Taxol antitumor compound

Abstract

The invention provides a paclitaxel antitumor compound. Specifically, the invention provides a hydrate and a crystal form of 2 '-acetyl docetaxel. The hydrate or the preferred crystal form A of 2 '-acetyl docetaxel is superior to other crystal forms or amorphous forms in the aspects of solid stability, hygroscopicity and mechanical force stability, and has better druggability and clinical application value.

Description

Taxol antitumor compounds Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical chemistry, and in particular relates to a paclitaxel anti-tumor compound and a preparation method and application thereof. Background Art

[0002] Docetaxel is a semi-synthetic product of 10-deacetylbaccatin III, a non-cytotoxic precursor extracted from the needles of Taxus chinensis. It is a paclitaxel analogue that acts on cellular microtubules, promoting tubulin polymerization while inhibiting microtubule depolymerization, causing cells to arrest in the G2 / M phase and inhibiting mitosis, thereby killing tumor cells. It has stronger anti-tumor activity than paclitaxel. 2'-Acetyl docetaxel (CAS: 151509-27-2), a synthetic intermediate or by-product of docetaxel, was first published in a journal article (Tetrahedron (1993), 49(30), 6533-44, by Dubois, Joelle, et al.). Its structure is shown below.

[0003] Summary of the Invention

[0004] The inventors of this application conducted the first systematic study on 2'-acetyldocetaxel as a drug. After extensive experiments and screening, they finally obtained a drug substance with stable quality, low hygroscopicity, and suitable for drug formulation.

[0005] Therefore, in one aspect, the present invention provides a hydrate of 2'-acetyldocetaxel:

[0006] Here, n is 0.5-3, preferably 1-2.

[0007] Preferably, the hydrate is a sesquihydrate.

[0008] In another aspect, the present invention provides a crystalline form A of 2'-acetyldocetaxel, which has characteristic peaks at 6.0±0.2°, 9.8±0.2°, 10.4±0.2°, 13.6±0.2°, and 19.0±0.2° in X-ray powder diffraction using Cu-Kα radiation and expressed in 2θ angles.

[0009] Preferably, the crystalline form A uses Cu-Kα radiation, and the X-ray powder diffraction expressed in 2θ angles has characteristic peaks at 6.0±0.2°, 9.8±0.2°, 10.4±0.2°, 13.6±0.2°, 15.2±0.2°, 16.2±0.2°, and 19.0±0.2°.

[0010] Also preferably, the crystalline form A uses Cu-Kα radiation, and the X-ray powder diffraction expressed in 2θ angles has characteristic peaks at 6.0±0.2°, 9.8±0.2°, 10.4±0.2°, 11.4±0.2°, 12.4±0.2°, 13.6±0.2°, 15.2±0.2°, 16.2±0.2°, and 19.0±0.2°.

[0011] Further preferably, the crystalline form A has an X-ray powder diffraction spectrum substantially as shown in FIG3 .

[0012] In some embodiments of the present invention, the differential scanning calorimetry curve of the crystalline form A has an endothermic peak starting point at 169.0±5°C.

[0013] In some embodiments of the present invention, the crystalline form A has a DSC spectrum substantially as shown in FIG4 .

[0014] In some embodiments of the present invention, the crystalline form A has a TGA spectrum substantially as shown in FIG5 .

[0015] According to the present invention, the crystalline form A of 2'-acetyldocetaxel is preferably a hydrate, more preferably a sesquihydrate.

[0016] On the other hand, the present invention also provides a crystalline form B of 2'-acetyldocetaxel, which uses Cu-Kα radiation and has characteristic peaks in X-ray powder diffraction expressed in 2θ angles at 5.9±0.2°, 10.0±0.2°, 10.4±0.2°, 13.6±0.2°, and 18.9±0.2°.

[0017] Preferably, the crystalline form B uses Cu-Kα radiation, and the X-ray powder diffraction expressed in 2θ angles has characteristic peaks at 5.9±0.2°, 10.0±0.2°, 10.4±0.2°, 13.6±0.2°, 16.1±0.2°, 18.9±0.2°, and 19.6±0.2°.

[0018] Further preferably, the crystalline form B has an X-ray powder diffraction spectrum substantially as shown in FIG8 .

[0019] In yet another aspect, the present invention further provides a pharmaceutical composition comprising the aforementioned crystalline form or hydrate of 2'-acetyldocetaxel.

[0020] Preferably, the pharmaceutical composition of the present invention comprises the aforementioned crystalline form or hydrate of 2'-acetyldocetaxel and pharmaceutically acceptable excipients.

[0021] In yet another aspect, the present invention further provides use of a crystalline form or hydrate of 2'-acetyl docetaxel in the preparation of a medicament for treating a tumor disease. In other embodiments, the present invention provides a crystalline form or hydrate of 2'-acetyl docetaxel for use in treating a tumor disease. In other embodiments, the present invention provides a method for treating a tumor disease, comprising administering a therapeutically effective amount of a crystalline form or hydrate of 2'-acetyl docetaxel.

[0022] Those skilled in the art will readily appreciate that the compounds disclosed herein and their crystalline forms can be mixed with pharmaceutically acceptable excipients well known in the art. Specifically, as a drug for systemic administration, it can be formulated into a dosage form suitable for oral or parenteral administration.

[0023] To treat neoplastic diseases, crystalline forms of 2'-acetyl docetaxel can be co-administered with other therapeutic agents. Therefore, another aspect of the present invention provides a combination pharmaceutical composition for treating neoplastic diseases, comprising a therapeutically effective amount of a crystalline form of 2'-acetyl docetaxel and one or more other therapeutic agents. To treat neoplastic diseases, crystalline forms of 2'-acetyl docetaxel can be co-administered with other therapeutic modalities, including but not limited to radiotherapy. Beneficial effects

[0024] The present invention provides a hydrate and crystalline form of 2'-acetyldocetaxel. The inventors unexpectedly discovered that the hydrate or preferably crystalline form A of 2'-acetyldocetaxel possesses excellent properties, such as stable physicochemical stability, extremely low hygroscopicity, and superior mechanical stability. In particular, it surpasses other crystalline forms or amorphous forms in terms of hygroscopicity and stability, demonstrating improved drugability and clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] FIG1 is an XRPD spectrum of an amorphous form of 2'-acetyldocetaxel.

[0026] FIG2 is a DVS graph of an amorphous form of 2'-acetyldocetaxel.

[0027] FIG3 is an XRPD spectrum of 2'-acetyldocetaxel Form A.

[0028] FIG4 is a DSC spectrum of 2'-acetyldocetaxel Form A.

[0029] FIG5 is a TGA spectrum of 2'-acetyldocetaxel Form A.

[0030] FIG6 is a schematic diagram of the asymmetric unit of the single crystal structure of 2'-acetyldocetaxel Form A.

[0031] FIG7 is an overlay of the XRPD of the single crystal structure and the XRPD of Form A of the compound starting material.

[0032] FIG8 is an XRPD spectrum of 2'-acetyldocetaxel Form B.

[0033] FIG9 is a DVS diagram of Form A.

[0034] FIG10 is an XRPD pattern of Form A before and after DVS testing.

[0035] FIG11 is an XRPD pattern of Form A before and after tableting and manual grinding. DETAILED DESCRIPTION

[0036] The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. The following embodiments are intended only to illustrate and explain the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are encompassed within the scope of protection that the present invention is intended to protect.

[0037] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0038] DSC and TGA testing instruments: TA Discovery 5500 TGA Thermogravimetric Analyzer and TA Discovery 2500 DSC Differential Scanning Calorimeter. Measurement parameters are as follows:

[0039] XRPD test instrument name and model: PANalytical X'Pert3 and Empyrean X-ray powder diffraction analyzer. Test parameters are as follows:

[0040] Dynamic moisture sorption (DVS) curves were collected on a DVS Intrinsic SMS (Surface Measurement Systems). Relative humidity at 25°C was calibrated using the deliquescent points of LiCl, Mg(NO3)2, and KCl. The test parameters are as follows:

[0041] Single crystal diffraction instrument name and model: Bruker D8 VENTURE single crystal diffractometer. Test conditions are as follows:

[0042] Example 1. Preparation of 2'-acetyl docetaxel

[0043] 2'-Acetyl docetaxel was obtained according to the method of steps 1-3 in Example 1 of prior art patent CN102050804A.

[0044] 1H-NMR 600MHz (CDCl3): δ1.119 (s, 3H), 1.227 (s, 3H), 1.332 (s, 9H), 1.744 (s, 3H), 1.831-1.874 (m, 1H), 1.935 (s, 3H), 2.078 (s, 3H), 2.152-2.328(m, 2H), 2.433(s, 3H), 2.550-2.600(m, 1H), 3.919-3.929(m, 1H), 4.190-4.204(m, 1H), 4.252-4.281(m, 1H), 4.312-4.327(m, 1H), 4.955-4.970(m, 1H), 5.219(s, 1H), 5.366(s, 1H), 5.477-5.490(m, 2H), 5.676-5.687(m, 1H), 6. MS m / z 850.45([M+H]+).

[0045] The obtained 2'-acetyl docetaxel was detected to be an amorphous form, and the XRPD analysis results are shown in Figure 1. Figure 2 shows the DVS pattern of the amorphous form. As shown in Figure 2, the amorphous sample absorbed moisture at 25°C / 80% RH and gained 4.0% to 4.6% in weight, indicating hygroscopicity. The XRPD results after DVS analysis revealed faint diffraction peaks (consistent with Form A), indicating a trend of transformation from the amorphous form to Form A.

[0046] Examples 2 to 5. Preparation of 2'-acetyl docetaxel Form A

[0047] Approximately 20 mg of 2'-acetyl docetaxel was weighed into a 3 mL vial, 1.0-2.0 mL of the solvent shown below was added, and the mixture was stirred at 50°C overnight. The mixture was then filtered (through a PTFE filter with a pore size of 0.45 μm) to obtain the filtrate. The filtrate was placed in a biochemical incubator and cooled from 50°C to 5°C at a cooling rate of 0.1°C / min. After slow cooling, Form A was obtained by XRPD, TGA, and DSC analysis.

[0048] The XRPD test results of the crystal form A are shown in Figure 3, the XRPD diffraction peak data are listed in Table 1, and the DSC and TGA test results are shown in Figures 4 and 5. The DSC results of this crystal form show that there is an endothermic peak at 169.0°C (onset temperature).

[0049] Table 1: XRPD diffraction peak data of Form A

[0050] It is noteworthy that when the same method as in Example 2-5 was used, except that the solvent was replaced with methyl tert-butyl ether or tetrahydrofuran / water (volume ratio 1:1), the amorphous form of 2'-acetyldocetaxel was obtained.

[0051] From the plate-like crystal sample obtained by culturing Form A obtained in the above example, a single crystal with suitable diffraction quality was cut and selected, and single crystal X-ray diffraction was performed on it to characterize its crystal structure. The single crystal X-ray diffraction characterization results showed that the crystal belonged to the orthorhombic system, the P212121 space group, and its unit cell parameters were: { α=90°, β=90°, γ=90°, Figure 6 is a schematic diagram of the asymmetric unit of the single crystal structure of Form A. The asymmetric unit of this crystal structure contains one molecule and a total of approximately 1.5 molecules of crystalline water, indicating that the single crystal is a hydrate. Figure 7 is an overlay of the calculated XRPD of the single crystal structure and the XRPD of Form A, the compound starting material, which are consistent.

[0052] Example 6. Preparation of 2'-acetyl docetaxel Form B

[0053] Form A obtained in Example 2 was heated to 150°C under N2 protection to obtain Form B. The XRPD results of Form B are shown in Figure 8. Maintaining N2 protection and cooling to 30°C, Form B remained unchanged. Form B was exposed to room temperature (~20% RH) and immediately transformed back into Form A. The obtained Form B absorbed water under room temperature and transformed back into Form A, becoming unstable.

[0054] Test Example 1. Solid-state stability

[0055] An appropriate amount of Form A sample was weighed and stability experiments were performed under the conditions of 60°C / closed / 1 day, 25°C / 60% RH / open / 1 week, and 40°C / 75% RH / open / 1 week. The solid samples under different conditions were evaluated for physical stability by XRPD test of crystal form, and for chemical stability by UPLC test of purity. The results of the physical stability evaluation of Form A are summarized in Table 2, and the results of the chemical stability evaluation are summarized in Table 3. The solid-state stability results showed that Form A did not undergo crystal form transformation or a significant decrease in UPLC purity after being placed under the evaluation conditions, indicating that Form A has good physical and chemical stability under the evaluation conditions.

[0056] Table 2 Summary of solid-state stability evaluation results of Form A

[0057] Table 3 UPLC results of solid-state stability evaluation of Form A

[0058] Test Example 2. Hygroscopicity

[0059] The hygroscopicity of Form A was evaluated by a dynamic moisture sorption test (DVS) at 25°C between 0% RH and 95% RH. The DVS results and the XRPD comparison results of the tested samples are shown in Figures 9 and 10. The evaluation results show that Form A has a moisture absorption weight gain of 1.8% under 25°C / 80% RH conditions, indicating that it is only slightly hygroscopic, which is significantly lower than the hygroscopicity of the amorphous form (4.0% to 4.6%) measured under the same conditions in Example 1. In addition, no obvious plateau in water absorption was observed during the adsorption and desorption process. The XRPD results show that the crystalline form of Form A did not undergo any transformation before and after the DVS test.

[0060] Test Example 3. Mechanical Stability

[0061] To evaluate the mechanical stability of Form A, Form A was compressed (350 MPa) and manually ground. XRPD analysis was performed on the compressed and manually ground samples. As shown in Figure 11, Form A did not undergo polymorphic transformation after compression at 350 MPa and after manual grinding for 3 minutes.

[0062] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of protection of the present invention.

Claims

1. A hydrate of 2'-acetyldocetaxel: in, n is 0.5-3, preferably 1-2.

2. The hydrate of 2'-acetyldocetaxel according to claim 1, wherein The hydrate is a sesquihydrate.

3. A crystalline form A of 2'-acetyldocetaxel, wherein: Using Cu-Kα radiation, X-ray powder diffraction expressed in 2θ angles has characteristic peaks at 6.0±0.2°, 9.8±0.2°, 10.4±0.2°, 13.6±0.2°, and 19.0±0.2°.

4. The crystal form according to claim 3, wherein The crystal form uses Cu-Kα radiation, and X-ray powder diffraction expressed in 2θ angles has characteristic peaks at 6.0±0.2°, 9.8±0.2°, 10.4±0.2°, 13.6±0.2°, 15.2±0.2°, 16.2±0.2°, and 19.0±0.2°.

5. The crystal form according to claim 3, wherein The crystal form uses Cu-Kα radiation, and the X-ray powder diffraction expressed in 2θ angles has characteristic peaks at 6.0±0.2°, 9.8±0.2°, 10.4±0.2°, 11.4±0.2°, 12.4±0.2°, 13.6±0.2°, 15.2±0.2°, 16.2±0.2°, and 19.0±0.2°.

6. The crystalline form according to any one of claims 3 to 5, wherein The crystalline form has an X-ray powder diffraction pattern substantially as shown in FIG3 .

7. The crystalline form according to any one of claims 3 to 6, wherein The differential scanning calorimetry curve of the crystal form has an endothermic peak starting point at 169.0±5°C.

8. The crystalline form according to any one of claims 3 to 7, wherein The crystalline form has a DSC spectrum substantially as shown in FIG4 .

9. The crystalline form according to any one of claims 3 to 8, wherein The crystalline form has a TGA spectrum substantially as shown in FIG5 .

10. The crystalline form according to any one of claims 3 to 9, wherein The crystal form is a hydrate, preferably a sesquihydrate.

11. A pharmaceutical composition comprising the hydrate according to any one of claims 1-2 or the crystal form according to any one of claims 3-10.

Images