CRYSTALS, PREPARATION METHOD AND APPLICATION OF A MUSCARINIC RECEPTOR ANTAGONIST
20230167107 · 2023-06-01
Assignee
Inventors
Cpc classification
A61P1/00
HUMAN NECESSITIES
C07D453/02
CHEMISTRY; METALLURGY
International classification
Abstract
The present invention provides crystals of a quaternary ammonium salt structure compound, i.e., (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl) azabicyclo[2,2,2]octylonium bromide (Compound I). A Type-A crystal of Compound I displays diffraction peaks at the following diffraction angles 2θ in a X-ray powder diffraction pattern thereof: 5.7±0.2 degrees, 12.9±0.2 degrees, 16.7±0.2 degrees, 18.0±0.2 degrees, 19.5±0.2 degrees, 21.1±0.2 degrees, 22.3±0.2 degrees and 23.3±0.2 degrees. A Type-B crystal of Compound I displays diffraction peaks at the following diffraction angles 2θ in a X-ray powder diffraction pattern thereof: 5.2±0.2 degrees, 15.8±0.2 degrees, 16.9±0.2 degrees, 17.7±0.2 degrees, 19.5±0.2 degrees, 20.2±0.2 degrees and 22.1±0.2 degrees. The present application also relates to a new method for preparing Compound I and applications of the two novel crystals in the field of medicine.
Claims
1. A Type-A crystal of (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide, displaying diffraction peaks at the following diffraction angles 2θ in an X-ray powder diffraction pattern: 5.7±0.2 degrees, 12.9±0.2 degrees, 16.7±0.2 degrees, 18.0±0.2 degrees, 19.5±0.2 degrees, 21.1±0.2 degrees, 22.3±0.2 degrees, and 23.3±0.2 degrees, wherein the X-ray powder diffraction pattern is a spectrum obtained by using Cu Kα rays.
2. A Type-B crystal of (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide, displaying diffraction peaks at the following diffraction angles 2θ in an X-ray powder diffraction pattern: 5.2±0.2 degrees, 15.8±0.2 degrees, 16.9±0.2 degrees, 17.7±0.2 degrees, 19.5±0.2 degrees, 20.2±0.2 degrees, and 22.1±0.2 degrees, wherein the X-ray powder diffraction pattern is a spectrum obtained by using Cu Kα rays.
3. A method for preparing (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide, comprising steps of: (1) reducing a starting material cyclopentyl mandelic acid or cyclopentyl mandelate by sodium borohydride to obtain racemic 2-hydroxy-2-cyclopentyl-2-phenylethanol (Z02), with solvent selected from a group consisting of dimethoxyethane, tetrahydrofuran, dioxane, methanol and ethanol, wherein the sodium borohydride and the starting material have a molar ratio of 2:1 to 5:1; a Lewis acid is added for catalysis when reducing the cyclopentyl mandelic acid, wherein the Lewis acid is selected from a group consisting of aluminum trichloride, boron trifluoride, zinc chloride, tin tetrachloride and titanium tetrachloride, and the Lewis acid and the cyclopentyl mandelic acid have a molar ratio of 2:1 to 5:1, preferably 2.5:1 to 3:1; (2) reacting Z02 with chiral acyl chloride to perform an esterification reaction and obtain chiral 2-hydroxy-2-cyclopentyl-2-phenylethanol carboxylate (Z03) as a crystal, wherein the chiral acyl chloride is selected from a group consisting of L-camphorsulfonyl chloride, D-camphorsulfonyl chloride and acyl chloride derivatives of mandelic acid; Z02 and the chiral acyl chloride have a molar ratio of 1:1 to 1:3, preferably 1:1.5 to 1:2; solvent is selected from a group consisting of dichloromethane, chloroform, tetrahydrofuran and dioxane, preferably dichloromethane and tetrahydrofuran; base is selected from a group consisting of triethylamine, pyridine and N-methylmorpholine, and the base and the chiral acyl chloride have a molar ratio of 1:1 to 4:1, preferably 1:1 to 2:1; (3) treating Z03 with a base to obtain R-1-phenyl-1-cyclopentyl oxirane (Z04) wherein the base is selected from a group consisting of sodium hydride, potassium tert-butoxide, butyl lithium and sodium amide, preferably sodium hydride and potassium tert-butoxide; the base and Z03 have a molar ratio of 1:1 to 3:1, preferably 1:1 to 1.5:1; solvent is selected from a group consisting of dichloromethane, tetrahydrofuran, dioxane and dimethyl sulfoxide, preferably dimethyl sulfoxide and tetrahydrofuran; (4) reacting Z04 with (R)-(−)-3-quinuclidinol to obtain free base of (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-azabicyclo[2,2,2]octane (Z05), wherein the base includes but is not limited to sodium hydride, potassium tert-butoxide, butyl lithium and sodium amide, preferably sodium hydride and potassium tert-butoxide; (R)-(−)-3-quinuclidinol and the base have a molar ratio of 1:1 to 3:1, preferably 1:1 to 1.5:1; solvent is selected from a group consisting of dichloromethane, tetrahydrofuran, dioxane and dimethyl sulfoxide, preferably dimethyl sulfoxide and tetrahydrofuran; and (5) reacting Z05 with 3-phenoxy-1-bromopropane (Z06) to perform a quaternization reaction and to obtain (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl) ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide.
4. A method for preparing the Type-A crystal of (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide according to claim 1, comprising the steps of: heating to dissolve (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide in a good solvent selected from a group consisting of alcohols, acetonitrile, dichloride methane and chloroform; adding an anti-solvent to the resulting solution, wherein the anti-solvent is selected from a group consisting of esters, ethers, ketones, liquid cycloalkanes and aromatic hydrocarbons; and slowly cooling the resulting solution for crystallization to obtain the Type-A crystal with avoiding exposure to moisture during the entire process.
5. A method for preparing the Type-B crystal of (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide according to claim 2, comprising the steps of: heating to dissolve (2R,3R)-3-[(2-cyclopentyl-2-hydroxy phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide in an alcohol or a mixed solvent of acetonitrile and water, preferably ethanol; adding an anti-solvent to the resulting solution, wherein the anti-solvent is selected from a group consisting of esters, water, ethers, ketones, liquid cycloalkanes and aromatic hydrocarbons, preferably ethyl acetate; slowly cooling the resulting solution for crystallization to obtain the Type-B crystal; or heating to dissolve (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide in a solvent selected from a group consisting of alcohols, acetonitrile, dichloride methane and chloroform; adding water to the resulting solution; and slowly cooling the resulting solution for crystallization to obtain the Type-B crystal.
6. A method for preparing the Type-B crystal of (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide according to claim 2, comprising the steps of: adding a Type-A crystal of (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide to a reaction vessel; adding purified water and stirring to form a slurry; filtering by suction filtration; and air-drying the resulting solid at 40° C. to 80° C. to reach a constant weight, thereby obtaining the Type-B crystal.
7. A pharmaceutical composition comprising the Type-A crystal of claim 1 as an active ingredient.
8. The use of the Type-A crystal of claim 1 for preparing an M receptor subtype selective antagonist.
9. The use of the Type-A crystal of claim 1 for preparing preventive or therapeutic agents in applications of rhinitis, post-cold rhinitis, chronic bronchitis, airway hyperactivity, asthma, COPD, cough, urinary incontinence, frequent urination, unstable bladder syndrome, bladder spasm, cystitis, and gastrointestinal diseases such as irritable bowel syndrome, spastic colitis, and duodenal and gastric ulcers.
10. A pharmaceutical composition comprising the Type-B crystal of claim 2 as an active ingredient.
11. The use of the Type-B crystal of claim 2 for preparing an M receptor subtype selective antagonist.
12. The use of the Type-B crystal of claim 2 for preparing preventive or therapeutic agents in applications of rhinitis, post-cold rhinitis, chronic bronchitis, airway hyperactivity, asthma, COPD, cough, urinary incontinence, frequent urination, unstable bladder syndrome, bladder spasm, cystitis, and gastrointestinal diseases such as irritable bowel syndrome, spastic colitis, and duodenal and gastric ulcers.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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DETAILED DESCRIPTIONS
[0103] The present invention will be illustrated in more detail with reference to the following examples and test examples, but those skilled in the art know that the present invention is not limited to these examples and test examples.
Example 1
Preparation of Compound I (Raw Pharmaceutical Material) of the Present Invention
Step 1: Preparation of 2-hydroxy-2-cyclopentyl-2-phenylethanol
[0104] Method 1:
[0105] 124.0 g (0.500 mol) of ethyl cyclopentyl mandelate and 1000 mL of anhydrous ethanol were added into a 2 L three-necked flask. The flask was then put into an ice-salt bath until internal temperature below 5° C. 37.83 g (1.00 mol) of sodium borohydride was added in batches while the internal temperature was kept not to exceed 5° C. Then, the internal temperature was raised to about 45° C. and react for 2 h. The solvent was removed under reduced pressure after reaction was completed. The obtained residue was neutralized to neutrality with 0.5 M hydrochloric acid and extracted three times with dichloromethane (3×500 mL). Organic phases were combined and dried with anhydrous magnesium sulfate. The desiccant was then filtered off and the solvent was completely removed from the filtrate under reduced pressure, to obtain 98.6 g of yellow oil (yield: 95.60%) which was used directly in the next reaction.
[0106] Method 2:
[0107] 200.00 g (0.908 mol) of cyclopentyl mandelic acid and 3000 mL of ethylene glycol dimethyl ether were added into a 5 L three-necked flask. The flask was then put into an ice-salt bath until internal temperature below 0° C. 363.20 g (2.724 mol) of aluminum trichloride was added while the internal temperature was kept not to exceed 5° C. After completing the addition, the mixture was stirred and reacted at its temperature for half an hour, and then 137.40 g (3.632 mol) of sodium borohydride was added in batches while the internal temperature was kept not to exceed 5° C. Then, the internal temperature was raised to about 55° C. to react for 2 h. Thin-layer chromatography (TLC) detection was performed to confirm completion of the reaction. The reaction mixture was slowly poured into 1600 mL of ice-cold 1 mol/L hydrochloric acid under constant stirring. Temperature of the solution was controlled not to exceed 25° C. Then the solution was extracted three times with ethyl acetate (3×1000 mL) and organic phases were combined and washed three times with 5% sodium carbonate aqueous solution (3×500 mL). The organic phase was separated and washed three times with 5% sodium chloride aqueous solution (3×500 mL). The organic phase was dried with anhydrous magnesium sulfate. The desiccant was filtered off and the solvent was removed at 50° C. under reduced pressure. The residue was dissolved with isopropyl ether (1000 mL), and then was washed with 2 mol/L sodium hydroxide aqueous solution (450 mL) under mechanical stirring for 10 min. The mixture was then placed in a separatory funnel and the organic layer was separated and dried by adding anhydrous magnesium sulfate. The desiccant was filtered off, and the solvent was removed by rotary evaporation under reduced pressure with a water pump, to obtain 145.80 g of light yellow oil (yield: 77.84%) which was used directly in the next reaction.
Step 2: Preparation of (R)-2-hydroxy-2-cyclopentyl-2-phenethanolylL-(−)-camphorsulfonyl
[0108] 112.50 g (545.38 mmol) of 2-hydroxy-2-cyclopentyl-2-phenylethanol was dissolved in 700 mL of dichloromethane and the obtained solution was poured into a 5 L three-neck reaction flask. After the solution became clear and transparent, 165.55 g (1636.03 mmol) of triethylamine was added at room temperature and then the flask was then put into an ice-water bath until an internal temperature thereof below 10° C. 500 mL of dichloromethane solution containing 164.10 g (654.46 mmol) of L-(−)-camphorsulfonyl chloride was added dropwise. After completing the addition, the internal temperature was raised to about 10° C. to react for 1 h. When the reaction is completed, 1 L of water was added and the mixture was placed in a separatory funnel. Aqueous phase and organic phase were separated. The organic phase was washed three times with water for 1 L each time (3×1000 mL). Finally, the organic phase was collected and dried by adding anhydrous magnesium sulfate. The desiccant was filtered off, and the solvent was removed by rotary evaporation at 40° C. under reduced pressure. The residue was dissolved in 200 mL of ethyl acetate, and crystallized by freezing. The solid was collected by filtration and dried to obtain 70.03 g of a white solid (yield: 61.0%).
Step 3: Preparation of (R)-2-cyclopentyl-2-phenyl oxirane
[0109] Dimethyl sulfoxide (350 mL) and 2-hydroxy-2-cyclopentyl-2-phenethanolyl L-(−)-camphorsulfonyl (69.23 g, 164.6 mmol) were added into a 1 L three-necked reaction flask. After the solution was clear, potassium tert-butoxide (17.54 g, 156.31 mmol) was added at room temperature, and the flask was then put into an oil bath to heat to an internal temperature of 50° C. After reacting for 1 h, TLC detection was performed (developing solvent: petroleum ether:ethyl acetate=3 mL:1 mL). The mixture was put into an ice-water bath until an internal temperature thereof below 10° C., and 450 mL of water was added dropwise. The resulting mixture was placed in a separatory funnel, and extracted three times with isopropyl ether (3×200 mL). The organic phases obtained were combined, washed three times with 5% sodium chloride solution (3×200 mL), and dried by adding anhydrous magnesium sulfate. The desiccant was filtered off, and the filtrate was rotary evaporated under reduced pressure in a water bath at 45° C. to remove the solvent. 28.02 g of a light yellow liquid was finally obtained (yield: 90.42%).
Step 4: Preparation of (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-azabicyclo[2,2,2]octane
[0110] R-(−)-3-quinuclidinol (16.46 g, 129.45 mmol) and 250 mL of anhydrous tetrahydrofuran were added into a 1 L three-neck reaction flask. When the solid was dissolved, Z04 (24.38 g, 129.45 mmol dissolved in 50 mL of tetrahydrofuran) was added at room temperature. The flask was put into an oil bath to heat to an internal temperature of 85° C. Then, NaH (60%, 3.45 g, 86.3 mmol) was added, and the reaction was kept at 85° C. for 2 h. TLC detection (developing solvent: petroleum ether:ethyl acetate=5:0.1) was performed. When the reaction was completed, the solvent was removed under reduced pressure, and 500 mL of ice water was added dropwise to the residue, followed by extraction 3 times with ethyl acetate (3×500 mL). The organic phases obtained were combined and dried with anhydrous magnesium sulfate. The desiccant was removed by filtration, and the solvent was removed under reduced pressure to obtain a light yellow-brown solid. The solid was dissolved with isopropyl ether by heating and refluxing, and crystallized by freezing. The resulting solid was collected by filtration and dried to constant weight. Finally, 36.28 g of an off-white solid as the target compound was obtained (yield: 88.85%).
Step 5: (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octylonium bromide
[0111] 98.46 g (0.312 mol) of (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-azabicyclo[2,2,2]octane and 490 mL of anhydrous ethanol were placed in a 5 L reactor, and the mixture obtained was stirred until dissolution at room temperature. When the dissolution is complete, about 100 mL of anhydrous ethanol solution containing 82.93 g (0.386 mol) of 3-phenoxy bromopropane was added, and the mixture was heated to reflux for 2 h. When the reaction was completed, the solvent was removed under reduced pressure to obtain 149.28 g of Compound I as an off-white solid (yield: 90.2%).
[Example 2] Preparation of Type-A Crystal of the Present Invention
[0112] 43.3 g of Compound I (prepared according to Example 1, also referred to as raw pharmaceutical material, similarly hereinafter) and 86.7 mL of anhydrous ethanol were weighed and put into a 200 mL eggplant-shaped flask. The mixture was refluxed and stirred to complete dissolution. Then, 0.87 g of activated carbon was added. The mixture was refluxed and stirred for 0.5 h for decoloration. The activated carbon was removed by suction filtration while the mixture was still hot, to obtain a light yellow transparent filtrate. The filtrate was transferred into a 1.0 L eggplant-shaped flask, and 1.0 L of ethyl acetate was added under reflux conditions. The mixture was cooled to 25±5° C. and stirred for 2 h to crystallize. The resulting solid was filtered by suction filtration and dried by blowing air at 80° C. for 4 h, to obtain Type-A crystals of the present invention (39.2 g, yield: 90.5%). The X-ray powder diffraction pattern of Type-A crystal of the present invention is shown in
[0113] Elemental analysis C.sub.29H.sub.40BrNO.sub.3, calculated values C, 65.65, H, 7.60, Br, 15.06, N, 2.64; measured values C, 65.60, H, 7.58, Br, 15.0, N, 2.62. Elemental analysis results show that the product does not contain crystal water or other crystal solvents, which is consistent with the molecular formula of Compound I.
[Example 3] Preparation of Type-A Crystal of the Present Invention
[0114] 35.1 g of Compound I and 300 mL of dichloromethane were weighed and put into a 500 mL eggplant-shaped flask. The mixture was refluxed and stirred until dissolution. Then, 0.7 g of activated carbon was added. The mixture was refluxed and stirred for 2 h for decoloration. The activated carbon was removed by suction filtration while the mixture was still hot, to obtain a light yellow transparent filtrate. The filtrate was transferred into a 2.0 L eggplant-shaped flask. Under reflux conditions, 3 g of Crystal A was added, and then 900 L of isopropyl ether was added. The mixture was cooled to 5±5° C. and stirred for 48 h to crystallize. The resulting solid was filtered by suction filtration and dried under conditions of 40° C. and 10 mmHg or less for 24 h, to obtain Type-A crystal of the present invention (27.6 g, yield: 72.3%).
[0115] Elemental analysis C.sub.29H.sub.40BrNO.sub.3, calculated values C, 65.65, H, 7.60, Br, 15.06, N, 2.64; measured values C, 65.45, H, 7.42, Br, 15.11, N, 2.50. Elemental analysis results show that the product does not contain crystal water or other crystal solvents, which is consistent with the molecular formula of Compound I.
[Example 4] Preparation of Type-B Crystal of the Present Invention
[0116] 30.3 g of Compound I (prepared according to Example 1, also referred to as raw pharmaceutical material, similarly hereinafter) and 100 mL of 95% ethanol were weighed and put into a 200 mL eggplant-shaped flask. The mixture was refluxed and stirred until dissolution. Then, 0.6 g of activated carbon was added. The mixture was refluxed and stirred for 2 h for decoloration. The activated carbon was removed by suction filtration while the mixture was still hot, to obtain a light yellow transparent filtrate. The filtrate was transferred into a 2.0 L eggplant-shaped flask, and 1000 mL of tetrahydrofuran was added under reflux conditions. The mixture was cooled to 20±5° C. and stirred for 2 h to crystallize. The resulting solid was filtered by suction filtration and dried to constant weight by blowing air at 60° C. for 8 h, to obtain Type-B crystal of the present invention (26.5 g, yield: 83.3%). The X-ray powder diffraction pattern of Type-B crystal of the present invention is shown in
[0117] Elemental analysis proved that Type-B crystal of Compound I contains 1.5 crystal water, with a molecular formula of C.sub.29H.sub.40BrNO.sub.3.1.5H.sub.2O, calculated values C, 62.47, H, 7.77, Br, 14.33, N, 2.51; measured values C, 62.60, H, 7.82, Br, 14.25, N, 2.48.
[Example 5] Preparation of Type-B Crystal of the Present Invention
[0118] 32.8 g of Compound I and 200 mL of 98% ethanol were weighed and put into a 500 mL eggplant-shaped flask. The mixture was refluxed and stirred until dissolution. Then, 0.7 g of activated carbon was added. The mixture was refluxed and stirred for 4 h for decoloration. The activated carbon was removed by suction filtration while the mixture was hot, to obtain a light yellow transparent filtrate. The filtrate was transferred into a 5.0 L reactor, and 1000 mL of acetone was added under reflux conditions. The mixture was cooled to 10±5° C. and stirred for 24 h to crystallize. The resulting solid was filtered by suction filtration and dried to constant weight by blowing air at 80° C. for 4 h, to obtain Type-B crystal of the present invention (25.5 g, yield: 74.9%).
[0119] Elemental analysis proved that Type-B crystal of Compound I contains 1.5 crystal water, with a molecular formula of C.sub.29H.sub.40BrNO.sub.3.1.5H.sub.2O, calculated values C, 62.47, H, 7.77, Br, 14.33, N, 2.51; measured values C, 62.36, H, 7.79, Br, 14.21, N, 2.68.
[Example 6] Preparation of Type-B Crystal by Converting Crystal Method
[0120] 40.3 g of crystal A of Compound I was weighed and added into a 1 L reactor. 400 mL of purified water was added, and stirred for 5 h with a rotate speed of 250-270 r/min at 25±5° C. The resulting solid was filtered by suction filtration and dried by blowing air at 60° C. for 12 h. When the weight of water was detected as constant, Type-B crystal of the present invention (35.7 g, yield 84.3%) was obtained. The X-ray powder diffraction pattern of Type-B crystal of the present invention is shown in
[0121] Elemental analysis proved that Type-B crystal of Compound I contains 1.5 crystal water, with a molecular formula of C.sub.29H.sub.40BrNO.sub.3.1.5H.sub.2O, calculated values C, 62.47, H, 7.77, Br, 14.33, N, 2.51; measured values C, 62.41, H, 7.84, Br, 14.16, N, 2.63.
[Preparation Example 7] Preparation of Dry Powder Inhalation Composition for Maintenance Treatment of Asthma and COPD
[0122] Components in the Composition and Amounts
[0123] Crystal A 100 mg
[0124] Lactose 25000 mg
[0125] Crystal A of the present invention was micronized to an average particle size D50 of less than 5 μm. The micronized Crystal A was thoroughly mixed with lactose having a particle size of 1 μm to 100 μm, and the mixture was filled into capsules. The amount of the drug/lactose mixture per capsule is 25.1 mg, which is administered by powder inhaler.
TEST EXAMPLES
[0126] Type-A crystal of the present invention was prepared by using methods of Example 2 or based the same mechanism as Example 2; Type-B crystal of the present invention was prepared by using methods of Example 4 or 6, or based the same mechanism as Example 4 or 6.
[Test Example 1] X-Ray Powder Diffraction (XRPD) Test of Crystals of the Present Invention
[0127] The solid samples obtained were analyzed with an X-ray powder diffraction analyzer (Bruker D8 advance) equipped with a LynxEye detector. The samples was scanned with a 20 scanning angle from 3° to 40°, a scanning step of 0.02°, a tube voltage of 40 KV and a tube current of 40 mA. The sample pan used for sample measurement is a zero background sample pan.
[0128] Results:
[0129] (1) The result of Type-A crystal is shown in
TABLE-US-00001 TABLE 1 X-ray powder diffraction data of Crystal A of Example 2 Diffraction D value Relative angle (2θ) (Å) strength (%) 5.698 15.497 100 8.571 10.309 6.9 12.875 6.870 12.9 16.068 5.512 3.7 16.785 5.278 19 17.963 4.934 23.9 19.544 4.538 12.3 20.137 4.406 4.7 21.107 4.206 20.9 22.273 3.988 19.1 23.272 3.819 12.9 23.844 3.729 3.8 24.275 3.664 6.6 25.132 3.541 2.7 25.595 3.478 6.3 25.787 3.452 4.2 27.666 3.222 4.2 28.039 3.180 9.3 28.76 3.102 8.3 29.678 3.008 8.4 30.827 2.898 7 34.323 2.611 8.9 34.946 2.565 3 35.128 2.553 3 39.227 2.295 3.2 39.549 2.277 2.9
[0130] About 10 mg of Crystal A was weighed in an 8 mL sample bottle. The bottle was sealed with filter paper, and put into a 40° C./75% RH chamber for stability test. After 3 days, the sample was taken out for XRPD detection. The result showed that Crystal A had been partially converted into Crystal B, indicating that Type-A crystal was unstable under high humidity conditions.
[0131] (2) The result of Type-B crystal is shown in
TABLE-US-00002 TABLE 2 Characteristic X-ray powder diffraction data of Crystal B of Example 6 Diffraction D value Relative angle (2θ) (Å) strength (%) 5.216 16.928 100 7.921 11.153 2.2 11.05 8.0 2.6 13.015 6.797 4.8 15.13 5.851 2.8 15.758 5.619 25.9 16.871 5.251 9.6 17.35 5.107 3.5 17.681 5.012 7.1 18.652 4.754 4.4 19.507 4.547 9.8 19.815 4.477 6.1 20.219 4.388 8.8 21.927 4.050 8.6 22.123 4.015 13.2 22.946 3.872 3 23.276 3.819 2.8 24.584 3.618 2.6 25.193 3.532 2.6 25.735 3.459 2.8 26.932 3.308 5.5 28.253 3.156 3.8 32.839 2.725 2.9 37.334 2.407 3.1
[0132] About 10 mg of Crystal B was weighed in an 8 mL sample bottle. The bottle was sealed with filter paper, and put into a 40° C./75% RH chamber for stability test. After 3 days, the sample was taken out for XRPD detection. The result showed that Crystal B did not change.
[0133] Test Example 1 shows that Type-B crystal is more stable than Type-A crystal in a high-humidity environment.
[Test Example 2] Thermo-Gravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC) Test
[0134] TGA: The solid samples were analyzed with TA TGA Q500 for thermo-gravimetric analysis. 2 mg to 3 mg sample was placed in a balanced aluminum sample pan, and the mass of the sample was automatically weighed in the TGA heating furnace. The sample was heated to 200° C. to 300° C. at a temperature increase rate of 10° C./min. During the test, flow rates of nitrogen to the balance chamber and the sample chamber were 40 mL/min and 60 mL/min, respectively.
[0135] DSC: The solid samples were analyzed with TA DSC Q200 for differential scanning calorimetry analysis, with indium used as the standard sample for calibration. 2 mg to 3 mg sample was accurately weighed and placed in the TA DSC sample pan, and the accurate mass of the sample was recorded. The sample was heated to 200° C. to 250° C. at a temperature increase rate of 10° C./min in a nitrogen flow of 50 mL/min.
[0136] Results: TGA results are shown in
[0137] DSC results are shown in
[Test Example 3] Dynamic Vapor Sorption (DVS)
[0138] The dynamic vapor sorption and desorption analysis was performed on the IGA SORP (Hidenlsochema) instrument. The samples were tested in gradient mode. The test was performed in a humidity range of 0% to 90%, with an incremental humidity between each gradient of 10%. For each gradient, the shortest test time was 30 min, and the longest test time was 120 min. There is a time interval of 3 min for collecting data in the system.
[0139] Result: Results of DVS are shown in
[Test Example 4] Content Measurement of the Residual Solvent Contained in the Crystals of the Present Invention
[0140] The content of the residual solvent contained in the crystal of the present invention was measured using the following test conditions. The results are shown in Table 3.
[0141] Test Conditions:
[0142] Gas chromatography equipped with FID detector
[0143] Chromatographic column: OPTIMA-624 (30 m×0.32 mm×1.8 μm)
[0144] Column temperature: 60° C. (3 min) 20° C./min 200° C. (5 min)
[0145] Inlet temperature: 200° C.
[0146] Detector temperature: 250° C.
[0147] Carrier gas: Nitrogen
[0148] Column flow rate: 2 mL/min
[0149] Headspace parameters: headspace balance temperature 80° C. [0150] headspace balance time 30 min
[0151] Split ratio: 10:1
TABLE-US-00003 TABLE 3 Residual solvents in the pharmaceutical raw material and crystals of the present invention Content Crystalline form Solvent (ppm) Example 1, Raw Ethanol 21371 pharmaceutical Isopropyl ether 3826 material Example 2, Crystal Type-A crystal of the Ethanol 209 present invention Example 5, Crystal Type-B crystal of the Ethanol 85 present invention Example 6, Crystal Type-B crystal of the Not present invention detected
[0152] The residual solvent was removed in each recrystallization process of the present invention. Both Type-A crystal and Type-B crystal of the present invention have very little residual solvent. In Example 6, no residual solvent was detected in the crystal.
[Test Example 5] Impurity Removal Effect of Recrystallization
[0153] The following conditions were used in high-performance liquid chromatography to determine the impurity removal effect of the recrystallization process of the crystals of the present invention.
[0154] Instrument: High performance liquid chromatograph equipped with UV detector
[0155] Chromatographic column: AgelaPromosil C18 4.6×250 mm, 5 μm
[0156] Mobile phase: Phase A: 0.01 mol/L potassium dihydrogen phosphate solution (added with 0.04 mol/L of ammonium chloride, and the pH was adjusted to 3.0 by using phosphoric acid)-methanol (38:62); Phase B: acetonitrile
[0157] Gradient elution table:
TABLE-US-00004 Time (min) Phase A (%) Phase B (%) 0 100 0 30 75 25 60 40 60 65 40 60 68 100 0 80 100 0
[0158] Detection wavelength: 210 nm
[0159] Flow rate: 1.0 mL/min
[0160] Injection volume: 20 μL
[0161] Column temperature: 30° C.
[0162] Solvent: mobile phase A
[0163] First, the purity (%) of Compound I in each crystal was calculated according to HPLC chromatography by the following formula: Purity (%) of Compound I in each crystal=(Peak area of Compound I in each crystal)/(Total area of all peaks)×100.
[0164] Next, the impurity removal rate (%) in each crystal was calculated by the following formula: Impurity removal rate in each crystal (%)=[{(Purity of Compound I in each crystal)−(Purity of Compound I in pharmaceutical raw material)}/{100−(Purity of Compound I in pharmaceutical raw material)}]×100
[0165] The results are shown in Table 4.
TABLE-US-00005 TABLE 4 Results of recrystallization to remove impurities in pharmaceutical raw materials Purity of Com- Impurity removal pound I in each rate in each No. Crystalline form crystal form (%) crystal form (%) 1 Raw pharmaceutical 98.15 — material 2 Type-A crystal of the 99.78 88.1 present invention 3 Type-B crystal of the 99.82 90.3 present invention
[0166] The results show that the recrystallization processes for preparing Type-A and Type-B crystals of the present invention can remove most of the impurities in the raw pharmaceutical materials.
[Test Example 6] Research on the Crystallization Solvent
[0167] According to “2. Preparation of Type-A crystal and Type-B crystal of the present invention (hereinafter collectively referred to as the crystal of the present invention)” of “Best mode”, the recrystallization method for the Type-A and Type-B crystals of the present invention sometimes could not precipitate any crystal under certain conditions, even if the solvent is removed, the raw pharmaceutical material may eventually become an oil. In the preparation of Type-A crystal, good solvents are alcohol and acetonitrile at 2° C. to 4° C., and anti-solvents are tetrahydrofuran and methyl tert-butyl ether for this phenomenon. In the preparation of Type-B crystal, good solvents are alcohol/water solution and acetonitrile at 2° C. to 4° C., and anti-solvents are tetrahydrofuran and methyl tert-butyl ether for this phenomenon. 12 kinds of good solvents (including good solvents containing a certain amount of water) and 8 kinds of anti-solvents were used to carry out a total of 192 crystallization combinations. It was found that no crystal was obtained in 9 cases in the following table, and the raw pharmaceutical material became oily substance (oil) after the solvent was evaporated.
TABLE-US-00006 TABLE 5 Cases in which no crystal is obtained by recrystallization Expected crystal Actual No. Good solvent (ratio 1*) Anti-solvent (ratio 2.sup.‡) form result 1 Ethanol (1) Tetrahydrofuran (1) Type-A crystal Oil 2 Ethanol (2) Methyl tert-butyl ether (2) Type-A crystal Oil 3 Isopropanol (2) Tetrahydrofuran (3) Type-A crystal Oil 4 Isobutanol (15) Tetrahydrofuran (1) Type-A crystal Oil 5 Acetonitrile (2) Tetrahydrofuran (2) Type-A crystal Oil 6 Water/Ethanol 95% (2) Methyl tert-butyl ether (5) Type-B crystal Oil 7 Water/Isopropanol 95% Tetrahydrofuran (2) Type-B crystal Oil (2) 8 Water/Isobutanol 97% Tetrahydrofuran (4) Type-B crystal Oil (3) 9 Water/n-Butanol 98% (2) Methyl tert-butyl ether (1) Type-B crystal Oil *Ratio 1: ratio of good solvent/pharmaceutical raw material (mL/g) .sup.‡Ratio 2: ratio of anti-solvent/good solvent (mL/mL)
[0168] It was found that increasing the ratio of anti-solvent/good solvent to a level of >8 can effectively avoid the result of inability to recrystallize.