Crystalline form of 7-azaspiro[4,5]decane-6,10-dione compound and its preparation method

Abstract

In the present invention, crystalline forms of 7-azaspiro[4,5]decane-6,10-dione compounds and methods for their preparation are disclosed. In particular, methods for the preparation of the compound of formula (I) and its crystalline forms, as well as uses of said compound and its crystalline forms are disclosed. TIFF2025504040000023.tif3356(I)

Description

[Technical field]

[0001] This application is: CN202210103134.0 filed on January 27, 2022; CN202211017556.2 filed on August 23, 2022 claims priority to the

[0002] The present application discloses crystalline forms of 7-azaspiro[4,5]decane-6,10-dione compounds and methods for their preparation, and in particular discloses methods for the preparation and use of the compound of formula (I) and its crystalline forms: [Background technology]

[0003] Hypertrophic cardiomyopathy (HCM) is a cardiac disease characterized by hypertrophy of the myocardium, often eroding the interventricular septum, narrowing the ventricular cavity, impeding blood filling of the left ventricle, and reducing the diastolic compliance of the left ventricle. Hypertrophic cardiomyopathy is divided into obstructive and nonobstructive types, depending on the presence or absence of obstruction of the left ventricular outflow tract, which may be related to genetics. HCM cases worldwide account for approximately 1 / 500, and the clinical manifestations of HCM are diverse. It may be asymptomatic or may cause palpitations, exertional dyspnea, dull precordial pain, fatigue, syncope, and even sudden death. In the end stage, symptoms of heart failure are seen in the left side of the heart.

[0004] Currently, there are limited drugs for the treatment of HCM. The treatment of HCM is mainly to improve symptoms through beta blockers or calcium channel blockers, but it cannot target the cause, cannot slow the progression of myocardial hypertrophy, and cannot improve prognosis. The therapeutic effect is limited.

[0005] Myosin and actin are the basis of myocardial contraction. Myosin crossbridges periodically bind and dissociate with actin, driving the sliding of myofilaments and inducing myocardial contraction. Myosin has ATPase activity and provides the power of myocardial contraction through ATP hydrolysis. Myosin mutations lead to the extension of the binding time of myosin and actin, excessive contraction and impaired relaxation of the left ventricular myocardium, resulting in left ventricular hypertrophy and fibrosis, and inducing HCM. MYK-461 is an allosteric regulator of cardiac myosin, which slows down the rate of phosphate hydrolysis, shortens the binding time of myosin and actin, produces a negative inotropic effect, and alleviates pathological changes, such as myocardial hypertrophy caused by excessive contraction of the left ventricular myocardium. However, the drug is slowly excreted from the body and stays in the body for too long, making it difficult to adjust the dose quickly. Therefore, the development of myosin inhibitors with better activity and more ideal pharmacokinetic properties is of great clinical value and significance.

[0006] Furthermore, abnormalities in myocardial sarcomeres have been identified as a driving factor in various cardiac diseases and conditions, such as diastolic heart failure with preserved ejection fraction, ischemic heart disease, angina pectoris, and restrictive cardiomyopathy, etc. Myosin ATPase inhibitors may also have potential therapeutic effects in alleviating the pathological processes of the above diseases by inhibiting myocardial contraction. Summary of the Invention

[0007] The present disclosure relates to a compound of formula (I), characterized by an X-ray powder diffraction pattern (XRPD) having characteristic diffraction peaks at angles 2θ of 18.283±0.200°, 19.662±0.200°, and 22.420±0.200°; [ka] (I) The present invention provides crystalline form A of the compound of the formula:

[0008] In some embodiments of the present disclosure, the crystalline form A has an X-ray powder diffraction pattern with characteristic diffraction peaks at angles 2θ of 11.143±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 26.259±0.200°, and 28.056±0.200°.

[0009] In some embodiments of the present disclosure, the crystalline form A has an X-ray powder diffraction pattern with characteristic diffraction peaks at angles 2θ of 11.143±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 26.259±0.200°, 27.261±0.200°, 28.056±0.200°, and 30.256±0.200°.

[0010] In some embodiments of the present disclosure, crystalline form A is disclosed, characterized by an X-ray powder diffraction pattern comprising at least 5, 6, 7, or 8 characteristic diffraction peaks at angles 2θ selected from the following angles: 11.143±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 26.259±0.200°, 27.261±0.200°, 28.056±0.200°, and 30.256±0.200°.

[0011] In some embodiments of the present disclosure, the crystalline form A has an X-ray powder diffraction pattern with characteristic diffraction peaks at angles 2θ of 11.143±0.200°, 16.522±0.200°, 17.053±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 23.909±0.200°, 26.259±0.200°, 27.261±0.200°, 28.056±0.200°, 30.256±0.200°, and 33.761±0.200°.

[0012] In some embodiments of the present disclosure, the crystalline form A is characterized by an X-ray powder diffraction pattern comprising at least 5, 6, 7, 8, 9, 10, 11, or 12 characteristic diffraction peaks at angles 2θ selected from the following angles: 11.143±0.200°, 16.522±0.200°, 17.053±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 23.909±0.200°, 26.259±0.200°, 27.261±0.200°, 28.056±0.200°, 30.256±0.200°, and 33.761±0.200°.

[0013] In some embodiments of the present disclosure, the crystalline form A has a molecular weight of 11.143±0.200°, 13.260±0.200°, 16.522±0.200°, 17.053±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 23.568±0.200°, 23.909±0.200°, 24.04±0.200°, 25.04±0.200°, 26.04±0.200°, 27.04±0.200°, 28.04±0.200°, 29.04±0.200°, 30.04±0.200°, 31.04±0.200°, 32.04±0.200°, 33.04±0.200°, 34.04±0.200°, 35.04±0.200°, 36.04±0.200°, 37.04±0.200°, 38.04±0.200°, 39.04±0.200°, 40.04±0.200°, 41.04±0.200°, 42.04±0.200°, 43.04±0.200°, 44.04±0.200°, 45.04±0.200°, 46.04±0.200°, 47.04±0.200°, 48.04±0.200°, 49.04±0 It has an X-ray powder diffraction pattern with characteristic diffraction peaks at angles 2θ of 0.200°, 26.259±0.200°, 27.261±0.200°, 28.056±0.200°, 29.008±0.200°, 30.256±0.200°, 33.761±0.200°, and 35.404±0.200°.

[0014] In some embodiments of the present disclosure, the crystalline form A has the following molecular weights: 11.143±0.200°, 13.260±0.200°, 14.860±0.200°, 16.163±0.200°, 16.522±0.200°, 17.053±0.200°, 17.537±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 23.568±0.200°, 23.909±0.200°, 26.259±0.200°, 26.726±0.200°, 27.261±0.200°, It has an X-ray powder diffraction pattern with characteristic diffraction peaks at angles 2θ of 28.056±0.200°, 29.008±0.200°, 30.256±0.200°, 31.219±0.200°, 31.646±0.200°, 32.037±0.200°, 32.438±0.200°, 32.807±0.200°, 33.761±0.200°, 34.534±0.200°, 35.404±0.200°, 36.856±0.200°, 37.813±0.200°, and 39.456±0.200°.

[0015] In some embodiments of the present disclosure, the crystalline form A has the following molecular weights: 11.143±0.100°, 13.260±0.100°, 14.860±0.100°, 16.163±0.100°, 16.522±0.100°, 17.053±0.100°, 17.537±0.100°, 18.283±0.100°, 19.662±0.100°, 22.420±0.100°, 23.568±0.100°, 23.909±0.100°, 26.259±0.100°, 26.726±0.100°, 27.261±0.100°, It has an X-ray powder diffraction pattern with characteristic diffraction peaks at angles 2θ of 28.056±0.100°, 29.008±0.100°, 30.256±0.100°, 31.219±0.100°, 31.646±0.100°, 32.037±0.100°, 32.438±0.100°, 32.807±0.100°, 33.761±0.100°, 34.534±0.100°, 35.404±0.100°, 36.856±0.100°, 37.813±0.100°, and 39.456±0.100°.

[0016] In some embodiments of the present disclosure, the crystalline form A has the following molecular weights: 11.143°, 13.260°, 14.860°, 16.163°, 16.522°, 17.053°, 17.537°, 18.283°, 19.662°, 22.420°, 23.568°, 23.909°, 26.259°, 26.726°, 27.261°, 28. It has a powder X-ray diffraction pattern with characteristic diffraction peaks at angles 2θ of 056°, 29.008°, 30.256°, 31.219°, 31.646°, 32.037°, 32.438°, 32.807°, 33.761°, 34.534°, 35.404°, 36.856°, 37.813°, and 39.456°.

[0017] In some embodiments of the present disclosure, the crystalline form A has an average crystal orientation of 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, and / or 11.143±0.200°, and / or 13.260±0.200°, and / or 14.860±0.200°, and / or 16.163±0.200°, and / or 16.5 22±0.200°, and / or 17.053±0.200°, and / or 17.537±0.200°, and / or 23.568±0.200°, and / or 23.909±0.200°, and / or 26.259±0.200°, and / or 26.726±0.200°, and / or 27.261±0.200°, and / or is 28.056 ± 0.200°, and / or 29.008 ± 0.200°, and / or 30.256 ± 0.200°, and / or 31.219 ± 0.200°, and / or 31.646 ± 0.200°, and / or 32.037 ± 0.200°, and / or 32.438 ± 0.200°, and / or 32.807 ± 0.200°, and / or has an X-ray powder diffraction pattern having characteristic diffraction peaks at angles 2θ of 33.761±0.200°, and / or 34.534±0.200°, and / or 35.404±0.200°, and / or 36.856±0.200°, and / or 37.813±0.200°, and / or 39.456±0.200°.

[0018] In some embodiments of the present disclosure, the crystalline form A has an average crystal orientation of 18.283±0.100°, 19.662±0.100°, 22.420±0.100°, and / or 11.143±0.100°, and / or 13.260±0.100°, and / or 14.860±0.100°, and / or 16.163±0.100°, and / or 16.5 22±0.100°, and / or 17.053±0.100°, and / or 17.537±0.100°, and / or 23.568±0.100°, and / or 23.909±0.100°, and / or 26.259±0.100°, and / or 26.726±0.100°, and / or 27.261±0.100°, and / or is 28.056 ± 0.100°, and / or 29.008 ± 0.100°, and / or 30.256 ± 0.100°, and / or 31.219 ± 0.100°, and / or 31.646 ± 0.100°, and / or 32.037 ± 0.100°, and / or 32.438 ± 0.100°, and / or 32.807 ± 0.100°, and / or has an X-ray powder diffraction pattern having characteristic diffraction peaks at angles 2θ of 33.761±0.100°, and / or 34.534±0.100°, and / or 35.404±0.100°, and / or 36.856±0.100°, and / or 37.813±0.100°, and / or 39.456±0.100°.

[0019] In some embodiments of the present disclosure, the XRPD pattern of crystalline form A is substantially as shown in FIG.

[0020] In some embodiments of the present disclosure, the resolution data of the XRPD pattern for crystalline form A is shown in Table 1. Table 1. XRPD pattern resolution data for crystalline form A of compound of formula (I) [Table 1] [Table 2]

[0021] In some embodiments of the present disclosure, the crystalline form A is disclosed having a differential scanning calorimetry curve with endothermic peak values ​​at 249.53±8°C and 306.12±8°C.

[0022] In some embodiments of the present disclosure, the crystalline form A is disclosed having a differential scanning calorimetry curve with endothermic peak values ​​at 249.53±3°C and 306.12±3°C.

[0023] In some embodiments of the present disclosure, crystalline form A is disclosed, having a DSC curve substantially as shown in FIG.

[0024] In some embodiments of the present disclosure, the crystalline form A is disclosed having a thermogravimetric analysis curve with a maximum weight loss of 0.075% at 200±3°C and a maximum weight loss of 0.164% at 250±3°C.

[0025] In some embodiments of the present disclosure, crystalline form A is disclosed, having a TGA curve substantially as shown in FIG. [ka] (I)

[0026] In some embodiments of the present disclosure, the crystalline form B has an X-ray powder diffraction pattern with characteristic diffraction peaks at angles 2θ of 12.861±0.200°, 14.135±0.200°, 17.788±0.200°, 18.099±0.200°, 21.659±0.200°, 21.912±0.200°, 22.410±0.200°, and 25.218±0.200°.

[0027] In some embodiments of the present disclosure, crystalline form B is disclosed, characterized by an X-ray powder diffraction pattern comprising at least 5, 6, 7, or 8 characteristic diffraction peaks at angles 2θ selected from the following angles: 12.861±0.200°, 14.135±0.200°, 17.788±0.200°, 18.099±0.200°, 21.659±0.200°, 21.912±0.200°, 22.410±0.200°, and 25.218±0.200°.

[0028] In some embodiments of the present disclosure, the crystalline form B has an X-ray powder diffraction pattern with characteristic diffraction peaks at angles 2θ of 8.262±0.200°, 12.861±0.200°, 14.135±0.200°, 16.303±0.200°, 17.788±0.200°, 18.099±0.200°, 19.053±0.200°, 21.659±0.200°, 21.912±0.200°, 22.410±0.200°, 25.218±0.200°, 27.230±0.200°, 27.526±0.200°, and 28.132±0.200°.

[0029] In some embodiments of the present disclosure, the following angles are used: 8.262±0.200°, 12.861±0.200°, 14.135±0.200°, 16.303±0.200°, 17.788±0.200°, 18.099±0.200°, 19.053±0.200°, 21.659±0.200°, 21.912±0.200°, 22.410±0.200°, 23.01±0.200°, 24.01±0.200°, 25.01±0.200°, 26.01±0.200°, 27.01±0.200°, 28.01±0.200°, 29.01±0.200°, 30.01±0.200°, 31.01±0.200°, 32.01±0.200°, 33.01±0.200°, 34.01±0.200°, 35.01±0.200°, 36.01±0.200°, 37.01±0.200°, 38.01±0.200°, 39.01±0.200°, 40.01±0.200°, 41.01±0.200°, 42.01±0.200°, 43.01±0.200°, 44.01±0.200°, 45.01±0.200°, 46.01±0.200°, 47.01±0.200 The crystalline form B is disclosed as being characterized by a powder X-ray diffraction pattern comprising at least 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 characteristic diffraction peaks representative of angles 2θ selected from 200°, 25.218±0.200°, 27.230±0.200°, 27.526±0.200°, and 28.132±0.200°.

[0030] In some embodiments of the present disclosure, the crystalline form B has a molecular weight of 7.240±0.200°, 8.262±0.200°, 9.189±0.200°, 10.841±0.200°, 12.861±0.200°, 13.269±0.200°, 14.135±0.200°, 14.651±0.200°, 15.907±0.200°, 16.303±0.200°, 16.770±0.200°, 17.788±0.200°, 18.099±0.200°, 19.053±0.200°, 21.659±0.200°, 22.770±0.200°, 23.770±0.200°, 24.770±0.200°, 25.770±0.200°, 26.770±0.200°, 27.770±0.200°, 28.770±0.200°, 29.770±0.200°, 30.770±0.200°, 31.770±0.200°, 32.770±0.200°, 33.770±0.200°, 34.770±0.200°, 35.770±0.200°, 36.770±0.200°, 37.770±0.200°, 38.770±0.200°, 39.770±0.200°, and 33.645±0.200°.

[0031] In some embodiments of the present disclosure, the crystalline form B has a molecular weight of 7.240±0.100°, 8.262±0.100°, 9.189±0.100°, 10.841±0.100°, 12.861±0.100°, 13.269±0.100°, 14.135±0.100°, 14.651±0.100°, 15.907±0.100°, 16.303±0.100°, 16.770±0.100°, 17.788±0.100°, 18.099±0.100°, 19.053±0.100°, 21.659±0.100°, 22.777±0.100°, 23.777±0.100°, 24.777±0.100°, 25.777±0.100°, 26.777±0.100°, 27.777±0.100°, 28.777±0.100°, 29.777±0.100°, 30.777±0.100°, 31.777±0.100°, 32.777±0.100°, 33.777±0.100°, 34.777±0.100°, 35.777±0.100°, 36.777±0.100°, 37.777±0.100°, 38.777±0.100°, 39.777±0.100°, It has a powder X-ray diffraction pattern with characteristic diffraction peaks at angles 2θ of 0.100°, 21.912±0.100°, 22.410±0.100°, 22.829±0.100°, 24.408±0.100°, 25.218±0.100°, 26.009±0.100°, 27.230±0.100°, 27.526±0.100°, 28.132±0.100°, 29.630±0.100°, 32.150±0.100°, 32.961±0.100°, and 33.645±0.100°.

[0032] In some embodiments of the present disclosure, the crystalline form B has the following molecular weights: 7.240°, 8.262°, 9.189°, 10.841°, 12.861°, 13.269°, 14.135°, 14.651°, 15.907°, 16.303°, 16.770°, 17.788°, 18.099°, 19.053°, 21.659°, It has a powder X-ray diffraction pattern with characteristic diffraction peaks at angles 2θ of 21.912°, 22.410°, 22.829°, 24.408°, 25.218°, 26.009°, 27.230°, 27.526°, 28.132°, 29.630°, 32.150°, 32.961°, and 33.645°.

[0033] In some embodiments of the present disclosure, the crystalline form B has an average crystallinity of 17.788°±0.200°, 18.099±0.200°, 21.659±0.200°, and / or 21.912±0.200°, and / or 7.240±0.200°, and / or 8.262±0.200°, and / or 9.189±0.200°, and / or 10.841±0.200°, and / or 12.861±0.200°, and / or 13.269±0.200°, and / or 14.135±0.200°, and / or 14.651±0.200°, and / or 15.907±0.200°, and / or 16.303±0.200°, and / or 16.770±0.2 and / or 27.526±0.200°, and / or 28.132±0.200°, and / or 29.630±0.200°, and / or 32.150±0.200°, and / or 32.961±0.200°, and / or 33.645±0.200°.

[0034] In some embodiments of the present disclosure, the crystalline form B has an angle of 17.788°±0.100°, 18.099±0.100°, 21.659±0.100°, and / or 21.912±0.100°, and / or 7.240±0.100°, and / or 8.262±0.100°, and / or 9.189±0.100°, and / or 10.841±0.100°, and / or 12.861±0.100°, and / or 13.269±0.100°, and / or 14.135±0.100°, and / or 14.651±0.100°, and / or 15.907±0.100°, and / or 16.303±0.100°, and / or 16.770±0.1 and / or 27.526±0.100°, and / or 28.132±0.100°, and / or 29.630±0.100°, and / or 32.150±0.100°, and / or 32.961±0.100°, and / or 33.645±0.100°.

[0035] In some embodiments of the present disclosure, the resolution data of the XRPD pattern of crystalline form B is shown in Table 2. Table 2. XRPD pattern resolution data for crystalline form B of compound of formula (I) [Table 3] [Table 4]

[0036] In some embodiments of the present disclosure, crystalline form B is disclosed, having a differential scanning calorimetry curve with an endothermic peak at 249.48±8° C.

[0037] In some embodiments of the present disclosure, crystalline form B is disclosed, having a differential scanning calorimetry curve with an endothermic peak at 249.48±3° C.

[0038] In some embodiments of the present disclosure, crystalline form B is disclosed, having a DSC curve substantially as shown in FIG.

[0039] This disclosure: 1) adding a compound of formula (I) to an alcoholic solvent, ethyl acetate, tert-butyl methyl ether, tetrahydrofuran, or dichloromethane; 2) stirring at 15 to 100°C for 1 to 168 hours; 3) filtering, recovering a filter cake, and vacuum drying the filter cake at 30-45°C for 0.5-2 hours, or vacuum drying the filter cake at 35°C for 1 hour; A method for preparing crystalline form A of a compound of formula (I), wherein the compound of formula (I) has the formula (I): [ka] (I) The present invention also provides a method in which

[0040] In some embodiments of the present disclosure, the alcohol solvent is selected from methanol and ethanol.

[0041] The present disclosure also provides a use of the compound of formula (I), crystalline form A of the compound of formula (I), crystalline form B of the compound of formula (I), or a method for preparing crystalline form A of the compound of formula (I) in the manufacture of a medicament for treating an LSD1-related disease.

[0042] The present disclosure also provides a compound of formula (I), a use of crystalline form A of the compound of formula (I), or a method for preparing crystalline form A of the compound of formula (I), in the manufacture of a medicament for treating an LSD1-related disease.

[0043] The present disclosure also provides a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I), crystalline form A of the compound of formula (I), or crystalline form B of the compound of formula (I), and a pharma- ceutically acceptable carrier.

[0044] The present disclosure also provides a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I) above or crystalline form A of the compound of formula (I) above, and a pharma- ceutically acceptable carrier.

[0045] The present disclosure also provides the use of the compound of formula (I) above, the use of crystalline form A of the compound of formula (I) above, the use of crystalline form B of the compound of formula (I) above, or the use of the pharmaceutical composition above, in the manufacture of a medicament as a cardiac myosin inhibitor.

[0046] The present disclosure also provides the use of the compound of formula (I) above, the use of crystalline form A of the compound of formula (I) above, or the use of the pharmaceutical composition above, in the manufacture of a medicament as a cardiac myosin inhibitor.

[0047] The present disclosure also provides the use of the compound of formula (I) above, the use of crystalline form A of the compound of formula (I) above, the use of crystalline form B of the compound of formula (I) above, or the use of the pharmaceutical composition above, in the manufacture of a medicament for treating heart failure and hypertrophic cardiomyopathy.

[0048] The present disclosure also provides the use of the compound of formula (I) above, the use of crystalline form A of the compound of formula (I) above, or the use of the pharmaceutical composition above, in the manufacture of a medicament for treating heart failure and hypertrophic cardiomyopathy.

[0049] The present disclosure also provides a method for treating a cardiac myosin inhibitor-associated disease in a subject in need of such treatment, comprising providing to said subject an effective dose of the compound of formula (I), crystalline form A of the compound of formula (I), crystalline form B of the compound of formula (I), or a pharmaceutical composition defined in any of the above technical solutions.

[0050] The present disclosure also provides a method for treating a cardiac myosin inhibitor-associated disease in a subject in need of such treatment, comprising providing to said subject an effective dose of the compound of formula (I), crystalline form A of the compound of formula (I), or a pharmaceutical composition defined in any of the above technical solutions.

[0051] The present disclosure also provides a method for treating heart failure and hypertrophic cardiomyopathy in a subject in need thereof, comprising providing to said subject an effective dose of the compound of formula (I), crystalline form A of the compound of formula (I), crystalline form B of the compound of formula (I), or a pharmaceutical composition defined in any of the above technical solutions.

[0052] The present disclosure also provides a method for treating heart failure and hypertrophic cardiomyopathy in a subject in need thereof, comprising providing to said subject an effective dose of the compound of formula (I), crystalline form A of the compound of formula (I), or a pharmaceutical composition defined in any of the above technical solutions.

[0053] (Technical effect) The crystalline form A of the compound of formula (I) in the present disclosure is easy to obtain, has good physical and chemical stability, and has high industrial applicability and economic value. The compound of the present disclosure has good inhibitory effect on cardiac myosin ATPase and has excellent pharmacokinetic properties.

[0054] (Definitions and Explanations) Unless otherwise specified, the following terms and phrases used herein shall have the following meanings: A particular phrase or term should not be deemed unclear or indefinite due to the absence of a specific definition, but should be understood in its customary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredients.

[0055] It should be noted that, unless otherwise indicated by context or clearly contradicted by the present disclosure, in the subject matter of this disclosure (particularly the subject matter of the appended claims), as used herein and in the appended claims, the singular forms "a," "an," "the," and similar terms are to be construed as including both the singular and the plural, i.e., for example, reference to "the compound" includes reference to one or more compounds, and the like.

[0056] The term "amorphous" or "amorphous form" means that the substance, component or product does not have a characteristic crystalline shape or structure, e.g., is not substantially crystalline as determined by XRPD (X-ray powder diffraction), or that the substance, component or product is not birefringent or cubic when viewed, e.g., using a polarized light microscope, or has no sharp peaks in an X-ray powder diffraction pattern. In certain embodiments, a sample containing an amorphous form of a substance may be substantially free from other amorphous and / or crystalline forms.

[0057] Differential scanning calorimetry (DSC) of the crystalline forms of the present disclosure has experimental error and is slightly affected by the dryness of the sample. The endothermic peak position and peak value may vary slightly from machine to machine and from sample to sample. The experimental error or value difference may be 10°C or less, or 9°C or less, or 8°C or less, or 7°C or less, or 6°C or less, or 5°C or less, or 4°C or less, or 3°C or less, or 2°C or less, or 1°C or less. Therefore, the peak position or peak value of the DSC endothermic peak cannot be considered absolute.

[0058] Intermediate compounds of the present disclosure may be prepared by a variety of synthetic methods known to those of skill in the art, including the specific embodiments described below, embodiments formed by combining the specific embodiments described below with other chemical synthetic methods, and other equivalent methods known to those of skill in the art. Alternative embodiments include, but are not limited to, the examples of the present disclosure.

[0059] Unless otherwise specified, double bond structures, such as carbon-carbon double bonds, carbon-nitrogen double bonds, and nitrogen-nitrogen double bonds, are present in the compound, and each atom on the double bond is bonded to two different substituents (in a double bond containing a nitrogen atom, the unshared electron pair of the nitrogen atom is considered to be a substituent bonded thereto), and in the compound, the atoms on the double bond and their substituents have the structure: [ka] When represented by the formula: Figure US081202333-20130323-C00002 represents either the (Z)-isomer or the (E)-isomer of said compound, or a mixture of said two isomers.

[0060] The structure of the compounds disclosed herein can be determined by conventional methods known to those skilled in the art. When the present disclosure relates to the absolute configuration of a compound, its absolute configuration can be determined by conventional techniques in the art, such as single crystal X-ray diffraction (SXRD). In single crystal X-ray diffraction (SXRD), the diffraction intensity data of the grown single crystal is collected using a Bruker D8 venture diffractometer (CuKα radiation, φ / ω scan mode), and after collecting relevant data, the crystal structure is further analyzed by a direct method (ShelXs97) to determine its absolute configuration.

[0061] The chemical reactions in certain embodiments of the present disclosure are completed in suitable solvents, which must be appropriate for the chemical transformations of the present disclosure and the necessary reagents and raw materials. To obtain the compounds of the present disclosure, those skilled in the art may need to modify or select synthetic steps or reaction schemes based on existing embodiments.

[0062] The present disclosure will be described in detail below through examples, which should not be construed as limiting the present disclosure in any way.

[0063] All solvents used in this disclosure are commercially available and may be used without further purification.

[0064] The solvents used in this disclosure are commercially available.

[0065] In this disclosure, the following abbreviations are used: TEA stands for triethylamine; DIEA stands for N,N-diisopropylethylamine; PE stands for petroleum ether; EtOAc stands for ethyl acetate; EA stands for ethyl acetate; THF stands for tetrahydrofuran; MeOH stands for methanol; MTBE stands for methyl tert-butyl ether; DCM stands for dichloromethane; EtOH stands for ethanol; iPrOH stands for isopropyl alcohol; Boc2O stands for di-tert-butyl dicarbonate; L-selectride stands for lithium trisec-butylborohydride; TCFH stands for N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate; FA stands for formic acid; TFA stands for trifluoroacetic acid; AC N stands for acetonitrile; TLC stands for thin layer chromatography; HPLC stands for high performance liquid chromatography; LCMS stands for liquid chromatography-mass spectrometry; DMSO stands for dimethylsulfoxide; DMF stands for N,N-dimethylformamide; LDA stands for lithium diisopropylamide; DMAC stands for N,N-dimethylacetamide; PEG-400 stands for polyethylene glycol 400; EGTA stands for ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid; DMSO-d6 stands for deuterated dimethylsulfoxide; CDCl3 stands for deuterated chloroform; BID stands for twice a day; QD stands for once a day; PO stands for oral administration; IV stands for intravenous administration.

[0066] Compounds are named manually or using ChemDraw® software; commercially available compounds are named by their supplier directory name.

[0067] (Apparatus and analysis method in this disclosure) X-ray Powder Diffraction (XRPD) Method 1 in this disclosure: Test parameters for crystalline form A tested are shown in Table 3. Table 3 XRPD test parameters [Table 5]

[0068] X-ray powder diffraction (XRPD) method 2 in this disclosure: Data was obtained using a Bruker instrument, D2 Phaser. The test parameters are shown in Table 4. Table 4 XRPD test parameters [Table 6]

[0069] Differential Scanning Calorimetry (DSC) Method 1 in this disclosure: Test parameters are shown in Table 5. Table 5 DSC test parameters [Table 7]

[0070] Differential Scanning Calorimetry (DSC) Method 2: Test parameters in this disclosure are shown in Table 6. Table 6 DSC test parameters [Table 8]

[0071] Thermogravimetric analysis (TGA) Method 1 in this disclosure: Test parameters are shown in Table 7. Table 7 TGA test parameters [Table 9] [Brief description of the drawings]

[0072] [Figure 1] Figure 1: XRPD pattern of crystalline form A of compound of formula (I) using Cu-Kα radiation. [Diagram 2] FIG. 2 shows the DSC curve of crystalline form A of compound of formula (I). [Diagram 3] FIG. 3 shows the TGA curve of crystalline form A of compound of formula (I). [Figure 4] FIG. 4 shows an ellipsoidal drawing of the three-dimensional structure in single crystal X-ray diffraction (SC-XRD) of the compound of formula (I). [Diagram 5] FIG. 5 shows the XRPD pattern of crystalline form B of compound of formula (I) using Cu-Kα radiation. [Figure 6] FIG. 6 shows the DSC curve of crystalline form B of compound of formula (I).

[0073] (Detailed Description) In order to provide a better understanding of the present disclosure, the present disclosure is further illustrated below in conjunction with specific examples, which are not intended to limit the present disclosure. Example 1

[0074] Formula (I): [ka] (I) Synthesis Route: [ka]

[0075] Step A: 1-2 (4.66 g, 38.43 mmol, 1.2 eq) and tetraethyl titanate (21.92 g, 96.07 mmol, 19.92 mL, 3 eq) were added to a solution of 1-1 (5 g, 32.02 mmol, 4.07 mL, 1 eq) in THF (50 mL) at 20° C. The reaction solution was stirred at 60° C. for 16 hours. Ethyl acetate (100 mL) was added to the reaction solution. The mixture was cooled to 0° C., and then water (20 mL) was added slowly. The mixture was stirred for 0.5 hours and filtered. The filtrate was washed with saturated brine (50 mL×3), dried over anhydrous sodium sulfate, filtered, and then concentrated to give compound 1-3.

[0076] Step B: L-selectride (1M, 41.65 mL, 1.2 equiv) was slowly added dropwise to a solution of 1-3 (9 g, 34.71 mmol, 1 equiv) in THF (100 mL) at -78°C under nitrogen. The reaction solution was stirred at -78°C for 2 h, then slowly added to saturated aqueous ammonium chloride solution (100 mL). The mixture was extracted with EA (100 mL x 2). The combined organic layer was washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, filtered, and then concentrated. The residue was separated by column chromatography (PE: EtOAc = 5: 1 to 3: 1) to give compound 1-4.

[0077] Step C: HCl / MeOH (200 mmol, 50 mL, 7.92 equiv) was added to a solution of 1-4 (6.6 g) in MeOH (50 mL) at 20° C. The reaction solution was stirred for 16 h and then concentrated to give the hydrochloride salt of compound 1-5.

[0078] Step D: 1-6 (1.51 g, 7.74 mmol, 1.5 equiv, HCl) and DIEA (4.00 g, 30.96 mmol, 5.40 mL, 6 equiv) were added to a solution of the hydrochloride salt of 1-5 (1 g) in EtOH (10 mL) at 20° C. The reaction solution was stirred at 20° C. for 16 h and concentrated to give compound 1-7.

[0079] Step E: TEA (2.45 g, 24.24 mmol, 3.37 mL, 3 equiv) was added to a solution of 1-7 (1.54 g, 8.08 mmol, 1 equiv) in DCM (15 mL) at -20°C under nitrogen. Then, a solution of 1-8 (1.39 g, 5.14 mmol, 6.37e-1 equiv) in DCM (15 mL) was added. The reaction solution was stirred at 20°C for 16 h and then concentrated. The residue was diluted with EA (30 mL), then washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and then concentrated. The residue was separated by column chromatography (PE:EtOAc=10:1 to 5:1) to give compound 1-9.

[0080] Step F: Sodium methoxide (1M, 6.43 ml, 1 equiv) was added to a solution of 1-9 (0.648 g, 1.53 mmol, 1 equiv) in MeOH (7.6 mL) under nitrogen. The reaction solution was stirred at 20° C. for 16 h. 1M hydrochloric acid dilution was added to the reaction solution to adjust the pH to about 5, and then the mixture was extracted by adding EA (20 mL). The organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and then concentrated to give compound 1-10.

[0081] Step G: Hydrochloric acid (4M, 4mL, 17.40eq) was added to a solution of 1-10 (348mg, 919.74μmol, 1eq) in 1,4-dioxane (4mL). The reaction solution was stirred at 50°C for 16h. EA (30mL) was added to the reaction solution, then 1M aqueous sodium hydroxide was added to adjust the pH to about 8. The layers were separated. The organic layer was then washed with saturated brine (50mL x 2), dried over anhydrous sodium sulfate, filtered, and then concentrated. MeOH (10mL) was added to the residue, and the mixture was stirred for 20min and filtered. The filter cake was dried under high vacuum to obtain the compound of formula (I). After XPRD test, the obtained compound of formula (I) was crystalline form A of compound of formula (I). 1 H NMR (DMSO-d6, 400 MHz): δ ppm 9.60 - 9.47 (m, 1H), 7.34 - 7.26 (m, 2H), 7.25 - 7.16 (m, 1H), 6.96 (br d, J = 6.0 Hz, 1H), 4.82 - 4.71 (m, 1H), 4.44 - 4.34 (m, 1H), 1.93 - 1.81 (m, 4H), 1.70 (br s, 4H), 1.48 (br d, J = 6.4 Hz, 3H); LCMS(ESI) m / z: 321.2(M+1). Example 2

[0082] (Preparation of Crystalline Form A of Compound of Formula (I)) Compound of formula (I) (80 mg, 249.74 μmol) was added to ethanol (3 mL) and the mixture was stirred at 80° C. for 16 hours. The mixture was cooled to room temperature and then filtered. The solid was dried in vacuum to obtain crystalline form A of compound of formula (I). 1 H NMR (DMSO-d6, 400 MHz): δ ppm 9.60 - 9.47 (m, 1H), 7.34 - 7.26 (m, 2H), 7.25 - 7.16 (m, 1H), 6.96 (br d, J = 6.0 Hz, 1H), 4.82 - 4.71 (m, 1H), 4.44 - 4.34 (m, 1H), 1.93 - 1.81 (m, 4H), 1.70 (br s, 4H), 1.48 (br d, J = 6.4 Hz, 3H); LCMS(ESI) m / z: 321.2(M+1). The XRPD, DSC and TGA of the crystalline form A of compound of formula (I) were all tested using the corresponding method 1, and the test result patterns and curves are shown in Figures 1, 2 and 3, respectively.

[0083] By replacing the solvent ethanol with methanol, dichloromethane, ethyl acetate, or tetrahydrofuran, crystalline form A of the compound of formula (I) was obtained at room temperature or at elevated temperature in each case. The assay results are shown in Table 8. Table 8. Assay of crystalline forms obtained by stirring in different solvents [Table 10] Example 3

[0084] (Preparation of Crystalline Form B of Compound of Formula (I)) Approximately 60 mg of the crystalline form A was weighed out and heated from room temperature (20-25°C) to 250°C at a rate of 10°C / min using a TGA, and then naturally cooled to room temperature (the cooling rate was not controlled during the cooling process) to obtain crystalline form B of the compound of formula (I). 1H NMR (DMSO-d6, 400 MHz): δ ppm 9.57 (s, 1H), 7.32 - 7.27 (m, 2H), 7.22 - 7.15 (m, 1H), 6.96 (br d, J = 6.4 Hz, 1H), 4.78 - 4.72 (m, 1H), 4.38 (s, 1H), 1.86 - 1.82 (m, 4H), 1.70 - 1.67 (m, 4H), 1.47 (br d, J = 6.8 Hz, 3H). XRPD and DSC of crystalline form B of compound of formula (I) were both tested using the corresponding method 2, and the test pattern and curve are shown in Figures 5 and 6, respectively. Example 4

[0085] (Single crystal X-ray diffraction analysis of the compound of formula (I)) 1. Instrument Parameters and Data Acquisition Manufacturer:Bruker Corporation; Equipment model: Bruker D8 VENTURE; X-ray source: A high-intensity microfocus rotating anode light source was used; Cu rotating target: λ=1.54184Å; Power: 2.5kw; Tube voltage: 50kV; Tube current: 50mA; Goniometer: 4-axis (Kappa, ω, 2θ, φ) goniometer; Detector: Large area photon II detector, effective detector area 14cm x 10cm Detector to sample distance: d = 45 mm.

[0086] 2.Crystal Growth A sample of 14.3 mg was weighed and added to MeOH (2 mL). The sample was insoluble. Then DMSO (1 mL) was added dropwise and the mixture was stirred until completely dissolved. The dissolved sample was transferred to a 4 mL semi-sealed sample vial and slowly evaporated at room temperature. After 7 days, colorless block crystals were obtained and submitted for single crystal testing.

[0087] 3. Crystal Data Table Table 9. Single crystal structure data for compound of formula (I) [Table 11]

[0088] 4. Conclusion The single crystal data indicated that the single crystal was the compound of formula (I). An ellipsoidal view of the three-dimensional structure of the single crystal X-ray diffraction (SC-XRD) of the compound of formula (I) is shown in Figure 4. The single crystal structural data and parameters of the compound of formula (I) are shown in Table 9.

[0089] Biological Assay Data: Assay Example 1: Assay of inhibitory effect on cardiac myosin ATPase activity Assay Reagents: Cardiac tropomyosin / troponin complex (Cytoskeleton, Cat.#TT05) Cardiac myosin S1 (Cytoskeleton, Cat.#MYS03) Cardiac actin (Cytoskeleton, Cat.#AD99-A) ATPase assay biochem kit (Cytoskeleton, Cat.#BK051)

[0090] Assay steps: 1) Preparation of compounds a) The compound was serially diluted in 8 steps of 4-fold concentration with DMSO in Echo, and 200 nL of each concentration of the compound was transferred to a 96-well plate (Corning-3696). b) The plate was centrifuged at 1000 rpm for 15 seconds and sealed for later use. 2) Preparation of F-actin a) 5 mM Pipes-KOH buffer (pH 7.0), 500 μM ATP, and 500 μM dithiothreitol were prepared, and 2.5 mL of the buffer was added thereto, and 1 mg of F-actin was dissolved therein to give a protein concentration of 0.4 mg / mL. b) The mixture was allowed to stand at room temperature for 10 minutes to completely dissolve the protein. c) 2.0 mM MgCl2 and 2.0 mM EGTA were added and the mixture was allowed to stand at room temperature for 20 minutes to allow protein polymer formation. 3) Preparation of thin filaments a) 200 μL of ice water was added to dissolve 1 mg of cardiac tropomyosin / troponin complex, resulting in a protein concentration of 5 mg / mL. b) 1000 μL of F-actin prepared in step 1 was added and mixed well. c) The mixture was allowed to stand at room temperature for 20 minutes. d) The mixture was centrifuged at 87K×g at 4° C. for 1.5 hours. e) PM12 buffer (12 mM Pipes-KOH, pH 7.0, 2 mM MgCl2) was prepared and 1200 μL of the buffer was added to resuspend the protein. 4) Preparation of reaction solutions and initiation of assay a) 250 μL of ice-cold PM12 buffer was added to 250 μg of S1 myosin to give a protein concentration of 1 mg / mL. b) The reagents were added sequentially in the following order and mixed to obtain a reaction mixture: 400 μL PM12, 400 μL 5×MSEG (from the biochem kit for the ATPase assay), 1200 μL of actin / cardiac tropomyosin / troponin complex, 40 μL of myosin S1, 40 μL of 100× PNP (from the biochem kit for the ATPase assay), 10.4 μL of 100 mM ATP. c) 10 μL of 440 μM CaCl 2 solution was added to the 96-well plate, and the plate was transferred to a 37° C. incubator to pre-heat. d) 100 μL of the reaction mixture was added to the 96-well plate, and the plate was centrifuged at 1000 rpm for 10 seconds. e) The plate was read continuously for 10 minutes at 30 second intervals on a SpectraMax340PC with an instrument temperature of 37° C. and a wavelength of 360 nm.

[0091] Data Analysis: Data was analyzed using Prism and the assay results are shown in Table 10. Table 10. Assay Results: IC of the compounds disclosed herein, which are inhibitory effects on cardiac myosin ATPase activity. 50 value [Table 12] Conclusion: The compounds of the present disclosure have good inhibitory activity against cardiac myosin ATPase.

[0092] Assay Example 2: Pharmacokinetic evaluation in rats Purpose of the assay: The pharmacokinetic parameters of compounds of the present disclosure were determined in rats. Assay scheme: 1) Assay Agent: a compound of the present disclosure; 2) Assay animals: Four male SD rats (7–9 weeks old) were randomly divided into two groups, each containing two rats; 3) Drug preparation: An appropriate amount of the drug was weighed and dissolved in a mixed solvent (DMAC:PEG-400:2-HP-β-CD (30%) = 5:25:70, adjusted to 0.2 mg / mL); Assay procedure: Animals in group 1 were administered the drug at a dose of 0.2 mg / kg per tail vein injection at a concentration of 0.2 mg / mL. Animals in group 2 were administered the compound at a dose of 1 mg / kg per tail vein injection at a concentration of 0.2 mg / mL by force-feeding. Plasma samples were collected from the animals at 0.0833 (tail vein injection group only), 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration. Data Analysis: Drug concentrations in plasma samples were determined using LC-MS / MS methodology. The pharmacokinetic assay results obtained for the assayed drugs are shown in Table 11. Table 11. Pharmacokinetic assay results for compounds of the present disclosure [Table 13] -- indicates no data available. Conclusion: The compounds of the present disclosure have good pharmacokinetic properties in rats.

Claims

1. Characterized by an X-ray powder diffraction pattern (XPRD) having characteristic diffraction peaks at angles 2θ of 18.283±0.200°, 19.662±0.200°, and 22.420±0.200°, Formula (I): 【Chemistry 1】 (I) Crystalline form A of the compound of formula (I).

2. 2. The crystalline form A of claim 1, characterized by an X-ray powder diffraction pattern (XPRD) having characteristic diffraction peaks at angles 2θ of 11.143±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 26.259±0.200°, and 28.056±0.200°.

3. 3. The crystalline form A of claim 2, characterized by an X-ray powder diffraction pattern having characteristic diffraction peaks at angles 2θ of 11.143±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 26.259±0.200°, 27.261±0.200°, 28.056±0.200°, and 30.256±0.200°.

4. 4. The crystalline form A of claim 3, characterized by an X-ray powder diffraction pattern having characteristic diffraction peaks at angles 2θ of 11.143±0.200°, 16.522±0.200°, 17.053±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 23.909±0.200°, 26.259±0.200°, 27.261±0.200°, 28.056±0.200°, 30.256±0.200°, and 33.761±0.200°.

5. 2. The crystalline form A of claim 1, characterized by an X-ray powder diffraction pattern comprising at least 5, 6, 7, or 8 characteristic diffraction peaks representative of angles 2θ selected from the following angles: 11.143±0.200°, 18.283±0.200°, 19.662±0.200°, 22.420±0.200°, 26.259±0.200°, 27.261±0.200°, 28.056±0.200°, and 30.256±0.200°.

6. 11.143°, 13.260°, 14.860°, 16.163°, 16.522°, 17.053°, 17.537°, 18.283°, 19.662°, 22.420°, 23.568°, 23.909°, 26.259°, 26.726°, 27.261°, 28.056°, 29.008°, 30.256° 6. The crystalline form A of claim 5, characterized by an X-ray powder diffraction pattern having characteristic diffraction peaks at angles 2θ of 31.219°, 31.646°, 32.037°, 32.438°, 32.807°, 33.761°, 34.534°, 35.404°, 36.856°, 37.813°, and 39.456°.

7. 2. The crystalline form A of claim 1, having an XRPD pattern substantially as shown in FIG.

8. The crystalline form A of any one of claims 1 to 7, having a differential scanning calorimetry (DSC) curve with endothermic peak values ​​at 249.53±3°C and 306.12±3°C.

9. 2. The crystalline form A of claim 1, having a DSC curve substantially as shown in FIG.

10. The crystalline form A according to any one of claims 1 to 7, having a thermogravimetric analysis (TGA) curve with a maximum weight loss of 0.075% at 200±3°C and a maximum weight loss of 0.164% at 250±3°C.

11. 2. The crystalline form A of claim 1, having a TGA curve substantially as shown in FIG.

12. 1) adding a compound of formula (I) to an alcohol solvent, ethyl acetate, tert-butyl methyl ether, tetrahydrofuran, or dichloromethane; 2) Stirring at 15 to 100° C. for 1 to 168 hours; 3) filtering, recovering the filter cake, and vacuum drying the filter cake at 30-45°C for 0.5-2 hours, or vacuum drying the filter cake at 35°C for 1 hour; 2. A method for preparing crystalline form A of the compound of formula (I) according to claim 1, wherein the compound of formula (I) has the formula (I): 【Chemistry 2】 (I) That is, the method.

13. A pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) according to any one of claims 1 to 11 or crystalline form A of compound of formula (I) and a pharma- ceutically acceptable carrier.

14. Use of a compound of formula (I) as defined in any one of claims 1 to 11 or crystalline form A of compound of formula (I), or a pharmaceutical composition as defined in claim 13, in the manufacture of a medicament as a cardiac myosin inhibitor.

15. Use of a compound of formula (I) or crystalline form A of compound of formula (I) as defined in any one of claims 1 to 11, or a pharmaceutical composition as defined in claim 13, in the manufacture of a medicament for the treatment of heart failure and hypertrophic cardiomyopathy.

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