated The urine is centrifuged at 1000 × g at 4 ° C. for 10 minutes, and the supernatant is separated from the precipitate . The urine supernatant is subjected to 0.2μ filtration (cellulose acetate) at 4 ° C to remove any bacteria. Remove 1 mL of aliquots, remove trash, and measure creatinine (CREATINI NE II ANALYZER®, Beckman Instruments, Inc., Brea, California). The supernatant is filtered using a filtration device (5000 MW cutoff, Amicon, Deavers, Massachusetts). 0.1 μg / mL albumin (Sigma, St. Louis, Missouri) -containing phosphate buffer Ultrafiltrate in opposing saline (PBS) at 3x volume. 3500 MW of concentrated supernatant Dialyze using a cut-off tube, freeze the shell on dry ice, and freeze under vacuum. Let dry. Store the powder at -20 <0> C.   Measurement of IL-2-IN activity: A bioassay for IL-2-IN was provided by Gillis et al. The method for measuring IL-2 activity described above is partially modified (S. Gillis et al., T-Cell Growth Factor: Parameters of Production and a Quantitative Microassay for Activity, Journal of Immunology, 120: 2027 (1978)). Murine IL-2 dependent cells A cytotoxic T cell line (CTLL-N) is derived from the CT-6 cell line (J. Kusugami et al.). "Intestinal Immune Reactivity to Interleukin-2 Differs Among Crohn's Disease, Ulcerative Colitis and Controls, Gastroenterology, 97: l (1989)) . CTLL-N is 2.9 mg / mL glucose, 9.4 mM HEPES buffer 1.9 mg / mL glutamine, 289 μg / mL arginine, 0.12 M Sub amino acid, 5x10^-FiveM2-mercaptoethanol, 4.5% fetal bovine serum, 90 units / mL penicillin, 90 μg / mL streptomycin, 22 μg / m L fungizone, 0.45 mg / mL gentamicin and 20 units / mL human Dulbecco's modified Eagle's medium supplemented with recombinant IL-2 (DMEM; 4.5 g / L glucose) medium and Roswell Park Memorial Institute (RPMI) 1640-1 : 1 mixture and maintained in liquid culture.   After washing the CTLL-N, the culture solution was^-FiveResuspended at 1 / mL. The assay is Perform in triplicate as follows. Serial dilutions of sample aliquots (50 μL) , A 1:10 dilution of human recombinant IL-2 standard and 10^-FourCTLL-N (100 μL) into a microtiter well. Humidify the microtiter plate 6 % CO_TwoIncubate in air at 37 ° C. for 24 hours, and at 19 hours 1 μCi / well of methyltritiated thymidine (specific activity 6.7 Ci / mM, Ne w England Nuclear, IE. (Dupont, Boston, Massachusetts).   Cells are collected on glass filter discs. Disc in scintillation fluid And thymidine incorporation is measured with a liquid scintillation spectrophotometer. IL -2 Inhibitory activity is calculated by a modified probit assay.   The “maximum” growth was determined by the amount of exogenous IL-2 activity in the control μL wells. Incorporation of tritiated thymidine (four times for each assay) Evaluation). Proliferation "minimum" is the percentage of growth caused by the IL-2 inhibitor standard. Obtained from the lowest uptake of lithiated thymidine. Control the calculated value of the probit Correct for small interassay variations in thymidine incorporation in roll wells A comparison between assays of inhibitor activity between urine samples was performed. By this data processing, the assay of the calculated value of the IL-2 inhibitory activity in the lyophilized urine sample was performed. The variation for each item is 10% or less. IL-2-IN activity is unit / mg urine creatine It is expressed in nin (U / mg uc). IL-2-IN activity is urine in healthy adults At 0.05 U / mgu. c. (J. Fleischmann et al., Journal of Biological Regulators and Homeostatic Agents, 4:73 (1990)).   Drug therapy assignment: All patients are initially treated with a total daily dose of 30 mg ( Administered as a single extended release tablet).   Patient monitoring: 1 for the first 2 months of treatment and 1 for 2 months after dose escalation Patients are interviewed twice a month and then monthly thereafter to measure blood pressure. Each side Symptom severity scores per interview are based on the patient's experience 24 hours prior to the interview.Example 2   In addition to treating patients with interstitial cystitis, patients with urethral syndrome are referred to US Pat. Treatment with a compound of formula I using the treatment protocol and titration test described in No. 59 Was. Similar to the data in Example 1, a clear reaction of the compounds of the invention in this limited test was observed. In response, both urethral syndrome and interstitial cystitis probably have reflex sympathetic nervous system Hypothesis that trophy changes are part of the same disease spectrum I support.   The preferred embodiments of the present invention have been described. Obviously, after reading this specification, Modifications and changes will occur in understanding. Such modifications and changes shall not It is to be understood that as long as they are within the scope and equivalents thereof, they are included in the present invention. Is done.   Compounds used in the method of the present invention can be administered directly without formulation Usually, however, the compound is combined with a pharmaceutically acceptable excipient and at least one active agent. It is administered in the form of a pharmaceutical composition containing the sexual component. The composition may be orally, rectally, transdermally, dermally It can be administered by a variety of routes, including, below, intravenously, intramuscularly and intranasally. Departure The various compounds used in the above methods are effective as injectable and oral compositions. You. Such compositions are prepared in a manner well known in the pharmaceutical art and comprise at least one Active compounds (eg, REMINGTON'S PHARMACEUTICAL SCIENCES, (first 6th edition, 1980)).   In preparing the compositions for use in the present invention, the active ingredients are usually mixed with excipients. Or diluted with excipients or in the form of capsules, sachets, paper or other containers Included in a possible carrier. When using excipients as diluents, Is a solid, semi-solid material that acts as a vehicle, carrier or medium for the active ingredient. Or a liquid substance. That is, the composition comprises tablets, pills, powders, Agent, sachet, cachet, elixir, suspension, emulsion, Solutions, syrups, aerosols (as a solid or in a liquid medium), such as Ointments, soft and hard gelatin capsules containing up to 10% by weight of active compound , Suppositories, sterile injectable solutions, and sterile packaged powders.   In the preparation of the preparation, the active compound is comminuted and mixed with the appropriate ingredients before mixing with the other ingredients. May need to be child size. Where the active compound is substantially insoluble If so, the compound is usually ground to a particle size of less than 200 mesh. Activation If the product is substantially water-soluble, the particle size will typically be substantially uniform throughout the formulation. Is adjusted by grinding (eg, about 40 mesh) .   Examples of suitable excipients are lactose, glucose, sucrose, sorbitol, mannitol , Starch, gum acacia, calcium phosphate, alginate, tragacanth, Gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cell Includes loin, water, syrup, and methylcellulose. In addition, the formulation Are lubricating and wetting agents such as talc, magnesium stearate, and mineral oil , Emulsifiers and suspending agents, such as methyl- and propylhydroxybenzoate. Such preservatives, sweeteners, and flavoring agents can be included. The composition of the present invention After administration to a patient using methods known in the art, the active ingredient is rapidly or sustainedly released. It can be formulated for release with or without delay.   The compositions may contain from about 0.05 to about 100 mg, more usually about 1 to about 100 mg, of active ingredient per dose. . It is preferred to formulate a unit dosage form containing from 0 to about 30 mg. The term `` unit throw The "dosage form" is a pre-determined, unit calculated for each unit to produce the desired therapeutic effect. Human subjects and other mammals containing a reduced amount of active substance with suitable pharmaceutical excipients A physically discrete unit suitable as a unit dosage for a product.   The active compound is generally effective over a wide dosage range. For example, one day Dosage ranges from about 0.01 to about 30 mg / kg body weight. In adult treatment Has a range of about 0.1 to about 15 mg / kg / day (single or divided dose). Preferred. However, the actual amount of compound administered will depend on the condition being treated, The chosen route of administration, the actual compound or compounds to be administered, the individual patient Physicians in light of relevant circumstances, including age, weight, and response, and the severity of the patient's symptoms The above dose ranges limit the scope of the invention in any way, as will be determined by the pharmacist. Not something. In some cases, dose levels below the lower end of the above range may be appropriate. However, in other cases larger doses can be used without any adverse side effects. (Although such large doses may initially be administered throughout the day) Divided into some smaller amounts).Formulation Example 1   A hard gelatin capsule containing the following ingredients is prepared.   component Amount (mg / capsule)   Active ingredient 30.0   Starch 305.0   Magnesium stearate 5.0   The above ingredients are mixed and 340 mg is filled into hard gelatin capsules.Formulation Example 2   A tablet is prepared using the following ingredients.   component Amount (mg / tablet)   Active ingredient 25.0   Cellulose, microcrystalline 200.0   Colloidal silicon dioxide 10.0   Stearic acid 5.0   The ingredients are mixed and compressed to form tablets weighing 240 mg.Formulation Example 3   A dry powder inhalation formulation containing the following ingredients is prepared.component weight% Active ingredient 5 Lactose 95   The active mixture is mixed with lactose and the mixture is added to a dry powder inhaler.Formulation Example 4   A tablet containing 30 mg of the active ingredient is prepared as follows.  component Amount (mg / tablet)  Active ingredient 30.0  Starch 45.0  Microcrystalline cellulose 35.0  Polyvinylpyrrolidone 4.0  (As a 10% aqueous solution)  Sodium carboxymethyl starch 4.5  Magnesium stearate 0.5  Talc1.0   120mg   The active ingredients, starch and cellulose were no. 20 mesh U.S. S. Sieve Through and mix thoroughly. The polyvinylpyrrolidone solution is mixed with the resulting powder, This is 16 mesh U.S. S. Pass through sieve. 50 granules so produced ~ 60 ° C, 16 mesh U.S. S. Pass through sieve. Next, . 30 mesh U.S. S. Sodium carboxymethyl sed starch through sieve , Magnesium stearate, and talc are added to the granules, mixed, and then compressed. To obtain tablets weighing 120 mg each.Formulation Example 5   A capsule containing 40 mg of the active ingredient is prepared as follows.component Amount (mg / capsule) Active ingredient 40.0mg Starch 109.0mg Magnesium stearate 1.0mg Total amount 150.0mg   Mix active ingredients, cellulose, starch, and magnesium stearate , No. 20 mesh U.S. S. Through a sieve, then add 150 mg of hard gelatin Fill into capsules.Formulation Example 6   A suppository containing 25 mg of each active ingredient is prepared as follows.component amount Active ingredient 25mg Saturated fatty acid glyceride 2000mg (final amount)   The active ingredient was no. 60 mesh U.S. S. Sift through and minimize necessary Suspended in saturated fatty acid glyceride dissolved by heat. The mixture is then charged to a display capacity of 2. Pour into 0 g suppository mold and cool.Formulation Example 7   Produce a suspension containing 50 mg of active ingredient per 5.0 mL dose as follows .component amount Active ingredient 50.0mg Xanthan gum 4.0mg Sodium carboxymethyl cellulose (1 10.0%   Microcrystalline cellulose (89%) 1.75 g sucrose Sodium benzoate 10.0mg Flavors and colorings Purified water 5.0 mL (final volume)   The active ingredient, sucrose and xanthan gum were mixed together. 10 mesh U.S. S . Sieve, then pre-made microcrystalline cellulose and sodium Mix with an aqueous solution of carboxymethyl cellulose. Sodium benzoate, flavor And the color is diluted with some of the water and added with stirring. Next, a sufficient amount of water Was added to obtain the required amount.Formulation Example 8   A capsule containing 15 mg of the active ingredient is prepared as follows.component Amount (mg / capsule) Active ingredient 15.0mg 407.0mg starch Magnesium stearate 3.0mg 425.0mg in total   Mix active ingredients, cellulose, starch, and magnesium stearate , No. 20 mesh U.S. S. Pass 425 mg of hard gelatin Fill the cell.Formulation Example 9 An intravenous formulation could be manufactured as follows.component amount Active ingredient 250.0mg Isotonic saline 1000mLFormulation Example 10   A topical formulation could be manufactured as follows.component amount Active ingredient 1-10g 30g wax for emulsification Liquid paraffin 20g 100g white soft paraffin (final amount)   Heat until white soft paraffin dissolves. Liquid paraffin and emulsifying wax Mix and stir until dissolved. Add the active ingredient and continue stirring until dispersed. Next, the mixture is cooled until it becomes solid.Formulation Example 11   A sublingual or buccal tablet containing 10 mg of active ingredient could be prepared as follows: .component Amount / tablet Active ingredient 10.0mg Glycerol 210.5mg 143.0 mg of water 4.5 mg sodium citrate 26.5mg of polyvinyl alcohol Polyvinyl pyrrolidone15.5mg Total amount 410.0mg   Glycerol, water, sodium citrate, polyvinyl alcohol and polyvinyl alcohol Maintain nilpyrrolidone at a temperature of about 90 ° C and mix together with continuous stirring You. Once the polymer is in solution, cool the solution to about 50-55 ° C and slowly release the active ingredients. Mix separately. Pour the homogeneous mixture into a mold made of inert material and make it about 2-4 mm of drug-containing diffusion matrix is obtained. Next, cut this dispersion matrix, Form individual tablets of appropriate size.   Another preferred formulation for use in the method of the invention uses a transdermal delivery device ("patch"). I have. Using such transdermal patches, a controlled amount of a compound of the present invention can be administered continuously or Could be injected discontinuously. Of transdermal patches to deliver medicinal substances Construction and use are well known in the art (eg, US Pat. No. 5,023,23). No. 252 (issued on June 11, 1991), the contents of which are incorporated in this specification. Do). Such patches can deliver the drug substance continuously, pulsed, or as needed. Could be built to supply.   It is preferred or necessary to introduce the pharmaceutical composition directly or indirectly into the brain There will be many. Direct techniques usually bypass the blood-brain barrier Involves placing a drug delivery catheter within the host's ventricular system. Biological factors Such an implantable supply used to transport a child to a specific anatomical area of the body The system is disclosed in U.S. Pat. No. 5,011,472, issued Apr. 30, 1991, The contents of which are incorporated in this specification).   In general, preferred indirect techniques typically involve replacing hydrophilic drugs with lipophilic drugs or drugs. Latency of drugs by converting them to lodrugs (delay and latency) And formulating the composition to provide Latency is , In general, to make the drug more lipophilic and to be transported across the blood-brain barrier Hydroxy, carbonyl, sulfate and primary present in the drug to Achieved by blocking amine groups. Alternatively, supply of hydrophilic drugs Feeding is increased by intraarterial infusion of a hypertonic solution that can transiently open the blood-brain barrier Could be done.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme court ゛ (Reference) C07D 401/12 C07D 401/12 (81) Designated country EP (AT, BE, CH, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG), AP (GH, KE, LS, MW, SD, SZ, UG), UA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GE, GH, HU, IL, S, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO , RU, SD, SE, SG, SI, SK, TJ, TM, TR, TT, UA, UG, US, UZ, VN, YU (72) Inventor Saw, Carl Bee United States Draymore Way No. 109 [Summary Continued] Provides a method to include.

Claims (1)

[Claims] 1. A method for treating or preventing interstitial cystitis or urethral syndrome in a mammal, comprising administering to a mammal in need thereof an effective amount of a formula: Wherein R ¹ and R ² are independently selected from the group consisting of hydrogen, methyl, methoxy, chloro and trifluoromethyl (provided that only one of R ¹ and R ² can be hydrogen ), Y is A compound represented by the formula: NR ^a or CH-NR ^b R ^c (where R ^a , R ^b and R ^c are independently selected from the group consisting of hydrogen and C ₁ -C ₆ alkyl)] Or administering a pharmaceutically acceptable salt or solvate thereof. 2. (R) -3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) acetylamino] -2- [N- (2- (4-piperidin-1-yl) piperidin- The method according to claim 1, wherein 1-yl) acetyl) amino] propane or a pharmaceutically acceptable salt or solvate thereof is used. 3. (R) -3- (1H-indol-3-yl) -1- [N- (2-methoxybenzyl) acetylamino] -2- [N- (2- (4-piperidin-1-yl) piperidin- 3. The method according to claim 2, wherein 1-yl) acetyl) amino] propane dihydrochloride trihydrate is used. 4. The compound according to any one of claims 1 to 3, which is used for treating or preventing interstitial cystitis or urethral syndrome. 5. A pharmaceutical preparation comprising the compound according to any one of claims 1 to 3 as an active ingredient, which is used for treating or preventing interstitial cystitis or urethral syndrome. 6. Use of a compound according to any of claims 1 to 3 for the manufacture of a medicament for use in treating or preventing interstitial cystitis or urethral syndrome.