Stable crystal form of novel PARP inhibitor and preparation method thereof

Abstract

The present invention discloses a stable crystal form of a novel PARP inhibitor and a preparation method thereof. The novel PARP inhibitor is 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one, and the stable crystal form is crystal form B. The powder X-ray diffraction pattern of the crystal form B shows main peaks at 2θ of 16.439°, 18.512°, 20.008°, 21.594°, 24.070°, 26.094°, and 27.940°. The crystal form B has high purity, good stability, good powder fluidity, and the like.

Description

Technical Field

[0001] The invention belongs to the field of drug synthesis and relates to a stable crystal form of a novel PARP inhibitor and a preparation method thereof. Background Art

[0002] Poly(ADP-ribose) polymerase (PARP) inhibitors are targeted drugs for the treatment of tumors with homologous recombination deficiency (HRD). They use the principle of synthetic lethality to target specific DNA repair pathways and induce synthetic killing of BRCA1 / 2 mutated tumors. Currently, four PARP inhibitors, including olaparib, rucaparib, niraparib, and talazoparib, have been approved by the U.S. Food and Drug Administration (FDA) for the treatment of ovarian cancer, metastatic breast cancer, advanced prostate cancer, and pancreatic cancer, especially in patients with BRCA1 / 2 mutations.

[0003] Chinese patent 201810134916.4 has disclosed a number of new PARP inhibitors, and preliminary studies have shown that this type of new PARP inhibitor, especially PARP-03, has a better anti-tumor effect than Olaparib (AZD2281). The chemical name of PARP-03 is 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one, and the structural formula is as follows:

[0004]

[0005] However, the product prepared according to the preparation method disclosed in Example 3 of Chinese Patent 201810134916.4 is amorphous and has disadvantages such as instability, easy moisture absorption, and poor powder fluidity. Therefore, it is necessary to develop a new stable crystalline form of PARP-03 inhibitor. Summary of the invention

[0006] The purpose of the present invention is to overcome the deficiencies of the above-mentioned prior art and to provide a new stable crystalline form of PARP inhibitor and a preparation method thereof.

[0007] The objective of the present invention is achieved through the following technical solutions:

[0008] A stable crystalline form of a novel PARP inhibitor, characterized in that the novel PARP inhibitor is 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one, the stable crystalline form is crystalline form A, and the powder X-ray diffraction pattern of crystalline form A is 12.852°, 13.136°, 15.240°, 15.528°, 16.062° , 17.825°, 18.455°, 20.003°, 20.521°, 20.880°, 21.565°, 22.894°, 24.772° 2θ show main peaks, and 19.027°, 19.397°, 26.524°, 29.576°, 34.037° 2θ show secondary peaks, and the error range of the 2θ can be ±0.2;

[0009] Furthermore, the X-ray diffraction pattern of the stable crystalline form A powder is as shown in the attached Figure 1 As shown, the error range of 2θ in the figure can be ±0.2.

[0010] The present invention provides a stable crystalline form of a novel PARP inhibitor, characterized in that the novel PARP inhibitor is 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one, the stable crystalline form is crystalline form B, the powder X-ray diffraction pattern of the crystalline form B shows main peaks at 2θ of 16.439°, 18.512°, 20.008°, 21.594°, 24.070°, 26.094°, and 27.940°, and secondary peaks at 2θ of 16.874°, 19.318°, 20.374°, 23.427°, 28.037°, 30.363°, and 32.722°, and the error range of the 2θ can be ±0.2;

[0011] Furthermore, the X-ray diffraction pattern of the stable crystalline form B powder is as shown in the attached Figure 3 As shown, the error range of 2θ in the figure can be ±0.2.

[0012] The present invention also provides a method for preparing a novel PARP inhibitor stable crystalline form A, characterized in that amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one or crystalline form B or crystalline form C of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one is dissolved in a first solvent under heating conditions, a second solvent is added dropwise, then the temperature is reduced for crystallization, and filtration is performed to obtain crystalline form A;

[0013] The first solvent is selected from one or both of DMSO and DMF;

[0014] The second solvent is selected from one or two of water and methyl tert-butyl ether;

[0015] A preferred combination is one in which the first solvent is DMSO and the second solvent is water.

[0016] The present invention also provides a method for preparing a novel PARP inhibitor stable crystalline form B, characterized in that amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one is dissolved in a dichloromethane / methanol mixed solvent or an ethyl acetate / n-hexane mixed solvent under heating conditions, then cooled to crystallize, and filtered to obtain;

[0017] Preferably, the volume ratio of dichloromethane / methanol is 30:1.

[0018] The present invention also provides a novel PARP inhibitor 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazine

[0019] Application of -1(2H)-ketone crystal form A or crystal form B in the preparation of anti-tumor drugs.

[0020] The present invention also provides a pharmaceutical composition, which is prepared from 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one crystal form A or crystal form B and pharmaceutical excipients.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The present invention provides stable crystalline form A and stable crystalline form B of a novel PARP inhibitor 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one, wherein the crystalline form A has high purity, is non-hygroscopic, has good stability, good powder fluidity, and the like, and is conducive to the preparation of subsequent preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 is an XRPD pattern of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one crystalline form A;

[0024] Figure 2 The XRPD pattern of amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one;

[0025] Figure 3 is an XRPD pattern of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one crystalline form B;

[0026] Figure 4 is an XRPD pattern of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one crystal form C;

[0027] Figure 5 This is the XRPD pattern of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one crystal form B after high temperature at 60°C for 30 days;

[0028] Figure 6 XRPD pattern of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one Form A prepared in Example 1 after 6 months of accelerated stability;

[0029] Figure 7 DSC graph of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one Form A;

[0030] Figure 8 TGA chart of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one Form A. DETAILED DESCRIPTION

[0031] The present invention is described in detail below in conjunction with embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that, for those of ordinary skill in the art, some adjustments and improvements can also be made without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0032] Specific application examples are as follows:

[0033] Example 1 Preparation of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one Crystalline Form A

[0034] (1) Preparation of amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one

[0035] To a 250ml three-necked flask, 10.0g (33.5mmol) of 2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoic acid, 4.5g (40.2mmol) of thiomorpholine 1,1-dioxide hydrochloride, 15.0g (40.2mmol) of HBTU, 6.5g (50.3mmol) of DIPEA and 100ml of DMF were added. The mixture was stirred at room temperature for 1h. Solid gradually precipitated. The reaction was monitored by LCMS until the reaction of 2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoic acid was complete. The mixture was filtered and the filter cake was rinsed with 50ml of DMF, 100ml of water and 100ml of acetone in turn. The residue was dried in vacuo at 45°C to give 10.2 g of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one.

[0036] (2) Preparation of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one Form A

[0037] The amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one obtained in the previous step was heated to 50-60°C with 50 ml of DMSO solvent, and after dissolving, 50 ml of purified water was added dropwise, and the temperature was lowered to 0-5°C. After filtering, 3.9 g of crystal form A was obtained, with an HPLC purity of 99.85%. Using Cu-Ka radiation, the obtained X-ray powder diffraction pattern (XRPD pattern) is shown in the attached figure. Figure 1 .

[0038] LC-MS (m / z): 416 [M+H+];

[0039] NMR data 1HNMR (400MHz, DMSO-d6): δ12.58 (s, 1H, phthalazine 3-position proton), 8.26 (d, 1H, J = 7.7Hz, phthalazine benzene ring proton), 7.94 (d, 1H, J = 7.9Hz, phthalazine benzene ring proton), 7.89 (t, 1H, J = 7.4Hz, phthalazine benzene ring proton), 7.83 (t, 1H, J = 7.4Hz, phthalazine benzene ring proton), 7.52 (d, 1H, J = 6.3Hz, fluorine benzyl benzene ring proton), 7.45 (dd, 1H, J = 8.0, 5.4 Hz, fluorobenzyl benzene ring proton), 7.24 (t, 1H, J = 9.0 Hz, fluorobenzyl benzene ring proton), 4.33 (s, 2H, oxomorpholine ring nitrogen ortho-proton), 3.58 (s, 2H, oxomorpholine ring nitrogen ortho-proton), 3.33 (s, 2H, morpholine ring sulfur ortho-proton), 3.27 (s, 2H, morpholine ring sulfur ortho-proton), 3.07 (s, 2H, benzyl methylene proton).

[0040] DSC graph is attached Figure 7 , TGA diagram is attached Figure 8 It can be seen from the TGA graph that the crystal form does not contain water and solvent, and it can be seen from the DSC graph that the crystal form melts at around 287 degrees.

[0041] Example 2

[0042] 1 g of amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one obtained in Example 1 was mixed with 20 ml of DMF, heated to dissolve, 10 ml of purified water was added dropwise, and then cooled to 0-5° C. for crystallization. After filtration, 500 mg of Form A was obtained with an HPLC purity of 98.81%.

[0043] Example 3

[0044] 3 g of amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one obtained in Example 1 was mixed with 60 ml of DMF, heated to dissolve, 600 ml of methyl tert-butyl ether was added dropwise, and then cooled to 0-5° C. for crystallization. After filtration, 1.8 g of Form A was obtained with an HPLC purity of 99.02%.

[0045] Comparative Example 1

[0046] According to the preparation method disclosed in Example 3 of 201810134916.4 (i.e., the method of step 1 in this Example 1), the novel PARP inhibitor PARP-03 prepared is amorphous. The XRPD pattern of amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one is shown in the attached Figure 2 .

[0047] Comparative Example 2

[0048] 500 mg of the amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one obtained in Example 1 was mixed with 100 ml of a mixed solvent of dichloromethane / methanol (volume ratio 30:1), heated to reflux, then cooled to 0-5°C for crystallization, and filtered to obtain 250 mg of Form B. The XRPD pattern is shown in the attached figure. Figure 3 .

[0049] Comparative Example 3

[0050] 500 mg of amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one obtained in Example 1 was mixed with 20 ml of DMF, heated to dissolve, 100 ml of ethyl acetate was added dropwise, and then cooled to 0-5°C for crystallization. After filtration, 350 mg of Form C was obtained. The XRPD pattern is shown in the attached figure. Figure 4 .

[0051] Example 4: Hygroscopicity Test

[0052] The 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one crystal form A prepared in Example 1, the amorphous form prepared in Comparative Example 1, the crystal form B prepared in Comparative Example 2 and the crystal form C prepared in Comparative Example 3 were subjected to a hygroscopicity test, as shown in Table 1.

[0053] Experimental method: The hygroscopicity test was carried out in accordance with the guidelines for the hygroscopicity test of drugs in Chinese Pharmacopoeia 9103. The hygroscopicity of the drug was measured at a temperature of 25±1℃ and a relative humidity of 80%±2%. The description of hygroscopic characteristics and the definition of hygroscopic weight gain are as follows: Very hygroscopic: hygroscopic weight gain is not less than 15%; Hygroscopic: hygroscopic weight gain is less than 15% and not less than 2%; Slightly hygroscopic: hygroscopic weight gain is less than 2% and not less than 0.2%; No or almost no hygroscopicity: hygroscopic weight gain is less than 0.2%.

[0054] Weigh about 1g of sample and place it in a desiccator. Place saturated sodium chloride solution at the bottom of the desiccator. After placing it at 25-35℃ for 24 hours, calculate the weight gain percentage of the sample.

[0055] Table 1:

[0056] Crystal form Weight gain Hygroscopicity Amorphous 6.8％ Hygroscopic Crystalline Form A 0.1％ No hygroscopicity Crystalline Form B 1.9％ Slightly hygroscopic Form C 4.5％ Hygroscopic

[0057] Conclusion: Form A is not hygroscopic, while Form B is slightly hygroscopic. Form A is significantly superior to Form C and amorphous due to Form B.

[0058] Example 5 Influencing Factors Test

[0059] The 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one form A prepared in Example 1 and the form B prepared in Comparative Example 2 were tested for influencing factors such as light (5000 Lux / day), high temperature (60±2°C), and high humidity (75%±1% RH), and their XRPD items were tested. The results are shown in Tables 2 and 3.

[0060] Table 2:

[0061]

[0062] Table 3:

[0063]

[0064] The results show that after 30 days of high humidity and light conditions, there is no significant change in the test items of Form A, indicating that Form A is very stable under high humidity, high temperature and light conditions. However, Form B is unstable at a high temperature of 60°C, and some of it has turned into Form A. See the attached XRPD diagram Figure 5 ; Form B is very stable under other conditions.

[0065] Example 6 Long-term stability test

[0066] The 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one crystalline form A prepared in Example 1 was placed in a constant temperature and humidity chamber at a temperature of 40°C ± 2°C and a humidity of 75% ± 5% RH for 6 months. Samples were taken for analysis at 0, 1, 2, 3, and 6 months, and its appearance, total impurities (HPLC), content (HPLC) and XRPD were tested. The results are shown in the following table.

[0067] Table 4:

[0068] 0 months 1 month 2 months 3 months 6 months Appearance Off-white powder Off-white powder Off-white powder Off-white powder Off-white powder Total Miscellaneous 0.15％ 0.13％ 0.15％ 0.16％ 0.18％ XRPD With attached Figure 1 Consistency With attached Figure 1 Consistency With attached Figure 1 Consistency With attached Figure 1 Consistency With attached Figure 1 Consistency content 99.53％ 99.41％ 99.46％ 99.50％ 99.43％

[0069] The results show that the 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazine-1(2H)-one crystalline form A prepared in Example 1 was placed in a constant temperature and humidity chamber at a temperature of 40°C±2°C and a humidity of 75%±5% for 6 months, and there was no significant change in the observed items.

[0070] The powder X-ray diffraction spectrum (XRPD) of Form A obtained in Example 1 after 6 months of accelerated testing is shown in the attached figure. Figure 6 , it can be seen Figure 6 and Figure 1 The characteristic peaks are the same.

[0071] The above studies show that the 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one crystal form A prepared in Example 1 has good stability.

[0072] Example 7 Preparation of 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one Crystalline Form A

[0073] 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one Form B (450 mg) was dissolved in 5 ml of DMSO solvent and heated to 50-60°C. After dissolving, 5 ml of purified water was added dropwise, the temperature was lowered to 0-5°C, and Form A was obtained after filtration.

[0074] 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one Form C (450 mg) was dissolved in 5 ml of DMSO solvent and heated to 50-60°C. After dissolving, 5 ml of purified water was added dropwise, the temperature was lowered to 0-5°C, and Form A was obtained after filtration.

[0075] Example 8 Fluidity Test

[0076] According to the pharmacopoeia, the fluidity of the crystal form A prepared by the present invention was studied by the compressibility coefficient, and the bulk density and tap density of the crystal form A and the original crystal form of the prior art were measured respectively, and then the compressibility coefficient was calculated according to the following formula. Compressibility coefficient (%) = (tap density-bulk density) / tap density×100%. The results are shown in Table 5:

[0077] Table 5:

[0078] Crystal form Bulk density (g / ml) Tap density(g / ml) Compressibility coefficient (%) Crystalline Form A 0.2015 0.3112 35.25％ Form B 0.2102 0.3726 43.58％ Crystalline Form C 0.2193 0.5904 62.85％

[0079] The results show that the compressibility coefficient of the crystal form A of the present invention is 35.25%, and the compressibility coefficient of the crystal form B is 43.58%, indicating that the crystal form A of the present invention has better fluidity than the crystal form B and the crystal form C is more suitable for the preparation of the preparation.

[0080] The above describes the specific embodiments of the present invention. It should be understood that the present invention is not limited to the above specific embodiments, and those skilled in the art may make various modifications or variations within the scope of the claims, which do not affect the essence of the present invention.

Claims

1. A crystalline form of a PARP inhibitor, characterized in that The PARP inhibitor is 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one, and the crystal form is crystal form B. The powder X-ray diffraction pattern of the crystal form B is shown in Figure 3.

2. A method for preparing the crystalline form B of the PARP inhibitor according to claim 1, characterized in that: Amorphous 4-(3-(1,1-dioxothiomorpholine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one is dissolved in a mixed solvent of dichloromethane and methanol under heating conditions, then cooled to crystallize, and filtered to obtain the product.

3. Use of the crystalline form of the PARP inhibitor as claimed in claim 1 in the preparation of anti-tumor drugs.

4. A pharmaceutical composition prepared from the crystal form of the PARP inhibitor according to claim 1 and pharmaceutical excipients.

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