Magnolol crystal form, amorphous form and preparation method

Abstract

The invention provides honokiol crystal form A, crystal form B, crystal form C and amorphous form and a preparation method thereof. The method comprises: dissolving honokiol in n-heptane, standing at room temperature overnight, and the precipitated crystal is the honokiol crystal form A; the honokiol crystal form A is cooled to room temperature after heating, and the obtained solid is the honokiol crystal form B; the honokiol crystal form A is heated to a molten state, and then placed at a quenching temperature, and the precipitated crystal is the honokiol crystal form C; the oily substance obtained by adding the honokiol crystal form A with a DMSO / H2O system anti-solvent is the honokiol amorphous form. The honokiol crystal form A, crystal form B and crystal form C of the invention have good solubility, good stability, low hygroscopicity, can be stored for a long time, have good repeatability, and are suitable for drug development; the honokiol amorphous form of the invention has good solubility and is suitable for the development of pharmaceutical preparations.

Description

Technical Field

[0001] The invention relates to the field of medicine, and in particular to a crystal form and an amorphous form of honokiol and a preparation method thereof. Background Art

[0002] Honokiol, whose English name is Honokiol, has a chemical name of 3',5-di-2-propenyl-1,1'-biphenyl-2,4'-diphenol, and its structural formula is shown in the following formula (I):

[0003]

[0004] Honokiol is a small molecule compound extracted and separated from the bark of Magnolia officinalis Rehd.et Wils., which has a wide range of biological activities. Its main biological activities include antimicrobial, antioxidant, anti-anxiety, anti-depression and anti-thrombotic. In recent years, more and more studies have shown that honokiol has good anti-tumor activity, and its anti-tumor effect has the characteristics of multi-target, multi-effect and low toxicity and side effects. Animal experiments have shown that honokiol has an inhibitory effect on a variety of rapidly growing tumors and has a wide anti-tumor spectrum. Its mechanism of action is to induce tumor cell apoptosis, inhibit tumor cell migration and proliferation, and inhibit tumor angiogenesis, thereby exerting an anti-tumor effect.

[0005] The study of drug crystal forms is one of the important contents in new drug research. Therefore, it is of great significance to research and develop a more excellent phase of honokiol in terms of physicochemical properties and bioavailability. Summary of the invention

[0006] The object of the present invention is to provide a honokiol crystal form which is more excellent in solubility, stability, hygroscopicity and the like.

[0007] In the first aspect of the present invention, a honokiol crystal form is provided, hereinafter referred to as honokiol crystal form A. Using Cu-Kα radiation, the X-ray powder diffraction pattern of the honokiol crystal form A of the present invention has 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°.

[0008] Furthermore, using Cu-Kα radiation, the X-ray powder diffraction pattern of the honokiol crystal form A of the present invention has characteristic peaks at 2theta values ​​of 6.79°±0.2°, 9.10°±0.2°, 13.97°±0.2°, 14.97°±0.2°, 17.54°±0.2°, 20.61°±0.2°, 22.08°±0.2°, and 24.01°±0.2°.

[0009] Without limitation, the honokiol crystal form A of the present invention has Figure 1 , Figure 3-Figure 9 X-ray powder diffraction (XRPD) pattern shown.

[0010] Without limitation, the honokiol crystal form A of the present invention has Figure 2 Thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) spectra are shown.

[0011] The present invention also provides a method for preparing honokiol crystal form A, comprising:

[0012] Method 1: Preparation of Honokiol Crystal Form A by Recrystallization

[0013] The honokiol solution is obtained by dissolving honokiol in an organic solvent at 78°C to 85°C, and the honokiol solution is quickly poured into another container, left to stand overnight at room temperature, and the solid is collected by centrifugation to obtain:

[0014] The organic solvent is n-heptane, petroleum ether, cyclohexane, n-hexane, toluene, DCM, DMSO, NMP, chloroform, methanol / water, or ethanol / water;

[0015] Method 2: Preparation of Honokiol Crystal Form A by Gas-Solid Permeation Method

[0016] Put honokiol in a container, add an organic solvent to another container, place the container containing honokiol open in the container containing the organic solvent, seal, and let stand at room temperature (until the surface of honokiol is slightly moist), and collect the solid by centrifugation to obtain,

[0017] The organic solvent is ethyl acetate, isopropyl acetate, methyl tert-butyl ether, chloroform, N-methylpyrrolidone, ethanol or acetonitrile;

[0018] Method 3: Preparation of Honokiol Crystal Form A by Gas-Liquid Permeation Method

[0019] Put honokiol in a container, dissolve it with a positive solvent and filter it to obtain a clear honokiol solution. Take another container and add an anti-solvent into it. Put the container with the clear honokiol solution in the container containing the anti-solvent with its mouth open, seal it, let it stand at room temperature, and collect the solid by centrifugation to obtain,

[0020] The positive solvent may be ethanol, tetrahydrofuran, chloroform, ethyl acetate or isopropanol, and the anti-solvent may be n-hexane or water;

[0021] Method 4: Preparation of Honokiol Crystal Form A by Room Temperature Suspension Stirring Method

[0022] Place honokiol in a container, add an organic solvent, place the obtained suspension under room temperature with magnetic stirring, and collect the solid by centrifugation to obtain:

[0023] The organic solvent may be methanol / water (94:6, 84:16, 69:31, 42:58), acetonitrile / water (1:9), acetone / water (1:9), or dichloromethane / n-hexane (1:9);

[0024] Method 5: Preparation of Honokiol Crystal Form A by 40-60℃ Suspension Stirring Method

[0025] Place honokiol in a container, add an organic solvent, place the resulting suspension at 40-60°C with magnetic stirring, and collect the solid by centrifugation to obtain:

[0026] The organic solvent may be n-heptane, Tween / n-heptane (1:9), chloroform / n-heptane (1:9), or isopropanol / water (1:9);

[0027] Method 6: Preparation of Honokiol Crystal Form A by Temperature Cycling Stirring Method

[0028] Honokiol is placed in a container, an organic solvent is added, the obtained suspension is magnetically stirred under temperature cycle, and the solid is collected by centrifugation to obtain:

[0029] The temperature cycle is 60°C → 5°C, 0.1°C / min, 5°C → 60°C, 1.5°C / min, 2 cycles, and the preferred temperature cycle is 50°C → 5°C, 0.1°C / min, 5°C → 50°C, 1.5°C / min, 2 cycles.

[0030] The organic solvent may be n-heptane, ethanol / water (1:9), acetone / water (1:9), tetrahydrofuran / water (1:9), acetonitrile / water (1:9), ethyl acetate / n-heptane (1:9), methyl tert-butyl ether / n-hexane (1:9), or methyl isobutyl ketone / n-hexane (1:9);

[0031] Method 7: Preparation of Honokiol Crystal Form A by Slow Volatilization Method

[0032] Place honokiol in a container, add an organic solvent to dissolve it, filter and take the filtrate, seal the container with the filtrate with a sealing film, poke several small holes in the sealing film, let it stand at room temperature to evaporate slowly, and collect the solid by centrifugation to obtain,

[0033] The organic solvent may be chloroform, dichloromethane, methanol, isopropanol, or methyl isobutyl ketone;

[0034] Method 8: Preparation of Honokiol Crystal Form A by Anti-Solvent Addition Method

[0035] Place honokiol in a container, dissolve it with a positive solvent, add an anti-solvent under magnetic stirring, add dropwise while stirring, let stand, and collect the solid by centrifugation to obtain,

[0036] The positive solvent may be ethanol, acetone, methyl isobutyl ketone, ethyl acetate, methyl tert-butyl ether, acetonitrile, dichloromethane or chloroform, and the anti-solvent may be n-heptane or water.

[0037] In the second aspect of the present invention, a honokiol crystal form is provided, hereinafter referred to as honokiol crystal form B. Using Cu-Kα radiation, the X-ray powder diffraction pattern of the honokiol crystal form B of the present invention has characteristic peaks at 2theta values ​​of 6.15°±0.2°, 6.76°±0.2°, 8.96°±0.2°, and 15.90°±0.2°.

[0038] Further, using Cu-Kα radiation, the X-ray powder diffraction pattern of the honokiol crystal form B of the present invention has characteristic peaks at 2theta values ​​of 6.15°±0.2°, 6.76°±0.2°, 8.96°±0.2°, 15.90°±0.2°, 17.49°±0.2°, and 18.55°±0.2°.

[0039] In a non-limiting manner, the honokiol crystal form B of the present invention has Fig.10 , Figures 12 to 15 X-ray powder diffraction (XRPD) pattern shown.

[0040] In a non-limiting manner, the honokiol crystal form B of the present invention has Fig.11 Thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) spectra are shown.

[0041] The present invention also provides a method for preparing honokiol crystal form B, comprising the following steps:

[0042] The honokiol crystal form A is heated to 80-82°C and then cooled to room temperature, and the obtained solid is the honokiol crystal form B.

[0043] In the third aspect of the present invention, a honokiol crystal form is provided, hereinafter referred to as honokiol crystal form C. Using Cu-Kα radiation, the X-ray powder diffraction pattern of the honokiol crystal form C of the present invention has characteristic peaks at 2theta values ​​of 14.04°±0.2°, 15.84°±0.2°, 17.42°±0.2°, and 19.48°±0.2°.

[0044] Further, using Cu-Kα radiation, the X-ray powder diffraction pattern of the honokiol crystal form C of the present invention has characteristic peaks at 2theta values ​​of 14.04°±0.2°, 15.84°±0.2°, 17.42°±0.2°, 19.48°±0.2°, 21.26°±0.2°, 23.10°±0.2°, and 24.06°±0.2°.

[0045] In a non-limiting manner, the honokiol crystal form C of the present invention has Fig.16 and Fig.18 X-ray powder diffraction (XRPD) pattern shown.

[0046] In a non-limiting manner, the honokiol crystal form C of the present invention has Fig.17 Thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) spectra are shown.

[0047] The present invention also provides a method for preparing honokiol crystal form C, comprising the following steps:

[0048] (1) heating honokiol crystal form A at a temperature of 83-100° C. to a molten state, and stirring to mix thoroughly until no obvious particles are present;

[0049] (2) The melt obtained in step (1) is rapidly cooled to a temperature of -20 to -196°C, and the precipitated crystals are honokiol crystal form C.

[0050] Preferably, in the method for preparing honokiol crystal form C, the quenching temperature of step (2) is -80 to -196°C.

[0051] In the fourth aspect of the present invention, amorphous honokiol is provided. The amorphous honokiol of the present invention has good solubility and is suitable for the development of pharmaceutical preparations.

[0052] Without limitation, the amorphous form of honokiol of the present invention has Fig.19 X-ray powder diffraction (XRPD) pattern shown.

[0053] The present invention also provides a method for preparing amorphous honokiol, comprising the following steps:

[0054] (1) dissolving honokiol crystal form A in DMSO;

[0055] (2) adding anti-solvent water to the solution obtained in step (1) and stirring;

[0056] (3) Centrifuge, discard the supernatant, and let stand at room temperature to obtain an oily substance, which is amorphous honokiol.

[0057] Another object of the present invention is to provide a pharmaceutical composition or pharmaceutical preparation, which comprises a therapeutically effective amount of honokiol crystal form A, honokiol crystal form B, honokiol crystal form C or honokiol amorphous and pharmaceutical excipients. The pharmaceutical composition or pharmaceutical preparation is prepared in a manner well known in the pharmaceutical field, generally by mixing or contacting a therapeutically effective amount of honokiol crystal form A, honokiol crystal form B, honokiol crystal form C or honokiol amorphous with one or more pharmaceutical excipients to prepare the pharmaceutical composition or pharmaceutical preparation.

[0058] Another object of the present invention is to provide a kind of honokiol nanoliposome freeze-dried powder, which contains a therapeutically effective amount of honokiol crystal form A, honokiol crystal form B, honokiol crystal form C or honokiol amorphous. The honokiol nanoliposome freeze-dried powder can be prepared by the following method: dissolving honokiol crystal form A, honokiol crystal form B, honokiol crystal form C or honokiol amorphous (1 part), phospholipids (0.1-100 parts), phosphatidylethanolamine (0.01-100 parts) and cholesterol (0-100 parts) in anhydrous ethanol, after complete dissolution, injecting into purified water, stirring, rotary evaporation to remove ethanol, adding freeze-drying excipients (1-50 parts), freeze drying to prepare honokiol nanoliposome freeze-dried powder. After the freeze-dried powder is redissolved, the particle size of the honokiol nanoliposome is measured by a laser particle size analyzer to be 80-170nm.

[0059] The phospholipids are selected from one or more of soybean lecithin, egg yolk lecithin, hydrogenated soybean lecithin, hydrogenated egg yolk lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and its sodium salt, phosphatidylinositol, cardiolipin, sphingomyelin, and phosphatidylserine; the phosphatidylethanolamine is selected from peihua phosphatidylethanolamine, soybean phosphatidylethanolamine, distearoyl phosphatidylethanolamine-polyethylene glycol 600-20000 , dipalmitoylphosphatidylethanolamine-polyethylene glycol 600-20000 , dioleoylphosphatidylethanolamine-polyethylene glycol 600-20000 , Stearoyloleylphosphatidylethanolamine-polyethylene glycol 600-20000 , Stearoyllinoleylphosphatidylethanolamine-polyethylene glycol 600-20000 , Palmitoyloleoylphosphatidylethanolamine-polyethylene glycol 600-20000 , Palmitoyllinoleoylphosphatidylethanolamine-polyethylene glycol 600-20000 , Dimyristoylphosphatidylethanolamine-Polyethylene glycol 600-20000 , dilauroyl phosphatidylethanolamine-polyethylene glycol 600-20000 , didecanoylphosphatidylethanolamine-polyethylene glycol 600-20000 , dioctanoylphosphatidylethanolamine-polyethylene glycol 600-20000 or dihexanoylphosphatidylethanolamine-polyethylene glycol 600-20000One or more of; the excipient is selected from one or more of glucose, glycine, mannitol, inositol, sorbitol, sucrose, trehalose, lactose, galactose, glutamic acid, proline, lysine, and alanine.

[0060] Another object of the present invention is to provide the use of honokiol crystal form A, honokiol crystal form B, honokiol crystal form C or honokiol amorphous in the preparation of drugs for treating tumors.

[0061] Another object of the present invention is to provide the use of honokiol crystal form A, honokiol crystal form B, honokiol crystal form C or honokiol amorphous in the preparation of antimicrobial, antioxidant, antianxiety, antidepressant or antithrombotic drugs.

[0062] The honokiol crystal forms A, B and C of the present invention have good solubility, good stability, low hygroscopicity, can be stored for a long time, have good repeatability, and are suitable for drug development; the honokiol amorphous form of the present invention has good solubility and is suitable for the development of pharmaceutical preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0063] Figure 1 This is the XRPD spectrum of magnolia officinalis crystalline form A in Example 1.

[0064] Figure 2 This is the TGA / DSC spectrum of honokiol crystal form A in Example 1.

[0065] Figure 3 This is the XRPD spectrum of magnolia officinalis crystalline form A in Example 2.

[0066] Figure 4 This is the XRPD spectrum of magnolia officinalis crystalline form A in Example 3.

[0067] Figure 5 This is the XRPD spectrum of magnolia officinalis crystalline form A in Example 4.

[0068] Figure 6 This is the XRPD spectrum of magnolia officinalis crystalline form A in Example 5.

[0069] Figure 7 This is the XRPD spectrum of magnolia officinalis crystalline form A in Example 6.

[0070] Figure 8 This is the XRPD spectrum of magnolia officinalis crystalline form A in Example 7.

[0071] Fig. 9 This is the XRPD spectrum of magnolia officinalis crystalline form A in Example 8.

[0072] Fig.10 This is the XRPD spectrum of honokiol crystal form B after heating to 80°C and then immediately cooling to room temperature in Example 9.

[0073] Fig.11 This is the TGA / DSC spectrum of honokiol crystal form B after heating to 80°C and then immediately cooling to room temperature in Example 9.

[0074] Fig.12 This is the XRPD spectrum of honokiol crystal form B after heating to 80°C for 1 hour and then cooling to room temperature in Example 10.

[0075] Fig.13 This is the XRPD spectrum of honokiol crystal form B after heating to 80°C for 4 hours and then cooling to room temperature in Example 11.

[0076] Fig.14 This is the XRPD spectrum of honokiol crystal form B after heating to 80°C for 8 hours and then cooling to room temperature in Example 12.

[0077] Fig.15 This is the XRPD spectrum of honokiol crystal form B after heating to 82°C and then immediately cooling to room temperature in Example 13.

[0078] Fig.16 This is the XRPD spectrum of honokiol crystal form C after treatment at -80°C in Example 14.

[0079] Fig.17 This is the TGA / DSC spectrum of honokiol crystal form C after treatment at -80°C in Example 14.

[0080] Fig.18 This is the XRPD spectrum of honokiol crystal form C after treatment at -196°C in Example 15.

[0081] Fig.19 This is the XRPD pattern of the amorphous form of magnolol prepared in Example 16.

[0082] Fig. 20 These are photos of honokiol crystal form A, crystal form B and crystal form C after being exposed to sunlight.

[0083] Fig.21 This is the HPLC chart of honokiol crystal form A before the influencing factor experiment.

[0084] Fig. 22 This is the HPLC chart of honokiol crystal form A after high temperature test.

[0085] Fig.23 This is the HPLC chart of honokiol crystal form A after high humidity test.

[0086] Fig.24 This is the HPLC chart of honokiol crystal form A after strong light irradiation test.

[0087] Fig.25 This is the HPLC chart of honokiol crystal form B before the influencing factor experiment.

[0088] Fig.26 This is the HPLC chart of honokiol crystal form B after high temperature test.

[0089] Fig. 27 This is the HPLC chart of honokiol crystal form B after high humidity test.

[0090] Fig.28 This is the HPLC chart of honokiol crystal form B after strong light irradiation test.

[0091] Fig.29 This is the HPLC chart of honokiol crystal form C before the influencing factor experiment.

[0092] Fig.30 This is the HPLC chart of honokiol crystal form C after high temperature test.

[0093] Fig.31 This is the HPLC chart of honokiol crystal form C after high humidity test.

[0094] Fig.32 This is the HPLC chart of honokiol crystal form C after strong light irradiation test. Specific implementation plan

[0095] The following examples further illustrate the present invention; however, they are not intended to limit or define the scope of the present invention.

[0096] The reagents used in the embodiments of the present invention are conventional reagents in the art. Hereinafter, if not specifically stated, the reagents used in the present invention are well known in the art, but the present invention is still described in as much detail as possible.

[0097] The preparation method of the raw material and magnolol powder used in this example is detailed in Journal of Chromatography A, 1142 (2007) 115-122.

[0098] Example 1: Preparation of Honokiol Crystal Form A by Recrystallization Using n-Heptane

[0099] After 201.25 mg of honokiol powder was completely dissolved in 80°C n-heptane, it was quickly poured into another beaker and allowed to stand overnight at room temperature. The resulting crystals were honokiol crystal form A. Its XRPD spectrum is shown in Figure 1 , TGA / DSC spectrum see Figure 2 .

[0100] The corresponding values ​​of 2theta value and intensity of honokiol crystal form A in this embodiment are shown in Table 1:

[0101] Table 1 Corresponding values ​​of 2theta value and intensity of honokiol crystal form A

[0102]

[0103]

[0104] Example 2: Preparation of Honokiol Crystal Form A by Gas-Solid Permeation Method

[0105] Weigh about 15 mg of honokiol powder into a 3 mL vial, add about 3 mL of N-methylpyrrolidone to a 20 mL vial, place the 3 mL vial open in the 20 mL vial, and then seal the 20 mL vial. Let stand at room temperature until the sample surface is slightly moist, then centrifuge to collect the solid. XRPD test shows that the solid is honokiol crystal form A, and its XRPD spectrum is as follows: Figure 3 As shown, the TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form A prepared in Example 1.

[0106] Example 3: Preparation of Honokiol Crystal Form A by Gas-Liquid Permeation Method

[0107] Weigh about 15 mg of honokiol powder into a 3 mL vial, dissolve it with 0.5 mL of isopropanol, and filter it through a 0.45 μm PTFE filter membrane to obtain a clear solution; take another 20 mL vial and add about 3 mL of anti-solvent water to it, place the 3 mL vial containing the clear solution in the 20 mL vial, seal the 20 mL vial and let it stand at room temperature for 9 days. Collect the solid by centrifugation to obtain. After XRPD testing, the obtained solid is honokiol crystal form A, and its XRPD spectrum is as follows Figure 4 As shown, the TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form A prepared in Example 1.

[0108] Example 4: Preparation of Honokiol Crystal Form A by Room Temperature Suspension Stirring Method

[0109] Weigh about 15 mg of honokiol powder into a 3 mL vial, add 0.3-0.5 mL of acetonitrile / water (1:9), place the resulting suspension under magnetic stirring at room temperature for 4 days, and collect the solid by centrifugation. XRPD testing shows that the resulting solid is honokiol crystal form A, and its XRPD spectrum is as follows Figure 5 As shown, the TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form A prepared in Example 1.

[0110] Example 5: Preparation of Honokiol Crystal Form A by 50°C Suspension Stirring Method

[0111] About 15 mg of honokiol powder was weighed into a 3 mL vial, 0.5 mL of chloroform / n-heptane (1:9) was added, the resulting suspension was placed at 50°C and magnetically stirred at 1000 rpm for 4 days, and the solid was collected by centrifugation. XRPD test showed that the obtained solid was honokiol crystal form A, and its XRPD spectrum is as follows: Figure 6 As shown, the TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form A prepared in Example 1.

[0112] Example 6: Preparation of Honokiol Crystal Form A by Temperature Cycling Stirring Method

[0113] About 15 mg of honokiol powder was weighed into a 3 mL vial, and 0.5 mL of ethanol / water (1:9) was added. The resulting suspension was magnetically stirred at 1000 rpm for 16 h under temperature cycling (50°C → 5°C, 0.1°C / min, 5°C → 50°C, 1.5°C / min, 2 cycles), and the solid was collected by centrifugation. The obtained solid was honokiol crystal form A after XRPD testing, and its XRPD spectrum is shown as follows: Figure 7 As shown, the TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form A prepared in Example 1.

[0114] Example 7: Preparation of Honokiol Crystal Form A by Slow Evaporation Method

[0115] Weigh about 15 mg of honokiol powder into a 3 mL vial, add 1.0 mL of chloroform to dissolve, filter (PTFE filter membrane with a pore size of 0.45 μm) and take the filtrate. Seal the vial containing the clear solution with a sealing film, and poke several small holes in the sealing film. Let it stand at room temperature to slowly evaporate, precipitate solids, and collect the solids by centrifugation. After XRPD testing, the obtained solid is honokiol crystal form A, and its XRPD spectrum is as follows Figure 8 As shown, the TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form A prepared in Example 1.

[0116] Example 8: Preparation of Honokiol Crystal Form A by Anti-Solvent Addition Method

[0117] Weigh about 15 mg of honokiol powder into a 3 mL vial, dissolve it in 0.5-1.0 mL of ethanol, add n-heptane under magnetic stirring, add dropwise while stirring, let stand overnight, and collect the solid by centrifugation. XRPD test shows that the obtained solid is honokiol crystal form A, and its XRPD spectrum is as follows Fig. 9 As shown, the TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form A prepared in Example 1.

[0118] Example 9: Preparation of Honokiol Crystal Form B

[0119] After heating 200.75 mg of honokiol crystal form A to 80°C, the sample was immediately placed at room temperature. The resulting solid was honokiol crystal form B. Its XRPD spectrum is shown in Fig.10 , TGA / DSC spectrum see Fig.11 .

[0120] The corresponding values ​​of 2theta value and intensity of honokiol crystal form B in this embodiment are shown in Table 2:

[0121] Table 2 Corresponding values ​​of 2theta value and intensity of honokiol crystal form B

[0122]

[0123]

[0124] Example 10: Preparation of Honokiol Crystal Form B

[0125] After heating 200.65 mg of honokiol crystal form A to 80°C for 1 hour, the sample was immediately placed at room temperature. The resulting solid was honokiol crystal form B. Its XRPD spectrum is shown in Fig.12 The TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form B prepared in Example 9.

[0126] The corresponding values ​​of 2theta value and intensity of honokiol crystal form B in this embodiment are shown in Table 3:

[0127] Table 3 Corresponding values ​​of 2theta value and intensity of honokiol crystal form B

[0128] serial number 2theta strength% 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

[0129] Example 11: Preparation of Honokiol Crystal Form B

[0130] After heating 200.25 mg of honokiol crystal form A to 80°C for 4 hours, the sample was immediately placed at room temperature. The resulting solid was honokiol crystal form B. Its XRPD spectrum is shown in Fig.13 The TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form B prepared in Example 9.

[0131] The corresponding values ​​of 2theta value and intensity of honokiol crystal form B in this embodiment are shown in Table 4:

[0132] Table 4 Corresponding values ​​of 2theta value and intensity of honokiol crystal form B

[0133]

[0134]

[0135] Example 12: Preparation of Honokiol Crystal Form B

[0136] After heating 201.25 mg of honokiol crystal form A to 80°C for 8 hours, the sample was immediately placed at room temperature. The resulting solid was honokiol crystal form B. Its XRPD spectrum is shown in Fig.14 The TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form B prepared in Example 9.

[0137] The corresponding values ​​of 2theta value and intensity of honokiol crystal form B in this embodiment are shown in Table 5:

[0138] Table 5 Corresponding values ​​of 2theta value and intensity of honokiol crystal form B

[0139]

[0140]

[0141] Example 13: Preparation of Honokiol Crystal Form B

[0142] After heating 200.15 mg of honokiol crystal form A to 82°C, the sample was immediately placed at room temperature. The resulting solid was honokiol crystal form B. Its XRPD spectrum is shown in Fig.15 The TGA / DSC spectrum is consistent with the TGA / DSC spectrum of honokiol crystal form B prepared in Example 9.

[0143] The corresponding values ​​of 2theta value and intensity of honokiol crystal form B in this embodiment are shown in Table 6:

[0144] Table 6 Corresponding values ​​of 2theta value and intensity of honokiol crystal form B

[0145]

[0146]

[0147]

[0148] Example 14: Preparation of Honokiol Crystal Form C

[0149] 1002.5 mg of honokiol crystal form A was heated in a crucible at 85°C until it was molten, and then the crucible was quickly placed in a -80°C refrigerator and allowed to stand overnight. The resulting solid was honokiol crystal form C, and its XRPD spectrum is shown in Fig.16 , TGA / DSC spectrum see Fig.17 .

[0150] The corresponding values ​​of 2theta value and intensity of honokiol crystal form C in this embodiment are shown in Table 7:

[0151] Table 7 Corresponding values ​​of 2theta value and intensity of honokiol crystal form C

[0152]

[0153]

[0154] Example 15: Preparation of Honokiol Crystal Form C

[0155] 998.3 mg of honokiol crystal form A was heated in a crucible at 85°C until it was molten, and then the crucible was quickly placed in liquid nitrogen (-196°C) and allowed to stand overnight. The resulting solid was honokiol crystal form C, and its XRPD pattern is shown in Fig.18 The TGA / DSC spectrum is consistent with the TGA / DSC spectrum of magnolia bark phenol crystal form C prepared in Example 14.

[0156] The corresponding values ​​of 2theta value and intensity of honokiol crystal form C in this embodiment are shown in Table 8:

[0157] Table 8 Corresponding values ​​of 2theta value and intensity of honokiol crystal form C

[0158] serial number 2theta strength% 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

[0159] Example 16: Preparation of amorphous honokiol

[0160] Weigh about 15 mg of honokiol crystal form A into a 20 mL vial, dissolve it with 0.5-1.0 mL of positive solvent DMSO, add anti-solvent H2O under magnetic stirring, and stir while adding. Centrifuge, discard the supernatant, and stand at room temperature to obtain an amorphous oil. Its XRPD spectrum is shown in Fig.19 .

[0161] Example 17: TGA / DSC test of honokiol crystal form A, crystal form B and crystal form C

[0162] Thermogravimetric analysis (TGA): instrument manufacturer, METTLER-TOLEDO; instrument model, TGA / DSC2 / 1600; test conditions, take an appropriate amount of trial samples, use nitrogen atmosphere, increase the temperature at a constant rate of 10°C / min in the range of 30-350°C, measure the change of the weight of the substance with temperature, and draw a curve of weight-temperature change - TGA curve;

[0163] Differential Scanning Calorimeter (DSC): Instrument manufacturer, METTLER-TOLEDO; Instrument model, DSC3+ / 500; Test conditions, take an appropriate amount of trial sample, in nitrogen atmosphere, raise the temperature at a constant rate of 10°C / min in the range of 30-150°C, and measure the DSC curve of the sample;

[0164] Take honokiol crystal form A, crystal form B and crystal form C, and heat them from room temperature to 150° C. respectively.

[0165] The TGA results of honokiol crystal form A showed a weight loss of 1.0%, and the DSC results showed that it had two endothermic signals at 78.1°C and 86.1°C (peak temperature), see Figure 2 .

[0166] The TGA results of honokiol crystal form B showed a weight loss of 1.3%, and the DSC results showed that it had an endothermic signal at 86.1°C (peak temperature). Fig.11 .

[0167] The TGA results of honokiol crystal form C showed a weight loss of 93.3%; the DSC results showed that it had an endothermic signal at 86.3°C (peak temperature), Fig.17 .

[0168] Example 18: Solubility test of honokiol crystal form A, crystal form B and crystal form C

[0169] The solubility of the drug directly affects the dissolution and oral bioavailability of the drug preparation. The inventors investigated the solubility of honokiol crystal form A, crystal form B and crystal form C in PBS. Take honokiol crystal form A, crystal form B and crystal form C, each of which is prepared into a saturated solution, shaken at 37°C for 4h, and the supernatant is taken into a high-performance liquid chromatograph after centrifugation. At the same time, a reference solution of honokiol is prepared (the source of the reference substance is: China Food and Drug Inspection Institute, batch number: 110730-201915, ID: EH2H-BMTW), and the external standard method is used for quantification. The solubility of honokiol crystal form A is measured to be 41.35μg / ml, the solubility of honokiol crystal form B is 43.66μg / ml, and the solubility of honokiol crystal form C is 48.12μg / ml.

[0170] Example 19: Photostability test of honokiol crystal form A, crystal form B and crystal form C

[0171] Take honokiol crystal form A, crystal form B and crystal form C in small beakers and place them in the sun to investigate the stability of the three crystal forms in the sun. Fig. 20 .

[0172] Example 20: Test on factors affecting honokiol crystal form A, crystal form B and crystal form C

[0173] Take honokiol crystal form A, crystal form B and crystal form C, treat them under high temperature (60℃), high humidity (relative humidity 92.5%, 25℃), and strong light irradiation (illuminance of 4500lx±500lx), dilute them with methanol, inject them into HPLC for detection, and examine the changes of related substances.

[0174] Before the influencing factor experiment, the purity of honokiol crystal form A was 99.83%, after the high temperature test, the purity of honokiol crystal form A was 99.63%, after the high humidity test, the purity of honokiol crystal form A was 99.67%, and after the strong light irradiation test, the purity of honokiol crystal form A was 99.64%. The HPLC chart of honokiol crystal form A before the influencing factor experiment is shown in Fig.21 , the HPLC chart after high temperature test is shown in Fig. 22 , HPLC chart after high humidity test is shown in Fig.23 , the HPLC chart after strong light irradiation test is shown in Fig.24 .

[0175] Before the influencing factor experiment, the purity of honokiol crystal form B was 99.88%, after the high temperature test, the purity of honokiol crystal form B was 99.70%, after the high humidity test, the purity of honokiol crystal form B was 99.70%, and after the strong light irradiation test, the purity of honokiol crystal form B was 99.60%. The HPLC chart of honokiol crystal form B before the influencing factor experiment is shown in Fig.25 , the HPLC chart after high temperature test is shown in Fig.26 , HPLC chart after high humidity test is shown in Fig. 27 , the HPLC chart after strong light irradiation test is shown in Fig.28 .

[0176] The purity of honokiol crystal form C was 99.86% before the influencing factor experiment, 99.36% after the high temperature test, 99.59% after the high humidity test, and 99.31% after the strong light irradiation test. The HPLC chart of honokiol crystal form C before the influencing factor experiment is shown in Fig.29 , the HPLC chart after high temperature test is shown in Fig.30 , HPLC chart after high humidity test is shown in Fig.31 , the HPLC chart after strong light irradiation test is shown in Fig.32 .

[0177] Example 21: Preparation of honokiol nanoliposome freeze-dried powder

[0178] Honokiol crystal form A, crystal form B, crystal form C and amorphous form can all be used to prepare honokiol nanoliposome freeze-dried powder, and the preparation method is the same. The preparation method is given below using honokiol crystal form B as an example:

[0179] Take 50 mg of honokiol crystal form B, 500 mg of soybean lecithin, 200 mg of cholesterol, and 200 mg of Peihua phosphatidylethanolamine, dissolve them in 50 ml of anhydrous ethanol, inject them into 300 ml of purified water after complete dissolution, stir, remove ethanol by rotary evaporation, add 800 mg of sucrose as a lyophilization excipient, freeze-dry, and obtain honokiol nanoliposome lyophilized powder. After the lyophilized powder is reconstituted, the particle size of the honokiol nanoliposome is measured by a laser particle size analyzer to be 114 nm.

Claims

1. A honokiol crystal form A, characterized in that: Using Cu-Kα radiation, the honokiol crystal form A has an X-ray powder diffraction pattern as shown in FIG1 .

2. A pharmaceutical composition or pharmaceutical preparation comprising a therapeutically effective amount of the honokiol crystal form A as described in claim 1, and pharmaceutical excipients.

3. Use of the honokiol crystal form A as claimed in claim 1 in the preparation of a medicament for treating tumors.

4. Use of the honokiol crystal form A as claimed in claim 1 in the preparation of antimicrobial, antioxidant, antianxiety, antidepressant or antithrombotic drugs.

Images