UTIDELONE SEMI-HYDRATED SINGLE CRYSTAL AND PREPARATION METHOD THEREFOR AND USE THEREOF

Abstract

A polycrystal form of Utidelone, particularly relating to a semi-hydrated crystal form (A) of Utidelone, a preparation method therefor and a use of crystal Utidelone in preparation of a pharmaceutical composition, especially the use in preparation of a pharmaceutical composition for inhibiting tumor growth and treating solid tumors of mammals, especially human. The provided crystal form is stable and resistant to high temperature and high humidity, and the preparation method is diversified and simple, and is suitable for industrialized production of new medicines.

Claims

1. Utidelone semihydrate crystal (A), which is single crystal, and the asymmetric structure unit is composed of four Utidelone molecules (crystallographically independent 1) and two water molecules to form the semihydrate crystal form.

2. The crystal according to claim 1, wherein single crystal diffraction pattern of the crystal is shown in FIG. 8.

3. Utidelone semihydrate crystal (A) according to claim 1, which is single crystal, the X-ray powder diffraction pattern of the crystal has characteristic peaks at reflection angle 2θ of about 6.1, 7.2, 12.1, 12.7, 14.4, 15.5, 17.2, 18.3, 18.6, 19.0, 20.1, 20.4; 21.3, 23.5, 24.4, 25.6, 26.1, 29.1, and 33.4.

4. The crystal (A) according to claim 3, wherein the X-ray powder diffraction pattern of the crystal is shown in FIG. 1.

5. Utidelone semihydrate crystal (A) according to claim 1, wherein the single crystal is a long flaky crystal belonging to triclinic system, P1 space groups, and unit cell parameters are as follows: {a=6.37029(4)Å, b=14.67305(10)Å, c=29.54548(12)Å, α=81.3294(4°), β=86.3641(4°), γ=86.6019(5°), V=2721.14(3)Å3}, Z value is 4.

6. The crystal according to claim 1, wherein using DSC thermal analysis, at 10° C./min heating speed, endothermic peak appears at 122±3° C.

7. A method for preparing Utidelone semihydrate crystal (A) according to claim 1 comprising using a mixed solvent of tetrahydrofuran/n-heptane of 1:5 or dichloromethane/n-heptane of 1:10.

8. A pharmaceutical composition comprising the crystal according to claim 1 and a pharmaceutically acceptable excipient.

9. The pharmaceutical composition according to claim 8, wherein the pharmaceutical composition is used for treating a solid tumour in mammal, especially in human.

10. A method of treating for a solid tumour in mammal, especially in human, comprising administering therapeutically effective amount of the pharmaceutical composition according to claim 8.

11. The method according to claim 10, wherein the solid tumour is selected from the group consisting of breast cancer, lung cancer, colon cancer, gastrointestinal tumors such as stomach cancer, gynecological tumors such as ovarian cancer and cervical cancer, head and neck squamous cell carcinoma, esophageal cancer, pancreatic cancer, bile duct cancer, skin cancer, brain cancer and liver cancer.

Description

DESCRIPTION OF DRAWINGS

[0017] FIG. 1 is the X powder diffraction pattern of Utidelone semihydrate crystal;

[0018] FIG. 2 is the X powder diffraction pattern of Utidelone anhydrate crystal;

[0019] FIG. 3 is the X powder diffraction pattern of mixed crystals of Utidelone;

[0020] FIG. 4 is the DSC pattern of Utidelone semihydrate crystal;

[0021] FIG. 5 is the DSC pattern of Utidelone anhydrate crystal;

[0022] FIG. 6 is the IR absorption spectrum of Utidelone semihydrate crystal;

[0023] FIG. 7 is the IR absorption spectrum of Utidelone anhydrate crystal;

[0024] FIG. 8 is single crystal diffraction pattern of Utidelone compound.

EMBODIMENTS

Examples

[0025] The following examples are intended to further illustrate Utidelone crystal of the present invention and its preparation method in detail, and the content of the present invention is not limited to these examples.

Example 1

[0026] Place 100 mg of Utidelone in a 100 ml glass bottle, add 6 ml of a mixed solvent of tetrahydrofuran/n-heptane (1:5, v/v), swirl, sonicate, and mix evenly, and observe till solids were completely dissolved. The glass bottle was then sealed with a cap and placed at room temperature. After 5 days, long flaky single crystals were observed. The crystals were collected and filtered through vacuum filtration. After filtration, the crystals were washed with 30-35% ethanol solution, and then dried in a drying oven to a moisture content of 1.8-3%. According to the X-ray diffraction pattern (FIG. 1), the resulting crystal is a 0.5 semihydrate crystal (A), which is a single crystal lattice. The single crystal diffraction pattern of Utidelone is shown in FIG. 8.

TABLE-US-00001 TABLE 2 Experimental data and structural refinement parameters for the crystal structure analysis of Utidelone semihydrate Identification code 815984-03-A27 Empirical formula C.sub.27H.sub.41NO.sub.5S•0.5 H.sub.2O Formula weight 500.67 Temperature 120.00(10) K Wavelength Cu α (λ = 1.54184 Å) Crystal system, space group Triclinic, P Unit cell dimensions a = 6.37029(4) Å b = 14.67305(10) Å c = 29.54548(12) Å α = 81.3294(4)° β = 86.3641(4)° γ = 86.6019(5)° Volume 2721.14(3) Å Z, CalMolated density 4, 1.222 g/cm.sup.3 Absorption coefficient 1.363 mm.sup.−1 F(000) 1084.0 Crystal size 0.22 × 0.0214 × 0.014 mm.sup.3 2 Theta range for 6.062 to 133.184 data collection Limiting indices −7 ≤ h ≤ 7 −17 ≤ k ≤ 17 −35 ≤ l ≤ 35 Reflections collected/ 115262/18890 [R  = 0.0534, R  = 0.0322] Independent reflections Refinement method Full-matrix least-squares on F Completeness 98.0% Data/restraints/parameters 18890/3/1319 Goodness-of-fit on .sup.2 1.057 Final R indices R.sub.1 = 0.0394, wR.sub.2 = 0.1022 [  ≥ 2sigma( )] Final R indices R.sub.1 = 0.0419, wR.sub.2 = 0.1063 [all data] Largest diff. peak and hole 0.61/−0.28 e .Math. Å.sup.−3 Flack parameter −0.002(7) Bayesian statistics on Hoo  = −0.003(6), P2(true) = 1.000, Bijvo t differences P3(true) = 1.000, P3(rac-twin) = 0.0E+00, P3(false) = 0.0E+00, corr. coeff = 0.993 indicates data missing or illegible when filed

TABLE-US-00002 TABLE 3 X-ray powder diffraction results of Utidelone semihydrate Pos. Height FWHM Left d-spacing Rel. Int. [°2Th.] [cts] [°2Th.] [Å] [%] 6.078971 651.907200 0.076752 14.53936 32.71 6.389271 176.976100 0.076752 13.83392 8.88 7.207470 690.563800 0.076752 12.26521 34.65 9.222558 63.109330 0.076752 9.58935 3.17 11.693990 138.175400 0.076752 7.56768 6.93 12.133090 1992.934000 0.076752 7.29477 100.00 12.750880 243.314400 0.076752 6.94270 12.21 14.404470 228.384400 0.076752 6.14920 11.46 15.471420 182.503800 0.076752 5.72745 9.16 16.279490 128.768800 0.102336 5.44493 6.46 17.199970 1366.306000 0.076752 5.15556 68.56 17.818710 121.344500 0.076752 4.97791 6.09 18.228380 1990.086000 0.102336 4.86695 99.86 18.481630 1146.784000 0.102336 4.80082 57.54 18.999820 928.888800 0.076752 4.67104 46.61 20.109480 222.418000 0.102336 4.41572 11.16 20.351990 669.859400 0.102336 4.36365 33.61 21.242110 848.766600 0.102336 4.18277 42.59 21.673660 220.805600 0.102336 4.10045 11.08 23.488450 448.113400 0.102336 3.78759 22.49 24.375570 863.059600 0.102336 3.65172 43.31 24.785660 52.506960 0.076752 3.59222 2.63 25.591940 593.566200 0.102336 3.48084 29.78 26.124380 629.429400 0.102336 3.41110 31.58 27.379300 43.102740 0.307008 3.25754 2.16 28.279930 71.329510 0.127920 3.15581 3.58 29.084560 129.744900 0.204672 3.07031 6.51 30.518850 38.762480 0.511680 2.92920 1.94 33.350070 159.679800 0.153504 2.68672 8.01 34.857010 55.001420 0.153504 2.57395 2.76 36.455820 31.499730 0.255840 2.46465 1.58 37.080260 98.544650 0.076752 2.42457 4.94 The X-ray diffraction pattern is shown in FIG. 1.

Example 2

[0027] After dissolving 7.5 g of dry Utidelone in 100% ethanol at a concentration of 20 mL/g, sterile filtration was performed using a 0.2 urn membrane, and the filtrate was slowly added with 40 water while stirring, then adding a small amount of seed crystals, and continuing to stir for 30 minutes. The rest of the water was then added to the solution to a 50% ethanol concentration. During the stirring process, the temperature of the solution was brought to 4° C. by using a cooling water bath, and the stirring was continued for 2-12 hours. The crystals were vacuum filtered, then quickly washed with a cooled 30% aqueous ethanol solution at 4° C., and then dried in a vacuum oven for 48 hours. 6 g of product was obtained. The X-ray diffraction pattern of the product is shown in FIG. 2, and its main characteristic peaks appear at reflection angle 20 of about 12.4, 17.5, 20.4, 21.6, 23.8, 24.8, 25.8, 33.9, which is different from the semihydrate crystal.

Example 3

[0028] Utidelone Semihydrate Crystal was Determined to be Stable at High Temperature

[0029] Stability test: The stability test of the semihydrate crystal was carried out under the following conditions:

[0030] Storage condition 1: 2-25° C., stored in a sealed state for more than 3 years, confirmed by NMR, HPLC, X-ray powder diffraction and infrared spectrum analysis, etc., the semihydrate crystal was stable (no changes in content, no impurities due to degradation, no changes in appearance and physicochemical properties).

[0031] Storage Condition 2: 25° C., 60%±10% RH, unsealed storage for more than 3 days, after the sample was continued to be stored at 25° C. for 1 year, it was confirmed by NMR, HPLC, X-ray powder diffraction and infrared spectrum analysis, etc., the semihydrate crystal was stable (no changes in content, no impurities due to degradation, no changes in appearance and physicochemical properties).

[0032] Storage condition 3: Stored in a sealed state at 40° C. for more than 2 years. It was confirmed by NMR, HPLC, X-ray powder diffraction and infrared spectrum analysis, etc., the semihydrate crystal was stable (no changes in content, no impurities due to degradation, no changes in appearance and physicochemical properties).

[0033] Storage Condition 4: Stored at 60° C. for more than 1 year. It was confirmed by NMR, HPLC, X-ray powder diffraction and infrared spectrum analysis, etc., the semihydrate crystal was stable (no changes in content, no impurities due to degradation, no changes in appearance and physicochemical properties).

Example 4

[0034] Utidelone Semihydrate Crystal is Stable Under High Humidity and Strong Light

[0035] The semihydrate crystal obtained in Example 1 was placed under the condition of relative humidity of 90%±5% for 10 days, and samples were taken on the 5th and 10th days. The samples were stable: no obvious changes in content, no impurities due to degradation, no changes in appearance and physicochemical properties, and the moisture content was basically unchanged.

[0036] Utidelone anhydrous crystal was placed under the condition of relative humidity of 90±5% for 10 days. Samples were taken on the 5th and 10th days. The moisture content increased and the moisture absorption was obvious.

[0037] The semihydrate crystal obtained in Example 1 was placed in a light box, and placed for 10 days under the condition that the light intensity was 4500 lx±500 lx. The crystal was stable: no obvious changes in content, no impurities due to degradation, no changes in appearance and physicochemical properties, and the moisture content was basically unchanged.

Example 5

[0038] Utidelone Semihydrate Crystal Showed Good Solubility in a Solubility Test

[0039] The semihydrate crystal obtained in Example 1 was placed in a 0.1N hydrochloric acid solution, and the crystal was stable: no obvious degradation impurities were produced. Solubility >5 mg/ml.

TABLE-US-00003 Solvent Solubility Ethanol >50 mg/ml Methanol >50 mg/ml Propylene Glycol >50 mg/ml DMSO >50 mg/ml Benzyl alcohol >50 mg/ml 50% Ethanol, 30% Propylene Glycol, 20% Castor Oil >100 mg/ml 

[0040] About 25 mg of the semihydrate crystal obtained in Example 1 was formed self-emulsifying solution in 10% ethanol, 5% propylene glycol, 45% castor oil, and 40% corn oil. The self-emulsifying solution has good stability and no obvious degradation impurities were observed. It was able to be completely dissolved in 100 ml of water, showing good solubility and no solid was precipitated within 1 hour.

Example 6

[0041] PK Study on Intravenous Administration of the Pharmaceutical Composition Containing Utidelone Semihydrate Crystal

[0042] This example fully illustrates that the composition may be used to prepare a medicament for treating solid tumors such as breast cancer, intestinal cancer, liver cancer, stomach cancer, lung cancer, etc., and its curative effect is excellent.

[0043] Utidelone injection was prepared with Utidelone crystals (crystal A or mixed crystals containing crystal A) and 50% v/v absolute ethanol (USP), 30% v/v propylene glycol (USP) and 20% v/v polyoxyl castor oil 1 (Cremophor EL or ELP) as a solubilizer. The injection has low water content and high stability.

[0044] Subjects were given intravenous infusion of Utidelone 40 mg/m2, once a day, for 5 consecutive days, the elimination half-life t½ of Utidelone in plasma after administration on the 1st and 5th days were respectively 8.6±0.1 h and 8.2±1.1 h, MRT were respectively 4.5±0.8 h and 5.1±0.7 h, AUC(0-24) were respectively 4178.3±1008.5 h.Math.ng/mL and 4547.4±1628.1 h.Math.ng/mL; plasma clearance CL were 9.2±2.7 L/h/m2 and 8.9±3.9 L/h/m2, respectively; apparent volume of distribution were 114.0±35.2 L/m2 and 109.1±62.6 L/m2, respectively. The difference between the first time and the last time of all parameters was not statistically significant, indicating that after continuous administration of 40 mg/m2, the drug did not accumulate and induce accelerated metabolism in the body, and thus the metabolic disposal of Utidelone in the body did not change.