Uchidelone hemihydrate single crystal and preparation method and application thereof

Abstract

To provide a crystal form of uchidelone which is used for treating solid tumors, is stable, withstands high temperature and high humidity, is prepared by various and simple methods, and is suitable for industrial production of new drugs, to provide a method for preparing the same, and to provide a pharmaceutical composition.SOLUTION: There is provided an uchidelone hemihydrate crystal which is a single crystal and has characteristic peaks at reflection angles 2 θ of about 6.1, 7.2, 12.1, 14.4, 17.2, 18.3, 18.6, 19.0, 21.3, 24.4, 29.1, and 33.4 in an X-ray powder diffraction pattern thereof.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a hemihydrate crystal of 4S,7R,8S,9S,13Z,16S-4,8-dihydroxy-5,5,7,9,13-pentamethyl-16-[E-1-methyl-2-(2-methyl-1,3-thiazol-4-yl)-prop-1-en-2-yl]-hexadecane oxygen-containing heterocycle-13-ene-2,6-ketone lactone (i.e., utidelone), a preparation method thereof, and its use in the preparation of pharmaceutical compositions, particularly for the preparation of antitumor drugs. [Background technology]

[0002] Uchiderone is a member of the epothilone family. Epothilones are naturally occurring cytotoxic compounds of the 16-membered macrolide class produced by the metabolism of the microbial myxobacterium. They share a mechanism of action similar to that of paclitaxel, which has significant antitumor activity. Both compounds induce tubulin polymerization to form a hyperstable state, inhibit microtubule depolymerization, inhibit mitosis, block tumor cell proliferation, and induce apoptosis. Epothilones exhibit potent antitumor activity in p-glycoprotein-expressing multidrug-resistant tumor cell lines and have superior water solubility compared to paclitaxel. Epothilones are superior to paclitaxel in many respects and are considered a new generation alternative to paclitaxel.

[0003] Uchideron is a member of the epothilone family and is named 4S,7R,8S,9S,13Z,16S-4,8-dihydroxy-5,5,7,9,13-pentamethyl-16-[E-1-methyl-2-(2-methyl-1,3-thiazol-4-yl)-prop-1-en-2-yl]-hexadecane oxygen-containing heterocycle-13-ene-2,6-ketone lactone, and has the following structural formula: TIFF2026004563000002.tif42170

[0004] Uchideron may be used to treat gastrointestinal tumors such as breast cancer, lung cancer, intestinal cancer, and gastric cancer; gynecological tumors such as ovarian cancer and cervical cancer; and solid tumors such as head and neck squamous cell carcinoma, esophageal cancer, pancreatic adenocarcinoma, bile duct cancer, skin cancer, brain cancer, and liver cancer. Even for the same drug, differences in crystalline form can have significant differences in hygroscopicity, stability, and bioavailability, which affect the therapeutic efficacy of the drug. Studying the crystalline form is very important for the application of utideron to the development of pharmaceutical compositions. To date, no crystalline form related to utideron has been reported. Summary of the Invention

[0005] According to one aspect of the present invention, a highly stable crystalline form of utideron is provided. The present invention provides a single crystal of utideron, which is a hemihydrate crystalline form (hemihydrate crystal, crystalline form A). The X-ray diffraction pattern is shown in Figure 1, the DSC in Figure 6, and the diffraction pattern of the single crystal in Figure 8.

[0006] Another aspect of the present invention provides a method for preparing utideron hemihydrate crystals. The method includes dissolving utideron in a solvent, allowing it to stand or stir at 2 to 25°C, and then crystallizing it. The solvent is a mixture of n-heptane and tetrahydrofuran, or a mixture of dichloromethane and n-heptane. High-purity single crystals can be obtained using this mixed solvent.

[0007] According to one specific embodiment of the present invention, the preparation method is described as follows. 3.0 mg of the initial sample of utiderone compound was weighed and placed in a 3 mL glass flask. 0.2 mL of the mixed solvent system listed in Table 1 below was added. After vortexing and appropriate ultrasonication, the solid sample was observed to be completely dissolved. The 3 mL glass flask was then sealed with a lid and allowed to stand at room temperature. After 5 days, either a needle-like white solid or a sheet-like crystalline sample precipitated in the system. XRPD detection revealed that the crystalline form of the compound prepared by this preparation method was a hemihydrate, i.e., a hemihydrate form containing two water molecules per four utiderone molecules. The X-ray powder diffraction pattern of the crystalline form A was nearly identical to that shown in Figure 1. X-ray diffraction structural analysis revealed that the crystalline utiderone hemihydrate of the present invention is a single, long, sheet-like crystal. Characterization by single crystal X-ray diffraction revealed that the crystal belongs to the triclinic P1 space group, with unit cell parameters of {a = 6.37029(4) Å, b = 14.67305(10) Å, c = 29.54548(12) Å, α = 81.3294(4) o ,β=86.3641(4) o ,γ=86.6019(5) o , V=2721.14(3) Å3}, and the Z value is shown to be 4. The details of the crystal structure of utideron hemihydrate, the result analysis and the test of refined parameters are shown in Table 2.

[0008] The crystal structure is an asymmetric structural unit, and the asymmetric structural unit of this crystal is composed of four utiderone molecules (crystallographically independent) and two water molecules, indicating that this crystal is a hemihydrate of utiderone as a compound.

[0009] The single crystal structure data successfully demonstrated the stereochemical structure of the compound utideron, and the absolute configuration of the chiral centers in the molecule is {C3(S),C6(R),C7(S),C8(S),C16(S).

[0010] As evident from DSC (differential scanning calorimetry) studies, crystalline utideron hemihydrate exhibits two characteristic DSC signals: a relatively broad endothermic signal between 50 and 110°C, which is due to the dehydrated anhydrous form of utideron hemihydrate; and a relatively sharp endothermic peak at 122±3°C, which is due to the melting of the substance (Figure 4). Crystalline utideron anhydrate exhibits only one characteristic DSC signal, a relatively sharp endothermic peak at 122±3°C, which is due to the melting of the substance (Figure 5).

[0011] The present invention relates to crystalline utideron, which is characterized by infrared (IR) spectroscopy. Measurements are carried out using 10-20 mg of utideron hemihydrate in 300 mg of potassium bromide. Characterization by IR spectroscopy in Figure 6 reveals that the utideron hemihydrate crystalline form exhibits IR (KBr) vmax values ​​of 3506, 2968, 2938, 2901, 1726, 1680, 1456, 1246, and 971 cm. -1 It has an absorption band at

[0012] The utideron hemihydrate crystalline form according to the present invention has good high temperature and humidity stability, which contributes to the retention of its specific physical properties under storage and production conditions.

[0013] Crystalline utideron has higher humidity stability than powdered utideron, and the hemihydrate crystals are very stable under high humidity conditions, with no obvious change in water content. Crystalline utideron also has higher high-temperature stability, with no change in substance even after storage at 60°C for more than one year or at 40°C for more than three years, demonstrating long-term storage stability at high temperatures.

[0014] Crystal A was determined to be the most stable crystal, and this crystal is stable for a long period under normal storage conditions and is chemically stable.

[0015] According to another aspect, the present invention provides the use of at least one crystalline form of utidellone in a pharmaceutical composition.

[0016] Based on the antitumor properties of utideron, another aspect of the present invention relates to the preparation of a pharmaceutical composition for treating diseases using utideron. The pharmaceutical composition contains crystalline utideron hemihydrate. The present invention also provides a parenterally administered injection containing the pharmaceutical composition, and an oral formulation in the form of a capsule or tablet containing 10 to 50 mg of the polycrystalline compound. An effective amount of the pharmaceutical composition of the present invention, a polycrystalline form of crystalline utideron, that has a therapeutic effect on the above mammals, particularly humans, is provided. It is preferably used to prepare a pharmaceutical composition for cancer treatment. [Brief explanation of the drawings]

[0017] [Figure 1] 1 is an X-ray powder diffraction pattern of utideron hemihydrate crystals. [Figure 2] 1 is an X-ray powder diffraction pattern of anhydrous crystalline utiderone. [Figure 3] 1 is an X-ray powder diffraction pattern of mixed crystals of utiderone. [Figure 4] FIG. 1 is a DSC diagram of crystalline utideron hemihydrate. [Figure 5] FIG. 1 is a DSC diagram of crystalline utideron anhydrate crystals. [Figure 6] 1 is an IR absorption spectrum of utideron hemihydrate. [Figure 7] 1 is an IR absorption spectrum of utideron anhydride crystals. [Figure 8] 1 is a diffraction pattern of a single crystal of a utideron compound. DETAILED DESCRIPTION OF THE INVENTION

[0018] Example The following examples are provided to further explain the crystalline utideron of the present invention and its preparation method, but the present invention is not limited to these examples. [Example]

[0019] 100 mg of utiderone was placed in a 100 mL glass flask and 6 mL of a tetrahydrofuran / n-heptane (1:5, v / v) mixed solvent was added. The mixture was vortexed and ultrasonically mixed until the sample was completely dissolved. The flask was then sealed with a lid and left at room temperature. After 5 days, a long sheet-like single crystal sample was observed. The crystals were collected, filtered under vacuum, and washed with a 30-35% ethanol solution. The crystals were then dried in a drying apparatus until the water content was reduced to 1.8-3%. X-ray diffraction pattern data (Figure 1) indicated that the resulting crystals were 0.5 hemihydrate crystals (A) with a single crystal lattice. The diffraction pattern of utiderone single crystals is shown in Figure 8. JPEG2026004563000003.jpg244170JPEG2026004563000004.jpg82170JPEG2026004563000005.jpg181170The X-ray diffraction pattern is shown in Figure 1. [Example]

[0020] 7.5 g of dried utideron solid was dissolved in 100% ethanol to a concentration of 20 mL / g, then sterilized and filtered through a 0.2 μm film. While stirring, the filtrate was gradually added with 40% water, a small amount of seed crystals, and stirred for 30 minutes. The remaining water was then added to the solution to a 50% ethanol concentration. While stirring, the temperature of the crystal solution was lowered to 4°C using a cold water bath, and stirring was continued for 2-12 hours. The crystals were filtered under vacuum, washed quickly with a cooled 30% ethanol solution at 4°C, and then dried in a vacuum dryer for 48 hours. 6 g of product was obtained. The X-ray diffraction pattern of this product is shown in Figure 2. The reflection angles 2θ of its main characteristic peaks are located at approximately 12.4, 17.5, 20.4, 21.6, 23.8, 24.8, 25.8, and 33.9, which are different from those of the hemihydrate crystals. [Example]

[0021] Determination of the high-temperature stability of utideron hemihydrate crystals. Stability test: Stability test of hemihydrate crystals was conducted under the following conditions. Storage condition 1: Stored at 2-25°C in a sealed container for more than 3 years, and NMR, HPLC, X-ray powder diffraction, and infrared spectroscopy demonstrated that the hemihydrate crystals were stable (no change in content, no generation of decomposed impurities, no change in appearance or physicochemical properties).

[0022] Storage condition 2: Store unsealed at 25°C and 60%±10% RH for at least 3 days. After storing the sample at 25°C for one year, NMR, HPLC, X-ray powder diffraction and infrared spectroscopy have demonstrated that the performance remains stable and consistent (no change in content, no generation of decomposed impurities, no change in appearance or physicochemical properties).

[0023] Storage condition 3: Stored in a sealed container at 40°C for more than two years, NMR, HPLC, X-ray powder diffraction and infrared spectroscopy have proven that all products are stable and consistent (no change in content, no generation of decomposed impurities, no change in appearance or physicochemical properties).

[0024] Storage condition 4: After storing at 60°C for more than one year, NMR, HPLC, X-ray powder diffraction and infrared spectroscopy have proven that all products are stable and consistent (no change in content, no generation of decomposition impurities, no change in appearance or physicochemical properties). [Example]

[0025] Measurement of high humidity stability and strong light stability of utideron hemihydrate crystals The hemihydrate crystal samples obtained in Example 1 were left to stand for 10 days under conditions of relative humidity of 90%±5%. Samples were taken on the 5th and 10th days, and the samples were stable, i.e., there was no obvious change in content, no decomposition impurities were generated, no changes in appearance or physicochemical properties, and the water content also showed almost no change.

[0026] In contrast, when an anhydrous crystal sample was left for 10 days under conditions of 90%±5% relative humidity and samples were taken on the 5th and 10th days, the water content increased and moisture absorption was significant.

[0027] The hemihydrate crystal sample obtained in Example 1 was placed in a light box and left for 10 days under an illumination condition of 4500 lx ± 500 lx. The sample was stable, i.e., there was no obvious change in content, no generation of decomposed impurities, no change in appearance or physicochemical properties, and the water content was also almost unchanged. [Example]

[0028] Good solubility in solubility tests of utideron hemihydrate crystals When a sample of the hemihydrate crystals obtained in Example 1 was placed in a 0.1N hydrochloric acid solution, the sample was stable, i.e., no obvious decomposition impurities were generated. The solubility was >5 mg / mL. JPEG2026004563000006.jpg48170

[0029] Approximately 25 mg of the hemihydrate crystal sample obtained in Example 1 had good stability in a self-emulsifying solution consisting of 10% ethanol, 5% propylene glycol, 45% castor oil, and 40% corn oil, with no obvious decomposition impurities observed. It could be completely dissolved in 100 mL of water, had good solubility, and no solid precipitated within 1 hour. [Example]

[0030] PK study of crystalline utideron pharmaceutical composition when administered intravenously as a hemihydrate crystal. This example fully demonstrates that crystalline utideron composition can be used to prepare pharmaceutical compositions for treating solid tumors such as breast cancer, intestinal cancer, liver cancer, gastric cancer, and lung cancer, and that its therapeutic effects are excellent.

[0031] Crystalline utideron (form A or mixed crystals containing form A) was formulated into an utideron injection solution using 50% v / v absolute ethanol (USP), 30% v / v propylene glycol (USP), and 20% v / v polyoxyethylene castor oil (Cremophor EL or ELP) as a solubilizer. The product had a low moisture content and was highly stable.

[0032] Subjects were given utideron 40 mg / m 2 The elimination half-life (t1 / 2) of utiderone in plasma after administration on days 1 and 5 was 8.6 ± 0.1 h and 8.2 ± 1.1 h, respectively, the MRT was 4.5 ± 0.8 h and 5.1 ± 0.7 h, respectively, and the AUC(0-24) after administration on days 1 and 5 was 4178.3 ± 1008.5 h·ng / mL and 4547.4 ± 1628.1 h·ng / mL, respectively. The plasma clearance (CL) was 9.2 ± 2.7 L / h / m 2 and 8.9±3.9 L / h / m 2 The apparent volume of distribution was 114.0±35.2 L / m 2 and 109.1±62.6 L / m 2 There was no statistically significant difference between the first and last doses of any parameter, so the 40 mg / m 2 After repeated administration, there was no induction of drug accumulation in the body or accelerated metabolism, indicating that there was no change in the metabolic disposition of utideron in the body.

Claims

1. Uchideron hemihydrate crystal (A) is a single crystal, and is a hemihydrate crystal type whose asymmetric structural unit is composed of four utideron molecules (crystallographically independent ones 1) and two water molecules.

2. The utideron hemihydrate crystal (A) according to claim 1, wherein the diffraction pattern of the single crystal is as shown in Figure 8.

3. Uchideron hemihydrate crystal (A) is a single crystal, and in its X-ray powder diffraction pattern, it has characteristic peaks at reflection angles 2θ of approximately 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 utideron hemihydrate crystal (A) according to claim 3, wherein the X-ray powder diffraction pattern of the single crystal is as shown in Figure 1.

5. The single crystal is a long sheet-like crystal, and the crystal belongs to the triclinic P1 space group, and the parameters of the unit cell are {a=6.37029(4) Å, b=14.67305(10) Å, c=29.54548(12) Å, α=81.3294(4)} o ,β=86.3641(4) o , γ=86.6019(5) o , V=2721.14(3) Å3} and Z value is 4. Uchideron hemihydrate crystal (A) according to any one of claims 1 to 4.

6. 6. The utideron hemihydrate crystals according to any one of claims 1 to 5, which exhibit an endothermic peak at 122±3°C at a heating rate of 10°C / min when using DSC thermal analysis.

7. A method for preparing utideron hemihydrate crystals (A) obtained using a mixed solvent of tetrahydrofuran / n-heptane 1:5 or dichloromethane / n-heptane 1:

10.

8. A pharmaceutical composition comprising the crystalline utideron hemihydrate of any one of claims 1 to 6 and a pharmaceutically acceptable excipient.

9. 9. The pharmaceutical composition according to claim 8, wherein the pharmaceutical composition is used to treat solid tumors in mammals, particularly humans.

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