Crystal form and salt form of pyridoimidazole compound and preparation method therefor

Abstract

Disclosed are a crystal form and a salt form of a pyrazolopridine compound, and a preparation method therefor. Further included is the use of the crystal form in preparing anti-influenza virus drugs.

Claims

1. A crystal form A or B of a compound of Formula (I), ##STR00017## wherein n is selected from 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5 and 4, wherein the crystal form A has an X-ray powder diffraction pattern (XRPD) with characteristic diffraction peaks at 2θ angles of 6.61±0.2°, 9.27±0.2°, 14.66±0.2°, wherein the crystal form B has an X-ray powder diffraction pattern (XRPD) with characteristic diffraction peaks at 2θ angles of 7.14±0.2°, 11.19±0.2°, 22.39±0.2°.

2. The crystal form A or B of the compound of Formula (I) according to claim 1, wherein the crystal form A of the compound of Formula (I) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 6.61±0.2°, 9.27±0.2°, 14.66±0.2°, 16.69±0.2°, 18.65±0.2°, 19.79±0.2°, 21.85±0.2°, 24.63±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00023 2θ No. (±0.2°) 1 4.693 2 6.606 3 7.371 4 9.272 5 10.396 6 14.66 7 16.219 8 16.693 9 17.502 10 18.648 11 19.099 12 19.793 13 20.683 14 21.846 15 22.814 16 23.188 17 23.642 18 24.631 19 24.964 20 25.516 21 26.385 22 27.138 23 27.946 24 29.426 25 30.236 26 31.204 27 31.675 28 33.02 29 33.65 30 35.623 31 36.259 32 38.665 further, the crystal form A of the compound of Formula (I) has an XRPD pattern as shown in FIG. 1; further, the crystal form A of the compound of Formula (I) has a differential scanning calorimetry (DSC) curve with a starting point of an endothermic peak at 185.46° C.±3° C., and further has a DSC pattern as shown in FIG. 2; further, the crystal form A of the compound of Formula (I) has a thermogravimetric analysis (TGA) curve with a weight loss of 2.479% at 120.00° C.±3° C., and further has a TGA pattern as shown in FIG. 3; further, for the crystal form A of the compound of Formula (I), the compound of Formula (I) has a structure as represented by Compound 1: ##STR00018##

3. The crystal form A or B of the compound of Formula (I) according to claim 1, wherein the crystal form B has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 7.14±0.2°, 11.19±0.2°, 12.00±0.2°, 17.28±0.2°, 18.84±0.2°, 22.39±0.2°, 26.90±0.2°, 27.95±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00024 2θ No. (±0.2°) 1 7.143 2 8.662 3 11.192 4 12.003 5 14.076 6 14.512 7 15.222 8 15.912 9 16.546 10 17.276 11 18.088 12 18.837 13 19.55 14 19.964 15 20.536 16 21.166 17 22.393 18 22.808 19 23.658 20 24.032 21 25.037 22 25.497 23 25.871 24 26.562 25 26.898 26 27.946 27 29.566 28 30.181 29 30.889 30 31.759 31 32.294 32 32.687 33 33.4 34 34.246 35 34.721 36 36.225 37 38 further, the crystal form B of the compound of Formula (I) has an XRPD pattern as shown in FIG. 4; further, the crystal form B of the compound of Formula (I) has a differential scanning calorimetry (DSC) curve with an endothermic peak at 101.04° C.±3° C., and a starting point of an endothermic peak at 188.30° C.±3° C., and further has a DSC pattern as shown in FIG. 5; further, the crystal form B of the compound of Formula (I) has a thermogravimetric analysis (TGA) curve with a weight loss of 4.087% at 154.18° C.±3° C. and a weight loss of up to 4.610% at 196.80° C.±3° C., and further has a TGA pattern as shown in FIG. 6; further, for the crystal form B of the compound of Formula (I), the compound of Formula (I) has a structure as represented by Compound 2: ##STR00019##

4. A compound of Formula (II) as represented by the following formula, or a crystal form C or D thereof: ##STR00020## wherein, n.sub.2 is selected from 1; m.sub.2 is selected from 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5 and 4; the crystal form C of the compound of Formula (II) has an X-ray powder diffraction (XRPD) pattern with characteristic diffraction peaks at 2θ angles of 8.00±0.2°, 15.06±0.2°, 15.84±0.2°; the crystal form D of the compound of Formula (II) has an X-ray powder diffraction (XRPD) pattern with characteristic diffraction peaks at 2θ angles of 6.96±0.2°, 10.31±0.2°, 14.95±0.2°.

5. The compound of Formula (II) as represented by the following formula, or the crystal form C or D thereof according to claim 4, wherein the crystal form C of the compound of Formula (II) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 5.90±0.2°, 6.52±0.2°, 8.00±0.2°, 12.28±0.2°, 15.06±0.2°, 15.84±0.2°, 21.22±0.2°, 26.82±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00025 2θ No. (±0.2°) 1 5.896 2 6.525 3 7.65 4 8.004 5 10.647 6 11.615 7 12.285 8 12.834 9 13.977 10 15.064 11 15.837 12 17.036 13 17.888 14 19.762 15 21.218 16 21.871 17 24.566 18 25.44 19 26.031 20 26.822 21 31.148 22 32.943 23 37.924 24 38.066 further, the crystal form C of the compound of Formula (II) has an XRPD pattern as shown in FIG. 7; further, the crystal form C of the compound of Formula (II) has a differential scanning calorimetry (DSC) curve with an endothermic peak at 193.754° C.±3° C. and an endothermic peak at 235.53° C.±3° C., and further has a DSC pattern as shown in FIG. 8; further, the crystal form C of the compound of Formula (II) has a thermogravimetric analysis (TGA) curve with a weight loss of 5.000% at 117.79° C.±3° C., and a weight loss of up to 12.377% at 222.15° C.±3° C., and further has a TGA pattern as shown in FIG. 9; further, for the crystal form C of the compound of Formula (II), the compound of Formula (II) has a structure as represented by Compound II-1: ##STR00021##

6. The compound of Formula (II), or the crystal form C or D thereof according to claim 4, wherein the crystal form D of the compound of Formula (II) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 6.96±0.2°, 9.44±0.2°, 10.31±0.2°, 14.95±0.2°, 17.38±0.2°, 20.67±0.2°, 21.89±0.2°, 22.72±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00026 No. 2θ (±0.2°) 1 6.958 2 9.444 3 10.311 4 12.128 5 12.819 6 14.946 7 15.322 8 15.892 9 16.268 10 17.376 11 18.698 12 19.72 13 20.666 14 21.89 15 22.717 16 23.806 17 24.63 18 24.907 19 25.792 20 26.704 21 27.452 22 28.202 23 28.595 24 28.966 25 30.843 26 31.198 27 31.75 28 32.584 29 34.359 30 35.168 31 35.816 32 37.196 33 37.569 further, the crystal form D of the compound of Formula (II) has an XRPD pattern as shown in FIG. 10; further, the crystal form D of the compound of Formula (II) has a differential scanning calorimetry (DSC) curve with an endothermic peak at 193.68° C.±3° C., and further has a DSC pattern as shown in FIG. 11; further, the crystal form D of the compound of Formula (II) has a thermogravimetric analysis (TGA) curve with a weight loss of 0.231% at 78.99° C.±3° C. and a weight loss of up to 5.826% at 198.74° C.±3° C., and further has a TGA pattern as shown in FIG. 12; further, for the crystal form D of the compound of Formula (II), the compound of Formula (II) is represented by Compound II-2: ##STR00022##

7. A Compound 3 represented by the following formula, or a crystal form E thereof, ##STR00023## wherein the crystal form E of Compound 3 has an X-ray powder diffraction (XRPD) pattern with characteristic diffraction peaks at 2θ angles of 8.10±0.2°, 9.60±0.2°, 22.97±0.2°.

8. The Compound 3 or the crystal form E thereof according to claim 7, wherein the crystal form E of Compound 3 has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 8.10±0.2°, 9.60±0.2°, 16.09±0.2°, 17.61±0.2°, 18.42±0.2°, 22.97±0.2°, 23.58±0.2°, 25.14±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00027 No. 2θ (±0.2°) 1 8.104 2 9.599 3 9.833 4 11.066 5 11.712 6 12.487 7 12.978 8 13.707 9 15.753 10 16.093 11 16.722 12 16.996 13 17.612 14 18.415 15 19.227 16 19.542 17 20.077 18 20.351 19 21.513 20 22.204 21 22.595 22 22.974 23 23.254 24 23.585 25 24.199 26 24.907 27 25.143 28 25.595 29 26.073 30 26.765 31 27.252 32 27.507 33 27.885 34 28.417 35 28.932 36 29.564 37 30.076 38 31.055 39 31.673 40 32.263 41 32.539 42 33.116 43 34.414 44 35.364 45 35.634 46 35.735 47 37.236 48 37.851 49 38.124 50 38.953 further, the crystal form E of Compound 3 has an XRPD pattern as shown in FIG. 13; further, the crystal form E of Compound 3 has a differential scanning calorimetry (DSC) curve with a starting point of an endothermic peak at 258.27° C.±3° C., and further has a DSC pattern as shown in FIG. 14; further, the crystal form E of Compound 3 has a thermogravimetric analysis (TGA) curve with a weight loss of 0.905% at 121.35° C.±3° C., and further has a TGA pattern as shown in FIG. 15.

9. A compound of Formula (III) as represented by the following formula, or a crystal form F or G thereof: ##STR00024## wherein, n.sub.3 is selected from 1; m.sub.3 is selected from 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5 and 4; wherein the crystal form F of the compound of Formula (III) has an X-ray powder diffraction (XRPD) pattern with characteristic diffraction peaks at 2θ angles of 6.47±0.2°, 9.11±0.2°, 9.90±0.2°; wherein the crystal form G of the compound of Formula (III) has an X-ray powder diffraction (XRPD) pattern with characteristic diffraction peaks at 2θ angles of 6.23±0.2°, 7.20±0.2°, 14.30±0.2°.

10. The compound of Formula (III), or the crystal form F or G thereof according to claim 9, wherein the crystal form F of the compound of Formula (III) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 6.47±0.2°, 9.11±0.2°, 9.90±0.2°, 15.85±0.2°, 16.28±0.2°, 19.40±0.2°, 20.37±0.2°, 24.10±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00028 No. 2θ (±0.2°) 1 4.477 2 6.467 3 9.109 4 9.895 5 11.189 6 11.779 7 12.899 8 14.473 9 15.34 10 15.854 11 16.285 12 17.416 13 17.885 14 18.693 15 19.402 16 20.374 17 21.377 18 22.221 19 23.235 20 23.59 21 24.099 22 24.707 23 26.368 24 27.608 25 28.159 26 29.862 27 31.77 28 32.327 29 35.334 30 37.497 further the crystal form F of the compound of Formula (III) has an XRPD pattern as shown in FIG. 16; further, the crystal form F of the compound of Formula (III) has a differential scanning calorimetry (DSC) curve with an endothermic peak at 78.73° C.±3° C., a starting point of an endothermic peak at 222.37° C.±3° C., and an exothermic peak at 245.01° C.±3° C., and further has a DSC pattern as shown in FIG. 17; further, the crystal form F of the compound of Formula (III) has a thermogravimetric analysis (TGA) curve with a weight loss of 1.192% at 39.57° C.±3° C., a weight loss of up to 3.683% at 81.27° C.±3° C. and a weight loss of up to 6.023% at 199.63° C.±3° C., and further has a TGA pattern as shown in FIG. 18; further, for the crystal form F of the compound of Formula (III), the compound of Formula (III) is represented by Compound III-1: ##STR00025##

11. The compound of Formula (III), or the crystal form F or G thereof according to claim 9, wherein the crystal form G of the compound of Formula (III) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 6.23±0.2°, 7.20±0.2°, 7.81±0.2°, 11.22±0.2°, 12.38±0.2°, 14.30±0.2°, 15.90±0.2°, 18.97±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00029 No. 2θ (±0.2°) 1 4.806 2 5.933 3 6.23 4 6.527 5 7.197 6 7.809 7 9.5 8 9.9 9 10.182 10 11.217 11 11.785 12 12.383 13 12.915 14 13.327 15 13.918 16 14.298 17 14.634 18 15.617 19 15.895 20 16.439 21 16.737 22 17.071 23 17.555 24 17.931 25 18.177 26 18.969 27 19.921 28 20.173 29 21.065 30 21.984 31 22.401 32 22.679 33 22.816 34 23.528 35 25.319 36 26.107 37 27.315 38 28.063 39 28.753 40 30.273 41 30.905 42 31.454 43 33.08 44 33.43 45 35.019 46 35.402 47 36.172 48 36.721 49 37.749 50 38.508 further, the crystal form G of the compound of Formula (III) has an XRPD pattern as shown in FIG. 19; further, the crystal form G of the compound of Formula (III) has a differential scanning calorimetry (DSC) curve with an endothermic peak at 70.13° C.±3° C., and further has a DSC pattern as shown in FIG. 20; further, the crystal form G of the compound of Formula (III) has a thermogravimetric analysis (TGA) curve as shown in FIG. 21; further, the crystal form G of the compound of Formula (III), the compound of Formula (III) is represented by Compound III-1: ##STR00026##

12. A compound of Formula (IV) as represented by the following formula, or a crystal form H, K, I or J thereof: ##STR00027## wherein, n.sub.4 is selected from 1; m.sub.4 is selected from 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5 and 4; wherein the crystal form H of the compound of Formula (IV) has an X-ray powder diffraction (XRPD) pattern with characteristic diffraction peaks at 2θ angles of 4.71±0.2°, 5.56±0.2°, 18.16±0.2°; wherein the crystal form K of the compound of Formula (IV) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 4.83±0.2°, 7.39±0.2°, 14.80±0.2°; wherein the crystal form I of the compound of Formula (IV) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 4.89±0.2°, 6.19±0.2°, 7.45±0.2°; wherein the crystal form J of the compound of Formula (IV) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 4.97±0.2°, 16.33±0.2°, 23.92±0.2°.

13. The compound of Formula (IV), or the crystal form H, K, I or J thereof according to claim 12, wherein the crystal form H of the compound of Formula (IV) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 4.71±0.2°, 5.56±0.2°, 7.98±0.2°, 8.97±0.2°, 18.16±0.2°, 22.42±0.2°, 26.37±0.2°, 27.10±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00030 No. 2θ (±0.2°) 1 4.71 2 5.559 3 7.984 4 8.971 5 12.543 6 13.361 7 14.352 8 15.186 9 16.125 10 16.743 11 18.163 12 18.518 13 19.151 14 19.659 15 19.921 16 22.42 17 23.268 18 26.367 19 27.097 20 27.574 21 28.355 22 29.319 23 30.035 24 32.92 further, the crystal form H of the compound of Formula (IV) has an XRPD pattern as shown in FIG. 22; further, the crystal form H of the compound of Formula (IV) has a differential scanning calorimetry (DSC) curve with an endothermic peak at 141.17° C.±3° C., an endothermic peak at 243.06° C.±3° C., and an exothermic peak at 257.74° C.±3° C., and further has a DSC pattern as shown in FIG. 23; further, the crystal form H of the compound of Formula (IV) has a thermogravimetric analysis (TGA) curve with a weight loss of 1.328% at 73.74±3° C., a weight loss of up to 4.986% at 207.43° C.±3° C., and a weight loss of up to 5.627% at 249.40° C.±3° C., and further has a TGA pattern as shown in FIG. 24.

14. The compound of Formula (IV), or the crystal form H, K, I or J thereof according to claim 12, wherein the crystal form K of the compound of Formula (IV) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 4.83±0.2°, 7.39±0.2°, 11.61±0.2°, 14.81±0.2°, 16.19±0.2°, 18.50±0.2°, 19.29±0.2°, 20.86±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00031 No. 2θ (±0.2°) 1 4.827 2 5.989 3 7.393 4 9.814 5 11.14 6 11.612 7 14.204 8 14.473 9 14.807 10 15.042 11 16.187 12 18.499 13 18.933 14 19.287 15 19.66 16 20.863 17 22.48 18 23.292 19 26.245 20 26.504 21 27.841 22 28.477 23 34.215 further, the crystal form K of the compound of Formula (IV) has an XRPD pattern as shown in FIG. 31; further, the crystal form K of the compound of Formula (IV) has a differential scanning calorimetry (DSC) curve as shown in FIG. 32; further, the crystal form K of the compound of Formula (IV) has a thermogravimetric analysis (TGA) curve with a weight loss of 3.442% at 83.69° C.±3° C. and a weight loss of up to 4.947% at 183.76° C.±3° C., and further has a TGA pattern as shown in FIG. 33.

15. The compound of Formula (IV), or the crystal form H, K, I or J thereof according to claim 12, wherein the crystal form I of the compound of Formula (IV) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 4.89±0.2°, 6.19±0.2°, 7.45±0.2°, 16.23±0.2°, 18.28±0.2°, 18.95±0.2°, 26.31±0.2°, 27.04±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00032 No. 2θ (±0.2°) 1 4.889 2 6.188 3 7.452 4 9.717 5 9.968 6 12.225 7 14.529 8 15.022 9 15.814 10 16.226 11 18.28 12 18.954 13 19.822 14 20.923 15 21.821 16 22.575 17 23.384 18 26.307 19 27.035 20 28.12 21 29.98 22 35.366 23 38.437 further, the crystal form I of the compound of Formula (IV) has an XRPD pattern as shown in FIG. 25; further, the crystal form I of the compound of Formula (IV) has a differential scanning calorimetry (DSC) curve with an endothermic peak at 86.86° C.±3° C., and further has a DSC pattern as shown in FIG. 26; further, the crystal form I of the compound of Formula (IV) has a thermogravimetric analysis (TGA) curve with a weight loss of 1.298% at 46.81° C.±3° C., a weight loss of up to 3.607% at 89.20° C.±3° C. and a weight loss of up to 4.641% at 169.65° C.±3° C., and further has a TGA pattern as shown in FIG. 27.

16. The compound of Formula (IV), or the crystal form H, K, I or J thereof according to claim 12, the crystal form J of the compound of Formula (IV) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 4.97±0.2°, 6.19±0.2°, 16.33±0.2°, 19.15±0.2°, 19.84±0.2°, 21.02±0.2°, 22.68±0.2°, 23.92±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00033 No. 2θ (±0.2°) 1 4.967 2 6.191 3 9.94 4 11.773 5 14.569 6 15.043 7 15.739 8 16.326 9 18.378 10 19.148 11 19.839 12 21.021 13 21.728 14 22.679 15 23.448 16 23.922 17 26.328 18 27.037 19 28.043 20 28.682 21 29.895 22 30.996 23 34.409 24 39.37 further, the crystal form J of the compound of Formula (IV) has an XRPD pattern as shown in FIG. 28; further, the crystal form J of the compound of Formula (IV) has a differential scanning calorimetry (DSC) curve with an endothermic peak at 61.29° C.±3° C., an endothermic peak at 86.40° C.±3° C., and an endothermic peak at 151.50° C.±3° C., and further has a DSC pattern as shown in FIG. 29; further, the crystal form J of the compound of Formula (IV) has a thermogravimetric analysis (TGA) curve with a weight loss of 3.412% at 220.12° C.±3° C., and further has a TGA pattern as shown in FIG. 30.

17. The compound of Formula (IV), or the crystal form H, K, I or J thereof according to claim 12, wherein, for the crystal form H or K of the compound of Formula (IV), the compound of Formula (IV) is represented by Compound (IV-1): ##STR00028## wherein, for the crystal form I or J of the compound of Formula (IV), the compound of Formula (IV) is represented by Compound (IV-2): ##STR00029##

18. A compound of Formula (V) represented by the following formula, or a crystal form L thereof: ##STR00030## wherein, n.sub.5 is selected from 0.5 and 1; m.sub.5 is selected from 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5 and 4; wherein the crystal form L of the compound of Formula (V) has an X-ray powder diffraction (XRPD) pattern with characteristic diffraction peaks at 2θ angles of 10.39±0.2°, 18.04±0.2°, 20.31±0.2°.

19. The compound of Formula (V), or the crystal form L thereof according to claim 18, wherein the crystal form L of the compound of Formula (V) has an XRPD pattern with characteristic diffraction peaks at 2θ angles of 7.91±0.2°, 10.39±0.2°, 14.18±0.2°, 16.01±0.2°, 16.47±0.2°, 18.04±0.2°, 20.31±0.2°, 21.91±0.2°, and further has an XRPD pattern analysis data as shown in the following table: TABLE-US-00034 No. 2θ (±0.2°) 1 7.906 2 10.393 3 11.788 4 13.626 5 14.18 6 15.049 7 15.774 8 16.012 9 16.466 10 17.164 11 18.044 12 19.86 13 20.311 14 20.829 15 21.91 16 22.538 17 23.194 18 24.59 19 25.124 20 25.417 21 25.894 22 26.35 23 28.104 24 28.632 25 29.821 26 31.395 27 32.48 28 34.257 29 35.399 30 39.383 further, the crystal form L of the compound of Formula (V) has an XRPD pattern as shown in FIG. 34; further, the crystal form L of the compound of Formula (V) has a differential scanning calorimetry (DSC) curve with an endothermic peak at 168.08° C.±3° C., an endothermic peak at 204.17° C.±3° C., and further has a DSC pattern as shown in FIG. 35; further, the crystal form L of the compound of Formula (V) has a thermogravimetric analysis (TGA) curve with a weight loss of 0.830% at 80.19° C.±3° C., a weight loss of up to 3.058% at 149.87° C.±3° C., and a weight loss of up to 4.648% at 201.25° C.±3° C., and further has a TGA pattern as shown in FIG. 36; further, for the crystal form L of the compound of Formula (V), the compound of Formula (V) is represented by Compound V-1: ##STR00031##

20. A method of treating an influenza virus-associated disease, comprising administering a therapeutically effective amount of the compound or the crystal form thereof according to claims 1 to a subject in need thereof.

21. A method of treating an influenze virus-associated disease, comprising administering a therapeutically effective amount of the compound or the crystal form thereof according to claim 4 to a subject in need thereof.

22. A method of treating an influenza virus-associated disease, comprising administering a therapeutically effective amount of the compound or the crystal form thereof according to claim 7 to a subject in need thereof.

23. A method of treating an influenza virus-associated disease, comprising administering a therapeutically effective amount of the compound or the crystal form thereof according to claim 9 to a subject in need thereof.

24. A method of treating an influenza virus-associated disease, comprising administering a therapeutically effective amount of the compound or the crystal form thereof according to claim 12 to a subject in need thereof.

25. A method of treating an influenza virus-associated disease, comprising administering a therapeutically effective amount of the compound or the crystal form thereof according to claim 18 to a subject in need thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1: A Cu-Kα radiated XRPD pattern of the crystal form A;

(2) FIG. 2: A DSC pattern of the crystal form A;

(3) FIG. 3: A TGA pattern of the crystal form A;

(4) FIG. 4: A Cu-Kα radiated XRPD pattern of the crystal form B;

(5) FIG. 5: A DSC pattern of the crystal form B;

(6) FIG. 6: A TGA pattern of the crystal form B;

(7) FIG. 7: A Cu-Kα radiated XRPD pattern of the crystal form C;

(8) FIG. 8: A DSC pattern of the crystal form C;

(9) FIG. 9: A TGA pattern of the crystal form C;

(10) FIG. 10: A Cu-Kα radiated XRPD pattern of the crystal form D;

(11) FIG. 11: A DSC pattern of the crystal form D;

(12) FIG. 12: A TGA pattern of the crystal form D;

(13) FIG. 13: A Cu-Kα radiated XRPD pattern of the crystal form E;

(14) FIG. 14: A DSC pattern of the crystal form E;

(15) FIG. 15: A TGA pattern of the crystal form E;

(16) FIG. 16: A Cu-Kα radiated XRPD pattern of the crystal form F;

(17) FIG. 17: A DSC pattern of the crystal form F;

(18) FIG. 18: A TGA pattern of the crystal form F;

(19) FIG. 19: A Cu-Kα radiated XRPD pattern of the crystal form G;

(20) FIG. 20: A DSC pattern of the crystal form G;

(21) FIG. 21: A TGA pattern of the crystal form G;

(22) FIG. 22: A Cu-Kα radiated XRPD pattern of the crystal form H;

(23) FIG. 23: A DSC pattern of the crystal form H;

(24) FIG. 24: A TGA pattern of the crystal form H;

(25) FIG. 25: A Cu-Kα radiated XRPD pattern of the crystal form I;

(26) FIG. 26: A DSC pattern of the crystal form I;

(27) FIG. 27: A TGA pattern of the crystal form I;

(28) FIG. 28: A Cu-Kα radiated XRPD pattern of the crystal form J;

(29) FIG. 29: A DSC pattern of the crystal form J;

(30) FIG. 30: A TGA pattern of the crystal form J;

(31) FIG. 31: A Cu-Kα radiated XRPD pattern of the crystal form K;

(32) FIG. 32: A DSC pattern of the crystal form K;

(33) FIG. 33: A TGA pattern of the crystal form K;

(34) FIG. 34: A Cu-Kα radiated XRPD pattern of the crystal form L;

(35) FIG. 35: A DSC pattern of the crystal form L

(36) FIG. 36: A TGA pattern of the crystal form L.

DETAILED DESCRIPTION

(37) To better understand the present disclosure, hereinafter it is further described by reference to specific examples. However, the present disclosure is not limited to the specific embodiments.

Reference Example 1: Preparation of Compound BB-1

(38) ##STR00015##

(39) Step 1: Synthesis of Compound BB-1-2:

(40) To a solution of Compound BB-1-1 (300 mg, 1.97 mmol) in bromoform (5 mL) was added t-butyl nitrite (406 mg, 3.94 mmol). The mixture was stirred at 60° C. for 1 hr, and then stirred at 90° C. for 1 hr. The reaction mixture was cooled to room temperature, and concentrated to give a crude product, which was purified by flash chromatography with silica gel (5-20% ethyl acetate/petroleum ether) to give Compound BB-1-2 (300.00 mg, yield: 70.50%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 11.25 (br s, 1H), 8.54 (dd, J=1.88, 2.64 Hz, 1H), 7.69 (dd, J=2.51, 7.28 Hz, 1H). MS (ESI) m/z: 215.9 (M+H.sup.+).

(41) Step 2: Synthesis of Compound BB-1-3:

(42) To a solution of Compound BB-1-2 (300 mg, 1.39 mmol) in N,N-dimethylformamide (5 mL) was added triphenylmethyl chloride (426 mg, 1.53 mmol) and potassium carbonate (576 mg, 4.17 mmol). The mixture was stirred at 25° C. for 12 hrs. The reaction mixture was diluted with ethyl acetate (50 mL), and washed with saturated brine (15 mL×3). The organic phase was dried over anhydrous sodium sulfate, concentrated to give a crude product, which was purified by flash chromatograph with silica gel (0-10% ethyl acetate/petroleum ether) to give Compound BB-1-3 (350 mg, yield: 54.94%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 8.16 (dd, J=1.25, 2.76 Hz, 1H), 7.53 (dd, J=3.01, 7.53 Hz, 1H), 7.25 (s, 15H). MS (ESI) m/z: 458.2 (M+H.sup.+).

(43) Step 3: Synthesis of Compound BB-1:

(44) To a solution of Compound BB-1-3 (350 mg, 763.66 μmol) and Bis(pinacolato)diboron (291 mg, 1.15 mmol) in N,N-dimethylformamide (7 mL) was added potassium acetate (225 mg, 2.29 mmol) and r-bis(di-tert-butylphosphine) palladium ferrocene dichloride (28 mg, 38.18 μmol). The mixture was stirred at 100° C. under nitrogen protection for 2 hr. The reaction mixture was cooled to room temperature and then filtered. The filtrate was diluted with ethyl acetate (50 mL), and washed with saturated brine (20 mL×3). The organic phase was dried over anhydrous sodium sulfate, and concentrated to give a crude product, which was purified by flash chromatograph with silica gel (0-10% ethyl acetate/petroleum ether) to give BB-1 (300 mg, yield: 77.73%). MS (ESI) m/z: 733.2 (M+Na.sup.+).

Example 1: Preparation of Compound 1

(45) ##STR00016##

(46) Step 1: Synthesis of Compound 1-2

(47) At 0° C., Compound 1-1 (25.00 g, 149.73 mmol) was dissolved into glycol dimethyl ether (80 mL), and cyclopropylmagnesium bromide (0.5 M, 500.10 mL) was dropwise added. The reaction mixture was stirred at room temperature overnight. Then, the reaction mixture was cooled to 0° C., and a solution of triethylamine (15.15 g, 149.73 mmol, 20.75 mL) in tetrafuran (30 mL) and a solution of iodine (38.00 g, 149.73 mmol) in tetrafuran (30 mL) were respectively added. The reaction mixture was stirred at room temperature for 3 hr. To the reaction mixture was added ethyl acetate (1 L), washed with water (300 mL×3) and saturated brine (300 mL), respectively, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was purified by silica gel column (petroleum ether) to give Compound 1-2 (8 g, yield: 25.8%).

(48) Step 2: Synthesis of Compound 1-3

(49) Compound (2S,3S)-ethyl 3-aminobiscyclo[2.2.2]octane-2-carboxylate (450 mg, 2.28 mmol) and Compound 1-2 (450 mg, 2.17 mmol) were dissolved into tetrahydrofuran (5.00 mL), and diisopropylethylamine (841.35 mg, 6.51 mmol) was added. The reaction mixture was stirred at 55° C. for 3 hr. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by flash column with silica gel (petroleum ether: ethyl acetate=10:1 to 5:1) to give Compound 1-3 (460.00 mg, yield: 57.6%).

(50) Step 3: Synthesis of Compound 1-4

(51) At room temperature, Compounds 1-3 (460.00 mg, 1.25 mmol) and BB-1 (1.05 g, 1.25 mmol) were dissolved into 2-methyltetrahydrofuran (8.00 mL) and water (2.00 mL), and potassium phosphate (796.34 mg, 3.75 mmol), tri(dibenzalacetone)dipalladium (114.51 mg, 125.05 μmol) and 2-biscyclohexylphosphine-2′,4′,6′-triisopropylbiphenyl (119 mg, 250 μmol) were respectively added. The reaction mixture was reacted at 80° C. overnight. The reaction mixture was cooled to room temperature, and water (30 mL) was added. Then, the mixture was filtered, and the filtrate was extracted with ethyl acetate (10 mL×3). The organic phases were combined, and dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was purified by flash column with silica gel (petroleum ether: ethyl acetate=20:1 to 3:1) to give Compound 1-4 (600 mg, yield: 61%). MS (ESI) m/z: 773.4 (M+H.sup.+).

(52) Step 4: Synthesis of Compound 1-5

(53) At room temperature, Compound 1-4 (600.00 mg, 844.11 μmol) was dissolved into dichloromethane (6.00 mL), and trifluoroacetic acid (962.45 mg, 8.44 mmol) and triethyl hydrosilane (981.53 mg, 8.44 mmol) were added. The reaction mixture was reacted at room temperature for 4 hr. The reaction mixture was concentrated under reduced pressure, and the obtained crude product was purified by flash column with silica gel (petroleum ether: ethyl acetate=10:1 to 2:1) to give Compound 1-5 (350.00 mg, yield: 87.6%). MS (ESI) m/z: 469.2 (M+H.sup.+).

(54) Step 5: Synthesis of Compound 1

(55) At room temperature, Compound 1-5 (160.00 mg, 341.52 μmol) was dissolved into dioxane (3.00 mL) and water (500.00 μL), and sodium hydroxide (136.61 mg, 3.42 mmol) was added. The reaction mixture was reacted at 80° C. for 1 hr. The reaction mixture was concentrated under reduced pressure and then adjusted to pH=5 by adding 1 M HCl. The precipitated solid was filtered, and the filter cake was washed with water (10 mL), and dried to give 1 (55.4 mg, yield: 36.5%). .sup.1H NMR (400 MHz, d.sub.4-MeOH) δ 8.49-8.58 (m, 2H), 4.92 (br s, 1H), 2.78 (br d, J=6.78 Hz, 1H), 2.22-2.31 (m, 1H), 2.11 (br s, 1H), 1.80-2.02 (m, 4H), 1.61-1.77 (m, 3H), 1.44-1.59 (m, 2H), 1.25-1.34 (m, 3H), 1.03-1.11 (m, 2H). MS m/z: 441.1 [M+1].sup.+.

Example 2: Preparation of Crystal Form A

(56) 100 mg of Compound 1 was placed into a glass flask, and 0.8 mL of ethanol was added to form a suspension. The suspension sample was placed in a thermomixer (40° C.) for conducting an experiment (in dark). The suspension sample was shaken at 40° C. for 60 hr, and centrifuged. Then, the residual sample was dried in a vacuum drying oven (40° C.) overnight, to give the crystal form A. The obtained crystal form A has an XRPD pattern as shown in FIG. 1, a DSC pattern as shown in FIG. 2, and a TGA pattern as shown in FIG. 3.

Example 3: Preparation of Crystal Form A

(57) 100 mg of Compound 1 was placed into a glass flask, and 1.2 mL of ethyl acetate was added to form a suspension. The suspension sample was placed in a thermomixer (40° C.) for conducting an experiment (in dark). The suspension sample was shaken at 40° C. for 60 hr, and centrifuged. Then, the residual sample was dried in a vacuum drying oven (40° C.) overnight, to give the crystal form A which was substantially consistent with that of Example 2.

Example 4: Preparation of Crystal Form B

(58) 66 g of Compound 1 was added into a mixed solution (600 mL) of ethanol and water (ethanol: water=1:1) to form a suspension. The suspension was placed on a stirrer, stirred at 40° C. for 48 hr, and filtered. The filter cake was oven-dried to give the crystal form B. The obtained crystal form B has an XRPD pattern as shown in FIG. 4, a DSC pattern as shown in FIG. 5, and a TGA pattern as shown in FIG. 6.

Example 5: Preparation Crystal Form B

(59) 66 g of Compound 1 was added into a mixed solution (600 mL) of ethanol and water (ethanol: water=3:1) to form a suspension. The suspension was placed on a stirrer and stirred at 40° C. for 48 hr, and filtered. The filter cake was oven-dried to give the crystal form B which was substantially consistent with that of Example 4.

Example 6: Preparation of Crystal Form C

(60) 5 g of Compound 1 was added into a 250 mL eggplant-shaped flask, THF (100 mL) was added, and hydrochloric acid (0.98 mL, dissolved in 9 mL THF) was added. The mixture was stirred at 30° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give the crystal form C (4.29 g). The obtained crystal form C has an XRPD pattern as shown in FIG. 7, a DSC pattern as shown in FIG. 8, and a TGA pattern as shown in FIG. 9.

Example 7: Preparation of Crystal Form D

(61) Crystal form C (0.201 g) was dissolved into acetonitrile (3 mL) and beaten. The mixture was stirred at 30° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give the crystal form D. The obtained crystal form D has an XRPD pattern as shown in FIG. 10, a DSC pattern as shown in FIG. 11, and a TGA pattern as shown in FIG. 12.

Example 8: Preparation of Crystal Form E

(62) 5 g of Compound 1 was added into a 250 mL eggplant-shaped flask, THF (100 mL) was added, and p-toluenesulfonic acid monohydrate (2.26 g, dissolved in 10 mL THF) was added. The mixture was stirred at 30° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give a solid (0.425 g). The solid (0.101 g) was added into acetone (2 mL) and beaten for 12h to give the crystal form E. The obtained crystal form E has an XRPD pattern as shown in FIG. 13, a DSC pattern as shown in FIG. 14, and a TGA pattern as shown in FIG. 15.

Example 9: Preparation of Crystal Form F

(63) 5 g of Compound 1 was added into a 250 mL eggplant-shaped flask, THF (100 mL) was added, and an aqueous NaOH solution (0.477 g, dissolved in 1 mL water) was added. The mixture was stirred at 30° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give the crystal form F. The obtained crystal form F has an XRPD pattern as shown in FIG. 16, a DSC pattern as shown in FIG. 17, and a TGA pattern as shown in FIG. 18.

Example 9-1: Preparation of Crystal Form F

(64) 202 mg of the crystal form F obtained in Example 9 was added into EtOH:H.sub.2O=3:1 (4 mL). The mixture was stirred at 30° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give the crystal form F. The obtained crystal form F was substantially consistent with the crystal form F of Example 9.

Example 10: Preparation of Crystal Form G

(65) Crystal form F (0.206 g) was dissolved into acetonitrile (3 mL) and beaten. The mixture was stirred at 30° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give the crystal form G. The obtained crystal form G has an XRPD pattern as shown in FIG. 19, a DSC pattern as shown in FIG. 20, and a TGA pattern as shown in FIG. 21.

Example 11: Preparation of Crystal Form H

(66) About 2 g of Compound 1 was added into a 100 mL eggplant-shaped flask, THF (35 mL) was added, and an aqueous KOH solution (0.255 g, dissolved in 0.5 mL and 5 mL THF) was added. The mixture was stirred at 30° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give the crystal form H. The obtained crystal form H has an XRPD pattern as shown in FIG. 22, a DSC pattern as shown in FIG. 23, and a TGA pattern as shown in FIG. 24.

Example 12: Preparation of Crystal Form I

(67) Crystal form H (0.201 g) was dissolved into acetonitrile (3 mL) and beaten. The mixture was stirred at 25° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give the crystal form I. The obtained crystal form I has an XRPD pattern as shown in FIG. 25, a DSC pattern as shown in FIG. 26, and a TGA pattern as shown in FIG. 27.

Example 13: Preparation of Crystal Form J

(68) Crystal form H (0.202 g) was dissolved into acetone (3 mL) and beaten. The mixture was stirred at 25° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give the crystal form J. The obtained crystal form J has an XRPD pattern as shown in FIG. 28, a DSC pattern as shown in FIG. 29, and a TGA pattern as shown in FIG. 30.

Example 14: Preparation of Crystal Form K

(69) Crystal form H (0.201 g) was dissolved into a mixed solvent of ethanol and water (ethanol: water=3:1) (4 mL) and beaten. The mixture was stirred at 25° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give the crystal form K. The obtained crystal form K has an XRPD pattern as shown in FIG. 31, a DSC pattern as shown in FIG. 32, and a TGA pattern as shown in FIG. 33.

Example 15: Preparation of Crystal Form L

(70) 2 g of Compound 1 was added into a 100 mL eggplant-shaped flask, THF (35 mL) was added, and an aqueous solution of calcium hydroxide (0.168 g, dissolved in 0.5 mL and 5 mL THF) was added. The mixture was stirred at 25° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give a solid (1.440 g). The solid (0.204 g) was dissolved into a mixed solvent of ethanol and water (ethanol: water=3:1) (4 mL) and beaten. The mixture was stirred at 25° C. for 12 hr, and the solid was filtered. The filter cake was dried under vacuum at 40° C. to give the crystal form L. The obtained crystal form L has an XRPD pattern as shown in FIG. 34, a DSC pattern as shown in FIG. 35, and a TGA pattern as shown in FIG. 36.

Experimental Example 1: Solid Stability Test of Crystal Form A

(71) A sample of crystal form A was placed at the bottom of a glass bottle to form a thin layer. The sample was placed under high temperature, high humidity and acceleration conditions. The bottle was sealed with an aluminum foil, and some small holes were pierced on the aluminum foil to ensure that the sample could fully contact with the ambient air. The sample placed under light radiation was placed upright at room temperature and open to the air. The sample was exposed to a light source, and radiated with sufficient energy prior to taking samples for detection. Samples were taken at various time points for analysis, and the detection results were compared with the initial detection results obtained at Day 0. The investigation items include appearance, content and impurities. The test results are shown in the following table:

(72) TABLE-US-00015 Total Impurities Test Conditions Time Point Appearance Content (%) (%) XPRD — Day 0 Off-white solid 98.51 1.49 Crystal form A High temperature Day 5 Off-white solid Not detected Not detected Not detected (60° C., open) Day 10 Off-white solid 98.25 1.75 Crystal form A High Humidity Day 5 Off-white solid Not detected Not detected Not detected (room Day 10 Off-white solid 98.25 1.75 Crystal form A temperature/relative humidity 92.5%, open) Light Radiation Day 5 Off-white solid Not detected Not detected Not detected (total illumination: 1.2 × Day 10 Off-white solid 96.89 3.11 Crystal form A 10.sup.6 Lux .Math. hr/) Acceleration Test Day 5 Off-white solid Not detected Not detected Not detected (40° C./relative humidity Day 10 Off-white solid 98.36 1.64 Crystal form A 75%, open)

(73) It can be seen from the above test results that the crystal form A prepared in the aforesaid examples shows that there is a relatively small change in total impurity content under the conditions of high temperature, high humidity and accelerated experiments. The XRPD detection method found that the crystal form A does not change and has a relatively high stability.

Experimental Example 2: Solid Stability Test of Crystal Form B

(74) A sample of the crystal form B was placed at the bottom of a glass bottle to form a thin layer. The sample was placed under high temperature, high humidity and acceleration conditions. The bottle was sealed with an aluminum foil, and some small holes were pierced on the aluminum foil to ensure that the sample could fully contact with the ambient air. The sample placed under light radiation was placed upright at room temperature and open to the air. The sample was exposed to a light source, and radiated with sufficient energy prior to taking samples for detection. Samples were taken at various time points for analysis, and the detection results were compared with the initial detection results obtained at Day 0. The investigation items include appearance, content and impurities. The test results are shown in the following table:

(75) TABLE-US-00016 Total Test Conditions Time points Appearance Content (%) Impurities (%) XPRD — Day 0 Off-white 98.89 1.11 Crystal form B solid High temperature Day 5 Off-white 98.91 1.09 Not detected (60° C., open) solid Day 10 Off-white 98.89 1.11 Crystal form B solid High Humidity Day 5 Off-white 98.92 1.08 Not detected (room solid temperature/relative Day 10 Off-white 98.90 1.10 Crystal form B humidity 92.5%, open) solid Light Radiation Day 5 Off-white Not detected Not detected Not detected (total illumination: 1.2 × solid 10.sup.6 Lux .Math. hr/) Day 10 Off-white 98.03 1.97 Crystal form B solid

(76) It can be seen from the above test results that the crystal form B prepared in the aforesaid examples shows that there is almost no change in total impurity content under the conditions of high temperature, high humidity, and a relatively small change in total impurity in the accelerated experiments. The XRPD detection method found that the crystal form B does not change, and has a relatively high stability.

Experimental Example 3

(77) The stability of the crystal form D was tested by the same method as that of Experimental Example 1. Samples were taken at various time points, and the test results were compared with the initial test results obtained at Day 0. The investigation items include appearance, impurities, and crystal forms. The test results are shown in the following table:

(78) TABLE-US-00017 Total Test Conditions Time Points Appearance Impurities (%) XPRD — Day 0 Off-white solid 3.3 Crystal form D High temperature Day 5 Off-white solid Not detected Not detected (60° C., open) Day 12 Off-white solid 3.3 Crystal form D High Humidity Day 5 Off-white solid Not detected Not detected (room Day 12 Off-white solid 3.4 Crystal form D temperature/relative humidity 92.5%, open) Light Radiation Day 5 Off-white solid Not detected Not detected (total illumination: 1.2 × Day 12 Off-white solid 3.4 Crystal form D 10.sup.6 Lux .Math. hr/) Acceleration Test Day 5 Off-white solid Not detected Not detected (40° C./relative humidity Day 12 Off-white solid 3.5 Crystal form D 75%, open)

(79) It can be seen from the above test results that the crystal form D prepared in the aforesaid examples shows that there is almost no change in total impurity content under the conditions of high temperature, high humidity, light radiation and accelerated experiments. The XRPD detection method found that the crystal form D does not change, and has a relatively high stability.

Experimental Example 4

(80) The stability of the crystal form F was tested by the same method as that of Experimental Example 1. Samples were taken at various time points, and the test results were compared with the initial test results obtained at Day 0. The investigation items include appearance, impurities, and crystal forms. The test results are shown in the following table:

(81) TABLE-US-00018 Total Test Conditions Time Points Appearance Impurities (%) XPRD — Day 0 Off-white solid 1.7 Crystal form F High temperature Day 5 Off-white solid Not detected Not detected (60° C., open) Day 12 Off-white solid 1.9 Crystal form F High Humidity Day 5 Off-white solid Not detected Not detected (room Day 12 Off-white solid 1.9 Crystal form F temperature/relative humidity 92.5%, open) Light Radiation Day 5 Off-white solid Not detected Not detected (total illumination: 1.2 × Day 12 Off-white solid 1.8 Crystal form F 10.sup.6 Lux .Math. hr/) Acceleration Test Day 5 Off-white solid Not detected Not detected (40° C./relative humidity Day 12 Off-white solid 1.9 Crystal form F 75%, open)

(82) It can be seen from the above test results that the crystal form F prepared in the aforesaid examples shows that there is almost no change in total impurity content under the conditions of high temperature, high humidity and accelerated experiments. The XRPD detection method found that the crystal form F does not change, and has a relatively high stability.

Experimental Example 5

(83) The stability of the crystal form G was tested by the same method as that of Experimental Example 1. Samples were taken at various time points, and the test results were compared with the initial test results obtained at Day 0. The investigation items include appearance, impurities, and crystal forms. The test results are shown in the following table:

(84) TABLE-US-00019 Total Test Conditions Time Points Appearance Impurities (%) XPRD — Day 0 Off-white solid 0.35 Crystal form G High temperature Day 5 Off-white solid Not detected Not detected (60° C., open) Day 30 Off-white solid 0.42 Crystal form G High Humidity Day 5 Off-white solid Not detected Not detected (room temperature/relative Day 30 Off-white solid 0.33 Crystal form G humidity 92.5%, open) Light Radiation Day 5 Off-white solid Not detected Not detected (total illumination: 1.2 × Day 12 Yellowish solid 0.54 Crystal form G 10.sup.6 Lux .Math. hr/) Acceleration Test Day 5 Off-white solid Not detected Not detected (40° C./relative humidity Day 30 Off-white solid 0.35 Crystal form G 75%, open)

(85) It can be seen from the above test results that the crystal form G prepared in the aforesaid examples shows that there is almost no change in total impurity content under the conditions of high humidity and accelerated experiments, and there is a relatively small change in total impurity content under the conditions of high temperature. The XRPD detection method found that the crystal form G does not change, and thus it can be known that the crystal form has a relatively high stability.

Experimental Example 6

(86) The stability of the crystal form H was tested by the same method as that of Experimental Example 1. Samples were taken at various time points, and the test results were compared with the initial test results obtained at Day 0. The investigation items include appearance, impurities, and crystal forms. The test results are shown in the following table:

(87) TABLE-US-00020 Total Test Conditions Time Point Appearance Impurities (%) XPRD — Day 0 Off-white solid 1.8 Crystal form H High temperature Day 5 Off-white solid Not detected Not detected (60° C., open) Day 30 Off-white solid 2.0 Crystal form H High Humidity Day 5 Off-white solid Not detected Not detected (room Day 30 Off-white solid 1.9 Crystal form H temperature/relative humidity 92.5%, open) Light Radiation Day 5 Off-white solid Not detected Not detected (total illumination: 1.2 × Day 12 Yellowish solid 1.9 Crystal form H 10.sup.6 Lux .Math. hr/) Acceleration Test Day 5 Off-white solid Not detected Not detected (40° C./relative humidity Day 30 Off-white solid 2.0 Crystal form H 75%, open)

(88) It can be seen from the above test results that the crystal form H prepared in the aforesaid examples shows that there is almost no change in total impurity content under the conditions of high temperature, high humidity, light radiation and accelerated experiments. The XRPD detection method found that the crystal form H does not change, and it can be seen that the crystal form has a relatively high stability.

(89) Those skilled in the art can understand that the crystal forms in the examples are obtained by long-term stirring and beating/crystallizing, and tend to form a stable state, thereby having a relatively high stability. They have considerable pharmaceutical prospects, and can also be used as an intermediate in the preparation of pharmaceutical products in production.

(90) Biological Part

(91) Influenza Virus Cytopathy (CPE) Experiment

(92) The antiviral activity of a compound against influenza virus (IFV) is evaluated by measuring the half effective concentration (EC.sub.50) value of a compound. The cytopathic test is widely used to determine the protective effect of the compound on virus-infected cells to reflect the antiviral activity of the compound.

(93) Influenza Virus CPE Experiment

(94) MDCK cells (ATCC, Catalog No. CCL-34) were seeded into a black 384-well cell culture plate at a density of 2,000-3,000 cells/well, and then placed in a 37° C., 5% CO.sub.2 incubator overnight. The compounds were diluted by use of Echo555 Non-Contact nanoliter-grade sonic pipetting system, and added into the wells (3-fold dilution, 8 test concentration points). Influenza virus A/Weiss/43 (H1N1) strain (ATCC, Catalog No. VR-96) was then added at 1-2 90% tissue culture infectious dose per well (TCID90) into the wells to allow that the final concentration of DMSO in the medium was 0.5%. Virus control wells (DMSO and virus added, but no compound added) and cell control wells (DMSO added, and no compound and virus added) were set. The plate was placed in a 37° C., 5% CO.sub.2 incubator for 5 days. After culturing for 5 days, a cell viability detection kit CCK8 was used to detect the cell viability. The raw data was used to calculate the antiviral activity of the compound.

(95) The antiviral activity of the compound is represented by the inhibition rate (%) of the compound on the cytoviral effect caused by the virus. The calculation formula is as follows:

(96) $\%\mathrm{Inhibition}\mathrm{Rate}=\left(\frac{\mathrm{Sample}\mathrm{value}-\mathrm{Average}\mathrm{value}\mathrm{of}\mathrm{virus}\mathrm{controls}}{\begin{array}{c}\mathrm{Average}\mathrm{value}\mathrm{of}\mathrm{cell}\mathrm{controls}-\\ \mathrm{Average}\mathrm{value}\mathrm{of}\mathrm{virus}\mathrm{controls}\end{array}}\right)\times 100$

(97) GraphPad Prism software was used to perform a nonlinear fitting analysis on the inhibition rate of the compound to give the EC.sub.50 value of the compound. The experimental results are shown in Table 15.

(98) TABLE-US-00021 TABLE 15 In vitro screening test results Compound EC.sub.50 (nM) Compound 1 0.013

(99) Results and Discussion: Compound 1 shows a positive effect in the experiment of inhibiting influenza virus replication at a cell level.

Experimental Example 2: In Vivo Drug Efficacy Studies

(100) Evaluation of the efficacy of compounds in influenza A virus H1N1 mouse infection model

(101) Mice were infected with Influenza A virus H1N1 (Virapur Company, Catalog No.: F1003A) by intranasal drip, and were treated with the compound at 36 hr after infection. The mice were orally administered for 7 consecutive days, twice a day. By observing the changes in body weight and survival rate of mice, the anti-influenza A virus H1N1 effect of the compound in this model was evaluated.

(102) The experiment used SPF-grade BALB/c mice (Shanghai Lingchang Biological Technology Co., Ltd.), 6-7 weeks of age, female. The mice adapted to the BSL-2 animal room for at least 3 days and then the experiment started. The infection day was set as Day 0. The mice were anesthetized by intraperitoneal injection of pentobarbital sodium (75 mg/kg, 10 ml/kg). The animal was infected with the H1N1 A/WSN/33 virus by intranasal drip after it entered the state of deep anesthesia, and the infection volume was 50 μl. From Day 1 to Day 7, 10 mg/kg (administration volume of 10 ml/kg) of the test compound was administered orally twice a day. The time of the first administration was 36 hr after infection. The state of the mice was observed daily, and the weight and survival rate of mice were recorded. At Day 14, all the surviving animals were euthanized.

(103) The measured survival rate and weight loss rate of the animals are shown in Table 16.

(104) TABLE-US-00022 TABLE 16 Measured survival rate and rate of weight loss of animals Rate of Weight Loss Survival Rate Compound (Day 9) (%) Compound 1 4.8% 100%