Crystal form of lenvatinib mesylate and preparation method therefor

Abstract

A new crystal form of lenvatinib mesylate, a preparation method thereof and a use thereof in the preparation of a drug for treating cancer.

Claims

1. Crystal form X of a mesylate of a compound of formula (I), wherein an X-ray powder diffraction spectrum thereof has characteristic peaks at 2 theta values of 22.84°±0.2°, 20.16°±0.2°, 9.46°±0.2°, 14.52°±0.2°, 24.86°±0.2°, 18.92°±0.2°, 26.82°±0.2°, 8.68°±0.2°, 10.32°±0.2°: ##STR00003##

2. Crystal form X according to claim 1, wherein it has an X-ray powder diffraction spectrum as shown in FIG. 1.

3. Crystal form X according to claim 1, wherein a DSC diagram of the crystal form has characteristic absorption peaks at 245.1° C. and 249.63° C.

4. A preparation method for crystal form X of the mesylate of the compound of formula (I) according to claim 1, wherein the compound of formula (I) and methanesulfonic acid are added into an appropriate crystallization solvent which is a mixed system of water and methanol, and dissolved by heating, which is then cooled down and stirred to give the crystal form.

5. The preparation method according to claim 4, wherein the volume ratio of water to methanol is: 1 mL: (9 mL-10 mL).

6. A method for treating cancer in a subject in need thereof, comprising administering crystal form X of the mesylate of the compound of formula (I) according to claim 1 to the subject.

Description

DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is the XRPD spectrum of crystal form X of lenvatinib mesylate;

(2) FIG. 2 is the DSC diagram of crystal form X of lenvatinib mesylate;

(3) FIG. 3 is the TGA diagram of crystal form X of lenvatinib mesylate;

(4) FIG. 4 is the .sup.1H-NMR spectrum of crystal form X of lenvatinib mesylate;

(5) FIG. 5 is the .sup.13C-NMR spectrum of crystal form X of lenvatinib mesylate;

(6) FIG. 6 is the infrared spectrum of crystal form X of lenvatinib mesylate;

(7) FIG. 7a1 is a comparative XRPD spectrum of the crystal form of lenvatinib mesylate in Example 2 placed for 7 days under a high humidity condition;

(8) FIG. 7a2 is a comparative XRPD spectrum of the crystal form of lenvatinib mesylate in Example 2 placed for 1 month under a high humidity condition;

(9) FIG. 7b is a comparative XRPD spectrum of the crystal form of lenvatinib mesylate in Example 2 placed for 10 days at 2° C. to 8° C.;

(10) FIG. 7c is a comparative XRPD spectrum of the crystal form of lenvatinib mesylate in Example 2 placed for 7 days under a strong light condition;

(11) FIG. 8 is an XRPD spectrum of crystal form X of lenvatinib mesylate in Example 2 at day 0.

EMBODIMENTS OF THE INVENTION

(12) The invention will be further described in detail below in conjunction with specific examples. The examples of the invention are only used to illustrate the technical solutions of the invention, and do not limit the essence and scope of the invention.

(13) The explanations of the abbreviations used in the invention are as follows:

(14) XRPD: X-ray powder diffraction

(15) DSC: Differential Scanning Calorimetry

(16) TGA: Thermogravimetric Analysis

(17) .sup.1H-NMR: Liquid nuclear magnetic hydrogen spectrum

(18) .sup.13C-NMR: Liquid nuclear magnetic carbon spectrum

(19) In the following examples, the test methods were generally implemented under conventional conditions or in accordance with conditions recommended by the manufacturer.

(20) The structure of a compound was determined by nuclear magnetic resonance (.sup.1H NMR). Nuclear magnetic resonance (.sup.1H NMR) shift (δ) was given in the unit of parts per million (ppm); the measurement of nuclear magnetic resonance (.sup.1H NMR) was carried out with Bruker AVANCE-400 nuclear magnetic instrument, with the solvent of hexadeuterated dimethyl sulfoxide (CDCl.sub.3), and the internal standard of tetra methylsilane (TMS).

(21) The X-ray powder diffraction spectrum (XRPD) of the invention was measured by using Liaoning Dandong Haoyuan DX-2700X powder diffractometer. The specific parameters were shown in the following table:

(22) TABLE-US-00002 X-ray diffraction Cu , Kα parameter Kα1:1.540598 ; Kα2:1.544426 Kα2/Kα1 intensity ratio: 0.50 Voltage 40 kilovolt (kV) Current 30 milliampere (mA) Scanning range (2θ°) 3.0 to 40.0 degree Bragg angle (2θ°) 0.02 degree
The differential scanning calorimetry (DSC) measurement data of the invention were collected on TAQ2000 differential scanning calorimeter, and the instrument parameters were shown in the following table:

(23) TABLE-US-00003 DSC Sample pan Aluminum pan, gland Temperature range room temperature to 300° C. Scanning rate 10° C./min Protective gas Nitrogen

(24) The thermogravimetric analysis (TGA) measurement data of the invention were collected on TAQ5000 thermogravimetric analyzer, and the instrument parameters were shown in the following table:

(25) TABLE-US-00004 TGA Sample pan Platinum pan, open Temperature range room temperature to 350° C. Scanning rate 10° C./min Protective gas Nitrogen

(26) “Room temperature” in the invention refers to a temperature between 10° C. and 25° C.

Example 1: Preparation of Crystal Form X of Lenvatinib Mesylate

(27) 20 g of lenvatinib was suspended in a mixed solvent of 180 ml of methanol and 20 ml of water, 4.5 g of methanesulfonic acid (1.0 equivalent) was taken and slowly added to the suspension dropwise, which was stirred at room temperature for 30 minutes, and then heated to 55° C. to dissolve the solid completely. After the solid was completely dissolved, it was filtered while hot. The filtrate was naturally cooled down to room temperature, stirred at room temperature for 20 hours, centrifuged to remove the lower solid, and the obtained solid was dried overnight at a constant temperature of 25° C. The obtained solid was determined as crystal form X of lenvatinib mesylate, with the yield of 71%, and the purity thereof was detected by HPLC as 99.23%. The XRPD spectrum of crystal form X obtained in this example was shown in FIG. 1, .sup.1H-NMR was shown in FIG. 4, .sup.13C-NMR was shown in FIG. 5, and the infrared spectrum was shown in FIG. 6. Wherein, the diffraction angle, interplanar spacing and relative intensity of X-ray powder diffraction were shown in Table 1.

(28) TABLE-US-00005 TABLE 1 Peak Pos. d-spacing Rel. int. No. [°2Th.] [Å] [%] 1 8.68 10.179 30.5 2 9.46 9.338 45.6 3 10.32 8.563 21.9 4 14.58 6.064 41.2 5 17.54 5.053 34.1 6 18.92 4.687 57.8 7 20.16 4.400 79.0 8 20.74 4.280 29.5 9 21.56 4.119 28.6 10 22.24 3.994 50.6 11 22.84 3.890 100.0 12 23.32 3.811 32.5 13 23.82 3.732 41.5 14 24.86 3.579 38.6 15 25.44 3.499 26.6 16 26.82 3.321 43.8

(29) The error of the 2θ diffraction angle was ±0.2.

(30) .sup.1H-NMR data were as follows:

(31) .sup.1H NMR (400 MHz, d.sub.6-DMSO) δ 9.00 (d, J=6.6 Hz, 1H), 8.73 (s, 1H), 8.37 (d, J=9.1 Hz, 1H), 8.10 (s, 1H), 7.96 (d, J=24.3 Hz, 2H), 7.70 (s, 1H), 7.66 (d, J=2.8 Hz, 1H), 7.38 (dd, J=9.1, 2.8 Hz, 1H), 7.29 (d, J=2.4 Hz, 1H), 6.98 (d, J=6.6 Hz, 1H), 4.10 (s, 3H), 2.59 (m, 1H), 2.40 (s, 3H), 0.65-0.70 (m, 2H), 0.57-0.30 (m, 2H).

(32) .sup.13C-NMR data were as follows:

(33) .sup.13C NMR (101 MHz, d.sub.6-DMSO) δ 167.33, 164.82, 161.07, 155.39, 148.13, 146.05, 142.65, 135.76, 128.25, 125.24, 121.98, 121.73, 120.38, 114.05, 103.56, 100.76, 56.92, 40.10, 39.89, 39.74, 39.68, 39.47, 39.26, 39.06, 38.85, 22.34, 6.20.

(34) The infrared data were as follows:

(35) Absorption peaks (cm.sup.−1): 532.78, 549.36, 634, 683.03, 776.43, 810.35, 840.45, 870.92, 910.6, 984.45, 1038, 1088.6, 1153, 1193.4, 1239, 1280, 1349.7, 1395.8, 1454.7, 1526.5, 1588.7, 1687, 1708.6, 3023.4, 3137.7, 3345.8, 3480.4, 3592.

(36) The DSC of crystal form X was as shown in FIG. 2 of the specification.

(37) TGA was as shown in FIG. 3 of the specification.

Example 2: Preparation Method of Crystal Form X of Lenvatinib Mesylate

(38) 2.0 kg of lenvatinib was suspended in a mixed solvent of 18.0 L of methanol and 2.0 L of water, 0.45 kg of methanesulfonic acid (1.0 equivalent) was taken and slowly added to the suspension dropwise (adding dropwise for 2.5 hours), which was stirred at room temperature for 30 minutes, and then heated to 55° C. to dissolve the solid completely (dissolving for about 2.0 hours). After the solid was completely dissolved, it was filtered while hot. The filtrate was naturally cooled down to room temperature, stirred at room temperature for 20 hours, centrifuged to remove the lower solid, and the obtained solid was dried overnight at a constant temperature of 25° C. The obtained solid was determined as crystal form X of lenvatinib mesylate, with the yield of 78%, and the purity thereof was detected by HPLC as 99.14%.

(39) The X-ray powder diffraction data of crystal form X obtained in this example were as shown in Table 2.

(40) TABLE-US-00006 TABLE 2 Peak Pos. d-spacing Rel. int. No. [°2Th.] [Å] [%] 1 8.66 10.205 27.5 2 9.48 9.325 46.9 3 10.28 8.595 19.0 4 14.54 6.086 44.5 5 17.52 5.059 23.7 6 18.82 4.711 38.8 7 20.20 4.393 66.3 8 20.70 4.288 22.6 9 21.48 4.134 20.7 10 22.22 3.998 33.8 11 22.80 3.897 100.0 12 23.32 3.811 13.5 13 23.80 3.736 31.4 14 24.82 3.584 44.0 15 25.36 3.509 19.3 16 26.90 3.312 37.1

(41) The error of the 2θ diffraction angle is ±0.2.

(42) The powder diffraction data were substantially consistent with those in Example 1, as shown in FIG. 8.

Example 3: Preparation Method of Crystal Form X of Lenvatinib Mesylate

(43) 20.0 g of lenvatinib was suspended in a mixed solvent of 200 mL of methanol and 20 mL of water, 4.5 g of methanesulfonic acid (1.0 equivalent) was taken and slowly added to the suspension dropwise, which was stirred at room temperature for 30 minutes, and then heated to 55° C. to dissolve the solid completely. After the solid was completely dissolved, it was filtered while hot. The filtrate was naturally cooled down to room temperature, stirred at room temperature for 20 hours, centrifuged to remove the lower solid, and the obtained solid was dried overnight at a constant temperature of 25° C. The obtained solid was determined as crystal form X of lenvatinib mesylate, with the yield of 73%, and the purity thereof was detected by HPLC as 99.24%.

(44) The X-ray powder diffraction data of crystal form X obtained in this example were substantially consistent with those in FIG. 1.

Test Example 1: Determination of the Dissolution Rate of Crystal Form X of Lenvatinib Mesylate

(45) In order to investigate the difference in the dissolution rate between the sample crystal form X in the examples of the invention and the anhydrous crystal form C disclosed in the patent CN100569753C, the samples of crystal form X obtained in Example 2 and crystal form C were respectively added to 500 ml of medium with a pH of 1.0 (prepared with hydrochloric acid) and 500 ml of medium with a pH of 6.8 (prepared with potassium dihydrogen phosphate buffer solvent), stirred at 37° C. (50 r/min). At 5 min, 10 min, 15 min, 20 min, 30 min, 45 min and 60 min, the concentrations of lenvatinib in the medium were determined by using high performance liquid chromatography. The experimental results were shown in Table 3 below.

(46) TABLE-US-00007 TABLE 3 Time (min) 5 10 15 20 30 45 60 pH Crystal form 1.48 1.54 1.54 1.52 1.56 1.57 1.60 1.0 X (mg/ml) Crystal form 0.41 0.47 0.52 0.55 0.61 0.67 0.72 C (mg/ml) pH Crystal form 0.0036 0.0038 0.0039 0.0039 0.0039 0.0039 0.0040 6.8 X (mg/ml) Crystal form 0.0027 0.0030 0.0031 0.0032 0.0033 0.0034 0.0035 C (mg/ml)

(47) The results of the dissolution rate test indicated that compared with crystal form C of lenvatinib mesylate, crystal form X of the invention showed an excellent dissolution rate and solubility, especially in a medium with a pH of 1.0, the dissolution rate within 5 minutes was 3 times higher than crystal form C.

Test Example 2: Stability Study of Crystal Form X of Lenvatinib Mesylate

(48) 6 samples of crystal form X of lenvatinib mesylate were taken, placed in the clean culture dishes and spread out while keeping open, respectively. a) The stabilities of the samples were investigated under a high humidity (25° C., RH 93.5%) condition, specimens were respectively taken at day 7 and day 30, and XRPD tests were performed on the specimens before and after the placement and compared; b) the stabilities of the samples after 10 days of the placement at a storage temperature of 2-8° C. were investigated, and XRPD tests were performed on the samples before and after the placement and compared; c) the stabilities of the samples after 7 days of the placement under the illumination of the strong light (1.2×10.sup.6 Lux±500 Lx) were investigated, and XRPD tests were performed on the samples before and after the placement and compared. The results were shown in Table 4 below. It can be seen from the results in Table 4 that crystal form X of lenvatinib mesylate of the invention did not undergo crystal transformation before and after the stability tests, and the characteristic peaks maintained a high degree of consistency, demonstrating that crystal form X of lenvatinib mesylate of the invention has quite good stability.

(49) TABLE-US-00008 TABLE 4 Results of the stability tests Result of crystal Condition form X Humidity 93.5% 7 days No obvious change in crystal form 1 month No obvious change in crystal form Temperature 2~8° C. 10 days No obvious change in crystal form illumination 7 days No obvious change in crystal form

Test Example 3: Pharmacokinetic Study on Crystal Form X of Lenvatinib Mesylate in Rats

(50) 1 Test Materials

(51) 1.1 Test Drug

(52) Crystal form X of lenvatinib mesylate, prepared in Example 1, provided by the Synthesis Laboratory of Chengdu Easton Biopharmaceuticals Co., Ltd., yellow solid, batch number: 190301.

(53) 1.2 Test Reagent

(54) Corn oil, provided by Yihai Kerry Food Marketing Co., Ltd., batch number GB19111.

(55) 1.3 Preparation of Test Samples

(56) About 20 mg of the test drug was taken, added into 8 mL of vegetable oil and shaken well, to prepare a 2.5 mg/mL suspension.

(57) 1.4 Test Instruments

(58) 1 mL syringe, ethyl alcohol, cotton, 15 mL centrifuge tube, blood collection tube, EP tube, etc.

(59) 2 Test Animals

(60) 6 SD rats, all males, 200-220 g, provided by Chengdu Dashuo Biotechnology Co., Ltd.

(61) 3 Test Method

(62) The rats were intragastrically administrated with 10 mg/kg of the drug. Before administration and at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h after administration, 0.2 mL of jugular blood was taken and placed in EDTA-K2 anticoagulant tube, centrifuged for 10 min at 3000 r/min. The plasma was separated and frozen in the refrigerator at −80° C. After all blood collections were completed, the blood drug concentration in each blood sample was analyzed and determined autonomously, and the parameters of the test drugs (Tmax, time to maximum plasma concentration; Cmax, maximum plasma concentration; AUC.sub.last) were calculated. Based on the obtained parameters, the average value and standard deviation were calculated, and the detailed results were shown in Table 5.

(63) TABLE-US-00009 TABLE 5 Results of the pharmacokinetic test Test Tmax Cmax AUC.sub.last result (h) (ng/mL) (h * ng/mL) Crystal 1.17 ± 9866.67 ± 90867.23 ± form X 0.41 2565.49 19610.04

(64) It can be seen from Table 5 that the time for crystal form X of the invention to reach the maximum plasma concentration was 1.17±0.41 h, the maximum plasma concentration thereof was 9866.672565.49 ng/mL, and the AUC.sub.last, thereof was 90867.23±19610.04 h*ng/mL, demonstrating that crystal form X has better absorption in vivo in animals, which is beneficial to improve the bioavailability of the drug, thereby enhancing the therapeutic effect of the same.

Test Example 4: The Pharmacokinetic Study on Crystal Form X of Lenvatinib Mesylate in Beagle Dogs

(65) Crystal form X of lenvatinib mesylate (the crystal form obtained in Example 1) was pulverized in a mortar and filled into capsules. Beagle dogs were administered orally (n=4), and then orally administrated with 10 mL of water. The dosage was set to 3 mg/kg by the free form. The dogs were fasted one day before the administration, and started to eat at 8 h after the administration. Blood was collected from the vein before and after the administration, placed in an anticoagulation tube, centrifuged to separate the plasma, and frozen in the refrigerator at −80° C. After all blood collections were completed, the blood drug concentration in each blood sample was analyzed and determined, and the parameters of each test substance (Tmax, time to maximum plasma concentration; Cmax, maximum plasma concentration; AUC.sub.last, etc.) were calculated. Based on the obtained parameters, the average value and standard deviation thereof were calculated. It can be seen from the test results that crystal form X has a better absorption in vivo in animals, which is beneficial to improve the bioavailability of the drug, thereby enhancing the therapeutic effect of the same.