SALT OF EGFR INHIBITOR, CRYSTAL FORM, AND PREPARATION METHOD THEREFOR

Abstract

Provided are a salt of an EGFR inhibitor, a crystal form thereof and a preparation method therefor, and an application of the salt and the crystal form in the preparation of a treatment for non-small cell lung cancer. The salt has a structure as shown in formula (II).

##STR00001##

Claims

1. A compound of formula (II), ##STR00008## wherein x is selected from 3.0-5.0.

2. The compound of formula (II) according to claim 1, wherein the compound is in a solid form, the solid form being selected from the group consisting of amorphous form and crystalline form.

3. The compound of formula (II) according to claim 2, wherein the crystalline form has diffraction peaks at the following 2θ of 5.95°, 9.51° and 19.04°in the X-ray powder diffraction pattern thereof.

4. The compound of formula (II) according to claim 19, wherein the X-ray powder diffraction pattern of the crystalline form is shown in FIG. 13.

5. (canceled)

6. The compound of formula (II) according to claim 2, wherein the crystalline form has characteristic diffraction peaks at the following 2θ of 6.39°, 12.39° and 14.18° in the X-ray powder diffraction pattern thereof.

7. The compound of formula (II) according to claim 20, wherein the X-ray powder diffraction pattern of the crystalline form is shown in FIG. 4.

8. (canceled)

9. The compound of formula (II) according to claim 2, wherein the crystalline form has diffraction peaks at the following 2θ of 6.78°, 10.33° and 14.04° in the X-ray powder diffraction pattern thereof.

10. The compound of formula (II) according to claim 21, wherein the X-ray powder diffraction pattern of the crystalline form is shown in FIG. 7.

11. (canceled)

12. The compound of formula (II) according to claim 2, wherein the crystalline form has diffraction peaks at the following 2θ of 7.08°, 10.53° and 12.10° in the X-ray powder diffraction pattern thereof.

13. The compound of formula (II) according to claim 22, wherein the X-ray powder diffraction pattern of the crystalline form is shown in FIG. 10.

14-16. (canceled)

17. A method for treating an EGFR-related disease, comprising administering a therapeutically effective amount of the compound of formula (II) of claim 1, the crystalline form thereof, the crystalline form composition thereof, or the pharmaceutical composition thereof to a patient.

18. The compound of formula (II) according to claim 1, wherein x is 4.0.

19. The compound of formula (II) according to claim 3, wherein the crystalline form has diffraction peaks at the following 2θ of 5.95°, 9.51°, 14.91°, 19.04°, 24.95° and 25.39° in the X-ray powder diffraction pattern thereof.

20. The compound of formula (II) according to claim 6, wherein the crystalline form has diffraction peaks at the following 2θ of 6.39°, 12.39°, 13.03°, 14.18°, 19.03° and 26.76° in the X-ray powder diffraction pattern thereof.

21. The compound of formula (II) according to claim 9, wherein the crystalline form has diffraction peaks at the following 2θ of 6.78°, 10.33°, 14.04°, 18.66°, 20.37° and 25.66° in the X-ray powder diffraction pattern thereof.

22. The compound of formula (II) according to claim 12, wherein the crystalline form has diffraction peaks at the following 2θ of 7.08°, 9.11°, 10.53°, 12.10°, 13.78° and 17.51° in the X-ray powder diffraction pattern thereof.

23. The compound of formula (II) according to claim 1, wherein the compound of formula (II) is in a form of a pharmaceutical composition, comprising a therapeutically effective amount of the compound of formula (II) of claim 1, the crystalline form thereof, or the crystalline form composition thereof, and optionally, a pharmaceutically acceptable carrier, excipient and/or medium.

24. The method according to claim 17, wherein the EGFR-related disease is lung cancer.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0095] FIG. 1 is an XRPD pattern of crystalline form A of the compound of formula (IIA) in Example 2A.

[0096] FIG. 2 is a DSC pattern of crystalline form A of the compound of formula (IIA) in Example 2A.

[0097] FIG. 3 is a TGA pattern of crystalline form A of the compound of formula (IIA) in Example 2A.

[0098] FIG. 4 is an XRPD pattern of crystalline form B of the compound of formula (IIA).

[0099] FIG. 5 is a DSC pattern of crystalline form B of the compound of formula (IIA).

[0100] FIG. 6 is a TGA pattern of crystalline form B of the compound of formula (IIA).

[0101] FIG. 7 is an XRPD pattern of crystalline form C of the compound of formula (IIA).

[0102] FIG. 8 is a DSC pattern of crystalline form C of the compound of formula (IIA).

[0103] FIG. 9 is a TGA pattern of crystalline form C of the compound of formula (IIA).

[0104] FIG. 10 is an XRPD pattern of crystalline form D of the compound of formula (IIA).

[0105] FIG. 11 is a DSC pattern of crystalline form D of the compound of formula (IIA).

[0106] FIG. 12 is a TGA pattern of crystalline form D of the compound of formula (IIA).

[0107] FIG. 13 is an XRPD pattern of crystalline form A of the compound of formula (IIA) in Example 2B.

DETAILED DESCRIPTION

[0108] In order to better understand the content of the present application, further description is given with reference to specific examples, but the specific embodiments are not intended to limit the content of the present application.

Example 1: Preparation of the Compound of Formula (I)

[0109] ##STR00003## ##STR00004## ##STR00005##

[0110] Step 1:

[0111] Compound 1 (500 g, 3.62 mol, 1.0 eq) was dissolved in tetrahydrofuran (3.75 L), methyl magnesium bromide (10.86 mol, 3.62 L, 3.0 eq) was added dropwise to the above solution, and the reaction system was maintained at around 10° C. After the dropwise addition, the reaction system was stirred at 25° C. for 12 h. After the reaction was completed as detected by TLC, the reaction solution was added to a saturated aqueous potassium carbonate solution (1.48 kg of potassium carbonate dissolved in 1.8 L of water), then the resulting mixture was slurried with ethanol and filtered, and the filtrate was collected and concentrated to give compound 2 (crude), which was directly used in the next step. .sup.1HNMR (400 MHz, deuterated dimethyl sulfoxide) 6=7.57-7.54 (m, 0.5H), 6.48-6.40 (m, 0.5H), 1.51 (d, J=4.0 Hz, 3H), 1.48 (d, J=4.0 Hz, 3H).

[0112] Step 2:

[0113] Compound 3 (534 g, 3.49 mol, 1.0 eq) was added to 3.4 L of ethanol, and the reaction solution was added dropwise with an aqueous glyoxal solution (607.12 g, 4.18 mol, 1.2 eq, 40% concentration) at 70° C. to 80° C. and then reacted at 75° C. for 3 h. After the reaction was completed as detected by HPLC, a large amount of solid was precipitated. The solid was filtered, and the filter cake was collected and dried to give compound 4. .sup.1H NMR (400 MHz, deuterated dimethyl sulfoxide) 6=9.18 (s, 2H), 8.93 (d, J=2.4 Hz, 1H), 8.58 (dd, J=2.6, 9.2 Hz, 1H), 8.36 (d, J=9.0 Hz, 1H).

[0114] Step 3:

[0115] Compound 4 (200 g, 1.14 mol, 1.0 eq) was placed in methanol (2 L), and wet Pd/C (15 g) was added slowly under nitrogen atmosphere protection. The reaction system was evacuated and then purged with nitrogen once, and then the reaction system was evacuated and purged with hydrogen 3 times. Hydrogen was introduced to a pressure of 2 MPa. The reaction system was reacted at 20-30° C. for 12 h, and TLC showed completion of the reaction. After the reaction was completed, the reaction was filtered through celite, the filter cake was washed with methanol (1 L), and the filtrates were combined, and concentrated by rotary evaporation at 40-50° C. to precipitate a large amount of brown solid. After filtration under reduced pressure, the filter cake was dried in a vacuum drying oven at 40° C. to constant weight, to give compound 5. .sup.1H NMR (400 MHz, deuterated dimethyl sulfoxide) 6=8.60 (d, J=2.0 Hz, 1H), 8.45 (d, J=2.0 Hz, 1H), 7.73 (d, J=9.0 Hz, 1H), 7.24 (dd, J=2.4, 9.0 Hz, 1H), 6.92 (d, J=2.4 Hz, 1H), 6.07 (s, 2H).

[0116] Step 4:

[0117] Compound 5 (441 g, 3.04 mol, 1.0 eq) was dissolved in dichloromethane (5 L) and methanol (0.5 L), sodium bicarbonate (331.78 g, 3.95 mol, 1.3 eq) was added under nitrogen atmosphere protection, and then iodine chloride (517.90 g, 3.19 mol, 1.05 eq) was slowly added dropwise to the reaction solution, and the reaction system was maintained at around 10° C. After the reaction was completed as monitored by HPLC, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give compound 6. .sup.1H NMR (400 MHz, deuterated dimethyl sulfoxide) 6=8.72 (d, J=2.0 Hz, 1H), 8.51 (d, J=2.0 Hz, 1H), 7.77 (d, J=9.0 Hz, 1H), 7.41 (d, J=9.0 Hz, 1H).

[0118] Step 5:

[0119] Compound 6 (760 g, 2.80 mol, 1.0 eq) and compound 2 (525 g, 3.36 mol, 1.2 eq) were dissolved in 1,4-dioxane (7.6 L) and then palladium acetate (31.47 g, 0.14 mol, 0.05 eq), Xantphos (81.12 g, 0.14 mol, 0.05 eq) and potassium phosphate (892.74 g, 4.21 mol, 1.5 eq) were added. The reaction system was warmed up to 100° C. and stirred under nitrogen atmosphere protection for 12 h, and the reaction was completed as monitored by HPLC. The reaction solution was diluted with dichloromethane (3.8 L) and filtered. The filtrate was collected and concentrated to give a crude product. The crude product was slurried with ethyl acetate to give compound 7. .sup.1H NMR (400 MHz, deuterated dimethyl sulfoxide) 6=8.63 (d, J=2.0 Hz, 1H), 8.49 (d, J=2.2 Hz, 1H), 7.82 (d, J=9.3 Hz, 1H), 7.23 (dd, J=4.2, 9.3 Hz, 1H), 1.88 (d, J=13.9 Hz, 6H).

[0120] Step 6:

[0121] Compound 7 (500 g, 2.23 mol, 1.0 eq) was dissolved in N-methylpyrrolidinone (2.5 L) at room temperature under N.sub.2 protection, 5-bromo-2,4-dichloropyrimidine (1016 g, 4.46 mol, 2.0 eq) and DIEA (466 mL, 2.68 mol, 1.2 eq) were added, and the reaction system was warmed up to 95° C. and stirred for 16 h. After the reaction was completed as monitored by HPLC, the reaction solution was cooled to room temperature. The reaction solution was poured into water, a large amount of solid was precipitated. After filtration, the filter cake was slurried with ethanol (1 L), washed with ethyl acetate (0.5 L), and dried in a vacuum drying oven at 50° C. to constant weight, to give compound 8. .sup.1HNMR (400 MHz, deuterated dimethyl sulfoxide) 6=13.26 (s, 1H), 8.94 (s, 2H), 8.84-8.81 (m, 1H), 8.64 (s, 1H), 8.29 (d, J=9.2 Hz, 1H), 2.09 (s, 3H), 2.06 (s, 3H).

[0122] Step 7:

[0123] Compound 9 (150 g, 0.75 mol, 1.0 eq) and 1-methylpiperazine (100 g, 0.99 mol, 1.33 eq) were added to methanol (1 L), Pd/C (15 g) was added under nitrogen atmosphere protection, and the reaction system was warmed up to 60° C. and stirred under hydrogen pressure (50 Pa) for 48 h. After the reaction was completed as monitored by HPLC, the reaction solution was cooled to 20-30° C. and filtered, the filtrate was concentrated to give a crude product, and the crude product was recrystallized from petroleum ether to give compound 10. .sup.1HNMR (400 MHz, deuterated chloroform) 6=4.13 (s, 2H), 2.70-2.33 (m, 10H), 2.28 (s, 3H), 1.83-1.79 (m, 2H), 1.45 (s, 3H), 1.42-1.39 (m, 3H).

[0124] Step 8:

[0125] Compound 10 (860 g, 3.03 mol, 1.0 eq) was dissolved in methanol (2 L) at room temperature, then hydrochloric acid methanol solution (4 mol, 7.59 L, 10 eq) was added and then the reaction system was stirred at room temperature for 48 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated under reduced pressure to give compound 11. .sup.1H NMR (400 MHz, deuterated water) 6=3.59-3.50 (m, 11H), 3.04 (t, J=13.2 Hz, 2H), 2.94 (s, 3H), 2.36 (d, J=13.6 Hz, 2H), 1.92-1.83 (m, 2H).

[0126] Step 9:

[0127] Compound 11 (1.01 kg, 3.47 mol, 1.0 eq) was added to a solution of dichloromethane (5 L) and methanol (0.5 L) at room temperature, sodium hydroxide (429.86 g, 10.75 mol, 3.1 eq) was added under N.sub.2 atmosphere protection, and the reaction solution was warmed up to 20-30° C. and stirred for 12 h. The reaction solution was filtered, and the filtrate was collected and concentrated to give compound 12. .sup.1H NMR (400 MHz, deuterated chloroform) 6=3.12 (d, J=12.4 Hz, 2H), 2.59-2.35 (m, 12H), 2.47 (s, 3H), 1.82 (d, J=12 Hz, 2H), 1.39-1.33 (m, 2H).

[0128] Step 10:

[0129] 1-chloro-5-fluoro-4-methyl-2-nitrobenzene (1.68 kg, 8.86 mol, 1.0 eq) and compound 12 (1.79 kg, 9.75 mol, 1.1 eq) were dissolved in dimethyl sulfoxide (9 L) at 20° C., and DIEA (1.26 kg, 9.75 mol, 1.1 eq) was added. After the addition was completed, the reaction solution was stirred at 60° C. for 20 h; after the reaction was completed as detected by HPLC, the reaction solution was poured into water (18 L), and then a large amount of yellow solid was precipitated. The reaction system was filtered, the filter cake was slurried with ethanol (3.6 L) and filtered, and the obtained filter cake was dried in vacuum at 40° C. to constant weight, to give compound 13. .sup.1H NMR (400 MHz, deuterated chloroform) 6=7.75 (s, 1H), 6.91 (s, 1H), 3.25-3.22 (m, 2H), 2.67-2.42 (m, 11H), 2.44 (s, 3H), 2.42 (s, 3H), 1.92 (d, J=12 Hz, 2H), 1.65-1.61 (m, 2H).

[0130] Step 11:

[0131] Compound 13 (980.13 g, 2.78 mol, 1.0 eq) and N-methylpyrrolidone (5 L) were added sequentially into a reaction kettle, and the temperature in the reaction kettle was adjusted to 15-20° C. Sodium methoxide (328.15 g, 6.07 mol, 2.2 eq) was added slowly in portions into the reaction kettle, and the reaction system was maintained at 15-25° C. After the reaction was completed as monitored by HPLC, the reaction solution was added into distilled water (20 L) and maintained at 10-20° C., and a large amount of solid was precipitated during adding. The solid was filtered, the filter cake was collected, slurried with ethanol and filtered. The filter cake was collected and dried in a vacuum drying oven to constant weight, to give compound 14. .sup.1H NMR (400 MHz, deuterated methanol) 6=7.77 (s, 1H), 6.73 (s, 1H), 3.94 (s, 3H), 3.04 (m, 2H), 2.80-2.72 (m, 11H), 2.32 (s, 3H), 2.27 (s, 3H), 2.00-2.01 (m, 2H), 1.63-1.62 (m, 2H).

[0132] Step 12:

[0133] Compound 14 (638.01 g, 1.83 mol, 1.0 eq), and methanol (6 L) were added sequentially into a high-pressure reaction kettle, and wet Pd/C (64.01 g) was added under argon atmosphere protection. The reaction system was purged with argon once, purged with hydrogen 3 times, and then subjected to reaction under hydrogen pressure (2 MPa) at a temperature of 20-40° C. After the reaction was completed as monitored by HPLC, the solid was filtered, and the filtrate was collected. The reaction system was then transferred to a 50 L reaction kettle, added with thiourea resin (240.10 g) and stirred at 25° C. for 12 h under nitrogen atmosphere protection. The reaction solution was filtered, and the filtrate was collected. The filtrate was concentrated in a rotary evaporator under reduced pressure to give a solid, the solid was collected and dried in a vacuum drying oven to constant weight, to give compound 15. .sup.1H NMR (400 MHz, deuterated methanol) 6=6.49 (s, 1H), 6.48 (s, 1H), 3.74 (s, 3H), 3.08-2.98 (m, 2H), 2.58-2.42 (m, 11H), 2.24 (s, 3H), 2.09 (s, 3H), 1.85-1.82 (m, 2H), 1.63-1.62 (m, 2H).

[0134] Step 13:

[0135] Compound 15 (910.34 g, 2.91 mol, 1.2 eq), compound 8 (1000.06 g, 2.42 mol, 1.0 eq) and N-methylpyrrolidinone (5 L) were added in order into a reaction kettle. The reaction solution was stirred, added dropwise with methanesulfonic acid (722.14 g, 7.51 mol, 3.1 eq) at 20-40° C., during which the temperature was maintained no higher than 40° C. The reaction solution was warmed up to 85-90° C. After the reaction was completed as monitored by HPLC, sodium hydroxide solution (1 mol, 15 L) was added into the reaction solution, and a large amount of solid was precipitated. The solid was filtered and the filter cake was collected. The filter cake was redissolved in ethanol (10 L), the reaction system was warmed to reflux, the addition of distilled water (20 L) was performed under heating reflux, and a large amount of solid was precipitated. The reaction solution was filtered, and the filter cake was collected. The collected filter cake was added into a reaction kettle, ethanol (15 L) and a solution of hydrochloric acid in ethanol (6 L) (2 L of concentrated hydrochloric acid dissolved in 4 L of ethanol) were added, and then the reaction system was heated for 2 h. Then the reaction system was naturally cooled to room temperature, and a large amount of solid was precipitated. The solid was collected by filtration and dissolved in distilled water (10 L), the solution was added with sodium hydroxide solution (1 mol, 10 L), a large amount of solid was precipitated, followed by stirring for 1 h. The solid was filtered, and the filter cake was collected and dried in a vacuum drying oven to constant weight, to give a compound of formula (I). .sup.1H NMR (400 MHz, deuterated methanol) 6=8.94 (d, J=4 Hz, 1H), 8.92 (1H, br), 8.91 (s, 1H), 8.19 (s, 1H), 7.95 (d, J=9.6 Hz, 1H), 7.59 (s, 1H), 6.72 (s, 1H), 3.84 (s, 3H), 3.16-3.13 (m, 2H), 2.70-2.37 (m, 11H), 2.31 (s, 3H), 2.16 (s, 3H), 2.12 (s, 3H), 2.06 (s, 3H), 2.02-1.99 (m, 2H); 1.71-1.68 (m, 2H).

Example 2A: Preparation of Crystalline Form A of the Compound of Formula (IIA)

[0136] ##STR00006##

[0137] The compound of formula (I) (96 g, 131.16 mmol, 1 eq) was added into ethanol (500 mL) and the suspension was heated up to 78° C. Concentrated hydrochloric acid (1.31 mol, 125.03 mL, 37.5% concentration, 10 eq) was diluted with ethanol (500 mL) and added dropwise to a reaction kettle under reflux, the reaction solution became clear when half amount of the diluted hydrochloric acid was added, and a large amount of solid was precipitated near the end of the addition. The reaction solution was stirred at 75-80° C. for 2 h, and then naturally cooled to 20-30° C., and a large amount of off-white solid was precipitated, followed by stirring for 12 h. The solid was filtered and the filter cake was washed twice with ethanol. The filter cake was collected and dried at 40° C. in a vacuum oven to constant weight.

[0138] The dried compound (500 mg) was weighed and added into 5 mL of ethanol, and the reaction solution was stirred at 40-50° C. for 24 h. The reaction solution was filtered, and the filter cake was collected and dried in a vacuum drying oven at 40° C. to constant weight, to give crystalline form A of the compound of formula (IIA) (an XRPD pattern of crystalline form A is shown in FIG. 1, a DSC pattern thereof in FIG. 2, and a TGA pattern thereof in FIG. 3). .sup.1H NMR (400 MHz, deuterated methanol) 6=9.45 (br s, 1H), 9.04-8.89 (m, 2H), 8.76 (br s, 1H), 8.46 (s, 1H), 8.00 (br d, J=9.0 Hz, 1H), 7.38 (s, 1H), 6.87 (br s, 1H), 4.02-3.41 (m, 13H), 3.29 (br d, J=8.3 Hz, 2H), 3.02-2.67 (m, 5H), 2.27 (br d, J=9.0 Hz, 2H), 2.13 (s, 3H), 2.06 (d, J=14.4 Hz, 8H).

TABLE-US-00006 TABLE 5 XRPD diffraction peaks of crystalline form A of the compound of formula (II) in Example 2A Nos. 2θ (°) Relative intensity (%) 1 6.01 100 2 8.95 6.6 3 9.78 7.3 4 11.93 10.3 5 13.76 8.9 6 15.01 7.8 7 15.50 10.1 8 16.63 4.6 9 17.87 8 10 19.20 19.1 11 19.75 8.1 12 19.79 7.8 13 20.55 7.4 14 20.88 17 15 22.03 9 16 23.86 50.7 17 24.69 15.6 18 25.05 14.2 19 25.50 23.9 20 26.13 8 21 26.66 17.2 22 27.14 26.1 23 27.95 8.1 24 29.54 19.3 25 29.82 26 26 30.53 23.3 27 31.60 14.1 28 31.99 12.7 29 32.60 14.6 30 35.42 15.2 31 35.85 6.8 32 36.27 7.3 33 38.69 10.3

[0139] The DSC curve of crystalline form A prepared in this example has starting points of endothermic peaks at 175.48° C., 232.92° C., and 269.88° C.

[0140] The TGA curve of crystalline form A prepared in this example has a weight loss of 3.439% at 86.3° C., 8.051% at 160.34° C., 12.47% at 219.18° C. and 16.33% at 258.50° C.

Example 2B: Preparation of Crystalline Form A of the Compound of Formula (IIA)

[0141] ##STR00007##

[0142] Tetrahydrofuran (12 L) and purified water (0.8 L) were added into a 50 L reaction kettle, and the reaction solution was heated up to 50-60° C. Then the reaction solution was added with the compound of formula (I) (900 g), stirred until the compound had dissolved, added with activated carbon (80 g), and stirred for 10 min. The reaction solution was filter-pressed to a clean area, the filtrate was cooled to 30-40° C. with stirring for 1 h, further cooled to 10-20° C. with stirring for 6 h and filtered, and the filter cake was dried at 45-55° C. under reduced pressure for 8 h to give a solid (800 g).

[0143] Absolute ethanol (12 L) and the prepared solid were added sequentially into a 50 L reaction kettle, and the reaction solution was heated to 75-78° C. Concentrated hydrochloric acid (36%-38%, 480 mL) diluted with absolute ethanol (4 L) was added dropwise into the reaction kettle. The reaction solution was stirred at 75-78° C. for 2 h, and further stirred at 20-30° C. for 6 h to precipitate a large amount of solid. The solid was filtered, and the filter cake was added with ethanol (5 L) and stirred at 20-30° C. for 0.5 h. The filter cake was collected and dried under reduced pressure at 45-55° C. for 10 h to give crystalline form A of the compound of formula (IIA) (670 g) (an XRPD pattern is shown in FIG. 13).

TABLE-US-00007 TABLE 6 XRPD diffraction peaks of crystalline form A of the compound of formula (II) in Example 2B Nos. 2θ (°) Relative intensity (%) 1 5.95 44.4 2 8.81 11.3 3 9.51 81.4 4 9.65 69.5 5 12.89 19.2 6 13.61 22.0 7 14.91 37.7 8 15.37 12.2 9 15.78 14.1 10 16.48 24.1 11 17.62 15.7 12 19.04 100.0 13 19.36 35.4 14 19.48 16.0 15 20.42 17.6 16 21.69 12.5 17 23.32 10.3 18 23.76 27.2 19 24.60 11.6 20 24.95 44.0 21 25.39 43.6 22 26.01 11.6 23 26.27 12.8 24 27.08 21.2 25 29.45 14.1 26 29.67 10.6 27 30.07 11.9 28 31.50 14.6 29 31.88 21.4

[0144] The crystalline forms obtained in Examples 2A and 2B of the present application have the same or similar characteristic peaks in their XRPD patterns according to the analysis of the angle 20)(°. Therefore, the crystalline forms obtained in Examples 2A and 2B are the same crystalline form.

Example 3: Preparation of Crystalline Form A of the Compound of Formula (IIA)

[0145] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 20 mL glass vial and an appropriate amount of ethanol was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (40° C.) for testing (keeping out of the light), stirred at 40° C. for 1 day and then centrifuged, and the residual solid was placed in a vacuum drying oven (30° C.) for drying overnight (10-16 h) to give the crystalline form A of the compound of formula (IIA). Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 20 mL glass vial and an appropriate amount of isopropanol was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (40° C.) for testing (keeping out of the light), stirred at 40° C. for 1 day and then centrifuged, and the residual solid was placed in a vacuum drying oven (30° C.) for drying overnight (10-16 h) to give the crystalline form A of the compound of formula (IIA).

[0146] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 20 mL glass vial and an appropriate amount of tetrahydrofuran was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (40° C.) for testing (keeping out of the light), stirred at 40° C. for 1 day and then centrifuged, and the residual solid was placed in a vacuum drying oven (30° C.) for drying overnight (10-16 h) to give the crystalline form A of the compound of formula (IIA).

[0147] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 20 mL glass vial and an appropriate amount of acetonitrile was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (40° C.) for testing (keeping out of the light), stirred at 40° C. for 1 day and then centrifuged, and the residual solid was placed in a vacuum drying oven (30° C.) for drying overnight (10-16 h) to give the crystalline form A of the compound of formula (IIA).

[0148] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 20 mL glass vial and an appropriate amount of n-heptane was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (40° C.) for testing (keeping out of the light), stirred at 40° C. for 1 day and then centrifuged, and the residual solid was placed in a vacuum drying oven (30° C.) for drying overnight (10-16 h) to give the crystalline form A of the compound of formula (IIA).

[0149] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 20 mL glass vial and an appropriate amount of ethyl acetate was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (40° C.) for testing (keeping out of the light), stirred at 40° C. for 1 day and then centrifuged, and the residual solid was placed in a vacuum drying oven (30° C.) for drying overnight (10-16 h) to give the crystalline form A of the compound of formula (IIA).

[0150] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 20 mL glass vial and an appropriate amount of acetone was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (40° C.) for testing (keeping out of the light), stirred at 40° C. for 1 day and then centrifuged, and the residual solid was placed in a vacuum drying oven (30° C.) for drying overnight (10-16 h) to give the crystalline form A of the compound of formula (IIA).

Example 4: Preparation of Crystalline Form B of the Compound of Formula (IIA)

[0151] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 20 mL glass vial and an appropriate amount of methanol was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (40° C.) for testing (keeping out of the light), stirred at 40° C. for 1 day and then centrifuged, and the residual solid was placed in a vacuum drying oven (30° C.) for drying overnight (10-16 h) to give the crystalline form B of the compound of formula (IIA).

Example 5: Preparation of Crystalline Form C of the Compound of Formula (IIA)

[0152] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 100 mL glass vial and an appropriate amount of a mixed solvent of methanol and water (methanol:water=10:1 (v/v)) was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (70-80° C.) for testing (keeping out of the light), stirred for 2 h, then cooled to 20-30° C. with stirring overnight, and then centrifuged, and the residual solid was placed in a vacuum drying oven (40° C.) for drying overnight (10-16 h) to give the crystalline form C of the compound of formula (IIA).

[0153] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 100 mL glass vial and an appropriate amount of a mixed solvent of ethanol and water (ethanol:water=2:1 (v/v)) was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (70-80° C.) for testing (keeping out of the light), stirred for 2 h, then cooled to 20-30° C. with stirring overnight, and then centrifuged, and the residual solid was placed in a vacuum drying oven (40° C.) for drying overnight (10-16 h) to give the crystalline form C of the compound of formula (IIA).

[0154] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 100 mL glass vial and an appropriate amount of a mixed solvent of isopropanol and H.sub.2O (isopropanol:H.sub.2O=5:1 (v/v)) was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (70-80° C.) for testing (keeping out of the light), stirred for 2 h, then cooled to 20-30° C. with stirring overnight, and then centrifuged, and the residual solid was placed in a vacuum drying oven (40° C.) for drying overnight (10-16 h) to give the crystalline form C of the compound of formula (IIA).

[0155] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 100 mL glass vial and an appropriate amount of a mixed solvent of isopropanol and water (isopropanol:water=3:1 (v/v)) was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (70-80° C.) for testing (keeping out of the light), stirred for 2 h, then cooled to 20-30° C. with stirring overnight, and then centrifuged, and the residual solid was placed in a vacuum drying oven (40° C.) for drying overnight (10-16 h) to give the crystalline form C of the compound of formula (IIA).

Example 6: Preparation of Crystalline Form D of the Compound of Formula (IIA)

[0156] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 100 mL glass vial and an appropriate amount of a mixed solvent of ethanol and water (ethanol:water=5:1 (v/v)) was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (70-80° C.) for testing (keeping out of the light), stirred for 2 h, then cooled to 20-30° C. with stirring overnight, and then centrifuged, and the residual solid was placed in a vacuum drying oven (40° C.) for drying overnight (10-16 h) to give the crystalline form D of the compound of formula (IIA).

[0157] Crystalline form A of the compound of formula (IIA) (500 mg) was weighed and added into a 100 mL glass vial and an appropriate amount of a mixed solvent of acetone and water (acetone:water=5:1 (v/v)) was added to form a suspension. After a magnetic stir bar was added, the suspension was placed on a magnetic heating stirrer (70-80° C.) for testing (keeping out of the light), stirred for 2 h, then cooled to 20-30° C. with stirring overnight, and then centrifuged, and the residual solid was placed in a vacuum drying oven (40° C.) for drying overnight (10-16 h) to give the crystalline form D of the compound of formula (IIA).

Example 7: Ion Chromatography Detection of Chloride Ions

[0158] Two parts of crystalline form A of the compound of formula (IIA) (about 20 mg) were precisely weighed into two 20 mL volumetric flasks, and dissolved and diluted with a solvent to volume; two parts of the above solutions (1 mL) were precisely transferred into two 20 mL volumetric flasks, respectively, and dissolved and diluted with a solvent to volume, and the solutions were labeled, respectively. The content of chloride ions was then detected by ion chromatograph, and the detection results are shown in Table 7.

TABLE-US-00008 TABLE 7 Weight Chloride ion Average chloride Sample (mg) content (%) ion content (%) Crystalline form A of the 20.77 16.78 16.8 compound of formula (IIA) 20.33 16.75

[0159] It is concluded that after the content of the chloride ions is detected by a chloride ion detector, the number of the chlorine atoms in crystalline form A of the compound (IIA) is about 4 by formula calculations.

Test Example 1: Study on In Vivo Efficacy of the Compound of Formula (I) (1)

[0160] Experimental Method:

[0161] In vivo drug efficacy experiment was performed on xenograft (CDX) BALB/c nude mice subcutaneously implanted with Ba/F.sub.3 (Δ19 del/T790M/C797S) source. BALB/c nude mice (female, 6-8 weeks, weighing about 18-22 g) were bred in SPF-rated environments, and each cage was ventilated individually (5 mice per cage). All animals were free to obtain a standard certified commercial laboratory diet. 48 mice in total were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd for the study. Each mouse was implanted with cells subcutaneously in the right flank for tumor growth. The experiment was started when the mean tumor volume reached about 80-120 mm.sup.3. The test compound was administered orally daily with the compound of formula (I) (5 mg/kg, 15 mg/kg, 45 mg/kg, respectively) administered for 13 consecutive days, and the data are shown in Table 5. Tumor volume was measured twice weekly with a two-dimensional caliper, and the volume was measured in mm.sup.3 and calculated according to the following formula: V=0.5a×b.sup.2, where a and b are the long and short diameters, respectively, of the tumor. Anti-tumor efficacy was determined by dividing the mean tumor increase volume of compound-treated animals by the mean tumor increase volume of untreated animals. The inhibition of tumor growth by the test compound in vivo was evaluated by the tumor volume inhibition value (TGI), wherein the TGI of the group administered with the compound of formula (I) (15 mg/kg) was 74.9%, and the TGI of the group administered with the compound of formula (I) (45 mg/kg) was 101.69%.

[0162] On day 14 after administration to the groups for the efficacy experiment, before the last administration and 2 h after the last administration, the plasma of the mice was collected by blood collection from the submandibular vein, and the plasma of the mice was collected at 1 h, 4 h, 8 h, and 24 h after the administration. About 100 μL of blood was collected each time, placed in an anticoagulation tube, and centrifuged at 8000 rpm for 7 min to collect blood plasma, and the blood plasma was preserved at −80° C. Tissues of mouse lung and tumor were collected at the same time 2 h after the administration and stored at −80° C. Wherein the tumors were divided into two parts (the tumor weight for PD analysis did not exceed 100 mg) for detection and data analysis.

[0163] The experimental results are shown in Tables 8 and 9.

TABLE-US-00009 TABLE 8 Tumor volume (mm.sup.3) Test compound Dosage Day 0 Day 2 Day 5 Day 8 Day 10 Day 12 Day 13 Blank Control N/A 85 143 315 582 765 929 1048 Compound of  5 mg/kg/day 84 108 212 395 505 748 881 formula (I) 15 mg/kg/day 84 76 107 126 176 292 326 45 mg/kg/day 84 56 32 35 42 73 68

TABLE-US-00010 TABLE 9 items Compound of Test Compound formula (I) Dose (mg/kg/day) 15.0 T.sub.1/2 (h) 10.0 AUC.sub.0-last (nM .Math. h) 57037 Plasma (nM), 2 h 3553 Tumor (nmol/kg), 2 h 16667 Lung (nmol/kg), 2 h 32533

Test Example 2: Study on In Vivo Efficacy of the Compound of Formula (I) (2)

[0164] Experimental Method:

1. Cell culture: lung cancer PC-9 cells were cultured in an RPMI 1640 medium containing 10% fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin through in vitro monolayer culture in an incubator at 37° C./5% CO.sub.2. The cells were digested with trypsin-EDTA twice a week for passaging as per conventional practice. At a cell saturation of 80%-90% and a required number, the cells were collected and counted. The cell density was 5×10.sup.6 cells.
2. Cell inoculation: 0.2 mL (containing 5×10.sup.6) of a suspension of PC-9 cells (PBS:Matrigel=1:1) was subcutaneously inoculated in the right back of each mouse, and 64 mice were inoculated in total. On day 7 after inoculation, when mean tumor volume was measured to be 169 mm.sup.3, grouping administration was performed using a randomized stratified grouping method based on tumor volume and mice body weight. PBS is phosphate buffered saline, and Matrigel is matrigel.
3. Administration: the dosage was 50 mg/kg for 0-9 days and 25 mg/kg for 10-21 days; the drug was orally administrated once a day for 3 weeks.

Tumor Measurement and Experimental Indices

[0165] Tumor diameters were measured twice weekly using a vernier caliper. The tumor volume was calculated using the following formula: V=0.5a×b.sup.2, where a and b represent the long diameter and short diameter of the tumor respectively.

[0166] The efficacy of compounds against tumor was evaluated by TGI (%).

[0167] Relative tumor volume (RTV) was calculated based on the results of tumor measurement. The formula is RTV=V.sub.t/V.sub.0, where V.sub.0 represents the tumor volume measured at the time point of grouping and administration (i.e., Do), and V.sub.t represents the tumor volume at a certain measurement for the corresponding mouse, the data of TRTV and CRTV should be measured on the same day.

[0168] TGI (%) reflects the tumor growth inhibition rate. TGI (%)=[(1−(average tumor volume at the end of administration in a treatment group−average tumor volume at the start of administration of the treatment group))/(average tumor volume at the end of treatment of the solvent control group−average tumor volume at the start of treatment of the solvent control group)]×100%.

[0169] Tumor weights will be measured at the end of the experiment and the TGI (%) was calculated.

[0170] Experimental results are shown in Table 10. The TGI of the compound of formula (I) was 100% at day 23.

TABLE-US-00011 TABLE 10 Tumor volume (mm.sup.3) Test compound Dose Day 0 Day 2 Day 6 Day 9 Day 13 Day 16 Day 20 Day 23 Blank Control / 186 257 285 326 482 527 637 921 Compound of 50 mg/kg 185 198 92 75 40 45 76 111 formula (I) (0-9 days) 25 mg/kg (10-21 days)

Test Example 3: Stability Test of Crystalline Form a of the Compound of Formula (II) in Example 2A

[0171] Experimental Instruments:

[0172] Illumination test chamber, model: SHH-100GD-2, conditions: 5000±500 lux (visible light) and 90 mw/cm.sup.2 (ultraviolet).

[0173] Electric heating blast drying oven, model: GZX-9140MBE condition: 60° C.

[0174] Constant temperature and humidity chamber, model: SHH-250SD, condition: 25° C./75% RH.

[0175] Constant temperature and humidity chamber, model: LDS-800Y, condition: 40° C./75% RH and 25° C./60% RH.

[0176] Experimental Method:

[0177] Influencing factor experimental conditions (high temperature of 60° C., high humidity of 75% RH, illumination): an appropriate amount of crystalline form A of the compound of formula (IIA) was placed in a dry and clean open weighing bottle, then the bottle was placed into a dryer under different conditions, and then placed into a corresponding thermostat for observation. The bottle was taken out after being placed for 5 days, 10 days and 30 days, and related substances and contents thereof were measured. Accelerated and long-term experimental conditions (40° C./75% RH, 25° C./60% RH): an appropriate amount of crystalline form A of the compound of formula (IIA) was filled into a double-layer LDPE bags, each layer of LDPE bag was sealed by a binding buckle, then put into an aluminum foil bag for heat sealing, and then put into a corresponding thermostat to observe acceleration and long-term stability.

[0178] Experimental Results:

[0179] The content changes of crystalline form A of the compound of formula (IIA) under the conditions of illumination, high temperature and high humidity are shown in Table 11.

TABLE-US-00012 TABLE 11 Experiments of influencing factors Light stability High humidity Initial experiment High temperature (60° C.) (25° C./75% RH) Items value Day 5 Day 10 Day 5 Day 10 Day 30 Day 5 Total 1.63% 1.99% 2.21% 1.59%  1.63% 1.71% 1.41% impurities Content 97.3% 97.1% 95.1% 99.1% 101.6% 99.4% 99.6% Note: N/A indicates no detection.

[0180] The content changes of crystalline form A of the compound of formula (IIA) under accelerated and long-term conditions are shown in Tables 12 and 13.

TABLE-US-00013 TABLE 12 Results of accelerated experiments (40° C. ± 2° C./75% RH ± 5% RH) Detection Observation time items 0 day 1 month 2 months 3 months 6 months Total 1.63% 1.71% 1.50% 1.56% 1.61% impurities Content 97.3% 99.1% 98.1% 98.6% 97.9%

TABLE-US-00014 TABLE 13 Results of long-time experiments (25° C. ± 2° C./60% RH ± 5% RH) Detection Observation time items 0 day 3 months 6 months 9 months 12 months Total 1.63% 1.61% 1.73% 1.83% 1.69% impurities Content 97.3% .sup. 99% 98.2% 99.4% 97.7%

Test Example 4: Stability Test of Crystalline Form a of the Compound of Formula (II) in Example 2B

[0181] The conditions and the method for the raw material stability test are mainly based on related requirements such as “Guidelines for the Stability Test of APIs and Preparations” (General Principles 9001 of the Four Parts of the Chinese Pharmacopoeia, 2015 Edition) and technical guidelines for stability study on chemical drugs (raw materials and preparations) issued by the National Medical Products Administration in February, 2015 (revised), and are specifically shown in Table 14.

TABLE-US-00015 TABLE 14 Observation Experimental condition time points Sample amount Accelerated 40° C. ± 2° C., Month 1, month 1.0 g/bag, 5 bags test 75% RH ± 5% RH 2, month 3 0.7 g/bag, 5 bags and month 6

[0182] Experimental results are shown in Table 15.

TABLE-US-00016 TABLE 15 Observation Limit Time (month) items requirements 0 1 2 3 6 Sum of ≤3.0% 1.18 1.10 1.18 1.30 1.18 impurities Water (%) ≤7.0% 3.0 3.0 3.5 3.2 2.8