Salt of cyclohexane derivative
11584731 · 2023-02-21
Assignee
- Shanghai Jingxin Biology & Pharmaceutical Co., Ltd (Shanghai, CN)
- Zhejiang Jingxin Pharmaceutical Co., Ltd. (Shaoxing, CN)
Inventors
- Yue Huang (Zhejiang, CN)
- Fei Zheng (Zhejiang, CN)
- Xiaoyun Fu (Zhejiang, CN)
- Chunlan Tang (Zhejiang, CN)
- Dan Zhu (Zhejiang, CN)
Cpc classification
A61P25/28
HUMAN NECESSITIES
A61P25/18
HUMAN NECESSITIES
A61K31/496
HUMAN NECESSITIES
C07D333/54
CHEMISTRY; METALLURGY
C07D333/66
CHEMISTRY; METALLURGY
International classification
C07D333/66
CHEMISTRY; METALLURGY
A61K31/496
HUMAN NECESSITIES
Abstract
The present invention provides a maleate, phosphate, sulfate, hydrochloride of a cyclohexane derivative, N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, as shown in Formula I and crystal forms thereof. The crystal forms have low hygroscopicity and good stability and are convenient for long-term storage and transportation; or the crystal forms have a long half-life in vivo, high bioavailability and small individual difference, and thus have obvious clinical application advantages.
Claims
1. A salt of a cyclohexane derivative N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea of Formula I, wherein the salt comprises an anion, and the anion is a maleate ion, ##STR00014## the stoichiometric ratio of the compound of Formula I to the anion is 1:1, wherein the X-ray powder diffraction pattern of the salt, expressed in 2θ angle by using Cu-Kα radiation, has diffraction peaks at least at 11.804°±0.2°, 12.703°±0.2°, 13.493°±0.2°, 14.495°±0.2°, 15.096°±0.2°, 17.108°±0.2°, 19.104°±0.2° 19.655°±0.2°, 20.023°±0.2°, 21.6110±0.20 and 24.0880±0.20.
2. The salt of claim 1, wherein the salt also has diffraction peaks at 2θ values of 7.246°±0.2°, 17.567°±0.2°, 18.794°±0.2°, 20.395°±0.2°, 21.030°±0.2°, 22.496°±0.2°, 24.867°±0.2° and 26.412°±0.2°.
3. The salt of claim 2, wherein the salt also has diffraction peaks at 2θ values of 11.045°±0.2°, 22.997°±0.2°, 25.336°±0.2°, 27.786°±0.2°, 28.292°±0.2°, 28.914°±0.2°, 29.804°±0.2°, 30.770°±0.2°, 31.628°±0.2° and 33.952°±0.2°.
4. The salt of claim 1, wherein in DSC analysis, the salt has an endothermic peak at 191.8° C.; in TGA analysis, the weight loss reaches 0.41% at 150° C.
5. A pharmaceutical composition for the treatment or amelioration of schizophrenia or bipolar disorders, wherein the pharmaceutical composition comprises the salt of claim 1 and pharmaceutically acceptable auxiliaries.
6. The salt of claim 3, wherein the salt has an XRPD pattern as shown in
7. The salt of claim 4, wherein the DSC pattern is as shown in
8. The salt of claim 4, wherein the TGA pattern is as shown in
Description
BRIEF DESCRIPTION OF THE DRAWINGS
(1)
(2)
(3)
(4)
(5)
(6)
(7)
(8)
(9)
(10)
(11)
(12)
(13)
(14)
(15)
(16)
(17)
(18)
(19)
(20)
(21)
(22)
(23)
(24)
(25)
(26)
(27)
(28)
(29)
(30)
(31)
(32)
(33)
(34)
(35)
(36)
(37)
(38)
(39)
(40)
(41)
(42)
(43)
(44)
DETAILED DESCRIPTION OF THE EMBODIMENTS
(45) The invention is further illustrated by the following embodiments. It is to be understood that these embodiments are for illustrative purposes only and are not intended to limit the invention. Various changes and modifications made by those skilled in the art based on the concept of the present invention should fall within the protection scope of the present invention.
(46) The X-ray powder diffraction patterns of crystal forms of the various salts of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea of the present invention, are expressed as diffraction peak positions, i.e., diffraction angle 2θ (°), interplanar spacing d (Å), Diffraction peak relative intensity (I/I0).
(47) The term “relative intensity” refers to the ratio of the intensity of the other peak to the intensity of the highest intensity peak when the intensity of the highest intensity peak of all diffraction peaks of the X-ray powder diffraction pattern is 100%.
(48) The term “substantially the same” means that at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% of the peaks in the X-ray powder diffraction pattern appear in the given example X-ray powder diffraction pattern.
EXAMPLES
(49) Reagents: the reactants and the catalyst used in the embodiment of the invention are chemical pure, and can be directly used or simply purified as required; The organic solvents and the like are all analytical reagent and are directly used. All reagents were purchased from Sinopharm Chemical Reagent Co., Ltd (Group) in Shanghai.
(50) The amorphous form of the compound of Formula I is prepared according to the methods reported in the literature, e.g. in Example 5 of CN 106518841A, and is not limited thereto.
(51) X-Ray Powder Diffraction:
(52) X-ray powder diffraction analysis was performed on an X-ray powder diffractometer manufactured by PANalytacal using Cu-Kα radiation, testing a continuous scan of θ-2θ with a power of 45 kV×40 mA, a step width of 0.02°, and a scan range of 3-40° (2θ).
(53) Differential Scanning Calorimetry (DSC) Characterization:
(54) using TA Q2000/2500 differential scanning calorimeter to determine under the condition that the protective gas is nitrogen, the heating rate is 10° C./min, and the temperature rises gradually from 25° C. to the end point.
(55) Thermogravimetric Analysis (TGA):
(56) using TA Q5000/5500 thermal gravimetric analyzer to determine under the condition that the protective gas is nitrogen, the heating rate is 10° C./min, and the temperature rises gradually from room temperature to the end point.
(57) Content Detection Method (HPLC):
(58) Chromatographic Conditions
(59) TABLE-US-00001 Chromatographic Agilent Eclipse Plus C18 column type 4.6 * 100 mm, 3.5 μm Mobile phase A 0.1% FA (formic acid) aqueous solution Mobile phase B MeOH Column temperature (° C.) 40 Flow rate (ml/min) 1.0 Detection wavelength (nm) 260 Sample injection 10 μl amount Specimen disc Room temperature temperature (° C.) Running gradient Time Mobile phase Mobile phase (min) A (%) B (%) 0 90 10 25 10 90 30 10 90 30.1 90 10 40 90 10
Solution Preparation
(60) Diluent (blank solution): acetonitrile/water=1/1 (V/V)
(61) Test sample solution: 5 mg of the sample was precisely weighed and placed in a 10 ml flask, dissolved with 2 ml of methanol and mixed with diluent (blank solution) at a constant volume to obtain.
(62) Ion Chromatograph Test (IC) Conditions (Salt Formation Molar Ratio Test):
(63) TABLE-US-00002 Ion chromatograph ThermoFisher ICS-1100 Chromatographic column IonPac AS18 Analytical Column, 250 * 4 mm Mobile phase 25 mM NaOH Sample injection volume 25 μL Flow rate 1.0 mL/min Temperature/°C 35° C. Column temperature 35° C. Electric current 80 mA Run Time The chloride ion was 6.0 min, the sulfate ion was 8.0 min, the bromide ion was 8.0 min, and the phosphate ion was 12.0 min
Example 1: Preparation and Identification of Crystal Form a of the Free Base of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(64) The free base was prepared according to Example 5 of CN106518841A.
Preparation of 1-benzo[b]thiophene-4-piperazine hydrochloride
(65) ##STR00010##
(66) A mixture of 7.20 g of 7-bromobenzo[b]thiophene, 19.9 g of piperazine anhydride, 4.70 g of sodium tert-butoxide, 0.32 g of (R)-(+)-2,2′-bis (diphenylphosphino)-1,1′-dinaphthyl (BINAP), 0.63 g of dipalladium tris (dibenzylideneacetone) and 150 ml of toluene was refluxed under a nitrogen atmosphere for 1 h. 150 ml of water was poured into the reaction solution, then extracted with ethyl acetate (100 ml×3), washed with water, dried over anhydrous magnesium sulfate, and allowed to evaporate the solvent under reduced pressure (0.01 MPa, 45° C.). The residue was purified by silica gel column chromatography (dichloromethane:methanol:25% aqueous ammonia=100:10:1) to yield 4.60 g of 1-benzo[b]thiophen-4-yl-piperazine in the form of yellow oil. 2 ml of concentrated hydrochloric acid was added to a methanol solution (25 ml) containing 4.6 g of 1-benzo[b]thiophen-4-yl-piperazine, and the solvent was evaporated under reduced pressure (0.01 MPa, 45° C.). Ethyl acetate (50 ml) was added to the residue and the precipitated crystals were filtered, dissolved at reflux in 15 ml of methanol and then cooled to room temperature (25° C.) to recrystallize to obtain 1-benzo[b]thiophen-4-yl-piperazine hydrochloride in the form of colorless needles.
Preparation of tert-Butyl trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl-carbamate
(67) ##STR00011##
(68) 2.54 g (10 mmol) of 1-benzo[b]thiophene-4-piperazine hydrochloride and 2.40 g (10 mmol) of trans-2-{1-[4-(N-tert-butoxycarbonyl) amino]cyclohexyl}-acetaldehyde were dissolved in 120 ml of dichloromethane, 1.40 ml (10 mmol) of triethylamine was added at room temperature (25° C.±2° C.) and stirred slowly for 10 min, then 3.16 g (14.8 mmol) of sodium triacetoxyborohydride was added gradually. The reaction proceeded for a further 24 h at room temperature under stirring, and after reaction, 120 ml of a 10% sodium bicarbonate solution was added. The reaction system was directly subjected to separation, the organic phase was dried over anhydrous sodium sulfate, and finally filtered and concentrated to dryness by rotary evaporation, and the solid was dissolved in 15 ml of ethyl acetate under reflux and cooled to room temperature (25° C.±2° C.) to crystallize to give 3.70 g of the desired product.
Preparation of trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexylamine
(69) ##STR00012##
(70) 4.43 g of tert-Butyl trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl-carbamate was placed in a reaction flask under ice-water bath, 80 ml of saturated hydrogen chloride in ethyl acetate was added, the reaction mixture was stirred for 8 h to carry out deprotection reaction, and finally a white precipitate was formed to give 3.42 g of the hydrochloride salt of the title compound. The above solid was added to 50 ml of dichloromethane solution, followed by addition of 50 ml of saturated sodium bicarbonate solution, stirred for half an hour, then subjected to phase separation. The organic phase was concentrated (0.01 MPa, 40° C.) to give 3.30 g of the desired product.
Preparation of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea
(71) ##STR00013##
(72) 1.73 g of trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexylamine was dissolved in 50 ml of dichloromethane, followed by addition of 1.40 ml of triethylamine and 5.50 mmol of N, N-dimethylcarbamoyl chloride. The mixture was stirred for 48 h at room temperature (25° C.±2° C.). After reaction, 50 ml of water was added for extraction and the organic phase was concentrated (0.01 MPa, 45° C.), subjected to column chromatography (methanol:dichloromethane=1:10, 400 mesh silica gel) to collect the target fraction, which was concentrated to give 1.89 g of the amorphous desired product.
(73) Preparation and Identification of Crystal Form a of the Free Base
(74) 200 mg of the above amorphous product was dissolved in ethyl acetated at reflux temperature of 77° C., cooled to room temperature (20-25° C.) and stirred for 1 h, filtered under suction and recrystallized to obtain a crystal form. It is designated as crystal Form A of the free base of the compound of Formula I, wherein the X-ray diffraction pattern (XRPD) is shown in
Example 2: Preparation and Identification of the Hydrochloride Salt of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(75) Preparation of hydrochloride salt: 200 mg of the free base product from Example 1 was mixed and stirred with 1.05 molar ratio of hydrochloric acid in toluene at room temperature for 3 days and dried under vacuum at 50° C. for 3 days to obtain the hydrochloride salt. It is in crystal form, designated as hydrochloride salt Form B of the compound of Formula I.
(76)
(77) TABLE-US-00003 TABLE 1 XRPD Diffraction Peak Data for Hydrochloride Salt Form B Diffraction Interplanar Relative angle [° 2θ] spacing [Å] intensity [%] 4.576941 19.30686 55.05 9.123195 9.69356 23.68 10.982470 8.05632 76.16 11.903470 7.43497 15.55 12.216890 7.24493 13.80 13.040680 6.78905 47.00 13.738740 6.44563 69.38 15.024150 5.89694 22.99 15.800660 5.60884 100.00 16.914740 5.24185 85.20 17.370510 5.10532 7.84 18.339650 4.83767 73.96 19.119300 4.64212 49.76 19.746390 4.49609 40.83 20.029190 4.43324 32.41 20.682720 4.29462 50.63 21.802960 4.07643 19.48 22.151700 4.01303 20.51 22.947100 3.87570 22.56 23.570630 3.77457 40.16 24.581520 3.62158 15.53 24.984200 3.56412 27.39 25.586650 3.48155 10.42 26.251890 3.39482 15.34 26.533050 3.35948 19.10 27.495840 3.24399 3.76 30.408600 2.93957 7.54 32.725620 2.73655 14.39
(78) For the hydrochloride salt Form B of the compound of Formula I, the solubility in water is greater than 4.6 mg/ml; the DSC results of
Example 3: Preparation and Identification of the Sulfate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(79) Preparation of the sulfate (a): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of sulfuric acid in ethyl acetate (EtOAc) at room temperature for 4 days, and dried under vacuum at 50° C. for 3 days to obtain the sulfate. It is in crystal form, designated as sulfate Form A of the compound of Formula I.
(80)
(81) TABLE-US-00004 TABLE 2 XRPD Diffraction Peak Data for Sulfate Form A Diffraction Interplanar Relative angle [° 2θ] spacing [Å] intensity [%] 6.719282 13.15520 44.09 7.195519 12.28556 10.70 7.947205 11.12514 7.24 9.858717 8.97195 21.50 12.610270 7.01979 3.93 13.414660 6.60062 15.56 14.437150 6.13535 26.09 14.823030 5.97649 13.36 15.240620 5.81366 28.09 15.927590 5.56443 99.41 17.257190 5.13859 31.62 17.781580 4.98822 61.09 18.294180 4.84959 30.33 18.863740 4.70443 53.45 20.187920 4.39875 17.60 20.792550 4.27218 20.94 21.464510 4.13993 100.00 21.711510 4.09339 40.36 22.207430 4.00309 10.20 22.741540 3.91027 12.81 23.204680 3.83326 24.80 23.806940 3.73763 43.59 24.552950 3.62573 7.64 25.532400 3.48883 5.35 26.631630 3.34727 17.27 27.023930 3.29956 21.99 27.515020 3.24178 7.32 28.190910 3.16557 16.24 28.563170 3.12516 14.00 29.829620 2.99530 14.63 32.993550 2.71494 4.90 34.360750 2.60998 5.11 36.462810 2.46419 1.68
(82) For the sulfate Form A of the compound of Formula I, the solubility in water is greater than 8.0 mg/ml; the DSC results of
(83) Preparation of sulfate (b): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of sulfuric acid in isopropylamine (IPA) at room temperature for 4 days, and dried under vacuum at 50° C. for 3 days to obtain the sulfate, which is in crystal form and is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the above sulfate Form A of the compound of Formula I.
(84) Preparation of sulfate (c): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of sulfuric acid in acetonitrile (ACN) at room temperature for 4 days, and dried under vacuum at 50° C. for 3 days to obtain the sulfate, which is in crystal form and is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the above sulfate Form A of the compound of Formula I.
(85) Preparation of sulfate (d): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of sulfuric acid in methylbenzene (toluene) at room temperature for 4 days, and dried under vacuum at 50° C. for 3 days to obtain the sulfate, which is in crystal form and is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the above sulfate Form A of the compound of Formula I.
Example 4: Preparation and Identification of the Phosphate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(86) Preparation of phosphate (a): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of phosphoric acid in ethyl acetate (EtOAc) at room temperature for 5 days to obtain. It is in crystal form, designated as phosphate Form A of the compound of Formula I.
(87)
(88) TABLE-US-00005 TABLE 3 XRPD Diffraction Peak Data for Phosphate Form A Diffraction Interplanar Relative angle [° 2θ] spacing [Å] intensity [%] 6.794465 13.00980 18.09 9.973994 8.86852 17.67 13.986170 6.33215 33.04 15.241080 5.81349 33.97 15.844050 5.59358 100.00 18.154110 4.88669 20.98 19.990390 4.44176 19.30 21.005570 4.22933 30.57 21.241610 4.18287 75.24 21.811130 4.07492 26.04 24.450240 3.64073 8.99 26.019400 3.42462 0.62 27.309390 3.26572 9.19 30.710740 2.91134 3.58 32.056180 2.79215 2.76 35.718340 2.51383 2.08 36.401270 2.46822 2.60
(89) For the phosphate Form A of the compound of Formula I, the solubility in water is greater than 7.6 mg/ml; the DSC results of
(90) Preparation of phosphate (b): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of phosphoric acid in isopropylamine (IPA) at room temperature for 5 days to obtain the phosphate, which is in crystal form and is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the above phosphate Form A of the compound of Formula I.
(91) Preparation of phosphate (c): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of phosphoric acid in acetone at room temperature for 5 days to obtain the phosphate, which is in crystal form and is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the above phosphate Form A of the compound of Formula I.
(92) Preparation of phosphate (d): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of phosphoric acid in acetonitrile (ACN) at room temperature for 5 days to obtain the phosphate, which is in crystal form, which is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the phosphate crystal form A of the compound of Formula I.
(93) Preparation of phosphate (e): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of phosphoric acid in methylbenzene (Toluene) at room temperature for 5 days to obtain the phosphate, which is in crystal form, which is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the phosphate crystal form A of the compound of Formula I.
Example 5: Preparation and Identification of the Maleate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(94) Preparation of maleate (a): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of maleic acid in acetone at room temperature for 5 days to obtain the maleate, which is in a crystal form, designated as maleate Form A of the compound of Formula I.
(95)
(96) TABLE-US-00006 TABLE 4 XRPD Diffraction Peak Data for Maleate Form A Diffraction Interplanar Relative angle [° 2θ] spacing [Å] intensity [%] 7.246772 12.19878 29.23 11.045240 8.01067 8.33 11.804630 7.49700 48.03 12.703730 6.96836 37.24 13.493910 6.56202 40.27 14.495010 6.11099 45.47 15.096910 5.86868 100.00 17.108350 5.18296 38.11 17.567560 5.04850 24.86 18.794690 4.72156 15.60 19.104540 4.64567 36.19 19.655590 4.51666 43.21 20.023940 4.43440 37.51 20.395360 4.35447 13.70 21.030950 4.22429 14.53 21.611370 4.11213 77.73 22.496370 3.95232 10.60 22.997610 3.86730 4.66 24.088590 3.69457 89.87 24.867140 3.58063 13.78 25.336430 3.51536 6.62 26.412080 3.37459 12.01 27.786650 3.21070 4.90 28.292510 3.15443 6.19 28.914740 3.08795 7.44 29.804120 2.99780 4.24 30.770440 2.90583 7.49 31.628070 2.82896 6.05 33.952110 2.64045 1.97
(97) For the maleate Form A of the compound of Formula I, the solubility in water is greater than 2.6 mg/ml; the DSC results of
(98) Preparation of maleate (b): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of maleic acid in isopropylamine (IPA) at room temperature for 5 days to obtain the maleate, which is in crystal form and is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the above maleate Form A of the compound of Formula I.
(99) Preparation of maleate (c): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of phosphoric acid in ethyl acetate (EtOAc) at room temperature for 5 days to obtain the maleate, which is in crystal form and is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the above maleate Form A of the compound of Formula I.
(100) Preparation of maleate (d): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of maleic acid in acetonitrile (ACN) at room temperature for 5 days to obtain the maleate, which is in crystal form and is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the above maleate Form A of the compound of Formula I.
(101) Preparation of maleate (e): 200 mg of the product from Example 1 was mixed and stirred with 1.05 molar ratio of maleic acid in methylbenzene (Toluene) at room temperature for 5 days to obtain, which is in crystal form and is substantially the same as the X-ray diffraction pattern (XRPD), DSC pattern and TGA pattern of the above maleate Form A of the compound of Formula I.
Comparative Example 1: Preparation and Identification of the Tartrate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(102) Preparation of tartrate: 200 mg of the product from Example 1 was mixed and stirred with 1 molar ratio of tartaric acid in acetone at room temperature for 5 days. It is in crystal form, designated as tartrate Form A of the compound of Formula I.
(103) With respect to tartrate Form A of the compound of Formula I,
Comparative Example 2: Preparation and Identification of the Fumarate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(104) Preparation of fumarate: 200 mg of the product from Example 1 was mixed and stirred with 1 molar ratio of fumaric acid in acetone at room temperature for 5 days. It is in crystal form, designated as fumarate Form A of the compound of Formula I.
(105) With respect to fumarate Form A of the compound of Formula I,
Comparative Example 3: Preparation and Identification of the Citrate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(106) Preparation of citrate: 200 mg of the product from Example 1 was mixed and stirred with 1 molar ratio of citric acid in ethyl acetate (EtOAc) at room temperature for 5 days. It is in crystal form, designated as citrate Form A of the compound of Formula I.
(107) With respect to citrate Form A of the compound of Formula I,
Comparative Example 4: Preparation and Identification of the Glycolate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(108) Preparation of glycolate: 200 mg of the product from Example 1 was mixed and stirred with 1 molar ratio of glycollic acid in isopropylamine (IPA) at room temperature for 5 days. It is in crystal form, designated as glycolate Form A of the compound of Formula I.
(109) With respect to glycolate Form A of the compound of Formula I,
Comparative Example 5: Preparation and Identification of the Malate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(110) Preparation of malate: 200 mg of the product from Example 1 was mixed and stirred with 1 molar ratio of L-malic acid in Acetone at room temperature for 5 days. It is in crystal form, designated as malate Form A of the compound of Formula I.
(111) With respect to malate Form A of the compound of Formula I,
Comparative Example 6: Preparation and Identification of the DL-Lactate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(112) Preparation of lactate: 200 mg of the product from Example 1 was mixed and stirred with 1 molar ratio of DL-lactic acid in ethyl acetate (EtOAc) at room temperature for 5 days. It is in crystal form, designated as lactate Form A of the compound of Formula I.
(113) With respect to lactate Form A of the compound of Formula I,
Comparative Example 7: Preparation and Identification of the Succinate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(114) Preparation of succinate: 200 mg of the product from Example 1 was mixed and stirred with 1 molar ratio of succinic acid in ethyl acetate (EtOAc) at room temperature for 5 days. It is in crystal form, designated as succinate Form A of the compound of Formula I.
(115) With respect to succinate Form A of the compound of Formula I,
Comparative Example 8: Preparation and Identification of the Adipate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(116) Preparation of adipate: 200 mg of the product from Example 1 was mixed and stirred with 1 molar ratio of adipic acid in ethyl acetate (EtOAc) at room temperature for 5 days. It is in crystal form, designated as adipate Form A of the compound of Formula I.
(117) With respect to adipate Form A of the compound of Formula I,
Comparative Example 9: Preparation and Identification of the p-Toluenesulfonate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(118) Preparation of p-toluenesulfonate: 200 mg of the product from Example 1 was mixed and stirred with 1 molar ratio of p-toluenesulfonic acid in ethyl acetate (EtOAc) at room temperature for 5 days. It is in crystal form, designated as p-toluenesulfonate Form A of the compound of Formula I. With respect to p-toluenesulfonate Form A of the compound of Formula I,
Comparative Example 10: Preparation and Identification of the Mesylate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(119) Preparation of mesylate: 200 mg of the product from Example 1 were stirred with 1 molar ratio of methylsulfonic acid in isopropylamine (IPA) at room temperature for 5 days. It is in crystal form, designated as mesylate Form A of the compound of Formula I.
(120) With respect to mesylate Form A of the compound of Formula I,
Comparative Example 11: Preparation and Identification of Crystalline Form a of Hydrobromide Salt of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(121) Preparation of hydrobromide salt: 200 mg of the product from Example 1 was mixed and stirred with 1 molar ratio of hydrobromic acid in ethyl acetate (EtOAc) at room temperature for 5 days. It is in crystal form, designated as hydrobromide salt Form A of the compound of Formula I.
(122) With respect to hydrobromide salt Form A of the compound of Formula I,
(123) Through analysis of the XRPD pattern, DSC pattern and TGA pattern of the above salts and crystal forms, based on the higher degree of crystallinity, less TGA weight loss, higher and unique DSC endothermic signal combined with the acid safety level, it can be concluded that the hydrochloride salt, sulfate, phosphate and maleate of the compounds of Formula I are preferred.
Comparative Example 12: Preparation and Identification of the Maleate of N′-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound of Formula I)
(124) Preparation of maleate: the maleate was obtained by adding an anti-solvent to 200 mg of the product from Example 1 in a CHCl.sub.3/IPAc (chloroform/isopropyl acetate) system. It is in crystal form, designated as maleate Form B of the compound of Formula I.
(125)
(126) With respect to maleate Form B of the compound of Formula I, it shows a powder X-ray diffraction pattern (XRPD); the DSC results of
(127) Compared with maleate Form A, maleate Form B can be turned to maleate Form A after being heated, and the stability is lower than that of maleate Form A.
Example 6 Hygroscopicity (DVS) Experiment
(128) Dynamic vapor sorption curves were collected on a DVS Intrinsic by SMS (Surface Measurement Systems).
(129) 20 mg sample of the crystal forms of the Examples and Comparative Examples are placed in the environment of 25° C./80% relative humidity for a dynamic vapor sorption (DVS) test, and the test results determined by HPLC are shown in Table 5.
(130) TABLE-US-00007 TABLE 5 The crystal form Vapor changes after DVS Type of salt sorption Hygroscopicity testing or not Example 1 free base 0.16% Almost no No hygroscopicity Example 2 0.21% Almost no No hydrochloride salt hygroscopicity Example 3 sulfate 0.23% Slight No hygroscopicity Example 4 phosphate 0.61% Slight No hygroscopicity Example 5 maleate 0.12 Almost no No hygroscopicity
(131) The above results show that: within one week, hydrochloride salt, sulfate, phosphate and maleate have almost no hygroscopicity or slight hygroscopicity; Maleate is less hygroscopic than other salts.
Example 7
(132) The in vivo absorption process and characteristics of the free base and four salts thereof (sulfate, hydrochloride salt, phosphate, maleate) of compound (I) in rats were studied. The pharmacokinetics of a single intragastric administration of 1 mg/kg of a compound of Formula I and four salts thereof (calculated by free base concentration in salt form) were studied in SD rats.
Experimental Method
(133) SD rats were given a single intragastric administration of 1 mg/kg free base of Example 1 and four salts thereof (including hydrochloride salt from Example 2, sulfate from Example 3, phosphate from Example 4 and maleate from Example 5). There were 4 male rats in each group. The concentration of the compound of Formula I in plasma was determined, and the pharmacokinetic parameters were calculated according to the concentration-time curve. The results are shown in Table 6.
(134) TABLE-US-00008 TABLE 6 Plasma pharmacokinetic parameters in rats after intragastric administration of 1 mg/kg of the free base of compound of Formula I and four salts thereof (Analysis by non-compartmental model) (Mean ± SD, n = 6) AUC.sub.0-t AUC.sub.0-∞ MRT.sub.0-∞ t.sub.1/2 Tmax Cmax Drug ng .Math. h/mL ng .Math. h/mL h h h ng/mL free base 137 ± 58.9 137 ± 59.0 3.63 ± 0.306 2.78 ± 0.463 0.583 ± 0.289 46.6 ± 13.8 Sulfate 125 ± 28.0 125 ± 28.1 2.87 ± 0.283 2.67 ± 0.267 0.417 ± 0.096 55.2 ± 3.49 Hydrochloride 96.9 ± 29.3 99.1 ± 28.9 4.27 ± 2.20 4.72 ± 2.66 0.625 ± 0.250 36.5 ± 13.1 salt Phosphate 139 ± 37.1 139 + 36.8 3.06 ± 0.434 3.25 ± 1.23 0.833 ± 0.333 51.5 ± 16.4 Maleate 124 ± 5.79 125 ± 7.00 3.07 ± 0.531 4.67 ± 2.17 0.750 ± 0.289 45.4 ± 2.81
(135) As can be seen in Table 6, the maleate has a relatively long half-life, prolongs the time of action in vivo, and is highly bioavailable compared with other salts.
Example 8 In Vivo Individual Difference Analysis of Compound of Formula I and Salts Thereof
(136) 1 Material and Method
(137) 1.1 Drugs
(138) Compound of Formula I and salts thereof (sulfate, hydrochloride salt, phosphate, maleate), supplied by Shanghai Jingxin Biopharmaceutical Co., Ltd.
(139) 1.2 Experimental Animal
(140) Strain: SD rats; Gender: male; Body weight: about 250 grams; Source: shanghai Jiesijie Experimental Animal Co., Ltd
1.3 Experimental Method
1.3.1 Administration Method
(141) Route: single intragastric administration; Capacity: measured in 10 mL/kg body weight
(142) Preparation: weighing appropriate amount of the free base or salt of the compound of Formula I, adding a small amount of 0.5% CMC-Na to grind, adding 0.5% CMC-Na to the volume. The concentration is measured as the base.
(143) 1.3.2 Administration and Sample Collection
(144) 20 SD rats were divided into 5 groups with 4 rats in each group, fasted for 12 h and drank freely before administration. Compound of Formula I or salts was given by intragastric administration at the dose of 1 mg/kg respectively. 100 μL of orbital blood samples were collected at 5 min, 10 min, 20 min, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, 24 h and 36 h before and after administration. The blood was anticoagulated with 1% heparin, the plasma was separated by 8000 rpm centrifugation for 4 min and stored at −40° C. for testing.
(145) 1.4 Plasma Sample Determination Method
(146) LC-MS/MS Method.
(147) 1.5 Data Processing
(148) Pharmacokinetic parameters after administration in rats were calculated using a non-compartmental model of DAS 2.0 software.
(149) 2 Results
(150) The individual and average plasma concentration-time curves of rats after single intragastric administration of 1 mg/kg compound of Formula I or salts thereof are shown in Table 7(a)-(e) and
(151) TABLE-US-00009 TABLE 7a Plasma Concentrations (ng/mL) of Compound of Formula I after intragastric administration of 1 mg/kg Compound of Formula I to rats Time after administration (h) 1♂ 2♂ 3♂ 4♂ Mean ± SD 0 0 0 0 0 0.000 ± 0.000 0.0833 0.419 0.624 12.0 0.543 3.40 ± 5.74 0.167 6.35 11.3 25.8 16.3 14.9 ± 8.30 0.333 26.8 33.0 51.8 57.0 42.2 ± 14.5 0.5 33.6 37.2 51.6 63.3 46.4 ± 13.7 1 29.4 36.3 45.6 63.6 43.7 ± 14.8 2 16.8 19.2 23.9 40.3 25.1 ± 10.6 4 5.38 6.30 7.95 16.0 8.91 ± 4.85 6 2.03 1.94 3.06 8.91 3.99 ± 3.32 9 0.876 0.95 1.47 3.30 1.65 ± 1.13 12 0.372 0.578 0.682 1.66 0.823 ± 0.573 24 0.487 0.308 0.886 0.306 0.497 ± 0.273 Note: part of the 36 h concentration was lower than LLOQ (0.1 ng/mL) and 36 h data are not tabulated.
(152) TABLE-US-00010 TABLE 7b Plasma Concentrations (ng/mL) of Compound of Formula I after intragastric administration of 1 mg/kg the Sulfate of Compound of Formula I to rats Time after administration (h) 1♂ 2♂ 3♂ 4♂ Mean ± SD 0 0 0 0 0 0.000 ± 0.000 0.0833 1.54 2.41 2.56 2.48 2.25 ± 0.476 0.167 24.0 18.4 23.0 21.3 21.7 ± 2.45 0.333 50.4 54.4 51.0 48.9 51.2 ± 2.32 0.5 56.0 36.0 48.0 59.4 49.9 ± 10.4 1 42.6 30.0 38.5 45.8 39.2 ± 6.84 2 32.7 14.5 24.1 21.3 23.2 ± 7.54 4 10.3 4.31 10.5 6.57 7.92 ± 3.01 6 3.08 1.35 3.99 3.05 2.87 ± 1.10 9 1.30 0.643 1.68 0.973 1.15 ± 0.444 12 0.718 0.317 0.827 0.561 0.606 ± 0.221 24 0.213 0 135 0.148 0.479 0.244 ± 0.161 Note: part of the 36 h concentration was lower than LLOQ (0.1 ng/mL) and 36 h data are not tabulated.
(153) TABLE-US-00011 TABLE 7c Plasma Concentrations (ng/mL) of Compound of Formula I after intragastric administration of 1 mg/kg the Hydrochloride Salt of Compound of Formula I to rats Time after administration (h) 1♂ 2♂ 3♂ 4♂ Mean ± SD 0 0 0 0 0 0.000 ± 0.000 0.0833 0.448 1.02 9.68 32.6 10.9 ± 15.0 0.167 5.53 10.3 4.72 25.0 11.4 ± 9.40 0.333 37.6 25.9 8.18 45.9 29.4 ± 16.4 0.5 48.3 30.6 15.0 46.4 35.1 ± 15.6 1 32.4 25.3 20.8 42.7 30.3 ± 9.55 2 19.4 11.5 13.3 24.3 17.1 ± 5.86 4 5.99 3.04 5.43 8.99 5.86 ± 2.45 6 2.26 1.46 2.74 3.90 2.59 ± 1.02 9 0.932 0.683 1.32 1.25 1.05 ± 0.295 12 0.514 0.299 1.74 0.777 0.833 ± 0.636 24 0.139 0.423 0.591 0.190 0.336 ± 0.210 Note: part of the 36 h concentration was lower than LLOQ (0.1 ng/mL) and 36 h data are not tabulated.
(154) TABLE-US-00012 TABLE 7d Plasma Concentrations (ng/mL) of Compound of Formula I after intragastric administration of 1 mg/kg the Phosphate of Compound of Formula I to rats Time after administration (h) 1♂ 2♂ 3♂ 4♂ Mean ± SD 0 0 0 0 0 0.000 ± 0.000 0.0833 5.28 3.26 14.5 3.54 6.65 ± 5.31 0.167 10.5 13.0 14.8 15.0 13.3 ± 2.09 0.333 69.7 25.9 42.5 37.9 44.0 ± 18.5 0.5 66.7 23.2 54.7 37.0 45.4 ± 19.2 1 51.1 34.7 60.6 40.8 46.8 ± 11.4 2 25.9 20.0 37.2 18.2 25.3 ± 8.57 4 10.4 10.7 13.9 6.41 10.4 ± 3.07 6 3.65 3.25 5.23 2.40 3.63 ± 1.19 9 1.45 1.61 1.89 0.978 1.48 ± 0.382 12 0.728 0.799 0.923 0.582 0.758 ± 0.142 24 0.175 0.274 0.216 0.121 0.197 ± 0.0647 Note: part of the 36 h concentration was lower than LLOQ (0.1 ng/mL) and 36 h data are not tabulated.
(155) TABLE-US-00013 TABLE 7e Plasma Concentrations (ng/mL) of Compound of Formula I after intragastric administration of 1 mg/kg the Maleate of Compound of Formula I to rats Time after administration (h) 1♂ 2♂ 3♂ 4♂ Mean ± SD 0 0 0 0 0 0.000 ± 0.000 0.0833 1.15 0.643 0.768 0.624 0.796 ± 0.244 0.167 14.5 17.9 8.05 14.8 13.8 ± 4.14 0.333 41.2 43.9 37.1 39.3 40.4 ± 2.89 0.5 47.4 47.7 43.6 40.6 44.8 ± 3.38 1 45.0 44.1 44.6 41.7 43.9 ± 1.48 2 24.4 21.8 26.7 20.1 23.3 ± 2.90 4 8.41 7.03 7.39 8.13 7.74 ± 0.640 6 3.17 2.38 2.67 2.84 2.77 ± 0.330 9 1.32 1.02 0.998 1.38 1.18 ± 0.199 12 0.835 0.637 0.423 0.985 0.720 ± 0.244 24 0.313 0.101 0.170 0.161 0.186 ± 0.0899 Note: part of the 36 h concentration was lower than LLOQ (0.1 ng/mL) and 36 h data are not tabulated.