阅读说明：本技术 新型parp抑制剂的稳定晶型及其制备方法 (Stable crystal form of novel PARP inhibitor and preparation method thereof ) 是由 谢军 李惠 姜春阳 彭典金 刘鹏 舒海英 许全胜 于 2021-09-09 设计创作，主要内容包括：本发明公开了新型PARP抑制剂的稳定晶型及其制备方法。所述新型PARP抑制剂为4-(3-(1,1-二氧代硫代吗啉-4-羰基)-4-氟苄基)酞嗪-1(2H)-酮,所述稳定晶型为晶型A,所述晶型A的粉末X-射线衍射图在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°的2θ处显示主峰。该晶型A纯度高,不具引湿性,具有稳定性好,粉体流动性好等特点。(The 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) phthalazin-1 (2H) -one, the stable crystal form is a crystal form A, and a powder X-ray diffraction pattern of the crystal form A shows main peaks at positions of 12.852 degrees, 13.136 degrees, 15.240 degrees, 15.528 degrees, 16.062 degrees, 17.825 degrees, 18.455 degrees, 20.003 degrees, 20.521 degrees, 20.880 degrees, 21.565 degrees, 22.894 degrees and 24.772 degrees, namely 2 theta. The crystal form A has the characteristics of high purity, no hygroscopicity, good stability, good powder flowability and the like.)

1. A stable crystalline form of a novel PARP inhibitor, characterized in that said novel PARP inhibitor is 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one, said stable crystalline form is form A, said crystalline form A having a powder X-ray diffraction pattern showing main peaks at 2 θ of 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 °, said error range of 2 θ being ± 0.2.

2. The stable crystalline form of a novel PARP inhibitor of claim 1, characterized in that said stable crystalline form a powder X-ray diffraction pattern exhibits secondary peaks at 2 Θ of 19.027 °, 19.397 °, 26.524 °,29.576 °, 34.037 °, said 2 Θ having a tolerance range of ± 0.2.

3. The stable crystalline form of a novel PARP inhibitor according to claim 1, characterized in that said stable crystalline form a powder X-ray diffraction pattern is as shown in figure 1, wherein the range of error in 2 Θ in said figure can be ± 0.2.

4. A stable crystalline form of a novel PARP inhibitor characterized in that said novel PARP inhibitor is 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one and said stable crystalline form is form B, said form B having a powder X-ray diffraction pattern showing main peaks at 2 θ of 16.439 °, 18.512 °, 20.008 °, 21.594 °, 24.070 °, 26.094 °, 27.940 °, said 2 θ having an error range of ± 0.2.

5. The preparation method of the stable crystal form A of the novel PARP inhibitor as claimed in claim 1, wherein amorphous 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one or 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one of crystal form B or C is dissolved in a first solvent under heating condition, a second solvent is added dropwise, and then the mixture is cooled and crystallized and filtered to obtain the crystal form A.

6. The method of claim 5 for preparing novel stable crystalline form A of PARP inhibitor, wherein said first solvent is selected from one or two of DMSO and DMF and said second solvent is selected from one or two of water and methyl tert-butyl ether.

7. The method of claim 6, wherein the first solvent is DMSO and the second solvent is water.

8. The preparation method of the stable crystal form B of the novel PARP inhibitor as claimed in claim 4, wherein the amorphous 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one is dissolved in the mixed solvent of dichloromethane and methanol under heating condition, and then the mixture is cooled and crystallized and filtered to obtain the PARP inhibitor.

9. Use of the stable crystalline form of a novel PARP inhibitor as defined in claim 1 or claim 4 for the preparation of an anti-tumor medicament.

10. A pharmaceutical composition prepared from the stable crystalline form of the novel PARP inhibitor of any of claims 1 or 4 and a pharmaceutically acceptable adjuvant.

Technical Field

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

Poly (ADP-ribose) polymerase (PARP) inhibitors are targeted drugs for treating tumors with Homologous Recombination Defects (HRD), which induce synthetic killing of BRCA1/2 mutated tumors by using the synthetic lethal principle, directed to specific DNA repair pathways. Currently, 4 PARP inhibitors, olapari, lucapapril, nilapapril and talapapril, 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, most among the benefits of BRCA1/2 mutant patients.

A number of novel PARP inhibitors have been disclosed in chinese patent 201810134916.4, and preliminary studies have shown that such novel PARP inhibitors, especially PARP-03, have superior tumor inhibitory effect to olaparib (AZD2281), PARP-03 being chemically named 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one, and the structural formula is as follows:

however, the product prepared by the preparation method disclosed in example 3 of chinese patent 201810134916.4 is amorphous, and has the disadvantages of instability, easy moisture absorption, and poor powder flowability, so it is necessary to develop a stable crystal form of a novel PARP-03 inhibitor.

Disclosure of Invention

The invention aims to overcome the defects in the prior art and provide a stable crystal form of a novel PARP inhibitor and a preparation method thereof.

The purpose of the invention is realized by the following technical scheme:

a stable crystalline form of a novel PARP inhibitor, characterized in that said novel PARP inhibitor is 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one, said stable crystalline form is form a, said crystalline form a having a powder X-ray diffraction pattern showing a main peak at 2 Θ of 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 ° and a secondary peak at 2 Θ of 19.027 °, 19.397 °, 26.524 °,29.576 °, 34.037 °, said 2 Θ having an error range of ± 0.2;

further, the X-ray diffraction pattern of the stable crystal form A powder is shown in the attached figure 1, and the error range of 2 theta in the attached figure can be +/-0.2.

The invention provides a stable crystal form of a novel PARP inhibitor, which is characterized in that the novel PARP inhibitor is 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -ketone, the stable crystal form is a crystal form B, a powder X-ray diffraction pattern of the crystal form B shows a main peak at 2 theta positions of 16.439 degrees, 18.512 degrees, 20.008 degrees, 21.594 degrees, 24.070 degrees, 26.094 degrees and 27.940 degrees, and shows a secondary peak at 2 theta positions of 16.874 degrees, 19.318 degrees, 20.374 degrees, 23.427 degrees, 28.037 degrees, 30.363 degrees and 32.722 degrees, and the error range of the 2 theta can be +/-0.2;

further, the X-ray diffraction pattern of the stable crystal form B powder is shown in figure 3, and the error range of 2 theta in the figure can be +/-0.2.

The invention also provides a preparation method for preparing the stable crystal form A of the novel PARP inhibitor, which is characterized in that amorphous 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one or 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one crystal form B or crystal form C is dissolved in a first solvent under the heating condition, a second solvent is dropwise added, then the temperature is reduced for crystallization, and the crystal form A is obtained by filtration;

the first solvent is one or two selected from DMSO and DMF;

the second solvent is one or two of water and methyl tert-butyl ether;

preferably, the first solvent is a combination of DMSO and the second solvent is water.

The invention also provides a preparation method for preparing the stable crystal form B of the novel PARP inhibitor, which is 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 or a mixed solvent of ethyl acetate and n-hexane under a heating condition, and then the solution is obtained by cooling, crystallizing and filtering;

preferably, the volume ratio of dichloromethane/methanol is 30: 1.

the invention also provides a novel PARP inhibitor 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazine

Application of the-1 (2H) -ketone crystal form A or the crystal form B in preparing antitumor drugs.

The invention also provides a pharmaceutical composition, which is prepared from the crystal form A or the crystal form B of 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazine-1 (2H) -one and pharmaceutic adjuvants.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a stable crystal form A and a stable crystal form B of a novel PARP inhibitor 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one, wherein the crystal form A has high purity, does not have hygroscopicity, has the characteristics of good stability, good powder flowability and the like, and is beneficial to the preparation of subsequent preparations.

Drawings

FIG. 1 is an XRPD pattern for crystalline form A of 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one;

FIG. 2 is an XRPD pattern for amorphous 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one;

FIG. 3 is an XRPD pattern for crystalline form B of 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one;

FIG. 4 is an XRPD pattern for crystalline form C of 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one;

FIG. 5 is an XRPD pattern of crystalline form B of 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one after 30 days at an elevated temperature of 60 ℃;

FIG. 6 XRPD pattern of crystalline form A of 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one prepared in example 1 after 6 months of accelerated stability;

FIG. 74 is a DSC of crystalline form A of 3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one;

FIG. 84 is a TGA diagram of crystalline form A of 3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one.

Detailed Description

The present invention will be described in detail with reference to examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that it would be apparent to those skilled in the art that several modifications and improvements can be made without departing from the inventive concept. All falling within the scope of the present invention.

Specific application examples are as follows:

EXAMPLE 14 preparation of crystalline form A of- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one

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

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, 100ml of DMF are added to a 250ml three-necked flask, stirred at room temperature for 1h, gradually precipitated solids, and subjected to LCMS follow-up reaction until the 2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoic acid is reacted completely, filtered, and the filter cake is washed with 50ml of DMF, 100ml of water, 100ml of acetone in turn. Vacuum drying at 45 deg.C to obtain 10.2g of 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one.

(2) Preparation of 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one crystal form A

And (2) heating 4.5g of the amorphous 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazine-1 (2H) -ketone obtained in the last step to 50ml of DMSO solvent, dissolving, then dropwise adding 50ml of purified water, cooling to 0-5 ℃, and filtering to obtain 3.9g of crystal form A with the HPLC purity of 99.85%. The X-ray powder diffraction pattern (XRPD pattern) obtained by Cu-Ka irradiation is shown in figure 1.

LC-MS(m/z)：416[M+H+]；

Nuclear magnetic data 1HNMR (400MHz, DMSO-d6): δ 12.58(s,1H, phthalazine proton at position 3), 8.26(d,1H, J ═ 7.7Hz, phthalazine benzen-ring proton), 7.94(d,1H, J ═ 7.9Hz, phthalazine benzen-ring proton), 7.89(t,1H, J ═ 7.4Hz, phthalazine benzen-ring proton), 7.83(t,1H, J ═ 7.4Hz, phthalazine benzen-ring proton), 7.52(d,1H, J ═ 6.3Hz, fluorobenzyl benzen-ring proton), 7.45(dd,1H, J ═ 8.0,5.4Hz, fluorobenzyl benzen-ring proton), 7.24(t,1H, J ═ 9.0Hz, fluorobenzyl proton), 4.33(s,2H, oxomorpholine cyclon proton, 3.58(s,2H, oxomorpholine, 2H, oxomorpholine, 3.27, orthomorphol proton (t, orthomorphol proton), benzylic methylene protons).

The DSC chart is shown in figure 7, and the TGA chart is shown in figure 8. As can be seen from the TGA profile, the crystalline form is free of water and solvent, and as can be seen from the DSC profile, the crystalline form melts around 287 degrees.

Example 2

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

Example 3

Amorphous 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one 3g obtained in example 1 was mixed with DMF 60ml, heated to dissolve, 600ml methyl tert-butyl ether was added dropwise, then cooled to 0-5 ℃ for crystallization, and after filtration, crystalline form A1.8 g was obtained with an HPLC purity of 99.02%.

Comparative example 1

The novel PARP inhibitor PARP-03 prepared according to the preparation method disclosed in example 3 of 201810134916.4 (i.e. step 1 of example 1) is amorphous. The XRPD pattern of amorphous 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one is shown in figure 2.

Comparative example 2

500mg of amorphous 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one obtained in example 1 is mixed with 100ml of a mixed solvent of dichloromethane/methanol (volume ratio of 30: 1), heated under reflux, cooled to 0-5 ℃ for crystallization, and filtered to obtain 250mg of crystal form B, wherein an XRPD pattern is shown in figure 3.

Comparative example 3

500mg of amorphous 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one obtained in example 1 was mixed with DMF20ml, heated to dissolve and clear, 100ml of ethyl acetate was added dropwise, and then cooled to 0-5 ℃ for crystallization, and after filtration 350mg of C form was obtained, with the XRPD pattern shown in FIG. 4.

Example 4 hygroscopicity test

A hygroscopicity experiment was performed on crystalline form A of 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one prepared in example 1, amorphous form prepared in comparative example 1, crystalline form B prepared in comparative example 2, and crystalline form C prepared in comparative example 3, as shown in Table 1.

The experimental method comprises the following steps: the hygroscopicity test is carried out according to the guiding principle of the hygroscopicity test of the Chinese pharmacopoeia 9103 medicaments. The hygroscopicity of the drug was measured at a temperature of 25. + -. 1 ℃ and a relative humidity of 80%. + -. 2%. The moisture absorption profile and definition of moisture-inducing weight gain are described as follows: has the characteristics of moisture absorption: the moisture-inducing weight is not less than 15%; moisture absorption: the moisture-drawing weight increment is less than 15% and not less than 2%; slightly hygroscopic: moisture pick-up weight less than 2% and not less than 0.2%, no or almost no moisture pick-up: the moisture-drawing weight gain is less than 0.2 percent.

Weighing about 1g of sample, placing the sample in a dryer, placing saturated sodium chloride solution at the lower part of the dryer, and calculating the weight gain percentage of the sample after placing the sample at 25-35 ℃ for 24 hours.

Table 1:

crystal form	Weight gain	Moisture-wicking property
			Amorphous form	6.8％	Has moisture absorption effect
Crystal form A	0.1％	Without hygroscopicity
			Crystal form B	1.9％	Slightly hygroscopic
Crystal form C	4.5％	Has moisture absorption effect

And (4) conclusion: the crystal form A has no hygroscopicity, and the crystal form B has slight hygroscopicity. The crystal form A is obviously superior to the crystal form C and amorphous form due to the crystal form B.

Example 5 influencing factor test

The crystal form A of 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one prepared in example 1 and the crystal form B prepared in comparative example 2 are subjected to influence factor tests such as illumination (5000 Lux/day), high temperature (60 +/-2 ℃), high humidity (75% +/-1% RH) and the like, and XRPD items of the crystal form A and the crystal form B are detected, and the results are shown in tables 2 and 3.

Table 2:

table 3:

the result shows that the investigation items of the crystal form A have no obvious change in the conditions of high humidity and illumination for 30 days, and the crystal form A is stable in the conditions of high humidity, high temperature and illumination. The crystal form B is unstable at the high temperature of 60 ℃, and part of the crystal form B is converted into the crystal form A, and an XRPD pattern is shown in an attached figure 5; form B is stable under other conditions.

Example 6 Long term stability test

The 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one crystal form A prepared in example 1 was placed in a constant temperature and humidity cabinet with the temperature of 40 ℃ plus or minus 2 ℃ and the humidity of 75% plus or minus 5% RH for 6 months, and samples were taken for analysis at 0, 1, 2, 3 and 6 months, and the appearance, total impurities (HPLC), content (HPLC), XRPD and the like were measured, and the results are shown in the following table.

Table 4:

	0 month	1 month	2 months old	3 months old	6 months old
						Appearance of the product	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	In accordance with FIG. 1	In accordance with FIG. 1	In accordance with FIG. 1	In accordance with FIG. 1	In accordance with FIG. 1
Content (wt.)	99.53％	99.41％	99.46％	99.50％	99.43％

The results show that the 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one crystal form A prepared in example 1 has no obvious change in the items tested when placed in a constant temperature and humidity cabinet with the temperature of 40 ℃ plus or minus 2 ℃ and the humidity of 75 percent plus or minus 5 percent for 6 months.

The X-ray powder diffraction pattern (XRPD) of the crystal form A prepared in example 1 after 6 months of accelerated test is shown in figure 6, and the characteristic peaks of figure 6 and figure 1 can be seen to be the same.

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

Example preparation of crystalline form A of 74- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazin-1 (2H) -one

Dissolving the 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazine-1 (2H) -ketone crystal form B (450mg) in DMSO (dimethyl sulfoxide) solvent 5ml, heating to 50-60 ℃, after clearing, dropwise adding 5ml of purified water, cooling to 0-5 ℃, and filtering to obtain the crystal form A.

Dissolving the 4- (3- (1, 1-dioxothiomorpholine-4-carbonyl) -4-fluorobenzyl) phthalazine-1 (2H) -ketone crystal form C (450mg) in DMSO (dimethyl sulfoxide) solvent 5ml, heating to 50-60 ℃, after clearing, dropwise adding 5ml of purified water, cooling to 0-5 ℃, and filtering to obtain the crystal form A.

Example 8 flowability test

According to the pharmacopoeia, the flowability of the crystal form A prepared by the method is researched through the compressibility coefficient, the bulk density and the tap density of the crystal form A and the originally-ground crystal form in the prior art are respectively measured, and then the compressibility coefficient is calculated according to the following formula. Compressibility factor (%) - (tap density-bulk density)/tap density × 100%. The results are shown in Table 5:

table 5:

crystal form	Bulk density (g/ml)	Tap density (g/ml)	Compressibility factor (%)
				Crystal form A	0.2015	0.3112	35.25％
Crystal form B	0.2102	0.3726	43.58％
				Crystal form C	0.2193	0.5904	62.85％

The result shows that the compressibility coefficient of the crystal form A is 35.25%, and the compressibility coefficient of the crystal form B is 43.58%, which shows that the crystal form A has better fluidity than the crystal form B, and is more suitable for preparation of preparations.

The foregoing description of specific embodiments of the present invention has been presented. It is to be understood that the present invention is not limited to the specific embodiments described above, and that various changes and modifications may be made by one skilled in the art within the scope of the appended claims without departing from the spirit of the invention.