Amide derivatives and 5-lipoxygenase inhibitors containing the same as an active ingredient

Abstract: Provided are amide derivatives having the general formula (I): wherein R¹ represents hydrogen atom or an alkyl group, R² represents hydrogen atom, an alkyl group or toluoyloxy group, or R¹ and R² when taken together represent an alkylene group, R³ represents hydrogen atom or an alkoxy group, R² and R³ both being hydrogen atom when R¹ represents hydrogen atom, and n is an integer of 1 or 2; Y represents an amino group having the formula wherein X⁶ represents hydrogen atom, a halogen atom or an alkoxy group. The above-mentioned compounds have been demonstrated to possess 5-lipoxygenase-inhibiting activities. These compounds can inhibit the production of leuco­ trienes such as LTC₄ and LTD₄ which are allergy-inducing factors by inhibiting the activity of 5-lipoxygenase. Accordingly, the amide derivatives can be used as 5-lipoxygenase inhibitors effective for allergic asthma, allergic rhinitis and the like.

Description: Background of the Invention 1. Field of the Invention The present invention relates to novel amide derivatives and 5-lipoxygenase inhibitors containing the same as an active ingredient. The amide derivatives provided by the invention possess activities inhibiting 5-lipoxygenase which is an enzyme. Leucotrienes such as leucotriene C₄(LTC₄) and leucotriene D₄(LTD₄) which are allergy-inducing factors are biosynthesized in vivo from arachidonic acid by the action of 5-lipoxygenase. Therefore, the amide derivatives of the invention, which possess 5-lipoxygenase-inhibiting activities, will inhibit biosynthesis of the above-mentioned allergy-inducing factors thereby being useful as antiallergic agents which can effectively be used for allergic asthma or allergic rhinitis . 2. Description of the Prior Art It has recently been found that action of 5-lipoxygenase on arachidonic acid results in formation of leucotrienes which are allergy-inducing factors (Science vol. 220, page 568, 1983, published by the American Association for the Advancement of Science). As described above, leucotrienes such as LTC₄ and LTD₄ which are 5-lipoxygenase products from arachidonic acid are important factors participating in onset of diseases such as allergic asthma and allergic rhinitis, and therefore, development of drugs that are of 5-lipoxygenase-inactivating activities and capable of inhibiting its action is strongly desired. Summary of the Invention As a results of studies on the synthesis of amide derivatives and their 5-lipoxygenase-inhibiting activities, we have found that the amide derivatives according to the present invention possess high 5-lipoxygenase-inhibiting activities. The invention has been completed based upon such finding. It is an object of the invention to provide novel amide derivatives and 5-lipoxygenase inhibitors containing the same as an active ingredient. Detailed Description of the Invention According to the present invention, there are provided amide derivatives having the general formula: In the formula (I), R¹ represents hydrogen atom or an alkyl group with 1 to 4 carbon atom(s), R² represents hydrogen atom, an alkyl group with 1 to 4 carbon atom(s) or toluoyloxy group, or R¹ and R² when taken together represent an alkylene group with 1 to 3 carbon atom(s), R³ represents hydrogen atom or an alkoxy group with 1 to 4 carbon atom(s), R² and R³ both being hydrogen atom when R¹ represents hydrogen atom, and n is an integer of 1 or 2 and Y represents an amino group having the formula wherein X⁶ represents hydrogen atom, a halogen atom or an alkoxy group with 1 to 4 carbon atoms, with the proviso that if n is 1, at least one of R¹ and R² must be hydrogen. Preferred are the amide derivatives selected from: N-benzhydryl-N'-{(3,4-dihydroxyphenyl)-2-propenoyl}-piperazine; 1-benzhydryl-4-{5-(3-methoxy-4-hydroxyphenyl)-2,4-petadienoyl}-piperazine or 1-benzyhdryl-4-{5-(3,4-dihydroxyphenyl)-2,4-pentadienoyl}-piperazine. Further according to the present invention, a pharmaceutical composition containing an amide derivative of having the general formula (I): wherein R¹ represents a hydrogen atom or an alkyl group with 1 to 4 carbon atoms, R² represents a hydrogen atom , an alkyl group with 1 to 4 carbon atoms or a toluoyloxy group, or R¹ and R² when taken together represent an alkylene group with 1 to 3 carbon atoms, R³ represents a hydrogen atom or an alkoxy group with 1 to 4 carbon atoms, R² and R³ both being a hydrogen atom when R¹ represents a hydrogen atom, and n is an integer of 1 or 2; Y represents an amino group having the formula wherein X⁶ represents a hydrogen atom, a halogen atom or an alkoxy group with 1 to 4 carbon atoms with the proviso that if n is 1, at least one of R¹ and R² must be hydrogen. and a pharmaceutically acceptable carrier is provided. Further, there is provided the use of an amide derivative selected from: N-benzhydryl-N'-{3,4-dihydroxyphenyl)-2-propenoyl}-piperazine; 1-benzhydryl-4-{5-(3-methoxy-4-hydroxyphenyl)-2,4-petadienoyl}-piperazine or 1-benzyhdryl-4-{5-(3,4-dihydroxyphenyl)-2,4-pentadienoyl}-piperazine. In the definitions of the above formula, the alkyl group containing 1 - 4 carbon atoms is for example methyl, ethyl, n-propyl, isopropyl, n-butyl or isobutyl. The alkoxy group containing 1 - 4 carbon atoms is for example methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy or isobutoxy. The alkylene group containing 1 - 3 carbon atoms is for example methylene, ethylene, trimethylene or 1-methylethylene. The halogen atom preferably means chlorine, bromine or jodine. The amide derivatives having the above formula (I) are novel compounds which are produced, for example, by reacting a carboxylic acid having the formula wherein R¹, R², R³ and n are as defined above or a reactive derivative thereof such as a compound having the formula wherein R¹, R², R³ and n are as defined above with an amine having the formula H - Y (IV) wherein Y is as defined above and, if required, subjecting the reaction product to a deprotection reaction. The amide derivatives of the invention are useful as 5-lipoxygenase inhibitors or antiallergic agents. The dosage will generally be in the range from 30 to 200 mg and preferably from 50 to 600 mg per day in adults, although it is variable depending upon of the patient. It may be divided into one to three doses per day as required depending upon conditions of the patient. The administration may be by any suitable route. Intravenous injection is applicable, though oral administration is particularly desirable. The compounds of the invention may be blended with pharmaceutical carriers or excipients and formulated into a variety of forms such as tablets, sugar-coated tablets, powders, capsules, granules, suspension, emulsion or injectable solution. As examples of the carrier or excipient are mentioned calcium carbonate, starch, glucose, lactose, dextrin, arginic acid, mannitol, talc and magnesium stearate . Examples and Test Example will be given below to describe the invention in more details. Example 1 To a solution of 198 mg (0.79 mmol) of benzhydryl-piperazine in 5 ml of tetrahydrofuran was added in an argon atmosphere a solution of 205 mg (0.45 mmol) of N-3-{3,4-di(β-methoxyethoxymethoxy)phenyl}-2-propenoylthiazolidine-2-thione in 5 ml of tetrahydrofuran. The mixture was reacted at room temperature for 10 min. To the reaction mixture was added 20 ml of 2 N -aqueous solution of sodium hydroxide, and the mixture was extracted with ethyl acetate. The organic layer was concentrated under reduced pressure, and the residue thus obtained was subjected to silica gel column chromatography. There was obtained from a fraction eluted with chloroform - methanol (100 : 1) 241 mg (0.41 mmol) of N-benzhydryl-N'-[3-{3,4-di(β-methoxyethoxymethoxy)-phenyl}-2-propenoyl]piperazine. To a solution of 170 mg (0.29 mmol) of the amide compound prepared above in 5 ml of methanol was added in an argon atmosphere 52 mg (0.30 mmol) of p -toluene-sulfonic acid. The mixture was heated under reflux for 3 hours. To the reaction mixture was added 20 ml of saturated aqueous solution of sodium hydrogen carbonate, and athe mixture was extracted with ethyl acetate. The organic layer was concentrated under reduced pressure, and the residue thus obtained was recrystallized from methanol to give 95 mg (0.23 mmol) of N-benzhydryl-N'-{(3,4-dihydroxyphenyl)-2-propenoyl}-piperazine. Spectrophotometric data of the product support the structure shown below. 3475, 3125, 1645, 1600, 1570, 1530. ¹H-NMR (deutero pyridine) δ : 2.33 (4H, m), 3.70 (4H, m), 4.28 (1H, s), 6.90 - 7.57 (14H, m), 8.02 (1H, d, J=16Hz) Example 2 To a solution of 15.2 g (0.10 mol) of vanillin in 200 ml of dichloroethane were added in an argon atmosphere 13 ml (0.114 mol) of β-methoxyethoxymethyl chloride and 21 ml (0.121 mol) of diisopropylethylamine. The mixture was heated under reflux for 3 hours, followed by addition of water. The mixture was extracted with chloroform, and the organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The residue thus obtained was subjected to silica gel column chromatography. There was obtained from a fraction fluted with benzene - ethyl acetate (10 : 1) 22 g (0.092 mol) of 3-methoxy-4-β-methoxyethoxymethoxybenzaldehyde. In 100 ml of dry tetrahydrofuran was dissolved 1.133 g (47 mmol) of the aldehyde prepared above. Separately, to 150 ml of dry tetrahydrofuran were added in an argon atmosphere 2 g (50 mmol) of 60 % sodium hydride and then 11 ml (50 mmol) of triethyl 4-phosphonocrotonate. The mixture was reacted at room temperature for 30 min. To the reaction solution was added the tetrahydrofuran solution of the aldehyde prepared above. The mixture was reacted at room temperature for 3 hours, followed by addition of saturated aqueous solution of ammonium chloride. The resulting mixture was extracted with ethyl acetate. The organic layer was concentrated under reduced pressure, and the residue thus obtained was subjected to silica gel column chromatography. There was obtained from a fraction eluted with benzene - acetone (9 : 1) 9.0 g (27 mmol) of ethyl 5-(3-methoxy-4-β-methoxyethoxy-methoxyphenyl)-2,4-pentadienate. Then, to a solution of 8.8 g (26 mmol) of the ester prepared above in 100 ml of methanol were added in an argon atmosphere 25 ml of water and 10.4 g (260 mmol) of sodium hydroxide. The mixture was reacted at room temperature for 1 hour. 6 N -hydrochloric acid was added to the reaction solution to adjust the pH to 4, followed by addition of 200 ml of water. The resulting mixture was extracted with ethyl acetate, and the organic layer was concentrated under reduced pressure to give 7.6 g (24.7 mmol) of 5-(3-methoxy-4-β-methoxyethoxymethoxyphenyl)-2,4-pentadienic acid. Then, to a solution of 2.08 g (6.74 mmol) of the carboxylic acid prepared above in dry dichloroethane (40 ml) were added 996 mg (8.36 mmol) of 2-mercapto-thiazo-line, 109 mg (0.892 mmol) of dimethylaminopyridine and 1.82 g (8.83 mmol) of N,N'-dicyclohexylcarbodiimide. The mixture was reacted in an argon atmosphere at room temperature for 14 hours. Precipitates then formed were removed by filtration, and the filtrate was concentrated under reduced pressure. The residue thus obtained was subjected to silica gel column chromatography. There was obtained from a fraction eluted with chloroform 2.62 g (6.40 mmol) of N-5-(3-methoxy-4-β-methoxyethoxymethoxyphenyl)-2,4-pentadienoyl-2-thiothiazolidine. To a solution of 542 mg (1.32 mmol) of the amide compound prepared above in 10 ml of tetrahydrofuran was added in an argon atmosphere a solution of 603 mg (2.39 mmol) of benzhydrylpiperazine in tetrahydrofuran (10 ml). The mixture was reacted at room temperature for 10 min., followed by addition of 40 ml of 2 N -aqueous solution of sodium hydroxide. The resulting mixture was extracted with ethyl acetate, and the organic layer was concentrated under reduced pressure. The residue thus obtained was subjected to silica gel column chromatography. There was obtained from a fraction eluted with chloroform - methanol (100 : 1) 616 mg (1.14 mmol) of 1-benzhydryl-4-{5-(3-methoxy-4-β-methoxyethoxymethoxyphenyl)-2,4-pentadienoyl}piperazine. To a solution of 120 mg (0.221 mmol) of the amide compound prepared above was added in an argon atmosphere 46 mg (0.242 mmol) of p -toluenesulfonic acid hydrate. The mixture was refluxed for 1 hour. The reaction solution was concentrated under reduced pressure, followed by addition of water. The resulting mixture was adjusted with aqueous solution of sodium carbonate to a pH of 10, followed by extraction with ethyl acetate. The organic layer was concentrated under reduced pressure, and the residue thus obtained was subjected to silica gel thin layer chromatography. Development with chloroform - methanol (40 : 1) yielded 86 mg (0.189 mmol) of 1-benzhydryl-4-{5-(3-methoxy-4-hydroxyphenyl)-2,4-pentadienoyl}piperazine. Spectrophotometric data of the product support the structure shown below. 3450, 1640, 1585. ¹H-NMR (deutero-chloroform) δ : 2.40 (4H, bs), 3.63 (4H, bs), 3.83 (3H, s), 4.23 (1H, s), 6.35 (1H, d, J=15Hz), 6.70 - 7.25 (16H, m). Example 3 To a solution of 470 mg (1.87 mmol) of benzhydrylpiperidine in 5 ml of tetrahydrofuran was added in an argon atmosphere a solution of 396 mg (0.82 mmol) of N-5-{3,4-di(β-methoxyethoxymethoxy)phenyl-2,4-pentadienoyl-2-thio-thiazolidine in 5 ml of tetrahydrofuran. The mixture was reacted at room temperature for 10 min. To the reaction solution was added 20 ml of 2 N -aqueous solution of sodium hydroxide, and the mixture was extracted with ethyl acetate. The organic jlayer was concentrated under reduced pressure, and the residue thus obtained was subjected to silica gel column chromatography. There was obtained from a fraction eluted with chloroform-methanol (100 : 1) 497 mg (0.81 mmol) of 1-benzhydryl-4- [5-{3,4-di(β-methoxyethoxy-methoxy)phenyl}-2,4-pentadienoyl]piperazine. To a solution of 472 mg (0.77 mmol) of the amide compound prepared-above in 5 ml of methanol was added in an argon atmosphere 138 mg (0.80 mmol) of p -toluenesulfonic acid. The mixture was heated under reflux for 3 hours. To the reaction solution was added 20 ml of saturated aqueous solution of sodium hydrogen carbonate, and the mixture was extracted with ethyl acetate. The organic jlayer was concentrated under reduced pressure, and the residue thus obtained was recrystallized from methanol to give 270 mg (0.61 mmol) of 1-benzhydryl-4-{5-(3,4-dihydroxyphenyl)-2,4-pentadienoyl}piperazine. Spectrophotometric data of the product support the structure shown below. 3475, 3100, 1635, 1615, 1595, 1565. ¹H-NMR (deutero pyridine) δ : 2.33 (4H, m), 3.70 (4H, m), 4.28 (1H, s), 6.62 (1H, d, J=16Hz), 6.90 - 8.10 (16H, m). Test Example 5-Lipoxygenase-inhibiting Activity Mouse mastcytoma cells P-815 are diluted in a culture solution, containing 90 % Eagle Basic Medium (manufactured by Gibco Laboratories) to 5 x 10⁴ cells/ml. The diluted solution is cultivated with shaking in air at 37 °C for 48 hours. Then, the culture solution is cooled with ice and centrifuged to collect the cells. The cells are suspended in a, phosphate buffer solution at pH 7.4 at a concentration of 2 x 10⁷ cells/ml. The suspension is treated in a ultrasonic cell disintegrater and then centrifuged at 10,000 r.p.m. for 10 min. The supernatant is used as a 5-lipogenase enzyme solution. In test tubes are placed 20 µl of radiolabelled arachidonic acid (10 Ci/ml), indomethacin (2 x 10⁻⁸ mol) and an amide derivative to be tested, respectively. To each of the test tubes are added 0.45 ml of a phosphate buffer solution, 0.45 ml of the above-prepared suspension, 0.45 ml of the above-prepared enzyme solution and 0.1 ml of 8 mM CaCl₂ (calcium chloride). The mixture is reacted at 37 °C for 5 min. After cooled with ice, 60 µl of 1 N -HCl (hydrochloric acid is added to the reaction mixture, which is then extracted with 8 ml of ethyl acetate. The extract is concentrated, and the concentrate is developed by spotting on a silica gel thin layer plate (Merck 60F ₂₅₄). Measurement of the inhibitory activity is made by collecting fractions corresponding to lipoxygenase products 5-HETE (5(s)-hydroxy-6,8,11,14-eicosatetraenic acid) and LTB₄ (leucotriene B₄) and determining the radioactivity by a liquid scintillation counter The lipoxygenase inhibiting activity is determined by decrease in the amount of the above-mentioned 5-lipoxygenase products. There have been found remarkeable 5-lipoxygenase-inhibiting activities as shown in Table I below. It has also been found that the amide derivatives of the invention other than those shown in Table I possess 5-lipoxygenase-inhibiting activities. 50 % inhibitory concentration or 30 % inhibitory concentration as indicated in the table means the concentration of an amide derivative required for reducing the amount of the above-mentioned 5-lipoxygenase products (5-HETE and LTB₄) to 50 % or 30 % of the amount when the amount of 5-HETE and LTB₄ produced in the absenced of the amide derivative is 100 %. Acute Toxicity An acute toxicity test was run in ICR male mice (5-week old) by oral administration. LD₅₀'s with all of the compounds of the invention were 100 mg/kg body weight or higher, by which the high safety as compared with the efficacy was demonstrated. 5-Lipoxygenase-inhibiting activities Examples No. 50% Inhibitory concentration IC₅₀ (mmol) 1 1 x 10⁻⁷ 2 9 x 10⁻⁸ 3 4 x 10⁻⁸

Claims: An amide derivative having the general formula (I): wherein R¹ represents a hydrogen atom or an alkyl group with 1 to 4 carbon atoms, R² represents a hydrogen atom , an alkyl group with 1 to 4 carbon atoms or a toluoyloxy group, or R¹ and R² when taken together represent an alkylene group with 1 to 3 carbon atoms, R³ represents a hydrogen atom or an alkoxy group with 1 to 4 carbon atoms, R² and R³ both being a hydrogen atom when R¹ represents a hydrogen atom, and n is an integer of 1 or 2; Y represents an amino group having the formula wherein X⁶ represents a hydrogen atom, a halogen atom or an alkoxy group with 1 to 4 carbon atoms with the proviso that if n is 1, at least one of R¹ and R² must be hydrogen. An amide derivative of claim 1 selected from: N-benzhydryl-N'-{(3,4-dihydroxyphenyl)-2-propenoyl}-piperazine; 1-benzhydryl-4-{5-(3-methoxy-4-hydroxyphenyl)-2,4-petadienoyl}-piperazine or 1-benzhydryl-4-{5-(3,4-dihydroxyphenyl)-2,4-pentadienoyl}-piperazine. A pharmaceutical composition containing an amide derivative of claims 1 or 2 and a pharmaceutically acceptable carrier. Use of an amide derivative having the general formula (I): wherein R¹ represents a hydrogen atom or an alkyl group with 1 to 4 carbon atoms, R² represents a hydrogen atom , an alkyl group with 1 to 4 carbon atoms or a toluoyloxy group, or R¹ and R² when taken together represent an alkylene group with 1 to 3 carbon atoms, R³ represents a hydrogen atom or an alkoxy group with 1 to 4 carbon atoms, R² and R³ both being a hydrogen atom when R¹ represents a hydrogen atom, and n is an integer of 1 or 2; Y represents an amino group having the formula wherein X⁶ represents a hydrogen atom, a halogen atom or an alkoxy group with 1 to 4 carbon atoms for the manufacture of a 5-lipoxygenase inhibiting pharmaceutical composition . Use of an amide derivative selected from: N-benzhydryl-N'-{(3,4-dihydroxyphenyl)-2-propenoyl}-piperazine; 1-benzhydryl-4-{5-(3-methoxy-4-hydroxyphenyl)-2,4-petadienoyl}-piperazine or 1-benzhydryl-4-{5-(3,4-dihydroxyphenyl)-2,4-pentadienoyl}-piperazine. A process for the preparation of a compound according to anyone of claims 1 or 2 by reacting a carboxylic acid having the formula (II) wherein R¹, R², R³ and n are as defined in claim 1, or a reactive derivative thereof with an amine of the formula (IV) H - Y (IV) wherein Y is as defined in claim 1 and, if required, subjecting the reaction product to a deprotection reaction.

Inventors: ARAI JUN-ICHIRO, ICHIKAWA SHUJI, MUROTA SEIITSU, TAKAI MAKOTO, WAKABAYASHI TOSHIO, ARAI, JUN-ICHIRO, ICHIKAWA, SHUJI, MUROTA, SEIITSU, TAKAI, MAKOTO, WAKABAYASHI, TOSHIO
Applicants: TERUMO CORP, TERUMO KABUSHIKI KAISHA TRADING AS TERUMO CORPORATION
Application year: 1985
Main classification: C07D295
IPC codes: C07D295