Honokiol Crystal Forms, Amorphous Form And Preparation Thereof
20220204432 · 2022-06-30
Assignee
Inventors
Cpc classification
A61K9/19
HUMAN NECESSITIES
B82Y5/00
PERFORMING OPERATIONS; TRANSPORTING
A61K9/127
HUMAN NECESSITIES
C07C39/21
CHEMISTRY; METALLURGY
A61K47/26
HUMAN NECESSITIES
C07C39/21
CHEMISTRY; METALLURGY
International classification
A61K9/127
HUMAN NECESSITIES
Abstract
The present invention provides honokiol crystal forms A, B and C and amorphous form and a preparation method thereof, wherein the preparation method comprises dissolving honokiol in n-heptane and standing at room temperature overnight to obtain the separated crystal as honokiol crystal form A; heating the honokiol crystal form A and cooling to room temperature to obtain a solid as honokiol crystal form B; heating the honokiol crystal form A to the molten state and then standing at quenching temperature to obtain the separated crystal as honokiol crystal form C; and adding the honokiol crystal form A with an antisolvent in DMSO/H.sub.2O system, and thus obtaining the oily matter as honokiol amorphous form. The honokiol crystal forms A, B and C and the amorphous form of the present invention have the advantages of good solubility, good stability, low hygroscopicity, long-term storage and/or good reproducibility, and are suitable for drug development.
Claims
1. A honokiol crystal form A, wherein a X-ray powder diffraction pattern of the honokiol crystal form A comprises characteristic peaks at 2theta values of 6.79°±0.2°, 9.10°±0.2°, 13.97°±0.2°, 14.97°±0.2° and 17.54°±0.2° under Cu—Kα radiation.
2. The honokiol crystal form A of claim 1, wherein the X-ray powder diffraction pattern of the honokiol crystal form A further comprises characteristic peaks at 2theta values of 20.61°±0.2°, 22.08°±0.2°, 24.01°±0.2° under Cu—Kα radiation.
3. A preparation method for the honokiol crystal form A of claim 1, comprising any one method selected from method I to method VIII: method I: dissolving honokiol in an organic solvent at 78° C. to 85° C. to obtain a honokiol solution, rapidly pouring the honokiol solution into another container, standing at room temperature overnight, centrifuging to collect the solids to obtain the honokiol crystal form A, wherein the organic solvent is n-heptane, petroleum ether, cyclohexane, n-hexane, toluene, DCM, DMSO, NMP, chloroform, methanol/water, or ethanol/water; method II: placing honokiol in a container, then adding an organic solvent to another container, placing the opened container containing the honokiol in the container containing the organic solvent, sealing and standing at room temperature, centrifuging to collect the solids to obtain the honokiol crystal form A, wherein the organic solvent is ethyl acetate, isopropyl acetate, tert-butyl methyl ether, chloroform, N-methylpyrrolidone, ethanol or acetonitrile; method III: placing honokiol in a container, dissolving in a solvent and filtering to obtain the clear honokiol solution, adding an antisolvent to another container, placing the opened container containing the clear honokiol solution in the container containing the antisolvent, sealing and standing at room temperature, centrifuging to collect the solids to obtain the honokiol crystal form A, wherein the solvent is ethanol, tetrahydrofuran, chloroform, ethyl acetate or isopropanol, and the antisolvent is n-hexane or water; method IV: placing honokiol in a container, adding an organic solvent to obtain a suspension, magnetically stir the obtained suspension at room temperature, centrifuging to collect the solids to obtain the honokiol crystal form A, wherein the organic solvent is methanol/water, acetonitrile/water, acetone/water, or dichloromethane/n-hexane; method V: placing honokiol in a container, adding an organic solvent to obtain a suspension, magnetically stir the obtained suspension at 40-60° C., centrifuging to collect the solids to obtain the honokiol crystal form A, wherein the organic solvent is n-heptane, Tween/n-heptane, chloroform/n-heptane, or isopropanol/water; method VI: placing honokiol in a container, adding an organic solvent to obtain a suspension, magnetically stir the obtained suspension at a temperature cycling, centrifuging to collect the solids to obtain the honokiol crystal form A, Where the temperature cycling includes 2 cycles of 60° C..fwdarw.5° C., 0.1° C./min, and 5° C..fwdarw.60° C., 1.5° C./min, wherein the organic solvent is n-heptane, ethanol/water, acetone/water, tetrahydrofuran/water, acetonitrile/water, ethyl acetate/n-heptane, tert-butyl methyl ether/n-hexane, or methyl isobutyl ketone/n-hexane; method VII: placing honokiol in a container, dissolving in an organic solvent, filtering and taking the filtrate, sealing the container containing the filtrate with a sealing membrane, making small holes on the sealing membrane, standing at room temperature to slowly volatilize, centrifuging to collect the solids to obtain the honokiol crystal form A, wherein the organic solvent is chloroform, dichloromethane, methanol, isopropanol, or methyl isobutyl ketone; method VIII: placing honokiol in a container, dissolving in a solvent, adding an antisolvent under magnetic stirring while adding dropwise, standing, centrifuging to collect the solids to obtain the honokiol crystal form A, wherein the solvent is ethanol, acetone, methyl isobutyl ketone, ethyl acetate, tert-butyl methyl ether, acetonitrile, dichloromethane or chloroform, and the antisolvent is n-heptane or water.
4. A honokiol crystal form B, wherein a X-ray powder diffraction pattern of the honokiol crystal form B comprises characteristic peaks at 2theta values of 6.15°±0.2°, 6.76°±0.2°, 8.96°±0.2° and 15.90°±0.2° under Cu—Kα radiation.
5. The honokiol crystal form B of claim 4, wherein the X-ray powder diffraction pattern of the honokiol crystal form B further comprises characteristic peaks at 2theta values of 17.49°±0.2° and 18.55°±0.2° under Cu—Kα radiation.
6. A preparation method for the honokiol crystal form B of claim 4, comprising the following steps of: heating a honokiol crystal form A to 80-82° C. and then cooling to the room temperature, and thus obtaining the solid as the honokiol crystal form B.
7. A honokiol crystal form C, wherein a X-ray powder diffraction pattern of the honokiol crystal form C comprises characteristic peaks at 2theta values of 14.04°±0.2°, 15.84°±0.2°, 17.42°±0.2° and 19.48°±0.2° under Cu—Kα radiation.
8. The honokiol crystal form C of claim 7, wherein the X-ray powder diffraction pattern of the honokiol crystal form C further comprises characteristic peaks at 2theta values of 21.26°±0.2°, 23.10°±0.2° and 24.06°±0.2° under Cu—Kα radiation.
9. A preparation method for the honokiol crystal form C of claim 7, comprising the following steps of: (1) heating honokiol crystal form A to a molten state at 83-100° C. and stirring; (2) placing the melt obtained in Step (1) rapidly at a quenching temperature of −20 to −196° C. to obtain a crystal, and separating the crystal as the honokiol crystal form C.
10. The preparation method of claim 9, wherein the quenching temperature of Step (2) is −80 to −196° C.
11. An honokiol amorphous form, wherein the honokiol amorphous form has an X-ray powder diffraction pattern as shown in
12. A preparation method for honokiol amorphous form of claim 11, comprising the following steps of: (1) dissolving honokiol crystal form A of in DMSO to obtain a solution; (2) adding antisolvent water to the solution obtained in Step (1) and stirring; (3) centrifuging, discarding the supernatant, standing at room temperature, and thus obtaining an oily matter as the honokiol amorphous form.
13. A drug composition or drug product, comprising an effective amount of the honokiol crystal form A of claim 1, and pharmaceutical excipients.
14. A honokiol nano-liposome lyophilized powder, comprising an effective amounts of the honokiol crystal form A of claim 1.
15. A preparation method for the honokiol nano-liposome lyophilized powder of claim 14, comprising the following steps of: dissolving the honokiol crystal form A, phospholipid, phosphatidylethanolamine and cholesterol in absolute ethanol to obtain a solution, injecting the solution that is completely dissolved into the purified water, stirring for rotary evaporation to remove the ethanol, adding a lyophilized excipient, and freeze-drying to obtain the honokiol nano-liposome lyophilized powder.
16. (canceled)
17. (canceled)
18. A drug composition or drug product, comprising an effective amount of the honokiol crystal form B of claim 4 and pharmaceutical excipients.
19. A drug composition or drug product, comprising an effective amount of the honokiol crystal form C of claim 7 and pharmaceutical excipients.
20. A drug composition or drug product, comprising an effective amount of the honokiol amorphous form of claim 11, and pharmaceutical excipients.
21. A honokiol nano-liposome lyophilized powder, comprising an effective amount of the honokiol crystal form B of claim 4.
22. A honokiol nano-liposome lyophilized powder, comprising an effective amount of the honokiol crystal form C of claim 7.
23. A honokiol nano-liposome lyophilized powder, comprising an effective amount of the honokiol amorphous form of claim 11.
24. A preparation method for the honokiol nano-liposome lyophilized powder of claim 21, comprising the following steps of: dissolving the honokiol crystal form B, phospholipid, phosphatidylethanolamine and cholesterol in absolute ethanol to obtain a solution, injecting the solution that is completely dissolved into the purified water, stirring for rotary evaporation to remove the ethanol, adding a lyophilized excipient, and freeze-drying to obtain the honokiol nano-liposome lyophilized powder.
25. A preparation method for the honokiol nano-liposome lyophilized powder of claim 22, comprising the following steps of: dissolving the honokiol crystal form C, phospholipid, phosphatidylethanolamine and cholesterol in absolute ethanol to obtain a solution, injecting the solution that is completely dissolved into the purified water, stirring for rotary evaporation to remove the ethanol, adding a lyophilized excipient, and freeze-drying to obtain the honokiol nano-liposome lyophilized powder.
26. A preparation method for the honokiol nano-liposome lyophilized powder of claim 23, comprising the following steps of: dissolving the honokiol amorphous form, phospholipid, phosphatidylethanolamine and cholesterol in absolute ethanol to obtain a solution, injecting the solution that is completely dissolved into the purified water, stirring for rotary evaporation to remove the ethanol, adding a lyophilized excipient, and freeze-drying to obtain the honokiol nano-liposome lyophilized powder.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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DETAILED DESCRIPTION
[0094] The following embodiments further illustrate the present invention, however, they do not constitute a restrictions or limitation of the scope of the invention.
[0095] Reagents used in embodiments of the present invention are conventional reagents in the field. Unless otherwise specified hereinafter, the reagents used in the present invention are well known in the field, but they are described in the present invention in detail as much as possible.
[0096] Raw materials used in the embodiment and the preparation method of honokiol powder are detailed in Journal of Chromatography A, 1142 (2007) 115-122.
Embodiment 1: Preparing the Honokiol Crystal Form a by Recrystallization with n-Heptane
[0097] Dissolving 201.25 mg honokiol powder completely in n-heptane at 80° C., then pouring the solution to another beaker quickly, standing at the room temperature overnight, and thus obtaining the crystals as the honokiol crystal form A, of which the XRPD pattern is shown in
[0098] The corresponding values of 2theta values and intensities of the honokiol crystal form A in the embodiment are shown in Table 1:
TABLE-US-00001 TABLE 1 Corresponding Values of 2theta Values and Intensities of the Honokiol Crystal Form A No. 2theta Intensity % 1 6.0935 4.95 2 6.5328 16.41 3 6.7971 100 4 9.1031 9.68 5 12.1559 2.25 6 13.9719 9.78 7 14.9720 4.12 8 16.5075 1.41 9 17.5405 21.51 10 19.3106 1.6 11 19.5697 2.11 12 20.6195 8.33 13 20.9485 4.39 14 21.4800 5.3 15 22.0843 12.89 16 22.8333 4.1 17 22.9755 3.79 18 23.7021 3.91 19 24.0117 7.96 20 24.5432 4.73 21 25.0433 4.66 22 25.5378 3.06 23 25.8289 2.58 24 26.0495 3.15 25 27.2453 1.76 26 27.6398 4.35 27 28.2230 4.18 28 29.0779 1.63 29 29.3534 3.65 30 30.0155 2.14 31 30.3107 2 32 30.8450 1.41 33 31.5797 1.11 34 31.8736 1.12 35 33.7018 1.18 36 34.6063 1.37 37 35.3406 2.01 38 36.2867 0.91 39 37.4284 0.98 40 37.8896 0.88 41 38.7965 0.59 42 40.5974 0.49 43 42.0185 0.79 44 45.2149 0.3 45 49.5154 0.29 46 51.6076 0.21
Embodiment 2: Preparing the Honokiol Crystal Form a by Gas-Solid Permeation
[0099] Weighing about 15 mg of honokiol powder into a 3 mL vial, adding about 3 mL of N-methylpyrrolidone into another 20 mL vial, placing the 3 mL opened vial in the 20 mL vial and sealing the 20 mL vial. Standing at room temperature until the surface of honokiol is slightly wet, centrifuging to collect the solids, and thus obtaining the honokiol crystal form A. The obtained solid is the honokiol crystal form A as confirmed by an XRPD test, of which the XRPD pattern is shown in
Embodiment 3: Preparing the Honokiol Crystal Form a by Gas-Liquid Permeation
[0100] Weighing about 15 mg of honokiol powder into a 3 mL vial, dissolving in 0.5 mL of isopropanol and filtering through a 0.45 μm PTFE membrane to obtain a clear solution; adding about 3 mL of antisolvent water to another 20 mL vial, placing the 3 mL opened vial containing the clear solution in the 20 mL vial, sealing the 20 mL vial and standing at the room temperature for 9 days. Centrifuging to collect the solids, and thus obtaining the honokiol crystal form A. The obtained solid is the honokiol crystal form A as confirmed by an XRPD test, of which the XRPD pattern is shown in
Embodiment 4: Preparing the Honokiol Crystal Form a by Suspension Stirring at Room Temperature
[0101] Weighing about 15 mg honokiol powder into a 3 mL vial, adding 0.3-0.5 mL of acetonitrile/water (1:9), magnetically stirring the obtained suspension at the room temperature for 4 days, centrifuging to collect the solids, and thus obtaining the honokiol crystal form A. The obtained solid is the honokiol crystal form A as confirmed by an XRPD test, of which the XRPD pattern is shown in
Embodiment 5: Preparing the Honokiol Crystal Form a by Suspension Stirring at 50° C.
[0102] Weighing about 15 mg of honokiol powder into a 3 mL vial, adding 0.5 mL of chloroform/n-heptane (1:9), magnetically stirring the obtained suspension below 50° C. and at 1000 rpm for 4 days, centrifuging to collect the solids, and thus obtaining the honokiol crystal form A. The obtained solid is the honokiol crystal form A as confirmed by an XRPD test, of which the XRPD pattern is shown in
Embodiment 6: Preparing the Honokiol Crystal Form a by Temperature Cycling Stirring
[0103] Weighing about 15 mg of honokiol powder into a 3 mL vial, adding 0.5 mL of ethanol/water (1:9), magnetically stirring the obtained suspension at the temperature cycling (2 cycles including 50° C..fwdarw.5° C., 0.1° C./min, and 5° C..fwdarw.50° C., 1.5° C./min) and at 1000 rpm for 16 h, centrifuging to collect the solids, and thus obtaining the honokiol crystal form A. The obtained solid is the honokiol crystal form A as confirmed by an XRPD test, of which the XRPD pattern is shown in
Embodiment 7: Preparing the Honokiol Crystal Form a by Slow Volatilization
[0104] Weighing about 15 mg of honokiol powder into a 3 mL vial, dissolving in 1.0 mL of chloroform, filtering (with PTFE membrane with a pore size of 0.45 μm) and taking the filtrate. Sealing the vial containing clear solution with a sealing membrane, making several small holes on the sealing membrane, standing at room temperature to slowly volatilize for separation of solids, centrifuging to collect the solids, and thus obtaining the honokiol crystal form A. The obtained solid is the honokiol crystal form A as confirmed by an XRPD test, of which the XRPD pattern is shown in
Embodiment 8: Preparing the Honokiol Crystal Form a by Antisolvent Addition
[0105] Weighing about 15 mg of honokiol powder into a 3 mL vial, dissolving in 0.5-1.0 mL of ethanol, adding n-heptane under magnetic stirring, stirring while adding dropwise, standing overnight, centrifuging to collect the solids, and thus obtaining the honokiol crystal form A. The obtained solid is the honokiol crystal form A as confirmed by an XRPD test, of which the XRPD pattern is shown in
Embodiment 9: Preparation of the Honokiol Crystal Form B
[0106] Placing the sample at the room temperature immediately after heating 200.75 mg of honokiol crystal form A to 80° C., and thus obtaining the solid as the honokiol crystal form B, of which the XRPD spectrum is shown in
[0107] The corresponding values of 2theta values and intensities of the honokiol crystal form B in the embodiment are shown in Table 2:
TABLE-US-00002 TABLE 2 Corresponding Values of 2theta Values and Intensities of the Honokiol Crystal Form B No. 2theta Intensity % 1 6.1504 100.00 2 6.7608 15.08 3 8.9653 4.93 4 13.0808 1.88 5 13.9526 2.77 6 14.3170 1.39 7 15.9024 6.34 8 17.2149 3.10 9 17.4955 7.62 10 17.9023 2.02 11 18.5502 6.26 12 19.5855 2.79 13 21.2438 2.88 14 22.7821 4.06 15 23.2199 2.99 16 23.5529 1.90 17 24.1781 2.76 18 24.5335 1.66 19 24.8318 1.84 20 26.7765 0.88 21 28.1861 1.33 22 29.3438 0.75 23 30.3149 0.58 24 31.3429 0.99 25 31.9348 0.60 26 37.6766 0.24
Embodiment 10: Preparation of the Honokiol Crystal Form B
[0108] Placing the sample at the room temperature immediately after heating 200.65 mg of honokiol crystal form A to 80° C. and holding for 1 h, and thus obtaining the solid as the honokiol crystal form B, of which the XRPD spectrum is shown in
[0109] The corresponding values of 2theta values and intensities of the honokiol crystal form B in the embodiment are shown in Table 3:
TABLE-US-00003 TABLE 3 Corresponding Values of 2theta Values and Intensities of the Honokiol Crystal Form B No. 2theta Intensity % 1 6.1145 100 2 8.9191 4.83 3 13.0492 1.62 4 13.9199 2.4 5 14.2635 1.4 6 15.8646 6.33 7 17.1657 2.82 8 17.4554 6.9 9 17.8548 2.25 10 18.5209 7.66 11 19.5406 2.55 12 21.1944 2.98 13 22.733 4.21 14 23.1809 2.91 15 24.1315 2.66 16 24.7949 2.72 17 25.5231 1.15 18 28.0686 1.32 19 31.3283 1.03 20 37.6203 0.39
Embodiment 11: Preparation of the Honokiol Crystal Form B
[0110] Placing the sample at the room temperature immediately after heating 200.25 mg of honokiol crystal form A to 80° C. and holding for 4 h, and thus obtaining the solid as the honokiol crystal form B, of which the XRPD spectrum is shown in
[0111] The corresponding values of 2theta values and intensities of the honokiol crystal form B in the embodiment are shown in Table 4:
TABLE-US-00004 TABLE 4 Corresponding Values of 2theta Values and Intensities of the Honokiol Crystal Form B No. 2theta Intensity % 1 6.1917 100 2 6.906 1.62 3 9.0082 2.3 4 12.3598 0.69 5 13.1118 0.79 6 13.9898 1.53 7 14.3439 0.65 8 15.9382 2.9 9 17.242 1.41 10 17.5357 3.32 11 17.9295 1.17 12 18.5849 6.77 13 19.6136 1.71 14 21.2758 1.66 15 22.796 2.94 16 23.3027 1.77 17 23.5706 0.92 18 24.211 1.3 19 24.5713 0.89 20 24.8569 2.3 21 25.7046 0.69 22 26.8102 0.51 23 27.5878 0.83 24 27.846 0.82 25 28.1965 1.3 26 29.2644 0.48 27 29.4753 0.48 28 30.4566 0.43 29 31.4005 0.71 30 31.9042 0.74 31 32.4824 0.32 32 33.0914 0.35 33 34.4144 0.23 34 37.6543 0.67 35 40.0415 0.16 36 41.6417 0.12 37 44.2058 0.11 38 44.6725 0.24 39 45.4141 0.1 40 46.396 0.08 41 49.6005 0.06 42 50.9781 0.15
Embodiment 12: Preparation of the Honokiol Crystal Form B
[0112] Placing the sample at the room temperature immediately after heating 201.25 mg of honokiol crystal form A to 80° C. and holding for 8 h, and thus obtaining the solid as the honokiol crystal form B, of which the XRPD spectrum is shown in
[0113] The corresponding values of 2theta values and intensities of the honokiol crystal form B in the embodiment are shown in Table 5:
TABLE-US-00005 TABLE 5 Corresponding Values of 2theta Values and Intensities of the Honokiol Crystal Form B No. 2theta Intensity % 1 6.0859 89.22 2 6.2586 100 3 8.9298 4.87 4 12.3175 1.75 5 13.0913 2.39 6 13.9301 3.52 7 14.4509 1.84 8 15.8914 7.98 9 17.1827 3.65 10 17.4718 8.24 11 17.8736 3.23 12 18.4936 8.26 13 18.6688 5.88 14 19.5528 4.02 15 21.2134 4.17 16 22.7336 6.36 17 23.309 5.07 18 23.551 3.42 19 24.1664 4.01 20 24.491 2.68 21 24.7726 3.54 22 25.6034 2.24 23 26.7734 1.89 24 27.5516 2.85 25 28.1247 3.39 26 29.4852 1.76 27 30.349 1.66 28 31.3386 2.25 29 31.9246 1.77 30 33.0757 1.12 31 34.4329 0.88 32 37.5659 1.08 33 39.9031 0.49 34 41.5763 0.43 35 44.6686 0.66 36 45.4135 0.37 37 46.3251 0.3 38 47.6436 0.24 39 49.5829 0.18 40 50.9185 0.28
Embodiment 13: Preparation of the Honokiol Crystal Form B
[0114] Placing the sample at the room temperature immediately after heating 200.15 mg of honokiol crystal form A to 82° C., and thus obtaining the solid as the honokiol crystal form B, of which the XRPD spectrum is shown in
[0115] The corresponding values of 2theta values and intensities of the honokiol crystal form B in the embodiment are shown in Table 6:
TABLE-US-00006 TABLE 6 Corresponding Values of 2theta Values and Intensities of the Honokiol Crystal Form B No. 2theta Intensity % 1 6.1607 59.56 2 6.5174 4.42 3 6.8724 100 4 9.0088 2.83 5 9.1692 2.01 6 12.2552 0.15 7 13.0794 0.21 8 14.006 1.83 9 14.2782 0.18 10 15.0641 1.05 11 15.9119 1.2 12 16.5522 0.02 13 17.2202 0.8 14 17.4603 3.35 15 17.6241 3.47 16 17.9209 0.63 17 18.1087 0.95 18 18.5873 2.86 19 19.4273 0.48 20 19.6235 0.56 21 20.5742 2.29 22 20.7317 3.11 23 21.0086 0.44 24 21.2764 0.5 25 21.4974 0.46 26 22.0504 1.83 27 22.7693 1.11 28 23.2588 0.34 29 23.9398 0.89 30 24.1083 1.51 31 24.5834 0.41 32 24.8612 0.65 33 25.1094 0.98 34 25.5801 0.25 35 25.9375 0.42 36 27.6025 1.62 37 27.7528 1.62 38 28.0915 0.81 39 28.3164 0.99 40 29.3251 0.53 41 29.9711 0.25 42 30.4174 0.5 43 30.8365 0.03 44 31.3583 0.17 45 31.6421 0.29 46 31.9099 0.2 47 32.3041 0.06 48 33.1748 0.13 49 33.7905 0.13 50 34.4223 0.11 51 35.4539 0.53 52 36.381 0.12 53 37.4516 0.32 54 38.7171 0.05 55 39.5517 0.21 56 40.4904 0.03 57 42.0234 0.46 58 43.1904 0.04 59 44.0785 0.06 60 44.6456 0.13 61 45.3062 0.06 62 46.4142 0.09 63 47.6515 0.02 64 49.4697 0.18 65 51.7075 0.16 66 53.7499 0.04 67 57.3282 0.02
Embodiment 14: Preparation of the Honokiol Crystal Form C
[0116] Heating 1002.5 mg of honokiol crystal form A to the molten state at 85° C. in a crucible, then quickly placing the crucible in a refrigerator to maintain −80° C., standing overnight, and thus obtaining the solid as the honokiol crystal form C, of which the XRPD spectrum is shown in
[0117] The corresponding values of 2theta values and intensities of the honokiol crystal form C in the embodiment are shown in Table 7:
TABLE-US-00007 TABLE 7 Corresponding Values of 2theta Values and Intensities of the Honokiol Crystal Form C No. 2theta Intensity % 1 9.40 6.34 2 13.18 10.95 3 14.20 20.17 4 15.94 40.35 5 17.52 100 6 19.70 30.55 7 21.26 45.82 8 22.34 46.97 9 23.14 67.44 10 24.10 66.57 11 25.82 36.02 12 27.62 33.14 13 29.34 26.22 14 30.16 25.36 15 31.56 27.67 16 34.50 22.48
Embodiment 15: Preparation of the Honokiol Crystal Form C
[0118] Heating 998.3 mg of honokiol crystal form A to the molten state at 85° C. in a crucible, then quickly placing the crucible in liquid nitrogen (−196° C.), standing overnight, and thus obtaining the solid as the honokiol crystal form C, of which the XRPD spectrum is shown in
[0119] The corresponding values of 2theta values and intensities of the honokiol crystal form C in the embodiment are shown in Table 8:
TABLE-US-00008 TABLE 8 Corresponding Values of 2theta Values and Intensities of the Honokiol Crystal Form C No. 2theta Intensity % 1 9.20 7.76 2 13.14 10.56 3 14.18 22.98 4 15.88 39.13 5 17.42 100 6 19.48 32.76 7 21.28 46.12 8 22.18 47.20 9 23.10 74.07 10 24.10 57.76 11 25.70 36.80 12 27.62 30.90 13 29.28 26.86 14 30.18 23.60 15 31.44 24.53 16 34.54 18.17
Embodiment 16: Preparation of Honokiol Amorphous Form
[0120] Weighing about 15 mg of honokiol crystal form A into a 20 mL vial, dissolving in 0.5-1.0 mL of solvent DMSO, adding the antisolvent H.sub.2O under magnetic stirring, and stirring while adding dropwise. Centrifuging, discarding the supernatant, standing at room temperature, and thus obtaining an oily matter as the amorphous, of which the XRPD pattern is shown in
Embodiment 17: TGA/DSC Tests for Honokiol Crystal Forms a, B and C
[0121] Thermogravimetric analysis (TGA): Instrument Manufacturer: METTLER-TOLEDO; Instrument Model: TGA/DSC2/1600; Test Conditions: taking an appropriate amount of trial-production sample under nitrogen atmosphere at 30-350° C., raising the temperature at a constant speed of 10° C./min, measuring changes in weight of the substance with the temperature, and plotting the weight-temperature change curve—TGA curve;
[0122] differential Scanning calorimeter (DSC): Instrument Manufacturer: METTLER-TOLEDO; Instrument Model: DSC3+/500; Test Conditions: taking an appropriate amount of trial-production sample under nitrogen atmosphere at 30-150° C., raising the temperature at a constant speed of 10° C./min, and determining the DSC curve of the sample;
[0123] heating the honokiol crystal forms A, B and C from room temperature to 150° C. respectively.
[0124] TGA results of the honokiol crystal form A show a 1.0% weight loss, and DSC results show two endothermic signals at 78.1° C. and 86.1° C. (peak temperature) as shown in
[0125] TGA results of the honokiol crystal form B show a 1.3% weight loss, and DSC results show an endothermic signal at 86.1° C. (peak temperature) as shown in
[0126] TGA results of the honokiol crystal form C show a 93.3% weight loss, and DSC results show an endothermic signal at 86.3° C. (peak temperature) as shown in
Embodiment 18: Solubility Tests for Honokiol Crystal Forms a, B and C
[0127] Since the solubility of the drug directly affects the dissolution and oral bioavailability of the drug preparation, the inventor investigated the solubility of the honokiol crystal forms A, B and C in PBS. Preparing respective saturated solutions of the honokiol crystal forms A, B and C, shaking at 37° C. for 4 h, and centrifuging and transferring the supernatant into a high performance liquid chromatograph. At the same time, preparing a standard solution of the honokiol (source: China Institutes for Food and Drug Control, Batch No.: 110730-201915, ID: EH2H-BMTW), and using the external standard method for quantification. Thus measured solubility of the honokiol crystal form A is 41.35 μg/ml, the solubility of the honokiol crystal form B is 43.66 μg/ml, and the solubility of the honokiol crystal form C is 48.12 μg/ml.
Embodiment 19: Photostability Tests for Honokiol Crystal Forms a, B and C
[0128] Standing the honokiol crystal forms A, B and C respectively in a small beaker in sunlight, and investigating the stability of each of the honokiol crystal forms A, B and C. Photographs of the honokiol crystal forms A, B and C after exposure to sunlight are shown in
Embodiment 20: Stress Tests for Honokiol Crystal Forms a, B and C
[0129] Treating the honokiol crystal forms A, B and C under high temperature (60° C.), high humidity (relative humidity 92.5%, 25° C.) and high-light irradiation (illuminance 45001x±5001x), respectively, diluting with methanol, and investigating the changes of related substances through testing with HPLC.
[0130] The purity of the honokiol crystal form A is 99.83% before the stress test, 99.63% after the high temperature test, 99.67% after the high humidity test, and 99.64% after intense light irradiation test respectively. The HPLCs of the honokiol crystal form A before the stress test is shown in
[0131] The purity of the honokiol crystal form B is 99.88% before the stress test, 99.70% after the high temperature test, 99.70% after the high humidity test, and 99.60% after the high-light irradiation test respectively. The HPLCs of the honokiol crystal form B before the stress test is shown in
[0132] The purity of the honokiol crystal form C is 99.86% before the stress test, 99.36% after the high temperature test, 99.59% after the high humidity test, and 99.31% after intense light irradiation test respectively. The HPLCs of the honokiol crystal form C before the stress test is shown in
Embodiment 21: Preparation of Honokiol Nano-Liposome Lyophilized Powder
[0133] All of the honokiol crystal forms A, B and C and the amorphous form are used to prepare the honokiol nano-liposome lyophilized powder with the same preparation method. Taking the honokiol crystal form B as an example, the preparation method is given below:
[0134] Dissolving 50 mg of honokiol crystal form B, 500 mg of soybean phospholipid, 200 mg of cholesterol and 200 mg of Cultured phosphatidylethanolamine in 50 mL of absolute ethanol, dissolving completely, injecting the solution into 300 mL of purified water, stirring for rotary evaporation to remove the ethanol, adding 800 mg of sucrose as the lyophilized excipient, and thus obtaining the honokiol nano-liposome lyophilized powder by freeze-drying. After reconstitution of the lyophilized powder, the particle size of honokiol nano-liposome is 114 nm as determined by a laser particle analyzer.