Crystal form of pyrazolyl-amino-pyrimidinyl derivative, and preparation method therefor and use thereof

Abstract

Provided in the present disclosure is crystal form I of compound 1, and a preparation method thereof and the use thereof. ##STR00001##

Claims

1. A crystal form I of compound 1, wherein the crystal form I has an X-ray powder diffraction pattern comprising diffraction peaks at the following positions: 8.600.2, 10.250.2, 11.960.2, 14.350.2, 15.390.2 16.590.2, 17.060.2, and 18.160.2 2, as determined by using CuK radiation; ##STR00005##

2. The crystal form I according to claim 1, wherein the crystal form I meets one or more of the following conditions: (1) the crystal form I has an X-ray powder diffraction pattern further comprising diffraction peaks at one or more of the following positions: 12.770.2, 13.480.2, 14.040.2, 17.270.2, 18.830.2, 20.520.2, 20.770.2, 21.450.2, 22.120.2, 22.790.2, 23.550.2, 24.040.2, 24.400.2, 25.080.2, 25.870.2, 26.510.2, 26.730.2, 26.890.2, 27.360.2, and 28.290.2 2; (2) the crystal form I has a differential scanning calorimetry pattern comprising an endothermic peak at 207.4 C. to 209.2 C.; (3) the crystal form I has a thermogravimetric analysis pattern comprising a weight loss of 0.0% at 30.07 C. to 208.96 C.

3. The crystal form I according to claim 2, wherein the crystal form I has an X-ray powder diffraction pattern further comprising diffraction peaks at one or more of the following positions: 28.930.2, 29.420.2, 30.630.2, 33.000.2, 33.370.2, 34.430.2, and 37.090.2 2.

4. The crystal form I according to claim 2, wherein the crystal form I meets one or more of the following conditions: (1) the crystal form I has an X-ray powder diffraction pattern at 2 comprising diffraction peaks as shown in the table below: TABLE-US-00024 Diffraction d value Relative angle [ 2] [] intensity [%] 8.599 10.27532 94.0 10.250 8.62286 41.5 11.957 7.39559 80.8 12.774 6.92428 14.1 13.482 6.56235 13.1 14.043 6.30159 52.8 14.345 6.16938 33.9 15.392 5.75198 16.1 16.594 5.33787 18.7 17.063 5.19224 56.9 17.270 5.13049 27.3 18.156 4.88225 100.0 18.830 4.70888 6.0 20.523 4.32402 26.0 20.765 4.27428 10.0 21.446 4.14001 43.6 22.118 4.01569 17.0 22.792 3.89842 5.7 23.545 3.77553 10.5 24.043 3.69847 13.5 24.406 3.64422 33.5 25.081 3.54761 20.4 25.868 3.44150 2.8 26.508 3.35979 2.8 26.732 3.33219 5.4 26.889 3.31302 9.2 27.360 3.25704 10.2 28.285 3.15261 6.9 28.930 3.08383 30.2 29.419 3.03366 5.0 30.630 2.91642 3.4 33.000 2.71216 3.7 33.370 2.68298 2.6 34.429 2.60282 2.2 37.089 2.42198 2.0 (2) the crystal form I has a differential scanning calorimetry pattern comprising an endothermic peak at 207.4 C. to 209.2 C., with a heat of fusion of 123.76 J/g; (3) the crystal form I has a thermogravimetric analysis pattern as shown in FIG. 3.

5. The crystal form I according to claim 4, wherein the crystal form I meets one or more of the following conditions: (1) the crystal form I has an X-ray powder diffraction pattern at 2 shown in FIG. 1; (2) the crystal form I has a differential scanning calorimetry pattern shown in FIG. 2.

6. A preparation method for the crystal form I according to claim 1, wherein the preparation method comprises scheme 1 or scheme 2; wherein the scheme 1 comprises the following sequential operations: at 40 C., adding methanol to a solution of the compound 1 and tetrahydrofuran, concentrating the obtained solution, adding methanol again to the concentrated solution, and stirring for crystallization to obtain the crystal form I; ##STR00006## the scheme 2 comprises a step of cooling a solution of the compound 1 in tetrahydrofuran/methanol and isopropanol to obtain the crystal form I.

7. The preparation method for the crystal form I according to claim 6, wherein the preparation method meets one or more of the following conditions: (1) the scheme 1 further comprises the following post-processing steps: filtering, washing, drying under reduced pressure, and sieving to obtain the crystal form I; (2) in the scheme 2, the mass ratio of the compound 1 to tetrahydrofuran/methanol is (1:3) to (1:5); (3) in the scheme 2, regarding the tetrahydrofuran/methanol, the mass ratio of tetrahydrofuran to methanol is (2:1) to (1:2); (4) in the scheme 2, the mass ratio of the compound 1 to isopropanol is (1:11) to (1:13); (5) in the scheme 2, the temperature at which the compound 1 is dissolved in tetrahydrofuran/methanol and isopropanol is 50 C. to 60 C.; (6) in the scheme 2, the cooling lowers the temperature to 0 C. to 5 C.; (7) in the scheme 2, a temperature-holding time after the cooling is 15 to 30 h; and (8) the scheme 2 further comprises the following post-processing steps: filtering, washing, drying under reduced pressure, and sieving to obtain the crystal form I.

8. The preparation method for the crystal form I according to claim 7, wherein the preparation method meets one or more of the following conditions: (1) in the scheme 2, the mass ratio of the compound 1 to tetrahydrofuran/methanol is 1:4; (2) in the scheme 2, regarding the tetrahydrofuran/methanol, the mass ratio of tetrahydrofuran to methanol is 1:1; (3) in the scheme 2, the mass ratio of the compound 1 to isopropanol is 1:12; (4) in the scheme 2, the temperature at which the compound 1 is dissolved in tetrahydrofuran/methanol and isopropanol is 55 C.; (5) in the scheme 2, the cooling lowers the temperature to 0 C.; and (6) in the scheme 2, a temperature-holding time after the cooling is 24 h.

9. The preparation method for the crystal form I according to claim 7, wherein the scheme 1 comprises the following sequential operations: at 40 C., adding methanol to a solution of the compound 1 and tetrahydrofuran, concentrating the obtained solution, adding methanol again to the concentrated solution, and stirring for crystallization to obtain the crystal form I, wherein a temperature at which the compound 1 is dissolved in the tetrahydrofuran to form the solution of the compound 1 and tetrahydrofuran is 50 C. to 60 C.; or, the mass ratio of the compound 1 to the tetrahydrofuran is (1:8) to (1:9); or, the mass ratio of the compound 1 to the methanol added for the first time is (1:16) to (1:18); or, the mass of the concentrated solution is 4 to 6 times of the mass of the compound 1; or, the mass ratio of the compound 1 to the methanol added for the second time is (1:2) to (1:3).

10. The preparation method for the crystal form I according to claim 9, wherein the mass ratio of the compound 1 to the tetrahydrofuran is 1:8.7; or, the mass ratio of the compound 1 to the methanol added for the first time is 1:17; or, the mass of the concentrated solution is 5 times of the mass of the compound 1; or, the mass ratio of the compound 1 to the methanol added for the second time is 1:2.3.

11. The preparation method for the crystal form I according to claim 7, wherein the scheme 2 comprises the following operations: warming the compound 1 and tetrahydrofuran/methanol, adding isopropanol to dissolve the compound 1, adding isopropanol again, and cooling to form a suspension to obtain the crystal form I.

12. A pharmaceutical composition, comprising the crystal form I according to claim 1 and a pharmaceutical excipient.

13. A method for treating or preventing a disease related to JAK kinases in a subject in need thereof, comprising: administering the crystal form I according to claim 1 to the subject, wherein the disease related to JAK kinases is inflammatory bowel disease, psoriasis, vitiligo, atopic dermatitis, systemic lupus erythematosus, asthma, diabetic nephropathy, chronic myelogenous leukemia, essential thrombocythemia, polycythemia vera, myelofibrosis, breast cancer or ovarian cancer.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is an XRPD pattern of the crystal form I.

(2) FIG. 2 is a DSC pattern of the crystal form I.

(3) FIG. 3 is a TGA pattern of the crystal form I.

(4) FIG. 4 is a PLM pattern of the crystal form I.

(5) FIG. 5 is an SEM pattern of the crystal form I.

(6) FIG. 6 is a DVS pattern of the crystal form I.

(7) FIG. 7 is an XRPD pattern of the crystal form I before and after DVS testing.

(8) FIG. 8 is an XRPD pattern of the crystal form II.

(9) FIG. 9 is a DSC pattern of the crystal form II.

(10) FIG. 10 is a TGA pattern of the crystal form II.

(11) FIG. 11 is a PLM pattern of the crystal form II.

(12) FIG. 12 is an XRPD pattern of the crystal form III.

(13) FIG. 13 is a DSC pattern of the crystal form III.

(14) FIG. 14 is a TGA pattern of the crystal form III.

(15) FIG. 15 is a PLM pattern of the crystal form III.

(16) FIG. 16 is an XRPD pattern of the crystal form IV.

(17) FIG. 17 is a DSC pattern of the crystal form IV.

(18) FIG. 18 is a TGA pattern of the crystal form IV.

(19) FIG. 19 is a PLM pattern of the crystal form IV.

(20) FIG. 20 is an XRPD pattern of the crystal form V.

(21) FIG. 21 is a DSC pattern of the crystal form V.

(22) FIG. 22 is a TGA pattern of the crystal form V.

(23) FIG. 23 is a PLM pattern of the crystal form V.

(24) FIG. 24 is an XRPD pattern of the crystal form VI.

(25) FIG. 25 is a DSC pattern of the crystal form VI.

(26) FIG. 26 is a TGA pattern of the crystal form VI.

(27) FIG. 27 is a PLM pattern of the crystal form VI.

(28) FIG. 28 is an XRPD pattern of the amorphous form.

(29) FIG. 29 is a DSC pattern of the amorphous form.

DETAILED DESCRIPTION

(30) The present disclosure is further described below by way of examples; however, the present disclosure is not limited to the scope of the described examples. For the experimental methods in which no specific conditions are specified in the following examples, selections are made according to conventional methods and conditions or according to the product instructions.

(31) XRPD Analysis Method

(32) TABLE-US-00002 1. Test preparation Category Name Remark Instrument Bruker D8 Advance X-ray Or equivalent diffractometer instrument Material Monocrystalline silicon plate Or equivalent plate

(33) TABLE-US-00003 2. Parameter setting Parameter Setting Light pipe Cu:K-Alpha1 ( = 1.54060 ) Generator Voltage 40 KV Current 40 mA Optical path Front Soller 2.5 deg Secondary Soller 2.5 deg Divergence 0.60 mm Mode Fixed Detector Mode Lynxeye (1D mode) PSD development 2.1 angle Scanning Type Coupled Two Theta/Theta parameter Mode Continuous PSD fast Range 4 to 40 deg Step size 0.02 deg Step length 0.12 s

(34) Remark: the above parameters are established according to Bruker D8 XRPD and can be adjusted according to different instruments.

(35) 3. Test and Result Report

(36) An appropriate amount of a sample (e.g., 20 to 50 mg, adjustable) was loaded onto a monocrystalline silicon plate, and uniformly coated on the central area of the monocrystalline silicon plate, as shown in the figure below. If the sample has larger particles, a back-loaded sample plate may be used. When there is no requirement for response intensity, both flat and grooved monocrystalline silicon plates may be used; otherwise, a grooved monocrystalline silicon plate should be used to keep the loading height consistent.

(37) If necessary, a thin layer of vaseline or silicone oil may be coated on the surface of a monocrystalline silicon plate to attach the sample, and the excess sample is gently tapped off. The sample plate was loaded onto the sample holder of the XRPD and scanned, to acquire the pattern, and the results were reported.

(38) DSC Test Method

(39) TABLE-US-00004 1. Test preparation Category Name Backup control Instrument Differential scanning TA Discovery series or calorimeter equivalent instrument Analytical balance Precision: at least 1 mg Material Crucible tray Low-Mass aluminum tray, Tzero or equivalent Cover Aluminium, Tzero or equivalent

(40) TABLE-US-00005 2. Parameter setting Parameter Setting Flow rate of nitrogen 50 ml/min Data acquisition frequency 1.00 s/pt Range of temperature rise Room temperature to 300 C. Rate of temperature rise 10 C./min
3. Test and Result Report

(41) An appropriate amount of a sample (not too full to prevent overflow during heating) was taken and placed in a crucible tray, covered with a cover, and sealed with a cover pressing device. An empty crucible tray was taken as a blank control. The crucible tray and cover used for the blank control should be identical to those used for the sample. The crucible trays were mounted on the corresponding sample holders: the sample crucible tray was placed on the sample holder, and the blank crucible tray was placed on the control holder. A method was selected, and a workstation software was used for data processing. The results were reported.

(42) TGA Test Method

(43) TABLE-US-00006 1. Test preparation Category Name Remark Instrument Thermogravimetric TA TGA55/GA550 or analyzer equivalent instruments Material Sample tray Alumina or platinum material

(44) TABLE-US-00007 2. Parameter setting Parameter Setting Flow rate of purge gas for balance 40 ml/min Flow rate of purge gas for furnace 10 ml/min Range of temperature rise Room temperature to 300 C. Rate of temperature rise 10 C./min
3. Test and Result Report

(45) An empty sample tray was placed in a target position on Auto Sampler, TARE on the workstation was clicked, and then the instrument would automatically weigh the tray, and remove the tare weight after the furnace was closed. About 2 to 10 mg of a sample was weighed precisely and placed in the sample tray with the tare weight removed. The sample information was edited, and a method was selected. Sample analysis was started when Sta was clicked, and a curve of sample weight percentage changing with temperature was automatically recorded by the workstation. Analysis was clicked; Weight change in the drop-down menu was selected; Analyze was clicked to perform analyze command; the weight loss percentage (%) of the sample was automatically calculated by the workstation; and the results were reported.

(46) PLM (Polarized Light Microscopy) Test Method

(47) TABLE-US-00008 1. Test preparation Category Name Instrument Nikon LV100POL polarizing microscope Material Glass slide/cover glass
2. Test and Result Report

(48) Several particles of a sample were placed in mineral oil (e.g., silicone oil) to form a suspended matter, which was then placed on a clean glass slide. An appropriate amount of the suspension was placed on a glass slide and covered with a cover glass. For particles with irregular shapes, the characterization of particle size must also include particle information. The homogeneity of the powder should be checked using appropriate magnification. The results of the microphotographs were reported.

Crystal Form I Preparation and Characterization Example 1

(49) LNK01004 (as an amorphous form, prepared with reference to example 113 of CN 113227074 A) (net content: 1.73 kg, 1.000.02) and tetrahydrofuran (15 kg, 8.7) were added to reactor R1, heated to 50 C. to 60 C., and stirred for 1 to 3 h until completely dissolved.

(50) The temperature was controlled to be 40 C., and methanol (30 kg, 17) was then added. The mixture was concentrated under reduced pressure to 4.0 to 6.0, added with methanol (4 kg, 2.3) again, and stirred for 1 to 3 h.

(51) The suspension was filtered, and the filter cake was washed by adding methanol (2 kg, 1.2). At 40 C. to 50 C., the filter cake was dried under reduced pressure until the moisture and solvent residue were qualified (tetrahydrofuran720 ppm, and methanol5000 ppm). After dried to be qualified, the resulting material was sieved to obtain 1.362 kg of a crystal form I (JR-C200212007-FPF21001) of the final product LNK01004, with an XRPD pattern as shown in FIG. 1, a purity of 99.91%, and a yield of 90%.

(52) TABLE-US-00009 XRPD data of JR-C200212007-FPF21001 Diffraction d value Relative angle [ 20] [] intensity [%] 8.599 10.27532 94.0 10.250 8.62286 41.5 11.957 7.39559 80.8 12.774 6.92428 14.1 13.482 6.56235 13.1 14.043 6.30159 52.8 14.345 6.16938 33.9 15.392 5.75198 16.1 16.594 5.33787 18.7 17.063 5.19224 56.9 17.270 5.13049 27.3 18.156 4.88225 100.0 18.830 4.70888 6.0 20.523 4.32402 26.0 20.765 4.27428 10.0 21.446 4.14001 43.6 22.118 4.01569 17.0 22.792 3.89842 5.7 23.545 3.77553 10.5 24.043 3.69847 13.5 24.406 3.64422 33.5 25.081 3.54761 20.4 25.868 3.44150 2.8 26.508 3.35979 2.8 26.732 3.33219 5.4 26.889 3.31302 9.2 27.360 3.25704 10.2 28.285 3.15261 6.9 28.930 3.08383 30.2 29.419 3.03366 5.0 30.630 2.91642 3.4 33.000 2.71216 3.7 33.370 2.68298 2.6 34.429 2.60282 2.2 37.089 2.42198 2.0
Characterization of Crystal Form I

(53) The acquired typical characterization data of the crystal form I obtained from JR-C200212007-FPF21001 are as shown below. The characterization data show that the crystal form I is a stable solvent-free crystal, which can obtain very high purity, stable melting point, solvent-free encapsulation, and good morphology and particle size distribution, and has no obvious hygroscopicity, and these characteristics are beneficial to the subsequent development and production of bulk API and preparations.

(54) TABLE-US-00010 Characterization data of crystal form I Sample number JR-C200212007-FPF21001 Parameter Method Result Purity HPLC 99.91% Melting point DSC (FIG. 2), 207.4 C. to 209.2 C.; 10 C./min 123.76 J/g X-ray diffraction 3 to 40 (2 theta) High crystallinity Thermal TGA, Weight loss of 0.0% from weight 10 C./min 30.07 C. to 208.96 C. loss (FIG. 3) Solvent residue .sup.1H-NMR None Water content Karl Fisher 0.1% Morphology PLM (FIG. 4) Blocky Morphology SEM (FIG. 5) Blocky Particle size PSD dry method D10 = 6 m; distribution D50 = 14 m; D90 = 31 m Hygroscopicity DVS (FIG. 6) Weight gain of 0.9% under 95% RH No crystal form transformation after completion of DVS characterization (FIG. 7)

Crystal Form I Preparation Example 2

(55) LNK01004 (as an amorphous form, prepared with reference to example 113 of CN 113227074 A) (net content: 5 kg, 1.000.02) and tetrahydrofuran/methanol in 1:1 (20 kg, 4.0) were added to reactor R1 and heated to 50 C. to 60 C.; isopropanol (5 kg, 1.0) was added; and the mixture was stirred for 1 to 3 h until completely dissolved.

(56) Isopropanol (55 kg, 11.0) was added while controlling the temperature at 55 C. for 1 to 3 h.

(57) The reaction temperature was lowered to 0 C. within 5.0 h. The temperature was kept at 0 C. for 24 h to form a suspension.

(58) The suspension was filtered, and the filter cake was washed by adding isopropanol (10 kg, 2.0).

(59) At 40 C. to 50 C., the filter cake was dried under reduced pressure until the moisture and solvent residue were qualified (tetrahydrofuran720 ppm, methanol5000 ppm, and isopropanol5000 ppm). After dried to be qualified, the resulting material was sieved to obtain 4.7 kg of a crystal form I (the characterization data of which were consistent with those of Crystal form I Preparation example 1) of the final product LNK01004, with a purity of 100.0% and a yield of 94%.

Crystal Form I Effect Example 1

(60) 1. Water Activity Experiment at 25 C.

(61) 20 mg of the crystal form I was weighed, added to 1 mL of an acetone/water system having different water activities, and stirred at 25 C. for 10, 12 or 22 days. The resulting solid was filtered and subjected to XRPD characterization.

(62) TABLE-US-00011 Water activity experiment at 25 C. Experiment Result number Solvent Time/day XRPD AW1 Water 10 Crystal form I (a.w. = 1) 22 Crystal form I AW9 Acetone/water 12 Crystal form I (v:v = 36:64) (a.w. = 0.9) AW8 Acetone/water 12 Crystal form I (v:v = 60.4:39.6) (a.w. = 0.8) AW7 Acetone/water 12 Crystal form I (v:v = 75.8:24.2) (a.w. = 0.7) AW2 Acetone/water 10 Crystal form I (91:9, v:v) (a.w. = 0.6) 22 Crystal form I AW3 Acetone/water 10 Crystal form I (94:6, v:v) (a.w. = 0.5) 22 Crystal form I AW4 Acetone/water 10 Crystal form I (97:3, v:v) (a.w. = 0.3) 22 Crystal form I AW5 Acetone/water 10 Crystal form I (99:1, v:v) (a.w. = 0.1) 22 Crystal form I AW6 Acetone 10 Crystal form I (a.w. = 0) 22 Crystal form I

(63) The results of the water activity experiment show that the anhydrous crystal form I stably exists for a long time (up to 22 days) in a wide water activity range (1% to 100%) and is not transformed into a hydrate or other crystal forms, which is beneficial to keeping a stable crystal form after a preparation enters the body, and to having sufficient absorption and exposure after a drug enters the body.

(64) 2. Stability Investigation Results

(65) After the crystal form I was placed under the conditions of high temperature of 60 C. and high humidity of 92.5% RH for 30 days, placed under the condition of illumination of 1ICH (the total illumination was not less than 1.210.sup.6 Lux.Math.hr and the near ultraviolet energy was not less than 200 w.Math.hr/m.sup.2), placed under the accelerated condition (40 C.2 C./75%5% RH) for 6 months, and stored under the long-term condition (25 C.2 C./60%5% RH) for 18 months, the results of appearance, related substances, assay (in terms of water-free and solvent-free basis), moisture, crystal form, content and microbial limit of the crystal form I were not changed.

(66) TABLE-US-00012 Stability experiment Conclusion Placement Investigation on crystal Item condition time Investigation item form Influencing High 60 C. 5 days, Appearance, related Crystal form I factor test temperature 40 C. 10 days, substances, assay (in keeps stable 30 days terms of water-free and solvent-free basis), moisture, crystal form, content High 25 C./92.5% RH 5 days, Appearance, humidity 25 C./75% RH2 10 days, related 30 days substances, assay (in terms of water-free and solvent-free basis), moisture, crystal form, content Illumination 1.2 10.sup.6 Lux .Math. hr, 1 ICH Appearance, related 200 w .Math. hr/m.sup.2 substances, assay (in terms of water-free and solvent-free basis), moisture, crystal form, content Conclusion The influencing factor test data show that: after the crystal form I was placed under the conditions of high temperature of 60 C. and high humidity of 92.5% RH for 30 days and placed under the condition of illumination of 1 ICH (the total illumination was not less than 1.2 10.sup.6 Lux.hr and the near ultraviolet energy was not less than 200 w .Math. hr/m.sup.2), the results of all test items meett he corresponding quality standards; the results showed that the crystal form I drug was stable under the conditions of high temperature, high humidity, and illumination. Accelerated test 40 C. 2 C./ 1 month, Appearance, related Crystal form I 75% 5% RH 2 months, substances, assay keeps stable 3 months (in terms of and water-free and 6 months solvent-free basis), content and moisture; crystal form test added on month 0, month 3 and month 6; and microbial limit test added on month 0 and month 6 Intermediate 30 C. 2 C./ 1 month, Appearance, related condition test 65% 5% RH 3 months, substances, assay (in 6 months, terms of water-free and and solvent-free 9 months basis), content and 12 months moisture; and crystal form and microbial limit tests added on month 0 and month 12 Long term test 25 C. 2 C./ 1 month, Appearance, related 60% 5% RH 3 months, substances, assay (in 6 months, terms of water-free 9 months, and solvent-free 12 months, basis), content and 18 months moisture; crystal and form test added on 24 months month 0, month 3, month 6, month 12 and month 24; and microbial limit test added on month 0, month 12 and month 24 Conclusion After the crystal form I was placed under the accelerated condition (40 C. 2 C./75% 5% RH) for 6 months the long-term condition and under (25 C. 2 C./60% 5% RH) for 18 months, the results of appearance, related substances, assay (in terms of water-free and solvent-free basis), moisture, crystal form I, content and microbial limit were not changed.

Crystal Form I Effect Example 2: Study on Dynamic Solubility of Crystal Form I

(67) About 20 mg of the crystal form I was weighed and placed in a 40 mL glass bottle; 10 mL of a simulated gastrointestinal fluid was added; and the mixture was stirred at 400 rpm at 37 C. About 1 mL of the suspension was taken at 1 h, 4 h and 24 h, respectively, and centrifuged at 37 C., and the solubility of the crystal form I at each time point was determined. After 24 h, the pH of the suspension was determined. The remaining suspension was centrifuged, and the remaining solid was subjected to XRPD characterization.

(68) TABLE-US-00013 Study on dynamic solubility of crystal form I (37 C.) Time Solubility (h) (g/mL) pH after 24 h XRPD 1 97.4 2.1 (simulated Crystal form I 4 92.3 gastric juice) 24 101.6 1 100.5 6.6 (simulated Crystal form I 4 111.0 pre-meal intestinal 24 120.0 fluid) 1 389.4 5.1 (simulated Crystal form I 4 473.7 postprandial 24 627.2 intestinal fluid)

(69) The results show that the crystal form I has good solubility (far greater than 8 g/mL) in the simulated gastrointestinal fluid, and the stable absorption of a preparation can be kept in the subsequent preparation development and production.

Crystal Form I Effect Example 3: Press Testing of Crystal Form I

(70) About 10 mg of the crystal form I (sample number: FR00970-12-SU1) was weighed and pressed by a hydraulic press at a pressure of 10 MPa for 5 min, and the transformation of the crystal form and the change in the crystallinity were studied by XRPD characterization. The results show that the advantageous crystal form I keeps stable at high pressure (10 Mpa), which is beneficial to the subsequent stable production of preparations.

(71) TABLE-US-00014 Study on crystal transformation under pressure Pressure XRPD Remark 10 Mpa Crystal form I The crystal form is not changed, and the crystallinity is not changed

Crystal Form I Effect Example 4: Simulated Dry Grinding of Crystal Form I

(72) About 10 mg of the crystal form I (sample number: FR00970-12-SU1) was weighed and ground for 3 min in a mortar, and the transformation of the crystal form and the change in the crystallinity were studied by XRPD characterization. The results show that the crystal form I keeps stability under the dry grinding condition, which is beneficial to the subsequent production of preparations.

(73) TABLE-US-00015 Simulated dry grinding experiment Investigation method XRPD Remark Dry grinding Crystal The crystal form is not changed, and for 3 min form I the crystallinity is slightly reduced

Crystal Form I Effect Example 5: Simulated Dry Grinding of Crystal Form I

(74) About 10 mg of the crystal form I (sample number: FR00970-12-SU1) was weighed; 40 L of water or ethanol was added; the mixture was ground for 3 min in a mortar; and the transformation of the crystal form and the change in the crystallinity were studied by XRPD characterization. The results show that the advantageous crystal form I keeps stability under the wet grinding condition, which is beneficial to the subsequent production of preparations.

(75) TABLE-US-00016 Simulated wet grinding experiment Solvent XRPD Remark Ethanol Crystal form I The crystal form is not changed, and the crystallinity is slightly reduced Water Crystal form I The crystal form is not changed, and the crystallinity is slightly reduced

Crystal Form I Effect Example 6: Pharmacodynamic Data of Crystal Form I

(76) In this test, a migration detection technology was used to detect the half maximal inhibitory concentrations (IC50) of the crystal form I of compound LNK01004, Ruxolitinib, Tofacitinib and Upadacitinib on activities of JAK1, JAK2, JAK3 and TYK2 kinases. In the test, the initial concentration of the crystal form I of compound LNK01004, Ruxolitinib, Tofacitinib and Upadacitinib for the detection of JAK1, JAK2, JAK3 and TYK2 kinases was 10 M, and dilution was performed according to 3-fold gradient dilution, with a total of 10 concentrations. Duplicate wells were set for detecting, and the ATP concentration was 1 mM. The detection results are as shown in the table below:

(77) TABLE-US-00017 Half maximal inhibitory concentrations (IC50) of the crystal form I on activities of JAK1, JAK2, JAK3, and TYK2 kinases JAK1 JAK2 JAK3 TYK2 IC50 (1 mM (1 mM (1 mM (1 mM (nM) ATP) ATP) ATP) ATP) Crystal form I of 10 <0.51 275 1.0 LNK01004 Ruxolitinib 10 9.9 570 43 Tofacitinib 21 75 67 527 Upadacitinib 0.91 19 202 183

(78) It can be seen from the test results that compared to Ruxolitinib, Tofacitinib and Upadacitinib, the crystal form I of the compound LNK01004 has the stronger inhibitory ability to activities of JAK1, JAK2, JAK3 and TYK2 kinases in the activity test, which shows that the crystal form I of LNK01004 has better pharmaceutical performance at the same concentration and the same dose.

Comparative Example 1 Preparation and Characterization of Crystal Form II

(79) About 50 mg of LNK01004 (as an amorphous form, prepared with reference to example 113 of CN 113227074 A) was weighed and completely dissolved at 50 C. by adding 2 ml of acetone/water (v:v=1:1), and the mixture was filtered with a 0.45 m filter membrane to obtain a clear solution. The resulting clear solution was cooled to 5 C. at a cooling rate of 0.1 C./min. The resulting solid was collected by filtration to obtain the crystal form II. The crystal form II (sample number: FR00970-7-SC12) contained 0.3% acetone residue and had a water content of 12.2%. The crystal form II had a lower dehydration temperature of T.sub.onset 56.8 C., and was a metastable hydrate with high crystallinity.

(80) TABLE-US-00018 Characterization data of crystal form II Crystal form Method Crystal form II Sample number FR00970-7-SC12 Crystallinity XRPD (FIG. 8) High crystallinity (2: 3 to 40) Melting point and DSC (FIG. 9) enthalpy of fusion (10 C./min) Thermal weight TGA (FIG. 10) 13.6% @110 C. loss (10 C./min) Content of .sup.1H-NMR 0.06 equivalents of acetone solvent (DMSO-d6) (Theoretical residual solvent content of 0.3%) Water assay KF 12.2% Morphology PLM (FIG. 11) Columnar

Comparative Example 2 Preparation and Characterization of Crystal Form III

(81) About 50 mg of LNK01004 (as an amorphous form, prepared with reference to example 113 of CN 113227074 A) was weighed and placed into a 2 mL glass bottle; 1 mL of methanol/dichloromethane (v:v=1:1) solvent was added; and the mixture was suspended at 400 rpm at 50 C. for one week. The resulting suspension was filtered, and the resulting solid part was characterized as the crystal form III. The crystal form III (sample number: FR00970-7-SC6) had no residual solvent, had a water content of 6.6%, and was a metastable hydrate with high crystallinity.

(82) TABLE-US-00019 Characterization data of crystal form III Crystal form Method Crystal form III Sample FR00970-7-SC6 number Crystallinity XRPD (FIG. 12) High crystallinity (2: 3 to 40) Melting point DSC (FIG. 13) and enthalpy (10 C./min) of fusion Thermal TGA (FIG. 14) 6.6% @110 C. weight loss (10 C./min) Content of .sup.1H-NMR solvent (DMSO-d6) Water assay KF 6.6% Morphology PLM (FIG. 15) Blocky

Comparative Example 3 Preparation and Characterization of Crystal Form IV

(83) About 50 mg of LNK01004 (as an amorphous form, prepared with reference to example 113 of CN 113227074A) was weighed and placed into a 2 mL glass bottle; 1 mL of tetrahydrofuran/water (v:v=1:1) solvent was added and the mixture was suspended at 400 rpm at 25 C. for one week. The resulting suspension was filtered, and the resulting solid was characterized as the crystal form IV (FIG. 24). The crystal form IV (sample number: FR00970-12-SU3) contained 3% tetrahydrofuran residue and had a water content of 5.4%. After placed in the external environment (20-25 C., 80-95% RH) for 2 days, the crystal form IV was transformed into the crystal form VI. It shows that the crystal form IV is a metastable hydrate with high crystallinity.

(84) TABLE-US-00020 Characterization data of crystal form IV Crystal form Method Crystal form IV Sample FR00970-12-SU3 number Crystallinity XRPD (FIG. 16) High crystallinity (2: 3 to 40) Melting point DSC (FIG. 17) and enthalpy (10 C./min) of fusion Thermal TGA (FIG. 18) 6.6% @110 C. weight loss (10 C./min) Content of .sup.1H-NMR solvent (DMSO-d6) Water assay KF 6.6% Morphology PLM (FIG. 19) Blocky

Comparative Example 4 Preparation and Characterization of Crystal Form V

(85) About 20 mg of LNK01004 (as an amorphous form, prepared with reference to example 113 of CN 113227074 A) was weighed and completely dissolved by adding 1 ml of DMF/n-heptane (v:v=1:1) solvent, and the mixture was filtered with a 0.45 m filter membrane to obtain a clear solution. The resulting clear solution was then placed at room temperature for evaporating slowly to obtain a solid as crystal form V. The crystal form V (sample number: FR00970-11-VD3) contained 1.6 equivalents of DMF. After heated to 150 C. to remove the solvent, the crystal form V was transformed to the crystal form I (the XRPD data of which were consistent with those of Crystal form I Preparation example 1).

(86) TABLE-US-00021 Characterization data of crystal form V Crystal form Method Crystal form V Sample FR00970-11-VD3 number Crystallinity XRPD (FIG. 20) High crystallinity (2: 3 to 40) Melting point DSC (FIG. 21) 205.0 C.; 85 J/g and enthalpy of (10 C./min) fusion Thermal TGA (FIG. 22) 11.6% @140 C. weight loss (10 C./min) Content of .sup.1H-NMR 1.6 equivalents of DMF solvent (DMSO-d6) (Theoretical residual solvent content of 20%) Water assay KF / Morphology PLM (FIG. 23) Blocky /: indicates not performing.

Comparative Example 5 Preparation and Characterization of Crystal Form VI

(87) About 50 mg of LNK01004 (as an amorphous form, prepared with reference to example 113 of CN 113227074 A) was weighed and placed into a 2 mL glass bottle; 1 mL of tetrahydrofuran/water (v:v=1:1) solvent was added and the mixture was suspended at 400 rpm at 25 C. for one week. The resulting suspension was filtered; The resulting solid was characterized as the crystal form IV; After placed in the external environment (20 to 25 C., 80 to 95% RH) for 2 days, the crystal form IV was transformed into the crystal form VI. The crystal form VI (sample number: FR00970-9-TC17) contained 2.3% THF residue and had a water content of 4.8%. The crystal form VI was unstable and transformed into other crystal forms under certain conditions. The crystal form VI was transformed into the crystal form VII when being exposed to 0 humidity, and transformed into the crystal form III when being heated to 120 C. and then cooled to room temperature.

(88) TABLE-US-00022 Characterization data of crystal form VI Crystal form Method Crystal form VI Sample FR00970-9-TC17 number Crystallinity XRPD (FIG. 24) Medium crystallinity (2: 3 to 40) Melting point DSC (FIG. 25) and enthalpy of (10 C./min) fusion Thermal TGA (FIG. 26) 8.2% @95 C. weight loss (10 C./min) Content of .sup.1H-NMR 0.16 equivalents of solvent (DMSO-d6) tetrahydrofuran (Theoretical residual solvent content of 2.3%) Water content KF 4.8% Morphology PLM (FIG. 27) Blocky

Comparative Example 6 Preparation and Characterization of Crystal Form VII

(89) About 50 mg of LNK01004 (as an amorphous form, prepared with reference to example 113 of CN 113227074 A) was weighed and placed into a 2 mL glass bottle; 1 mL of tetrahydrofuran/water (v:v=1:1) solvent was added and the mixture was suspended at 400 rpm at 25 C. for one week. The resulting suspension was filtered and the resulting solid was characterized as the crystal form IV; After equilibrated at 0% RH for 12 h, the crystal form IV was transformed into the crystal form VII. The crystal form VII was stable only at low RH, and the crystal form VII was transformed into the crystal form III within 2 h under a condition of 60% RH. The crystal form VII and the crystal form III had similar XRPD patterns, except that the positions of several peaks were slightly shifted.

Comparative Example 7 Preparation and Characterization of Amorphous Form

(90) About 50 mg of LNK01004 (as an amorphous form, prepared with reference to example 113 of CN 113227074 A) was weighed and placed into a 2 mL glass bottle and completely dissolved at 50 C. by adding 1 mL of acetonitrile/water (v:v=1:1) solvent, and the mixture was filtered with a 0.45 m filter membrane to obtain a clear solution. The resulting clear solution was cooled to 5 C. at a cooling rate of 0.1 C./min. The resulting solid was collected by filtration to obtain an amorphous form (sample number: FR00970-7-SC8). It was found from the crystal form of DSC heating study (heating from 30 C. to melting at a rate of 10 C./min; cooling from melting to 20 C. at a rate of 20 C./min) that for the amorphous form at 30 C. to 110 C., the solvent was removed and for heating to 140 C. to 190 C., the amorphous form was transformed into the crystal form I (the characterization data of which were identical to those of Crystal form I Preparation and characterization example 1).

(91) TABLE-US-00023 Characterization data of crystal form in an amorphous form Crystal form Method Amorphous form Sample FR00970-7-SC8 number Crystallinity XRPD (FIG. 28) Amorphous form (2: 3 to 40) Melting point DSC (FIG. 29) The amorphous form has a glass and enthalpy (10 C./min) transition temperature of 104 C. of fusion Crystal form DSC For the amorphous form at 3 C. to transformation (10 C./min) 110 C., the solvent was removed; by DSC heating At 140 C. to 190 C., the amorphous form was transformed into the crystal form I.

(92) Although the specific embodiments of the present disclosure have been described above, it will be understood by those of skill in the art that these are merely illustrative, and that various alterations or modifications can be made to these embodiments without departing from the principle and essence of the present disclosure. Therefore, the scope of protection of the present disclosure is defined by the appended claims.