Crystal form of bicyclic derivative PARP inhibitor as well as preparation method and application of crystal form

Abstract

The invention provides a compound represented by formula (I), a crystal form, a preparation method and a composition thereof, and applications of the compound and the crystal form in preparation of related drugs. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to multiple crystal forms of a compound, and a preparation method and application thereof, in particular to multiple crystal forms of a bicyclic derivative PARP inhibitor, and a preparation method and application thereof, belonging to the technical field of medicinal chemistry. Background Art

[0002] Approximately 5% of breast cancer patients are associated with germline mutations in the BRCA1 / 2 genes (3% for the BRCA1 gene and 2% for the BRCA2 gene). Most breast cancers caused by BRCA1 mutations are triple-negative breast cancers (70%), while BRCA2 mutations are more likely to cause estrogen receptor-positive breast cancers (70%). The BRCA1 / 2 genes are tumor suppressor genes and play important roles in DNA damage repair, normal cell growth, etc. Mutations in these genes can inhibit the normal repair ability after DNA damage, causing homologous recombination deficiency (HRD), that is, the loss of BRCA function or mutations or loss of function in other homologous recombination-related genes, resulting in the inability of double-stranded broken DNA to be repaired by homologous recombinant repair (HRR), and ultimately leading to canceration.

[0003] Poly(ADP-ribose) polymerase (PARP) is a DNA repair enzyme and plays a key role in the DNA repair pathway. When DNA is damaged and broken, PARP is activated. As a molecular sensor of DNA damage, it has the function of recognizing and binding to the DNA break position, and then activates and catalyzes the poly-ADP-ribosylation of the receptor protein, participating in the DNA repair process. PARP plays a key role in the excision and repair of single-stranded DNA bases. In HRD tumor cells, DNA double strands cannot be repaired, and PARP inhibitors block single-strand repair, thus forming a "synthetic lethality" effect, resulting in the death of tumor cells.

[0004] PARP inhibitors have a "trapping" effect on PARP proteins, causing the PARP proteins bound to damaged DNA to be trapped on the DNA and unable to come down, directly resulting in the inability of other DNA repair proteins to bind, ultimately leading to cell death. Currently, several PARP inhibitors have been successfully developed, such as olaparib, rucaparib, and niraparib, etc. However, adverse reactions limit their ability to be combined with chemotherapy drugs. This may be related to the lack of selectivity for the PARP family in the marketed PARP inhibitors. These side effects include intestinal toxicity caused by telomere end transferase inhibition and hematotoxicity caused by PARP-2 inhibition. Therefore, developing highly selective PARP-1 inhibitors and reducing the related toxic side effects of non-selective PARP inhibitors has important clinical significance.

[0005] Compound (I) described in WO2023227052 has good PARP inhibitory activity.

[0006]

[0007] It is very important to select a stable, reproducibly manufacturable, and crystalline form with physicochemical properties favorable for its use as a therapeutic agent. SUMMARY OF THE INVENTION

[0008] The present invention provides a compound of formula (I), its crystal forms and preparation methods, pharmaceutical compositions, and their use in the preparation of drugs for treating PARP-mediated diseases. The various crystal forms provided by the present invention have excellent properties such as high purity, good solubility, stable physical and chemical properties, easy to process and crystallize, handle, resistant to high temperature, high humidity and strong light, and low hygroscopicity.

[0009] The present invention provides a compound of formula (I):

[0010]

[0011] Furthermore, the compound of formula (I) is a crystal form.

[0012] In certain specific embodiments, the crystal form is Crystal Form I. Using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 9.17° ± 0.2°, 10.37° ± 0.2°, 13.84° ± 0.2°, 15.80° ± 0.2°, 16.24° ± 0.2°, 17.05° ± 0.2°, and 17.92° ± 0.2°.

[0013] In certain specific embodiments, the crystal form is Crystal Form I. Using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 9.17° ± 0.2°, 10.37° ± 0.2°, 13.84° ± 0.2°, 16.24° ± 0.2°, 17.05° ± 0.2°, 17.92° ± 0.2°, 18.45° ± 0.2°, 20.41° ± 0.2°, 21.47° ± 0.2°, 22.18° ± 0.2°, 22.47° ± 0.2°, 24.34° ± 0.2°, 24.95° ± 0.2°, 25.24° ± 0.2°, and 25.97° ± 0.2°.

[0014] In certain specific embodiments, the crystal form is Crystal Form I. Using Cu-Kα radiation, its X-ray powder diffraction pattern is substantially as Figure 2 shown.

[0015] In a specific embodiment, the differential scanning calorimetry (DSC) curve of polymorph I shows that its peak temperature is 295.11 °C and ΔH = 102.89 J / g.

[0016] In certain specific embodiments, polymorph I of the compound of formula (I), its differential scanning calorimetry (DSC) curve is substantially as Figure 3 shown.

[0017] In certain specific embodiments, polymorph I of the compound of formula (I), its thermogravimetric analysis (TGA) curve shows a weight loss of about 0.12% before 100 °C and a weight loss of about 0.68% from 100 °C to 300 °C.

[0018] In certain specific embodiments, polymorph I of the compound of formula (I), its thermogravimetric analysis curve is substantially as Figure 4 shown.

[0019] In a specific embodiment, the isothermal adsorption curve (DVS) of polymorph I shows that at 80% humidity, the moisture absorption weight gain of this polymorph is 0.4318%.

[0020] In certain specific embodiments, polymorph I of the compound of formula (I), its isothermal adsorption curve is substantially as Figure 5 shown.

[0021] In certain specific embodiments, the polymorph is polymorph II. Using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 7.49° ± 0.2°, 11.23° ± 0.2°, 14.76° ± 0.2°, 17.63° ± 0.2°, 18.77° ± 0.2°, 21.43° ± 0.2°, 21.71° ± 0.2° and 24.32° ± 0.2°.

[0022] In certain specific embodiments, the polymorph is polymorph II. Using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 7.49° ± 0.2°, 11.23° ± 0.2°, 11.83° ± 0.2°, 14.76° ± 0.2°, 17.63° ± 0.2°, 18.77° ± 0.2°, 21.43° ± 0.2°, 21.71° ± 0.2°, 22.39° ± 0.2°, 24.32° ± 0.2°, 25.34° ± 0.2° and 25.72° ± 0.2°.

[0023] In certain specific embodiments, for polymorph II using Cu-Kα radiation, its X-ray powder diffraction pattern is substantially as Figure 6 shown.

[0024] In a specific embodiment, the differential scanning calorimetry curve (DSC) of polymorph II shows that its peak temperature is 295.15 °C and ΔH = 89.910 J / g.

[0025] In certain specific embodiments, polymorph II of the compound of formula (I), whose differential scanning calorimetry curve is substantially as Figure 7 shown.

[0026] In a specific embodiment, polymorph II of the compound of formula (I), whose thermogravimetric analysis curve (TGA) shows a weight loss of about 0.08% before 100 °C and a weight loss of about 0.97% from 100 °C to 300 °C.

[0027] In a specific embodiment, polymorph II of the compound of formula (I), the thermogravimetric analysis curve is substantially as Figure 8 shown.

[0028] In a specific embodiment, the isothermal adsorption curve (DVS) of polymorph II shows that at 80% humidity, the moisture absorption weight gain of this polymorph is 0.726%.

[0029] In certain specific embodiments, polymorph II of the compound of formula (I), the isothermal adsorption curve is substantially as Figure 9 shown.

[0030] The present invention also provides a pharmaceutical composition, wherein the pharmaceutical composition contains a therapeutically effective amount of the aforementioned compound or any polymorph, as well as a pharmaceutically acceptable carrier and / or excipient. Preferably, the therapeutically effective amount is 1 - 1440 mg. The pharmaceutical composition can be in the form of unit dosage forms (unit dosage forms are also referred to as "formulation specifications").

[0031] The present invention also provides the use of the compound, polymorph or composition according to any one of the aforementioned aspects in the preparation of a drug for treating PARP-mediated diseases. Further, the PARP-mediated diseases are selected from breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer.

[0032] The present invention also provides a method for treating PARP-mediated diseases, the method comprising administering to a subject a therapeutically effective amount of the compound, polymorph or its composition according to any one of the aforementioned aspects. The disease is preferably breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer. Preferably, the therapeutically effective amount is 1 - 1440 mg. In some embodiments, the mammals in the present invention include humans.

[0033] As used herein, "effective amount" or "therapeutically effective amount" means an amount of the crystalline form disclosed herein that, to some extent, alleviates one or more symptoms of the disease or disorder being treated. In some embodiments, the result is a reduction and / or alleviation of the signs, symptoms, or causes of the disease, or any other desired alteration of a biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising a compound or crystalline form disclosed herein that is required to provide a clinically significant reduction in the symptoms of the disease.Examples of a therapeutically effective amount include, but are not limited to, 1 - 1440 mg, 1 - 1400 mg, 1 - 1300 mg, 1 - 1200 mg, 1 - 1000 mg, 1 - 900 mg, 1 - 800 mg, 1 - 700 mg, 1 - 600 mg, 1 - 500 mg, 1 - 400 mg, 1 - 300 mg, 1 - 250 mg, 1 - 200 mg, 1 - 150 mg, 1 - 125 mg, 1 - 100 mg, 1 - 80 mg, 1 - 60 mg, 1 - 50 mg, 1 - 40 mg, 1 - 25 mg, 1 - 20 mg, 5 - 1000 mg, 5 - 900 mg, 5 - 800 mg, 5 - 700 mg, 5 - 600 mg, 5 - 500 mg, 5 - 400 mg, 5 - 300 mg, 5 - 250 mg, 5 - 200 mg, 5 - 150 mg, 5 - 125 mg, 5 - 100 mg, 5 - 90 mg, 5 - 70 mg, 5 - 80 mg, 5 - 60 mg, 5 - 50 mg, 5 - 40 mg, 5 - 30 mg, 5 - 25 mg, 5 - 20 mg, 10 - 1000 mg, 10 - 900 mg, 10 - 800 mg, 10 - 700 mg, 10 - 600 mg, 10 - 500 mg, 10 - 450 mg, 10 - 400 mg, 10 - 300 mg, 10 - 250 mg, 10 - 200 mg, 10 - 150 mg, 10 - 125 mg, 10 - 100 mg, 10 - 90 mg, 10 - 80 mg, 10 - 70 mg, 10 - 60 mg, 10 - 50 mg, 10 - 40 mg, 10 - 30 mg, 10 - 20 mg; 20 - 1000 mg, 20 - 900 mg, 20 - 800 mg, 20 - 700 mg, 20 - 600 mg, 20 - 500 mg, 20 - 400 mg, 20 - 350 mg, 20 - 300 mg, 20 - 250 mg, 20 - 200 mg, 20 - 150 mg, 20 - 125 mg, 20 - 100 mg, 20 - 90 mg, 20 - 80 mg, 20 - 70 mg, 20 - 60 mg, 20 - 50 mg, 20 - 40 mg, 20 - 30 mg; 50 - 1000 mg, 50 - 900 mg, 50 - 800 mg, 50 - 700 mg, 50 - 600 mg, 50 - 500 mg, 50 - 400 mg, 50 - 300 mg, 50 - 250 mg, 50 - 200 mg, 50 - 150 mg, 50 - 125 mg, 50 - 100 mg; 100 - 1000 mg, 100 - 900 mg, 100 - 800 mg, 100 - 700 mg, 100 - 600 mg, 100 - 500 mg, 100 - 400 mg, 100 - 300 mg, 100 - 250 mg, 100 - 200 mg;.

[0034] In some embodiments, the pharmaceutical composition or formulation of the present invention contains the above-mentioned therapeutically effective amount of the compound or crystal form of the present invention;

[0035] The present invention relates to a pharmaceutical composition or pharmaceutical formulation, and the pharmaceutical composition or pharmaceutical formulation contains a therapeutically effective amount of the compound, crystal form of the present invention, as well as a carrier and / or excipient. The pharmaceutical composition can be in the form of a unit dosage form (the amount of the active ingredient in the unit dosage form is also referred to as the "formulation specification"). In some embodiments, the pharmaceutical composition includes, but is not limited to, 1 - 1440 mg, 5 - 1000 mg, 10 - 800 mg, 20 - 600 mg, 25 - 500 mg, 40 - 200 mg, 50 - 100 mg, 1 mg, 1.25 mg, 2.5 mg, 5 mg, 10 mg, 12.5 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1440 mg of the crystal form of the present invention.

[0036] A method for treating a disease in a mammal, the method comprising administering to a subject a therapeutically effective amount of the compound or crystal form of the present invention, as well as a pharmaceutically acceptable carrier and / or excipient, the therapeutically effective amount being preferably 1 - 1440 mg, and the disease being selected from breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer.

[0037] A method for treating a disease in a mammal, the method comprising administering to a subject a daily dose of 1-1440 mg / day of a medicament, the inventive compound or crystalline form, and a pharmaceutically acceptable carrier and / or excipient, the daily dose may be a single dose or divided doses. In some embodiments, the daily dose includes, but is not limited to, 10-1440 mg / day, 20-1440 mg / day, 25-1440 mg / day, 50-1440 mg / day, 75-1440 mg / day, 100-1440 mg / day, 200-1440 mg / day, 10-1000 mg / day, 20-1000 mg / day, 25-1000 mg / day, 50-1000 mg / day, 75-1000 mg / day, 100-1000 mg / day, 200-1000 mg / day, 25-800 mg / day, 50-800 mg / day, 100-800 mg / day, 200-800 mg / day, 25-400 mg / day, 50-400 mg / day, 100-400 mg / day, 200-400 mg / day. In some embodiments, the daily dose includes, but is not limited to, 1 mg / day, 5 mg / day, 10 mg / day, 20 mg / day, 25 mg / day, 50 mg / day, 75 mg / day, 100 mg / day, 125 mg / day, 150 mg / day, 200 mg / day, 400 mg / day, 600 mg / day, 800 mg / day, 1000 mg / day, 1200 mg / day, 1400 mg / day, 1440 mg / day.

[0038] The present invention relates to a kit, which may include a crystalline form in a single-dose or multi-dose form, the kit contains the inventive compound or crystalline form, and the amount of the inventive compound or crystalline form is the same as its amount in the above pharmaceutical composition.

[0039] In the present invention, the amount of the inventive compound or crystalline form is converted in the form of the free base in each case.

[0040] "Formulation specification" refers to the weight of the active ingredient contained in each vial, tablet or other unit preparation.

[0041] The crystalline form described in the present invention is present in an amount of about 5% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 10% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 15% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 20% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 25% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 30% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 35% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 40% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 45% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 50% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 55% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 60% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 65% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 70% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 75% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 80% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 85% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 90% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 95% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 98% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, it is present in an amount of about 99% to about 100% by weight of the active pharmaceutical ingredient; in certain embodiments, substantially all of the active pharmaceutical ingredient is a substantially pure compound or crystal.

[0042] The crystalline form of the present invention can be prepared by the following preparation methods:

[0043] 1. Volatilization experiment: Add the compound of formula I to a selected single solvent or binary solvent to form a clear sample solution, and volatilize it open to the air until the solvent is dry at different temperatures.

[0044] 2. Suspension method: Add the compound of formula I to a selected single solvent or binary solvent until a suspension is formed. After suspending and stirring for a certain period of time (such as 1 h to 3 days, or 2 h to 24 h, or 2 h to 12 h, or 3 to 5 h) at room temperature to 50 °C, centrifuge and separate the suspension, and dry it to obtain the product.

[0045] 3. Anti-solvent crystallization method: Dissolve the compound of formula I in a good solvent, take a certain amount of the solution and drop it into a poor solvent or drop the poor solvent into the solution, stir to precipitate a solid, separate and dry to obtain the product.

[0046] 4. Cooling method: Dissolve a certain amount of the sample in the corresponding solvent at a high temperature, transfer the solution to room temperature for cooling, let it stand or stir to crystallize, separate and dry to obtain the product.

[0047] 5. Thermal method experiment: Take a certain amount of the sample and place it on a glass slide on a hot stage, heat it to the target temperature at a certain rate (such as 5 - 20 °C / min, or 10 - 15 °C / min), and keep it at a constant temperature for a certain period of time (such as 0.5 - 5 min, or 1 - 3 min, or 1 - 2 min), then naturally cool it to room temperature to obtain a solid.

[0048] 6. Vapor diffusion experiment: At room temperature, add an appropriate amount of a good solvent to the compound of formula I dropwise to completely dissolve the sample or prepare a saturated solution of the good solvent; take a certain amount of the solution respectively, place the clear solution in the atmosphere of a poor solvent and let it stand at room temperature until a solid precipitates, then separate to obtain the product. Or directly place the solid of the compound of formula I in the atmosphere of the solvent and let it stand at room temperature for 1 - 7 days to obtain the product.

[0049] The good solvent and the poor solvent described in the present invention are relative. In a pair of solvents, the one with higher solubility is the good solvent, and the one with lower solubility is the poor solvent. In some embodiments, the good solvent is selected from the one with higher solubility among ethylene glycol methyl ether, ethylene glycol dimethyl ether, dioxane, DMF, DMSO, methanol, ethanol, n-propanol, butyl formate, 4-methyl-2-pentanone, tetrahydrofuran, isopropanol, ethyl acetate, n-heptane, ether, water, acetonitrile, toluene, chloroform, acetone, butyl formate, MTBE, cyclohexane, and the poor solvent is selected from the one with lower solubility among the above solvents. In some embodiments, the good solvent is selected from ethylene glycol methyl ether, ethylene glycol dimethyl ether, dioxane, DMF, DMSO, methanol, ethanol, n-propanol, butyl formate, 4-methyl-2-pentanone, tetrahydrofuran or their mixed solvents. In some embodiments, the poor solvent is selected from isopropanol, ethyl acetate, n-heptane, ether, water, acetonitrile, toluene, chloroform, acetone, butyl formate, MTBE, cyclohexane or their mixed solvents.

[0050] In some embodiments, the solvent for the evaporation method is water and acetone;

[0051] In some embodiments, the solvents used in the antisolvent crystallization method are dichloromethane and n-heptane; in some embodiments, the solvents used in the antisolvent crystallization method are dichloromethane and isopropyl ether; in some embodiments, the solvents used in the antisolvent crystallization method are ethyl acetate and n-heptane; in some embodiments, the solvents used in the antisolvent crystallization method are 4-methyl-2-pentanone and n-heptane; in some embodiments, the solvents used in the antisolvent crystallization method are dioxane and n-heptane;

[0052] In some embodiments, the solvents used in the cooling crystallization method are ethylene glycol dimethyl ether and cyclohexane; in some embodiments, the solvents used in the cooling crystallization method are toluene and cyclohexane; in some embodiments, the solvents used in the cooling crystallization method are isopropyl acetate and cyclohexane; in some embodiments, the solvents used in the cooling crystallization method are dioxane and cyclohexane; in some embodiments, the solvents used in the cooling crystallization method are dioxane and water; in some embodiments, the solvents used in the cooling crystallization method are DMSO and water.

[0053] In some embodiments, the gas phase diffusion method uses gas phase diffusion in ethyl acetate.

[0054] The good solvent and the poor solvent described in the present invention are relative. Among a pair of solvents, the one with higher solubility is the good solvent, and the one with lower solubility is the poor solvent.

[0055] For the solvents used in the above preparation method, when not specified, a single solvent can be used, or a combination of two or more solvents can be used.

[0056] The X-ray powder diffraction or DSC pattern, TGA pattern disclosed in the present invention, and those substantially the same as them also belong to the scope of the present invention.

[0057] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0058] “IC 50 ” refers to the half inhibitory concentration, which refers to the concentration when the maximum inhibitory effect reaches half.

[0059] As used in the present invention, “the crystal of the present invention”, “the crystal form of the present invention”, “the crystal form substance of the present invention” and the like can be used interchangeably.

[0060] “Room temperature” as described in the present invention generally refers to 4 - 30 °C, preferably 20 ± 5 °C.

[0061] The crystal form structure of the present invention can be analyzed using various analytical techniques known to those of ordinary skill in the art, including but not limited to, X-ray powder diffraction (XRD), differential scanning calorimetry (DSC), and / or thermogravimetric analysis (TGA), also known as thermogravimetry (TG).

[0062] As used herein, the term "2θ or 2θ angle" refers to the peak position in degrees (°) set in an X-ray diffraction experiment and is typically the horizontal axis unit in a diffraction pattern. If a reflection is diffracted when the incident beam makes an angle θ with a certain lattice plane, the experimental setup records the reflected beam at an angle of 2θ. It should be understood that the specific 2θ values mentioned herein for a particular crystal form are intended to represent the 2θ values (in degrees) measured using the X-ray diffraction experimental conditions described herein, and the error range of 2θ can be ±0.3, ±0.2, or ±0.1.

[0063] It is understood that the numerical values described and protected in the present invention are approximate values. Variations within the numerical values may be attributed to equipment calibration, equipment errors, crystal purity, crystal size, sample size, and other factors.

[0064] It is understood that the crystal forms of the present invention are not limited to the characteristic patterns exactly the same as those described in the accompanying drawings disclosed in the present invention. For example, for XRD, DSC, TGA, DVS, any crystal form having a characteristic pattern that is substantially the same or essentially the same as those described in the accompanying drawings falls within the scope of the present invention.

[0065] As is well known in the field of differential scanning calorimetry (DSC), the melting peak height of a DSC curve depends on many factors related to sample preparation and instrument geometry, while the peak position is relatively insensitive to experimental details. Therefore, in some embodiments, the crystalline compounds of the present invention are characterized by a DSC graph having a characteristic peak position, having substantially the same properties as the DSC graph provided in the accompanying drawings of the present invention, with a measurement error tolerance within ±5°C, and generally within ±3°C.

[0066] "Carrier" refers to a system that does not cause obvious irritation to the organism, does not eliminate the biological activity and characteristics of the administered compound, can change the way the drug enters the human body and its distribution in the body, control the release rate of the drug, and deliver the drug to the target organ. Non-limiting examples include microcapsules and microspheres, nanoparticles, liposomes, etc.

[0067] "Excipient" means: a substance that is not itself a therapeutic agent and is used as a diluent, adjuvant, binder, and / or vehicle, added to a pharmaceutical composition to improve its handling or storage properties or to permit or facilitate the formation of a unit dosage form of a compound or pharmaceutical composition for administration. As is known to those skilled in the art, pharmaceutical excipients can provide various functions and can be described as wetting agents, buffers, suspending agents, lubricants, emulsifiers, disintegrants, absorbents, preservatives, surfactants, coloring agents, flavoring agents, and sweetening agents. Examples of pharmaceutical excipients include, but are not limited to: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, cellulose acetate, hydroxypropyl methyl cellulose, hydroxypropyl cellulose, microcrystalline cellulose, and cross-linked carboxymethyl cellulose (e.g., sodium cross-linked carboxymethyl cellulose); (4) tragacanth powder; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) diols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) pH buffer solutions; (21) polyesters, polycarbonates, and / or polyanhydrides; and (22) other non-toxic and compatible substances used in pharmaceutical formulations. Description of the Drawings

[0068] Figure 1 X-ray powder diffraction pattern of the amorphous form of the compound of formula (I).

[0069] Figure 2 X-ray powder diffraction pattern of polymorph I of the compound of formula (I).

[0070] Figure 3 Differential scanning calorimetry curve pattern of polymorph I of the compound of formula (I).

[0071] Figure 4 Thermogravimetric analysis pattern of polymorph I of the compound of formula (I).

[0072] Figure 5 Isothermal adsorption curve of polymorph I of the compound of formula (I).

[0073] Figure 6 X-ray powder diffraction pattern of polymorph II of the compound of formula (I).

[0074] Figure 7 Differential scanning calorimetry curve atlas of polymorph II of the compound shown in formula (I).

[0075] Figure 8 Thermogravimetric analysis atlas of polymorph II of the compound shown in formula (I).

[0076] Figure 9 Isothermal adsorption curve of polymorph II of the compound shown in formula (I). Detailed implementation manners

[0077] The structure of the compound is determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). The NMR shift (δ) is given in units of 10 -6 (ppm). The NMR measurement is performed using (Bruker Avance III 400 and Bruker Avance 300) nuclear magnetic resonance spectrometers, and the solvents for measurement are deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), and the internal standard is tetramethylsilane (TMS).

[0078] The MS measurement is performed using (Agilent 6120B (ESI) and Agilent 6120B (APCI)).

[0079] The HPLC measurement uses an LC-20AT (Shimadzu) high-performance liquid chromatograph (Shim-pack GIST C18, 4.6×250 mm (HSS), 5 μm).

[0080] Crystal form test examples

[0081] 1. Instrument information and detection method parameter table (see Table 1 below)

[0082] Table 1 Instrument information and detection method parameter table

[0083]

[0084]

[0085]

[0086]

[0087] The known starting materials of the present invention can be adopted or synthesized according to methods known in the art, or can be purchased from companies such as Titan Technology, Energy Chemical, Shanghai Dermer, Chengdu Kelong Chemical Industry, Shaoyuan Chemical Technology, and J&K Scientific.

[0088] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0089] Unless otherwise specified in the examples, the room temperature is 20°C to 30°C.

[0090] The known starting materials of the present invention can be used or synthesized according to methods known in the art, or can be purchased from companies such as Titan Technology, Energy Chemical, Shanghai Dermer, Chengdu Kelong Chemical Industry, Shaoyuan Chemical Technology, and J&K Scientific.

[0091] The following specifically illustrates the implementation process and beneficial effects of the present invention through specific examples, aiming to help readers better understand the essence and characteristics of the present invention, and shall not be used as a limitation on the scope of implementation of this case.

[0092] Example 1: Preparation of the compound of formula (I)

[0093]

[0094] Synthesis of the compound of formula I:

[0095] First step: 6-bromo-2-(trifluoromethyl)imidazo[1,2-a]pyrazine (1A)

[0096]

[0097] Dissolve 3-bromo-1,1,1-trifluoroacetone (109.74 g, 574.71 mmol) and 2-amino-5-bromopyrazine (20 g, 114.94 mmol) in 1,4-dioxane (100 mL), and stir at 100°C for 1 h. Monitor the reaction by LCMS, filter and collect the filter cake. The filter cake is dispersed in 1,4-dioxane (300 mL) and stirred at 100°C overnight. After the reaction is completed, directly evaporate to dryness and purify by flash column chromatography (PE / EA = 0 - 20%) to obtain compound 1A (24 g, 80%).

[0098] MS m / z = 266.0 [M+1] +

[0099] Second step: tert-butyl 4-(2-(trifluoromethyl)imidazo[1,2-a]pyrazin-6-yl)piperazine-1-carboxylate (1B)

[0100]

[0101] Compound 1B (5 g, 18.8 mmol), tert-butyl piperazine-1-carboxylate (4.21 g, 22.56 mmol), tris(dibenzylideneacetone)dipalladium(0) (2.58 g, 2.82 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (3.27 g, 5.64 mmol), and sodium tert-butoxide (2.18 g, 22.56 mmol) were added to a reaction flask and stirred at 100 °C for 1 h under nitrogen protection. After the reaction was completed, the reaction solution was extracted three times with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography (EA:PE = 20%) to obtain Compound 1B (800 mg, 11%).

[0102] LCMS m / z = 372.1 [M+1] +

[0103] Step 3: 6-(Piperazin-1-yl)-2-(trifluoromethyl)imidazo[1,2-a]pyrazine (1C)

[0104]

[0105] Compound 1C (330 mg, 0.89 mmol) and trifluoroacetic acid (3 mL) were added to a reaction flask and stirred at 25 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain crude Compound 1C (240 mg, 100%), which was directly used in the next reaction step.

[0106] Step 4: 8-Fluoro-3-methyl-7-((4-(2-(trifluoromethyl)imidazo[1,2-a]pyrazin-6-yl)piperazin-1-yl)methyl)quinoxalin-2(1H)-one (Formula I)

[0107] 8-fluoro-3-methyl-7-((4-(2-(trifluoromethyl)imidazo[1,2-a]pyrazin-6-yl)piperazin-1-yl)methyl)quinoxalin-2(1H)-one

[0108]

[0109] 1D (synthesized according to Patent US2022 / 0009901 A1) (1.2 g, 4.43 mmol), 1C (1 g, 3.69 mmol), N,N - diisopropylethylamine (1.43 g, 11.07 mmol), potassium iodide (61.42 mg, 0.37 mmol), and acetonitrile (3 mL) were added to a reaction flask and stirred at 60 °C for 1 h. The reaction was monitored by LCMS. After the reaction was completed, the reaction system was directly sent for preparation. The preparative HPLC separation method: 1. Instrument: waters 2767 preparative liquid phase; Chromatographic column: SunFire@Prep C18 (19 mm × 250 mm) 2. The sample was filtered through a 0.45 μm filter head to prepare a sample solution. 3. Preparative chromatographic conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile; Mobile phase B: water (containing 0.1% ammonium acetate) b. Gradient elution, the content of mobile phase A ranged from 10% - 55% c. Flow rate 12 mL / min. The title compound of formula I (1 g, 58%) was obtained at a retention time of 7.0 min.

[0110] LCMS m / z = 462.5 [M + 1] +

[0111] 1 H NMR (400 MHz, DMSO - d6) δ = 8.98 (s, 1H), 8.45 (s, 1H), 7.91 (s, 1H), 7.52 (d, 1H), 7.31 (t, 1H), 3.70 (s, 2H), 3.32 (s, 3H), 3.28 (s, 2H), 2.62–2.55 (m, 4H), 2.42 (s, 3H).

[0112] Preparation of the amorphous form of the compound of formula (I) in Example 2

[0113] Take 50 mg of the compound of formula (I) obtained in Example 1, add it to a mixed solvent of 20 mL of acetonitrile and water. After dissolving clearly, it was freeze - dried. The compound of formula (I) was characterized by XRD as amorphous, as Figure 1 。

[0114] Preparation of polymorphic form I of the compound of formula (I) in Example 3

[0115] Take 150 mg of the compound of formula (I) obtained in Example 1, add it to a mixed solvent of 4 mL of acetonitrile and water, dissolve it at 50 °C, filter it. The obtained filtrate was cooled and crystallized at room temperature, stirred at room temperature overnight, centrifuged. The obtained solid was vacuum - dried at 50 °C overnight to obtain polymorphic form I of the compound of formula (I). The polymorphic form I of the compound of formula (I) was characterized by XRD, DSC, TGA, and DVS, successively as Figure 2 - 5 。

[0116] 11H NMR (400 MHz, DMSO-d6) δ = 8.98 (s, 1H), 8.45 (s, 1H), 7.91 (s, 1H), 7.52 (d, 1H), 7.31 (t, 1H), 3.70 (s, 2H), 3.32 (s, 3H), 3.28 (s, 2H), 2.62–2.55 (m, 4H), 2.42 (s, 3H).

[0117] The X-ray powder diffraction pattern (XRD) of polymorph I of the compound of formula (I) is as Figure 2 shown. The specific peaks are shown in Table 2.

[0118] Table 2 Specific XRD Peaks of Polymorph I

[0119]

[0120]

[0121] Preparation of Polymorph II of the Compound of Formula (I) in Example 4

[0122] Take 150 mg of the compound of formula (I) obtained in Example 1, add dichloromethane (3 mL) and methanol (1 mL), dissolve at 50 °C, add methyl tert-butyl ether (2.0 mL), precipitate a solid, stir at room temperature overnight, centrifuge, and vacuum dry the obtained solid at 50 °C overnight to obtain polymorph II of the compound of formula (I). The polymorph II of the compound of formula (I) was characterized by XRD, DSC, TGA, and DVS, successively as Figure 6 - 9 .

[0123] 1 1H NMR (400 MHz, DMSO-d6) δ = 8.98 (s, 1H), 8.45 (s, 1H), 7.91 (s, 1H), 7.52 (d, 1H), 7.31 (t, 1H), 3.70 (s, 2H), 3.32 (s, 3H), 3.28 (s, 2H), 2.62–2.55 (m, 4H), 2.42 (s, 3H).

[0124] The X-ray powder diffraction pattern (XRD) of polymorph II of the compound of formula (I) is as Figure 6 shown. The specific peaks are shown in Table 3.

[0125] Table 3 Specific XRD Peaks of Polymorph II

[0126]

[0127]

[0128] Example 5 Study on the Stability of Related Polymorphs

[0129] 1. The stability of Form I of the compound of formula (I) was

[0130] determined under the conditions shown in Table 4, and the experimental results are shown in Table 5.

[0131] Table 4 Solid-state stability experiment of Form I

[0132]

[0133] Table 5. Detection results of the solid-state stability experiment of Form I

[0134]

[0135] Conclusion: Form I of the compound of formula (I) has good solid-state stability.

[0136] 2. The stability of Form II of the compound of formula (I) was

[0137] tested under the conditions shown in Table 6, and the experimental results are shown in Table 7.

[0138] Table 6 Solid-state stability experiment of Form II

[0139]

[0140] Table 7. Detection results of the solid-state stability experiment of Form II

[0141]

[0142]

[0143] Conclusion: The solid-state stability of Form II of the compound of formula (I) is good.

[0144] Example 6. Polymorphic transformation study of the compound of formula (I)

[0145] 10 mg samples of Form I and 10 mg samples of Form II were taken respectively, mixed evenly, and samples were taken for XRPD characterization; the mixed samples were added to 0.5 mL of a mixed solvent of dimethyl sulfoxide and water (1:1) to form a suspension, stirred at room temperature for 5 days, centrifuged, and vacuum dried at 40 °C overnight, and samples were taken for XRPD characterization.

[0146] Table 8. Table of experimental results of polymorphic transformation of Form I and Form II

[0147] Condition Temperature Dimethyl sulfoxide / water (1:1) Result Room temperature Crystal form 1

[0148] Conclusion: Under room temperature conditions, the thermodynamic stability of Form I is better than that of Form II.

[0149] Example 7 PARP-1 enzyme activity test experiment

[0150] PARP-1 chemiluminescence detection kit was purchased from BPS Bioscience. The histone solution in the kit was diluted 5 times with 1XPBS, 25 μL of histone dilution was taken to the microplate, and incubated at 4°C overnight. After the incubation, the plate was washed 3 times with PBST (0.05% Tween-20), 100 μL of blocking solution was taken to the microplate, and incubated at 25°C for 90 minutes; after the incubation, the plate was washed 3 times with PBST. 2.5 μL of compounds of different concentrations diluted in the test buffer and 12.5 μL of substrate mixed solution (1.25 μL 10XPARP test buffer; 1.25 μL 10X PARP test mixture; 2.5 μL Activated DNA, 7.5 μL double distilled water) were taken to the microplate. The PARP-1 enzyme was diluted to 2 ng / μL, 10 μL was taken to the microplate, and the reaction system was incubated at 25°C for 60 minutes;

[0151] After incubation, wash the plate three times with PBST. Dilute Streptavidin-HRP 50-fold with blocking buffer, then transfer 25 μL to the microplate and incubate at 25°C for 30 minutes. After incubation, wash the plate three times with PBST. Mix ELISA ECL substrate A and substrate B at a 1:1 (v / v) ratio, transfer 50 μL to the microplate, and read the chemiluminescence value.

[0152] The inhibition rate was calculated according to formula 1, where RLUsample is the compound well reading, RLUmax is the solvent control well reading, and RLUmin is the PARP-1 enzyme-free control well reading. GraphPad Prism software was used to perform curve fitting using four parameters (log (inhibitor) vs. response--Variable slope) and calculate the IC 50 value.

[0153] Inhibition%=(1-(RLUsample-RLUmin) / (RLUmax-RLUmin))×100% (Formula 1)

[0154] Experimental results: The IC of compound (I) against PARP-1 in vitro 50 The value is expressed as A, B, C, D, with A representing 0 <IC 50 ≤5nM, B means 5nM <IC 50 ≤10nM, C represents 10nM <IC 50 ≤50nM, D means 50nM <IC 50 ≤100nM.

[0155] Table 9 PARP-1 enzyme activity

[0156] Compound <![CDATA[IC 50 (nM)]]> Compound of formula (I) A

[0157] Conclusion: The compound of formula (I) of the present invention has a significant inhibitory effect on PARP-1 enzyme activity in vitro.

[0158] Example 8 Mouse Pharmacokinetic Test

[0159] Test animals: Male Balb / c mice, 20 - 25 g, 12 mice / compound. Purchased from Chengdu Dashuo Laboratory Animal Co., Ltd.

[0160] Test design: On the day of the test, Balb / c mice were randomly grouped according to body weight. They were fasted for 12 - 14 h without water deprivation 1 day before dosing and fed 4 h after dosing.

[0161] Table 10. Dosing Information

[0162]

[0163] Note: Intravenous administration vehicle: 10% DMA + 10% Solutol + 80% Saline; Gavage administration vehicle: 5% DMSO + 30% PEG400 + 65% (20% SBE-CD), Solutol is polyethylene glycol-15-hydroxystearate; Saline is physiological saline

[0164] Blood (0.06 mL) was collected from the orbital cavity under isoflurane anesthesia before and after dosing and placed in an EDTAK2 centrifuge tube. It was centrifuged at 5000 rpm at 4°C for 10 min to collect plasma. The blood sampling time points for both the intravenous group and the gavage group were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 24 h. Brains were collected at 30 min, 2, 24 h after administration of Compounds 12, 15, 18 respectively. Brains of Compounds 2 and 5 were collected at 24 h. The surface residual blood of the brains was rinsed with cold physiological saline, blotted dry, and homogenized. Before analysis and detection, all samples were stored at -80°C, and LC-MS / MS was used for quantitative analysis of the samples.

[0165] Table 11. Pharmacokinetic Parameters of Test Compounds in Mouse Plasma

[0166]

[0167] Conclusion: The compound of formula (I) has excellent pharmacokinetic characteristics in mouse plasma.

Claims

1. A compound of formula (I):

2. The compound of formula (I) according to claim 1, which is a crystalline form.

3. The crystalline form according to claim 2, wherein The crystalline form is crystalline form I. Using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 9.17° ± 0.2°, 10.37° ± 0.2°, 13.84° ± 0.2°, 15.80° ± 0.2°, 16.24° ± 0.2°, 17.05° ± 0.2°, and 17.92° ± 0.2°; preferably, it has characteristic diffraction peaks at the following 2θ positions: 9.17° ± 0.2°, 10.37° ± 0.2°, 13.84° ± 0.2°, 16.24° ± 0.2°, 17.05° ± 0.2°, 17.92° ± 0.2°, 18.45° ± 0.2°, 20.41° ± 0.2°, 21.47° ± 0.2°, 22.18° ± 0.2°, 22.47° ± 0.2°, 24.34° ± 0.2°, 24.95° ± 0.2°, 25.24° ± 0.2°, and 25.97° ± 0.2°.

4. The crystalline form according to claim 3. Using Cu-Kα radiation, its X-ray powder diffraction pattern is substantially as shown in Figure 2; preferably, its differential scanning calorimetry curve, thermogravimetric analysis curve, and isothermal adsorption curve are substantially as shown in Figures 3, 4, and 5 respectively.

5. The crystalline form according to claim 2, wherein, The crystalline form is crystalline form II. Using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 7.49° ± 0.2°, 11.23° ± 0.2°, 14.76° ± 0.2°, 17.63° ± 0.2°, 18.77° ± 0.2°, 21.43° ± 0.2°, 21.71° ± 0.2°, and 24.32° ± 0.2°; preferably, it has characteristic diffraction peaks at the following 2θ positions: 7.49° ± 0.2°, 11.23° ± 0.2°, 11.83° ± 0.2°, 14.76° ± 0.2°, 17.63° ± 0.2°, 18.77° ± 0.2°, 21.43° ± 0.2°, 21.71° ± 0.2°, 22.39° ± 0.2°, 24.32° ± 0.2°, 25.34° ± 0.2°, and 25.72° ± 0.2°.

6. The crystalline form according to claim 5. Using Cu-Kα radiation, its X-ray powder diffraction pattern is substantially as shown in Figure 6; preferably, its differential scanning calorimetry curve, thermogravimetric analysis curve, and isothermal adsorption curve are substantially as shown in Figures 7, 8, and 9 respectively.

7. A pharmaceutical composition comprising a therapeutically effective amount of the compound or crystalline form according to any one of claims 1-6, and a pharmaceutically acceptable carrier and / or excipient, wherein the therapeutically effective amount is preferably 1-1440 mg.

8. Use of the compound or crystalline form according to any one of claims 1-6, or the pharmaceutical composition according to claim 7, in the preparation of a medicament for treating PARP-mediated diseases.

9. A method for treating PARP-mediated diseases, the method comprising administering to a subject a therapeutically effective amount of the compound or crystal form according to any one of claims 1-6, the therapeutically effective amount being preferably 1-1440 mg, and the disease being preferably breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer.

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