Crystal forms of pyridine-3-carboxylic acid 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl ester and methods of synthesis

Abstract

The present invention provides polymorphs of a compound of Formula (X). The present invention also provides pharmaceutical compositions containing the polymorphs of the compound and methods of treating a condition in a subject by providing the polymorphs of the compound.

Description

[0001] Cross-reference to related applications

[0002] This application claims the benefit and priority of U.S. Provisional Patent Application No. 63 / 046,120, filed June 30, 2020, and U.S. Provisional Patent Application No. 63 / 046,123, filed June 30, 2020, the contents of each of which are incorporated herein by reference. Technical Field

[0003] This invention relates to the crystallographic form of 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylic acid and a method for the chemical synthesis of the compound. Background Technology

[0004] Heart disease is the leading cause of death worldwide, claiming 15 million lives globally in 2015. In many forms of heart disease, reduced cardiac efficiency is caused by changes in mitochondrial energy metabolism. Mitochondria are subcellular compartments where metabolites derived from glucose and fatty acids are oxidized to produce high-energy molecules. Increased fatty acid oxidation in the heart reduces glucose oxidation, and vice versa. Glucose oxidation is a more efficient energy source, but in some types of heart disease (such as heart failure, ischemic heart disease, and diabetic cardiomyopathy), fatty acid oxidation dominates in the cardiac mitochondria. Consequently, the heart's pumping capacity is reduced.

[0005] CV-8972, with the IUPAC name 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylic acid, has the following structure:

[0006]

[0007] Due to its pharmacokinetic characteristics, it has recently been identified as a promising therapeutic candidate for the treatment or prevention of cardiac symptoms. Summary of the Invention

[0008] This document provides crystallographic forms of CV-8972 and compositions containing these crystallographic forms. The present invention recognizes that CV-8972 crystals exist in a variety of polymorphic forms, and that polymorphic form A is the most stable under ambient temperature and relative humidity conditions. Therefore, CV-8972 crystals in form A can be used to manufacture pharmaceutical compositions. For example, pharmaceutical compositions containing polymorph A do not require special treatment during storage or dispensing. Furthermore, such compositions retain their efficacy better than compositions containing other polymorphs or mixtures of polymorphs. The present invention also provides a method for treating cardiac symptoms in a subject using CV-8972 polymorphs, such as form A.

[0009] This invention also provides a method for synthesizing CV-8972. Previous methods for synthesizing CV-8972 required the formation of a free base form of 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethanol, also known as CV-8814, and the conversion of this free base form of CV-8814 into its hydrochloride salt. In such methods, CV-8814 must then be converted back to its free base form for coupling with nicotinic acid to form the free base form of CV-8972. This invention provides a synthetic method for CV-8972 that bypasses the reversible conversion of CV-8814 between its free base and HCl salt forms. In the method provided herein, the free base form of CV-8814 is formed in a reductive amination reaction, and the free base product is used directly as a substrate for coupling with nicotinic acid to form CV-8972. Due to the fewer steps required, the synthetic method of this invention is simpler, faster, and provides better yields than previous methods for preparing CV-8972.

[0010] On one hand, the present invention provides crystals comprising polymorphs of compounds of formula (X):

[0011]

[0012] The polymorph can be form A, form B, form C, form D, or form E.

[0013] The crystal may be substantially free of one or more other polymorphs. For example, the crystal may contain polymorph A and be substantially free of polymorphs B, C, D and E.

[0014] The crystal may contain the hydrochloride salt of compound (X). The crystal may contain compound (X) and hydrochloride ions in a defined stoichiometric ratio. The crystal may contain the compound and hydrochloride ions in a stoichiometric ratio of 1:3.

[0015] The crystal may contain the hydrated form of the compound of formula (X). The crystal may contain the monohydrate form of the compound. The crystal may contain the anhydrous form of the compound.

[0016] On the other hand, the present invention provides a pharmaceutical composition comprising a polymorph of a compound of formula (X):

[0017] The polymorph can be form A, form B, form C, form D, or form E.

[0018] The composition may be substantially free of one or more other polymorphs. For example, the composition may contain polymorph A and be substantially free of polymorphs B, C, D and E.

[0019] The composition may contain the hydrochloride salt of compound (X). The composition may contain compound (X) and hydrochloride ions in a defined stoichiometric ratio. The composition may contain the compound and hydrochloride ions in a stoichiometric ratio of 1:3.

[0020] The composition may comprise the hydrated form of the compound of formula (X). The composition may comprise the monohydrate form of the compound. The composition may comprise the anhydrous form of the compound.

[0021] This composition can be formulated for any route or mode of administration. It can be formulated for oral, skin, enteral, intra-arterial, intramuscular, intraocular, intravenous, nasal, oral, parenteral, pulmonary, rectal, subcutaneous, topical, or percutaneous application. It can also be formulated for administration by injection or with or on an implantable medical device (e.g., a stent, or a drug-eluting stent or balloon equivalent).

[0022] The composition can be formulated into a single unit dose. The composition can be formulated into separate doses.

[0023] The composition may contain a defined dose of the compound. The dose may contain about 10 mg to about 2000 mg, about 10 mg to about 1000 mg, about 10 mg to about 800 mg, about 10 mg to about 600 mg, about 10 mg to about 400 mg, about 10 mg to about 300 mg, about 10 mg to about 200 mg, about 25 mg to about 2000 mg, about 25 mg to about 1000 mg, about 25 mg to about 800 mg, about 25 mg to about 600 mg, about 25 mg to about 400 mg, about 25 mg to about 300 mg, about 25 mg to about 200 mg, about 50 mg to about 2000 mg, about 50 mg to about 1000 mg, about 50 mg to about 800 mg, about 50 mg to about 600 mg, about 50 mg to about 400 mg, about 50 mg to about 300 mg, about... The compound in the form of 50 mg to about 200 mg, about 100 mg to about 2000 mg, about 100 mg to about 1000 mg, about 100 mg to about 800 mg, about 100 mg to about 600 mg, about 100 mg to about 400 mg, about 100 mg to about 300 mg, about 100 mg to about 200 mg, about 200 mg to about 2000 mg, about 200 mg to about 1000 mg, about 200 mg to about 800 mg, about 200 mg to about 600 mg, about 200 mg to about 400 mg, about 200 mg to about 300 mg, about 300 mg to about 2000 mg, about 300 mg to about 1000 mg, about 300 mg to about 800 mg, about 300 mg to about 600 mg, or about 300 mg to about 400 mg. The dosage may contain about 10 mg, about 25 mg, about 50 mg, about 100 mg, about 200 mg, about 300 mg, or about 400 mg of the compound.

[0024] The composition may contain crystals of a compound of formula (X). The crystals may possess any of the properties described above for crystals of the compound.

[0025] On the other hand, the present invention provides a method for treating a subject’s condition by providing a composition to a subject who has or is at risk of developing the condition, the composition containing a therapeutically effective amount of a polymorph of a compound of formula (X).

[0026] The polymorph can be form A, form B, form C, form D, or form E.

[0027] The composition may have any of the properties described above for the composition containing the compound of formula (X) (a crystal containing the compound).

[0028] The composition can be delivered via any suitable route or mode of administration. It can be delivered orally, through the skin, intestines, arteries, muscles, eyes, veins, nose, orally, parenterally, lungs, rectum, subcutaneously, topically, percutaneously, by injection, or via or onto an implantable medical device (e.g., a stent, or a drug-eluting stent or balloon equivalent).

[0029] This composition can be provided as a single unit dose. This composition can also be provided as separate doses.

[0030] This composition may be provided in one dose per day. This composition may be provided in multiple doses per day. This composition may be provided in two, three, four, five, six, eight or more doses per day.

[0031] The composition may contain a defined dose of the compound, such as any of the doses described above.

[0032] The one or more doses may be provided over a defined period of time. The one or more doses may be provided daily for at least one week, at least two weeks, at least three weeks, at least four weeks, at least six weeks, at least eight weeks, at least ten weeks, at least twelve weeks, or longer.

[0033] This condition can be a cardiovascular condition. Such conditions include aneurysm, angina pectoris, atherosclerosis, cardiomyopathy, cerebrovascular disease, congenital heart disease, coronary artery disease, coronary heart disease, diabetic cardiomyopathy, heart attack, heart disease, heart failure, hypertension, ischemic heart disease, pericardial disease, peripheral artery disease, rheumatic heart disease, stroke, transient ischemic attack, or valvular heart disease. This angina pectoris may be refractory to other medical interventions.

[0034] This condition can be a rheumatic condition. Rheumatic conditions can include acute kidney injury, alcoholic cardiomyopathy, angina pectoris (e.g., refractory angina and heart failure-related angina), ankylosing spondylitis, autoimmune-associated lung disease, Behcet's disease, bursitis, cachexia, myocardial fibrosis, chemotherapy-induced chronic fatigue syndrome, claudication (e.g., peripheral claudication), contrast-induced nephropathy, cyanotic cardiomyopathy, dermatomyositis, dilated cardiomyopathy, imbalance, fibromyalgia, weakness, gout, and Gulf War syndrome. Syndrome), heart failure, hypertrophic cardiomyopathy, induced nephropathy, infectious arthritis, inflammatory arthritis, inflammatory eye disease, inflammatory myopathy, ischemic cardiomyopathy, juvenile idiopathic arthritis, left ventricular failure, lupus, myofascitis, myofascial pain syndrome, myositis, osteoarthritis, osteonecrosis of the jaw, osteoporosis, polymyalgia rheumatica, polymyositis, psoriatic arthritis, pulmonary hypertension, pulmonary fibrosis, rare muscle diseases, rheumatoid arthritis, sarcoidosis, sarcopenia, scleroderma, Sjogren's syndrome, tendinitis, tinnitus, vasculitis, or vertigo.

[0035] The condition can be fibrosis. This fibrosis may be associated with another disease, condition, or symptom. For example, the fibrosis may include or be associated with the following: adhesive capsulitis, aneurysm, angina pectoris, arteriosclerosis, joint fibrosis, atherosclerosis, atrial fibrosis, cardiomyopathy, cerebrovascular disease, cirrhosis, congenital heart disease, coronary artery disease, coronary heart disease, Crohn's disease, cystic fibrosis, diabetic cardiomyopathy, Dupuytren's contracture, endocardial fibrosis, glial scars, heart attack, heart failure, high blood pressure / hypertension, idiopathic pulmonary fibrosis, ischemic heart disease, keloids, mediastinal fibrosis, myelofibrosis, renal systemic fibrosis, old myocardial infarction, pericardial disease, peripheral artery disease, Peyronie's disease. Diseases including: progressive massive fibrosis, pulmonary fibrosis, radiation-induced lung injury, retroperitoneal fibrosis, rheumatic heart disease, scleroderma, stroke, systemic sclerosis, transient ischemic attack, or valvular heart disease.

[0036] This condition can be cancer. The cancer can be bladder cancer, brain cancer, breast cancer, carcinoma, cervical cancer, colon cancer, colorectal cancer, stomach cancer, glioblastoma, glioma, head and neck cancer, kidney cancer, leukemia, liposarcoma, liver cancer, lung cancer, lymphoma, medullablastoma, melanoma, muscle cancer, neuroblastoma, oligodendroastrocytoma, oligodendroglioma, osteosarcoma, ovarian cancer, pancreatic cancer, paraganglioma, prostate cancer, sarcoma, or thyroid cancer.

[0037] On the other hand, the present invention provides a method for altering cardiac remodeling by providing a composition to a subject who has or is at risk of developing cardiac remodeling, the composition containing a therapeutically effective amount of a polymorph of a compound of formula (X).

[0038] The polymorph can be form A, form B, form C, form D, or form E.

[0039] The composition may have any of the properties described above for the composition containing the compound of formula (X) (a crystal containing the compound).

[0040] This composition can be delivered via any suitable route or mode of administration. It can be delivered orally, through the skin, enterally, intraocularly, intravenously, nasally, orally, parenterally, pulmonaryly, subcutaneously, topically, percutaneously, by injection, or through or onto an implantable medical device (e.g., a stent, or a drug-eluting stent or balloon equivalent).

[0041] This composition can be provided as a single unit dose. This composition can also be provided as separate doses.

[0042] This composition may be provided in one dose per day. This composition may be provided in multiple doses per day. This composition may be provided in two, three, four, five, six, eight or more doses per day.

[0043] The composition may contain a defined dose of the compound, such as any of the doses described above.

[0044] The one or more doses may be provided over a defined period of time. The one or more doses may be provided daily for at least one week, at least two weeks, at least three weeks, at least four weeks, at least six weeks, at least eight weeks, at least ten weeks, at least twelve weeks, or longer.

[0045] This cardiac remodeling may be associated with disease, condition, or symptom. It may be related to cardiovascular disease. For example, this cardiac remodeling may be associated with: subclavian artery anomaly, aortic regurgitation, aortic stenosis, arteriovenous malformations and fistulas, atrial septal defect, atrioventricular septal defect, bicuspid aortic valve, cardiac hypertrophy, cardiomyopathy, aortic coarctation, complete heart block, concentric hypertrophy, congenital heart defects, congenital heart disease, coronary artery disease, dextrocardia, right rotation of the great arteries, diabetes, dieting, double aortic arches, double inlet of the left ventricle, double outlet of the right ventricle, Ebstein's anomaly, giant hepatic hemangioma, heart failure, high cholesterol, high-output hemodialysis fistula, hypertension, left ventricular hypoplasia syndrome, right ventricular hypoplasia syndrome, aortic arch transection, left rotation of the great arteries, mitral regurgitation, which also causes left atrial volume overload, mitral stenosis, myocardial ischemia, obesity, outlet obstruction, partial anomalous pulmonary vein connection, patent ductus arteriosus, and Cantrell's pentalogy of Cantrell syndrome, persistent truncus arteriosus, pressure overload, pulmonary atresia, pulmonary hypertension, pulmonary valve regurgitation, pulmonary stenosis, rhabdomyosarcoma, right ventricular volume overload, scimitar syndrome, Shone's syndrome, tetralogy of Fallot, complete pulmonary venous connection abnormality, transposition of the great vessels, tricuspid atresia, tricuspid regurgitation, use of tobacco, alcohol or other drugs, valvular heart disease, ventricular dilatation, ventricular hypertrophy, ventricular septal defect, volume overload, and Wolff-Parkinson-White syndrome.

[0046] On the other hand, the present invention provides the use of crystals containing polymorphs of compounds of formula (X) for the preparation of pharmaceuticals.

[0047] In embodiments of the application, the polymorph is form A, form B, form C, form D, or form E.

[0048] In embodiments intended for use, the crystal is substantially free of one or more other polymorphs. In embodiments intended for use, the crystal comprises polymorph A and is substantially free of polymorphs B, C, D, and E.

[0049] In embodiments of use, the crystal comprises the hydrochloride salt of compound (X). In embodiments of use, the crystal comprises compound (X) and chloride ions in a defined stoichiometric ratio. In embodiments of use, the crystal comprises the compound and chloride ions in a 1:3 stoichiometric ratio.

[0050] In embodiments of use, the drug comprises a hydrated form of the compound of formula (X). In embodiments of use, the drug comprises a monohydrate form of the compound. In embodiments of use, the drug comprises an anhydrous form of the compound.

[0051] In embodiments of use, the drug is formulated for oral, skin, enteral, intra-arterial, intramuscular, intraocular, intravenous, nasal, oral, parenteral, pulmonary, rectal, subcutaneous, topical, or transdermal administration. In embodiments of use, the drug is formulated for administration by injection or onto an implantable medical device (e.g., a stent, or a drug-eluting stent or balloon equivalent).

[0052] In one embodiment of use, the drug is formulated into a single unit dose. In another embodiment of use, the drug is formulated into separate doses.

[0053] In embodiments of use, the drug contains about 10 mg to about 2000 mg, about 10 mg to about 1000 mg, about 10 mg to about 800 mg, about 10 mg to about 600 mg, about 10 mg to about 400 mg, about 10 mg to about 300 mg, about 10 mg to about 200 mg, about 25 mg to about 2000 mg, about 25 mg to about 1000 mg, about 25 mg to about 800 mg, about 25 mg to about 600 mg, about 25 mg to about 400 mg, about 25 mg to about 300 mg, about 25 mg to about 200 mg, about 50 mg to about 2000 mg, about 50 mg to about 1000 mg, about 50 mg to about 800 mg, about 50 mg to about 600 mg, about 50 mg to about 400 mg, about 50 mg to about 300 mg. The compound in g, about 50 mg to about 200 mg, about 100 mg to about 2000 mg, about 100 mg to about 1000 mg, about 100 mg to about 800 mg, about 100 mg to about 600 mg, about 100 mg to about 400 mg, about 100 mg to about 300 mg, about 100 mg to about 200 mg, about 200 mg to about 2000 mg, about 200 mg to about 1000 mg, about 200 mg to about 800 mg, about 200 mg to about 600 mg, about 200 mg to about 400 mg, about 200 mg to about 300 mg, about 300 mg to about 2000 mg, about 300 mg to about 1000 mg, about 300 mg to about 800 mg, about 300 mg to about 600 mg, or about 300 mg to about 400 mg. In embodiments of use, the drug contains about 10 mg, about 25 mg, about 50 mg, about 100 mg, about 200 mg, about 300 mg, or about 400 mg of the compound.

[0054] On the other hand, the present invention provides a method for preparing compounds of formula (X):

[0055]

[0056] This preparation is carried out by performing the following steps:

[0057] React 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol to produce the free base form of compound (IX):

[0058] as well as

[0059] The free base form of compound (IX) is reacted with nicotinic acid to produce compound (X).

[0060] This method does not include the salt form that produces the compound of formula (IX).

[0061] The step of reacting 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol-1-ol may include one or more solvents, catalysts, or other chemicals. The step of reacting 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol-1-ol may include one or more of sodium triacetoxyborohydride, acetic acid, and 2-methyltetrahydrofuran.

[0062] The reaction of 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol-1-ol can be carried out at a defined temperature. The reaction can be carried out at approximately 10°C to approximately 30°C, approximately 15°C to approximately 30°C, approximately 20°C to approximately 30°C, approximately 25°C to approximately 30°C, approximately 10°C to approximately 25°C, approximately 15°C to approximately 25°C, approximately 20°C to approximately 25°C, approximately 10°C to approximately 20°C, or approximately 15°C to approximately 20°C.

[0063] The step of reacting 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol-1-ol may not involve a specific solvent, catalyst, or other chemical. The step of reacting 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol-1-ol may not involve dichloromethane.

[0064] The step of reacting the free base form of compound (IX) with nicotinic acid can produce the free base form of compound (X).

[0065] The step of reacting the free base form of compound (IX) with nicotinic acid may include one or more solvents, catalysts, or other chemicals. The step of reacting the free base form of compound (IX) with nicotinic acid may include one or more of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, 4-(dimethylamino)pyridine, and dichloromethane.

[0066] The step of reacting the free base form of compound (IX) with nicotinic acid can be carried out at a defined temperature. The step of reacting the free base form of compound (IX) with nicotinic acid can be carried out at about 15°C to about 30°C, about 20°C to about 30°C, about 25°C to about 30°C, about 15°C to about 25°C, about 20°C to about 25°C, or about 15°C to about 20°C.

[0067] The method may include converting the free base form of compound (X) into a salt form of compound (X). The salt form of compound (X) may be an HCl salt. The salt form of compound (X) may be a monohydrate.

[0068] The step of converting the free base form of compound (X) to the salt form of compound (X) may include one or more solvents, catalysts, or other chemicals. The step of converting the free base form of compound (X) to the salt form of compound (X) may include one or more of HCl and methyl ethyl ketone.

[0069] The step of converting the free base form of compound (X) to the salt form of compound (X) can be carried out at a defined temperature. The step of converting the free base form of compound (X) to the salt form of compound (X) can be carried out at approximately 40°C to approximately 60°C, approximately 45°C to approximately 60°C, approximately 50°C to approximately 60°C, approximately 55°C to approximately 60°C, approximately 40°C to approximately 55°C, approximately 45°C to approximately 55°C, approximately 50°C to approximately 55°C, approximately 40°C to approximately 50°C, approximately 40°C to approximately 50°C, approximately 40°C, approximately 45°C, approximately 50°C, approximately 55°C, or approximately 60°C.

[0070] The method may include converting the salt form of the compound of formula (X) from a first crystalline form to a second crystalline form. Each of the first crystalline form and the second crystalline form may independently be form A, form B, form C, form D, or form E.

[0071] The step of converting the salt form of compound (X) from the first crystalline form to the second crystalline form may include one or more of the following: changing the solvent of the salt form of compound (X) and incubating the salt form of compound (X) at about 60°C.

[0072] This method can be carried out without the use of one or more solvents, catalysts, or other chemicals. This method can also be carried out without the use of one or more of dioxane, ethyl acetate, or potassium carbonate.

[0073] The method may include purifying the free base form of the compound of formula (IX). The method may also include crystallizing the free base form of the compound of formula (IX).

[0074] On the other hand, the present invention provides a method for preparing a compound of formula (X) by performing the following steps:

[0075] Make the compound of formula (1):

[0076]

[0077] Compounds of formula (2):

[0078]

[0079] The reaction produces a compound of formula (IX) in the form of a free base:

[0080]

[0081] Make the free base form of compound (IX) the same as that of compound (3):

[0082]

[0083] The reaction produces a compound of formula (X) in its free base form; and

[0084] The free base form of compound (X) is converted into the HCl salt of compound (X).

[0085] This method does not include the salt form that produces the compound of formula (IX).

[0086] The method may include purifying the free base form of the compound of formula (IX). The method may also include crystallizing the free base form of the compound of formula (IX). Attached Figure Description

[0087] Figure 1 It is a space-filled three-dimensional model of the crystal structure of the D polymorph of CV-8972.

[0088] Figure 2 It is a space-filled three-dimensional model of the crystal structure of the D polymorph of CV-8972 at room temperature.

[0089] Figure 3It is a space-filled three-dimensional model of the crystal structure of polymorph A in the form of CV-8972.

[0090] Figure 4 This is the XRPD diffraction pattern of the CV-8972 starting material.

[0091] Figure 5 The TGA and DSC thermograms of the CV-8972 starting material are shown.

[0092] Figure 6 Various forms of XRPD diffraction patterns of CV-8972 are shown.

[0093] Figure 7 This is a polarization micrograph of the starting material of CV-8972.

[0094] Figure 8 It is a dynamic vapor adsorption isotherm diagram.

[0095] Figure 9 The XRPD diffraction patterns of CV-8972 before and after dynamic vapor adsorption are shown.

[0096] Figure 10 The XRPD diffraction patterns of CV-8972 in its dehydrated and rehydrated forms are shown. Figure 11 XRPD diffraction patterns of various polymorphs of CV-8972 are shown.

[0097] Figure 12 It is C 22 H 34 A series of PLM images of single crystals of Cl3N3O6 (CV-8972).

[0098] Figure 13 A PLM image of the crystal used in a single-crystal diffractometer is shown.

[0099] Figure 14 An image of a crystal mounted on a 100-micron Mitegen ring is shown.

[0100] Figure 15 It is C 22 H 34 Ortep diagram of the asymmetric unit of Cl3N3O6 crystal.

[0101] Figure 16 C is shown 22 H 34 A unit cell of Cl3N3O6 crystal.

[0102] Figure 17 It is C 22 H 34 Diagram of hydrogen bond network and counter ion pairs in Cl3N3O6 crystal.

[0103] Figure 18 C is shown 22 H 34 XRPD plot of Cl3N3O6 crystals calculated and measured.

[0104] Figure 19 A PLM image of a single crystal-free body from recrystallized CV-8972 is shown.

[0105] Figure 20 This is an image of a single anhydrous crystal from recrystallized CV-8972 mounted on a glass fiber tip.

[0106] Figure 21 It is C 22 H 32 Thermal ellipsoid diagram of the asymmetric unit of Cl3N3O5 crystal.

[0107] Figure 22 C is shown 22 H 32 A unit cell of Cl3N3O5 crystal.

[0108] Figure 23 It is C 22 H 32 Diagram of hydrogen bond network and counter ion pairs in Cl3N3O5 crystal.

[0109] Figure 24 C is shown 22 H 34 XRPD plot of Cl3N3O6 crystals calculated and measured. Detailed Implementation

[0110] The recently identified compound CV-8972 shows promise as a therapeutic agent for a variety of conditions, including cardiovascular disease, rheumatic diseases, fibrosis, and cancer. CV-8972, with the IUPAC name 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylic acid, has the following structure:

[0111]

[0112] It is metabolized in vivo into two sets of products, which increase mitochondrial energy production in different ways. In the initial reaction, the molecule splits into CV-8814, which has the following structure:

[0113]

[0114] And niacin. Over time, CV-8814 is converted to trimetazidine in the body. Both CV-8814 and trimetazidine inhibit fatty acid β-oxidation, thus shifting mitochondrial metabolism towards glucose oxidation, a more oxygen-efficient energy source. Niacin is used to synthesize nicotinamide adenine dinucleotide (NAD). + The precursor to NAD. + It promotes mitochondrial respiration to drive ATP synthesis, regardless of whether glucose or fatty acids are used as a carbon source. Therefore, the two sets of products generated from the breakdown of CV-8972 in vivo work synergistically to stimulate mitochondria in cardiac tissue and other cell types to produce energy. CV-8972 and its mechanism of action are described in U.S. Patent No. 10,556,013, the contents of which are incorporated herein by reference.

[0115] U.S. Patent No. 10,556,013 also provides a scheme for the synthesis of CV-8972. This scheme requires the reductive amination of 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethanol-1-ol to form the free base form of CV-8814. Since it is difficult to isolate CV-8814 in solid form in this prior method, the product of the reaction is subsequently converted to the hydrochloride salt of CV-8814. However, CV-8814 must be converted back to its free base form for esterification with nicotinic acid to produce CV-8972.

[0116] The present invention recognizes that CV-8972 crystals exist in a variety of polymorphic forms. One polymorph, form A, is most stable under ambient temperature and relative humidity conditions, and therefore has specific utility for the manufacture of pharmaceutical compositions. Due to the stability of form A, compositions containing this polymorph can be easily stored and dispensed without loss of therapeutic efficacy. Therefore, the present invention provides compositions containing polymorphs of crystalline CV-8972, methods for preparing such compositions, and methods for treating various conditions in subjects using these compositions.

[0117] CV-8972 polymorph

[0118] As described in the following examples, CV-8972 crystals can exist in at least five polymorphic forms: form A, form B, form C, form D, and form E. Form A is a monohydrate, while forms B, D, and E are anhydrous. Form C was not obtained in a purified form, therefore its hydration state cannot be determined.

[0119] Crystals can form as salts of CV-8972. For example, crystals can form as hydrochloride salts of CV-8972.

[0120] Figure 1This is a space-filled 3D model of the crystal structure of the polymorph CV-8972, a form D polymorph. The polymorph is a trihydrochloride, and chloride ions are shown in green.

[0121] Figure 2 This is a space-filled 3D model of the crystal structure of the D polymorph of CV-8972 at room temperature. The polymorph is a trihydrochloride, and chloride ions are shown in green.

[0122] Figure 3 This is a space-filled 3D model of the crystal structure of polymorph A of CV-8972. The polymorph is a trihydrochloride, and chloride ions are shown in green.

[0123] Pharmaceutical Composition

[0124] This invention provides a pharmaceutical composition containing crystals of a polymorph of CV-8972. For example, the composition may contain CV-8972 crystals in form A, form B, form C, form D, or form E. The composition may be substantially free of one or more other polymorphs. For example, the composition may contain polymorph A and be substantially free of polymorphs of forms B, C, D, and E.

[0125] If a composition containing polymorphs of CV-8972 contains a major polymorph with a defined purity level, the composition may be substantially free of one or more other polymorphs of CV-8972. Purity may be expressed as the percentage of the amount of the major polymorph relative to the total weight of the two or more polymorphs of CV-8972.

[0126] In some embodiments, the total weight is the weight of all polymorphs of CV-8972 in the composition. For example, a composition containing polymorph A and substantially free of other polymorphs may contain form A at a defined weight percentage of all polymorphs of CV-8972 in the composition. For example, the composition may contain form A at least 95% by weight, at least 96% by weight, at least 97% by weight, at least 98% by weight, at least 99% by weight, at least 99.5% by weight, at least 99.6% by weight, at least 99.7% by weight, at least 99.8% by weight, or at least 99.9% by weight of all polymorphs of CV-8972 in the composition.

[0127] In some embodiments, the total weight is the weight of the selected polymorph of CV-8972 in the composition. For example, a composition containing polymorph A and substantially free of polymorph B may contain form A at a defined weight percentage of forms A and B. For example, the composition may contain form A at at least 95 wt%, at least 96 wt%, at least 97 wt%, at least 98 wt%, at least 99 wt%, at least 99.5 wt%, at least 99.6 wt%, at least 99.7 wt%, at least 99.8 wt%, or at least 99.9 wt% of forms A and B in the composition. Similarly, a composition containing polymorph A and substantially free of polymorphs B and C may contain form A at defined weight percentages of forms A, B, and C. For example, the composition may contain form A in at least 95% by weight, at least 96% by weight, at least 97% by weight, at least 98% by weight, at least 99% by weight, at least 99.5% by weight, at least 99.6% by weight, at least 99.7% by weight, at least 99.8% by weight, or at least 99.9% by weight of CV-8972 in the composition.

[0128] Alternatively or additionally, if a composition containing a polymorph of CV-8972 contains a minor polymorph below a defined level, the composition may be substantially free of one or more other polymorphic forms of CV-8972. The presence of a minor polymorph may be defined as a percentage of the total weight of one or more minor polymorphs of CV-8972.

[0129] In some embodiments, the total weight is the weight of all polymorphs of CV-8972 in the composition. For example, a composition containing polymorph A and substantially free of other polymorphs may contain all polymorphs other than form A at a defined weight percentage of all polymorphs of CV-8972 in the composition. For example, the composition may contain all polymorphs other than form A at less than 5% by weight, less than 4% by weight, less than 3% by weight, less than 2% by weight, less than 1% by weight, less than 0.5% by weight, less than 0.4% by weight, less than 0.3% by weight, less than 0.2% by weight, or less than 0.1% by weight of all polymorphs of CV-8972 in the composition.

[0130] In some embodiments, the total weight is the weight of the selected polymorph of CV-8972 in the composition. For example, a composition containing polymorph A and substantially free of polymorph B may contain form B at a defined weight percentage of forms A and B. For example, the composition may contain form B at less than 5 wt%, less than 4 wt%, less than 3 wt%, less than 2 wt%, less than 1 wt%, less than 0.5 wt%, less than 0.4 wt%, less than 0.3 wt%, less than 0.2 wt%, or less than 0.1 wt% of forms A and B in the composition. Similarly, a composition containing polymorph A and substantially free of polymorphs B and C may contain forms B and C at defined weight percentages of forms A, B, and C. For example, the composition may contain forms B and C in less than 5 wt%, less than 4 wt%, less than 3 wt%, less than 2 wt%, less than 1 wt%, less than 0.5 wt%, less than 0.4 wt%, less than 0.3 wt%, less than 0.2 wt%, or less than 0.1 wt% of forms A, B, and C of CV-8972 in the composition.

[0131] The composition may contain hydrochloride of the CV-8972 polymorph. The composition may contain CV-8972 and chloride ions in a defined stoichiometric ratio. The composition may contain CV-8972 and chloride ions in a 1:3 stoichiometric ratio.

[0132] The composition may contain a hydrated form of CV-8972. The composition may contain a monohydrate form of CV-8972, such as polymorph A. The composition may contain anhydrous forms of CV-8972, such as polymorphs B, D, or E.

[0133] This composition can be formulated for any route or mode of administration. It can be formulated for oral, skin, enteral, intra-arterial, intramuscular, intraocular, intravenous, nasal, oral, parenteral, pulmonary, rectal, subcutaneous, topical, or percutaneous application. It can also be formulated for administration by injection or with or on an implantable medical device (e.g., a stent, or a drug-eluting stent or balloon equivalent).

[0134] The composition can be formulated into a single unit dose. The composition can be formulated into separate doses.

[0135] The composition may contain a defined dose of CV-8972. This dose may contain approximately 10 mg to approximately 2000 mg, approximately 10 mg to approximately 1000 mg, approximately 10 mg to approximately 800 mg, approximately 10 mg to approximately 600 mg, approximately 10 mg to approximately 400 mg, approximately 10 mg to approximately 300 mg, approximately 10 mg to approximately 200 mg, approximately 25 mg to approximately 2000 mg, approximately 25 mg to approximately 1000 mg, approximately 25 mg to approximately 800 mg, approximately 25 mg to approximately 600 mg, approximately 25 mg to approximately 400 mg, approximately 25 mg to approximately 300 mg, approximately 25 mg to approximately 200 mg, approximately 50 mg to approximately 2000 mg, approximately 50 mg to approximately 1000 mg, approximately 50 mg to approximately 800 mg, approximately 50 mg to approximately 600 mg, approximately 50 mg to approximately 400 mg, approximately 50 mg to approximately 3 ...300 mg, approximately 50 mg to approximately 300 mg, approximately 50 mg to approximately 300 mg, approximately 50 mg to approximately 300 mg, approximately 50 mg to approximately 300 mg, approximately 50 mg to approximately 300 mg, approximately 50 mg to approximately 300 mg, approximately 50 mg to approximately 300 mg, approximately 50 mg to approximately 300 mg, approximately CV-8972 of mg to about 200 mg, about 100 mg to about 2000 mg, about 100 mg to about 1000 mg, about 100 mg to about 800 mg, about 100 mg to about 600 mg, about 100 mg to about 400 mg, about 100 mg to about 300 mg, about 100 mg to about 200 mg, about 200 mg to about 2000 mg, about 200 mg to about 1000 mg, about 200 mg to about 800 mg, about 200 mg to about 600 mg, about 200 mg to about 400 mg, about 200 mg to about 300 mg, about 300 mg to about 2000 mg, about 300 mg to about 1000 mg, about 300 mg to about 800 mg, about 300 mg to about 600 mg, or about 300 mg to about 400 mg. This dosage may contain approximately 10 mg, approximately 25 mg, approximately 50 mg, approximately 100 mg, approximately 200 mg, approximately 300 mg, or approximately 400 mg of CV-8972.

[0136] Pharmaceutical compositions containing the polymorph of CV-8972 can be in forms suitable for oral administration, such as tablets, sugar lozenges, tablets, instant dispersible powders, granules, or capsules. Compositions intended for oral use can be prepared according to any method known in the art for manufacturing pharmaceutical compositions, and such compositions may contain one or more agents selected from sweeteners, flavoring agents, coloring agents, and preservatives to provide a pharmaceutically palatable formulation. Tablets contain the polymorph mixed with non-toxic, pharmaceutically acceptable excipients. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrants such as corn starch or alginate; binding agents such as starch, gelatin, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. The preparation and administration of pharmaceutical compositions are discussed in U.S. Patent No. 6,214,841 and U.S. Patent Publication No. 2003 / 0232877, the contents of each of which are incorporated herein by reference. Formulations for oral use can also be present as hard gelatin capsules, wherein the compound is mixed with an inert solid diluent (such as calcium carbonate, calcium phosphate, or kaolin). This formulation allows for controlled release of the polymorph of CV-8972 in the gastrointestinal tract by encapsulating the polymorph in an enteric coating.

[0137] Dispersed powders or granules provide compounds that can be mixed with dispersants or wetting agents, suspending agents, and one or more preservatives. Examples of suitable dispersants or wetting agents and suspending agents include, for example, sweeteners, flavoring agents, and coloring agents.

[0138] The pharmaceutical composition may contain a mixture comprising an erodible polymer that promotes swelling of the mixture in an aqueous environment. An erodible polymer is any polymer that decomposes in vivo within a physiologically relevant timeframe. The erodible polymer may also have other characteristics that promote the gradual release of the polymorphic form of CV-8972 from the mixture. For example, but not limited to, the polymer may have one or more of the following: biocompatibility, i.e., harmless to living tissue; hydrophilicity; hygroscopicity; and a tendency to form a hydrogel.

[0139] Without being bound by theory, polymer-containing mixtures can facilitate gradual release through one or more mechanisms. For example, swelling of the mixture by absorbing water can promote the diffusion of polycrystalline forms of CV-8972 from the mixture. Polymer degradation can also allow the release of polycrystalline forms of CV-8972 from the mixture. Osmotic pressure generated by the high concentration gradient of compounds between the interior and exterior of the mixture can also contribute to the diffusion of polycrystalline forms of CV-8972 from the mixture.

[0140] For example, but not limited to, the polymer can be a cellulose derivative, a gelatin derivative, such as a cross-linked gelatin derivative or a polyester derivative.

[0141] Cellulose derivatives are linearly linked β(1→4) D-glucose units, comprising polymers with substitutions on one or more hydroxyl groups in each glucose unit. The substituents can be organic or inorganic and are typically linked via ester or ether bonds. Cellulose ester derivatives include carboxymethyl cellulose (CMC), such as sodium carboxymethyl cellulose, ethyl cellulose, ethyl hydroxyethyl cellulose, ethyl methyl cellulose, hydroxyethyl cellulose, hydroxyethyl methyl cellulose, hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose (HPMC), and methyl cellulose. Cellulose ether derivatives include cellulose acetate, cellulose acetate butyrate, cellulose acetate propionate, cellulose propionate, cellulose sulfate, cellulose triacetate, and nitrocellulose. The use of cellulose-based polymers to form biodegradable hydrogels is known in the art and is described, for example, in Sannino et al., Biodegradable Cellulose-based Hydrogels: Design and Applications, Materials, 2009, 2, 353-373; doi:10.3390 / ma2020353, the contents of which are incorporated herein by reference.

[0142] The mixture may contain multiple polymers or multiple polymeric forms of the same polymer. For example, HPMC polymeric forms may differ in various physical properties, including viscosity, degree of methoxy substitution, degree of hydroxypropoxy substitution, or average molecular weight.

[0143] The viscosity of the polymeric form of HMPC can be determined by testing under standard conditions, including the concentration of HMPC in the solution and the temperature of the solution. For example, but not limited to, the HMPC concentration can be 1%, 1.5%, 2%, 2.5%, or 3%. For example, but not limited to, the solution temperature can be 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, or 25°C.

[0144] Polymerized forms of cellulose derivatives, such as HPMC, can have defined viscosities. For example, but not limited to, for a 2% aqueous solution of polymeric HPMC at 20°C, the viscosity of the polymeric form can be about 2 cP to about 4 cP, about 4 cP to about 6 cP, about 5 cP to about 8 cP, about 12 cP to about 18 cP, about 40 cP to about 60 cP, about 80 cP to about 120 cP, about 300 cP to about 500 cP, about 1200 cP to about 2400 cP, about 2500 cP to about 5000 cP, about 9000 cP to about 18,000 cP, about 12,000 cP to about 24,000 cP, about 12,000 cP to about 24,000 cP, about 75,000 cP to about 150,000 cP, or at least about 2 cP. At least about 4 cP, at least about 5 cP, at least about 12 cP, at least about 40 cP, at least about 80 cP, at least about 300 cP, at least about 1200 cP, at least about 2500 cP, at least about 9000 cP, at least about 12,000 cP, at least about 12,000 cP, at least about 75,000 cP, less than about 4 cP, less than about 6 cP, less than about 8 cP, less than about 18 cP, less than about 60 cP, less than about 120 cP, less than about 500 cP, less than about 2400 cP, less than about 5000 cP, less than about 18,000 cP, less than about 24,000 cP, less than about 24,000 cP, or less than about 150,000 cP.

[0145] The polymeric forms of cellulose derivatives, such as HPMC, can differ in terms of the degree of substitution of their glucose units. The degree of substitution can be expressed as the weight percentage of substituents or the molar ratio of substituents to glucose units. For cellulose derivatives, such as HPMC, with two different substituents, the polymeric form can be described by the degree of substitution of each substituent.

[0146] Each polymeric form of HPMC can independently have a defined degree of methoxy substitution. For example, but not limited to, the degree of methoxy substitution can be about 19% to about 24%, about 22% to about 24%, about 27% to about 30%, about 27% to about 30%, or about 28% to about 32%.

[0147] Each polymeric form of HPMC can independently have a defined degree of hydroxypropoxy substitution. For example, but not limited to, the degree of hydroxypropoxy substitution can be about 4% to about 8%, about 7% to about 10%, about 7% to about 12%, about 8% to about 10%, about 8% to 11%, or about 9% to about 12%.

[0148] Each polymeric form of HPMC can independently have a defined average molecular weight. The average molecular weight can be about 10 kDa, about 13 kDa, about 20 kDa, about 26 kDa, about 41 kDa, about 63 kDa, about 86 kDa, about 110 kDa, about 120 kDa, about 140 kDa, about 180 kDa, or about 220 kDa.

[0149] When multiple forms of polymer, such as HPMC, are present, one or more polymeric forms may be present in defined amounts. For example, but not limited to, the polymer, such as HPMC, may contain about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, about 95 wt%, about 96 wt%, about 97 wt%, about 98 wt%, about 99 wt%, at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, at least 96 wt%, at least 97 wt%, at least 98 wt%, or at least 99 wt% of one polymeric form.

[0150] Pharmaceutical compositions may comprise sustained-release formulations containing one or more polymorphic forms of CV-8972. These formulations contain a mixture of one or more polymorphic forms of CV-8972 and one or more erosive polymers that promote swelling of the mixture in an aqueous environment. The hygroscopic and erosive properties of the polymers allow the mixture to form a hydrogel that slowly decomposes in the digestive tract of the subject. Therefore, the mixture promotes the stable release of the polymorphic form of CV-8972 and its metabolites into circulation.

[0151] The mixture may contain a limited amount of the polymorphic form of CV-8972. The mixture may contain at least 5 wt%, at least 10 wt%, at least 20 wt%, at least 30 wt%, at least 40 wt%, at least 50 wt%, at least 60 wt%, at least 70 wt%, or at least 80 wt% of the polymorphic form of CV-8972.

[0152] The mixture may contain CV-8972 in a defined weight ratio in the polycrystalline form and the polymer. For example, but not limited to, the mixture may contain components in weight ratios of about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 3:2, about 2:1, about 3:1, about 4:1, about 5:1, about 1:100 to about 100:1, about 1:100 to about 50:1, about 1:100 to about 20:1, about 1:100 to about 10:1, about 1:100 to about 5:1, about 1:100 to about 2:1, about 1:50 to about 100:1, about 1:50 to about 50:1, about 1:50 to about 20:1, about 1:50 to about 10:1, about 1:50 to about 5:1, about 1:50 to about 2:1, about 1:20 to about 100:1, about 1:20 to about 50:1, about 1:20 to about 20:1 CV-8972 and polymers in polymorphic forms, approximately 1:20 to approximately 10:1, approximately 1:20 to approximately 5:1, approximately 1:20 to approximately 2:1, approximately 1:10 to approximately 100:1, approximately 1:10 to approximately 50:1, approximately 1:10 to approximately 20:1, approximately 1:10 to approximately 10:1, approximately 1:10 to approximately 5:1, approximately 1:10 to approximately 2:1, approximately 1:5 to approximately 100:1, approximately 1:5 to approximately 50:1, approximately 1:5 to approximately 20:1, approximately 1:5 to approximately 10:1, approximately 1:5 to approximately 5:1, approximately 1:5 to approximately 2:1, approximately 1:3 to approximately 100:1, approximately 1:3 to approximately 50:1, approximately 1:3 to approximately 20:1, approximately 1:3 to approximately 10:1, approximately 1:3 to approximately 5:1, or approximately 1:3 to approximately 2:1.

[0153] Pharmaceutical compositions can be formulated for specific routes of administration. The drug can be formulated for oral, enteral, intravenous, or rectal administration.

[0154] The pharmaceutical composition can be formulated into unit doses containing a defined amount of CV-8972 in a polycrystalline form. These unit doses may contain approximately 5 mg, approximately 10 mg, approximately 20 mg, approximately 50 mg, approximately 100 mg, approximately 200 mg, approximately 500 mg, approximately 5 mg to approximately 10 mg, approximately 5 mg to approximately 20 mg, approximately 5 mg to approximately 50 mg, approximately 5 mg to approximately 100 mg, approximately 5 mg to approximately 200 mg, approximately 5 mg to approximately 500 mg, approximately 10 mg to approximately 20 mg, approximately 10 mg to approximately 50 mg, approximately 10 mg to approximately 10 ... CV-8972 in polycrystalline form, about 200 mg, about 10 mg to about 500 mg, about 20 mg to about 50 mg, about 20 mg to about 100 mg, about 20 mg to about 200 mg, about 20 mg to about 500 mg, about 50 mg to about 100 mg, about 50 mg to about 200 mg, about 50 mg to about 500 mg, about 100 mg to about 200 mg, about 100 mg to about 500 mg, or about 200 mg to about 500 mg.

[0155] The pharmaceutical composition can be formulated to produce defined values ​​of one or more parameters, as described below with respect to the method of the present invention. For example, but not limited to, the parameter can be C. max Application and Implementation C max The interval between, T 1 / 2 Or AUC.

[0156] The pharmaceutical compositions of the present invention may contain excipients. For example, but not limited to, the compositions may contain sweeteners, flavoring agents, coloring agents, or preservatives. These compositions may contain one or more of mannitol, starch, and magnesium stearate.

[0157] Provide subjects with polymorphs of CV-8972

[0158] This invention provides a method for treating a subject's condition by providing a polymorph of CV-8972. The polymorph may be of form A, form B, form C, form D, or form E. As described above, the polymorph of CV-8972 may be provided in a pharmaceutical composition. In some embodiments of this method, only the polymorph of form A is provided.

[0159] The polymorph of CV-8972 can be delivered via any suitable route or mode of administration. For example, but not limited to, the polymorph of CV-8972 can be delivered via oral, skin, enteral, intra-arterial, intramuscular, intraocular, intravenous, nasal, oral, parenteral, pulmonary, rectal, subcutaneous, topical, percutaneous, by injection, or through or onto an implantable medical device (e.g., a stent, or a drug-eluting stent or balloon equivalent).

[0160] The polymorph of CV-8972 can be provided according to the dosing regimen. The dosing regimen may include dosage, dosing frequency, or both.

[0161] Dosage can be delivered at any suitable interval. For example, but not limited to, it can be delivered once a day, twice a day, three times a day, four times a day, five times a day, six times a day, eight times a day, once every 48 hours, once every 36 hours, once every 24 hours, once every 12 hours, once every 8 hours, once every 6 hours, once every 4 hours, once every 3 hours, once every two days, once every three days, once every four days, once every five days, once a week, twice a week, three times a week, four times a week, or five times a week.

[0162] The dosage may contain a limited amount of CV-8972 that improves cardiac mitochondrial function, such as any dosage described above for pharmaceutical compositions containing polymorphs of CV-8972.

[0163] Dosage can be provided in a single dose, i.e., the dose can be provided as a single tablet, capsule, pill, etc. Alternatively, dosage can be provided in separate doses, i.e., the dose can be provided as multiple tablets, capsules, pills, etc.

[0164] Dosing may be administered for a specified period of time. For example, but not limited to, a dose may be provided for at least one week, at least two weeks, at least three weeks, at least four weeks, at least six weeks, at least eight weeks, at least ten weeks, at least twelve weeks, or longer.

[0165] The subject can be a person. The subject can be someone with cardiovascular disease, rheumatic disease, fibrosis, or cancer. The subject can be someone at risk of developing cardiovascular disease, rheumatic disease, fibrosis, or cancer. If a subject does not meet the established criteria for diagnosis of a condition but has one or more symptoms, markers, or other factors that indicate the subject may meet the diagnostic criteria for the condition in the future, then the subject may be at risk of developing the condition. The subject can be a child, newborn, infant, toddler, child, adolescent, young adult, young person, adult, or elderly subject. The subject can be in emergency care, intensive care, neonatal intensive care, pediatric intensive care, coronary care, cardiothoracic care, surgical intensive care, medical intensive care, long-term intensive care, operating room, ambulance, field hospital, or off-site environment.

[0166] Symptoms that can be treated with CV-8972 polymorph

[0167] This invention provides a method for treating a subject's condition by providing a polymorph of CV-8972. The condition can be any disease, symptom, or illness that provides therapeutic benefit by increasing mitochondrial energy production.

[0168] The symptoms can be cardiac symptoms. For example, but not limited to, these cardiac symptoms can include aneurysm, angina pectoris, atherosclerosis, cardiomyopathy, cerebrovascular disease, congenital heart disease, coronary artery disease (CAD), coronary heart disease, diabetic cardiomyopathy, heart attack, heart disease, heart failure, high blood pressure / hypertension, ischemic heart disease, pericardial disease, peripheral artery disease, refractory angina pectoris, rheumatic heart disease, stable angina pectoris, stroke, transient ischemic attack, unstable angina pectoris, or valvular heart disease.

[0169] Angina pectoris / angina is chest pain or pressure, usually caused by insufficient blood flow to the heart muscle. The pain or discomfort is located behind the sternum or on the left side and may radiate to the left arm, neck, jaw, or back. Several classifications of angina are known.

[0170] Stable angina, also known as exertional angina, is associated with myocardial ischemia. In stable angina, chest discomfort and related symptoms are often triggered by physical activity such as running or walking, but are mild or absent when the patient is at rest or has taken sublingual nitroglycerin. Symptoms usually lessen a few minutes after activity and recur upon resuming activity. Symptoms can also be triggered by cold weather, difficult-to-digest food, and emotional stress.

[0171] Unstable angina refers to changes or worsening of angina. Unstable angina has at least one of the following characteristics: (1) it occurs at rest or with light exercise and usually lasts for more than 10 minutes; (2) it is severe and new-onset, i.e., within the previous 4-6 weeks; and (3) it occurs in a progressive manner, i.e., it is significantly more severe, lasts longer or is more frequent than before.

[0172] Cardiac syndrome X, also known as microvascular angina, is angina-like chest pain against a background of normal epicardial coronary arteries during angiography. Its primary cause is unclear, but factors clearly associated with it include endothelial dysfunction and reduced flow in the heart's small resistance vessels. Microvascular angina may be part of the pathophysiology of ischemic heart disease.

[0173] Refractory angina is a chronic condition (lasting ≥3 months) in which angina (1) occurs in the context of coronary artery disease (CAD), (2) cannot be controlled by a combination of optimal drug therapy, angioplasty or bypass surgery, and (3) in which reversible myocardial ischemia has been clinically identified as the cause of the symptoms.

[0174] Providing polymorphs of CV-8972 can improve cardiac efficiency in subjects. Various definitions of cardiac efficiency exist in the medical literature. See, for example, Schipke, JD, Cardiac efficiency, *Basic Res. Cardiol.* 89:207-40 (1994); and Gibbs, CL and Barclay, CJ, Cardiac efficiency, *Cardiovasc. Res.* 30:627-634 (1995), which are incorporated herein by reference. One definition of cardiac mechanical efficiency is the ratio of external cardiac power to cardiac energy expenditure of the left ventricle. See Lopaschuk, G.D. et al., Myocardial Fatty Acid Metabolism in Health and Disease, *Physical Review* 90:207-258 (2010), which is incorporated herein by reference. Another definition is the ratio between stroke volume and oxygen consumption, which ranges from 20% to 25% in a normal human heart. (Visser, F., Measuring cardiac efficiency: is it useful?, Heart Metabolism 39:3-4 (2008), which is incorporated herein by reference.) Another definition is the ratio of stroke volume to mean arterial blood pressure. Any suitable definition of cardiac efficiency can be used to measure the effects of the compounds of this invention.

[0175] The polymorph of CV-8972 can be used to treat rheumatic diseases, conditions, or symptoms. As used herein, a rheumatic disease, condition, or symptom is any condition that affects or is associated with pain in or in one or more of these tissues, including joints, tendons, ligaments, bones, muscles, or connective tissue. Such a rheumatic disease, condition, or symptom may primarily affect joints, tendons, ligaments, bones, muscles, or connective tissue. Examples of such conditions include ankylosing spondylitis, autoimmune-associated lung disease, Behcet's disease, bursitis, chronic fatigue syndrome, dermatomyositis, fibromyalgia, gout, Gulf War syndrome, infectious arthritis, inflammatory arthritis, inflammatory eye disease, inflammatory myositis, juvenile idiopathic arthritis, lupus, myofascial pain syndrome, osteoarthritis, osteonecrosis of the jaw, osteoporosis, polymyalgia rheumatica, polymyositis, psoriatic arthritis, rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren's syndrome, tendinitis, and vasculitis.

[0176] This rheumatic disease, condition, or symptom may primarily affect the cardiovascular system and have secondary effects on joints, tendons, ligaments, bones, muscles, or connective tissue. For example, but not limited to, the symptom can be alcoholic cardiomyopathy, aneurysm, angina (including refractory angina and angina in the context of heart failure), atherosclerosis, cardiac fibrosis, cardiomyopathy, cerebrovascular disease, claudication (e.g., peripheral claudication), congenital heart disease, coronary artery disease, coronary artery disease, cyanotic heart disease, diabetic cardiomyopathy, dilated cardiomyopathy, heart attack, heart failure, hypertension, hypertrophic cardiomyopathy, ischemic cardiomyopathy, ischemic heart disease, left ventricular dysfunction, pericardial disease, peripheral artery disease, rheumatic heart disease, stroke, transient ischemic attack, or valvular heart disease.

[0177] This rheumatic disease, condition, or symptom can be a rare muscle disorder. For example, but not limited to, it may be CAV3-associated distal myopathy, Duchenne Muscular Dystrophy, hypertrophic cardiomyopathy, isolated hyperCKemia, limb-girdle muscular dystrophy, myomyopathy, myositis, or rhabdomyomyopathy. This rare muscle disorder may be associated with mutations in BICD2, CAV3, or DMD.

[0178] The rheumatic disease, condition, or symptom can be glycogen storage disorder. For example, but not limited to, it can be aldolase A deficiency, Andersen disease, Cori's disease, Fanconi-Bickel syndrome, Her's disease, Laforadisease, McArdle disease, Pompe's disease, Tarui's disease, or von Gierke's disease. This glycogen storage disorder may be related to enzyme or protein deficiencies, such as…

[0179] The enzymes involved include α-glucosidase, aldolase A, β-enolase, glucose transporter, glucose-6-phosphatase, glycogen branching enzyme, glycogen debranching enzyme, glycogen synthase, glycogen protein-1, liver glycogen phosphorylase, muscle glycogen phosphorylase, muscle lactate dehydrogenase, muscle phosphofructokinase, muscle phosphoglycerate mutase, phosphoglycerate mutase, or phosphorylase kinase. This glycogen storage disorder may be associated with mutations in genes such as AGL, ALDOA, ENO3, G6PC, GAA, GBE1, GLUT2, GYG1, GYS2, LDHA, PGAM2, PGAM2, PHKA1, PHKA2, PHKB, PHKG2, PKFM, PYGL, PYGM, or SLC37A4.

[0180] The rheumatic disease, condition, or symptom may be another condition affecting joints, tendons, ligaments, bones, muscles, or connective tissue, such as acute kidney injury, cachexia, chemotherapy-induced nephropathy, contrast-induced nephropathy, imbalance, weakness, pulmonary hypertension, pulmonary fibrosis, sarcopenia, tinnitus, or vertigo.

[0181] Polymorphs of CV-8972 can be used to treat fibrosis or diseases, conditions, or symptoms associated with fibrosis. Specifically, these methods can be used to treat diseases, conditions, or symptoms related to fibrosis in an organ or tissue and a reduction in the energy production of that organ or tissue. Fibrosis can affect any organ or tissue, such as the heart, lungs, liver, brain, cardiovascular system, joints, gastrointestinal system, limbs, fingers, skin, bone marrow, or penis.

[0182] The fibrosis may be associated with another condition, for example, it may be secondary to another condition, or it may cause another condition. For example, but not limited to, the fibrosis may include or be related to the following: adhesive capsulitis, aneurysm, angina pectoris, arteriosclerosis, joint fibrosis, atherosclerosis, atrial fibrosis, cardiomyopathy, cerebrovascular disease, cirrhosis, congenital heart disease, coronary artery disease, coronary heart disease, Crohn's disease, cystic fibrosis, diabetic cardiomyopathy, Dupuytren's contracture, endocardial fibrosis, glial scars, heart attack, heart failure, high blood pressure / hypertension, idiopathic pulmonary fibrosis, ischemic heart disease, keloids, mediastinal fibrosis, myelofibrosis, renal systemic fibrosis, old myocardial infarction, pericardial disease, peripheral artery disease, Peroni's disease, progressive massive fibrosis, pulmonary fibrosis, radiation-induced lung injury, retroperitoneal fibrosis, rheumatic heart disease, scleroderma, stroke, systemic sclerosis, transient ischemic attack, or valvular heart disease.

[0183] Polymorphs of CV-8972 can be used to treat cancer. For example, but not limited to, the cancer can be bladder cancer, brain cancer, breast cancer, carcinoma, cervical cancer, colon cancer, colorectal cancer, stomach cancer, glioblastoma, glioma, head and neck cancer, kidney cancer, leukemia, liposarcoma, liver cancer, lung cancer, lymphoma, medulloblastoma, melanoma, muscle cancer, neuroblastoma, oligodendroglioma, oligodendroglioma, osteosarcoma, ovarian cancer, pancreatic cancer, paraganglioma, prostate cancer, sarcoma, or thyroid cancer.

[0184] Synthesis of CV-8972

[0185] This invention also provides a synthetic scheme for CV-8972, wherein the free base form of CV-8814, which forms the product in a reductive amination reaction, can be directly used as a substrate in an esterification reaction. This invention is partly based on the identification of conditions that improve the stability of the free base of CV-8814 and allow the free base form to crystallize. Therefore, the scheme provided herein eliminates the need to convert CV-8814 from its free base form to an HCl salt and then back to the free base form. Thus, this invention provides a simpler, faster, and higher-yield method for preparing CV-8972.

[0186] This invention provides a method for preparing compounds of formula (X):

[0187]

[0188] This preparation is carried out by performing the following steps:

[0189] React 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol to produce the free base form of compound (IX):

[0190] as well as

[0191] The free base form of compound (IX) is reacted with nicotinic acid to produce compound (X).

[0192] This method does not include the salt form that produces the compound of formula (IX).

[0193] 2,3,4-Trimethoxybenzaldehyde has the following structure:

[0194]

[0195] 2-(piperazin-1-yl)ethane-1-ol has the following structure:

[0196]

[0197] Nicotinic acid has the following structure:

[0198]

[0199] The step of reacting 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol-1-ol may include one or more solvents, catalysts, or other chemicals. The step of reacting 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol-1-ol may include one or more of sodium triacetoxyborohydride, acetic acid, and 2-methyltetrahydrofuran.

[0200] The reaction of 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol-1-ol can be carried out at a defined temperature. The reaction can be carried out at approximately 10°C to approximately 30°C, approximately 15°C to approximately 30°C, approximately 20°C to approximately 30°C, approximately 25°C to approximately 30°C, approximately 10°C to approximately 25°C, approximately 15°C to approximately 25°C, approximately 20°C to approximately 25°C, approximately 10°C to approximately 20°C, or approximately 15°C to approximately 20°C.

[0201] The step of reacting 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol-1-ol may not involve a specific solvent, catalyst, or other chemical. The step of reacting 2,3,4-trimethoxybenzaldehyde with 2-(piperazin-1-yl)ethanol-1-ol may not involve dichloromethane.

[0202] The step of reacting the free base form of compound (IX) with nicotinic acid can produce the free base form of compound (X).

[0203] The step of reacting the free base form of compound (IX) with nicotinic acid may include one or more solvents, catalysts, or other chemicals. The step of reacting the free base form of compound (IX) with nicotinic acid may include one or more of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, 4-(dimethylamino)pyridine, and dichloromethane.

[0204] The step of reacting the free base form of compound (IX) with nicotinic acid can be carried out at a defined temperature. The step of reacting the free base form of compound (IX) with nicotinic acid can be carried out at about 15°C to about 30°C, about 20°C to about 30°C, about 25°C to about 30°C, about 15°C to about 25°C, about 20°C to about 25°C, or about 15°C to about 20°C.

[0205] The method may include converting the free base form of compound (X) into a salt form of compound (X). The salt form of compound (X) may be an HCl salt. The salt form of compound (X) may be a monohydrate.

[0206] The step of converting the free base form of compound (X) to the salt form of compound (X) may include one or more solvents, catalysts, or other chemicals. The step of converting the free base form of compound (X) to the salt form of compound (X) may include one or more of HCl and methyl ethyl ketone.

[0207] The step of converting the free base form of compound (X) to the salt form of compound (X) can be carried out at a defined temperature. The step of converting the free base form of compound (X) to the salt form of compound (X) can be carried out at approximately 40°C to approximately 60°C, approximately 45°C to approximately 60°C, approximately 50°C to approximately 60°C, approximately 55°C to approximately 60°C, approximately 40°C to approximately 55°C, approximately 45°C to approximately 55°C, approximately 50°C to approximately 55°C, approximately 40°C to approximately 50°C, approximately 40°C to approximately 50°C, approximately 40°C, approximately 45°C, approximately 50°C, approximately 55°C, or approximately 60°C.

[0208] The compound of formula (X) can exist in at least five crystalline forms: form A, form B, form C, form D, and form E. Form A is a monohydrate, and forms B, D, and E are anhydrous. The method may include converting the compound of formula (X) from a first crystalline form to a second crystalline form. Each of the first and second crystalline forms can independently be form A, form B, form C, form D, or form E. The method may include one or more of the following conversions of the compound of formula (X): conversion from an anhydrous form to a hydrated form; conversion from a hydrated form to an anhydrous form; conversion from one anhydrous form to another anhydrous form; and conversion from one hydrated form to another hydrated form.

[0209] The step of converting the salt form of compound (X) from the first crystalline form to the second crystalline form may include one or more of the following: changing the solvent of the salt form of compound (X) and incubating the salt form of compound (X) at about 60°C.

[0210] This method can be carried out without the use of one or more solvents, catalysts, or other chemicals. This method can also be carried out without the use of one or more of dioxane, ethyl acetate, or potassium carbonate.

[0211] The method may include purifying the free base form of the compound of formula (IX). The method may also include crystallizing the free base form of the compound of formula (IX).

[0212] Example

[0213] Example 1

[0214] Overview

[0215] A comprehensive polymorph screening was conducted on CV-8972, which has the structure of formula (X). The starting material CV-8972 was characterized by X-ray powder diffraction (XRPD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), dynamic vapor adsorption (DVS), and polarized light microscopy (PLM). The data indicate that the material is inherently crystalline and possesses an XRPD pattern similar to that of form A. Starting with form A, a polymorph / single-crystal screening experiment was established under 34 conditions using vapor diffusion, slow evaporation, and cooling crystallization methods. Five distinct XRPD patterns were observed, encompassing forms A, B, A+C, D, and E. Form A is a monohydrate form, as confirmed by its single-crystal structure. Form D is anhydrous, and this was also confirmed by its single-crystal structure. Form E is an anhydrous form produced by dehydration of form A at approximately 90 °C. Form B is a known anhydrous form from another study. During the study, form C was not obtained in its pure form, but rather as a mixture of forms A and C. Water activity analysis indicated that form E converted to form A under all test conditions. Furthermore, form E showed partial conversion to form A when exposed to ambient temperature and humidity. Further, results from slurry competition between the two anhydrous forms D and E also indicated that both forms converted to form A during the experiment. These results suggest that form A is the most stable form at ambient temperature and humidity.

[0216] Representation of form A

[0217] The starting material of CV-8972 was characterized using X-ray powder diffraction (XRPD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and polarizing microscopy (PLM).

[0218] Figure 4 This is the XRPD diffraction pattern of the CV-8972 starting material. The XRPD results indicate a high crystallinity of the starting material. Comparison of the XRPD of the starting material with previously known polymorphs indicates that it is of form A.

[0219] Figure 5 The TGA and DSC thermograms of the CV-8972 starting material are shown. The TGA thermogram is shown in green, and the DSC thermogram is shown in blue. As the TGA and DSC data indicate, a weight loss of approximately 3.46% was observed at up to 150 °C before decomposition. The DSC shows a small endothermic reaction at 85.3 °C (peak), and a possible melting endothermic reaction at 214.6 °C (start), followed by decomposition, and a melting point of 131.7 °C (peak) is observed.

[0220] Figure 6 XRPD diffraction patterns of CV-8972 in various forms are shown. The starting material of CV-8972 is shown in blue; CV-8972 after incubation at 90°C for 8 hours is shown in red; and CV-8972 after incubation at 65°C in vacuum for 2 hours is shown in purple. To observe whether the small DSC endothermic reaction at 85.3°C corresponds to a polycrystalline phase transformation or dehydration, form A was stored in an oven at 90°C for 8 hours and then subjected to XRPD. The data indicate that form A transforms into form E.

[0221] Figure 7 This is a polarization micrograph of the starting material CV-8972. PLM reveals a very flat, mica-like morphology of the crystals.

[0222] Figure 8 This is a dynamic vapor adsorption isotherm plot. Cyclic adsorption is shown in red; desorption in cycle 1 is shown in blue; and adsorption in cycle 2 is shown in green. The DVS results show that CV-8972 has a water absorption rate of <0.2% at 25°C and 80% relative humidity (RH), indicating that the starting material is non-hygroscopic. However, the mass change increases sharply above 80% RH, indicating that deliquescence may be present.

[0223] Figure 9 XRPD diffraction patterns of CV-8972 before and after dynamic vapor adsorption (DVS) are shown. Data before DVS are shown in red; and data after DVS are shown in blue. The XRPD of the sample after DVS indicates a weak crystalline peak, but is generally similar to that of the starting material.

[0224] Figure 10 XRPD diffraction patterns of the dehydrated and rehydrated forms of CV-8972 are shown. Data from the starting material are shown in blue; data after incubation in a vacuum oven for 2 hours are shown in red; and data from the heated material exposed to ambient relative humidity are shown in green. To monitor form A in its dehydrated state, it was placed in a vacuum oven at 65°C for 2 hours, followed by XRPD analysis. The XRPD results indicate that this process produced a new anhydrous form of the material, designated as form E. Upon exposure to ambient RH, rehydration of form E caused a partial conversion to form A.

[0225] X-ray powder diffraction data have been difficult to interpret due to extremely preferred orientations, resulting in large variations in peak intensity from one sample preparation to the next. To minimize this effect, single-crystal X-ray diffraction was used to obtain the crystal structure, and the X-ray powder diffraction that should be observed in an ideal sample without preferred orientations was calculated.

[0226] Polymorph / Single Crystal Screening

[0227] Starting with Form A, a polymorph screening experiment was established under 34 conditions using slurry conversion, liquid-gas diffusion, slow evaporation, and slow cooling methods. The approximate solubility of the starting materials was determined at room temperature (RT). Approximately 2 mg of the sample was precisely weighed and added to a 3-mL glass vial. Solvent was then added incrementally (50 / 50 / 200 / 700 μL) to the vial until the solid dissolved or a total volume of 1 mL was reached. Table 1 shows the solubility of the starting materials in various solvents.

[0228] Table 1: Approximate solubility of starting material (6010242-01-A) at RT

[0229] Experiment ID Solvent (v:v) Solubility (mg / mL) 6010242-02-A1 n-Heptane S<1.9 6010242-02-A2 ACN S<1.5 6010242-02-A3 MIBK S<2.0 6010242-02-A4 EtOAc S<1.8 6010242-02-A5 THF S<2.2 6010242-02-A6 EtOH S<1.7 6010242-02-A7 acetone S<2.0 6010242-02-A8 MEK S<2.0 6010242-02-A9 IPA S<2.3 6010242-02-A10 <![CDATA[CHCI3]]> S<1.8 6010242-02-A11 IPAc S<1.7 6010242-02-A12 1,4-Dioxane S<1.7 6010242-02-A13 CPME S<1.5 6010242-02-A14 DCM S<2.2 6010242-02-A15 Toluene S<2.3 6010242-02-A16 DMSO 5.0<S<15 6010242-02-A17 DMF 1.8<S<6.0 6010242-02-A18 NMP 6.7<S<20 6010242-02-A19 <![CDATA[H2O]]> S>44.0 6010242-02-A20 MeOH S>46.0

[0230] Solubility analysis results were used to guide solvent selection in polymorph screening. Polymorph screening experiments were conducted using different crystallization or solid-state transformation methods. Table 2 summarizes the polymorph screening experiments.

[0231] Table 2:

[0232] method Experiment number Crystal type Liquid vapor diffusion 24 Forms A, B, C, D, and E Slow evaporation 2 gel Slow cooling 2 No sediment Slurry conversion 6 Form A total 34 Forms A, B, C, D, and E

[0233] Figure 11 XRPD diffraction patterns of various polymorphs of CV-8972 are shown. Form A is shown in blue; Form B in green; a mixture of Forms A and C in dark blue; Form D in orange; and Form E in purple.

[0234] Table 3 summarizes the various crystal forms of CV-8972.

[0235] Table 3:

[0236]

[0237] These forms of single-crystal structures are available and provided as a separate report.

[0238] # Information obtained from separate reports

[0239] Liquid-gas diffusion experiments were conducted under different solvent conditions. Approximately 15–25 mg of the starting material was dissolved in a suitable solvent to obtain a clear solution in a 3–mL vial. This solution was then transferred to a 20–mL vial containing 3 mL of a volatile solvent. The 20–mL vial was capped and kept at room temperature, which allowed sufficient time for the organic vapors to interact with the solution. The precipitate was separated for XRPD analysis. Table 4 summarizes the results of the liquid-gas diffusion experiments.

[0240] Table 4:

[0241]

[0242]

[0243] Slow evaporation experiments were conducted under various conditions. In short, saturated solutions of the starting material prepared in different solvents were added to HPLC vials. The visually clear solution was then purified using a syringe with 5–10 pinholes. The mixture was covered and subjected to evaporation at RT. The solids were then separated for XRPD analysis. Table 5 summarizes the results of the slow evaporation experiments.

[0244] Table 5:

[0245] Experiment ID Solvent (v:v) solid form 6010242-06-A1 MeOH White solid, poorly crystallizing. 6010242-06-A2 <![CDATA[H2O]]> gel

[0246] Slow cooling experiments were conducted in two different solvent systems. Approximately 10–15 mg of the starting material was suspended at RT in a suitable solvent in a 2–mL glass vial. The suspension was then heated to 50 °C, equilibrated for approximately two hours, and filtered using a nylon membrane (0.22 μm pore size). Each filtrate was slowly cooled to 5 °C at a rate of 0.1 °C / min. Table 6 summarizes the results of the slow cooling experiments.

[0247] Table 6:

[0248] Experiment ID Solvent (v:v) solid form 6010242-05-A1 DMSO No sediment 6010242-05-A2 NMP No sediment

[0249] Slurry conversion experiments were conducted at RT in different solvent systems. Approximately 20 mg of the starting material was suspended in 0.1 mL of solvent in an HPLC vial. After magnetic stirring of the suspension at RT for 48 hours, the remaining solids were separated for XRPD analysis. Table 7 summarizes the results of the slurry conversion experiments.

[0250] Table 7:

[0251] Experiment ID Solvent (v:v) solid form 6010242-15-A1 acetone Form A 6010242-15-A2 <![CDATA[Acetone / H2O(a w = 0.2, 941 / 59)]]> Form A 6010242-15-A3 <![CDATA[Acetone / H2O(a w = 0.4, 857 / 143)]]> Form A 6010242-15-A4 <![CDATA[Acetone / H2O(a w = 0.6, 726 / 274)]]> Form A 6010242-15-A5 <![CDATA[Acetone / H2O(a w = 0.8, 492 / 508)]]> Form A 6010242-15-A6 <![CDATA[H2O]]> Form A 6010242-15-A7 Forms D+E in acetone Form A

[0252] in conclusion

[0253] Form A was successfully characterized to understand its form behavior. A comprehensive polymorph screening was conducted under 34 different conditions. Five polymorphs of CV-8972 were identified during the screening, including mixtures of forms A, B, A+C, D, and E. Forms D and E are anhydrous, form A is a monohydrate, and form B is a stoichiometric hydrate. The phase origin of form C is unknown because it is not obtained in its pure form; it always crystallizes as a mixture of forms A. Based on the polymorph screening, it is clear that CV-8972 has a tendency to form multiple polymorphs. The current study concludes that form A is the optimal form for developing CV-8972 and is a stable monohydrate form, and is the most stable form under ambient temperature and humidity conditions.

[0254] Instruments and methods

[0255] Use starting material analysis form A to screen for other polymorphs.

[0256] XRPD was performed using Panalytical X'Pert3 powder on a Si zero-background holder. The 2θ position was calibrated according to the Panalytical Si reference standard disk. The instrument parameters used for XRPD are listed in Table 8.

[0257] Table 8:

[0258]

[0259] TGA data were collected using a TA Discovery 550TGA from TA Instruments. DSC was performed using a TA Q2000 DSC from TA Instruments. The DSC was calibrated with an indium reference standard, and the TGA was calibrated with a nickel reference standard. Detailed parameters for the TGA and DSC are listed in Table 9.

[0260] Table 9:

[0261]

[0262]

[0263] Polarizing microscopy (PLM) images were captured at room temperature using a Nikon DS-Fi2 upright microscope. Low-viscosity microscope impregnation oil was used. Dispersed powder crystals.

[0264] Example 2

[0265] Overview

[0266] To determine the crystal structure of CV-8972, a monohydrate crystal was grown, and whole-crystal X-ray diffraction (SCXRD) data were collected using a suitable single crystal at 199 K. A crystal structure with an R1 value of 0.0303 (I>2σ(I)) was obtained. The structure indicates that the crystal form is a monohydrate triHCl salt.

[0267] Crystal growth and SCXRD preparation

[0268] C is obtained through slow cooling. 22 H 34 Single crystals of Cl3N3O6 (CV-8972): 163.2 mg of starting material was weighed into a 2-mL glass vial, and 0.100 mL of water was added to dissolve the solid at 50 °C. The solution was then slowly cooled to 10 °C within 12 hours before collection.

[0269] Figure 12 It is C 22 H 34 A series of PLM images of single crystals of Cl3N3O6 (CV-8972). Scale bar indicates 100 μm.

[0270] Figure 13 PLM images of the crystal used for a single-crystal diffractometer are shown. The scale bar indicates 100 μm. A coarse needle was removed and trimmed into a uniform block measuring 200 × 160 × 100 μm. This sample was mounted in a low-viscosity refrigeration oil (MiTeGenLV CryoOil). TM 100mm MiTeGen MicroLoop TM superior.

[0271] Figure 14 An image of a crystal mounted on a 100-micron Mitegen ring is shown.

[0272] Single crystal structure determination

[0273] A total of 9576 frames were collected using Bruker Apex3 v2018-7.2. The total exposure time was 18 hours (adjusted for exposure time based on 2θ). The frames were integrated with the Bruker SAINT software package using a narrow frame algorithm. Integrating the data using orthogonal cells yielded 157237 reflections, with a maximum θ angle of 81.01° (resolution). Of these, 5641 were independent (average redundancy 27.874, integrity = 99.8%, R...). int =4.41%, R sig =1.34%), and 5388 times (95.51%) greater than 2σ(F 2 Final cell constant and This is based on a refinement of the XYZ centroids of 1406 reflections above 20σ(I), where 11.75° < 2θ < 100.6°. Absorption effect correction was performed on the data using the SADABS method. The ratio of minimum to maximum apparent transmittance is 0.788. The calculated minimum and maximum transmittance coefficients (based on crystal size) are 0.5340 and 0.7160, respectively.

[0274] Using Olex2 in conjunction with the SHELXTL software package, the structure was solved and optimized using the orthogonal space group Pbca, with the specific unit C. 22 H 34 Cl3N3O6, Z=8. An asymmetric unit contains a complete API molecule. For observational data, for F with 330 variables (0 constraints) 2 The final anisotropic full-matrix least-squares refinement converges to R1 = 3.03%, and for all data, wR2 = 7.98%. The goodness of fit is 1.041. The maximum peak value in the final differential electron density synthesis is... ( From Cl1), and the largest hole is ( (From Cl1). The positions and thermal ellipsoids of most hydrogen atoms are considered as riding models (using AFIX 23, AFIX 43, and AFIX 137). However, key hydrogen atoms involved in hydrogen bonding and salt formation are freely refined without any restrictions. Based on the final model, the calculated density is 1.400 g / cm³. 3 And F(000), 2288e-. Table 10 summarizes C 22 H 34 Crystallographic parameters of Cl3N3O6 crystal.

[0275] Table 10:

[0276]

[0277]

[0278] Figure 15 It is C 22 H 34 Ortep diagram of the asymmetric unit cell of Cl3N3O6 crystal. 22 H 34 Ortep diagrams of the asymmetric units of Cl3N3O6 crystals indicate that the API is a monohydrate triHCl salt, as observed in the ratio 1:3:1 (API:HCl:H2O).

[0279] Figure 16 C is shown 22 H 34 A unit cell of Cl3N3O6 crystal.

[0280] Figure 17 It is C 22 H 34 A diagram of the hydrogen bond network and counter ion pairs in Cl3N3O6 crystals. The diagram shows that three hydrochloride molecules are deprotonated, while three nitrogen molecules are protonated. Water molecules act as hydrogen bond donors bridging the two chloride anions. Table 11 summarizes the C... 22 H 34 Crystallographic measurements of hydrogen bonds and counter ion pairs in Cl3N3O6 crystal.

[0281] Table 11:

[0282]

[0283] Equivalent atoms are generated using symmetry transformations: #1: 2-X, 0.5+Y, 0.5-Z; #2: 0.5+X, +Y, 0.5-Z; #3: 1+X, +Y, +Z; #4: 1.5-X, 0.5+Y, +Z

[0284] Local inspection of the final CIF file using Olex2 and Platon revealed only one Level C alert (missing three reflections) and eight Level G alerts. To prevent this issue, an extensive data collection strategy was implemented, resulting in a dataset with 99.8% integrity and 27.87% redundancy.

[0285] Figure 18 C is shown 22 H 34 Calculated and measured XRPD patterns of Cl3N3O6 crystals. The calculated XRPD diffraction pattern is shown in red; and the measured XRPD diffraction pattern is shown in blue. Powder X-ray diffraction for this batch was obtained and compared with the calculated pattern based on this crystal structure using Mercury. The experimental peak positions and intensities are in excellent agreement with the calculated pattern.

[0286] Instruments and methods

[0287] X-ray intensity data were measured at 199.0 K on a Bruker Venture X-ray diffractometer (controlled by an Oxford Cryostream 800). Monochromatic Cu Kα radiation was used with an Incoatec micro-focusing source (1 μS 3.0). A voltage of 50 kV and a current of 1.1 mA were used as the X-ray source. Intensity data were collected using a Photon II detector.

[0288] Polarizing microscopy images were captured using a Nikon DS-Fi2 vertical microscope at room temperature.

[0289] XRPD was performed using Panalytical X'Pert3 powder on a Si zero-background holder. The 2θ position was calibrated according to the Panalytical Si reference standard disk.

[0290] Example 3

[0291] Overview

[0292] To determine the crystal structure of CV-8972, a single anhydrous crystal of CV-8972 was grown, and full SCXRD data were collected using a suitable single crystal at 102 K. A crystal structure with an R1 value of 0.0328 (I>2σ(I)) was obtained. The structure indicates that the crystal form is an anhydrous triHCl salt.

[0293] Crystal growth and SCXRD preparation

[0294] Anhydrous triHCl salt single crystals of CV-8972 were obtained by liquid-gas diffusion of MTBE in MeOH solution. In short, a saturated solution of CV-8972 in MeOH was obtained at RT and filled into 2-mL glass vials, which were then stored in a larger 20mL vial containing 2mL of MTBE. The vial was removed when white crystalline material was observed.

[0295] Figure 19 A PLM image of a single crystal-free body from recrystallized CV-8972 is shown.

[0296] Figure 20 This is an image of a single anhydrous crystal from recrystallization site CV-8972, mounted on a glass fiber tip. The colorless crystal was subsequently set on the SCXRD instrument.

[0297] Single crystal structure determination

[0298] A colorless crystal was mounted on top of a glass fiber. X-ray intensity data were measured at 102 K using an ω / π scan technique on a Bruker D8 Quest PHOTON 100 CMOS X-ray diffractometer system with an Incoatec Microfocus Source (1 μS) monochromatic Mo Kα radiation. (Sealed tube). Data was collected in 1660 frames, with an exposure time of 10 seconds. Crystallographic data: C 22 H 32 O5N3Cl3: α=78.252(2)o, β=82.823(2)o, γ=82.476(2)o, Z=2, FW=524.85, μ=0.403mm-1, d=1.389g / cm 3 F(000) = 552.

[0299] Table 12 provides the crystal data and structural refinement for j1_a.

[0300] Table 12:

[0301]

[0302]

[0303] Table 13 provides the atomic coordinates of j1_a (x 10). 4 and equivalent isotropic displacement parameters U(eq) is defined as one-third of the trace of the orthogonalized Uij tensor.

[0304] Table 13:

[0305]

[0306]

[0307] Table 14 provides the bond length of j1_a.

[0308] Table 14:

[0309]

[0310]

[0311] Table 15 provides the angle [°] for j1_a.

[0312] Table 15:

[0313]

[0314]

[0315] At a maximum θ angle of 32.02° (resolution) Of the 8732 unique reflections of the Mo Kα radiation, 7553 were observed (I>2σ(Ι)). The linear absorption coefficient of the Mo Kα radiation is 0.403 mm- 1 The data was integrated with the manufacturer's SAINT software, and the absorption effect was corrected using the multiple scan method (SADABS).

[0316] Subsequent solving and optimization were performed using the SHELXTL-2014 solution package running on a Pentium computer. The structure was solved directly using the SHELXTL-2014 package. The non-hydrogen atom scattering factors were taken from a literature table. The positions of the non-hydrogen atoms were determined by successive Fourier map calculations. In the final refinement cycle of each refinement, all non-hydrogen atoms were refined in anisotropic shift parameters. Except for H(1), H(2), and H(3) on N(1), N(2), and N(3) in the molecule, which were located according to the Fourier map and refined under appropriate constraints, the positions of the remaining hydrogen atoms were calculated and allowed to ride on the carbons they are attached to, assuming a CH bond length of 1. (For CH2 groups, m = 0.990; for CH3 groups, m = 0.980; for Ph-H groups, m = 0.950). The hydrogen atom temperature factor is fixed at n (for CH2 and Ph-H groups, n = 1.2; for CH3, n = 1.5) times the isotropic temperature factor of the C atom it is attached to. The compound is triclinic, space group P-1 (number 2). The final residual values ​​based on 310 variable parameters and 7553 observed reflections (I > 2σ(Ι)) are R1 = 0.0328, wR2 = 0.0926, and the final residual values ​​for all unique reflections are R1 = 0.0408, wR2 = 0.0975. The goodness-of-fit index for all data is 1.014. The final difference plot ranges from 0.549 to The peak value has no chemical significance. Efforts have been made to resolve as many alerts generated by the CheckCIF procedure as possible. The current highest alert level is G.

[0317] Figure 21 It is C 22 H 32 A thermal ellipsoid diagram of the asymmetric unit cell of Cl3N3O5 crystal. This diagram shows that this form is an anhydrous triHCl salt.

[0318] Figure 22 C is shown 22 H 32 A unit cell of Cl3N3O5 crystal.

[0319] Figure 23 It is C 22 H 32 Diagram of hydrogen bond network and counter ion pairs in Cl3N3O5 crystal.

[0320] Figure 24 C is shown 22 H 34Calculated and measured XRPD patterns of Cl3N3O6 crystals. The calculated XRPD diffraction pattern is shown in red; and the measured XRPD diffraction pattern is shown in green.

[0321] The compound crystallizes in the triclinic system, space group P-1 (number 2). The asymmetric units contain molecules in the form of cationic / anionic salts (anhydrous triHCl salts), with the molecular formula C2. 22 H 32 O5N3Cl3. Intramolecular H bonds may exist between N(1)-H(1)...Cl(3) (where the distance is 2.9634(10)), N(2)-H(2)...Cl(2) (where the distance is 2.9822(9)), and N(3)-H(3)...Cl(1) (where the distance is 3.0120(9)). The exported results were structurally solved, optimized, and calculated using the SHELXTL-2014 computer software package. The neutral atom scattering factor is the Cromer and Waber neutral atom scattering factor, and the real and virtual anomalous dispersion corrections are Cromer's real and virtual anomalous dispersion corrections.

[0322] Example 4

[0323] Introduction

[0324] CV-8972 was synthesized according to scheme 1.

[0325] Option 1:

[0326]

[0327]

[0328] Step 1 involves reductive amination using 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethane-1-ol as starting materials, with sodium triacetoxyborohydride (STAB) as a reducing agent, in the presence of catalytic acetic acid (AcOH) and 2-methyltetrahydrofuran (2-MeTHF) as solvents. After the reaction is complete, an aqueous post-treatment is performed, in which the solvent is exchanged for MTBE and recrystallized from MTBE / n-heptane to form the intermediate CV-8814 free base (CV8814 free base).

[0329] In step 2, CV-8814 free base (CV8814 free base) is acid-coupled with nicotinic acid in dichloromethane (DCM) solvent in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and catalytic 4-(dimethylamino)pyridine (DMAP). After aqueous post-treatment, CV8972 free base is formed. The solvent is exchanged for 2-butanone (MEK), and then added to MEK containing concentrated HCl to form CV8972 monohydrate intermediate.

[0330] The final step, step 3, involves a form conversion in a mixture of water, methanol, and MEK at 60℃±5℃, followed by precipitation with the addition of MEK, and XRPD analysis to obtain the desired final product CV-8972 in form A.

[0331] Manufacturing details

[0332] Manufacturing details are provided in Table 16.

[0333] Table 16:

[0334] step# BOP# Starting quantity (Kg) Production amount (Kg) Yield (%) HPLC purity (%) 1 2493-1903-00484 27.5 kg 34.0kg 76.0％ 100.0％ 2 2479-1903-00489 28.8kg 48.6kg 96.4％ 99.1％ 3 2479-1904-00494 48.5kg 41.7kg 86.0％ 99.9％

[0335] Generating details

[0336] According to Scheme 2, step 1 is performed to form CV8814 free base (2493-1903-00487).

[0337] Option 2:

[0338]

[0339] Table 17 provides details on how step 1 was generated.

[0340] Table 17:

[0341]

[0342] 1) Sodium triacetoxyborohydride (STAB; 60.3 kg; CHP batch number: 181-190220) and 2-MeTHF (189.2 kg; CHP batch number: 234-190227) were loaded into reactor R-401.

[0343] 2) Stir the contents of R-401 and adjust the temperature to 15℃±5℃.

[0344] 3) 2,3,4-trimethoxybenzaldehyde (27.5 kg; CHP batch number: 275-190311, 142-190212) and 2-MeTHF (71.1 kg; CHP batch number: 234-190227) were loaded into reactor R-402.

[0345] 4) Start stirring reactor R-402.

[0346] 5) 2-(piperazin-1-yl)ethanol-1-ol (20.0 kg; CHP batch number: 173-190220) and 2-MeTHF (47.3 kg; CHP batch number: 234-190227) were loaded into reactor R-402.

[0347] 6) Stir the contents of reactor R-402 for at least 5 minutes.

[0348] 7) Add acetic acid (1.41 kg; CHP batch number: 151-190214) into reactor R-401 while keeping the temperature of the mixture below 25°C.

[0349] 8)T max =14.4℃.

[0350] 9) Transfer the contents of reactor R-402 over 1 hour and 19 minutes while keeping the temperature of the mixture below 25°C.

[0351] 10)T max =29.2℃ (temperature exceeded range during addition).

[0352] 11) The approved response continues.

[0353] 12) Adjust the temperature of the contents of R-401 to 20℃±5℃ and stir at 20℃±5℃ for at least 6 hours.

[0354] 13) Samples were taken from the contents of R-401 approximately 19 hours later.

[0355] 14) QC analysis of the sample indicated that no peak value of 2,3,4-trimethoxybenzaldehyde was detected. (Specification ≤ 1.5 area %)

[0356] 15) Adjust the temperature of the contents of R-401 to 15℃±5℃.

[0357] 16) Pour water (247.5 kg; CHP batch number: 232-190227) into R-401 while keeping the temperature below 25°C.

[0358] 17)T max =14.6℃.

[0359] 18) Adjust the temperature of the contents of R-401 to 20℃±5℃ and stir the contents for 30 minutes.

[0360] 19) Let the contents of R-401 stand for 30 minutes.

[0361] 20) Perform phase cutting and transfer the water layer to R-402.

[0362] 21) Load MTBE (203.8 kg; CHP batch number: 207-190222) into R-402.

[0363] 22) Adjust the temperature of the contents of R-402 to 0℃±5℃.

[0364] 23) Pack 50% NaOH (68.3 kg; CHP batch number: 145-190213, 150-190214) into R-402, while maintaining the temperature below 25℃.

[0365] 24)T max =11.5℃.

[0366] 25) After adding the contents, adjust the temperature of the contents of R-402 to 20℃±5℃.

[0367] 26) Stir the contents of R-402 for at least 30 minutes, then let the contents stand for at least 30 minutes.

[0368] 27) Perform phase cutting and transfer the water layer to reactor R-401, while retaining the organic layer in reactor R-402.

[0369] 28) Load MTBE (61.0 kg; CHP batch number: 207-190222) into R-401.

[0370] 29) Adjust the temperature of the contents of R-401 to 20℃±5℃.

[0371] 30) Stir the contents of R-401 for at least 15 minutes, then let the contents stand for at least 15 minutes.

[0372] 31) Perform phase cutting and transfer the water layer to the barrel, while retaining the organic layer in reactor R-401.

[0373] 32) Perform an FIO pH test on the bottled water layer, pH = 13.20.

[0374] 33) Transfer the contents of R-402 to R-401.

[0375] 34) Pour 20% NaCl solution (94.4 kg; NaCl: 19.0 kg, CHP batch number: 156-190214; water (75.9 kg; CHP batch number: 232-190227)) into R-401, stir for at least 15 minutes and let stand for at least 30 minutes.

[0376] 35) A performs phase cutting and transfers the water layer to the barrel.

[0377] 36) Distill the solution in R-401 under reduced pressure to a total volume of approximately 55 L, while maintaining the temperature at <45 °C.

[0378] 37) Load MTBE (61.1 kg; CHP batch number: 207-190222) into R-401.

[0379] 38) Distill the contents of R-401 under reduced pressure to a total volume of approximately 82 L, while maintaining the temperature at <45 °C.

[0380] 39) Load MTBE (61.1 kg; CHP batch number: 207-190222) into R-401.

[0381] 40) Distill the contents of R-101 under reduced pressure to a total volume of approximately 82 L, while maintaining the temperature at <45 °C.

[0382] 41) Sample the contents of R-401 (IPC sample: 2493-1903-00484-85-01) and check the water content of the solution by KF analysis.

[0383] 42) KF = 1.6% (specifications ≤ 0.5%)

[0384] 43) Load MTBE (61.1 kg; CHP batch number: 207-190222) into R-401.

[0385] 44) Distill the contents of R-101 under reduced pressure to a total volume of approximately 82 L, while maintaining the temperature at <45 °C.

[0386] 45) Sample the contents of R-401 (IPC sample: 2493-1903-00484-88-01) and check the water content of the solution by KF analysis.

[0387] 46) KF = 0.8% (specifications ≤ 0.5%)

[0388] 47) Load MTBE (61.1 kg; CHP batch number: 207-190222) into R-401.

[0389] 48) Distill the contents of R-101 under reduced pressure to a total volume of approximately 82 L, while maintaining the temperature at <45 °C.

[0390] 49) Sample the contents of R-401 (IPC sample: 2493-1903-00484-91-01) and check the water content of the solution by KF analysis.

[0391] 50) KF = 0.4% (specifications ≤ 0.5%)

[0392] 51) Load MTBE (30.9 kg; CHP batch number: 207-190222) into R-401.

[0393] 52) Adjust the temperature of the contents of R-401 to 40℃±5℃.

[0394] 53) Heptane (56.3 kg; CHP batch number: 233-190227) was loaded into R-401 over 8 minutes, while the temperature was maintained at 40℃±5℃.

[0395] 54)T min =38.1℃

[0396] 55) Obtain an FIO sample (FIO sample: 2493-1903-00484-100-01) to observe the MTBE:heptane ratio in R-401. The ratio was 1.5:6.0.

[0397] 56) Adjust the temperature of the contents of R-401 to 28°C ± 5°C (target 26°C to 29°C) for at least 30 minutes and stir at this temperature for at least 30 minutes.

[0398] 57) Solid formation was observed.

[0399] 58) Adjust the temperature of the contents of R-401 to 30℃±3℃ and stir for at least 30 minutes.

[0400] 59) Heptane (56.4 kg; CHP batch number: 233-190227) was loaded into R-401 over 24 minutes while maintaining the temperature at 30℃±5℃.

[0401] 60)T min =30.2℃.

[0402] 61) Adjust the temperature of the contents of R-401 to 30℃±3℃ and stir for at least 20 minutes.

[0403] 62) Adjust the temperature of the contents of R-401 to 20℃±5℃ for at least 30 minutes and stir for at least 20 minutes.

[0404] 63) Adjust the temperature of the contents of R-401 to 5℃±5℃ for at least 30 minutes and stir for at least 30 minutes.

[0405] 64) Collect solids on filter-FD-400.

[0406] 65) Wash the contents of filter-FD-400 with cold heptane (49.4 kg; CHP batch number: 233-190227).

[0407] 66) Dry the contents of filter-FD-400 under vacuum at ≤25°C with N2 gas flow for at least 16 hours.

[0408] 67) Submit the IPC sample (IPC sample: 2493-1903-00484-122-01) to QC to obtain LOD.

[0409] • LOD = 0.20% (specifications ≤ 0.5%)

[0410] 68) Pack twice the amount of the product CV-8814 free alkali (CV8814 free alkali) into a swan-neck bottle and weigh it.

[0411] 69) Analysis of dried material (IPC sample: 2493-1903-00484-122-01):

[0412] Appearance: White to off-white solid

[0413] • Weight: 34.0 kg (78.0% yield)

[0414] • HPLC purity = 100.0%

[0415] • 1H NMR: coincidence structure

[0416] 70) Take a 5kg portion of CV-8814 free alkali (CV8814 free alkali) from the twice-sized bagged and swan-neck bottled bulk material and reserve it for release according to batch number 2493-1903-00484.

[0417] Step 2 is performed according to scheme 3 to form CV8972 monohydrate (2479-1903-00489).

[0418] Option 3:

[0419]

[0420] Table 18 provides details on the generation of steps 2a and 2b.

[0421] Table 18:

[0422]

[0423]

[0424] 1) Add nicotinic acid (17.1 kg, CHP batch number 201-190222) and DCM (153.0 kg, CHP batch number 328-190326) into reactor R-401.

[0425] 2) Stir the contents of R-401 and adjust the temperature to 15±5℃.

[0426] 3) Load CV8814 free alkali (28.8 kg, CHP batch number 2493-1903-00484), EDC (26.7 kg, CHP batch number 147-190213), DMAP (1.70 kg, CHP batch number 152-190214) and DCM (306.6 kg, CHP batch number 328-190326) into reactor R-402.

[0427] 4) Stir the contents of R-402 for at least 20 minutes.

[0428] 5) Transfer the contents of R-402 to R-401 over a period of at least 30 minutes, while keeping the temperature below 25°C.

[0429] 6)T max =20.0℃.

[0430] 7) Adjust the temperature of the contents of R-401 to 20±5℃ and stir for at least 16 hours.

[0431] 8) Sample the contents of R-401 approximately 16 hours later.

[0432] 9) QC analysis of the sample indicated that 0% (0.05%) of CV-8814 free base (CV8814 free base) was detected relative to CV8972 (specification: ≤1% CV8814 free base).

[0433] 10) Adjust the contents of R-401 to 10±5℃.

[0434] 11) Slowly add water (29.2 kg, CHP batch number 329-190326) while keeping the temperature below 25°C.

[0435] 12)T max =12.3℃.

[0436] 13) Adjust the temperature of the contents of R-401 to 20±5℃, stir for at least 15 minutes and let stand for at least 15 minutes.

[0437] 14) Separated phase.

[0438] 15) Transfer the lower organic layer containing the product to reactor R-402. Send the water layer to the drum.

[0439] 16) Add water (29.0 kg, CHP batch number 329-190326) into the reactor.

[0440] 17) Stir the two-phase mixture for 15 minutes and let it stand for 15 minutes.

[0441] 18) Separated phase.

[0442] 19) Transfer the organic layer containing the product to reactor R-401. Send the water layer to the drum.

[0443] 20) Add 8% NaHCO3 aqueous solution (sodium bicarbonate, 4.0 kg, CHP batch number 192-190221; water, 53.2 kg, CHP batch number 329-190326) to the reactor.

[0444] 21) Stir the mixture for at least 15 minutes and let it stand for at least 15 minutes.

[0445] 22) Separated phase.

[0446] 23) Transfer the lower organic layer containing the product to reactor R-402. Send the water layer to the drum.

[0447] 24) Add water (29.0 kg, CHP batch number 329-190326) to the reactor.

[0448] 25) Stir the mixture for at least 15 minutes and let it stand for at least 15 minutes.

[0449] 26) R-401 was washed with water (17.6 kg, CHP batch number 329-190326) and MEK (5.9 kg, CHP batch number 330-190326) and dried with N2 airflow.

[0450] 27) Separated phase.

[0451] 28) Transfer the lower organic layer containing the product to reactor R-401. Send the water layer to the drum.

[0452] 29) Concentrate the contents of R-401 to approximately 72L under reduced pressure, while maintaining the temperature below 45°C.

[0453] 30)T max =32.0℃.

[0454] 31) Load MEK (139.0 kg, CHP batch number 330-190326) into R-401.

[0455] 32) Concentrate the contents of R-401 to approximately 72L under reduced pressure, while maintaining the temperature below 45°C.

[0456] 33)T max =32.0℃.

[0457] 34) Load MEK (139.1 kg, CHP batch number 330-190326) into R-401.

[0458] 35) Concentrate the contents of R-401 to approximately 72L under reduced pressure, while maintaining the temperature below 45°C.

[0459] 36)T max =29.2℃.

[0460] 37) FIO 1H NMR was used to determine the DCM:MEK ratio. DCM:MEK = 1:214.9

[0461] 38) Load MEK (185.5 kg, CHP batch number 330-190326) into R-401.

[0462] 39) Load MEK (208.7 kg, CHP batch number 330-190326) and concentrated HCl (30.2 kg, CHP batch number 274-190311) into a clean R-402 container.

[0463] 40) Adjust the temperature of the contents of the reactor to 25±5℃.

[0464] 41) Transfer the contents of R-401 to R-402 over approximately 1 hour, while maintaining the temperature below 35°C.

[0465] 42)T max =27.2℃.

[0466] 43) Heat the contents of R-402 to 50±5℃ and stir for at least 1 hour.

[0467] 44) Cool the contents of the reactor to 20±5℃ over 2 hours.

[0468] 45) Stir the contents of the reactor at 20±5℃ for 15 hours.

[0469] 46) Filter solids.

[0470] 47) Rinse the filter cake with MEK (58.0 kg, CHP batch number 330-190326).

[0471] 48) Rinse the filter cake with MEK (58.0 kg, CHP batch number 330-190326).

[0472] 49) Dry the wet filter cake in a disc dryer at 20-25°C for at least 16 hours, with no nitrogen leaching.

[0473] 50) Submit the IPC sample (IPC sample: 2479-1903-00489-87-01) to QC to obtain KF.

[0474] • Water content (specification: ≤4%, based on cKF): 3.3%

[0475] 51) Pack twice the product CV8972 monohydrate into bags, put them into swan-neck bottles, and weigh them.

[0476] 52) Analysis of dried material (IPC sample: 2479-1903-00489-87-01):

[0477] Appearance: White to off-white solid

[0478] • Weight: 48.6 kg (96.4% yield)

[0479] • 1H NMR: coincidence structure

[0480] • HPLC purity (area%) 99.1%

[0481] • GC analysis of residual solvent in FIO:

[0482] 2-MeTHF: No peak value

[0483] DCM: No peak value

[0484] MTBE: No peak value

[0485] Heptane: No peak value

[0486] MEK: 343ppm

[0487] Acetic acid: 874 ppm

[0488] According to scheme 4, step 3 is performed to form CV-8972 (2479-1904-00494).

[0489] Option 4:

[0490]

[0491] Table 19 provides details on how step 3 is generated.

[0492] Table 19:

[0493]

[0494] 1) CV8972 monohydrate (48.5 kg, CHP batch number 2479-1903-00489), water (48.5 kg, CHP batch number 329-190326), methanol (19.2 kg, CHP batch number 379-190404) and MEK (39.0 kg, CHP batch number 330-190326) were respectively loaded into reactor R-402.

[0495] 2) Adjust the contents of R-402 to 20℃±5℃ and stir until a solution is obtained.

[0496] 3) Transfer the contents of R-402 to R-401 through a 0.45-micron straight-through filter.

[0497] 4) Put water (24.0 kg, CHP batch number 329-190326) and methanol (19.2 kg, CHP batch number 379-190404) into R-402 and transfer them to R-401 through a 0.45 micron straight-through filter.

[0498] 5) Put methanol (192.1 kg, CHP batch number 379-190404) into R-401 through a 0.45 micron straight-through filter.

[0499] 6) Adjust the temperature of the contents of R-401 to 60±5℃.

[0500] 7) After about 2 hours, MEK (899.6 kg, CHP batch number 330-190326; 380-190404) is loaded into R-401 through a 0.45 micron straight-through filter, while the temperature is maintained at 60±5℃.

[0501] 8) Stir the contents of R-401 at 60±5℃ for at least 4 hours.

[0502] 9) Adjust the temperature of R-401 to 20±5℃ over at least 3 hours.

[0503] 10) Stir the contents of R-401 at 20±5℃ for about 9 hours.

[0504] 11) Send the contents of R-401 to the filter.

[0505] 12) Rinse the filter cake with MEK (105.4kg, CHP batch number 380-190404).

[0506] 13) Rinse the filter cake with MEK (105.4kg, CHP batch number 380-190404).

[0507] 14) Vacuum dry the wet filter cake on the filter for at least 30 minutes.

[0508] 15) Pack the wet filter cake in a filter dryer and dry it at ≤30°C under reduced pressure for at least 12 hours.

[0509] 16) Submit the IPC sample (IPC sample: 2479-1904-00494-32-01) to QC to obtain KF and residual solvent GC.

[0510] • Water content (specification: 2.8-3.8%, based on cKF) = 3.5%

[0511] • Residual solvent GC analysis (specifications: MeOH ≤ 3000 ppm; MEK ≤ 5000 ppm): MeOH = 215 ppm; MEK = 185 ppm

[0512] 17) Pack the product CV-8972 twice into bags, put it into swan-neck bottles, and weigh it.

[0513] 18) Analyze the dried material (IPC sample: 2479-1904-00494-32-01)

[0514] Appearance: White to off-white solid

[0515] • Weight = 41.7 kg (86.0% yield)

[0516] • HPLC purity (area%) = 99.9%

[0517] Known impurities:

[0518] Niacin: No peak value

[0519] DMAP: No peak value

[0520] CV8814: 0.1%

[0521] 2,3,4-Trimethoxybenzaldehyde; no peak value

[0522] Trimetazidine: No peak value

[0523] CV-10099: No peak value

[0524] CV-10046: No peak value

[0525] XRPD: Conformation A

[0526] Chloride ion content: 19.4%

[0527] • 1H NMR: coincidence structure

[0528] in conclusion

[0529] The results provided above indicate that CV-8972 can be synthesized using Scheme 1. In step 1, 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethanol-1-ol were used as starting materials, sodium triacetoxyborohydride (STAB) was used as a reducing agent, and acetic acid (AcOH) and 2-methyltetrahydrofuran (2-MeTHF) were used as catalysts for reductive amination. After aqueous post-treatment, solvent exchange, and crystallization, CV-8814 free base (CV8814 free base) with a purity of 100.0% and a yield of 78.0% was obtained by HPLC. A 5 kg sample of CV-8814 free base (CV8814 free base) was transferred from the synthesis for release. In step 2, CV8814 free base was coupled with nicotinic acid in DCM in the presence of EDC and catalytic DMAP to achieve the conversion of CV8972 free base by HPLC IPC. The solvent was exchanged to MEK and added to MEK containing concentrated HCl, yielding CV8972 monohydrate in 96.4% yield and 99.1% purity by HPLC. The final form conversion in step 3 was completed by heating the CV8972 monohydrate in a mixture of water, methanol, and MEK to 60℃ ± 5℃ and then adding MEK to precipitate it. A white solid CV-8972 in 86.0% yield and 99.9% purity was obtained by HPLC, and XRPD analysis confirmed it to be form A. The overall yield of the GMP synthesis of CV-8972 was 64.7%. The final amount of CV-8972 was 41.7 kg.

[0530] By incorporating via reference

[0531] Throughout this disclosure, other literature, such as patents, patent applications, patent publications, magazines, books, papers, and web content, has been referenced and cited. All such literature is hereby incorporated in its entirety for all purposes.

[0532] equivalent

[0533] Based on the entire contents of this document, including references to scientific patent literature cited herein, various modifications to the invention and many other embodiments thereof will become apparent to those skilled in the art, in addition to those shown and described herein. The subject matter of this document contains important information, illustrations, and guidance applicable to practicing the invention in its various embodiments and equivalents.

Claims

1. Form A crystalline of a compound of Formula (X): ###0001### Form A crystalline of a compound of Formula (X): ###0001### wherein the Form A crystalline has an XRPD pattern comprising peaks as shown in Figure 4. （X）， 2. The crystalline of claim 1, wherein the crystalline comprises a trihydrochloride salt of the compound.

3. The crystalline of claim 2, wherein the crystalline comprises a hydrated form of the compound, wherein the hydrated form of the compound is a monohydrate.

4. The crystalline of claim 1, wherein the compound has endothermic peaks in a differential scanning calorimetry (DSC) thermogram at 85.3 °C ± 5 °C and 214.6 °C ± 5 °C.

5. The crystalline of claim 4, the compound DSC thermogram is consistent with Figure 5.

6. The crystalline of claim 1, wherein the compound exhibits dehydration and a weight loss of 3.46% in a thermogravimetric analysis (TGA) at 25.9 °C to 150.0 °C.

7. The crystalline of claim 1, wherein the compound TGA thermogram is consistent with Figure 5.

8. A pharmaceutical composition comprising Form A crystalline of a compound of Formula (X): ###0001### Form A crystalline of a compound of Formula (X): ###0001### wherein the Form A crystalline has an XRPD pattern comprising peaks as shown in Figure 4.

9. The pharmaceutical composition of claim 8, wherein the composition comprises a trihydrochloride salt of the compound. （X）， 10. The pharmaceutical composition of claim 9, wherein the composition comprises a hydrated form of the compound, wherein the hydrated form of the compound is a monohydrate.

11. The pharmaceutical composition of claim 8, wherein the composition is formulated for oral administration.

12. The pharmaceutical composition of claim 8, wherein the composition is formulated as a single unit dosage.

13. The pharmaceutical composition of claim 8, wherein the composition is formulated as separate doses.

14. The pharmaceutical composition of claim 8, wherein the compound has endothermic peaks in a differential scanning calorimetry (DSC) thermogram at 85.3 °C ± 5 °C and 214.6 °C ± 5 °C.

15. The pharmaceutical composition of claim 14, the compound DSC thermogram is consistent with Figure 5.

16. The pharmaceutical composition of claim 8, wherein the compound exhibits dehydration and a weight loss of 3.46% in a thermogravimetric analysis (TGA) at 25.9 °C to 150.0 °C.

17. The pharmaceutical composition of claim 8, wherein the compound TGA thermogram is consistent with Figure 5.

18. Use of a composition comprising a therapeutically effective amount of Form A crystalline of a compound of Formula (X): ###0001### Form A crystalline of a compound of Formula (X): ###0001### wherein the Form A crystalline has an XRPD pattern comprising peaks as shown in Figure 4, in the manufacture of a medicament for the treatment and / or prevention of a condition.

19. The use of claim 18, wherein the composition comprises a trihydrochloride salt of the compound.

20. The use of claim 19, wherein the composition comprises a hydrated form of the compound, wherein the hydrated form of the compound is a monohydrate. （X）， ​ ​ ​ 21. The use of claim 18, wherein the composition is provided orally.

22. The use of claim 18, wherein the composition is provided as a single unit dose.

23. The use of claim 18, wherein the condition is selected from the group consisting of an aneurysm, cerebrovascular disease, heart disease, and hypertension.

24. The use of claim 18, wherein the condition is selected from the group consisting of cardiomyopathy, congenital heart disease, ischemic heart disease, pericardial disease, rheumatic heart disease onset, heart failure, and valvular heart disease.

25. The use of claim 18, wherein the condition is diabetic cardiomyopathy.

26. The use of claim 18, wherein the condition is atherosclerosis.

27. The use of claim 18, wherein the condition is stroke or transient ischemic attack.

28. The use of claim 18, wherein the condition is angina pectoris.

29. The use of claim 18, wherein the condition is peripheral arterial disease.

30. The use of claim 18, wherein the compound has endothermic peaks in differential scanning calorimetry (DSC) thermogram at 85.3 °C ± 5 °C and 214.6 °C ± 5 °C.

31. The use of claim 18, wherein the compound DSC thermogram is consistent with FIG.

5.

32. The use of claim 18, wherein the compound exhibits dehydration in thermogravimetric analysis (TGA) at 25.9 °C to 150.0 °C and a weight loss of 3.46%.

33. The use of claim 18, wherein the compound TGA thermogram is consistent with FIG. 5.

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