WO1996034011A1

WO1996034011A1 - Purifying methods

Info

Publication number: WO1996034011A1
Application number: PCT/JP1996/001141
Authority: WO
Inventors: Kazuya Nishimura; Michinori Ozawa; Akira Kagemoto
Original assignee: Dainippon Pharmaceutical Co., Ltd.
Priority date: 1995-04-27
Filing date: 1996-04-25
Publication date: 1996-10-31
Also published as: AU5514496A

Abstract

A method of purifying crude (2S, 3aS, 7aS)-1-(N2-nicotinoyl-L-lysyl-η-D-glutamyl)octahydro-1H-indole-2-carboxylic acid (DU-1777) by cristallizing the same from methanol; and another method of purifying crude DU-1777 crystals by recrystallizing the same from a solvent mixture comprising a lower alkanol and at least one solvent selected among water, ethylene glycol and propylene glycol. These methods are industrially advantageous, because they can provide high-purity DU-1777 applicable as a medicine through simple operations.

Description

Akira Tsumugi Refining method Technical field

The present invention is useful as a cardiovascular disease treating agent (2S, 3aS, 7a S) - 1 - (N 2 - Nikochino I le one L over Li Giroux 7 - D - glutamicum Le) O click evening arsenide The present invention relates to an industrially advantageous purification method by crystallization and recrystallization of draw 1 H—indole 2 —carboxylic acid (hereinafter sometimes referred to as “DU—17777”). Background art

DU-17777 is a compound represented by the following formula. US Pat. No. 4,826,814 (JP-B 5-37998) discloses that this compound is a potent angiotensin. It is disclosed that it exhibits a converting enzyme inhibitory effect and a strong and sustained antihypertensive effect.

In Example 15 of the above-mentioned U.S. Patent, three types of production examples of DU-17777 are described. In any of the production examples, DU-17777 is a reversed-phase column chromatograph. Purified by a combination of fy and lyophilization and is obtained in amorphous form. That is, the reaction solution containing DU-177 7 was appropriately treated according to the reaction conditions, and then the mobile phase was changed to acetonitrile / CHP20P (0% → 60% gradient of water mixed solvent) (High-porous polystyrene resin: Applied to column chromatography of 75 to 150 〃π. Fraction containing DU-177 7 Is concentrated in vacuo, then dissolved in water and freeze-dried to obtain amorphous DU-177 7. In the case of purification of a small amount of DU-177 7 It is possible to continuously change the tritium content as described above, and it is possible to easily obtain pure DU-17777. It is difficult to continuously change the amount of acetonitrile in the mobile phase in the production of DU-177 7 in Japan, usually 10%, 20%, 30%. Therefore, high-purity DU-177 7 can be obtained only by the operation of reversed-phase column chromatography. If this is not possible, or if DU-177 7 is crystallized, it may be possible to purify it by recrystallization.Therefore, the present inventors have conducted intensive studies on crystallization of DU-177 7 Under certain conditions, DU-177 7 is crystallized, and when the resulting crude crystals of DU-177 7 are recrystallized from a specific solvent, the purity of DU-177 7 increases. The present invention has been completed and the present invention has been completed.

The present invention relates to a crude (2S, 3aS, 7aS)-1-(N ² nicotinol-1 L-lysyl-1-D-daltamyl) emissions de one Lou 2 - a force carboxylic acid menu evening with Bruno Lumpur characterized the crystallization child (2S, 3aS, 7aS) - 1 one (N ² - nicotinonitrile I le - L - lysyl - § one D- guru evening mil) O click evening purification method arsenide mud one 1 H- Lee emissions Doll one 2 _ carbonitrile phosphate and, (2) (2S, 3aS.7aS ) - 1 - (N 2 - Nicotinoy Joichi L—Rigiru 7'—D—Glutami 1H—Indone 2—Crystalline ruboric acid in water, ethylenic alcohol, and propylene One or more solvents selected from glycols and lower alkanols Wherein the recrystallization child from a mixed solvent of Le (2S.3aS, 7aS) - 1 one (N ² one Nikochino Iru L -? Li Giroux ■ - D-Group evening mil) O click evening arsenide draw 1 It is intended to provide a method for purifying H-indole-2-carboxylic acid.

“Crude D U— 1 7 7 7” refers to amorphous D U with a purity of about 85% or more.

-Means 1 7 7 7 The “crude crystal of DU-1777” means DU-17777 purified by crystallizing crude DU-1777 using methanol. The term "lower alkanol" refers to those having 1 to 3 carbon atoms, for example, methanol, ethanol, and isopropyl alcohol, and particularly preferred is methanol. "Crude DU" — 1 7 7 7 ”is acquired as follows, for example.

Reference Example 1 (1) to (4) or Reference Example 2 which is a method based on the method described in Example 15 of U.S. Pat. No. 4,826,814 (Japanese Patent Publication No. 5-37998). It was obtained according to the method described in (1) ~ (5), (2S, 3aS, 7aS - 1 one (N ² - Nikochino Iru N ⁶ - Application Benefits Furuoroasechiru one L- Li di le ten ¹ Echiru 7 "- D —Glutamyl) o-Hydro-1 H-indole-12 —Carboxylic acid or (2S, 3aS, 7aS) — 1- (N ⁶ —tert-butynecarbonyl—N ² — Nicotinol-L—Risil ——— D—Glugyl mil 1-octahydro-1-H—indole-1—Carbonic acid is prepared in a conventional manner, for example, Reference Example 1 (5) or Reference Example 2 below. Deprotect with alkali hydroxide or hydrochloric acid, respectively, according to the method described in (6) Next, concentrate the reaction solution under reduced pressure under a neutral or acidic condition, and concentrate the concentrated solution of DU-177 7 After that, the pH is adjusted to about 2 to about 5 with concentrated hydrochloric acid.The aqueous solution of DU-17777 obtained here is used as a filler for polystyrene-vinyl alcohol. For example, the solution is subjected to reverse-phase column chromatography using HP-20, SP-825 or SP-207 (both manufactured by Mitsubishi Kasei), and desalination treatment is performed by first flowing zion water. Next, if necessary, elution is carried out with 10% acetonitrile water to remove impurities, and then elution is carried out with 20 to 30% acetonitrile / water. A fraction containing 777 was obtained, and this was concentrated under reduced pressure to obtain “crude DU-1 777”.

Crude DU-17777 is usually obtained as a syrup-like substance, and about 6.5 to about 8 times (weight ratio) of methanol is added to the crude DU-177-77. After heating to about 60 ° C to dissolve, gradually cool it while stirring, and cool to about 3 ° C if necessary. “D U-177 7 7 coarse crystals” precipitate out and are collected by filtration. The purity of the crude crystals of D U-17777 varies slightly from lot to lot, but is usually about 97.0 to about 99.0%.

In order to purify the crude crystal of DU-17777 by recrystallization, first, the crude crystal is dissolved by heating in one or more solvents selected from water, ethylene glycol and propylene glycol. . The amount of the solvent is usually about 0.8 to about 2.0 times the weight of the crude crystal (weight ratio), and the heating temperature is usually about 45 ° C to about 65 ° C. C is appropriate. Then, about 7 to about 11 times (weight ratio) of low-grade AlNol is added to the crude crystal, and the mixture is again heated to about 45 ° C to about 65 ° C to dissolve. Allow to cool. High purity DU-177 7 7 crystals precipitate out and are collected by filtration. The optimum combination of the recrystallization solvent is a combination of water and methanol (volume ratio 1: about 8 to 10). In this case, DU-with a purity of about 99.5% or more is used. 1 7 7 7 is obtained. Even if the amount of 1Z5 to 1Z10 in the medium is replaced with acetonitrile, acetate or tetrahydrofuran during recrystallization, high-purity DU-17 7 7 can be obtained. If DU1777 having a purity of less than 99.5% is obtained, one or more solvents selected from water, ethylene glycol and propylene glycol, and a lower alcohol may be used again. It is desirable to carry out the recrystallization operation from a mixed solvent of toluene.

Due to the nature of the molecule, DU-17777 easily retains water of crystallization or crystallization solvent, and is incorporated into the molecule in any recrystallization solvent and dried. Therefore, removal is relatively difficult. In the case of pharmaceuticals, it is desirable to limit the content of methanol to less than 0.1% from the viewpoint of safety, so replace crystalline methanol with pharmaceutically acceptable ethanol. Is preferred. In order to carry out substitution with ethanol, the DU-177 7 crystal is heated and dissolved in about 0.8 to about 2.0 times (by weight) deionized water, and then DU-177 7 Add about 9 to about 11 times (weight ratio) of ethanol, and heat and dissolve again, then allow to cool. The precipitated crystals are collected by filtration and blow-dried at about 40 ° C to about 90 ° C to obtain DU-1777, which retains the crystal ethanol, which can be directly used as pharmaceuticals. It is applicable.

Removal of the crystallization solvent is carried out by dissolving purified DU-177 7 in deionized water, concentrating under reduced pressure if necessary, and then freeze-drying or spray-drying, whereby it can be applied as a pharmaceutical. DU-1 7 7 7 can be obtained. BEST MODE FOR CARRYING OUT THE INVENTION

Reference Examples and Examples will be given below to describe the present invention more specifically, but the present invention is not limited to these Examples. The compounds were identified by elemental analysis, mass spectra, IR spectra, and NMR spectra.

In the reference examples and examples, high-performance liquid chromatography (HPLC) was performed under the following conditions.

Column: YMOPACK A-312 0DS-A (Emmsie; 6 ø x 150 mm) Mobile phase: 0.2% trifluorofluoroacetic acid aqueous solution Zacetonitrile (80:20) Flow rate: 1 mlZ min

Temperature: 40 ° C

Detection: UV215 nm Reference example 1

Crude (2S, 3aS, 7aS) - 1 one (N ² one Nikochino I Lumpur - L- Li Giroux 7 - D- guru evening mil) O click evening heat draw 1 H- Lee emissions Doll one 2 - Chikararu Bonn Acquisition of acid (DU—177 7 7):

(1) N ² — Benziloquinone 2 N ⁶ — Trifluoroacetyl-L-Li-D-D-Duar Add 82.2 g of ethyl ethyl phthalate to 820 mL of medium and add at 40-50 ° C It was dissolved by heating. Under ice-cooling, 8.2 g of 5% palladium-carbon was added, and the mixture was catalytically reduced at 25 ° C for 1 hour in a stream of hydrogen. After the reaction mixture was filtered, the filtrate was concentrated under reduced pressure to obtain 59.8 g of N ⁶ -trifluoroacetyl-L-lysyl-D-glucaminate.

(2) Dissolve 40 g of the above-mentioned ziroxyl-functional lyponyl derivative in 400 ml of a mixture of tetrahydrofuran Z water (1: 1), and dicyclohexylamine at 25 ° C 18. lg Was added, and the mixture was stirred at the same temperature for 15 minutes.Next, N- (nicotinyloxy) succinic acid imidide 22.Og was added under ice cooling, and the mixture was stirred for 15 minutes, and then stirred at 25 ° C for 5 hours. After distilling off the tetrahydrofuran under reduced pressure, a 10% aqueous solution of sulfuric acid was added at 10 to 20 ° C to adjust the pH to 3, and the mixture was stirred for 1 hour at the same temperature. (PH 3), then washed with water, dried at 90 ° C, and dried at 90 ° C. N ² -Nicotinol-N ⁶ -Trifluoroacetyl-L-Lysyl-D- g.

(3) To a 390 nil suspension of acetonitrile of 39.3 g of the above nicotinol body was added 7.9 g of diisopropylamine at 25 ° C, followed by heating to 60 ° C and stirring for 1 hour. After cooling on ice, the crystals were collected by filtration, washed with acetonitrile, and dried at 60 ° C. 46.6 g of the crude crystals were suspended in a mixture of 22 ml of acetonitrile and 440 ml of acetone. The mixture was refluxed for one hour. After ice-cooling, and the precipitated crystals were collected by filtration, washed with acetone tons, and dried at 60 ° C, N ² - Nikochino I le - N ^e one Application Benefits off儿Oroasechi几one L one lysyl one D — 34.5 g of ethyl glutamate diisopropylamine salt was obtained. Melting point 1 85 to 1 88 ° C

(4) 34.5 g of the above dicotinoyl compound 'diisopropylamine salt and 13.3 g of (2S.3aS, 7aS one-year-old hydro- 1H-indole-12-carboxylate' hydrochloride) To 345 ml of methylene chloride, stirred under ice-cooling for 15 minutes, 10.9 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was added, and the mixture was stirred at the same temperature for 15 minutes. The mixture was further stirred for 4 hours at 25 ° C. The solvent was distilled off under reduced pressure, the residue was dissolved in ethyl acetate, and washed with a 10% aqueous solution of citric acid, a saturated aqueous solution of sodium bicarbonate, and a saturated saline solution in this order. , and dried over anhydrous sulfate Na Application Benefits um solvent was distilled off under reduced pressure (2S, 3aS.7aS) -. 1 - (N 2 one Nikochino I Lou N ⁶ - Application Benefits Furuoroa Sechiru L over Li Giroux 〇 ¹ Echiru § — D— Gluyi mil) Okuyi hydr 1 H—indole 1-2—Ethyl rubonate 39.0 g was obtained.

(5) Add 11.3 g of the above product to 22 ml of tetrahydrofuran, dissolve by heating at 60 ° C, cool on ice, and cool at 30 ° C or below 5N sodium hydroxide aqueous solution. 13.2 ml was added, followed by stirring at 25 ° C. for 8 hours. After adjusting the pH of the reaction solution to 7 with concentrated hydrochloric acid, tetrahydrofuran was distilled off under reduced pressure at 60 ° C or lower. The residual solution was adjusted to pH 3 with concentrated hydrochloric acid, applied to column chromatography using SP-825 (manufactured by Mitsubishi Kasei; polystyrene-styrene-based synthetic adsorbent), and deionized water was added. Desalting was performed by flowing. Next, after eluting with 10% aqueous acetonitrile to remove impurities, the desired fraction eluted with 20% aqueous acetonitrile Z was collected, reduced in E, concentrated and concentrated to obtain crude DU-. 1 7 7 7 was obtained. The purity determined by HPLC was 92.8%.

Reference example 2

Getting the crude DU-1 7 7 7:

(Π N ² benzene ziroxyka rubonil N ⁶ — tert-butene force liponyl L- lysyl −1 D—glucan ethyl 86.0 g added to 520 nil of methanol The mixture was heated and melted at 50 ° C. Under ice cooling, 8.6 g of 5% palladium-carbon was added, and the mixture was catalytically reduced at 25 ° C. for 1 hour in a stream of hydrogen. After the reaction solution was filtered, the filtrate was concentrated under reduced pressure to obtain 64.0 g of N ⁶ -tert-butoxycarbonyl-L-lysyl D-glucamin ethyl ester.

(2) Dissolve 64.0 g of the above debenzyloxycarbonyl compound in 640 ml of a mixture of tetrahydrofuran Z water (1: 1) and add 13.4 g of sodium bicarbonate at 25 ° C. The mixture was stirred at the same temperature for 30 minutes. Then N-

(Nicotinyloxy) 35.2 g of succinic acid imide was added, stirred for 20 minutes, and then stirred at 25 ° C for 6 hours. After distilling off tetrahydrofuran under reduced pressure, the pH was adjusted to 3 by adding a 10% aqueous solution of citric acid, and the oil was extracted with a black hole form. The organic layer is washed with a 10% aqueous solution of citric acid, water and saturated saline in this order, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and concentrated under reduced pressure to give N ⁶ -tert-butoxycarbonyl-N ² -nicotinol L-. 81.4 g of Rigiruth D-glutamic acid ethyl ester was obtained.

(3) Add 29.0 g of dicyclohexylamine at 25 ° C to a 975 ml suspension of acetonitrile of 81.4 g of the above nicotinol compound at 25 ° C, and then heat to 60 ° C for 2 hours. Stirred. After cooling on ice, the crystals were collected by filtration, washed with acetonitril, dried at 60 ° C, N ⁶ -tert-butoxycarbonyl-N ² -nicotinol-L-ligyl-D-glu Ethyl phosphate ■ 85.7 g of dicyclohexylamine salt was obtained. Melting point 15 7 to 15 9 ° C

(4) 1000 g of the above nicotinoyl dicyclohexylamine salt and (2S, 3aS, 7aS) one-year-old hydro 1H-indole-12-potassium ethyl 345 g of methyl chloride In addition to 1-ethyl-31- (3-diethylaminopropane) carbodiimide hydrochloride 291 under ice-cooling, stir at the same temperature for 1 hour, and then add 2 (16 hours at TC). After the precipitate was removed by filtration, the solvent was distilled off, and the residue was dissolved in ethyl acetate. A 10% aqueous solution of citric acid, a saturated aqueous solution of sodium bicarbonate, and a saturated aqueous solution of sodium chloride were added in that order. After washing and drying over anhydrous sodium sulfate, the solvent was reduced / removed and distilled off (2S.3aS.7aS) -1 (N ^fi — tert-butoxycarbo N ² — Nicotinolium) L one lysyl 10 ¹ - Echiru 'single D- guru evening mil) O click Even 1-H-indole-2-2-ethyl carboxylate 967 g was obtained,

(5) Add 2.0 ^ of tetrahydrofuran to 967 g of the above product, dissolve by heating to 60 ° C, cool on ice, and cool 5N sodium hydroxide at 30 ° C or less. An aqueous solution (850 ml) was added, followed by stirring at 25 ° C for 6 hours. The reaction solution was adjusted to pH 7 with a saturated aqueous solution of citric acid, and tetrahydrofuran was distilled off under reduced pressure at 60 ° C or lower. To the residue was added black form 15 and water 2.5, and the mixture was adjusted to pH 3 with a saturated aqueous solution of citric acid and then shaken. The organic layer was washed with 5% saturated aqueous solution of citric acid and then with saturated aqueous solution of sodium chloride, and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure (2S, 3aS, 7aS) - 1 - (N 6 one ter t-Bed Tokishikarubo two Lou N ² - Nikochino Iru L over Li Giroux § one D- Guru evening mil) O click evening arsenide Draw 1 H—indul 2—Carbonic acid 941 g was obtained.

(6) 4 N hydrochloric acid ethyl acetate 4.0 was added to a suspension of 900 g of the above product in 1.0 ethyl acetate, and the mixture was stirred at 25 ° C for 6 hours. The solvent was distilled off under reduced pressure, and the residue was dissolved in water (3.0), adjusted to pH 7 with concentrated hydrochloric acid, and concentrated again under reduced pressure. The residual solution was adjusted to pH 3 with concentrated hydrochloric acid, applied to a column chromatography using SP-825, and desalted by flowing deionized water. Next, after eluting with 10% acetonitrile / water to remove impurities, the desired fraction eluted with 20% acetonitrile Z water is collected, concentrated under reduced pressure, and crude DU- 1 7 7 7 was obtained. Purity as measured by HPLC was 89.5%.

Example 1

Purification of the crude D U-177 7 by crystallization:

To the crude DU-177 7 obtained in Reference Example 1, 100 ml of evening ethanol was added and dissolved by heating at 60 ° C. This solution was allowed to cool slowly with stirring, and then further cooled with ice. The precipitated crystals were collected by filtration and washed with 10 ml of ethanol to obtain 7.5 g of DU-1777 crude crystals. The purity determined by HPLC was 98.6%. Example 2

Purification of the crude D U—177 7 7 by crystallization:

To 578 g of the crude DU-17777 obtained in Reference Example 2, methanol 5.0 was added, and the mixture was stirred and dissolved at 60 ° C. After the solution was slowly cooled while stirring, the precipitated crystals were collected by filtration and washed with methanol l.Oi? To obtain 533 g of DU-1777 crude crystals. The purity measured with 1% (98.2%).

Example 3

Purification of recrystallized crude crystals of D U—177 7:

(1) To the crude crystals of DU-1777 obtained in Example 1, 7.5 ml of water was added, and the mixture was heated to 50 ° C to dissolve it. Then, 75 ml of methanol was added, and the mixture was added again at 50. Warmed. The mixture was allowed to cool slowly with stirring, and the precipitated crystals were collected by filtration. After drying at 40 ° C for 24 hours, 6.8 g of DU—1 7 7 7 ■ 1 / 2CH ₃ 0H ■ 3 / 2H ₂₀ I got Melting point 199.5-200.5 ° C

As the elemental analysis _{_{_{_{C 26 H 37 N 5 0 7}}}} · 1 / 2CH 3 0H · 3 / 2H 2 0,

Calculated: C, 55.39; H, 7.37; N, 12.19.

Found: C, 55.53; H, 7.25; N, 12.31.

Purification was measured by HPLC DU- 1 7 7 7 purity water 6.8ml added to purified DU- 1 7 7 7 obtained in _c (2 destination was 99.7%, and dissolved by heating to 50 ° C After that, 95 ml of ethanol was added, and the mixture was again dissolved by heating to 60 ° C. The mixture was allowed to cool slowly with stirring, and the precipitated crystals were collected by filtration and then blown at 40 ° C. for 24 hours. dried to give the _{DU- 1 7 7 7 · C 2} H 5 0H '1 / 2H 2 0 of 6.3 g. mp 1 9 8~ 2 0 0 ° C

As the elemental analysis C ₂₆ H ₃₇ N ₅ 〇 Ma _{_{· C 2 H 5 0H · 1}} 2H 2 0,

Calculated: C, 57.32; H, 7.56; N, 11.95.

Found: C, 57.45; H, 7.53; N, 12.01.

Purity of purified DU-17777 measured by HPLC was 99.8%. ₍ Results of powder X-ray diffraction show that the purified DU-17777 obtained here is crystalline. We support being shape.

Example 4

Purification of recrystallized crude crystals of D U-177 7:

550 ml of water was added to the crude crystals of DU-1777 obtained in Example 2, and the mixture was heated to 50 ° C to dissolve the mixture. Then, 5.0 £ of methanol was added and the mixture was heated to 50 ° C again. With stirring was allowed to cool gradually, after the precipitated crystals were滹取, 24 hours air drying to 494 g at 40 ° C DU - 1 7 7 7 · 1 / 2CH 3 0H - give 1 / 2H ₂ 0 Was. Melting point 200 ~ 210 ° C

Purity of purified U U-177 measured by HPLC was 99.9% (industrial availability

The purification method of the present invention is an industrially extremely advantageous method because it provides high-purity DU-1777 applicable as a pharmaceutical product by a simple operation.

Claims

The scope of the claims

1. Crude (2S, 3aS, 7aS) — 1 (N ² —Nicotinyl L-Ligi-Lu ₇ —D—Glu-Mille) Okta-Hydro-1 H-Indole-1 2 —Carboxylic acid and wherein the crystallization child with main Tano Lumpur (2S, 3aS.7aS) - 1 - (N 2 - Nikochino Iru L- lysyl - § one D- guru evening mil) O click evening arsenide A method for purifying 1D-indole1-2-carboxylic acid.

2. (2S.3aS, 7aS) — 1 — (N ^2- Nicotinol 1 L-Lysileu γ-D-Guru Yu Mil) Oku Yu Hydro 1 Η-Indul 2-Carbo It is characterized by recrystallizing crude crystals of acid from a mixed solvent of one or more solvents selected from water, ethylene glycol and propylene glycol and lower alcohols (2S, 3aS, 7aS). — 1 — (N ² nicotinol L L-ジルールーグル 7 — D グル夕ルー）クタクタ 1 1 1 2 2 方法方法方法方法)

3. The purification method according to claim 2, wherein the recrystallization solvent is a mixed solvent of water and methanol.